Capa Livro 12 Simp Int Esquis... - Pide

Transcrição

SUMÁRIO
(Contents)
Comissão Organizadora ................................................................................ 5
(Organizing Committee)
Mensagem de Boas Vindas .......................................................................... 7
(Welcome Message)
Simpósios Realizados.................................................................................... 9
(Symposia been held)
Estrutura do Programa ............................................................................... 10
(Programme Structure)
Resumo das Atividades Científicas ............................................................. 11
(Abstracts of Scientific Ativities)
Programação Científica............................................................................... 14
(Cientific Programme)
Programação Temas Livres ......................................................................... 23
(Free Theme Programme)
Programação Posters .................................................................................. 29
(Poster Programme)
Resumos de Mesas Redondas .................................................................... 51
(Table Abstracts Round)
Resumos de Temas Livres e Posters ......................................................... 103
(Free Theme and Posters Abstracts)
Índice de Autores ..................................................................................... 415
(Author Index)
International Symposium on Schistosomiasis
Eridan de Medeiros Coutinho
Presidente de Honra
(President of Honour)
COMISSãO ORganIzadORa
(Organizing Committee)
Carlos Eduardo Grault
(President)
Otávio Sarmento Pieri
Tereza Cristina Favre
Silvana Thiengo
Omar Carvalho
Liana Konovaloff Jannotti Passos
Comissão Científica
(Scientific Committee)
Comissão Executiva
(Executive Committe)
Ana Lúcia Coutinho Domingues
Carlos Graeff Teixeira
Constança Simões Barbosa
Cristiano Lara Massara
Guilherme Correa Oliveira
Henrique Leonel Lenzi
José Roberto Machado e Silva
Martin Johannes Enk
Mitermayer Galvão dos Reis
Naftale Kat
Paulo Marcos Zech Coelho
Roberta Lima Caldeira
Rodrigo Correa de Oliveira
(Coordinator)
Stefan Greiger)
Virgínia Schall
Zilton Andrade
Aline Carvalho Mattos
Aline Favre Galvão
Lilian Beck
Monica Ammon
Pablo Menezes Coelho
Renata Savignon Rubim
Capa (Cover): Projeto Gráfico (Graphic Project): Ruben Fernandes
Foto (Photo): Vinicius Marinho (Fiocruz/Multi Imagens)
5
WELCOME MESSagE
Dear Colleagues,
We are very pleased to welcome you to the 12th International
Symposium on Schistosomiasis, organized by the Integrated Schistosomiasis Programme of the Oswaldo Cruz Foundation (PIDE).
The theme of the Symposium will be new knowledge on schistosomiasis based on a broad approach of research and technological
development, including interprogram strategies, intersectorial actions and socioenvironmental determinants aimed at equity in health
access and effectivity in prevention, surveillance and control. The
topics range from recent advances in genomics, immunopathology,
biochemistry, molecular biology and ecoepidemiology to innovations
in diagnosis, treatment and health education, to be presented in conferences, round tables and courses as well as free theme and poster
sessions.
More than 400 participants, including 58 invited speakers from
various national and international institutions, will be present to discuss 253 scientific works. This will be an excellent opportunity for
researchers from different areas of knowledge to join together with
health managers and policy makers for an integrated, multidisciplinary discussion on challenges and achievements in the combat of this
endemic disease, thus contributing to promote the incorporation of
scientific knowledge into the control programmes.
We are sure that, by the end of the Symposium, all participants
will feel to be closer to the aim of reducing this important povertyrelated disease to a level so that it will not be considered as a health
problem in the near future.
Carlos Eduardo Grault
Symposium President
Omar Carvalho
PIDE Coordinator
7
SIMPóSIOS REaLIzadOS
(Symposia been held)
9
EStRUtURa dO PROgRaMa
(Programme Structure)
10
RESUMO daS atIvIdadES CIEntífICaS
(Summary of Scientific Ativities)
11
12
13
PROgRaMaçãO CIEntífICa (Cientific Programme)
14
15
16
17
18
19
20
21
22
PROgRaMaçãO dE tEMaS LIvRES
(Free Theme Programme)
06/10 (14h00 - 15h30)
Sala (Room) guaratiba
Epidemiologia, Controle e Educação em Saúde
(Epidemiology, Control and Health Education)
Coordenador (Chair): Naftale Katz (CPqRR, Fiocruz/MG)
01. EVALUATION OF THE IMPLEMENTATION SCHISTOSOMIASIS MANSONI EPIDEMIOLOGICAL
SURVEILLANCE: A STUDY CASE IN THE MUNICIPALITY OF UNIÃO DOS PALMARES IN
ALAGOAS STATE
Jeann Marie Rocha Marcelino; Luciano Medeiros de Toledo; Helia Kawa; Marly Marques
da Cruz
02. PATTERN AND IMPACT OH HELMINTH INFECTIONS IN PREGNANCY: PROSPECTS FOR
IMPROVEMENT MATERNAL HEALTH IN ANGOLA
Silvana Maria Duarte Belo; Ilda Rosalina Jeremias; Silvana Maria Duarte Belo; Joltim
Quivinja; Filomena Gomes da Silva; Eugénia Ramos; Maria Amélia Afonso Grácio
03. SURVEY OF SOCIO EPIDEMIOLOGICAL DATA ON PATIENTS WITH SCHISTOSOMIASIS OF
CAPELA DO SOCORRO, SOUTH REGION OF SÃO PAULO, IN THE PERIOD OF 2004 TO 2010
Beatriz Aparecida Imparato; Andréia Aparecida Caggegi; Rafael de Oliveira Christe;
Nathália Rafaella Martinho; Elaine Cristina S Gama; Léia Munhoz Parra; Ana Lúcia de
Lima Gabriel; Patricia Placoná Diniz
04. CONSTRUTION OF COMPUTERIZED TEMPLATES FOR EPIDEMIOLOGICAL SURVEYS ON
SCHISTOSOMIASIS: EXPERIENCE IN PORTO DE GALINHAS, PERNAMBUCO
Constança Simões Barbosa; Elainne Christine Souza Gomes; Onicio Batista Leal Neto
05. EVALUATION OF PARASITOLOGICAL DIAGNOSTIC METHODS FOR SCHISTOSOMIASIS
MANSONI IN LOW TRANSMISION ÁREA IN MINAS GERAIS, BRAZIL
Liliane Maria Vidal Siqueira; Áureo Almeida de Oliveira; Cristiano Lara Massara; Nidia
Francisca de Figueiredo Carneiro; Paulo Marcos Zech Coelho; Martin Johannes Enk
06. SOCIOENVIRONMENTAL PROFILE OF PATIENTS WITH SCHISTOSOMIASIS MANSONI IN
LOW ENDEMIC SETTING IN CEARA STATE
Fernando Schemelzer de Moraes Bezerra; Marta Cristhiany Cunha Pinheiro; Mariana
Silva Sousa; Teiliane Rodrigues Carneiro; Sara Menezes de Oliveira; Maria de Fátima
Oliveira
07. THE APPLICATION OF GEOGRAPHIC INFORMATION SYSTEM (GIS) DURING A ONE YEAR
OBSERVATION PERIOD OF INFECTIONS WITH SCHISTOSOMA MANSONI IN PEDRA PRETA,
MUNICIPALITY OF MONTES CLAROS, MINAS GERAIS – BRAZIL
Ricardo José de Paula Souza e Guimarães; Cristiano Lara Massara; Martin Johannes Enk
23
06/10 (14h00 - 15h30)
Sala (Room) São Conrado
Imunopatologia, (Immunopathology)
Coordenador (Chair): Silvia Montenegro (CPqAM, Fiocruz/PE)
08. ULTRASTRUCTURAL MORPHOLOGY OF ADULT SCHISTOSOMA MANSONI, HARBORED
IN NON ANTI-HELMINTHIC TREATED, HIGH-IMMUNE-TOLEROGENIC AND LOWINFLAMMATORY MICE BY TRANSMISSION ELECTRON MICROSCOPY
Aurelizia Maria Lemos Xavier; Antonio Carlos Silva; Daniel Tavares; Beatriz Ferreira
Ribeiro; Susane Borges Rodrigues; Erick Vaz Guimarães; Luis Cláudio Muniz-Pereira;
Maria de Fátima Sarro-Silva; Antonio Henrique Almeida de Moraes Neto
09. CPG-ODN IS AN ALTERNATIVE ADJUVANT TO BE USED IN A VACCINE FORMULATION
AGAINST SCHISTOSOMIASIS
Juliano Michel De Araújo; Tatiane Teixeira de Melo; Ludmila Zanandreis de Mendonça;
Paulo Marcos Zech Coelho; Cristina Toscano Fonseca
10. EVALUATION OF THE N-ACETYLCYSTEINE IN THE IMMUNOPATHOLOGY OF THE
EXPERIMENTAL SCHISTOSOMIASIS MANSONI
André de Lima Aires; Renata Alexandre Ramos Silva; Tiago Moreira Alves Feitosa; Luiz
Henrique de Souza Teixeira; Giuliana Viegas Schirato; Nicodemos Teles de Pontes Filho;
Sidcley Bernardino de Araújo; Valdênia Maria Oliveira Souza; Vlaudia Maria Assis Costa;
Elizabeth Malagueño de Santana; Mônica Camelo Pessoa de Azevedo Albuquerque
11. THE EFFECT OF SCHISTOSOMA MANSONI ANTIGENS IN DOWN-MODULATE THE
INFLAMMATORY RESPONSE IN TH2-MEDIATED DISEASE
Luciana Santos Cardoso; Sérgio Costa Oliveira; Alfredo Miranda Góes; Ricardo Riccio
Oliveira; Robson da Paixão de Souza; Edgar M Carvalho; Maria Ilma Araujo
12. IMUNODOMINANCE OF THE SM-P40 ANTIGEN CONTRIBUTES TO THE DEVELOPMENT OF
SEVERE SCHISTOSOMIASIS MANSONI
Eduardo Finger; Thaissa Melo Galante Coimbra; Daniel Maurício dos Santos
13. RECOGNITION OF SCHISTOSOME TEGUMENT PROTEINS BY SERA FROM PUTATIVE
RESISTANT AND CHRONICALLY INFECTED HUMANS USING A PROTEIN MICROARRAY
Soraya Gaze; Patrick Driguez; Angela Trieu; Jeffrey Bethony; Fernanda Cardoso; Rodrigo
Correa-Oliveira; Phil Felgner; Donald McManus; Denise Doolan; Alex Loukas
14. THE USE OF SCHISTOSOMA MANSONI ANTIGENS TO PREVENT THE INFLAMMATORY
RESPONSE IN VITRO IN CUTANEOUS LEISHMANIASIS
Aline Michelle Barbosa Bafica; Giuseppe Tittone Varella; Ricardo Riccio Oliveira; Luciana
Santos Cardoso; Alfredo Góes; Sérgio Costa Oliveira; Alex Loukas; Edgar Marcelino
Carvalho; Maria Ilma Araújo
24
06/10 (14h00 - 15h30)
Sala (Room) Pontal
Biologia Molecular e Bioquímica (Molecular Biology and Biochemistry)
Coordenador (Chair): Ricardo DeMarco (USP/SP)
15. DIFFERENTIAL PROTEIN PARTITIONING DURING TEGUMENT ISOLATION OF THE PARASITE
SCHISTOSOMA MANSONI
Priscila Tavares Meneses; Sonaly Cristine Leal; Tiago Ferreira Leal; Milton Hercules Guerra
de Andrade; R. Alan Wilson; William de Castro Borges
16. THE SCHISTOSOMA MANSONI PHYLOME: EVOLUTIONARY HISTORIES TO IMPROVE
FUNCTIONAL PREDICTIONS AND GET INSIGHTS INTO PARASITE BIOLOGY
Larissa Lopes Silva; Marina Marcet-Houben; Laila Alves Nahum; Adhemar Zerlotini Neto;
Toni Gabaldón; Guilherme Oliveira
17. TOWARDS AN UNDERSTANDING OF THE EPIGENETICS OF SCHISTOSOMES
Céline Cosseau; Julie Lepesant; Jérome Boissier; Julien Portella; Cécile Perrin; Christoph
Grunau
18. NUCLEAR EXPORT AND SECRETION OF SCHISTOSOMA MANSONI HMGB1 PROTEIN IS
MEDIATED BY PHOSPHORYLATION
Isabel Caetano de Abreu da Silva; Vitor Coutinho Carneiro; Renata de Moraes Maciel;
Rodrigo Furtado Madeiro da Costa; Daniel Rodrigues Furtado; Francisco Meireles; Mario
Alberto Cardoso da Silva-Neto; Franklin David Rumjanek; Marcelo Rosado Fantappié
19. NUCLEOCYTOPLASMIC SHUTTLING OF SCHISTOSOMA MANSONI HMGB1 IS REGULATED
BY ACETYLATION THAT DIRECTS IT TOWARD SECRETION
Vitor Coutinho Carneiro; Isabel Caetano de Abreu da Silva; Claudia Neto Paiva; Renata
de Moraes Maciel dos Santos; Flavia Lamarão; Marcelo Pelajo Machado; Hélio Dutra;
Marcelo Torres Bozza; Marcelo Rosado Fantappie
20. IN SEARCH OF A PHARMACOPHORIC MODEL FOR SCHISTOSOMICIDAL BENZODIAZEPINES
Jean Pierre Barros Thibaut; Carla Maria Souza Menezes; Gildardo Rivera; Nathália Couto
Dias; Marina Amaral Alves; Eliezer J. Barreiro; Lídia Moreira Lima; François Noel
21. DLCS AS NEW VACCINE CANDIDATES AGAINST SCHISTOSOMIASIS
Patricia Placoná Diniz; Erika Nakajima; Patricia Aoki Miyasato; Toshie Kawano; Elizabeth
Angelica Leme Martins
25
07/10 (14h00 - 15h30)
Sala (Room) guaratiba
Hospedeiros Intermediários (Intermediate hosts)
Coordenador (Chair): Roberta Lima Caldeira (CPqRR, Fiocruz/MG)
22. EVALUATION OF THE INTERACTION BETWEEN PRIMARY AND SECONDARY SPOROCYSTS
OF SCHISTOSOMA MANSONI EXPOSED TO THE INTERNAL DEFENSE SYSTEM OF
BIOMPHALARIA TENAGOPHILA AND SUSCEPTIBLE TO THE PARASITE
Ana Carolina Alves de Mattos; Raquel Lopes Martins-Souza; Paulo Marcos Zech Coelho
23. INVESTIGATION OF HYBRIDISM BETWEEN BIOMPHALARIA COUSINI AND B. AMAZONICA
Tatiana Maria Teodoro; Liana Konovaloff Jannotti-Passos; Omar dos Santos Carvalho;
24. CONTROVERSIES REGARDING THE SIGNIFICANCE OF THE SO-CALLED “APO” “AMEBOCYTE
PRODUCING ORGAN” IN BIOMPHALARIA GLABRATA
Samaly dos Santos Souza; Zilton de Araújo Andrade;
25. MALACOLOGICAL SURVEY OF BIOMPHALARIA IN THE MUNICIPALITIES OF “ESTRADA
REAL” IN SOUTHWESTERN OF MINAS GERAIS STATE, BRAZIL
Sandra Helena Cerrato Tibiriçá; Adalberto Mittherofhe; Milton F. Castro; Adilson C. Lima;
Murilo Gonçalves; Isabella O. Pinheiro; Corina C. Freitas; Ricardo J.P.S Guimarães; Omar
Santos Carvalho; Elaine Soares Coimbra
26. RISK AREAS OF SCHISTOSOMIASIS NEAR THE BRAZILIAN BORDER INTO THE URUGUAY
RIVERS BASINS IN THE PROVINCE OF CORRIENTES, ARGENTINA
Maria JF Rea; C. Edgardo Borda; Osvaldo D. Benitez; Luis A. Mosqueda
27. PARASITIC CASTRATION IN BIOMPHALARIA GLABRATA DURING THE PRE-PATENT
INFECTION WITH SCHISTOSOMA MANSONI
Marta Julia Faro dos Santos Costa; Mariana Perazzini; Lygia dos Reis Corrêa; Clélia
Christina Mello Silva; Arnaldo Maldonado Junior
28. MAINTENANCE OF HUMAN AND WILD CYCLE OF SCHISTOSOMA MANSONI IN
LABORATORY
Marjane Soares Ferreira; Andiara Garcez de Souza Silva; Selma Patrícia Diniz Cantanhede;
Luciana Patrícia Lima Alves; Hallyne Davinck Mesquita Moreira; Nêuton Silva-Souza;
Juliana Araripe Gomes da Silva
26
07/10 (14h00 - 15h30)
Sala (Room) São Conrado
Diagnóstico, Tratamento e Clínica (Diagnosis, Treatment and Clinical)
Coordenador (Chair): Cristina Toscano (CPqRR, Fiocruz/MG)
29. EVALUATION OF THE EFFECT OF THE ASSOCIATION IMMUNIZATION-CHEMOTHERAPY ON
SCHISTOSOMA MANSONI INFECTION
Ludmila Zanandreis de Mendonça; Patricia Martins Parreiras; Tatiane Teixeira de Melo;
Neusa Araújo Pereira; Juliano Michel Araújo; Flávia Fernanda Búbula Couto; Ana Carolina
Alves de Mattos; Paulo Marcos Zech Coelho; Cristina Toscano Fonseca
30. EVALUATION OF THE TAQMAN REAL TIME PCR AS AN ALTERNATIVE FOR DETECTION
OF SCHISTOSOMA MANSONI EGGS IN FECES AFTER THE CONCENTRATION HELMINTEX
METHOD
Candida Fagundes Teixeira; Aline Cardoso Caseca Volotão; Carlos Graeff Teixeira
31. MOLECULAR ANALYSIS OF THE OVIPOSITORY BEHAVIOR OF MURINE SCHISTOSOMIASIS
Eduardo Finger; Thaissa Melo Galante Coimbra; Érika Lopes Fernandes; Débora Cristina
Rosseto Garotti
32. 13947 IDENTIFICATION OF SCHISTOSOMA MANSONI ANTIGENS CANDIDATES TO BE USED
IN THE DIAGNOSIS OF SCHISTOSOMIASIS
Gardênia Braz Figueiredo de Carvalho; Cintia Maria Gonçalves da Silva; Lucila Gonçalves
Grossi Pacífico; Cristina Toscano Fonseca
33. EVALUATION OF HEPATIC FIBROSIS IN SCHISTOSOMIASIS BY BIOLOGICAL SERUM
MARKERS
Tibério Batista de Medeiros; Ana Lúcia Coutinho Domingues; Edmundo Pessoa de
Almeida Lopes; Ana Virgínia Matos Sá Barreto; Clarice Neuenschwander Lins de Morais;
Silvia Maria Lucena Montenegro; José Roberto Maciel Martins
34. AN OUTBREAK OF ACUTE SCHISTOSOMIASIS IN THE SOUTHEAST OF BRAZIL: II. CLINICAL
ASPECTS
José Roberto Lambertucci; Sandra Costa Drummond; Izabela Voieta; Alba Otoni; Bruna
Assis Chaves; Pedro Henrique Prata; Pedro Paulo Nunes Pereira; Thiago Cardoso Vale;
Leonardo Campos de Queiroz; Paloma Fonseca; Carlos Maurício de Figueiredo Antunes
35. PROLONGED COURSE OF SCHISTOSOMAL PULMONARY HYPERTENSION
Rita de Cassia dos Santos Ferreira; Ana Lúcia Coutinho Domingues; Angela Pontes
Bandeira; Amanda Medeiros Gomes da Silva
27
07/10 (14h00 - 15h30)
Sala (Room) Pontal
Epidemiologia, Controle e Educação em Saúde
(Epidemiology, Control and Health Education)
Coordenador (Chair): Martin Johannes Enk (CPqRR/MG)
36. HEALTH SURVEILLANCE AND BASIC HEALTH ATENTION, AN INTEGRATION EXEMPLE ON
THE SCHISTOSOMIASIS CONTROL IN JABOATÃO DOS GUARARAPES, PERNAMBUCO,
BRASIL - 2010
Francine Nesello; Tânia Gomes de Carvalho; Ana Carolina Cintra M. Medeiros; Rodriga
Maria Zovka de Souza; Sérgio Carneiro Vieira da Rocha; José Alexandre Menezes da Silva
37. AN AUTOMATIC PROPOSAL FOR DIAGNOSIS OF SCHISTOSOMIASIS
André Caetano Alves Firmo; Carmelo j. A. Bastos Filho; Jones Albuquerque; Silvana
Bocanegra; Reinaldo Souza Santos; Constança S. Barbosa
38. RISK FACTORS FOR SCHISTOSOMIASIS IN A LOW ENDEMIC URBAN ÁREA IN BRAZIL
Sandra Helena Cerrato Tibiriçá; Elaine Soares Coimbra; Clarice Abramo; Adalberto
Mittherofhe; Milton F. Castro; Isabella O. Pinheiro; Luiz Claudio Ribeiro; Maria Teresa
Bustamante Teixeira
39. EVALUATION OF SCHISTOSOMIASIS MORBIDITY BY CLINICAL AND ULTRASOUND
EXAMINATION, AND GEOREFERENCING MAPING THE SEVERE CASE IN THE ENDEMIC
ÁREA OF ILHA DAS FLORES, SERGIPE, BRAZIL
Karla Carolline Vieira Rollemberg; Carla Virginia Vieira Rollemberg; Bruno Leonardo
Nascimento Fernandes; Anny Caroline Porto Chagas; Fábio Ramalho de Amorim; Marília
M. B. L. Silva; Ângela Maria da Silva; Mário Adriano dos Santos; José Antônio P. de
Almeida; Amélia Maria Ribeiro de Jesus
40. IMPORTANCE OF PARTNERSHIP BETWEEN THE ENVIRONMENTAL MONITORING
PROGRAM AND FAMILY HEALTH IN THE CONTROL OF SCHISTOSOMIASIS IN JABOATÃO
DOS GUARARAPES, PERNAMBUCO, BRAZIL
Tânia Gomes de Carvalho; Vitor Alexandre Kessler de Almeida; José Alexandre Menezes
da Silva; Rodriga Maria Zovka de Souza; Sérgio Carneiro Vieira da Rocha; Francine Nesello
41. A COMMUNITY-BASED PROGRAM: HEALTH ACCESS AND SCHISTOSOMIASIS PREVENTION
AND CONTROL IN JEQUITINHONHA MUNCICIPALITY, MINAS GERAIS STATE, BRAZIL
Dener Carlos dos Reis; Maria Flávia Gazzinelli; Cristiano Lara Massara; Luiza Valgas de
Paula; Helmut Kloos; Rodrigo Corrêa Oliveira; Andréa Gazzinelli
42. FROM PREVENTION TO HEALTH PROMOTION: A STUDY WITH HEALTH AGENTS OF A
MUNICIPAL SCHISTOSOMIASIS PROGRAM
Danielle Grynszpan; Luciano Mendonça; Fernanda Sandes Cardoso
28
PROgRaMaçãO POStERS
(Posters Programme)
06/10 (18h00-19h00)
LOCaL: fOYER
Epidemiologia e Controle, Educação em Saúde,
Hospedeiros Intermediários, diagnóstico e tratamento
(Epidemiology, Control, Health Education,
Intermediate Hosts, Diagnosis and Treatment)
01. EVALUATION OF THE INSERTION OF GENETIC PATRIMONY OF BIOMPHALARIA
TENAGOPHILA TAIM (RESISTANT TO SCHSITOSOMA MANSONI) IN SNAILS COLLECTED IN
SCHISTOSOMIASIS ENDEMIC AREAS IN BANANAL/SP, BRAZIL
Daisymara Priscila de Almeida Marques; Florence Mara Rosa; Deborah Aparecida
Negrão-Corrêa3; Horácio Manuel Santana Teles; Engels Maciel; Roberta Lima Caldeira;
Áureo Almeida Oliveira; Paulo Marcos Zech Coelho
02. ANALISYS OF THE IN VITRO POTENCIAL SCHISTOSOMICIDAL OF THE CANDEIA EXTRACTS
Priscila Silva Grijó Farani; Laura Ambrósio Tosta; Luisa Maria Silveira de Almeida; Marcelo
Silva Silvério; Ana Carolina Alves de Mattos; Orlando Vieira de Sousa; Paulo Marcos Zech
Coelho; Eveline Gomes Vasconcelos; Priscila de Faria Pinto
03. ANALYSIS OF ANTAGONISTIC DRUGS TO CALCIUM CHANNELS IN SCHISTOSOMA MANSONI
ADULT WORMS
Flávia Fernanda Búbula Couto; Marah Mileib Vasconcelos; Neusa Araújo; Paulo Marcos
Zech Coelho; Rafaella Fortini Grenfell e Queiroz
04. ANGIOGENESIS: A QUANTITATIVE EVALUATION IN THE LIVER OF MICE WITH
SCHISTOSOMIASIS, BEFORE AND AFTER SPECIFIC CHEMOTHERAPY
Elisângela Trindade Santos; Márcia Maria de Souza; Ana Cristina Gonzalez; Zilton de
Araújo Andrade
05. ANTISCHISTOSOMAL EVALUATION OF THE IMIDAZOLINE DERIVATIVE 3-(4-CHLOROBENZYL)-5(4-NITRO-BENZYLIDINE) IMIDAZOLIDINE-2,4-DIONE
Marek Henryque Ferreira Ekert; Juliana Kelle de Andrade Lemoine Neves; André de Lima
Aires; Tiago Moreira Alves Feitosa; Renata Alexandre Ramos da Silva; Maria do Carmo
Alves de Lima; Ivan da Rocha Pitta; Suely Lins Galdino; Mônica Camelo Pessoa de Azevedo
Albuquerque
06. ASSESSMENT OF LIVER FIBROSIS IN SCHISTOSOMIASIS BY BIOCHEMICAL MARKERS AND
PLATELET COUNT
Vinícius Martins Alecrim; Ana Virgínia Matos Sá Barreto; Tibério B. Medeiros; Edmundo P.
A. Lopes; Ana Lúcia Coutinho Domingues; Clarice Neuenschwander Lins de Morais; Sílvia
Maria Lucena Montenegro
29
07. AVALIAR A OCORRENCIA DE VIRUS DE HEPATITES B E C EM PORTADORES DE
ESQUISTOSSOMOSE MANSONICA EM RESIDENTES EM AREA ENDEMICA, ALAGOAS
Maria Sonia Correia Alves; Danielle Correia Gama; Luciano Fernandes Pereira; Andrei
Leite Gazzaneo; Roberta Maria Pereira Albuquerque de Melo; Hugo Cabral Tenório;
Rozangela Maria de Almeida Fernandes Wyszomirska
08. BCG EXPRESSING SM-STOLP-2, A TEGUMENTAL PROTEIN OF SCHISTOSOMA MANSONI,
FAILS TO PROTECT MICE AGAINST CERCARIAL CHALLENGE
Alex Issamu Kanno; Leonardo Paiva Farias; Henrique Krambeck Rofatto; Cibele Aparecida
Tararam; Bogar Omar Araujo Montoya; Patrícia Aoki Miyasato; Eliana Nakano; Luciana
Cezar de Cerqueira Leite
09. BIOLOGICAL ACTIVITY OF SIDA PILOSA PLANT EXTRACT ON SCHISTOSOMA MANSONI
Hermine B.Jatsa; Cíntia Ap. de Jesus Pereira; Emília S. Araújo; Jaílza Lima Rodrigues;
Vinícius Gustavo Oliveira; Vanessa Fernandes Rodrigues; Florence M. Rosa; Fernão Castro
Braga; Mauro Martins Teixeira; Deborah Negrão-Corrêa
10. COMPARISON OF A POLYMERASE CHAIN REACTION AND THE KATO-KATZ TECHNIQUE
FOR THE DIAGNOSOSIS OF SCHISTOSOMA MANSONI IN A LOW ENDEMICITY AREA OF
MINAS GERAIS STATE, BRAZIL
Gabriel Costa de Carvalho; Letícia Helena dos Santos Marques; Luciana Inácia Gomes;
Ana Rabello; Luiz Cláudio Ribeiro; Sandra Helena Cerrato Tibiriçá; Elaine Soares Coimbra;
Clarice Abramo
11. CONVENTIONAL PCR FOR HUMAN SCHISTOSOMIASIS DIAGNOSIS IN STOOL SAMPLES
FROM INDIVIDUALS WITH LOW ELIMINATION OF PARASITE EGGS
Teiliane Rodrigues Carneiro; Regina Helena Saramago Peralta; Bárbara de Araújo Lima
Dutra; Marta Cristhiany Cunha Pinheiro; Sara Menezes de Oliveira; Fernando Schemelzer
de Moraes Bezerra; José Mauro Peralta
12. CURCUMIN-LOADED PLGA NANOPARTICLES: PREPARATION AND IN VITRO
SCHISTOSOMICIDAL ACTIVITY.
Priscilla Paiva Luz; Lizandra G. Magalhães; Wilson R. Cunha; Kátia Jorge Ciuffi; Eduardo
J. Nassar; Paulo Sergio Calefi; Emerson H. de Faria; Jairo K. Bastos; Vanderlei Rodrigues;
Márcio L. A. e Silva
13. DIFFERENTIAL REACTIVITY OF IGG1 AND IGG4 SUBTYPES FROM THE SCHISTOSOMIASIS
PATIENTS AGAINST SYNTHETIC PEPTIDES, BEFORE AND AFTER CHEMOTHERAPY
Karen Takaki Flores; Gabriane Nascimento Porcino; Rita Gabriela Pedrosa Ribeiro
Mendes; Michélia Antônia do Nascimento Gusmão; Cristiane de Carvalho Campos; Ana
Carolina Ribeiro Gomes Maia; Michelle de Lima Detoni; Danielle Gomes Marconato;
Maria Aparecida Juliano; Luiz Juliano; Paulo Marcos Zech Coelho; Priscila de Faria Pinto;
Eveline Gomes Vasconcelos
14. EFFECT OF DEXAMETHASONE IN THE TREATMENT FOR EXPERIMENTAL SCHISTOSOMIASIS
MANSONI
Neusa Araújo; Paulo Marcos Zech Coelho; Naftale Katz
30
15. EFFECT OF GARCINIA BRASILIENSIS IN INFECTION BY SCHISTOSOMA MANSONI
Liliane Gonçalves Vila Nova; Amanda Esteves Rocha; Marcos José Marques; Marcelo
Henrique dos Santos; Raquel Lopes Martins Souza
16. EVALUATION OF COPROSCOPIC METHODS (KATO-KATZ AND HELMINTEX) FOR DIAGNOSIS
OF SCHISTOSOMIASIS MANSONI IN LOW ENDEMIC SETTING IN CEARÁ STATE – BRAZIL
Marta Cristhiany Cunha Pinheiro; Teiliane Rodrigues Carneiro; Thaís Helena Guilherme
de Almeida; Sara Menezes de Oliveira; Mônica Coelho Andrade; Fernando Schemelzer de
Moraes Bezerra
17. EVALUATION OF COPROSCOPIC METHODS (KATO-KATZ AND SALT GRADIENT) FOR
DIAGNOSIS OF SCHISTOSOMIASIS MANSONI IN LOW ENDEMIC SETTING IN CEARÁ STATE
– BRAZIL
Marta Cristhiany Cunha Pinheiro; Ana Lúcia de Paula Hanemann; Teiliane Rodrigues
Carneiro; José Ajax Nogueira Queiroz; Aparecida Tiemi Nagao Dias; Sara Menezes de
Oliveira; Fernando Schemelzer de Moraes Bezerra
18. EVALUATION OF COPROSCOPIC METHODS (SALT GRADIENT AND HELMINTEX) FOR
DIAGNOSIS OF SCHISTOSOMIASIS MANSONI IN LOW ENDEMIC SETTING IN CEARÁ STATE
– BRAZIL
Marta Cristhiany Cunha Pinheiro; Teiliane Rodrigues Carneiro; Sara Menezes de Oliveira;
Ana Lúcia de Paula Hanemann; Fernando Schemelzer de Moraes Bezerra
19. EVALUATION OF EFFECT OF PRAZIQUANTEL ON ECHINOSTOMA PARAENSEI
(ECHINOSTOMATIDAE: PLATYHELMINTHES)
Júlia Peralta Gonçalves; Aleksandra Menezes de Oliveira; Arnaldo Maldonado Júnior;
Técia Maria Ulisses de Carvalho; Wanderley de Souza
20. EVALUATION OF REAL TIME PCR FOR DIAGNOSIS OF SCHISTOSOMA MANSONI INFECTION
Leonardo F. da Silva; Regina Helena S. Peralta; Mariana Coimbra; Magali G.M. Barreto;
Heloisa W. Macedo; Marta Guimarães Cavalcanti; José Mauro Peralta
21. EVALUATION OF SCHISTOSOMICIDAL POTENTIAL OF EPICLUSIANONE
Aline Pereira Castro; Giulliano Vilela Barros; Marcelo Henrique dos Santos; Liliane
Gançalves Vila Nova; Amanda Esteves Rocha Ferreira; Marcos José Marques; Raquel
Lopes Martins Souza
22. EVALUATION OF THE GENOMIC DNA OF SCHISTOSOMA MANSONI IN BLOOD SAMPLES OF
MICE, BEFORE AND AFTER TREATMENT WITH PRAZIQUANTEL
Wilma Patrícia de Oliveira Santos Bernardes; Fábio Ribeiro; Nilton Barnabé Rodrigues;
Paulo Marcos Zech Coelho; Naftale Katz; Neusa Araújo
23. EVALUATION OF THE PREVALENCE OF SCHISTOSOMIASIS IN THE MUNICIPALITIES OF THE
HEALTH MANAGEMENT OFFICE OF ALFENAS- MG, BRAZIL
Dener Pádua Pimenta; Danilo Ribeiro Ferraz; Rosângela Siqueira Vieira; Liliane Gonçalves
Vila Nova; Letícia Gonçalves Resende Ferreira; Rubens dos Santos Vieira Júnior; Raquel
Lopes Martins Souza
31
24. ACTIONS PERFORMED BY SECRETARIA ESTADUAL DE SAUDE FOR THE SURVEILLANCE OF
SCHISTOSOMIASIS IN THE OF STATE RIO DE JANEIRO, BRAZIL.
Eduardo Faraj Delmas; Sergio Pereira Cunha; Cristina Maria Giordano Dias
25. APPROACH ON THE TRANSMISSION, THE SYMPTOMS AND PROPHYLAXIS OF PARASITES
DISEASES MOST COMMON IN MARANHÃO STATE
Marjane Soares Ferreira; Selma Patrícia Diniz Cantanhede; Andiara Garcez de Souza
Silva; Hallyne Davinck Mesquita Moreira; Gisele Cavalcante Morais; Luciana Patrícia
Lima Alves; Nêuton Silva-Souza
26. HEALTH PROMOTING BY SCHOOLS AND COMMUNITY - SCHISTOSOMIASIS. PERNAMBUCO
2010
Eduardo Albuquerque
27. RURAL SCHOOL-BASED PROGRAM: SCHISTOSOMIASIS PREVENTION AMONG CHILDREN
IN AN EDEMIC AREA IN, MINAS GERAIS STATE, BRAZIL
Indira Simões Martins; Leonardo Ferreira Matoso; Maria Flávia Gazzinelli; Cristiano Lara
Massara; Helmut Kloos; Andrea Gazzinelli; Dener Carlos Dos Reis
28. SOCIAL REPRESENTATION OF EVERYDAY EVENTS ON THE MARGINS OF THE PARDO
RIVER. IMPLICATIONS FOR THE HEALTH-DISEASE PROCESS, WITH EMPHASIS ON
SCHISTOSOMIASIS
Glêissia Amorim Tigre; Lúcia Alves de Oliveira Fraga
29. COMPARATIVE STUDY OF THE TUBERCLES OF MALE SCHISTOSOMA MANSONI USING
CONFOCAL LASER AND SCANNING ELECTRON MICROSCOPY
Roberto Carlos Ferreira João; Cléria C. Mello-Silva; Viviane da Silva Costa; Cláudia Portes
Santos
30. EARLY PRODUCTION OF REACTIVE OXYGEN SPECIES (ROS) BY BIOMPHALARIA
TENAGOPHILA HEMOCYTES FROM THE TAIM STRAIN, WHICH IS RESISTANT TO THE
INFECTION WITH SCHISTOSOMA MANSONI
Cintia Ap. de Jesus Pereira; Emília Souza Araújo; Leonardo Rodrigues; Luciano S. A.
Capettini; Michelle C. de Rezende; Florence M. Rosa; Vanessa Fernandes Rodrigues; Paulo
Marcos Zech Coelho; Ary Corrêa Junior; Deborah Negrão-Corrêa
31. EVALUATION MOLLUSCICIDE CHAMAESYCE HIRTA MILL SP. FOR BIOMPHALARIA
GLABRATA
Luciana Patricia Lima Alves; Marta Martins Almeida; Selma Patricia Diniz Cantanhede;
Joedilsa Sena; Hallyne Davinck Mesquita Moreira; Andiara Garcez de Souza Silva; Marjane
Soares Ferreira; Nêuton Silva-Souza; Jackson Ronie Sá da Silva; Alessandra Leda Valverde
32. EVALUATION OF NATURAL SCHISTOSOMA MANSONI INFECTION OF BIOMPHALARIA
STRAMINEA IN THE PARQUE ESTADUAL DO SUMIDOURO, LAGOA SANTA, MINAS GERAIS,
BRAZIL
Amanda Cardoso De Oliveira Silveira; Érica Oliveira Dias; Mônica Johanna Lukestik;
Sabrina Lúcia Marçal Dos Santos; Luiz Alberto Dolabela Falcão; Liana Konovaloff JannottiPassos
32
33. EVALUATION OF THE SPECIES AND SPACE DISTRIBUTION OF BIOMPHALARIA SP. IN THE
MUNICIPALITY OF ILHA DAS FLORES, SERGIPE, BRAZIL
Delmany Moitinho Barboza; Verônica de Loudes Sierpe Jeraldo; Claudia Moura de Melo;
Mario Adriano dos Santos; Nataly Cardoso Santos; Cangjie Zhang; Guilherme Loureiro
Werneck; José Antônio Pacheco de Almeida; Amélia Ribeiro de Jesus
34. EXPERIMENTAL INFECTION OF SUBSPECIES BIOMPHALARIA TENAGOPHILA GUAIBENSIS
(MOLLUSCA: PLANORBIDAE) WITH SCHISTOSOMA MANSONI SAMBOM, 1907
Lidiane Bento Braga; Liana Konovaloff Jannotti-Passos; Omar dos Santos Carvalho;
35. GIS CHARACTERIZATION OF THE ECOLOGICAL NICHES OF SCHISTOSOMIASIS VECTORS
FROM THE SUB-BASIN DOIS RIOS RIVER, RIO DE JANEIRO STATE, BRAZIL
Pablo Menezes Coelho; Paula Thaise Bermudez dos Reis; Monica Ammon Fernandez;
Silvana Carvalho Thiengo
36. INFECTION OF BIOMPHALARIA GLABRATA BY SCHISTOSOMA MANSONI IN THE
SURROUNDING SUBURBS OF SÃO LUÍS, MARANHÃO
Andiara Garcez de Souza Silva; Luciana Patrícia Lima Alves; Selma Patrícia Diniz
Cantanhede; Marjane Soares Ferreira; Nêuton Silva-Souza
37. MALACOLOGICAL SPECIES OF BIOMPHALARIA OF CITIES IN THE JURISDICTION IN THE
MANAGEMENT OF REGIONAL HEALTH ALFENAS-MG, BRAZIL
Rubens dos Santos Vieira Júnior; Rosangela Vieira Siqueira; Dener Pádua Pimenta; Liliane
Gonçalves Vila Nova; Letícia Gonçalves Resende Ferreira; Danilo Ribeiro Ferraz; Raquel
Lopes Martins Souza
38. MALACOLOGY AND ENVIRONMENTAL ASSESSMENT OF URBAN OUTBREAKS IN THE
METROPOLITAN AREA OF ARACAJU, SERGIPE, BRAZIL
Daniel Santos Oliveira; Veronica de Lourdes Sierpe Jeraldo; Cláudia Moura de Melo;
Álvaro Silva Lima; Sirleide Pereira Farias; Camila Dantas de Carvalho; Vanessa Bispo
Santos; Alexandro Carvalho Silva
39. POPULATION DYNAMICS OF BIOMPHALARIA AMAZONICA POTENTIAL VECTOR OF
SCHISTOSOMA MANSONI AND THE EXOTIC SNAIL MELANOIDES TUBERCULATUS IN APM
RESERVOIR, MATO GROSSO, BRAZIL
Monica Ammon Fernandez; Aline Carvalho de Mattos; Silvana Carvalho Thiengo
40. POSITIVE FOR SCHISTOSOMA MANSONI ON BIOMPHALARIA GLABRATA COLLECTED IN
NEIGHBORHOOD COROADINHO, SÃO LUÍS – MA
Luciana Patrícia Lima Alves; Andiara Garcez de Souza Silva; Selma Patrícia Diniz
Cantanhede; Marjane Soares Ferreira; Marta Martins Almeida; Hallyne Davinck Mesquita
Moreira; Nêuton Silva-Souza; Alessandra Leda Valverde; Jackson Ronie Sá da Silva
41. PROCESS MANEGEMENT AIMING THE IMPROVEMENT OF B. GLABRATA COLONY
MAINTANCE AT MOLUSCÁRIO DO LABORATÓRIO DE ESQUISTOSSOMOSE EXPERIMENTAL/
IOC/FIOCRUZ TO ENSURE THE EFFECTIVENESS OF SCIENTIFIC RESEARCH IN DEVELOPING
Ronaldo de Carvalho Augusto; Patrícia Machado Pinto; Clélia Christina Correa Mello Silva
33
42. SUSCEPTIBILITY OF BIOMPHALARIA TENAGOPHILA FROM ILHA GRANDE, ANGRA DOS
REIS, RJ, TO INFECTION OF SCHISTOSOMA MANSONI
Monica Ammon Fernandez; Marta Chagas Pinto; Silvana Carvalho Thiengo
43. THE MOLLUSCICIDAL ACTION OF THE LATEX OF EUPHORBIA SPLENDENS VAR. HISLOPII
AND THE SCHISTOSOMA MANSONI INFECTION ON THE CONCENTRATIONS OF TOTAL
PROTEINS AND NITROGEN PRODUCTS IN BIOMPHALARIA GLABRATA INFECTED WITH
SCHISTOSOMA MANSONI DURING LATEX SOLUTION
Mariana Gomes Lima; Ronaldo de Carvalho Augusto; Maurício Carvalho de Vasconcellos;
Clélia Christina C.Mello Silva; Jairo Pinheiro
44. AN OUTBREAK OF ACUTE SCHISTOSOMIASIS IN THE SOUTHEAST OF BRAZIL: I.
EPIDEMIOLOGICAL ASPECTS
Sandra Costa Drummond; Izabela Voieta; Alba Otoni; Silvana Romano; José F.Vilela;
Jeann Vilela; Fernando J. Silva; Wenderson Bassoli; Patrícia Passos Botelho; Carla Maria
Ligeiro; Carlos Maurício Figueiredo Antunes; José Roberto Lambertucci
45. ASPECTS RELATED TO THE DEATH BY SCHISTOSOMIASIS, OBTAINED FROM INVESTIGATIONS
WITH FAMILY IN JABOATAO DOS GUARARAPES-PE, JUNE 2010
Francine Nesello; Tania Gomes de Carvalho; José Holanda dos Santos Neto; Rodriga
Maria Zovkar de Souza; José Alexandre Menezes da Silva
46. CARBOHYDRATE METABOLISM IN BIOMPHALARIA GLABRATA INFECTED WITH
SCHISTOSOMA MANSONI AND EXPOSED TO LATEX FROM EUPHORBIA SPLENDENS VAR.
HISLOPII DURING ITS DEGRADATION
Mariana Gomes Lima; Ronaldo de Carvalho Augusto; Clélia Christina Correa Mello Silva;
Jairo Pinheiro
47. CO-INFECTION OF SCHISTOSOMA MANSONI WITH HIV/AIDS IN ENDEMIC POPULATION IN
THE STATE OF ALAGOAS
Danielle Correia Gama; Rozangela Maria de Almeida Fernandes Wyszomirska; Maria
Sônia Correia Alves; Luciano Fernandes Pereira; Roberta Maria Pereira Albuquerque de
Melo; Dńarte Hermogenes Bastos; Hugo Cabral Tenório; Andrei Leite Gazzaneo
48. EFFECT OF PHYSALIS ANGULATA (CAMAPÚ) EXTRACT AND FRACTION IN EXPERIMENTAL
SCHISTOSOMIASIS MANSONI
José Augusto Albuquerque dos Santos; Ivone Maria Ribeiro; Therezinha Coelho Barbosa
Tomassini; Neusa Araújo; Ana Karine Sarvel de Castro; Luis Cláudio Muniz-Pereira;
Antonio Henrique Almeida de Moraes Neto; Naftale Katz
49. EFFECT OF TREATMENT WITH PRAZIQUANTEL IN COGNITIVE PERFORMANCE OF
CHILDREN INFECTED BY S. MANSONI IN A RURAL AREA OF VALE DO JEQUITINHONHA,
MINAS GERAIS
Márcia Christina Caetano de Souza; Mery Natali Silva Abreu; Humberto F.O. Quites;
Leonardo F. Matoso; Jorge Gustavo Velasquez-Melendez; Helmut Kloos; Rodrigo CorreaOliveira; Andrea Gazzinelli
34
50. EPIDEMIOLOGICAL ASPECTS AND GEOGRAPHICAL DISTRIBUTION OF SCHISTOSOMIASIS
AND HELMINTHES IN SERGIPE STATE ACCORDING TO THE DATA OF THE SCHISTOSOMIASIS
CONTROL PROGRAM (PCE)
Carla Virginia Vieira Rollemberg; Cybele Maria Bomfim Santos; Marília B. Lemos Silva;
Acácia M.B. Souza; Ângela Maria da Silva; José Antônio Pacheco de Almeida; Roque
Pacheco de Almeida; Amélia Ribeiro de Jesus
51. EPIDEMIOLOGICAL ASPECTS OF SCHISTOSOMIASIS IN THE LANDLESS RURAL
Genilde Gomes de Oliveira; Angela Maria da Silva; Izabel Cristina Okuveira Lima; Jenisson
Oliveira Conceição
52. EPIDEMIOLOGICAL INVESTIGATION ON AN AUTOCHTHONOUS CASE OF SCHISTOSOMIASIS
IN THE APIPUCOS LAGOON, CITY OF RECIFE, PERNAMBUCO
Constança Simões Barbosa; Elainne Christine Souza Gomes; Fabio Melo; Onicio Batista
Leal Neto; Louisiana Quinino; Mariana Melo; Lidya Angelo Bezerra; Karina Cocenição
Araujo
53. EPIDEMIOLOGICAL STUDY OF SCHISTOSOMIASIS MANSONI INFECTION IN TWO COUNTIES
OF JEQUITINHONHA AND RIO DOCE VALLEYS IN MINAS GERAIS
Maria José Conceição; Aline Eduardo Carlôto; Fabiana Euzébio Gonçalves Euzébio; Eric
Vinaud de Melo; Iran Mendonça da Silva; Natália Machado da Silva; José Borges Pereira;
Nelson Gonçalves Pereira; José Rodrigues Coura
54. ESTIMATIVE OF SCHISTOSOMIASIS POSITIVE INDEX FOR MINAS GERAIS STATE, BRAZIL,
USING MULTIPLE REGRESSION
Ricardo José de Paula Souza e Guimarães; Corina da Costa Freitas; Luciano Viera Dutra;
Sandra da Costa Drummond; Guilherme Corrêa de Oliveira; Omar dos Santos Carvalho
55. EVALUATE THE EFFECT OF AGE IN REDUCING THE PREVALENCE OF SCHISTOSOMIASIS,
USING COHORT ANALYSIS
Erica Magueta; Michele Mendonça; Alessandra Miranda; Rosiane Araújo; Alda Maria
Soares Silveira; Elaine Speziali; Elizabeth Castro Moreno; Lucia Alves de Oliveira Fraga
56. EVALUATION OF THE SCHISTOSOMIASIS SURVEILLANCE SYSTEM IN ALAGOAS STATE,
BRAZIL, FROM 2004 TO 2008
Gilmara Lima Nascimento; Maria José Menezes2; Jean Lúcia dos Santos; Wildo
Navegantes de Araujo
57. FACTORS ASSOCIATED WITH MAINTENANCE OF SCHISTOSOMIASIS IN THE FOREST ZONE
OF PERNAMBUCO
Verônica Santos Barbosa; Karina Conceição Araújo; Tadeu Rodrigues; Constança Simões
Barbosa
58. GEOSPATIAL ANALYSIS ON HUMAN CASES OF MANSON’S SCHISTOSOMIASIS IN A
HORTICULTURAL COMMUNITY IN THE ZONA DA MATA, PERNAMBUCO, BRAZIL
Onicio Batista Leal Neto; Elainne Christine de Souza Gomes; Fabrício Andrade Martins
Esteves; Thiago Yury Cavalcanti Galvão; Karina Conceição o Araújo; Constança Simões
Barbosa
35
59. GEOSPATIAL DISTRIBUTION OF VECTOR MOLLUSCS FOR SCHISTOSOMIASIS ON THE
COAST OF PERNAMBUCO AND THE METROPOLITAN REGION OF RECIFE
Constança Simões Barbosa; Onicio Batista Leal Neto; Manuel Amarista Sevilha; Fabio
Melo; Jones Albuquerque; Elainne Christine Souza Gomes; Reinaldo Souza dos Santos;
60. HIV INFECTION AND ENDEMIC DISEASES: CASES‘S DISTRIBUTION OF AIDS AND
SCHISTOSOMIASIS IN ALAGOAS
Arthur Maia Paiva; Jean Lúcia dos Santos
61. HOSPITALIZATION AND MORTALITY OF SCHISTOSOMIASIS MANSONI IN CEARA STATE,
BRAZIL, FROM 2000 TO 2009
Vivian da Silva Gomes; Ricristhi Gonçalves de Aguiar Gomes; Alberto Novaes Ramos
Junior; Carlos Henrique Morais de Alencar; Jorg Heukelbach; Fernando Schemelzer
Moraes Bezerra
62. IDENTIFICATION OF MAIN ENTEROPARASITOSIS AND SCHISTOSOMIASIS MANSONI
IN STUDENTS OF TWO SCHOOLS PERIPHERAL OF SANTANA OF IPANEMA, STATE OF
ALAGOAS, BRAZIL
Márcio Bezerra Santos; Erlon Oliveira dos Santos; Karina Conceição Gomes Machado de
Araújo
63. LONGITUDINAL STUDY ON SCHISTOSOMIASIS AND SOCIOECONOMIC VARIABLES IN A
RURAL AREA IN MINAS GERAIS STATE, BRAZIL
Humberto Ferreira de Oliveira Quites; Helmut Kloos; Leonardo Ferreira Matoso; Mery
Natali Silva Abreu; João Paulo Amaral Haddad; Rodrigo Corrêa-Oliveira; Andréa Gazzinelli
64. MOLLUSCICIDAL ACTIVITY OF HEXANE AND DICHLOROMETHANE FRACTIONS OF
ETHANOL EXTRACT OF GINGER (ZINGIBER OFFICINALE) ON THE BIOMPHALARIA
GLABRATA, INTERMEDIATE HOST OF SCHISTOSOMA MANSONI
Teiliane Rodrigues Carneiro; Antonio Gomes da Silva Neto; James Almada da Silva;
Ana Lúcia de Paula Hanemann; Paulo Cesar Vieira; Renata de Sousa Alves; Fernando
Schemelzer de Moraes Bezerra
65. 14056 - MOLLUSCICIDE ACTIVITY OF THE ETHANOL EXTRACT OF THE LEAF OF ANACARDIUM
OCCIDENTALE L. ABOUT BIOMPHALARIA GLABRATA (SAY, 1818), INTERMEDIATE HOST OF
SCHISTOSOMA MANSONI (SAMBON, 1907)
Teiliane Rodrigues Carneiro; Morgana M. O. Barboza; Márcia M. M. Marques; Ana R. A.
Silva; Sérvio Q. Júnior; Fernando Schemelzer de Moraes Bezerra; Maria I. F. Guedes
66. MORPHOLOGICAL CHANGES IN SCHISTOSOMA MANSONI ADULT WORMS RECOVERED
FROM NITRIC OXIDE SYNTHASE DEFICIENT MICE FED A LOW-PROTEIN DIET
Renata Heisler Neves; Renata Pinto Ramos; Silvia M. L. Montenegro; Eridan Medeiros
Coutinho; José Roberto Machado-Silva
67. MORPHOLOGY OF “BREVIFURCATE APHARYNGEATE CERCARIA” (SCHISTOSOMATIDAE)
AND THE RECORDS OF ITS OCCURRENCE IN BRAZIL
Aline Carvalho de Mattos; Bruno Guimarães Lopes; Monica Ammon Fernandez; Fábio
Fiebrig Buchmann; Silvana Carvalho Thiengo
36
68. MUNICÍPIOS DO PARANÁ ONDE FORAM ENCONTRADAS AS TRÊS ESPÉCIES HOSPEDEIRAS
INTERMEDIÁRIAS DO SCHISTOSOMA MANSONI NO MESMO MUNICÍPIO, NUM UNIVERSO
DE 210 PESQUISADOS
Jaqueline Finau; Antonio P. Carvalho; Dourival Ribeiro da Silva
69. NATURAL INFECTION OF SCHISTOSOMIASIS MANSONI INTERMEDIATE HOSTS IN THE
COAST OF PERNAMBUCO
Mariana Izabel Sena Barreto de Melo; Constança Simões Barbosa; Fábio Lopes de Melo;
Manuel Amarista Sevilla
70. O PROGRAMA DE CONTROLE DA ESQUISTOSSOMOSE EM DOIS MUNICÍPIOS DA ZONA DA
MATA DE PERNAMBUCO: ANALISANDO A IMPLANTAÇÃO DAS AÇÕES
Louisiana Regadas de Macedo Quinino; Isabella Chagas Samico; Constança Simões
Barbosa
71. POLYPARASITISM BETWEEN SCHISTOSOMA MANSONI AND INTESTINAL PARASITES:
DETERMINANTS AND GEOSTATISTIC
Carla Virginia Vieira Rollemberg; Fábio Jorge Ramalho Amorim; Karla Carolline Vieira
Rollemberg; Marília M. B. L. Silva; Acácia M. B. Souza; Enaldo Vieira; Evan W. Secor;
Guilherme Loureiro Werneck; José Antônio Pacheco de Almeida; Amélia Ribeiro de Jesus
72. SCHISTOSOMA MANSONI PREVALENCE IN STUDENTS AS AN INDICATOR OF INFECTION IN
THE GENERAL POPULATION IN A HYPERENDEMIC RURAL AREA IN MINAS GERAIS STATE
Kellen Rosa Coelho; Martin Johannes Enk; Leonardo Ferreira Matoso; Mery Natali Silva
Abreu; Rodrigo Corrêa Oliveira; Helmut Kloos; Andréa Gazzinelli
73. SCHISTOSOMIASIS AND ECOSYSTEM APPROACH TO HEALTH: A SYSTEMATIC REVIEW
Marisa da Silveira Soares; Margareth Maria Lessa Gonçalves; Magali Gonçalves Muniz
Barreto;
74. SCHISTOSOMIASIS IN PATIENTS ATTENDED AT THE HOSPITAL OF THE FEDERAL UNIVERSITY
OF ALAGOAS IN MACEIO (ALAGOAS- BRAZIL)
Thiago André Alves Fidelis; Thiago de Oliveira Assis; Janira Lúcia Assumpção Couto;
Rozangela Maria de Almeida Fernandes Wyszomirska
75. SCHISTOSOMIASIS IN THE TOURIST CITY OF HOLAMBRA (SÃO PAULO, BRAZIL), FROM
1993 TO 2008
Marisa da Silveira Soares; Celia Maria Thomé; Cesar Luiz Pinto Ayres Coelho da Silva;
Claudia Portes Santos Silva; Magali Gonçalves Muniz Barreto; Denise de Assuncão
Borges; Rita Maria Silva; Cybele Gargioni; Vera L. M. Oliveira; Herminia Y. Kanamura
76. SCHISTOSOMIASIS MANSONI IN CHILDREN AND YOUTH OF A MUNICIPAL DISTRICT OF
ALAGOAS (BRAZIL)
Janira Lúcia Assumpção Couto; Michelle Martins da Silva; Nathalie Louise Santos de
Oliveira; Elaine Cristina e Silva; Mykaella Andrade de Araujo; Jefferson Alain da Costa
Ferreira; Thiago Andre Alves Fidelis; Alinne Schristina da Silva; Pedro Filippe Simões
Santos; Francisnaldo Emanuel Nunes P. Torres
37
77. SNAIL SPACIAL DISTRIBUTION IN UBERLANDIA - MG‘S URBAN ZONE
Amaral Alves de Souza; Alessandro Ambrosio dos Reis; Lara Lane de Oliveira; Adalberto
Albuquerque Pajuaba Neto; Elisângela de Azevedo Silva Rodrigues; Márcia Beatriz
Cardoso de Paula
78. SPATIAL ANALYSIS OF SCHISTOSOMIASIS HUMAN CASES AT SANTA MARIA
NEIGHBORHOOD, ARACAJU-SERGIPE
Karina Conceição Gomes Machado de Araújo; Helder Barreto Silva Junior; Roseli La Corte
dos Santos; Andréa Valença Cardoso
79. SPATIAL ANALYSIS OF SCHISTOSOMIASIS VECTOR FOCI IN SANTA MARIA NEIGHBORHOOD,
ARACAJU-SE
Tiago Pinheiro Vaz de Carvalho; Roseli La Corte dos Santos; Constança Simões Barbosa;
Karina Conceição Gomes Machado de Araújo
80. SPATIAL DISTRIBUTION OF DEATHS FROM SCHISTOSOMIASIS IN PERNAMBUCO, FROM
2005 TO 2008
Bárbara Morgana da Silva; Silvânia AA Lima; Eduardo N Albuquerque; Neusa E
Magalhães; Nara Pedrosa Arruda
81. SPATIAL DISTRIBUTION OF SCHISTOSOMA MANSONI INFECTION IN ADOLESCENTS
UNDERGOING CHEMOTHERAPY WITH PRAZIQUANTEL IN AN ENDEMIC AREA OF
SCHISTOSOMIASIS IN PERNAMBUCO, BRAZIL
Aline Favre Galvão; Oswaldo Gonçalves Cruz; Tereza Cristina Favre; Otávio Sarmento Pieri
82. SPATIAL-TEMPORAL ANALYSIS ON FOCI OF SCHISTOSOMIASIS TRANSMISSION IN PORTO
DE GALINHAS, PERNAMBUCO: A PROCESS OF ENDEMIZATION ?
Elainne Christine de Souza Gomes; Onicio Batista Leal Neto; Karlla Priscila Monteiro
Viana; Isabela Regina Alvares da Silva Lira; Rafael Cesar Lima Pedroso de Andrade; Diego
Leandro Reis da Silva Fernandes; Julyana Viégas Campos; Hilda Carla Moura dos Santos;
83. THE CHANGING RISK IN THE EPIDEMIOLOGIC PROFILE OF SCHISTOSOMIASIS IN
RONDÔNIA STATE
Flávia Serrano Batista; Carlos Antônio Amante; Sônia Maria Dias de Lima; Maria de
Nazaré Reis Alves
84. THE COMPLEXITY OF SCHISTOSOMIASIS IN HOLAMBRA (SÃO PAULO, BRAZIL): CHECKING
BASES FOR AN ECOSYSTEM APPROACH TO HEALTH
Marisa da Silveira Soares; Marcela C. Mansano; Magali G. M. Barreto; Marcelo F.S. Porto
85. TREND OF THE SCHISTOSOMIASIS MANSONI IN MARANHÃO STATE: 1997 TO 2003
Selma Patricia Diniz Cantanhede; Inês Echenique Mattos; Aldo Pacheco Ferreira
86. USEFULNESS OF THE NETWORK OF CONTACTS FOR THE DETECTION OF SCHISTOSOMIASIS
IN LOW INTENSITY TRANSMISSION FOCUS, ESTEIO, RIO GRANDE DO SUL
Daniela Martins Toniolli; Maria Ceci Salcedo Botelho; José Antônio Silveira; Carlos Graeff
Teixeira
38
07/10 (18h00-19h00)
LOCaL: fOYER
Imunopatologia, Biologia Molecular e Bioquímica, diagnóstico, tratamento e Clínica
(Immunopathology, Molecular Biology, Biochemistry, Diagnosis, Treatment and Clinical)
01. EVALUATION OF TREATMENT WITH ANTIBODIES ANTI CO-STIMULATORY MOLECULES ON
CELLULAR RESPONSE OF SHISTOSOMIASIS
Laís Cristina de Souza; Naiara Naiana Dejani; Joice Margareth de Almeida Rodolpho;
Sandra Regina Pereira de Oliveira; Débora Meira Neris; Vitor Leão; Ricardo de Oliveira
Correia; Carlos Alberto Soares; Auro Nomizo; Heloísa Sobreiro Selistre de Araújo;
Fernanda de Freitas Anibal
02. EXPERIMENTAL ANALYSIS OF THE IMIDAZOLINE DERIVATIVE 3-BENZYL-5-(4-CHLOROARILAZO)-4-IMIDAZOLIDINE-2-ONE AGAINST ADULT WORMS OF SCHISTOSOMA
MANSONI
Andréa Cristina Aplinário Silva; Juliana Kelle de Andrade Lemoine Neves; André de Lima
Aires; Anekecia Lauro da Silva; Tiago Moreira Alves Feitosa; Renata Alexandre Ramos
Silva; Maria do Carmo Alves de Lima; Ivan da Rocha Pitta; Suely Lins Galdino; Mônica
Camelo Pessoa de Azevedo Albuquerque
03. EXPERIMENTAL CHEMOTHERAPY IN SCHISTOSOMIASIS USING RUTHENIUM NITRIC
OXIDE DONOR
Francine Nesello; Jean Jerley Nogueira da Silva; Carolina Panis; Ivete Conchon Costa;
Maria Claudia Noronha Dutra de Menezes; Francisco José de Abreu Oliviera; Thiago
Mattar Cunha; Maria Angélica Ehara Watanabe; Wander Rogério Pavanelli
04. EXPERIMENTAL STUDY OF SCHISTOSOMA MANSONI ISOLATE FROM PATIENT WITH
THERAPEUTIC FAILURE TO PRAZIQUANTEL
Maria José Conceição; Eric Vinaud de Melo; Aline Eduardo Carlôto; Natália Machado
da Silva; Iran Mendonça da Silva; Isabel Cristina Melo Mendes; Viviane Pereira; Nelson
Gonçalves Pereira; José Borges-Pereira
05. IMPROVING METHODS OF INDIRECT ELISA USING ANTI-IGG ANTIGEN SOLUBLE ADULT
WORM (SWAP), SOLUBLE EGG ANTIGEN (SEA) AND TEGUMENT OF SCHISTOSOMULA
ANTIGEN (SMTEG) IN SERA OF PATIENTS THE CHRONIC PHASE OF SCHISTOSOMIASIS
MANSONI
Watson Herman Martins; Rafaella Fortini; Elizandra Giani Ribeiro; Vanessa Silva Moraes;
Edward Jose de Oliveira; Paulo Marcos Zech Coelho
06. IN VITRO SCHISTOSOMICIDAL ACTIVITY OF (-)-6,6’-DINITROHINOKININ AGAINST
SCHISTOSOMA MANSONI
Lizandra Guidi Magalhães; Ana Carolina Pereira; Thais Coelho Lima; Karen C S Rezende;
Jairo K Bastos; Márcio L A e Silva; Vanderlei Rodrigues; Ademar Alves da Silva Filho;
Wilson Roberto Cunha
39
07. IN VIVO EVALUATION OF THE IMIDAZOLINE DERIVATIVE 3-BENZYL-5-(4-CHLOROARILAZO)-4-IMIDAZOLIDINE-2-ONE AGAINST EVOLUTIVE FORMS OF SCHISTOSOMA
MANSONI
Amanda Caroline Malaquias de Farias; Juliana Kelle de Andrade Lemoine Neves; André
de Lima Aires; Tiago Moreira Alves Feitosa; Renata Alexandre Ramos da Silva; Maria do
Carmo Alves de Lima; Ivan da Rocha Pitta; Suely Lins Galdino; Mônica Camelo Pessoa de
Azevedo Albuquerque
08. IN VIVO EVALUATION OF THE IMIDAZOLINE DERIVATIVE LPSF/RZS-5 AGAINST ADULT
WORMS OF SCHISTOSOMA MANSONI
Juliana Kelle de Andrade Lemoine Neves; André de Lima Aires; Anekécia Lauro da Silva;
Tiago Moreira Alves Feitosa; Renata Alexandre Ramos Silva; Maria do Carmo Alves
de Lima; Ivan da Rocha Pitta; Suely Lins Galdino; Mônica Camelo Pessoa de Azevedo
Albuquerque
09. IN VIVO SCHISTOSOMICIDAL EFFECT OF THE IMIDAZOLINE DERIVATIVE 5-(4-FLUORARILAZO)-3-BENZYL-4-TIOXO-IMIDAZOLIDINE-2-ONE AGAINST ADULT WORMS OF
SCHISTOSOMA MANSONI
Juliana Kelle de Andrade Lemoine Neves; André de Lima Aires; Tiago Moreira Alves
Feitosa; Anekecia Lauro da Silva1; Renata Alexandre Ramos Silva; Maria do Carmo Alves
de Lima1; Ivan da Rocha Pitta1; Suely Lins Galdino1; Mônica Camelo Pessoa de Azevedo
Albuquerque
10. INHIBITORY EFFECTS OF HARPAGOPHYTUM PROCUMBENS ON DISPOSAL OF EGGS
DURING INFECTION WITH SCHISTOSOMA MANSONI
Joice Margareth de Almeida Rodolpho; Sandra Regina Pereira de Oliveira; Laís Cristina
de Souza; Naiara Naiana Dejani; Débora Meira Neris; Ricardo de Oliveira Correia; Jame‘s
Almada da Silva; Paulo Cesar Vieira; Fernanda de Freitas Anibal
11. OCCURRENCE OF SCHISTOSOMIASIS IN THE MUNICIPALITIES OF SÃO BENTO AND SÃO
LUÍS, MARANHÃO STATE, IN THE PERIOD 1998 TO 2008
Selma Patrícia Diniz Cantanhede; Andiara Garcez de Souza Silva; Luciana Patrícia Lima
Alves; Marjane Soares Ferreira; Nêuton Silva-Souza
12. PCR ASSAY OF HUMAN URINE SAMPLES FOR THE DIAGNOSIS OF SCHISTOSOMIASIS
MANSONI
Nilton Barnabé Rodrigues; Guilherme Oliveira e Silva; Martin J. Enk
13. PORTAL HYPERTENSION IN SCHISTOSOMIASIS MANSONI: COMPLICATIONS OF ELECTIVE
SURGICAL TREATMENT
Izabela Voieta; Andy Petroianu; Vivian Resende; Kelly Rodrigues; Vitor Freitas Fontes;
Juliana Papatella; Carlos Mauricio de Figueiredo Antunes; José Roberto Lambertucci
14. PRELIMINARY STUDY OF PULMONARY FUNTIONS TESTS IN PATIENTS WITH MANSONI’ S
SCHISTOSOMIASIS
Rita de Cassia dos Santos Ferreira; Amanda Medeiros Gomes da Silva; Ana Lúcia Coutinho
Domingues; Angela Pontes Bandeira
40
15. PROFILE OF SPECIFIC HUMORAL RESPONSES AGAINST SCHISTOSOMIASIS IN AN ENDEMIC
RURAL AREA OF MINAS GERAIS STATE, BRAZIL: A LONGITUDINAL STUDY
Leonardo Ferreira Matoso; Ricardo Toshio Fujiwara; Humberto Ferreira de Oliveira
Quites; Lorena Junia de Souza Santos; Luciana Lisboa Mota e Castro; Mery Natali Silva
Abreu; Helmut Kloos; Rodrigo Correa-Oliveira; Andréa Gazzinelli
16. SCHISTOSOMAL MYELORADICULOPATHY: A 10-YEAR FOLLOW-UP STUDY
Sílvio Roberto Sousa-Pereira; Thiago Cardoso Vale; Paloma Fonseca; Tânia Antunes
Carvalho; Luciana Cristina dos Santos Silva; Izabela Voieta; Carlos Maurício de Figueiredo
Antunes; José Roberto Lambertucci
17. SCHISTOSOMICIDAL ACTIVITY AND TEGUMENTAL ALTERATIONS INDUCED BY PIPLARTINE
ON SCHISTOSOMULA AND ADULT FLUKES OF SCHISTOSOMA MANSONI
Josué de Moraes; Carlos Nascimento; Eliana Nakano; Lydia F. Yamaguchi; Massuo J. Kato;
Toshie Kawano;
18. SCHISTOSOMICIDE EFFECTS OF NEW THIAZOLIDINE DERIVATIVES (EXPERIMENTS IN
VITRO)
Anekécia Lauro da Silva; Sheilla Andrade de Oliveira; Andréia Ferreira de Barros; Veruska
Cíntia Alexandrino de Souza; Antônio Sérgio Alves de Almeida Júnior; Roni Evêncio de
Araujo; Maria do Carmo Alves de Lima; Valéria Rêgo Alves Pereira; Ivan da Rocha Pitta;
Suely Lins Galdino
19. THE IMPORTANCE OF EARLY DIAGNOSIS IN NEUROSCHISTOSOMIASIS: CASE REPORTS
Vinícius Martins Alecrim; Tatyana Layla Aguiar Hazin; Joelton Barbosa da Silva; Marcos
Daniel Nigro da Silva; Rodrigo Argenta Toniolo; Ana Virgínia Matos Sá Barreto
20. THIAZOLIDINE DERIVATIVES IN THE TREATMENT OF MICE ACUTELY INFECTED WITH
SCHISTOSOMA MANSONI
Anekécia Lauro da Silva; Sheilla Andrade de Oliveira; Andréia Ferreira de Barros; Veruska
Cíntia Alexandrino de Souza; Jamerson Ferreira de Oliveira; Roni Evêncio de Araujo; Maria
do Carmo Alves de Lima; Ivan da Rocha Pitta; Suely Lins Galdino
21. TREATMENT WITH HARPAGOPHYTUM PROCUMBENS MODULATES EOSINOPHILIA IN
EXPERIMENTAL MODEL OF SCHISTOSOMIASIS
Sandra Regina Pereira de Oliveira; Joice Margareth de Almeida Rodolpho; Ricardo de
Oliveira Correia; Debora Meira Neris; Laís Cristina de Souza; Naiara Naiana Dejane; Paulo
César Vieira; Fernanda de Freitas Anibal
22. ULTRASONOGRAPHICAL ASPECTS OF PERIPORTAL FIBROSIS IN ADOLESCENTS INFECTED BY
SCHISTOSOMA MANSONI IN AN ENDEMIC AREA OF SCHISTOSOMIASIS IN PERNAMBUCO,
BRAZIL
Luciana Carvalho Zani; Otávio Sarmento Pieri; Tereza Cristina Favre; Aline Favre Galvão;
Ana Paula Brás Pereira; Ana Lucia Coutinho Domingues
23. URINARY IMMUNOLOGICAL PROFILE OF HEPATOSPLENIC SCHISTOSOMIASIS PATIENTS
José Roberto Lambertucci; Alba Otoni; Izabela Voieta; Carlos Maurício Antunes; Antônio
Lúcio Teixeira
41
24. VARICEAL UPPER GASTROINTESTINAL BLEEDING IN EMERGENCY HOSPITAL IN RECIFE,
PERNAMBUCO
Thais Cavalcanti de Almeida; Ana Lúcia Coutinho Domingues; José Roberto de Almeida;
Admar Borges da Costa Junior; Adriana G. de Moura; Roberta Cavalcanti de Almeida
25. ACUTE HEPATOSPLENOMEGALY ALTERATIONS IN MICE INFECTED BY SCHISTOSOMA
MANSONI AND TREATED WITH N-ACETYLCYSTEINE AND/OR PRAZIQUANTEL
André de Lima Aires; Tiago Moreira Alves Feitosa; Renata Alexandre Ramos da Silva;
Juliana Kelle de Andrade Lemoine Neve; Zilma Perreira dos Anjos; Giuliana Viegas Schirato;
Elizabeth Malagueno de Santana; Mônica Camelo Pessoa de Azevedo Albuquerque
26. ACUTE NEUROSCHISTOSOMIASIS: A REPORT ON THREE CASES AND REVIEW OF THE
LITERATURE
Thiago Cardoso Vale; Sílvio Roberto de Sousa-Pereira; Izabela Voieta; José Roberto
Lambertucci
27. EFFECTS OF SILYMARIN IN DMSO ON EXPERIMENTAL SCHISTOSOMIASIS MURINE
Fabiana Gonçalves Lino; Hílton Antônio Mata dos Santos; Carolina Carneiro Rocha;
Fabíola Ramos Xavier; Letícia Campos da Costa; Alexandre dos Santos Pyrrho
28. EVALUATION OF THE INTESTINAL MICROBIOTA IN MICE CHRONICALLY INFECTED BY
SCHISTOSOMA MANSONI
André de Lima Aires; Kedma de Magalhães Lima1; Thays Miranda Almeida; Tiago Moreira
Alves Feitosa; Renata Alexandre Ramos Silva; Maria Helena Madruga Lima Ribeiro; Mônica
Camelo Pessoa de Azevedo Albuquerque; Célia Maria Machado Barbosa de Castro
29. HEPATOSPLENIC SCHISTOSOMIASIS: CLINICAL AND LABORATORY CHARACTERISTICS OF
PATIENTS WITH AND WITHOUT PULMONARY HYPERTENSION
Claudio Lemos Morais; Juliana Papatella; Pedro Henrique Lima Prata; Izabela Voieta;
Carlos Maurício Figueiredo Antunes; José Roberto Lambertucci
30. HISTOPATHOLOGICAL AND QUANTITATIVE FEATURES OF SCHISTOSOMAL HEPATIC
GRANULOMAS IN A MOUSE-MODEL OF METABOLIC PROGRAMMING
Christiane Leal Corrêa; Patrícia Cristina Lisboa; José Roberto Machado e Silva; Elaine de
Oliveira; Egberto Gaspar de Moura; Renata Heisler Neves; Regina Maria Figueiredo de
Oliveira; Adriana Cardoso Gomes
31. LIVER HISTOPATHOLOGY OF CHRONIC SCHISTOSOMIASIS IN MICE FED HIGH-FAT DIET
Alba Cristina Miranda de Barros Alencar; Renata Heisler Neves; Marcele Nogueira de
Sousa Trotte; Albanita Viana de Oliveira; Delir Corrêa Gomes; José Roberto Machado-Silva;
32. PRODUCTION OF MABS (MONOCLONAL ANTIBODIES) ANTI-CO-STIMULATORY
MOLECULES FOR THE TREATMENT OF EXPERIMENTAL SCHISTOSOMIASIS MANSONI
Laís Cristina de Souza; Isabela Gobbo Ferreira; Ciro Novaes Lino; Denise Sayuri Calheiros
da Silveira; Bruna Gabriela Silva; André Barros; Naiara Naiana Dejani; Joice Margareth
de Almeida Rodolpho; Sandra Regina Pereira de Oliveira; Débora Meira Neris; Ricardo
Oliveira Correia; Heloísa Sobreiro Selistre de Araújo; Cláudio Alberto Torres Suazo; Auro
Nomizo; Fernanda de Freitas Anibal
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33. PULMONARY HISTOPATHOLOGY OF MICE FED HIGH-FAT DIET ON ACUTE SCHISTOSOMIASIS
MANSONI INFECTION
Vanessa Coelho de Góes; Renata Heisler Neves; Alba Cristina Miranda de Barros Alencar;
Marcele Nogueira de Sousa Trotte; Albanita Viana de Oliveira; Delir Corrêa Gomes; José
Roberto Machado-Silva
34. SCHISTOSOMA ANTIGENS DOWN MODULATE THE INFLAMMATORY IMMUNE RESPONSE
IN VITRO IN PATIENTS INFECTED WITH HTLV-1
Luciane Mota Lima; Silvane Maria Braga Santos; Ricardo Riccio Oliveira; Luciana Santos
Cardoso; Sergio Costa Oliveira; Alfredo Miranda Goes; Alex Loukas; Edgar M. de Carvalho;
Maria Ilma Araújo
35. STUDY OF THE BACTERIAL TRANSLOCATION THROUGH THE INTESTINAL TRACT IN MICE
INFECTED BY SCHISTOSOMA MANSONI
André de Lima Aires; Kedma de Magalhães Lima; Thays Miranda Almeida; Tiago Moreira
Alves Feitosa; Renata Alexandre Ramos Silva; Maria Helena Madruga Lima Ribeiro;
Mônica Camelo Pessoa de Azevedo Albuquerque; Célia Maria Machado Barbosa de Castro;
36. ULTRASONOGRAPHIC AND LABORATORIAL EVALUATION OF CHRONIC SCHISTOSOMIASIS
MANSONI PATIENTS
Ana Virgínia Matos Sá Barreto; Vinícius Martins Alecrim; Tibério B. Medeiros; Edmundo
P. A. Lopes; Ana Lúcia Coutinho Domingues; Sílvia Maria Lucena Montenegro; Clarice
Neuenschwander Lins de Morais
37. UNCOMMON PRESENTATION OF INTESTINAL SCHISTOSOMIASIS: INCREASED MORBIDITY
WITH DISTINCT OUTCOMES AND IMMUNOLOGYCAL PROFILES IN PATIENTS ATTENDING
A TERCIARY HOSPITAL
Marta Guimarães Cavalcanti; Margareth Maria L. Gonçalves; Leonardo F. Silva; José
Mauro Peralta; Kalil Madi
38. ANTIFIBROTIC ACTION OF SILYMARIN ON EXPERIMENTAL SCHISTOSOMIASIS
Hílton Antônio Mata dos Santos; Fabiana Gonçalves Lino; Carolina Carneiro Rocha;
Fabíola Ramos Xavier; Letícia Campos da Costa; Moragana T. L. Castelo-Branco; Claudia
Neto Paiva; Alexandre dos Santos Pyrrho
39. ANTIGEN-SPECIFIC IGG4 IS A MARKER OF RESISTANCE TO SCHISTOSOMIASIS IN ENDEMIC
AREAS IN BAHIA, BRAZIL
Joanemile Pacheco de Figueiredo; Ricardo Riccio Oliveira; Luciana Santos Cardoso; Maria
Cecilia Almeida; Diego Mota Lopes; Leda Maria Alcantara; Edgar Marcelino Carvalho;
Maria Ilma Araujo
40. CYTOKINE PRODUCTION PROFILE ASSOCIATED WITH PERIPORTAL FIBROSIS IN HUMAN
INFECTED WITH S. MANSONI
Gabriela da Silveira e Nunes; Mônica Maria de Almeida; Andréa Teixeira Carvalho; Lúcia
Alves de Oliveira Fraga; Tiago da Costa Morais; Giovanni Gazzinelli; Elaine Speziali; Olindo
Assis Martins-Filho; Rodrigo Correa-Oliveira; Andrea Gazzinelli; Alda Maria Soares Silveira
43
41. ENDOTHELIAL DYSFUNCTION INDUCED BY SCHISTOSOMIASIS
Suellen D‘Arc dos Santos Oliveira; Luciana Silva do Amaral; Luis Eduardo Menezes
Quintas; François Germain Noël; Cláudia Lúcia Martins da Silva
42. EVALUATION OF IMMUNOLOGICAL PARAMETERS ASSOCIATED WITH INFECTION
AND REINFECTION OF DIFFERENT MURINE STRAINS, C57BL-6 AND BALB-C, WITH
SCHISTOSOMA MANSONI
Clarice Carvalho Alves; Tatiane Teixeira de Melo; Patrícia Martins Parreiras; Cristina
Toscano Fonseca
43. HAEMATOLOGICAL AND IMMUNOLOGICAL STUDIES ON THE CO-INFECTION OF
SCHISTOSOMA MANSONI AND PLASMODIUM CHABAUDI USING A RODENT MODEL
Pedro Manuel Ferreira; Henrique Silveira; Ana Júlia Pinto Fonseca Sieuve Afonso; Maria
Amélia Afonso Grácio
44. IDENTIFICATION OF IMMUNODOMINANTS PROTEIN OF SCHISTOSOMA MANSONI
TEGUMENT
Tatiane Teixeira de Melo; Rosiane Aparecida da Silva Pereira; Paulo Marcos Zech Coelho;
Cristina Toscano Fonseca
45. IMMUNOLOGICAL MONITORING OF PATIENTS WITH ACUTE PHASE OF SCHISTOSOMIASIS
MANSONI FOLLOWING SPECIFIC CHEMOTHERAPY WITH PRAZIQUANTEL
Amanda Cardoso de Oliveira Silveira; Matheus Fernandes Costa-Silva; Denise SilveiraLemos; Martin Johannes Enk; Cristiano Lara Massara; Maria Carolina Barbosa Alvarez;
Pedro Henrique Gazzinelli Guimarães; Helena Barbosa Ferraz; Paulo Marcos Zech Coelho;
Rodrigo Corrêa-Oliveira; Olindo Assis Martins-Filho; Giovanni Gazzinelli; Andréa TeixeiraCarvalho
46. IMMUNOMODULATORY EFFECTS OF MENTHA PIPERITA L. TREATMENT ON MURINE
Naiara Naiana Dejani; Laís Cristina de Souza; Vitor Leão; Joice Margareth Rodolpho;
Sandra Regina Pereira de Oliveira; Débora Meira Neris; Ricardo de Oliveira Correia;
Vanderlei Rodrigues; Luis Vitor do Sacramento Silva; Fabiana Rossetto Morais; Lúcia
Helena Faccioli; Heloísa Sobreiro Selistre de Araújo; Fernanda de Freitas Anibal
47. IMUNOMODULATION OF THE IMIDAZOLINE DERIVATIVE 3-BENZYL-5-(4-CHLOROARILAZO)-4-IMIDAZOLIDINE-2-ONE IN THE PREPATENT PHASE OF THE INFECTION BY
SCHISTOSOMA MANSONI
Luiz Henrique de Souza Teixeira; Juliana Kelle de Andrade Lemoine Neves; André de Lima
Aires; Tiago Moreira Alves Feitosa; Renata Alexandre Ramos Silva; Maria do Carmo
Alves de Lima; Ivan da Rocha Pitta; Suely Lins Galdino; Valdenia Maria Oliveira de Souza;
Mônica Camelo Pessoa de Azevedo Albuquerque; Vlaudia Maria Assis Costa
48. INCREASE IN THE LEVELS OF IGG1 REACTIVITY IN CO-INFECTED MICE WITH STRONGYLOIDES
VENEZUELENSIS DURING THE CHRONIC PHASE OF SCHISTOSOMA MANSONI
Michelle Carvalho de Rezende; Núbia Rangel; Deborah Negrão-Corrêa
44
49. PROFILE OF LYMPHOCYTE SUBSETS OF PATIENTS LIVING IN A COMMUNITY OF LOW
ENDEMICITY FOR SCHISTOSOMIASIS MANSONI IN CEARA-BRAZIL
Sara Menezes De Oliveira; Marta Cristhiany Cunha Pinheiro; Teiliane Rodrigues Carneiro;
Bruna Cunha de Alcantara; Elza Maria Morgado Tomaz; Felipe Fernando da Cruz Inacio;
Fernando Schemelzer de Moraes Bezerra; José Ajax Nogueira Queiroz
50. RELATIONSHIP BETWEEN CYTOKINE PRODUCTION AND HEPATIC MORPHOLOGY IN
UNDERNOURISHED MICE INFECTED WITH SCHISTOSOMA MANSONI
Laís Amorim Sacramento; Andréia Ferreira de Barros; Fabiana Letícia da Silva; Vlaudia
Maria Assis Costa; Roni Evêncio de Araújo; Eridan de Medeiros Coutinho; Sheilla Andrade
de Oliveira; Silvia Maria Lucena Montenegro
51. SCHISTOSOMA MANSONI ANTAGONIZES TOXOPLASMA GONDII-INDUCED INTESTINAL
INFLAMMATORY RESPONSE IN CO-INFECTED MICE: ROLE OF TNF-ALPHA AND IL-17.
Luiz Fernando Queiroz; Jacilene Mesquita; Kalil Madi; Heitor S.P. Souza; Marcelo Torres
Bozza; Marta Guimarães Cavalcanti
52. SCHISTOSOMA MANSONI TEGUMENT (SMTEG) MODULATES THE EXPERIMENTAL
ALLERGIC ASTHMA
Cintia Maria Gonçalves da Silva; Gardênia Braz Figueiredo de Carvalho; Fábio A. V.
Marinho; Sérgio Costa Oliveira; Cristina Toscano Fonseca; Lucila Grossi Gonçalves Pacífico
53. THE ROLE OF MACROPHAGE MIGRATION INHIBITORY FACTOR (MIF) IN SEVERE
SCHISTOSOMIASIS
Jailza Lima Rodrigues; Juliana Froeseler Fittipaldi; Adriana Fernandes; Paula Duarte
Eschenazi; Fernanda S. Costa; Emília Souza Araújo; Michelle Carvalho Rezende; José
Roberto Lambertucci; Carlos Maurício Antunes; Mauro Martins Teixeira; Deborah
Negrão-Corrêa
54. ALBUMINURIA IN SCHISTOSOMA MANSONI-INFECTED INDIVIDUALS
Milton Cezar Compagnon; Laís Danielle Ribeiro de Melo; Carmem de Castro Chaves; Ana
Lúcia Coutinho Domingues; Ana Durce Oliveira da Paixão
55. CHARACTERIZATION OF ANTI-INFLAMMATORY ACTIVITY OF A NEW 4,5-DIIDROISOXAZOL
DERIVATIVE AS A POTENTIAL COMPOUND TO DECREASE THE SCHISTOSOMAL
GRANULOMA FORMATION
Amanda Roberta Revoredo Vicentino; Anderson Mendonça Amarante; Vitor Coutinho
Carneiro; Carlos Aleberto Antunes; Cláudia Farias Benjamim; Alcino Palermo de Aguiar;
Marcelo Rosado Fantappié
56. CHARACTERIZATION OF THE GAMMA-SECRETASE COMPLEX IN SCHISTOSOMA MANSONI
AND THE EFFECTS OF ITS INHIBITION ON PARASITE DEVELOPMENT
Lizandra Guidi Magalhães; William de Castro Borges; Renata Guerra Sá; Carla Botelho
Machado; Enyara Rezende Morais; Érika Bueno de Carvalho Moreira; Cláudia Sossai
Soares; Olinda Mara Brigatto; Elenice Aparecida Macedo; Vanderlei Rodriges
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57. CHROMATIN REGULATION IN SCHISTOSOMES AND HISTONE-MODIFYING ENZYMES AS
DRUG TARGETS
Raymond J. Pierce; Stéphanie Caby; Florence Dubois; Julien Lancelot; Jacques Trolet;
Céline Cosseau; Christoph Grunau; Guillaume Mitta; Luiza F.A. Almeida; Leila Nahum;
Guilherme Correa de Oliveira
58. CLONING AND PURIFICATION OF CITOPLASMATIC FORM OF THE ENZYME
SERINEHYDROXIMETHYLTRANSFERASE (SHMT) FROM SCHISTOSOMA MANSONI
Angela Maria Fala; Alexandre Cassago; Ricardo de Marco; Glaucius Oliva; Richard Garrat;
Humberto D´Muniz Pereira;
59. DEVELOPMENTAL REGULATION OF THE SUMO PATHWAY IN SCHISTOSOMA MANSONI
Roberta Verciano Pereira; Fernanada J. Cabral; Liana K. Janotti Passos; Vanderlei
Rodrigues; William Castro Borges; Renata Guerra de Sá
60. SEQUENCE ANALYSIS AND RNA INTERFERENCE STUDIES OF TRANS-SPLICED TRANSCRIPTS
FROM SCHISTOSOMA MANSONI
Marina de Moraes Mourão; Nathalie Dinguirard; Francisco Pereira Lobo; Francisco
Prosdócimi; Timothy P. Yoshino; Glória Regina Franco.
61. IDENTIFICATION OF A CONSERVED DOMAIN RICH IN B CELL EPITOPES WITHIN SOLUBLE
ATP DIPHOSPHOHYDROLASE FROM SCHISTOSOMA MANSONI BY SYNTHETIC PEPTIDES
ANALYSES
Priscila de Faria Pinto; Ana Carolina R. G. Maia; Rita Gabriela P. R. Mendes; Karen T. Flores;
Cristiane Carvalho Campos; Michélia Antônia N. Gusmão; Gabriane Nascimento Porcino;
Michelle Lima Detoni; Marcos Luiz de Oliveira Penido; Maria Aparecida Juliano; Luiz
Juliano; Rodrigo Correa Oliveira; Paulo Marcos Zech Coelho; Eveline Gomes Vasconcelos
62. IMMUNOSTIMULATORY PROPERTIES OF A SYNTHETIC PEPTIDE BELONGING TO THE
SOLUBLE ATP DIPHOSPHOHYDROLASE ISOFORM FROM SCHISTOSOMA MANSONI, AND
IDENTIFICATION OF THIS PROTEIN IN SOLUBLE EGG (SEA) AND WORM (SWAP) ANTIGENS
PREPARATIONS
Michélia Antônia do Nascimento Gusmão; Michelle de Lima Detoni; Rita Gabriela
Pedrosa Ribeiro Mendes; Karen Takaki Flores; Ana Carolina Ribeiro Gomes Maia;
Gabriane Nascimento Porcino; Cristiane de Carvalho Campos; Nayara Braga Emídio;
Maria Aparecida Juliano; Luiz Juliano; Henrique Leonel Lenzi; Paulo Marcos Zech Coelho;
Priscila de Faria Pinto; Eveline Gomes Vasconcelos;
63. INFLUENCE OF HEPATOSPLENIC SCHISTOSOMIASIS MANSONI ON THE CASTELLI’S
INDEXES
Adenor Almeida Pimenta Filho; Luiz Arthur Calheiros Leite; Bianka Santana dos Santos;
Caique Silqueira Martins da Fonseca; Ana Lucia Coutinho Domingues; Vera Lucia de
Menezes Lima
64. INVESTIGATION OF HUMAN CD59 ORTHOLOGS IN THE SCHISTOSOMA MANSONI GENOME
Cibele Aparecida Tararam; Leonardo Paiva Farias; Bogar Omar Araujo Montoya; Henrique
Krambeck Rofatto; Willian Castro-Borges; Sophie Manoel; Alan Wilson; Luciana Cezar de
Cerqueira Leite;
46
65. INVESTIGATION OF VENOM ALLERGEN LIKE PROTEINS (VALS) FROM SCHISTOSOMA
MANSONI AS VACCINE CANDIDATES
Leonardo Paiva Farias; Henrique Krambeck Roffato; Cibele Aparecida Tararam; Bogar
Omar Montoya; Rafaela Sachetto Fernandes; Patricia Aoki Miyasato; Eliana Nakano; Iain
W. Chalmers; Samirah Perally; Paul Hensbergen; Ron Hokke; Karl F. Hoffmann; Luciana
Cezar de Cerqueira Leite
66. LARGE-SCALE GENE EXPRESSION EVALUATION OF SCHISTOSOMA MANSONI ADULT
WORMS TREATED WITH MG132
Enyara Rezende Morais; Lizandra Guidi Magalhães; Érika Bueno de Carvalho Moreira;
Cláudia Sossai Soares; Katia Cristina Pereira Oliveira; Vanderlei Rodrigues
67. LOOKING FOR BIOMARKERS OF SCHISTOSOMIASIS IN A SIMPLIFIED SERUM PROTEOME
onatan Marques Campos; Leandro Xavier Neves; Milton Hércules Guerra de Andrade;
Renata Guerra de Sá; Vanderlei Rodrigues; William de Castro Borges
68. MACOPHAGE-DERIVED HEDGEHOG LIGANDS PROMOTES FIBROGENIC RESPONSES IN
Thiago de Almeida Pereira; Wing-Kin Syn; Izabela Voieta; Jiuyi Lu; Steve S. Choi; Kolade M
Agboola; Gamze F Karaca; Carlos M Antunes; William E. Secor; Zilton A Andrade; José R.
Lambertucci; Fausto Edmundo Lima Pereira; Anna Mae Diehl
69. MINING THE SCHISTOSOMA MANSONI SOLUBLE PROTEOME FOR IG-LIKE MOLECULES
Leandro Xavier Neves; Karina Taciana Santos Silva; Jonatan Marques Campos; Milton
Hércules Guerra de Andrade; Renata Guerra de Sá; Vanderlei Rodrigues; William de
Castro Borges
70. MOLECULAR MODELING STUDIES ON NOVEL INHIBITORS OF CATHEPSIN D-LIKE ENZYMES
FROM SCHISTOSOMA MANSONI
Ana Carolina Rennó Sodero; Suzanna Romero de Sousa Neves; Salvatore Giovanni De
Simone; Floriano Paes Silva-Jr
71. MOLECULAR, IMMUNOLOGICAL AND MORPHOLOGICAL ANALYSIS OF SCHISTOSOMA
MANSONI SCHISTOSOMIASIS COMBINED THERAPY WITH ARTEMISININ-PRAZIQUANTEL
Ana Júlia Pinto Fonseca Sieuve Afonso; Silvana Belo; Isabel Clemente; Ilda Jeremias;
Claudia Pen; Cátia Alexandra Costa Ferreira; Maria Amélia Grácio
72. MORPHOMETRY AND GENETIC ANALYZES OF BULINUS GLOBOSUS (GASTROPODA
PLANORBIDAE) FROM REGION OF BENGO, ANGOLA
Ângela Helena C. Velez; Maria Manuela P. Calado; Ana Júlia Afonso; Maria Amélia A.
Grácio
73. PARTIAL CHARACTERIZATION OF BIOMPHALARIA TENAGOPHILA (TAIM STRAIN)
TRANSCRIPTOME USING THE EXPRESSED SEQUENCE TAG STRATEGY – PRELIMINARY
RESULTS
Michele Araújo Pereira; Aristeu Silva-Neto; Marina de Moraes Mourão; Thomaz LücherDias; Consuelo Latorre Fortes-Dias; Paulo Marcos Zech Coelho; Glória Regina Franco
47
74. PROTEOMIC ANALYSIS OF SCHISTOSOMA MANSONI TREATED IN VITRO WITH 3METHYLCLONAZEPAM
Jean Pierre Barros Thibaut; Regina Coeli Gonçalves Lage; Vagner Simonin; Antonio
Galina; Guilherme Oliveira; François Noel
75. PROTEOMIC AND HISTOPATHOLOGICAL ANALYSIS OF PRAZINQUANTEL TREATMENT ON
LIVER OF MICE INFECTED WITH SCHISTOSOMA MANSONI
Gabriela Ayres Fragoso Nascimento; Humberto Gonçalves Bertão; Adriana Silva Andrade
Pereira; Natália Lima Cavancanti; Maria da Paz Carvalho da Silva; Mário Ribeiro de Melo
Júnior; Mônica Camelo Pessoa de Azevedo Albuquerque; Maria Elizabeth Cavalcanti
Chaves
76. PURIFICATION OF THE ENZYME URIDINE CYTIDINE KINASE OF SCHISTOSOMA MANSONI
SELECTED AS A THERAPEUTIC TARGET
Débora Meira Neris; Larissa Romanello; Alexandre Cassago; Humberto D‘Muniz Pereira;
Richard Charles Garratt; Fernanda de Freitas Anibal
77. RECOMBINANT SMNPP-5 INDUCES ANTIBODIES THAT PARTIALLY INHIBIT THE SURFACE
ENZYMATIC ACTIVITY BUT FAIL TO PROTECT AGAINST CHALLENGE WITH SCHISTOSOMA
MANSONI
Henrique Krambeck Rofatto; Leonardo Paiva Farias; Cibele Aparecida Tararam; Bogar
Omar Araujo Montoya; Robert Alan Wilson; Luciana Cezar de Cerqueira Leite
78. REDUCTION ON CONCENTRATION OF LECITHIN-CHOLESTEROL ACYLTRANSFERASE IN
PLASMA OF HEPATOSPLENIC SCHISTOSOMIASIS PATIENTS
Maciel, G.R.; Carvalho, V.C.O.; Silva, C.A; Santos, B.S. dos; Domingues, Ana Lucia Coutinho;
Lima, V.L.M
79. REFINEMENT STRUCTURE ADENOSINE KINASE AND EXPRESSION HYPOXANTHINEGUANINE PHOSPHORIBOSILTRANFERASE OF SCHISTOSOMA MANSONI
Larissa Romanello; Alexandre Cassago; Ricardo De Marco; Glaucius Oliva; Richard
Charles Garratt; Humberto D´Muniz Pereira
80. SCHISTOSOMA MANSONI ALKALINE PHOSPHATASE: DIFFERENCES IN EXPRESSION
PATTERNS ALONG LIFE CYCLE IN THE CONTEXT OF VACCINE DEVELOPMENT
Bogar Omar Araujo Montoya; Cibele Aparecida Tararam; Henrique Krambeck Rofatto;
Leonardo Paiva Farias; Luciana C.C. Leite; R. Alan Wilson
81. SCHISTOSOMA MANSONI GENOMIC AND FUNCTIONAL GENOMICS DATA INTEGRATION:
SCHISTODB.NET
Guilherme C. Oliveira; Roney S. Coimbra; Jessica Kissinger; Fabiano Mirsky Pais; Mariana
C. Simões; Adhemar Zerlotini;
82. SCHISTOSOMA MANSONI VACCINE CANDIDATE ANTIGEN SCREENING USING PROTEOMIC
TOOLS
Fernanda Ludolf Ribeiro; Rosiane A. da Silva-Pereira; Paola Patrocínio; Andréa Gazzinelli
Corrêa de Oliveira; Rodrigo Corrêa Oliveira; Guilherme Corrêa Oliveira
48
83. STRUCTURE OF METHYLTHIOADENOSINE PHOSPHORYLASE (MTAP) FROM SCHISTOSOMA
MANSONI
Juliana Roberta Torini de Souza; Alexandre Cassago; Ricardo De Marco; Glaucius Oliva;
Richard Charles Garratt; Humberto D‘Muniz Pereira
84. THE DETECTION OF SCHISTOSOMA MANSONI TRANSRENAL DNA IN URINE SAMPLES
USING THE MURINE MODEL
Guilherme de Oliveira e Silva; Martin J. Enk; Nilton Barnabe Rodrigues
85. THE ROLE OF IRON IN THE MODULATION OF GRANULOMATOUS RESPONSE IN MURINE
SCHISTOSOMIASIS MANSONI MODEL
Alice Maria de Magalhães Ornelas; Flávia Rachel Moreira Lamarão; Guilherme de
Bustamante Pereira de Miranda; Bernardo Miguel de Oliveira Pascarelli; Marcelo Ribeiro
Alves; Milton Ozório Moraes; Marcelo Pelajo Machado
86. SCHISTOSOMIASIS CONTROL PROGRAM IN ARACAJU/SE: PROGRESS AND PROBLEMS
Nathalia Vasconcelos Barroso, Aloisio Ferreira Pinto Neto, Robergson Rozendo Ribeiro,
Andréa Valença Cardoso, Luciene Barbosa, Karina Conceição Gomes Machado de Araujo,
Satie Katagiri, Roseli La Corte dos Santos
49
50
nEW StRatEgIES Of SCHIStOSOMIaSIS COntROL and SURvEILLanCE In SãO PaULO
(BRazIL).
Horacio Manuel Santana teles
Secretaria Estadual de Saúde de São Paulo, São Paulo, Brasil
In Sao Paulo, the detection of the first autochthonous cases of schistosomiasis in Santos
happened in the early 20th century past. Already in the mid-50s, followed by the discoveries
of new cases and outbreaks of disease in the valley of the Paraiba do Sul Subsequently,
the findings of cases and outbreaks have indicated that disease transmission occurred in
endemic areas is more extensive.
Confirmed the transmission of Schistosoma mansoni was followed by a few isolated attempts
to control, until in 1960 the issue merited a plan to combat the snail intermediate host with
molluscicides. In 1968, there was the perceived need to organize the actions of the program
focusing on reversing the transmitting species of snails (Biomphalaria glabrata and B.
tenagophila), but now including diagnosis and treatment of cases, carrying out small works,
such as, landfills, pipelines and cleaning of hydric environments.
Following in 1978, a fact worthy of note was the inclusion of schistosomiasis in the list
of notifiable diseases. The guidelines expressed in the manual, also gave precision to the
laboratory procedures, considering the age and origin of those addressed in the public
health system. Refinements for the examinations of stool samples provided the improved
resolution of the processing of cases. On occasion, the advent of oxamniquine favored the
goal of reducing morbidity and prevalence of the disease, with the expectation of further
reducing the number of parasite eggs eliminated in the environment. These guidelines
remain valid until the end of the 90s. With effective treatment, the continuity of work and
gradual improvement of basic sanitation almost eliminated the possibility of developing
severe disease. The decline in notifications of the average of 20 000 cases/year of the 80 to
10 000 cases/year in the next, and for two thousand cases per year since 2001, with only 23
municipalities whose notifications 100 cases/year exceed, and only nine demonstrated the
occurrence of naturally infected snails.
In 2004, for policy determination, coordination of surveillance actions rose to the Divisão de
Doenças de Veiculação Hídrica e Alimentar (DDVHA) of the Health Secretarial of State. While
the program proposed in 2006 not incorporate significant changes in the foundations of
control, from 2008 established the goal of eliminating autochthony with the intensification
of control in the pre-established with the support of sentinel units to the rear for laboratory
confirmation of cases considered as suspects by the units of health care districts situated in
areas at risk of acquiring the disease. The practice of the new model came in 2009 and 2010,
with the realization, respectively, of the 1st and 2nd Week of Schistosomiasis. Given the
dominance of asymptomatic or mild clinical manifestations, nonspecific, with no complaints
and therefore no reasons to suspect the existence of the disease by the population and by
health professionals, besides the aforementioned goals, work intended to raise awareness
of assistance doctor to the problem. Presumption in the case, is also proposed to take blood
samples with additional serological techniques more refined and sensitive, which in positive
cases, resulted in the repeated examination of stool samples for confirmation and treatment
of infections. Available information shows that work motivated the deployment of about 200
51
sentinel units at the prospect of attendance of at least 20 000 people per year.
In the new model, the partial results show the discovery in 2009 of 1245 cases, of which 181
(14.5%) native. In 2010, until April, with only the routine work, 162 cases were diagnosed,
with 28 (17%) autochthonous, coming from all municipalities in endemic areas. The results
of weeks of work this year are not yet available.
Apart from the discussion of whether or not the new model immediately, it is anticipated
the need to overcome some obstacles of a purely political, in the absence of changes in
prophylactic procedures designed to control. The main problems are directly related to the
need for timeliness and coverage of the health network to the problem of collecting and
treating sewage, particularly in endemic areas, and especially the integration of different
bodies with responsibilities for the development of action prophylactic, things that depend
primarily on the political will of governments and leaders.
In addition, the assumption of a successful target remains tied to the social and economic
standing of the population which permits the reduction of disparities in living conditions and
housing the population of the state.
52
EnvIROnMEntaL and SOCIOCULtURaL aSPECtS Of SCHIStOSOMIaSIS tRanSMISSIOn:
tHE CaSE Of PORtO dE gaLInHaS, PERnaMBUCO.
Centro de PesquisasAggeu Magalhães, Fiocruz, Pernambuco
In order to understand the occurrence and causality of endemic diseases like schistosomiasis,
it has to be understood that although the disease is manifested in individuals, the health
situation is a manifestation of human activities within the environment, and that maintenance
of the disease depends on a spatial-temporal structure that is historically, socially and
culturally constructed. The dimensions of the determining or causal factors can only be
understood through multidisciplinary approaches that start from the principle that there is
interdependence between individuals and the biological, social and environmental contexts
external to these individuals. This interdependence influences individuals’ wellbeing and/
or promotes threat to their health. In this respect, the importance of local baseline studies
for understanding health hazards can be highlighted. Epidemiological analysis tools can be
included in such studies to elucidate the environmental, ecological and behavioral issues
involved in the dynamics of endemic disease transmission. These studies have successfully
used geoprocessing tools for spatial analysis on health hazards and have sought to evaluate
the impact of adverse environmental conditions on individuals’ health and integrity within
time and space. Geoprocessing consists of a set of techniques for gathering and dealing
with spatially organized information, using Geographic Information System (GIS) tools
and the Global Positioning System (GPS), with the aim of developing adequate models
for monitoring, predicting and preventing risks. These techniques contribute towards
etiological exploration of health events, with emphasis on spatial descriptions of events,
identification of environmental and occupational risks and analysis of health situations in a
given geographical area, thereby facilitating comprehension of the phenomena in their full
complexity. In Pernambuco, schistosomiasis is becoming endemic in coastal localities where
disorderly urban growth has brought visible environmental impacts, consisting of devastation
and pollution of the natural environment. Less favored social groups coming from rural areas
that are endemic for schistosomiasis congregate in peripheral areas without any housing
and sanitation infrastructure, thereby creating devastated and unhealthy environments. This
chaotic occupation has led to loss of the characteristics of the district of Vila de Porto de
Galinhas, with increased population density in central areas and indiscriminate landfill in
mangrove swamp areas, where groups of shantytown dwellers gather. They live in situations
of underemployment, doing casual work generated by the main local economic activity:
tourism. It is estimated that an average of 6,000 tourists visit the locality per month, coming
either from the local region or from other states and countries. The districts of Merepe, Vila
de Porto de Galinhas, Salinas, Pantanal and Socó present a contingent of fixed population,
represented by natives of the area, and a floating population composed of service providers
involved in local tourism. Promotion of tourism provides an economically profitable alternative
that generates employment and income for the population, but collective action at different
levels of public administration (municipal, state and federal) is needed in order to define
a development model that ensures sustainability of the environment and improvements
in quality of life for the local population. In the year 2000, an outbreak of schistosomiasis
53
was recorded in Porto de Galinhas, Pernambuco, when natural environmental events
(heavy rain and flooding), along with human actions that disturbed the ecosystem (landfill,
introduction of vector snails, ditches with leakage and streets without drainages) were
responsible for 663 acute cases of schistosomiasis among local residents and vacationers.
The epidemiological information generated by this event was put into a spatially organized
form and this emphasized the importance of aggregating the biological data and geographic/
environmental data, in order to understand the ecological context within which the health
hazard occurred. So far, few environmental or sanitary improvements have been made in the
district of Porto de Galinhas aimed at diminishing the transmission of schistosomiasis. Even
today, infected snails can be found in the streets, this indicating that there is a risk of new
outbreaks of the disease if new floods occur. Since July 2010, a new project has been under
development in this locality, with the aim of updating the epidemiological and environmental
situation and comparing it with data from the last ten years. The epistemological approach
in this study is the concept of environmental risk, which methodologically involves working
with space and health, environment and risk, and environmental indicator construction. This
study can be justified in the light of the need to inform and provide tools for local healthcare
services, so that the required care can be provided through the surveillance and monitoring
actions, thereby preventing occurrences of new outbreaks of schistosomiasis and improving
the community’s quality of life.
54
SCHIStOSOMIaSIS tRanSMISSIOn COntROL tHROUgH
CHEMOtHERaPY and SanItatIOn IS a REaL POSSIBILItY
aSSOCIatIOn
Of
naftale Katz
Centro de Pesquisas René Rachou/FIOCRUZ and Academia Mineira de Medicina, Belo
Horizonte, Brazil
Two endemic areas of schistosomiasis mansoni were studied for about 25 years. In the first
one, Comercinho, at the Northeastern part of the State of Minas Gerais, the first clinicalepidemiological survey was done in 1974. In 1981, the staff went back and reexamined the
total living population of the city. The patients eliminating Schistosoma mansoni eggs in their
feces were treated with oxamniquine. Same procedures (exams and treatment) were repeated
in 1981, 1986, 1988, and in 1992 praziquantel has been introduced. The results from the last
survey (2005) were compared with those obtained in 1981. From a population of 1474 people
studied in 1981, 475 individuals could be found and 358 out of them were submitted to stool
examination, and 231 were clinically examined. The comparison of the results of 1981 and
2005 clearly demonstrated that transmission control was obtained, although not completely.
In fact, prevalence in Comercinho decreased significantly from 70.4 to 1.7%, as well as the
presence of hepatointestinal form (25.3 to 3.5%), hepatosplenic form (6.8 to 1.3%). No one
single new case of hepatosplenic form developed in more than 20 years of follow-up.
In the last survey in Comercinho, 96% of the houses disposed of safe water supply by means
of the public system, 97.6% had closet-bowls or cess-pits for waste disposal and 97.6% were
classified as being of better quality. In the 1981 survey, the data were 33.7, 71.7 and 34.2%,
respectively. In the second one, Ravena, district of Sabará, State of Minas Gerais, the control
program started in 1980. Initially, the prevalence of schistosomiasis was 36.7%. No cases of
hepatosplenic form were found. A specific treatment with oxamniquine in large scale was done
(every four years, three treatments) to patients eliminating eggs in their stools. Domestic water
supply was provided, to 90% of the housings, and appropriate waste disposal was present in
only 17% of the houses. In 1992, the prevalence in the population decreased to 11.5% and
appropriate waste disposal increased to 36%. From 1992 onwards, the population was treated
by a physician at the local Health Center, based on the results of stool examinations and by
spontaneous plea.
In 2008, this area was re-examined, i.e., 27 years after the first clinical-epidemiological survey.
The prevalence was 2.5% (a decrease of approximately 95% compared with that found at 1980).
Besides, 25% of the houses disposed of safe water supply, and more than 80% had appropriate
waste disposal. Interestingly, the majority of the population is no more in the habit of using
natural contaminated water. Currently, in both areas, the lack of low rate of infected snails, low
schistosomiasis prevalence rate, absence of new cases of hepatosplenic form, decrease of liver
enlargement, have clearly proved that control measures in association, chemotherapy, water
supply, and waste disposal can led to interruption or significant decrease of transmission and
to morbidity control. Finally, due to the effectiveness of those measures used, the Brazilian
Government is urged to adapt this association in order to obtain schistosomiasis control in the
country.
55
RURaL ELECtRIfICatIOn In BRazIL, IMPLICatIOnS fOR SCHIStOSOMIaSIS
tRanSMISSIOn: a StUdY Of a RURaL COMMUnItY In MInaS gERaIS StatE
Helmut Kloos
Department of Epidemiology and Biostatistics, University of California Medical Center, San
Francisco, USA
Introduction: The Brazilian government implemented in 2003 a rural electrification program
(Luz para Todos-Light for All) to provide complete electricity coverage for all citizens, with
the aim of promoting socioeconomic development and health and reduce inequalities. This
implies the provision of electricity to 15 million people, mostly in rural communities, by the
end of 2010. By 2009, about 11 million people had benefited from this program. A number of
studies world-wide have described the rapid increase in energy consumption and associated
improvements in household welfare; health and infrastructure but few studies have
examined unintended health risks. This study briefly summarizes the rural electrification
program in Brazil and presents information on potential schistosomiasis exposure risk
conferred by the installation of electric pumps in shallow wells in Virgim das Graças rural
area in the northern part of Minas Gerais State. Methodology: All complying members (more
than 95%) of the 145 households in Virgim das Graças in 2001 and of the 144 households
in 2009 were interviewed about their water supply, water use patterns, and socioeconomic
status using a questionnaire. The domestic water supply consisted predominantly of
piped connections in the central village, and of piped springs, shallow wells (cisternas and
cacimbas), and streams in the rural area. Residents were examined for S. mansoni infections
in 2001, 2002, 2005 and 2009 using the Kato-Katz method and treated with praziquantel. All
potential snail habitats in the study area were surveyed for intermediate hosts and infections
3 times during the dry and wet seasons in 2001 and 2002. Six electric pumps that had been
installed in wells were visited and examined for snails in 2010. These various activities were
carried out as part of a longitudinal and interdisciplinary epidemiological study. Results:
The proportion of households with electricity, piped spring water, water tanks, and piped
well water increased sharply between 2001 and 2009 and the number of persons admitting
that they regularly come in contact with stream water decreased during that 8 year period.
All 38 households with installed electric pumps in 2009 pumped well water through plastic
pipes to storage tanks supplying bathrooms, kitchens and sites adjacent to the houses for
bathing, cooking, washing utensils and other household uses. Sixteen of these households
shared pumped water with neighbors, 12 of them sharing water with a single neighboring
household, 1 household sharing water with another two households and 3 households
had reciprocal sharing arrangements with other neighbors. Three of 6 wells visited in 2006
contained Biomphalaria glabrata. Another 3 wells had yielded S. mansoni-infected snails
during snail surveys in 2001/2002. S. mansoni infection rates and geometric mean egg counts
in 2009 were not significantly different in the households pumping water from wells than
those not owning a pump (P<.05) but were 18.7% lower among households receiving safe
piped water from the protected source in the village (24.6%) than in the rural households
(29.3%). Conclusions: The recent installation of a relatively large number of electric pumps
in potential snail habitats, widespread sharing of well water with neighbors, and recovery
of S. mansoni infected B. glabrata from some of these sites in Virgim das Graças indicates
56
both a high demand for pumped well water in the study area and the potential spread of
schistosomiasis via pipes to households This situation further suggests that other rural
communities receiving electricity through the rural electrification program may also be
at risk of infection from piped water in the home but additional studies are required in
other schistosomiasis-endemic areas to determine the extent of the problem. If necessary,
information needs to be provided by the water authorities to local populations on the health
risk of installation of pumps in shallow wells and other domestic water sources serving as
potential habitats of snail intermediate host snails to promote awareness in the population
and reduce the risk of S. mansoni transmission at these sites.
Financial support: CNPQ, FAPEMIG, NIH-ICIDR Grant 1R03AI071057-01,CAPES.
57
fIBROSIS and SCHIStOSOMIaSIS
zilton andrade
Centro de Pesquisa Gonçalo Moniz, Fiocruz, Salvador, Bahia
Fibrosis is an outstanding feature of the pathology of schistosomiasis. It starts from the
miracidial secretions which are eliminated trough the micro-pores in the egg shell. This
elimination is an essential step to facilitate egg extrusion through the host intestinal wall
toward the intestinal lumen. But, when the eggs are instead retained within the host
tissues, such secretions, which contain lytic and antigenic materials, induce focal chronic
inflammation, with angiogenesis and fibrosis.
Several factors may influence the clinical significance of fibrosis in schistosomiasis, the main
one of them being the parasite load. But, host age, nutrition, genetic background, abnormal
or unusual location of worms, etc may play a role. The host production and regression of
fibrosis in general, as well as in schistosomiasis, are under a dynamic equilibrium: when the
fibrogenic factors predominate there is increased synthesis of extracellular matrix, especially
of types I and III collagens and of their associated proteins; when the opposite occurs there is
increased activity of collagenolytic factors (collagenases) and remodeling. Interestly enough,
recent findings have indicate that angiogenesis may than play a dual and paradoxical role
during fibrosis production as well as during fibrosis regression. The clinical significance
of schistosomal fibrosis is quite obvious when it is associated with the pseudo-neoplastic
forms, the neural involvement, but is not so when one considers the hepatoeplenic form
of the disease or its cardio-pulmonary complication. In these two examples the vascular
obstructive changes determine the clinical course, not fibrosis itself. The classical anatomic
picture of pipe-stem fibrosis with splenomegaly may be observed in asymptomatic patients,
and there is no direct correlation between schistosomal hepatic fibrosis and the severity of
its clinical manifestations.
58
nEUROSHIStOSOMIaSIS
tereza Cristina a. ferrari
Departamento de Clínica Médica, Faculdade de Medicina, Universidade Federal de Minas
Gerais, Belo Horizonte, MG, Brasil.
Introduction: The involvement of the central nervous system (CNS) by schistosomes may or
may not cause clinical manifestations. When symptomatic, neuroschistosomiasis (NS) is one
of the most severe presentations of the infection whose prognosis is largely dependent on
early recognition and treatment. Schistosoma mansoni and S. haematobium cause preferably
myeloradiculopathy, and S. japonicum almost always encephalic disease. The neurological
manifestations are due to numerous eggs and granulomas grouped in confined areas of the
spinal cord or the encephalus. Although knowledge on NS has improved during the last two
decades, several aspects of this disorder are unknown and deserve investigation. During a
period of 22 years, we have developed investigations on the different aspects of spinal cord
schistosomiais mansoni (SCS). These studies are briefly described below. Methodology:
We evaluated and followed up 72 patients with SCS admitted to Hospital das Clínicas,
Universidade Federal de Minas Gerais, Belo Horizontal, MG, Brazil. These patients were
investigated according to a specific protocol, treated with corticosteroid and praziquantel,
and followed up for a long time. Based on these cases, we studied the following aspects of SCS:
1) the clinical features and results of routine laboratorial tests; 2) the value of cerebrospinal
fluid (CSF) IgG against soluble egg antigen of S. mansoni (anti SEA IgG), measured by ELISA,
as a diagnostic tool for SCS; 3) the profile of immune response of the disease, investigated
by the measurement of cytokines in paired serum and CSF samples from SCS patients and
health controls; and d) the intrathecal synthesis of IgG by the estimation of the CSF IgG index.
Additionally, the clinical and laboratorial aspects of three patients with cerebral schistosomiais
mansoni admitted to our institution were analyzed along with dada from the reported cases
of this form of NS. Results: SCS usually presented as a low cord syndrome of acute/subacute
progression frequently associated with the involvement of the cauda equine roots. Lower
limbs pain, weakness and sensory disturbance, and autonomic dysfunctions, particularly
bladder dysfunction, were often present. Gastrointestinal and other manifestations of the
schistosomal infection were commonly absent. CSF examination showed an inflammatory
pattern with a mild/moderate increase in both total protein concentration and lymphocyte
count. Eosinophils were observed in the CSF of about 50% of the cases. Magnetic resonance
imaging demonstrated signs of inflammatory myelopathy. Stool parasitological examination
failed in revealing S. mansoni eggs in a large proportion of the patients; and, schistosome egg
counts, performed in rectal biopsy of these individuals, indicated a low parasite burden in the
majority of them. The SCS patients usually presented response to the treatment; however,
only about 60% of them fully recovered or persisted with minimum neurological deficit
compatible with an unrestricted quality of life. The remaining individuals were left with some
limiting deficit. Anti SEA IgG was detected in the CSF of the majority of the patients with SCS.
CSF concentrations of anti SEA IgG below 0.10microg/mL practically exclude the diagnosis
of the disease, and values higher than 1.40microg/mL confirm its diagnosis with reasonable
precision. The comparison of the levels of the cytokines between CSF and serum from the
SCS patients and the control groups, and between CSF and serum of the individuals with
59
SCS was compatible with an inflammatory as well as a skewed type-2 immune response that
probably occur both systemically and within the CNS in SCS. Intrathecal synthesis of IgG was
demonstrated in 80% of the SCS patients. Cerebral NS usually presents as a slow-expanding
intracranial tumor-like lesion. Its clinical manifestations are variable and depend on the
increased intracranial pressure and on the site of the lesion. Headache, seizures, visual
abnormalities, speech disturbances, sensory impairment, hemiparesis, nystagmus, vertigo
and ataxia are common manifestations. Antischistosomal drugs, steroids and surgery are
the therapeutic modalities currently available for treating this form of shcistosomiasis. Both
surgical resection and medical therapy have been used successfully. Cerebral NS is associated
with a better outcome than the spinal cord disorder. Conclusion: Spinal cord and cerebral
NS are severe conditions. A high index of suspicion is necessary in order to diagnose and
treat these disorders promptly as their outcome is largely dependent on early treatment. We
continue studying NS since several its aspects are unknown or unclear.
Financial support: CAPES, CNPq, and Fapemig.
60
SCHIStOSOMaL gLOMERULOPatHY
Washington LC dos-Santos
Laboratório de Patologia e Bio-Intervenção, Centro de Pesquisas Gonçalo Moniz, Fiocruz,
Salvador, Bahia.
The occurrence of glomerulpathy in patients with schistomiasis was recognized and
characterized in a series of works published, mainly by Brazilian researchers, in the 60s and
70s. Soon it became clear the association of different patterns of glomerular lesions with the
hepatosplenic form of the schistosomiais. Membranoproliferative glomerulonephritis and
Focal segmental glomerulosclerosis emerged as the most prevalent form of renal lesion in
these patients. The suggested mechanism underlining Schistosomal glomerulonephritis was
immune complex deposition. Immune complexes of IgG, IgM and complement, together with
parasite antigens and also IgA aggregates have been identified in glomeruli of both in human
and experimental animals, at different stages of the disease. Immune complex deposition may
be intensified by the portosystemic shunt secondary to portal hypertension. Co-infection by
Salmonella added an exudative characteristic to the glomerular lesions. Although immune
complex deposition has been consistently demonstrated in schistosomal glomerulonephritis,
the mechanism associating Focal segmental glomerulosclerosis with schistosomiasis remains
unknown. The genetic background of a predominantly afro-descendent population infected by
the parasite has been proposed as a possible explanation. The diversity of glomerular lesions
associated with schistosomiasis motivated a clinicopathologic classification of schistosomal
glornerulopathies, proposed in 1992 by the African Association of Nephrology. Such
classification recognizes four categories of glomerular lesions: class I (Mesangioproliferative
glomerulonephritis), class II (Exudative glomerulonephritis), class III (Membranoproliferative
type I or type III glomerulonephritis); class IV (Focal segmental glomerulosclerosis), and class
V (Amyloidosis). Recently, another class VI of lesion has been proposed, consisting of an
association of mesangial proliferation, apoptotic transit cellular infiltrations, amyloidosis, and
cryoglobulin deposition, attributed to the combined effect of infections by Schistosoma and
by the hepatitis C virus. Schistosomal glomerulpathy is more frequently associated with S.
mansoni and S. japonicum infections. In Brazil mass treatment with oral drugs has lead to a
decrease in the occurrence of the hepatosplenic form of schistosomiais with similar impact
in the occurrence of schistosomal glomerulpathy. A recent survey carried out in the Service of
Renal Pathology-CPqGM-FIOCRUZ, revealed a decline in the mention of S. mansoni infection in
biopsy request forms, from 16% to 2% in the period of 2003 to 2009. Positive test for S. mansoni
infection was reported in 24/689 (4%) patients. In one of these patients, hepatosplenomegaly
was also reported. Among the whole group of patients with a positive tests for S. mansoni
infection 7/24 had Focal segmental glomerulosclerosis and 4/24 had Membranoproliferative
type I glomerulonephritis (including the patient with reported hepatosplenomegaly). It is
interesting to note during the studied period of 2003 to 2009, that the average frequency of
membranoproliferative glomerulonephritis remained stable, comprising 5% of the biopsies.
However, the most commonly associated diseases with this pattern of glomerulonephritis in
the period were Systemic Lupus Erythematosus and Lymphoproliferative diseases.
Financial support: FAPESB, CNPq
61
ULtRaSOUnd vERSUS BIOLOgICaL MaRKERS In tHE dIagnOSIS Of PERIPORtaL
fIBROSIS
ana Lúcia Coutinho domingues
Universidade Federal de Pernambuco
The assessment of liver periportal fibrosis (PPF) provides useful information for the diagnosis
and for therapeutic decision in patients with Schistosoma mansoni and is an important aproach
in the fields studies. Although the liver wedge biopsy is the most accurate method for the
fibrosis assessment it has some limitations and risks, and the needle biopsy doesn’t have a good
sensibility for the diagnosis of PPF. Actually upper abdominal ultrasound became the most used
diagnostic tool to diagnose and quantify the PPF, but it is not available in all endemic areas,
because it require US equipment and qualified examiners. This has led to the development
of non invasive bioquemical markers of liver fibrosis.The characteristics of an ideal marker of
liver fibrosis could be non invasive, liver specific, easy to perform, mensurable by sensitive,
reproducible and fast methodology. From the pratical point of view the aim of biochemical non
invasive investigation in mansoni schistosomiasisis is to discriminate between patients with no
from mild fibrosis and with mild from advanced fibrosis. The accuracy of a test is given as the
area under the curve (AUC) of the receiver operator characteristic (ROC). An ideal marker would
have na AUC of 1.0 and them a 100% sensibility and specificity. The aim of this talk is to review
ours and others studies that correlate serum markers of hepatic fibrosis with the diferents
pattern of PPF established by ultrasound using the Niamey Protocol in mansoni schistosomiasis.
We will focused in hiarulonic acid (HA), platelets, immunoglobulin G (IgG), and APRI (ratio AST/
platelets) índex. Hialuronic acid We have studied 122 patients with PPT and 12 schistosomotic
patients without no fibrosis. Serum levels of HA acid in no fibrosis group was 23.9 mcg/l, mild
fibrosis was 51.9 mcg/l, and 64.3 mcg/l those with advanced fibrosis. To indentify the best cutoff
for patients with fibrosis the ROC plot showed a serum value of 27.8 mcg/ with sensibility of
78.2% and specifity of 83,7%. In a previous study in 2007 with 51 we found a serum HA level
of 20,2mcg/l to diferenciate patients with milder PPF from those with more severe PPF with
a sensitivity of 60% and a specifity of 65%. Köpke-Aguiar et al, found a level of 20.0 mcg/l of
HA to separate shistosomiasis patients with and without portal hypertension. Platelet count
(PLt) and aPRI index –Combined assessment of AST/PTL ratio (APRI) had a high diagnostic
value for cirrhosis and has been used in schistosomotic patients. In our study with 122 patients
platelet count had significant negative correlation to the different periportal fibrosis patterns
which means that the more advanced stages of fibrosis had lower platelets count as seen in
no fibrosis group (257.833 ± 74.215) mm3 ,mild fibrosis (158.355 ± 79.729) mm3, and severe
(96.430 ± 45.355) mm3. The platelet count accuracy, represented by area under curve “ROC”,
to identify patients with periportal fibrosis was 0,921, being the value of 171.000 mm3 the best
cutoff with 80% of sensibility and specificity of 91.7%. APRI index showed progressive values
according to the groups: 1 (0,26 ± 0,11), 2 (0,92 ± 0,98) and 3 (1,58 ± 1,40) and accuracy to
identify fibrosis was 0,930 , and the value of 0,349 as the best cutoff with sensibility of 90% e
specificity of 83,3%. The group of Lambertucci had studied platelets and APRI with good results.
Immunoglobulin g (Igg)- Recent studies have found that immunoglobulins exert a direct effect
on hepatic fibrogenesis and that IgG stimulating the proliferation of HSC. Recently our group in
41 patients showed na increased of the IgG serum levels according to the progression prom PPF
62
intensity. In our actual study was obseved that the medium serum levels of IgG were 1236 mg/dl
(± 368) in group 1, 1338 mg/dl (± 379) in group 2 and 1647mg/dl (± 242) in group 3. The value of
1542mg/dl was the best cutoff to identify those with advanced fibrosis with sensibility of 57,3%,
specificity 91,7% and accuracy of 0,736. Conclusion: platelets, IgG, HA and APRI índex had
correlation with periportal fibrosis diagnosed by ultrasound and are promising serum markers
of fibrosis in mansonic schistosomiasis. But Platelets and APRI index were the best predictors
of fibrosis when used singly and like they are easier to maker in the endemic areas, they can
be used as a selection of patients who have most advanced desease and will need to make an
ultrasound and more investigations.
63
MagnEtIC RESOnanCE In tHE dIagnOSIS Of HEPatOSPLEnIC SCHIStOSOMIaSIS
José Roberto Lambertucci
Departamento de Clínica Médica – UFMG, Belo Horizonte, MG
Introduction: Magnetic resonance imaging (MRI) is primarily a noninvasive medical imaging
technique used in radiology to visualize detailed internal structures and limited function of
the body. MRI provides much greater contrast between the different soft tissues of the body
than computed tomography (CT) does. Magnetic resonance angiography (MRA) generates
pictures of the vessels to evaluate them for stenosis or aneurysms. One advantage of an MRI
scan is that it is believed to be harmless to the patient. It uses strong magnetic fields and
non-ionizing radiation in the radio frequency range, unlike CT scans and traditional X-rays,
which both use ionizing radiation. Contrast agents may be injected intravenously to enhance
the appearance of blood vessels, tumors or inflammation. Methodology: In the present
review, the role of MRI in the evaluation of morbidity in schistosomiasis, including liver
fibrosis/portal hypertension , is emphasized. Manson’s schistosomiasis causes periportal
fibrosis (or Symmers’ white clay pipe-stem fibrosis) and portal hypertension in about 6%
of infected subjects, usually with preservation of the hepatic function. The assessment of
liver involvement is of major importance in determining prognosis and risk of complications
from schistosomiasis such as upper digestive bleeding secondary to variceal rupture.
Results: For many years the diagnosis of hepatosplenic schistosomiasis and liver fibrosis was
confirmed by abdominal palpation and the identification of liver and/or spleen enlargement.
However, there is no consensus about the clinical parameters of the liver and spleen to be
considered in the evaluation. Hepatomegaly and splenomegaly have no uniform definition.
The limit for the determination of an increase in liver volume remains a disputable issue.
Some investigators consider hepatomegaly to be present when the organ exceeds the costal
margin by 5cm along the sternal line or by 4cm along the midclavicular line. Others accept
as enlarged a liver palpable 2cm below the costal margin or simply a palpable liver, without
mentioning if the organ is palpable at rest or during inspiration. Still, other investigators
attribute more importance to liver characteristics such as a hard consistency, the presence of
nodules on the hepatic surface, or the prominence of the left lobe. Regarding the spleen, its
palpability would be sufficient for some physicians, whereas others believe that the spleen
should be palpable at the costal border or beyond it. Thus, the diagnosis of hepatosplenic
schistosomiasis varies according to the different criteria mentioned. For the last two decades,
abdominal ultrasound (US) has become the best imaging technique to evaluate liver fibrosis
caused by schistosomiasis mansoni. It is an indirect method for diagnosing and classifying the
disease. Ultrasonographic examination of subjects with hepatosplenic schistosomiasis has
detected a characteristic pattern of abnormalities, quite different from the aspects observed
in liver cirrhosis and in acute schistosomiasis. The most important findings are echogenic
thickening of the walls of the portal vein and its branches, indicative of periportal fibrosis,
and echogenic enlargement of the gallbladder wall. However, US is a subjective procedure
and thus examiner-dependent. Magnetic resonance imaging (MRI), differently from US,
is less examiner-dependent. MRI findings have added a series of precious information to
ultrasound and clinical examination, such as: a clear image of liver fibrosis easily identified
by inexperienced eyes and a nice picture of the spleen and kidney; the abdominal vessels are
64
also well defined pointing out the collateral veins and the presence/absence of portal vein
thrombosis. Conclusion: In brief, there is no definite algorithm for evaluating schistosomiasisrelated morbidity. The association of a comprehensive history and physical examination,
basic laboratory tests (stool examination for S. mansoni eggs and blood cell count), and
imaging methods seem to offer the best approach to evaluating patients with this disease.
For the time being, MRI’s costs are restricting, which prevent it from being used in fieldbased studies or routinely in developing countries. However, in situations where research is
involved or in patients with severe disease for whom surgery for portal hypertension is being
considered MRI seems to be a wise choice.
65
MOLECULaR dIagnOSIS Of SCHIStOSOMIaSIS
Martin Johannes Enk, bLuciana Inácia gomes, c,dnilton Barnabé Rodrigues, cguilherme
Oliveira e Silva, bana Rabello, aPaulo Marcos zech Coelho
a
Laboratório de Esquistossomose, Centro de Pesquisas René Rachou, Fiocruz, Belo Horizonte,
Brasil; bLaboratório de Pesquisas Clínicas, Centro de Pesquisas René Rachou, Fiocruz, Belo
Horizonte, Brasil; cLaboratório de Imunologia Celular e Molecular, Centro de Pesquisas
René Rachou, Fiocruz, Belo Horizonte, Brasil; dLaboratório de Pesquisas Clínicas - Escola de
Farmácia, Universidade Federal de Ouro Preto (UFOP), Ouro Preto, Brasil
a
Introduction: Routinely the diagnosis of infections with Schistosoma mansoni is based on
the detection of parasite eggs in stool. This approach is inexpensive and easy to perform,
and provides basic information on prevalence and infection intensity. However, a limitation
of these coproscopic methods is their lack of sensitivity, especially in low endemic areas
and among individual infections with low parasite load. Besides this intrinsic limitation of
coproscopic stool examinations, the positive effect of successful control programs as well as
the rising numbers of infected travelers makes the development of more sensitive diagnostic
methods urgently needed. In this context the application of PCR-based techniques is a
promising approach, as they are scientifically recognized for being absolutely specific and
highly sensitive, relying on the direct detection of S. mansoni DNA from samples of human
feces, serum or urine. In the present study detection rates for two molecular techniques,
one using feces and the other one urine, were assessed among a population in a lowtransmission area and compared to parasitological methods. Methodology: Out of the 214
registered inhabitants of a semi-urban community, Pedra Preta in the state of Minas Gerais,
Brazil., 194 were included into this survey (20 were excluded because they did not provide
the required stool or urine samples). The participants were asked to provide a total number
of four stool samples collected on consecutive days. Twelve Kato Katz (KK) smears were
prepared from the first stool sample and two smears from the second, the third and the
forth, respectively, resulting in 18 slides for each participant. The cumulative results of all KK
slides per participant combined with two other parasitological methods, namely the Saline
Gradient technique and the Miracidia Hatch test, served as reference value for comparisons
with both molecular techniques. Additionally 0,5 gram of the first stool sample was collected
for examination according to the PCR technique described by Pontes. A urine sample of 10ml
of each participant was used for the validation of a novel DNA extraction method, which
is presented in depth in another session during this symposium, followed by amplification
with the same primers as for the stool samples, targeting the 121 bp tandem repeat
DNA sequence of S. mansoni. Results: Among the 194 participants a total number of 69
positives (35.8%) were detected by the combination of the three parasitological techniques.
Examining 18 KK smears of four stool samples resulted in 64 positives (33.0%), which stays in
strong contrast to the number of 22 infected (11.3%) revealed by the examination of a single
KK slide. In case of the PCR analysis of stool samples 62 positives (31.4%) were detected
of which 10 could not be confirmed by the parasitological techniques and 18 positives
according to the parasitological techniques could not be identified by the molecular method.
The examination of the urine samples applying the PCR technique revealed 114 (58.8%)
infected with schistosomiasis. It is worth to note here that all 69 positives detected by the
66
parasitological techniques were found by the PCR analyses of the urine samples, and 4 of the
45 positives without parasitological confirmation were also detected by the PCR analysis of
stool samples.
Conclusion: In summary the data demonstrate the future potential of these molecular
techniques for the detection of infections with S. mansoni, considering the high detection
rates achieved by a single examination. The discordant results between Kato-Katz analysis
based on twelve slides and the Schistosoma PCR examining the same amount of stool of the
first sample can be explained that 14 out of 24 (58.3%) were positive in subsequent Kato-Katz
examinations of additional samples, indicating that the PCR method is more sensitive than
the Kato-Katz technique. The same applies, up to a certain extend, for the PCR method using
urine samples. Nevertheless the assessment of the PCR in urine is more complex, due to the
fact that transrenal DNA, which reflects cellfree DNA filtered by the kidneys, is amplified.
Up to now it is not clear how long cellfree and consequently transrenal Schistosoma DNA
remain detectable after the death of the parasite, which without any doubt impacts on the
differentiation between current and past infections. As the detection of transrenal DNA does
not depend on the egg output but relies on DNA from the larval or adult form of the parasite
in the human host, this technique may be a powerful tool for the early detection of infections,
especially during the prepatent phase. Both issues are currently under investigation and
preliminary data obtained from the murine model reveal promising results.
Financial support: Fiocruz, CNPq
67
IMMUnOdIagnOSIS Of SCHIStOSOMIaSIS
Peralta JM, gonçalves MML, Cavalcanti Mg.
Instituto de Microbiologia and Hospital Universitario Clementigo Fraga Filho, UFRJ and
Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil
Schistosomiasis remains one of the most prevalent parasitic infections in the world. Since
1984, the World Health Organization introduced a strategy for morbidity control which had
become feasible because of the availability of effective and safe single dose of schistosomicidal
drugs. The mainly strategy adopted by the Brazilian governmental control program of
schistosomiasis has been the screening of infected individuals by parasitological fecal exam
and treatment of positive cases. In this manner it was able to keep reasonably cost-effective
for the past four decades. However, in spite of the decrease of morbidity and endemicity
(decreased numbers of infected individuals) in several areas where mass treatment was
successful, the risk of infection remained practically untouched. It is well known that samples
from individuals asymptomatic or with mild infection or previously treated frequently result
in false negatives because the reduced sensitivity of the traditional diagnosis based on the
detection of Schistosoma eggs in stool or urine. New approaches to refine the diagnosis have
been considered, mainly because light infections might be undetected and as a consequence
it would jeopardize control of the parasite transmission. The diagnosis of Schistosoma
infection is based on coproscopic examinations, but immunological analysis using different
assays complement the diagnosis. Nonetheless, at least two main facts have limited the use
of immunological assays in the diagnosis of Schistosoma infection: cross reactivity with other
helminthiasis and persistence of antibody response after treatment. In fact, cross reactivity
was described in cases of ancylostomiasis and ascaridiasis in few studies that were not able
to rule out schistosomiasis. The persistence of elevated level of antibodies detected in posttreatment samples in the absence of egg-excretion is an obstacle to assure 100% cure. On
the other hand, around 15 to 20% individuals continue to eliminate parasite eggs despite
successive negative parasitological tests even after one or more cycles of treatment and in
some cases, adult worms might even remain in the mesenteric veins which could result in
low endemicity rates and maintained sample reactivity. Since Kato-Katz method has been
used as standard to evaluate other diagnostic methods (including immunoassays) to detect
Schistosoma infection, development of new tests is compromised because low sensitivity of
parasitological exams fail to detect low intensity infection which results in poor diagnosis and
questionable cure criteria. How to diagnose schistosomiasis in individuals with low intensity
infections? All together, the results point to the need of further investigation and higher
investments in test development for improving diagnosis and to define reliable markers of cure
after schistosomicidal treatment. Antibody-based assays generally have high sensitivities, but
serology is unable to discriminate between active and past S. mansoni infections. In the past
years, immunoassays like enzyme-linked immunossorbent assay (ELISA) and western blot
(WB) using crude or purified antigens derived from adult worms and eggs were developed
to determine anti-Schistosoma immunoglobulin (IgA, IgM, IgE , IgG and subclasses) levels.
Schistosoma – induced isotype response have a correlation with susceptibility/resistance to
infection and also to disease severity.. Studies show that increased IgG1 and IgE (effector
Ab) levels are associated to host resistance. In contrast, decreased levels of IgG4 and IgM
68
(blockers) indicate reduced susceptibility to infection. We standardized and evaluated the
ELISA and WB assays for the diagnosis of S. mansoni infection in N. squamipes naturally
living in Sumidouro (RJ) water streams. Furthermore, we determined the kinetics of antibody
production using serum samples obtained from rodents experimentally infected and after
specific treatment. As an additional diagnostic strategy, circulating antigens determination
such as CAA (circulating anodic antigen) and CCA (circulating cathodic antigen) by captureELISA is the most accurate immunodiagnostic test for active S. mansoni infection. However,
low sensitivity of the test is troublesome when stool samples present with low parasite load.
All above suggest that the use of immunological assays for seroepidemiological surveys
permits the evaluation of the prevalence and incidence rates of infection in endemic areas
and can also be used as a screening test in areas with low prevalence. Furthermore, ectopic
presentations such as genital and neuroschistosomiasis have been diagnosed more often in
migrants and non-resident individuals of endemic areas whose clinical manifestations are
associated to any parasite load even after years of the primary exposure. In most of this
case the coprological examination is negative and serological methods can be helpful for the
diagnosis.
69
UtILItY Of gEOSPatIaL tOOLS fOR COntROL Of PaRaSItIC dISEaSES
Robert Bergquist
Retired Medical Officer at the UNICEF, UNDP, World Bank, Special Programme for Research
and Training in Tropical Diseases (TDR), Ingerod, Brastad, Sweden
Geographical Information Systems (GIS) not only facilitates access to epidemiological
data through visualization in the form of maps, but can also be utilized for exploratory
analysis based on spatial statistics and the development of mathematical models of how
disease prevalence can change under the impact of various parameters. Moreover, data
emanating from earth-observing satellites regarding vegetation, land use, elevations, surface
temperature, rain fall, humidity, etc. complement GIS as the remotely sensed, continuously
updated information allows the analysis of disease distribution based on environmental
characteristics. However, already Hippocrates noted that certain diseases tend to occur in
some places and not in others so the idea that location influences health is far from new.
However, it was not until GIS, satellites and computer-assisted applications made it possible
to translate spatio-temporal data-sets of discovered phenomena into annotated maps that
the true capacity and versatility of this line of investigation came into its own. Its strength in
the field of epidemiology is based on the focus on the specific requirements of the causative
infectious agents, a fact which has proven particularly fruitful for the study of those parasitic
infections which depend on intermediate hosts for their transmission such as, for example
malaria and schistosomiasis. The geographical distribution of these diseases is limited by
the environmental requirements of their intermediate hosts/vectors, as well as the ambient
temperature in these hosts/vectors, two pieces of information which effectively govern
the spread and intensity of parasitic infections. Indeed, the concept of Growing Degree
Days (GDD), a parameter which now has been developed for many parasitic infections, is
the current base for the construction of reliable forecasts how the distribution of these
diseases can be expected to vary when the climate changes. This paper discusses the current
capability of satellite data collection in terms of resolution (spatial, temporal and spectral) of
the sensors onboard earth-observing satellites, drawing attention to the utility of computerbased models of the Earth for epidemiological research. Virtual globes, which are superior
to conventional maps, are available from Google and other commercial firms. The great
advantage with these contraptions is that they show geographical and man-made features,
but can be annotated with any type of other collected data, for example disease distribution,
demography, economy and other measures of particular interest for epidemiological study.
Disease surveillance and the early-warning schemes used in the veterinary field are examples
of practical, map-based systems now gaining importance in epidemiology.
70
vaCCInES tO COMBat nEgLECtEd tROPICaL dISEaSES: CURREnt StatUS Of tHE SMtSP-2 SCHISTOSOMA vaCCInE
Jeffrey Bethony
Department of Microbiology and Tropical Medicine, George Washington University Medical
Center, Washington, USA
The neglected tropical diseases (NTDs) represent a group of parasitic and related infectious
diseases such as amoebiasis, Chagas disease, cysticercosis, echinococcosis, hookworm,
leishmaniasis, and schistosomiasis. Together, these conditions are considered the most
common infections in low- and middle-income countries, where they produce a level of
global disability and human suffering equivalent to better known conditions such as HIV/AIDS
and malaria. Despite their global public health importance, progress on developing vaccines
for NTD pathogens has lagged, because of some key technical hurdles and the fact that these
infections occur almost exclusively in the world’s poorest people living below the World
Bank poverty line. In the absence of financial incentives for new products, the multinational
pharmaceutical companies have not embarked on substantive research and development
programs for the neglected tropical disease vaccines. Here, we review the current status
of scientific and technical progress in the development of vaccine against schistosomiasis.
We discuss animal testing (pre-clinical), product development, cGMP manufacture, and
proposed clinical development of the Sm-TSP-2 Schistosoma Vaccine.
71
HEaLtH EdUCatIOn and SCHIStOSOMIaSIS COntROL In MInaS gERaIS, BRazIL: nEW
SCEnaRIOS and CHaLLEngES
Cristiano Lara Massara, 2Martin Johannes Enk, 3virginia Schall
1
Laboratório de Helmintologia e Malacologia Médica, 2Laboratório de Esquistossomose,
Laboratório de Educação em Saúde e Ambiente - Centro de Pesquisas René Rachou/Fiocruz
Minas Gerais
1
3
Introduction: The increase of tourism, especially ecotourism, in Brazil since the year 2000
and internal migration are phenomenons which effect the geographic distribution of various
endemic diseases, among them schistosomiasis. In Minas Gerais ecotourism is attracting the
middle class into rural areas, prior destined to agricultural activities and nowadays shared
with small tourism ventures. These new scenarios increase the risk of infection for groups not
foreseen in the literature and by epidemiological determination. Fiocruz/Minas, as reference
institution for schistosomiasis in the state, was sought by groups of middle class people in
consequence of the occurrence of acute schistosomiasis. The difficulties of establishing the
diagnosis ‘schistosomiasis’ in health services used by these patients who often received
contradictive information about their illness, has been published. In the light of these
facts, the development of new strategies and materials which increase the divulgation,
not only about the disease, but also about its new economic and social determinants and
the vulnerability of populations with different profiles becomes eminent. This information
has to reach health professionals acting on the frontline of medical attendance, so that
the diagnostic hypothesis ‘schistosomiasis’, of which the acute form is often mistakenly
confused with other diseases, is considered. Also, a population, well informed about this
issue, could provide valuable hints for setting the diagnosis by relating the disease to tourist
environments that they frequented recently. Considering all above mentioned, a research
team of the CPqRR has been working in a multidisciplinary perspective, developing new
informative materials, offering courses for health professionals with an approach that
stimulates to work in an integrated manner, in which knowledge and praxis interact, with
the active participation of politicians responsible, community leaders and the general
population in order to promote sustainability of the interventions. In contrary to this, the
materials about schistosomiasis today available vacillate between a techno-scientific and
spectacular style with a preponderance of some sort grotesque esthetics and are merely
informative. They prioritize a form of pedagogic activity, which is similar to marketing
and publicity strategies – also characteristics of public health emergency campaigns - ,
reproducing copies of each other and repeating incorrect images and passed concepts. It
is of paramount importance to make material available written in simple language and with
images that can be used by health and education professionals for the comprehension of and
the approach to not only biomedical but also social aspects of schistosomiasis transmission
and maintenance. Materials of this nature have the potential to stimulate educative
meetings, during which popular and scientific knowledge can be shared, initiating and
promoting a permanent dialog which goes beyond transmission of information and leads to
a reflection about relations between health and socioeconomic and cultural development,
public policies and citizenship. These materials also serve to keep health professions aware
about the risk of schistosomiasis among until recently not affected population groups and to
72
keep the diagnosis schistosomiasis in mind order to identify cases of acute schistosomiasis.
Methodology: The development of educative materials and courses for permanent
education of health and education professionals is based on a shared creation process with
the target group about their knowledge and social representations about health and illness.
This process takes target group, objectives, topics and contents into account and considers
the dialog between health and educational perspectives as well as the field experience of
the authors. Simple language is chosen to transmit scientific knowledge with rich illustration,
contextualizing disease transmission in situations adapted to the local reality. The material
describes the track of the parasite in the human body (“Os caminhos da esquistossomose
dentro do nosso corpo”) and in the environment (Os caminhos da esquistossomose no meio
ambiente) informing about different stages and forms of the disease. This material can be
used for formal and informal education and by health professional during short courses
or routine work with the community. A third booklet denominated “X Tudo” offers games
and divertissements for application after working with the others. All three are available
at [email protected]. Results: The material printed by the State’s Secretary of Health is
being distributed to municipalities in endemic areas for utilization in projects, schools and
communities. Requests of other states have been registered, which requires a systematic
evaluation for its applicability. The material intends firstly to increase popular awareness by
stimulating community participation trough reflection about and comprehension of social
determinants involved in the process of schistosomiasis prevention and control and secondly
to actualize knowledge of health professionals related to the new epidemiological scenarios
of the disease, facilitating adequate diagnosis and treatment:
Financial support: PIDE (Programa Integrado de Esquistossomose) and CNPq
73
HEaLtH EdUCatIOn, COgnItIvE and COnCEPtUaL dEvELOPMEnt, and CHangE Of
attItUdE Of CHILdREn In a HELMIntHIC EndEMIC aREa In MInaS gERaIS, BRazIL
Maria flávia Carvalho gazzinelli1; aline da Silva Miranda2; Lucas Henrique Lobato de araújo3;
natércia acipreste Moura4; délcio fernando guimarães Pereira5; Jeffrey Michael Bethony6.
Professor Associado da Escola de Enfermagem da Universidade Federal de Minas Gerais
(UFMG); 2Mestranda da Escola de Enfermagem da Universidade Federal de Minas Gerais
(UFMG); 3Enfermeiro, apoio técnico do Projeto de Pesquisa; 4Professora do Curso de
Psicologia do Centro Universitário Newton Paiva em Belo Horizonte, MG; 5Professor do
Curso de Psicologia do Centro Universitário Newton Paiva em Belo Horizonte, MG; 6George
Washington University Medical Center, Washington, DC, USA.
1
Introduction: Epidemiological studies conducted in the State of Minas Gerais show high rates
of prevalence of infection by nematodes helminthes. Malnutrition and cognitive deficits
are considered the major pathology associated with helminth infections in schoolchildren.
This study aimed to evaluate the effects of two methods of health education on cognitive
development, conceptual development, and change of behavior and attitude of healthy
children and helminth-infected children who were subsequently treated for helminthes.
Methods: The study sample consisted of 98 children, 47 boys (48%) and 51 girls (52%),
6-10 years, in the region of Americaninhas, Novo Oriente de Minas Municipality. The
study involved three randomized groups of helminth-infected and healthy schoolchildren.
Group 1 (N = 33) received a non-directed educational approach based on the philosophy
of John Dewey, where theme such as helminthes, hygiene, health and environment were
discussed. Group 2 (N = 34) also received a directed educational approach, addressing
the same theme. The control group (N = 31) received no educational intervention. All
educational interventions occurred, between May and November 2009, with 9 interventions
per group. Questionnaires were applied before and after the final intervention, including
tests to evaluate the cognitive performance of the children. To assess their conceptual
development, a closed questionnaire covering general knowledge about health and specific
knowledge about helminthes and attitudes was applied. The open-questionnaire was used
to determine if there was a change of attitude of the children towards the infection and
infection-related behaviors after the intervention. Results: The intervention resulted in small
improvements in psychological performance tests, (without statistical significance), especially
in regards to changes in the classification level of cognition. While we observed significant
advances in all three groups., in regards to general knowledge, the greatest improvements
were seen for Group 2 (Transmission) followed by Group 1 (Dewey), followed by the Control
Group. discussion. The transmission pedagogical method (Group 2), while grounded in a
epistemological system that is questionable today, allows important learning in contrast with
the not directive pedagogy (Group 1). These results imply that a dialogue between the two
methods is needed to formulate a health education program that has an impact. Further
implications of these findings will be discussed during the presentation.
74
SCHIStOSOMIaSIS-RELatEd PERCEPtIOnS, attItUdES and tREatMEnt SEEKIng
PRaCtICES In MagU dIStRICt, tanzanIa: PUBLIC HEaLtH IMPLICatIOnS
J.R.Mwanga, MSc Medical Sociology, Research Scientist *, P. Magnussen, Md, Specialist in
tropical Medicine & Infectious diseases **, C.L. Mugashe, MSc Medical Parasitology, Research
Scientist * =, R.M. gabone, Md, MMed Pathology, Senior Research Scientist * =, J. aagaardHansen, Social anthropologist, Md, Specialist in general Medicine, dtM, dPH **
* National Institute for Medical Research, Mwanza Centre, P. O. Box 1462, Mwanza, Tanzania
** DBL-Centre for Health Research and Development, Thorvaldsensvej 57, 1871 Frederiksberg
C, Denmark
= Deceased
Introduction: Disease specific health education programmes should be based on knowledge
about perceptions, attitudes and treatment seeking behaviour of the affected populations.
A study on perceptions, attitudes and treatment seeking practices related to schistosomiasis
was conducted among the Wasukuma in the rural Magu district of Tanzania at the shore
of Lake Victoria where Schistosoma haematobium and mansoni infections are endemic.
Methodology: The study applied in-depth interviews, focus group discussions and a
questionnaire survey among adults and primary school children. Results: The perceived
symptoms and causes were incongruous with the biomedical perspective and a number
of respondents found schistosomiasis to be a shameful disease. Lack of diagnostic and
curative services at the government health care facilities was common, but there was
a willingness from the biomedical health care services to collaborate with the traditional
healers. Conclusions: Recommendations to the District Health Management Team were:
that collaboration between biomedical and traditional health care providers should be
strengthened and that the government facilities’ diagnostic and curative capacity with regard
to schistosomiasis is upgraded. Culturally compatible health education programmes should
be developed in collaboration with the local community.
Financial support: DBL-Centre for Health Research and Development.
75
SCHOOL-BaSEd aCtIOnS fOR SCaLIng-UP COntROL Of SCHIStOSOMIaSIS In tHE
EndEMIC aREa Of PERnaMBUCO, nORtHEaStERn BRazIL, WItHIn tHE UnIfIEd
HEaLtH SYStEM
tereza C. favre; ana Paula B. Pereira, Lilian CnH Beck, aline f. galvão, Otávio S. Pieri
Laboratório de Ecoepidemiologia e Controle da Esquistossomose e Geohelmintoses, Instituto
Oswaldo Cruz, Fiocruz, Rio de Janeiro.
Introduction: Resolution CD49.R19 of the Pan-American Health Organization (PAHO)
of October 2009 urged Member States to eliminate or reduce schistosomiasis and other
neglected diseases related to poverty for which tools exist, to levels so that these diseases
are no longer considered public health problems by 2015. Accordingly, the new guidelines
of the Ministry of Health (MS) for endemic areas of schistosomiasis with prevalence above
5% recommend biennial stool surveys of schoolchildren in urban localities and of whole
populations in rural localities giving priority to school-aged children (6-15 years). The
MS also recommends prompt treatment of the positives as well as health education and
environmental sanitation. According to the Computerized System of the Schistosomiasis
Control Programme (SISPCE), from 2001 to 2008 the Basic Attention / Family Health (AB/SF)
teams carried out 828,365 active-search examinations in the endemic area of Pernambuco
state, covering only 10.3% of the total population at risk (8,017,479 according to the Locations
Information System – SISLOC) in that area. Of the 99 municipalities surveyed at least once
during that 8-year period, 58 (58.6%) had prevalence above 5% and are thus entitled to
biennial stool surveys. From a total of 5.7 million residents, 4.7 million are estimated to
live in urban localities and 1.0 million in rural localities. As the school-aged population
represents 26.2% of the total population in that area, the AB/SF teams of the municipalities
involved should carry out at least 1.5 million examinations biennially to accomplish the MS
recommendations. The purpose of this work is to evaluate the effectiveness of the school
setting in delivering diagnosis and treatment to school-aged children, as compared with the
community-based delivery, aiming to scale-up schistosomiasis control in that area. Methods:
The municipality selected for study was Araçoiaba, occupying an area of 96 km2 in the
Metropolitan Region of Recife, and located 49 km North of the capital. All children aged
6-15yrs who were enrolled in the 10 public schools of Araçoiaba in 2009 were considered as
the catchment population, totalling 3,273 (95.8% of the total population in this age-group).
All public schools of the municipality were ranked along with the number of enrolled children
and paired by area (rural or urban). Two sets of five matched schools were formed. Each
set was randomly assigned either to the school-based or the community-based delivery
scheme. The research team monitored the activities of the AB/SF teams to assure that
the standard procedures recommended by the MS were followed in either setting. Those
procedures included: (i) distribution of stool vials and collection in the following days, (ii)
identification of the infected children through fecal examination by the Kato-Katz method,
(iii) forwarding the positives for medical evaluation and drug prescription, (iv) administration
of a single oral dose of praziquantel (PZQ) 60 mg/kg under medical supervision. The patients
identified through the school-based setting were scheduled for treatment at their schools,
whereas those from the community-based setting were scheduled for treatment at the
nearest Basic Health Unit (UBS). Two follow-up surveys were conducted: at four and 12
76
months after treatment. Results: At baseline, a total of 646 (26.6%) out of 2,467 children
were positive for Schistosoma mansoni. Coverage rates of the community-based scheme
were 98.9% for stool vial distribution and of 84.9% for diagnosis, which are significantly
higher (p <0.05) than those of the school-based scheme (96.4% and 74.8%, respectively).
However, the school-based scheme had significantly higher ratios of daily distribution (62.4
children per day - cpd) and collection (46.7 cpd) of stool vials than the community-based
scheme (27.3 cpd and 23.2 cpd, respectively). There was no significant difference (p>0.05) in
treatment compliance between the two settings (88.7% in the community and 90.4% in the
school), but the average time spent by the AB/SF teams with treatment in the school was
significantly lower than in the community. At four months after treatment 51 (9.1%) out of
563 children examined were egg-positive. There was no significant difference in diagnostic
coverage between the community (93.3%) and the school (92%) schemes. At one year after
treatment a total of 139 (26.4%) out of 527 children examined were positive. The diagnostic
coverage in the community (92%) was significantly greater than in the school (78%), but
there was no significant difference in treatment compliance between the school (86.3%) and
the community (79.5%) settings. Conclusion: An approach is thus proposed to enable shortterm improved access to and coverage of the control actions targeted at this particularly
vulnerable high-risk group, combining school-based and community-based interventions as
well as preventive measures to reduce transmission.
Financial support: NTC/PCT/WHO
77
HEaLtH SERvICES aCCESSIBILItY, UtILIzatIOn and KnOWLEdgE and PERCEPtIOn
Of SERvICE PROvIdERS and USERS RELatEd tO SCHIStOSOMIaSIS COntROL In an
EndEMIC RURaL aREa In MInaS gERaIS, BRazIL
andrea gazzinelli
Escola de Enfermagem da UFMG, Belo Horizonte, MG
Introduction: Little is known about responses to schistosomiasis by local providers and users
of health services. This paper examines patterns and factors in accessibility and utilization
of schistosomiasis diagnostic and treatment services, as well as knowledge and perceptions
of residents, health workers and health officials of schistosomiasis control in São Pedro
District in Jequitinhonha Municipality in Minas Gerais State, Brazil. São Pedro District is a
poor area where more than half of the population receives government assistance (Bolsa
Família). Methodology: A questionnaire survey was carried out in 2007 with 1,228 members
of 334 households (more than 95% of all households in the District) about health services
accessibility and utilization in relation to socioeconomic status, social networks, signs and
symptoms, as well as health behavior retrospectively for the period 2002-2006. Focus group
discussions were held among 30 adult males and females and interviews done with the five
staff members of São Pedro Health Center and two municipal health officials on knowledge
and perceptions regarding schistosomiasis and its control. Stool examinations were done
for all study members using the Kato/Katz method. The epidemiological data were analyzed
using univariate and multivariate methods and the perception/knowledge information
using the content analysis method by Bardin (1991). Results: The ratio for utilization of
diagnostic services between 2002-2006 were significantly lower for males than females, as
well as persons below 34 years of age and per capita income of less than 60 Reais a month
and not owning a car/motorcycle, including households receiving government assistance,
households having more than 0.80 persons per room and persons with or without symptoms
but infected with S. mansoni. All these variables except age remained significantly associated
in the multivariate model. Only 24.5% persons self-reporting to a health facility between
2002 and 2006 obtained a stool test from the local health center or other health facilities.
One hundred thirty eight of the 197 (70.0%) persons experiencing symptoms suggestive of
schistosomiasis 30 days prior to the 2007 survey used home remedies. The analysis of patient
records for the 12 month period prior to this survey showed that only 4 of the 179 patients
with symptoms suggestive of schistosomiasis received praziquantel, the others being treated
symptomatically for soil-transmitted helminths. Only 18.3% of the persons who obtained
a stool test and half of those who were treated obtained these services through the local
health center. The others used health facilities outside the study area. Residents were aware
of the relationship between the deficiency of the water supply system, schistosomiasis and
low motivation to seek chemotherapy because of side effects of praziquantel, reinfection
and accessibility problems due to high travel costs and lack of a full-time physician and a
laboratory at São Pedro Health Center. The health providers, by contrast, attributed the
problem of schistosomiasis in the study area largely to the low educational level of the
population and health services deficiencies. Conclusions: This study confirms recent reports
of low accessibility and utilization of schistosomiasis diagnostic and treatment services in
other endemic areas in Brazil and recommends that 1) health centers include laboratory
78
facilities and strengthen health education programs with active community participation
and 2) that more research be carried out on the knowledge and perception of providers
and users of health services to better evaluate needs and constraints surrounding healthseeking behaviour and the accessibility of health services in poor rural areas. Financial
support: UNICEF/UNDP/World Bank/WHO/TDR, Fogarty International Center Training Grant
(1D43TW006580), FAPEMIG, INCT-DT, TDR/WHO, CNPq.
79
CHROMatIn REgULatIOn In SCHIStOSOMES and HIStOnE-MOdIfYIng EnzYMES aS
dRUg taRgEtS.
Raymond J. Pierce; Stéphanie Caby; florence dubois; Julien Lancelot; Jacques trolet; Céline
Cosseau; Christoph grunau; guillaume Mitta; Luiza f.a. almeida; Leila nahum; guilherme
Correa de Oliveira;
CIIL, Inserm U1019, CNRS UMR 8204, Institute Pasteur de Lille
Introduction: Histone modifying enzymes (HME) are central actors in the regulation of the
epigenetic modification of chromatin and aberrant epigenetic states often associated with
cancer led to interest in HMEs as targets for therapy. Among these the histone deacetylases
(HDAC) have been the subject of intense study and a number of HDAC inhibitors (HDACi) are in
clinical trials. HDACs deacetylate acetylated lysine residues in a variety of proteins, including
histones, but also transcription factors and cofactors, as well as non-nuclear proteins such
as tubulin. Of the three main classes of HDACs in eukaryotes, enzymes of classes I and II
share similar catalytic domains and a Zn2+-dependent mechanism, whereas the class III
enzymes, the sirtuins, are phylogenetically unrelated and have an NAD+-dependent catalytic
mechanism. Broadly, HDACi induce cell death in cancer cells via apoptosis but they can also
act on the cell cycle, on tumour angiogenesis or via the regulation of host cell responses.
HDACi have also stimulated interest as anti-parasitic drugs and have been tested against P.
falciparum, Toxoplasma gondii and the major kinetoplastid parasites. We have characterized
and studied the HDACs of Schistosoma mansoni and have shown that HDACi cause the
death of S. mansoni larvae and adult worms in vitro via chromatin hyperacetylation and
the induction of apoptosis. Methodology: S. mansoni HDACs were identified by mining the
genome sequence and full-length coding sequences were validated using RACE-PCR. Their
identity was verified by phylogenetic analysis and their expression during the parasite lifecycle was determined using quantitative RT-PCR. The functional activity of selected HDACs
was tested using reporter gene assays in mammalian cell lines. The effect of HDACi was
determined by measuring their effect on the viability of schistosomula and adult worms and
the induction of apoptosis assayed by the TUNEL method and the induction of caspase 3/7
activity. The overexpression of HDAC target genes after HDACi treatment was determined
using qRT-PCR and correlated to hyperacetylation of the corresponding proximal promoters
using quantitative chromatin immunoprécipitation (qChIP). Native acetylation of H3K9
on the promoters was determined using ChIPSeq. Results: S. mansoni possesses 3 class I
(HDAC1, 3 and 8), 4 class II (HDAC4, 5, 6 and 10) and 5 class III (Sirt1, 2, 5, 6 and 7) HDACencoding genes. Several of the corresponding coding sequences, including SmHDAC8 and
SmSirt1 possess insertions in their catalytic domains compared to mammalian orthologues.
Functional assays show that SmHDAC1 represses gene transcription in reporter gene assays
and that SmSirt1 potentiates the activity of the transcription factor FoxO, indicating the
conservation of HDAC functions in S. mansoni. Treatment of schistosomula or adult worms
with HDACi such as TSA or SAHA (inhibitors of classes I and II) or sirtinol (a sirtuin inhibitor)
induces the death of both larval (schistosomula) and adult worms and this is preceded in the
larvae by the induction of apoptosis as measured by TUNEL staining and the increase in the
activity of caspase 3/7. Moreover, such treatments induce a rapid increase in the general
level of histone acetylation, particularly of H4. This in turn correlates with the overexpression
80
of certain genes, including those encoding caspases 3 and 7. ChIPSeq analysis showed that
H3K9 acetylation of the caspase 7 promoter in untreated parasites was very low. Finally,
qChIP analysis shows that the proximal promoter of caspase 3, and in particular of the
caspase 7 gene show hyperacetylation of histone H4 after HDACi treatment. Conclusions:
These results lead us to consider that schistosome HDACs, as well as other HMEs, are
promising targets for the development of new drugs against schistosomiasis. To this end,
a project (SEtTReND) supported by funding from the EC has been initiated, with the aim of
characterizing the HMEs of S. mansoni, particularly those involved in histone acetylation/
deacetylation and methylation/demethylation. These enzymes will be validated as targets
and specific inhibitors will be identified against selected enzymes that could be candidates
as lead compounds for drug development.
Financial support: EC (FP7-Health); Inserm-Fiocruz; Grant ANR-07-BLAN-0119-02
81
MaLE-fEMaLE IntERaCtIOn: a tRanSCRIPtOMIC vIEW
giulliana t almeidaa, felipe C. f. Beckedorffa, Murilo Senaa, Ricardo deMarcob and Sergio
verjovski-almeidaa
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05508-900
São Paulo, SP, Brazil, bDepartamento de Física e Informática, Instituto de Física de São Carlos,
Universidade de São Paulo, 13560-970 São Carlos, SP, Brazil.
a
Introduction: Schistosoma mansoni is a dioecious parasite and adult worms display a high
level of sexual specialization. Because Schistosomes are one of the simplest dioecious
organisms, the study of their sexual differentiation may provide clues for understanding
the evolution of this process. A vaccine that inhibits both development and fertility of
schistosomes would be useful since it should reduce both morbidity and mortality in
the human population; it would also stall the transmission of the disease, as fecal egg
count would decrease significantly. Therefore, further understanding of the molecular
mechanism of sexual differentiation and development of sexual traits may provide ways
to control the parasite by limiting its reproduction rate. Methodology: We investigated
the influence of pairing in the expression profiles of S. mansoni male and female adult
worms. To this end, we performed an in vitro culture of paired worms and separated
male and female adult worms. After 13 days of cultivation mRNA was extracted and
amplified. We utilized a 44.000 elements microarray to compare expression profiles
of paired and unpaired worms from each sex. Additional microarray experiments with
mRNA extracted from re-paired worms and worms maintained in the same culture
medium, but separated by a permeable support (8µm), were performed to evaluate
the reversibility of gene expression alterations after separation of couples. For further
exploration of the male and female adult worm transcriptome, we also constructed and
sequenced libraries from mRNA of female and male adult worms by pirosequencing
with the 454 Genome Sequencer FLX System. Results: We detected 217 and 2043 genes
with differential expression between paired and unpaired worms in male and female
adult worms, respectively. In addition, re-pairing during 7 days of separated worms for
six days caused 494 genes with differential expression in separated female worms to
return to the levels observed in paired worms. The same change in expression levels was
not observed in separated worms that were put back together with a permeable wall
between each pair, which still display an expression profile similar to that observed in
separated worms. 454 sequencing reaction of a male library resulted in ~280k sequences
with average length of 99 bp and two sequencing reactions using the female library
resulted in ~283k sequences with average length of 106 bp. Analysis of these sequences
revealed that the male and female pirosequencing sampled 7149 and 4657 of the 12.850
genomic models described, respectively. 3992 genomic models have been sampled in
both male and female sequencing, while 3157 and 655 genomic models have only been
sampled in the male and female sequencing, respectively, and may represent genes
displaying sex-specific transcription. Conclusions: Our data shows that separation of
mature adult couples leads to a considerable change of the expression profiles of these
worms. This suggests that sexual stimulation plays an important role in the genetic
program of Schistosoma. Changes in expression profiles caused by separation of worms
82
were partially reverted after repairing and seven days of in vitro cultivation. The same
changes were not observed in worms placed in the same culture medium, but unable to
establish physical contact. These data suggest that stimulation for differential expression
of paired worms mostly from physical stimulus rather than a diffusible factor. In addition,
pirosequencing permitted us to propose several genes with sex-specific transcription,
which may play an important role in the interaction between male and female adult
worms.
83
PROgRESS WItH SCHIStOSOME tRanSgEnESIS
Paul J. Brindley, gabriel Rinaldi, Sutas Suttiprapa, victoria H. Mann
Department of Microbiology, Immunology and Tropical Medicine, School of Medicine &
Health Sciences, The George Washington University, 2300 Eye Street NW, Washington, DC
20037 USA; [email protected]
Introduction: Draft genome sequences for Schistosoma japonicum and S. mansoni are now
available. The schistosome genome encodes ~13,000 protein encoding genes for which the
function of only a small minority is understood. Many of the new genes will represent novel
intervention targets. We consider there is a valuable role for transgenesis in functional
genomics investigation of these new schistosome gene sequences. In gain-of-function
approaches, transgenesis can lead to integration of reporter transgenes into the schistosome
genome which, among other uses, can facilitate insertional mutagenesis screens. By
contrast, transgene driven, vector-based RNA interference offers potentially powerful lossof-function manipulations. Methodology: Recent research in our laboratory has focused on
development of tools and methods to facilitate schistosome transgenesis. We investigated
the utility of retroviruses and transposons to transduce cultured schistosomes. Results: We
found that vesicular stomatitis glycoprotein (VSVG) pseudotyped murine leukemia virus
(MLV) can transduce developmental stages of S. mansoni including eggs. We have also
observed that the piggyBac transposon is transpositionally active in schistosome tissues.
Approaches with both VSVG-MLV and piggyBac have resulted in somatic transgenesis,
and in particular have lead to integration of active reporter transgenes into schistosome
chromosomes. Conclusion: Our approaches have lead to the first reports of integration of
reporter transgenes into schistosome chromosomes or indeed into the chromosomes of
any parasitic worm. Our experience with these systems will be reviewed, along with recent
findings with retroviral transgene mediated RNA interference and germ line transgenesis.
Financial support: NIH-NIAID award RO1 AI072773
84
SCHIStOdB.nEt: SCHISTOSOMA MAnSOnI gEnOMIC and fUnCtIOnaL gEnOMICS
data IntEgRatIOn
guilherme Oliveiraa,b*, Mariana C. Simõesb, fabiano Sviatopolk Mirsky Paisb, Roney Coimbrab,
adhemar zerlotinib
René Rachou Research Center, Oswaldo Cruz Foundation – FIOCRUZ, Belo Horizonte,
Brazil; bCenter for Excellence in Bioinformatics, Oswaldo Cruz Foundation – FIOCRUZ, Belo
Horizonte, Brazil.
a
Introduction: Advances in structural and functional genomics, proteomics, genetics and
molecular biology have substantially increased the amount of available data for schistosome
research. The integration of the information to make it usable for the community in an easy
to navigate and searchable format is an absolute necessity. SchistoDB 2.0 currently includes
the genomic data, approximately 13.000 gene models and annotation and integrates EST
and metabolic mapping. SchistoDB offers a variety of tools including BLAST, protein motif
searches, keyword searches of pre-computed BLAST results, Gene Ontology assignments,
protein family information and microarray probes. Interaction with the database occurs via
a single user-friendly web interface. Here we describe the production of SchistoDB version
3.0. Methodology: For integrating additional data we used the available relational database
schema GUS (Genomics Unified Schema) already implemented. Microarray, SAGE and ChipSeq data was obtained from the literature and our own work. Interaction with the authors
was important to appropriately format the information. A database called SchistoCyc was
created using SRI PathwayTools software. The results are displayed with additional search
capabilities included in a similar web interface. Results: SchistoDB (www.SchistoDB.net)
is a genomic database for S. mansoni recently uploaded data significantly incrases to the
resources available. The combination of an annotated genome and a relational architecture
has facilitated the integration of the genome with other types of data and permitted the
construction of automated analysis pipelines. Also integrated into SchistoDB are SchistoCyc
and KEGG DRUG. Within SchistoCyc 112 metabolic pathways were predicted based on
the genomic data and is useful for the study of parasite biology and select reactions that
might play a crucial role in parasite metabolism. KEGG DRUG contains chemical structure
and data on drugs that target orthologous proteins of other organisms. We have also
linked gene products to the TDR Targets database. SchistoDB allows the user to save and
combine queries using Boolean logic. Conclusion: SchistoDB has been heavily used by
the Schistosoma research community and is reliable and powerful resource. Currently it
is extremely difficult, if not impossible for most groups, to access published information.
SchistoD offers easy access to the data. We plan to continue to add further functional
genomics data including a large number of experiments using next generation sequencing
and also genetics information. For version 3.0 a new web interface will be implemented. We
are also in contact with the research community to stimulate the use of the resource for prepublication purposes to permit data mining through SchistoDB.
Financial support: NIH (TW007012), SECTES/FAPEMIG (1181/08), CNPq (306879/2009-3 and
573839/2008-5).
85
POPULatIOn StRUCtURE Of S. MAnSOnI PRE- and POSt-tREatMEnt
Ronald E Blanton, Walter a Blank, Jackson M Costa, theomira M Carmo, Eliana a Reis, Luciano
K Silva, Lúcio M Barbosa, Mitermayer g Reis
Case Western Reserve University, USA
Centro de Pesquisas Gonçalo Moniz, Fiocruz, Salvador, Bahia
Introduction: We sometimes think of Schistosoma mansoni in endemic areas as a single
organism, whereas the parasite naturally exists as heterogenous populations. There is
evidence that these populations are structured, i.e. gene flow is limited geographically
and demographically. The control strategy recommended by the WHO is repeated
treatment with praziquantel since the prevalence quickly returns to pre-treatment levels.
The effect of widespread treatment on schistosome populations has been little studied,
although understanding who gets infected with which parasites, how populations
recover and why could be useful for planning public health strategies. Praziquantel has
been used since 1979, and currently is the only drug in production for schistosomiasis.
Under laboratory conditions, resistance to praziquantel appears relatively easy to
induce. In practice, 10-15% of those treated will continue to excrete eggs. These are
not considered treatment failures, but it is not understood if this represents selection
of a relatively resistant population or incomplete elimination without selection, such as
the presence of immature worms from recent infection. We used population genetic
analyses to understand gene flow in populations of schistosome infecting humans
and to test whether persistent parasites were drawn from the same population as
susceptible parasites. Methodology: Individuals from two small Brazilian communities
(pop. 481 and 367) separated by 12 km were interviewed for demographics, and 96%
(815) provided stools for examination, 92% (787) provided 3 stools. All subjects were
treated, reexamined 4-6 weeks later. Eggs were isolated from the whole stool pretreatment and post-treatment for those still egg positive (persistent infection). DNA was
extracted from the aggregate of all eggs collected, quantified by qPCR and genotyped at
15 microsatellite markers. Jost’s D was calculated to measure differentiation between
populations (genetic differentiation - <0.05 negligible, 0.051 – 0.24 moderate, >0.25
very great). Results: One third of the inhabitants had been previously treated (67%
praziquantel, 33% oxamniquine). The average prevalence of S. mansoni infections
was 41% with a typical age intensity profile. All 3 exams were egg positive for 124
individuals, and DNA could be obtained from 122 of these samples. There were 36
persistent infections (14%) The mean differentiation between samples from different
days was 0.019, compared to 0.012 for differentiation for replicates of the same
sample. The person-to-person differentiation was moderate at 0.085 and 0.126 for the
2 communities, which was higher than the differentiation between the 2 communities
as a whole (D = 0.047). Cured and persistent parasites appeared to belong to the same
population for each community (D = 0.007, 0.009). Conclusions: We found that: 1) there
is a consistent day-to-day excretion of genotypes in eggs over a 7 day period, 2) there
is substantial differentiation of parasite populations within individual hosts, 3) there is
geographic structuring of parasite populations over short distances in this rural area, 4)
simple persistence after treatment is probably not due to selection, since parasites in
86
the persistent population are not differentiated from the susceptible population. This
approach has been found sensitive enough to show small but consistent differentiation
between different lots of parasites even from the same laboratory life cycle. We will
continue to follow this population over 2 more treatment cycles for evidence of selection
and to understand how schistosome populations recover.
87
RESIStanCE Of BIOMPHAlARIA tO SCHISTOSOMA MAnSOnI InfECtIOn
Raquel Lopes Martins Souza
Universidade Federal de Alfenas-MG, Brasil
Biomphalaria is fresh water Planorbid of great medical relevance as intermediate host of
Schistosoma mansoni, a trematode parasite that causes human schistosomiasis, which
disease affects about 8 million people in Brazil. Successful parasitism of snail Biomphalaria
species and different strains by S. mansoni, could depend upon the penetration of miracidia
larvae with compatible host, establishment and production of cercariae. In the interaction
between S. mansoni and Biomphalaria, resistance or susceptibility of the snails to parasite is
determined by the genetics of both. Regarding moluscs, they have an internal defence system
(IDS) able to protect themselves against infections by potential pathogens. Recent studies have
demonstrated many similarities between the innate defense response of vertebrates and the
internal defense system of invertebrates, being identified in various invertebrates organisms
production of complement-like proteins, anti-microbial peptides, pattern-recognition
receptors (PRRs) such as toll-like receptor and C-type lectins, phagocytic cells, production of
highly toxic metabolites of oxygen and nitrogen. Biomphalaria snails responses to immune
challenge are lymphoid-independent, mediated by hemocytes (phagocytic cell population) in
cooperation with humoral components. In snails, circulating hemocytes are the principal line of
cellular defence involved in destruction of S. mansoni larvae inside the intermediate host, thus
determining the resistance to parasite. Direct evidence of the haemocyte participation on the
S. mansoni infection control was provided by histopathological analysis and experiments that
transferred the Amoebocyte-producing organ (APO) from resistant to susceptible snail strains.
The characterization of circulating haemocytes in Biomphalaria has been described by various
authors, focusing on morphological, functional and biochemical aspects of these cells. Studies,
associated with hemocytes, have been indicated that soluble elements of the haemolymph
participate in the protective mechanism. Susceptible strain of B. glabrata previously inoculated
with cell-free haemolymph from resistant snails had lower infective rate after exposure to S.
mansoni miracidia. It was reported the presence of substances in B. glabrata hemolymph that
promote agglutination of hemocytes surrounding S. mansoni sporocysts, suggesting that soluble
factors of hemolymph participate in the recognition mechanism and opsonization of particles
by hemocytes. Experimental evidences suggested that Lectins/carbohydrate binding can be
involved in the S. mansoni recognition and activation of Biomphalaria hemocytes, and thus it
would be associated with snail resistance against the parasite infection. So the B. tenagophila
Taim (coming from the Ecological Station of Taim, state of Rio Grande do Sul, Brazil) studied by
our research group, represents an important experimental model, showing absolute resistance
to infection by S. mansoni. The results obtained in our laboratory allow the establishment of
the hypothesis that the resistance against S. mansoni is due to a specific characteristic of their
innate mechanism of defence and not to a mere physiological incompatibility between parasite
and host. The comparison between haemocyte responses to S. mansoni infection in resistant
and susceptible snail strains suggested that resistance observed in Taim strain is associated
with intense haemocyte activation, migration to the infection site, and the presence of specific
lectins and other factors in its hemolymph.
88
InfECtIvItY EvaLUatIOn Of BIOMPHAlARIA COuSInI USIng tHREE dIffEREnt
StRaInS Of SCHISTOSOMA MAnSOnI
Roberta Lima Caldeiraa, tatiana Maria teodoroa, Liana Konovaloff Jannotti-Passosb & Omar
dos Santos Carvalhoa
Laboratório de Helmintologia e Malacologia Médica e bMoluscário Lobato Paraense Centro
de Pesquisas René Rachou, Fiocruz, Belo Horizonte, Brasil
a
Introduction: Previous investigations using phylogeny, morphological and molecular
taxonomy, of Brazilian, Bolivian and Colombian snails identified as Biomphalaria amazonica,
showed that some specimens belonged to the species Biomphalaria cousini. These species
are undistinguishable through morphological characterization. This study described for the
first time, the presence of B. cousini in Brazil. On one hand no studies have been conducted
on the status of susceptibility of this species. On the other hand B. amazonica from Careiro
da Várzea (AM) showed to be susceptible to infection with two strains of Schistosoma
mansoni (BH and SJ) with an infection rate of 48% and 73% respectively. In this context, the
aim of the present work was to verify the susceptibility of B. cousini to different strains S.
mansoni. Methodology: A total of 90 specimens of F1 generation of B. cousini from Benjamin
Constant, state of Amazonas, kept in the René Rachou Research Center were exposed to S.
mansoni miracidia of the LE strain. The snails were individually exposed to eight miracidia.
As control were used 50 specimens of B. glabrata from Belo Horizonte, state of Minas Gerais,
individually exposed to eight miracidia of the S. mansoni LE strain and 10 specimens of B.
glabrata and 10 of B. cousini without infection. On the 30th day after exposure, and then
every week, the snails were singly placed in vials with water and exposed to the artificial
light (28 °C) to induce shedding of cercariae. If any specimen died during the experiment
it was examined by LS-PCR to detect the presence of S. mansoni DNA. The specimens that
survived 80 days after exposure without shedding cercariae were dissected and examined
by microscopic for cercariae and/or sporocysts. The specimens that shed cercariae were
exposed to light for 2 hours on alternative days until they died, and the cercariae were
inoculated in Swiss albino mice to verify the viability of them. This procedure was repeated
for SJ and AL S. mansoni strains. Results: B. cousini proved to be susceptible to the LE, SJ
and AL strains of S. mansoni with an infection rate of 31.6%, 5.6% and 3.4% respectively.
Conclusions: B. cousini is a potential host of the S. mansoni since this species showed to be
susceptible to all strains of the trematoda used in the experiment. Therefore, the presence of
this species in areas free of schistosomiasis represents a risk of introducing the disease under
circumstances that are favorable for its transmission. This issue gains even more importance
considering that B. cousini has been found in water collections located in the Amazon region,
which has been attracting workers, including some from schistosomiasis endemic areas.
Supported by PAPES V CNPq/Fiocruz
89
SUSCEtIBILItY Of BIOMPHAlARIA SPP fROM Man-MadE LaKES tO InfECtIOn BY
dIffEREnt StRaInS Of SCHISTOSOMA MAnSOnI
Silvana Carvalho thiengo
Laboratório de Malacologia, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro
In Brazil the studies on the occurrence of different strains of Schistosoma mansoni started in
the sixties when it was observed that miracidia from Paraiba river valley, state of São Paulo,
easily infected Biomphalaria tenagophila from the same region, but were almost unable to
infect the most susceptible neotropical species, Biomphalaria glabrata. Further studies not
only revealed morphological, physiological and genetic differences between both strains but
also pointed out the occurrence of other strains adapted to local intermediate snails. During
the last forty years 8 different S. mansoni strains have been maintained in the Laboratorio
de Malacologia do Instituto Oswaldo Cruz what made possible studies on the compatibility
between the parasite and species of Biomphalaria. Most of those studies used descendand
snails of those specimens collected in areas impacted by water resources development
projects as hydroelectric power stations. We report here the results of experiments on the
susceptibility of Biomphalaria straminea, B. occidentalis and B. amazonica from four dams
in central Brazil: Serra da Mesa, Peixe Angical, São Salvador and Manso to different strains
of S. mansoni. Methodology: In all experiments the specimens were individually exposed to
5 Schistosoma mansoni miracidia and the aquaria were kept at a room temperature of 2426oC throughout the experiment. Snails were observed daily and, if any specimen happened
to die, it was fixed in Railliet-Henry’s fluid and examined for developing stages of the
schistosome. This procedure provided the mortality index and to characterize the duration
of precercarial period all specimens were exposed to artificial light at five-days intervals after
25th day to exposition. The snails that survived for 60 days after exposure without shedding
cercariae, were fixed, dissected and examined to characterize the infection index in the each
colony. As for B. straminea 2,449 specimens (2-7mm in shell diameter) were exposed, from
54 collection stations in the area of influence of three hydroelectric dams (UHE Serra da
Mesa, UHE São Salvador and UHE Peixe Angical). For B. occidentalis and B. amazonica they
were used 222 and 257 specimens, respectively, with 2-5mm in shell diameter (for the first
species) and 3-8mm for the second one. As for the schistosome strains they were used:
three strains (CM, CMO and EC) isolated from areas of Northeast Brazil where B. straminea
has been reported as a vector; the PB strain from feces obtained in Padre Bernardo, Goiás
state, an urban focus of schistosomiasis approximately 300km from the lake of the UHE Serra
da Mesa; and BH and SJ strains of the Minas Gerais and São Paulo states. Results: Serra da
Mesa and S. Salvador: among 1,135 specimens used 15 became infected (infection index
of 1.3%) and 8 populations were susceptible to the schistosome strains: B. straminea from
Campinorte (Castelão, susceptible to CM and EC strains, and Planeta Água, EC strain), Colinas
(Tocantinzinho river, CM and EC strains). Minaçu (Canabrava river, EC strain), Niquelândia
(Codemin, CM and PB strains, and Almas river, CM strain), Uruaçu (touristic area, PB strain)
and Santa Rita do Novo Destino (Maranhão river, CM and EC strains). Manso: Of 257 B.
amazonica used, 17 became infected (infection index of 6.61%) and all specimens of B.
occidentalis proved unsusceptible. According to the strains used, of the 158 snails exposed
to BH miracidia, 6 became infected (3.79%); of the 44 exposed to SJ miracidia, 6 became
90
infected (13.63%); and of the 55 B. amazonica exposed to EC miracidia, 5 became infected
(9.09%). Peixe Angical: Among the 1,314 specimens used, eight had been infected (infection
index of 0.6%) with only the BH strain, (ii) for B. straminea populations, the mortality index
was 6.8% and, depending on the strain used, the indexes were 4.6%, 8.49% and 19% with
BH, CM and CMO strains, respectively, (iii) the infection indexes varied according to the B.
straminea populations, ranging from 0-12.5% and (iv) the duration of the precercarial period
varied from 25-49 days. Conclusions: Over the last few decades many dams have been built
in Brazil, since the energetic matrix of the country is based on hydroelectric power stations.
However getting a reasonable equilibrium among the necessary energy for development and
environment and human health is a major challange. The studies here presented are part
of the recommended OMS, SVS/MS and PIDE preventive measures against schistosomiasis
spread throughout man-made lakes and irrigation impoundments. Concerning B. straminea
the results, associated with marked social and ecological changes occurred, strongly suggest
the possibility of B. straminea coming to act as a vector of schistosomiasis if no surveillance
is implemented in the studied areas.
91
PERSPECtIvES Of USIng EuPHORBIA MIlII vaR. HISlOPII tO COntROL IntERMEdIatE
HOStS Of SCHISTOSOMA SPP.
virgínia torres Schall
Laboratório de Educação em Saúde e Ambiente - Centro de Pesquisas René Rachou, Fiocruz,
Minas Gerais
3
Introduction. Schistosomiasis is a neglected human disease that spreads related to several
factors and associated with high levels of poverty. One alternative for its effective control
(integrated with others) is the use of plants (and/or active components) against intermediate
hosts of Schistosoma spp. Currently, there has been a growing interest in the development
of appropriate technologies that would allow the affected communities the use of natural
products with moluscicides properties. This alternative strategy is emphasized in view of the
high cost of synthetic moluscicides, its toxicity to the flora and fauna, and some operational
difficulties in transport and application, which makes them prohibitive for this purpose.
The plant Euphorbia milii (syn. splendens) var. hislopii, whose property was identified as a
molluscicide by Vasconcellos & Schall (1986), has a low lethal concentration and meet the
first requirement posed by the World Health Organization (WHO), which provides up to 20
ppm, the lethal dose (LD 90) for a plant be considered effective. As from its discovery, this
plant has been investigated, and the main results obtained so far will be presented here.
Methodology. The species Euphorbia milii (Des Moul) (Syn. splendens), var. hislopii is
originated from Madagascar, and is used in Brazil as ornamental hedge. It is also known as
“crown of Christ”, “two friends” and “mattress of bride”. It can be easily grown in endemic
areas and produces large amount of latex throughout the year. By means of the standardized
methodology of the WHO for testing plant-derived molluscicides, it was investigated LD90
for intermediate hosts of schistosomiasis in Brazil and Africa, exposed to E. milii lyophilized
and natural latex. Field studies were also conducted in experimental streams located in
endemic areas. Studies are under way to determine the viability of obtaining the natural and
lyophilized latex in large scale, in order to assess its use in an integrated control program with
community participation. Results. Laboratory results showed that the DL90 using an aqueous
solution of E. milli var. hislopii latex was lower than 0.5 ppm for B. glabrata and B. tenagophila
created in the laboratory and 4.00 ppm for B. tenagophila brought from the field. In a further
laboratory study, the DL90 ranged from 0.13 ppm for B. glabrata (dried latex) to 4.00 ppm
for B. pfeifferi (latex “in nature” (Schall et al, 1998). The DL90 was also established for the
species B. straminea and Bulinus sp. The lethal action demonstrated for molluscs vectors of
schistosomiasis in Africa (B. pfeifferi and Bulinus sp.) opens perspectives for field studies in
this continent, where schistosomiasis presents high prevalence in several countries. Tests in
the field habitats lotic (B. tenagophila) and lentic (B. glabrata) showed 100% of mortality for
concentrations of 5 and 12 ppm (Mendes et al., 1992; Baptista et al., 1992). Other laboratory
experiments have shown that the latex has seasonal and geographical stability for samples
collected in Brazil and temporal stability after a two-year storage in a refrigerator at 10o C.
(Schall et al., 1992). Later laboratory and field investigations (Vasconcellos, 1996; Vasconcellos
and Amorim, 2003) demonstrated the lethal effect of aqueous solutions of latex of E. milli
for the snail L. columella, that is, the main intermediary host, in Brazil, of F. hepatica, a worm
that causes fasciolosis. Chemical studies allowed to isolate and identify eight miliaminas
92
present in the latex of E. milii, which characterize the moluscicide active principle in latex
(Zani et al., 1993). The latex is less toxic than the niclosamida for fish, microcrustaceans,
oligoqueta, larvae of mosquitoes of first and third stages. Latex did not inhibit the growth
of algae clorofíceas and presented no inhibitory effect for the bacteria. The activity on B.
glabrata was quickly lost when solutions of latex were exposed to visible light (Oliveira Filho
& Paumgartten, 1995). Studies made by Baptista et al. (1994, 1997) estimated the latex cost
and benefit, and indicated the economic and operational viability of the cultivation of the
plant, of the extraction of latex and of its application in large endemic areas. Conclusion.
All the above properties, together with the wide distribution of the plant, its resistance
and adaptation to tropical climate, the facility of its cultivation, the easy obtention of latex
and preparation of the molluscicidal solution, make this a promising product for large-scale
use in the control of schistosomiasis. A viability study was recently concluded and shortly
it will begin a phase of cultivation and latex extraction and lyophilization to use in some
endemic areas, aiming at evaluating the moluscicide effect in the schistosomisis control in
an integrated approach.
Supported by CNPq and Fapemig
93
IMMUnE RESPOnSE Of PatIEntS RESIStant tO InfECtIOn BY SCHISTOSOMA MAnSOnI
Ricardo R. Oliveiraa,c, Joanemile P. figueiredoa, Luciana S. Cardosoa,c,e, Rafael L. Jabara, Robson
P. Souzaa, Martin t. Wellsd, Edgar M. Carvalhoa,b,c, daniel W. fitzgeraldf, Kathleen C. Barnesh,
Maria Ilma araujoa,b,c, Marshall J. glesbyg
Serviço de Imunologia, Hospital Universitário Prof Edgard Santos, Universidade Federal da
Bahia, Salvador, Bahia, Brazil; bEscola Bahiana de Medicina e Saúde Pública, Salvador, Bahia,
Brazil; cInstituto Nacional de Ciência e Tecnologia de Doenças Tropicais (INCT-DT/CNPq),
Salvador, Bahia, Brazil; dDepartment of Biological Statistics and Computational Biology,
Cornell University, Ithaca, New York, United States of America; eDepartamento de Ciências
da Vida, Universidade do Estado da Bahia, Salvador, Bahia, Brazil; fCenter for Global Health,
Division of Infectious Diseases, Department of Medicine, Weill Cornell Medical College, New
York, New York, United States of America; gDivision of Infectious Diseases, Department of
Medicine, Weill Cornell Medical College, New York, New York, United States of America;
h
Division of Allergy and Clinical Immunology, Department of Medicine, Johns Hopkins
University, Baltimore, Maryland, United States of America
a
Introduction: Subjects with naturally or post-treatment acquired resistance to infection
have been identified in endemic areas of schistosomiasis. This resistance to infection seems
to be mediated by the parasite-specific immune response. Methodology: We conducted
a cross-sectional study of subjects resistant to schistosomiasis and matched controls in an
endemic area in Bahia-Brazil. Blood was collected to obtain serum for antibody evaluation
by ELISA. Peripheral blood mononuclear cells (PBMCs) were stimulated in vitro with the
S. mansoni antigens SWAP, SEA, Sm21.6, Sm22.6, PIII or IPSE and levels of cytokines were
determined by ELISA. Results: Socio-economic variables were not associated with resistance
to infection with S. mansoni. PBMCs of resistant subjects produced higher levels of IL-5,
IL-13 and IFNγ upon stimulation with SEA, compared to infected individuals. On the other
hand, the Sm21.6, Sm22.6, PIII and IPSE antigens induced significantly lower production of
IFNγ by cells of resistant individuals as compared to infected individuals. The Sm21.6 and
Sm22.6 antigens induced higher levels of IL-10 in resistant subjects than in susceptible ones.
Ratios between IL-5, IL-10 and IFNγ showed a co-existence of both Th1 and Th2 predominant
immune response in the resistant group. Unexpectedly, the levels of IgE specific to SWAP
and IPSE were lower in the resistant group. However, the levels of IgG4 specific to SWAP,
SEA and Sm22.6 were also lower in the resistant subjects compared to the susceptible ones.
Conclusion: Our data suggest that the socio-economic variables examined could not fully
explain resistance to S. mansoni infection observed in the studied area. However, a mix of
both Th1 and Th2 immune response together with low levels of specific IgG4 for parasite
antigens could be mediating the resistance to infection
94
InnatE and adaPtIvE IMMUnE RESPOnSE In HUMan SCHIStOSOMIaSIS ManSOnI
andréa teixeira-Carvalhoa*, Cristina toscano fonsecab, Renato abdalaa, amanda Cardoso de
Oliveira Silveiraa, Olindo assis Martins-filhoa, andréa gazzinellic, Sérgio Costa de Oliveirad,
Egard Marcelino de Carvalho filhoe, Rodrigo Corrêa-Oliveiraf
Laboratório de Biomarcadores de Diagnóstico e Monitoração, Centro de Pesquisas René
Rachou, Fiocruz, Belo Horizonte, Brasil; bLaboratório de Esquistossomose, Centro de
Pesquisas René Rachou, Belo Horizonte, Brasil; cEscola de Enfermagem, Universidade Federal
de Minas Gerais, Belo Horizonte, Brasil; dLaboratório de Imunologia de Doenças Infecciosas,
Universidade Federal de Minas Gerais, Belo Horizonte, Brasil; eServiço de Imunologia,
Universidade Federal da Bahia, Hospital Universitário Professor Edgard Santos, Salvador,
Bahia, Brasil; aLaboratório de Imunologia Celular e Molecular, Centro de Pesquisas René
Rachou, Fiocruz, Belo Horizonte, Brasil;
a
Introduction: Previous studies peformed by our group have demonstrated that distinct
immune response profiles can be correlated with the development/maintenance of
different clinical forms of human schistosomiasis. Moreover, several individuals who
have always lived in endemic areas for schistosomiasis stay protected from infection.
An important question for disease control in developing countries is which differences
in the immunological profile of these negatively tested (non-infected) individuals can
account for their resistance to either infection or reinfection. Moreover, complex
immune mechanisms lead to the slow acquisition of immune resistance. In all these
cases, innate immune factors also play a part. Methodology: This work is a crosssectional study of a population living in a rural community, São Pedro, in the state
of Minas Gerais, Brazil. Eighty volunteers participated of the study (40 individuals
without and 40 individulas with Schistosoma mansoni infection detected by KatoKatz test, Sch- and Sch+ groups, respectively). To better understand the immune
mechanisms associated with the schistosomiasis control, we present the results of the
ex vivo evaluation based on the frequency of the natural killer (NK) cells expressing
activation markers (NKG2D, NKp46 and NKp80) and dendritic cells expressing Tolllike receptors (TLR)-1, -2 and - 3 by flow cytometry. We have also performed an ex
vivo frequency of putative regulatory T-cell (Treg) and B-cell (Breg) subsets in the
peripheral blood mononuclear cells from individuals living in S. mansoni endemic
area. In parallel, we have carried out the analysis of cell proliferation after in vitro
stimulation from peripheral blood mononuclear cells of these individuals with soluble
eggs/adult worms S. mansoni antigens. Results: We observed an increase on NKp80+
and NKG2D+ in the NK cells in Sch+ patients and a significant decrease on the frequency
of Treg+ (iCTLA-4+ CD25+ CD4+) cells as well as Breg+ (CD40+CD1d+CD5+CD19+)
cells in Sch+ patients as compared to Sch- individuals. Correlation analysis showed
a positive relation between these two regulatory subpopulations. We have also
observed an increased frequency of TLR-2+ dendritic cells in Sch+ patients. Analysis
of cell proliferation pointed out an predominance of CD4+ cells to proliferate in the
presence of soluble S. mansoni antigens in comparison to CD8+ cell subsets in both
Sch- and Sch+ groups. Conclusion: Innate immune cells as NK and dendritic cells may
play a role in the activation process occurring during human S. mansoni infection and
95
probably additional immunoregulatory events did not triggered by Treg and Breg cells
are important for disease control.
Financial support: Instituto Nacional de Ciência e Tecnologia de Doenças Tropicais
(INCT-DT)/CNPq, FIOCRUZ and FAPEMIG.
96
IndUCtIOn and REgULatIOn Of tHE PatHOgEnIC tH17 RESPOnSE In SCHIStOSOMIaSIS
Miguel J. Stadecker, Laura I. Rutitzky, Mara g. Shainheit, Patrick M. Smith
Department of Pathology, Tufts University School of Medicine, Boston, Massachusetts, USA
Both in humans as well as in mice, infection with Schistosoma mansoni can develop into
severe and mild forms of disease. Naturally occurring severe disease observed in CBA
mice is characterized by marked egg-induced hepatic granulomatous inflammation and
robust production of Th17- and Th1-associated cytokines, whereas in BL/6 mice, milder
immunopathology correlates with a largely Th2-biased cytokine response. In BL/6 mice,
pronounced exacerbation of immunopathology induced by immunization with schistosome
egg antigens (SEA) in CFA (SEA/CFA) substantially recapitulates the natural high pathology
seen in CBA mice; both are associated with a significant elevation of IL-17 and IFN-γ. We
now investigated the relative contribution of IL-17 and IFN-γ in pathology exacerbation of 7
week-infected, SEA/CFA-immunized, IL-17-/-, IFN-γ-/-, and novel double IL-17/IFN-γ-/- BL/6
mice. IL-17-/- mice developed relatively mild egg-induced immunopathology together with
an increase in IFN-γ production by SEA-stimulated mesenteric lymph node and granuloma
cells in vitro, whereas in IFN-γ-/- mice a pronounced exacerbation of immunopathology
was associated with increased IL-17. In IL-17/IFN-γ-/- mice, complete resistance to SEA/
CFA-induced disease exacerbation was associated with a reduction in IL-23, IL-1β and the
chemokines CXCL1 and CXCL2, and with an increase in IL-5 and IL-10. We found that in
wild-type mice IL-17 and IFN-γ are derived from distinct CD4 T cells in which production
of each of these cytokines is suppressed by the other. These results demonstrate that
marked disease exacerbation in schistosomiasis is mainly driven by IL-17 and regulated by
IFN-γ; however, in the absence of IL-17, IFN-γ is capable of exerting a limited, yet significant,
pathogenic function. We also examined the requirements for Th17 cell development in
high-pathology CBA mice using novel CD4 T cells expressing a transgenic (Tg) TCR specific
for the major Sm-p40 schistosome egg Ag, which produce IL-17 when stimulated with the
I-Ak-restricted immunodominant peptide 234-246 (Sm-p40234-246). The ability of CBA bone
marrow-derived dendritic cells (DC) plus Sm-p40234-246 peptide or live schistosome eggs to
stimulate IL-17 production by Tg T cells in vitro was inhibited by blockage of IL-23 and IL-1 β,
but not IL-6; conversely, exogenous addition of IL-23 and IL-1 β effectively reconstituted IL-17
production by egg-stimulated Tg T cells cultured in the presence of syngeneic CBA DC lacking
IL-12p40, the common subunit of IL-23 and IL-12. Kinetic analysis demonstrated that the
IL-17 response was initiated by IL-23, and significantly strengthened by IL-1 β. Importantly,
schistosome-infected IL-12p40-/- CBA mice, or CBA mice treated with IL-1R antagonist,
developed significantly milder hepatic immunopathology together with a dampened egg Agspecific IL-17 response. These findings identify IL-23 and IL-1 β as critical cytokines in the
differentiation of Th17 cells that mediate severe schistosomiasis.
Supported by the US Public Health Service.
97
QtL MaPPIng Of a LOCUS COntaInIng gEnES fOR RESIStanCE tO OxaMnIQUInE
Claudia L. valentim; Claudia C, Queiroz; Matthew Berriman; Jason tsai; timothy JC anderson;
Philip t. Loverde
Departments of Biochemistry and Pathology, University of Texas Health Science Center
Introduction: Schistosomes show heritable variation in biomedically important traits such as
drug resistance, host specificity, and virulence. Our central aim is to develop forward genetic
methods (i.e. linkage mapping) to identify parasite genes that underlie this phenotypic
variation. This approach is well suited to S. mansoni as the lifecycle can be maintained in
the laboratory and clonal propagation of parasites within snails generates large numbers
of genetically identical parasites. To provide proof-of-principal that this approach is feasible
and powerful, we have mapped a genome region that underlies resistance to oxamniquine
(OXA). Resistance to OXA has arisen in nature, has a simple recessive basis and results in
~500-fold reduction in drug sensitivity. Methodology: We crossed resistant and sensitive
parasites (parents) and then crossed two F1 individuals to generate multiple F2 progeny, at
each stage isolating individual parasite genotypes by infecting snails with single miracidia. We
measured OXA-resistance by monitoring death of cultured worms following drug exposure
and genotyped parents, F1 and F2 progeny using 64 microsatellite markers distributed at
~20cM intervals across the genome. Results: As expected trait segregation in the cross was
consistent with recessive inheritance as F1s were sensitive and ~25% of F2 progeny were
resistant. We used the S. mansoni linkage map developed by our group to locate quantitative
trait loci (QTL) underlying OXA resistance. We found a strong QTL (LOD = 13.5) on the p
arm of chromosome 6 where microsatellite markers segregate closely with OXA resistance.
The two parents of this cross have now been sequenced, simplifying fine mapping and
identification of candidate genes. Conclusions: Successful identification of gene(s) that
underlie OXA-resistance will provide insights into mode of drug action, allow development
of modified compounds that kill resistant parasites, generate selectable markers for genetic
manipulation, and set the stage for forward genetic analyses of a range of biomedically
important traits including praziquantel resistance.
98
StEWaRdSHIP fOR RESEaRCH On InfECtIOUS dISEaSES Of POvERtY and tHE ROLE
Of IntERnatIOnaL RESEaRCH REfEREnCE gROUPS
arve Lee Willingham
WHO/TDR, Geneva, Switzerland
TDR is currently implementing its new ten-year vision and strategy which aims at fostering
effective global research efforts on infectious diseases of poverty in which disease endemic
countries play a pivotal role. In order to achieve the new vision, TDR is using a three-pronged
strategy to: 1) provide a collaborative framework and information service for research
partners through stewardship; 2) empower scientists from disease endemic countries as
research leaders; and 3) support research on neglected priority needs. Under the stewardship
function TDR will play a major new role as facilitator and information provider to support
needs assessment, priority setting and progress analysis, and to provide a neutral platform for
partners to discuss and harmonize their activities regarding infectious diseases of poverty. To
implement this strategy TDR’s Stewardship Team has created an independent “Think Tank”
of 125 internationally recognized experts to serve as an analysis forum for identifying control
challenges, major research gaps and opportunities and to ensure that infectious disease
research priorities are based on evidence and integrated with control needs which highlights
participation and pivotal roles for disease endemic country institutions and decision makers.
The expert “Think Tank” group has been organized into ten Disease-specific and Thematic
Reference Groups (DRGs/TRGs) to identify the top priorities for research on infectious
diseases of poverty (http://apps.who.int/tdr/svc/stewardship/research-think-tank).
One of the DRGs is addressing control challenges and research gaps in order to identify research
priority needs for helminth infections of public health importance including schistosomiasis,
onchocerciasis, lymphatic filariasis, soil transmitted helminthiases, foodborne trematode
diseases and taeniasis/cysticercosis. The DRG on Helminth Infections is being hosted by
the African Programme for Onchocerciasis Control (APOC) based in Burkina Faso where
the DRG held its first meeting in January 2010. Since that time the DRG has conducted
gap identification and analysis based on what is currently known; what available research
is not used or applied; what is not known, and what research is needed. Based on ranking
criteria adopted by all the DRGs/TRGs the DRG on Helminth Infections has identified five
main core themes to address as follows: 1) intervention; 2) epidemiology and surveillance; 3)
environment and social ecology; 4) data and modeling; and 5) biology. The DRG is now in the
process of finalising an initial report identifying priority research needs to meet challenges
in the control of helminth diseases. The reference group report will be used as a technical
document for production of the 2011 issue of the Global Report for Research on Infectious
Diseases of Poverty which will serve as an authoritative and innovative reference, primarily
for policy makers, research funders and the research community. The Global Report will
provide a concise resource of evidence and information to guide decisions on agenda setting
and funding; helping to steer increased resources to priority research areas, overcome
current disparities and fragmentation and encourage utilisation of research results for policy.
The roundtable session provides opportunity to inform further about the “Think Tank”
approach for research for infectious diseases of poverty, the challenges and gaps in
knowledge and top level priority research areas identified by the DRG on Helminth Infections
99
as being crucial for sustainable control, and seek inputs and contributions from stakeholders
attending the ISS. In addition presentations will be given by a variety of stakeholders regarding
research and control needs with regard to the different regions as well as broader issues such
as research addressing operational and implementation needs (e.g. integrated helminth/
NTD control). Following the presentations stakeholders will be given the opportunity to
comment on the draft recommendations of the DRG on Helminth Infections. In this way the
DRG on Helminth Infections will be able to take into consideration suggestions and concerns
emanating from discussions with stakeholders during their DRG meeting aimed at finalising
their report being held immediately after the International Schistosomiasis Symposium.
Objectives:
1. Present the “Think Tank” framework for priority setting for research on infectious
diseases of poverty.
2. Share the Disease-specific Reference Group on Helminth Infection’s preliminary
assessments and recommendations of top level research priority areas to meet the
challenges and control of helminth infections.
3. Seek stakeholders’ inputs into the finalization of the report of the Disease-specific
Reference Group on Helminth Infections.
100
an IntEgRatEd StRatEgY fOR tRanSMISSIOn COntROL Of SCHIStOSOMIaSIS In
CHIna
guo-Jing Yanga, xiao-nong zhoub
a. Jiangsu Institute of Parasitic Diseases, Wuxi, China
b. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention,
Shanghai, China
Background: Despite sustained efforts for its control made over the past 50+ years, the reemergence of schistosomiasis in China was noted around the turn of the new millennium.
an integrated strategy: Consequently, a new integrated strategy was proposed to inhibit
the contamination of schistosome eggs to the environment where Oncomelania hupensis
snail infested. The integrated strategy emphasizes on approaches consisting of health
education, access to clean water and adequate sanitation, mechanization of agriculture and
fencing of water buffaloes, along with chemotherapy. Results: Validation of this integrated
control strategy in four pilot counties in the provinces of Anhui, Hubei, Hunan and Jiangxi
revealed significant reductions in the rate of Schistosoma japonicum infection in humans and
intermediate host snails. The already low human infection prevalence observed in Jinxian
in 2007 (0.5%) further decreased in 2008 to a level as low as 0.08%. In the other pilot sites,
after three or four transmission seasons, the human infection prevalence had dropped from
11.2% to 0.9% in Anxian, from 13.3% to 1.8% in Hanchuan and from 4.7% to 1.5% in Guichi.
Importantly, this strategy showed an impact on diseases beyond schistosomiasis, signified by
concomitant reductions in the prevalence of soil-transmitted helminth infections. discussion:
In view of China’s new integrated strategy for transmission control of schistosomiasis showing
an ancillary benefit on other helminthic diseases, we encourage others to investigate the
scope and limits of integrated control of neglected tropical diseases. The implementation
of such integrated programs in mountainous areas also yielded great reward, i.e. in 2008
transmission of control of S. japonicum has been widely achieved in Sichuan Province, a
mountainous endemic region, marking a milestone in the history of schistosomiasis control
in China. An ambitious proposal has been laid out – to eliminate transmission of the parasite
in Sichuan and Yunnan provinces, by 2015, and to expand the success to the rest of country
to achieve transmission control at the same year.
101
102
EvaLUatIOn Of tHE IMPLEMEntatIOn SCHIStOSOMIaSIS ManSOnI EPIdEMIOLOgICaL
SURvEILLanCE: a StUdY CaSE In tHE MUnICIPaLItY Of UnIãO dOS PaLMaRES In
aLagOaS StatE.
Jeann Marie Rocha Marcelino; Luciano Medeiros de toledo; Helia Kawa; Marly Marques da Cruz
Ministério da Saúde/SVS, DF, Brasil
Introduction: This study aimed to evaluate the degree of implementation of the Surveillance
and Epidemiological control of schistosomiasis, through a case study in the city of União
dos Palmares, Alagoas. In endemic areas the actions of Epidemiological Surveillance of
schistosomiasis are aimed at reducing the occurrence of severe forms, the prevalence of
infection and indicate measures to reduce the risk of spread of the disease using as a main
control strategies for early detection and timely treatment of patients with Schistosoma
mansoni . As additional measures are recommended control of intermediate hosts, the
actions of health education and an indication of priority areas for interventions in the
environment. Methodology: An assessment of the normative components of structure and
process, considering the external environments, organizational and political deployment
of the shares. The study design used a combination of concepts, methods and sources for
collecting secondary and primary data. Initially, the document analysis and consulting to
data from Information System Program Monitoring and Control of Schistosomiasis, System
for Notifiable Diseases, the Hospital Information System and Mortality Information System.
Interviews were conducted with key informants who work in the surveillance and control
of schistosomiasis in the state and municipal level and direct observation of organizational
infrastructure and field work. It was constructed the logical model evaluation and set the
dimensions to be used. The process of building the array of trial with the participation of
users interested in assessing the definition of categories, indicators and evaluation criteria.
The quality of stock was assessed by considering the extent of compliance defined as the
adequacy of actions to the standards prescribed by the Ministry of Health. Results: The
results of this study showed a partial development with external environment (53.7%),
evidenced by the environmental characteristics and socioeconomic characteristics that
favor the occurrence and persistence of schistosomiasis. With regard to organizational and
political context was obtained a ratio of 45.6% and thus classified as partially implemented,
mainly due to problems related to the ability to hire human resources, difficulties in
managing the program and the incipient realization of inter. The context of implementation
of actions in the county was considered partially implemented (64.6%), whose most critical
aspects refer mainly to insufficient human resources to act in controlling the disease, lack
of training and updating of professionals, and difficulties operational activities of active
search and treatment, the latter which was below the goal set by the technical rules.
Conclusions: The municipality performs the actions of epidemiological surveillance and
control of schistosomiasis within its capabilities and limitations, giving priority to cities
with prevalence above 10%. However, it was observed that the surveillance commits
managerial failures in carrying out activities related to systematic monitoring, processing,
analysis, interpretation, recommendations and information dissemination in order to
enhance the action of disease control at the local level. Regarding the activities of active
search and treatment of patients with S. mansoni are performed mostly by agents of
103
endemic Funasa experienced from and by community health workers. We highlight the
importance of the work of these professionals who work with dedication, but in poor
conditions. However, this performance is an isolated and decoupled from other local
health professionals, including the Epidemiological Surveillance. The evaluation model,
considering the limitations, proved to be sufficient to meet the proposed objectives, and
may contribute to the improvement and adjustment of actions developed by the program
in the county.
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PattERn and IMPaCt Of HELMIntH InfECtIOnS In PREgnanCY: PROSPECtS fOR
IMPROvEMEnt MatERnaL HEaLtH In angOLa
Ilda Rosalina Jeremias; Silvana Maria duarte Belo; Joltim Quivinja; filomena gomes da Silva;
Eugénia Ramos; Maria amélia afonso grácio
Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Portugal
Introduction: Improving maternal health in Angola is a priority of national health authorities
as the country has among the highest maternal mortality rates in the world. Various
studies demonstrate that anemia and iron-deficiency anemia may contribute directly to up
20% of worldwide maternal deaths (WHO, 2004) and helminthic infections could strongly
contribute to this situation. The assessment of helminthiasis in pregnant women is not
routinely used in antenatal health clinics in Angola, thus any existing infection will remain
untreated which may cause various adverse effects during pregnancy. This study aims at
evaluating the prevalence of helminthic infections and the influence of helminthiasis on
anemia and iron status in pregnant women both in urban and in rural settings, in order
to promote awareness of these pathological conditions as well as to their preventive
measures. Methodology: A cross-sectional study targeting major infections in pregnancy,
including helminthiasis, is underway at the antenatal heath cares in Angola. Three hundred
and seven (307) pregnant women from the cities of Luanda (country´s capital) and Malange
(central north province) antenatal units, aged from 14 to 48 years old (median 28± 6.69
yrs) were until now enrolled in the study. Haematological (haemoglobin and iron indexes)
and parasitological (urine and faeces) analysis were performed and a questionnaire was
applied to identify potential social and/or behavioural risk factors contributing to any
pathological conditions. Statistical procedures were carried out using statistical package
SPSS 17.0 for Windows. A P-value <0.05 was considered significant. Results: Out of the
307 women screened, 137 fulfils the defined inclusion criteria for helminthiasis-related
to anemia. The prevalence of anemia (Hb<11g/dl) and iron-deficiency (FRT<50 ng/
ml) was 44.5 % and 55.8 % respectively. Lower haemoglobin (Hb) and iron levels were
more frequent in younger women as well as in primigravidae than in multigravidae,
suggesting that women start pregnancy with depletion of iron stores. Schistosomiasis
caused by Schistosoma haematobium and geohelminthiasis due to Ascaris lumbricoides
and Trichuris trichiura were the main helminthiasis found in the urine and faecal samples,
with prevalence of 16,5% and 34% respectively. Intensities of both schistosome and soiltransmitted helminthes were significantly associated with low haemoglobin and iron
levels (Spearman’s rho <0.05), indicating their potential role on inducing iron deficiency.
Comparing the two regions, helminth infections were predominant in women from
Malange, particularly for S. haematobium and were more frequent on larger households
(more than 4 children). In addition, their frequency and intensity were positively associated
with low economic income and lack of water supply seems to be the main risk factor for
helminthiasis, in particular for women living in suburban areas. (OR=3.09, 95% CI: 1.548.32, P=0.024). The schooling level of women (secondary) and the previous knowledge
of anemia and helminthiasis, were facts that did not accompanied their awareness for
treatment and preventive measures. Conclusions: The findings of this study indicate that
helminthiasis and iron deficiency anaemia could be high prevalent not only in this particular
105
group but one could generalise these data to women of reproductive age and eventually
other population’s group. Programs should include the control of helminthic infections in
routine antenatal cares as anthelminthic treatment reduces the health risks associated to
low hemoglobin levels and could potentially improve both mother and child health.
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SURvEY Of SOCIO EPIdEMIOLOgICaL data On PatIEntS WItH SCHIStOSOMIaSIS Of
CaPELa dO SOCORRO, SOUtH REgIOn Of SãO PaULO, In tHE PERIOd Of 2004 tO 2010.
Beatriz aparecida Imparato; andréia aparecida Caggegi; Rafael de Oliveira Christe; nathália
Rafaella Martinho; Elaine Cristina S gama; Léia Munhoz Parra; ana Lúcia de Lima gabriel;
Patricia Placoná diniz
Prefeitura Municipal de São Paulo, Brasil
Introduction: The World Health Organization points out a group of infectious diseases
considered neglected due to the low investment in researh of control and treatment. These
are diseases that occur mainly in tropical countries with low income, being promoted in
poverty and poor sanitation. Among them is schistosomiasis, which can lead to a chronic
infection resulting in long periods of disability with high impact on economic development
and education of populations. Schistosomiasis is present in over 76 countries. In Brazil more
than 25 million people live in endemic areas and 2.5 million have symptoms. In general,
states with higher occurrence are Bahia and Minas Gerais and others of northeast region.
The Capela do Socorro is located in the southern city of São Paulo, being divided into three
administrative districts: Grajaú, Cidade Dutra and Socorro. The area is characterized by
having two dams in the territory: Billings and Guarapiranga being of extreme importance as a
source of recreation and water supply for the city. With an area of 134.2 km2 the population
density is around 684.000 people, of which 65% are in Grajau, 29.7% in the Cidade Dutra
and 5.3% in Socorro. Methodology: To survey the socio epidemiological data of the patients
with schistosomiasis in the region were assessed compulsory reporting forms passed to the
Supervision of Health Surveillance (SUVIS) of Capela do Socorro after attendance in the Basic
Health Units. We evaluated the records of the period between 2004 and August 2010. They
were then carried out home visits to the patients and a questionnaire was aplied to determine
the socio epidemiological datas, the suspected infection site, how much the patients knew
about the disease and prevention methods. In the visits were also analyzed the perimeters
of homes to check the conditions of sanitation and the potential disperser of the disease
in the region, linking the presence of streams, dams and other water collections. Results: A
Total of 120 cases were reported to SUVIS Capela do Socorro, where 72.5% were in Grajaú,
23.3% in Cidade Dutra and 4.2% in Socorro, a district with higher infrastructure and income.
Of the 120 cases reported were carried out 49 interviews, 20 patients were not located
(address and phone does not ring true). Only one case was considered as autochthonous
and one another still listed as inconclusive. 52% of patients live in areas with deficiencies
in basic sanitation, 84% live near streams and/or dams, 58% have attended or still attend
various bodies of water in the region; 59.5% were asymptomatic, being diagnosed with the
disease in tests routine, 40.5% had symptoms; 12.5% were discovered in the Pre-Natal tests
and 6.25% were detected because of other diseases. Of the patients with schistosomiasis,
90% are not natural state of São Paulo, 61% migrated in search of better living conditions,
34% migrated to follow the family, and 4.5% to study. Likely sites of infection have shown a
prevalence of the states of Bahia, Minas Gerais and Pernambuco. Regarding awareness of the
patients, 67% knew how to infection and 77% did not know how to prevent it. Conclusions:
It is expected that only a proportion of cases of schistosomiasis is notified. Most patients are
asymptomatic, and those showing symptoms often do not search a health unit, and even
107
when detected the disease does not perform the treatment or return to check the healing.
Patients most often had no access to information in relation to schistosomiasis, showing a
deficiency of service by health units. The Capela do Socorro has great potential disperser of
the disease since the Grajaú, which has the highest incidence of cases, has only 39% of the
sewage collection, with most of its waste released into streams that flow into the dam or up
directly in the water collections that people often use as a source of economic and leisure.
Corroborating to the disperser potential of schistosomiasis in the region, scientific studies
showed the presence of intermediate hosts in the two dams. In this context it is necessary to
carry out an educational project involving health professionals, education professionals and
populations at risk, raising awareness of the severity of the disease.
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COnStRUCtIOn Of COMPUtERIzEd tEMPLatES fOR EPIdEMIOLOgICaL SURvEYS On
SCHIStOSOMIaSIS: ExPERIEnCE In PORtO dE gaLInHaS, PERnaMBUCO.
Constança Simões Barbosa; Elainne Christine Souza gomes; Onicio Batista Leal neto
Centro de Pesquisas Aggeu Magalhães, Fiocruz, PE, Brasil
Introduction: On the coast of the state of Pernambuco and the metropolitan region of Recife,
schistosomiasis has become established in focal form with seasonal transmission. Lack of
sanitation, water supply and healthcare information have contributed towards systematic
fecal pollution in coastal freshwater aquatic environments, thereby maintaining disease
transmission in a region that is tending to become endemic for the disease, because of the
specific characteristics of the spatial occupation patterns. In the district of Vila de Porto de
Galinhas, on the southern coast of Pernambuco, large numbers of foci of vector molluscs
for schistosomiasis have been recorded since the flooding that occurred in 2000. That event
induced an outbreak of acute schistosomiasis in 662 individuals who were accidentally
exposed to water full of infected snails. In June 2010, new floods ravaged the municipality
and the community of Vila de Porto de Galinhas was again submerged and exposed to
infected snails. In July 2010, the Schistosomiasis Laboratory of CPqAM/Fiocruz started an
epidemiological mapping survey with the aims of updating the data gathered in the year
2000, identifying new cases of the disease and investigating the environmental and biological
factors that persist there and are determinants of seasonal production of acute human cases
of this disease. Methodology: Field epidemiological investigations start with geographical
recognition, which takes the form of a computerized template in which the components
of an epidemiological unit (locality) are described through geometric tracings, graphs and
statistical data. During the months of June, July and August 2010, a georeferenced template
of the locality was constructed in order to define the limits of the area to be investigated and
its local physical attributes (homes, street blocks, breeding sites and geographic features).
These data were georeferenced using a GPS (Global Positioning System) device: eTrex, on
the Garmin platform. To mark out homes and commercial establishments, the methodology
indicated in the geographic recognition manual of the National Health Foundation (FUNASA)
was used. The homes and commercial establishments were numbered sequentially, from
no. 1 upwards to infinity, in an “S” pattern, going from left to right and returning from right
to left in the second line of street blocks, in a sequence such that each street block was as
close as possible to the next one, thereby avoiding large shifts. All the data stored in the
field using the GPS device were transferred via the universal serial port (USB) to a computer,
in which the data were interfaced using the GPS Track Maker (GTM)® software in order to
adjust the template and make corrections to the spectrum of GPS precision (which is 25
meters). To finalize the template, the Corel Draw 13 software was used to make esthetic
adjustments to the map and include geographic elements such as the points of the compass
and scale bars. Results: In the georeferenced template for Vila de Porto de Galinhas, all the
homes can be seen, according to neighborhoods and street blocks (Fig. 1). The Corel Draw 13
software make it possible to add the distribution of the Biomphalaria glabrata breeding sites
so far detected, to the template in the form of symbols (Fig. 2). Addition of complementary
data to the template, resulting from parasitological and malacological surveys that are in
progress will yield consistent epidemiological information in the form of thematic maps on
109
which the distribution of human cases positive for S. mansoni and the foci of vector snails
will be displayed. Another spatial analysis using the resources of the TerraView and ArcView
software will then be made in order to spatially organize the risks within these thematic
maps, from gradation of the human parasitic loads diagnosed and the Biomphalaria
infection rates. Conclusions: Computerized templates detail and position biological and/or
environmental information of research interest within the space under investigation, thereby
rapidly generating epidemiological maps of risks that are valuable instruments for planning
the operational strategies of the schistosomiasis control program and for the territorial
organization actions of the family health program. In addition, the templates can be exported
to software such as TerraView, for statistical and geospatial analysis with more refinement
regarding events that have occurred at the locality.
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EvaLUatIOn Of PaRaSItOLOgICaL dIagnOStIC MEtHOdS fOR SCHIStOSOMIaSIS
ManSOnI In LOW tRanSMISSIOn aREa In MInaS gERaIS, BRazIL
Liliane Maria vidal Siqueira; Áureo almeida de Oliveira; Cristiano Lara Massara; nidia
francisca de figueiredo Carneiro; Paulo Marcos zech Coelho; Martin Johannes Enk
Centro de Pesquisas René Rachou, Fiocruz, MG, Brasil
Introduction: It is scientifically well documented that the prevalence of schistosomiasis
mansoni has been underestimated, since the diagnostic methods currently available lack
sensitivity in estimating the prevalence of the disease in areas of low transmission or in
individual cases with low worm burden. Various studies have been carried out in order to
develop more sensitive and effective techniques for stool examination. Methodology: This
study was conducted in Pedra Preta, located in the rural region of Montes Claros, Minas
Gerais, Brazil and endemic for schistosomiasis. The parasitological survey carried out among
the 218 participants involved three different techniques: the Kato-Katz method, the Three
Fecal Test (TF-test®), and Saline Gradient technique. Four fecal samples per individual were
collected in adequately identified plastic containers during four consecutive days. In case of
the Kato-Katz method 12 slides of the first sample, 2 slides of the second, third and fourth
were prepared, resulting in a total of 18 slides per individual. In case of TF-test®, 500 mg of
the three first stool samples were collected and stored in tubes containing a preservative
solution and afterwards processed according to the protocol. In case of the Saline Gradient
Method, an amount of 500 mg of feces derived from the first sample was diluted in 3.0 mL of
0.85% saline, and processed under a continuous flux of saline solution, the sediment obtained
was preserved in 10% formalin, and examined later. Results: A total of 63 (28.9%) participants
tested positive for S. mansoni considering all slides analyzed by the Kato-Katz method, 56
(25.7%) were found to be positive for hookworm, 10 (4.6%) for Enterobius vermicularis,
and 3 (1.4%) for Trichuris trichiura infections. The results of the TF-test® showed 33 (15.1%)
positive participants for S. mansoni, of which 14 were not detected by the Kato-Katz method
and 7 (3.2%) participants tested positive for hookworm. Using the Saline Gradient Method, 13
(5.9%) participants were found positive for S. mansoni, but one of them was not detected by
the other techniques. Five (2.3%) participants presented a positive result for hookworms and
2 (0.9%) for E. vermicularis. A possible explanation of this lack of sensibility of this technique
may be the prolonged storage time in formalin solution before examination, causing the eggs
dehydration and decrease of density, consequently eggs loss. Using all three parasitological
methods previously mentioned 78 individuals were positive for S. mansoni. This result
indicates an increase of the prevalence from 7.8%, obtained with one slide according to the
Kato-Katz method, to 35.7%, considering all coproscopical techniques. This 4.6 fold increase
of the number of positives clearly shows the significant underestimation of the diagnostic
approach currently in use for areas of low prevalence and infections with low worm burden.
In relations to individual infection intensity, eggs per gram (epg), the TF-test® and the Saline
Gradient technique classified all 34 positives detected as low infection intensity. The Kato
Katz method using 18 slides in 4 samples revealed 61 participants with low epg (1-100) one
with medium (101-400) and one with high (>400), showing 156 and 555 epg, respectively.
The study also demonstrates and confirms data of the literature that the increase of the
number of samples collected on different days is more effective than increase of the number
111
of the slides of the same sample. The results of the increasing number of slides using the
same stool sample revealed 17 (7.8%) positives for one slide, 20 (9.2%) for three, 27 (12.4%)
for six and 31 (14.2%) for 12. In contrast to these findings, 40 (18.3%) positives were detected
by examining three slides of three different stool samples (one in each) and 47 (21.5%) in
six slides of three samples (two in each). Conclusions: The results indicate that prevalence
in areas of low transmission is significantly underestimated, in our case 4.6 times, which has
a negative impact on control strategies based on a single slide of a single sample. Therefore
a more complex diagnostic approach is required under these circumstances, characterized
by very low individual egg count per gram. One of the alternatives to overcome this lack
of sensibility is, as shown here, the increase of stool samples collected on different days.
Another alternative is the combination of different diagnostic methods which also increase
the overall sensibility in order to obtain more reliable values for the prevalence of the
disease. As the profile of schistosomiasis is changing, characterized by an increase of low
transmission areas where the current diagnostic tools cannot provide sufficient sensibility,
control strategies, in order to maintain efficacy, need to be adapted adequately.
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SOCIOEnvIROnMEntaL PROfILE Of PatIEntS WItH SCHIStOSOMIaSIS ManSOnI In
LOW EndEMIC SEttIng In CEaRa StatE.
fernando Schemelzer de Moraes Bezerra; Marta Cristhiany Cunha Pinheiro; Mariana Silva
Sousa; teiliane Rodrigues Carneiro; Sara Menezes de Oliveira; Maria de fátima Oliveira
Universidade Federal do Ceará, CE, Brasil
Introduction: Schistosomiasis mansoni is a parasitic infectious disease caused by digenetic
trematode Schistosoma mansoni. The rates and intensity of infection are influenced within
a population according to their patterns of contact with water, acquired immunity and
behavioral factors, professional, cultural and religious. The development in the area of
water resources has led to creation of reservoirs, dams, and the implementation of irrigation
systems, facts that often lead to expansion of the habitat of the intermediate host and
thus create new potential sites for transmission of schistosomiasis. Current data from the
Schistosomiasis Control Program - PCE, has shown that the State of Ceará is characterized
as an area of low prevalence, but also indicate that the municipality of Maranguape owns
the place with the highest positive rates of schistosomiasis in the state, Planalto do Cajueiro,
where the prevalence of the disease has increased each year. According to recent surveys
conducted in that locale, the prevalence was 8.53% in 2006, rising to 13.76% in 2007.
Therefore, our objective was to provide a description of the socio-environmental community
in Planalto do Cajueiro, Maranguape, relating to positive for schistosomiasis, as determined
by examinations coproscopic. Methodology: There were three stages: home visit to signing
the consent form, interview for collection of epidemiological data by applying a questionnaire
socio-economic and demographic, delivery and collection of pots for completion of stool
examinations in the laboratory. We interviewed 250 individuals and 80 of these gave the
stool sample. Results: The age, 22 (8.8%) were between 2 and 6 years, 18 (7.2%) between
7 and 9 years, 35 (14%) between 10 and 14 years, 57 (22, 8%) between 15 and 25 years, 70
subjects (28%) between 26 and 46 years and 48 (19.2%) with 47 years or more. In making
the crossing between age and positivity, we observed that 2 (6.9%) were positive for 7-9
years, eight (27.6%) 10-14 years; 2 (6.9%) of 15 to 25 years, 11 (37.9%) 26-46 years and six
(20.7%) were older than 47 years. Regarding gender, 100 (40%) were male and 150 (60%)
were female. In making the intersection between gender and positivity, we observed that
15 (51.7%) of those infected were men while 14 (48.3%) were women. Regarding schooling,
26 (10.4%) individuals had completed high school, 28 (11.20%) had finished elementary
school and 16 (6.40%), incomplete secondary education, while 142 (56.8 %) had incomplete
primary education and 38 (15.2%) did not study. The age group 2-6 years was included in the
population study. In making the crossing between schooling and positivity, we observed that
19 (65.5%) infected had incomplete primary education, 6 (20.7%) completed elementary and
4 (13.8%) average incomplete. As for previous infection with S. mansoni, fifty-two patients
(20.8%) said they already had schistosomiasis. By analyzing these data in relation to positive
patients, 23 (79.3%) said they had no previous infection by this disease and 6 (20.7%) said
they had had. As for knowledge about how to get the disease, half , ie 127 (50.8%) said
they knew and 123 (49.2%) not. Similar data were found positive when correlated with the
given above, in which half the number of infected people said they knew how to handle the
disease and half did not. Regarding sanitation, the bathroom was present at the residence of
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237 (94.8%) respondents and piped water in 229 (91.6%). To correlate these data with the
number of positives, we found that 27 (93.1%) had positive bathroom and running water in
their homes. As the profile of water use, 122 (48.8%) residents say they have no contact with
streams running through the community, while 65 (26%) said they use water for recreation
(fishing, swimming, etc.. ), 51 (20.4%) use it for bathing and 34 (13.6%) to perform household
chores (washing clothes and cooking utensils). Conclusions: The community of Planalto do
Cajueiro, Maranguape-Ceará presents a socio environment that is very similar to rural and
semi-urban areas of northeastern Brazil. In this location the continuity of transmission of
schistosomiasis is due to low levels of education of individuals and inadequate sanitary
conditions.
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tHE aPPLICatIOn Of gEOgRaPHIC InfORMatIOn SYStEM (gIS) dURIng a OnE YEaR
OBSERvatIOn PERIOd Of InfECtIOnS WItH SCHISTOSOMA MAnSOnI In PEdRa PREta,
MUnICIPaLItY Of MOntES CLaROS, MInaS gERaIS - BRazIL
Ricardo José de Paula Souza e guimarães; Cristiano Lara Massara; Martin Johannes Enk
Introduction: Schistosomiasis in Brazil is caused by Schistosoma mansoni and the intermediate
hosts are mollusks of the Biomphalaria genus (Mollusca: Pulmonata, Planorbidae). B. glabrata,
B. tenagophila and B. straminea have been found naturally infected by S. mansoni. It is a
parasitosis determined in space and time by environmental and behavioral factors of residents
in endemic areas. The geoprocessing can be applied to characterize and to better understand
the interconnection of these factors. Computational resources, such as GIS, allow a complex
analysis of a large number of information and display the results of this analysis in graphical
maps. Data generated by the GIS have an important role in the study of schistosomiasis,
especially in relation to the interaction of disease with environmental conditions. This work
intends to identify the spatial distribution of patients infected with S. mansoni before and 30,
90, 180, and 360 days after treatment as well as to define areas of disease transmission at the
location. Methodology: The study was carried out in Pedra Preta, a village in the municipality
of Montes Claros, Minas Gerais in the year 2008. The Kato Katz technique was used for the
parasitological survey, collecting a stool sample on four consecutive days and examining 18
slides of each participant. Among the positive participants, 48 completed all sampling stages
before and after treatment. Digitalized vector data were obtained from the municipality of
Montes Claros for the creation of the cartographic data base. A GPS receiver was used for
the spatial location of the participants’ residences and possible breeding sites of Biomphalaria
spp. snails. The GIS was applied for mapping and analyzing data obtained in the field. The
results are presented in the form of thematic maps. Results: A total number of 55 participants
infected with schistosomiasis were identified among the inhabitants of the locality, indicating
a prevalence of 32%. The patients were re-examined 30, 90, 180 and 360 days after treatment
with praziquantel and showed a prevalence of 13%, 2%, 46% and 19%, respectively. Nine of
these positive participants were infected before and 360 days after treatment and one of the
nine 90 days after treatment and five of them 180 days after treatment. All infected patients
are living in a mean distance of 500 meters to the same little pond, where a considerable
number of B. glabrata was found. This little pond is used for recreational activities and its
drainage cuts a road that serves as passage for the residents. During the rainy season this road
is flooded and submerged by water of this pond. Conclusions: The applied technique allows
the spatial configuration of the locations where the largest number of infected individuals was
found. The distance between the residences of positive participants made it possible to guide
the malacological survey into the direction of the possible transmission sites. Due to the high
rate of re-infection in this population it is desirable, in addition to the educational activities
carried out during the field work, to construct a bridge in order to avoid flooding of the passage
with water possibly contaminated with cercariae. The results show that GIS is a useful tool in
the control of schistosomiasis, which can reduce costs and lead the field work, indicating areas
with increased probability of disease occurrence and transmission. Financial support: CPqRR.
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ULtRaStRUCtURaL MORPHOLOgY Of adULt SCHISTOSOMA MAnSOnI, HaRBOREd
In nOn antI-HELMIntHIC tREatEd, HIgH-IMMUnE-tOLEROgEnIC and LOWInfLaMMatORY MICE BY tRanSMISSIOn ELECtROn MICROSCOPY
aurelizia Maria Lemos xavier; antonio Carlos Silva; daniel tavares; Beatriz ferreira Ribeiro;
Susane Borges Rodrigues; Erick vaz guimarães; Luis Cláudio Muniz-Pereira; Maria de fátima
Sarro-Silva; antonio Henrique almeida de Moraes neto
Universidade Fedearl Fluminense, Universidade do Estado do Rio de Janeiro, Instituto
Oswaldo Cruz, Fiocruz, RJ, Brasil
Introduction: In our previous study were observed Schistosoma mansoni tegumental
alterations due to contact with the immune system of non anti-helminthic treated mice. We
compared, by SEM, the tegument of adult worms recovered from strains of mice genetically
selected to extreme phenotypes of resistance (TR strain) and susceptibility (TS strain) to eggalbumin oral tolerance (OT). The parasites recovered from TR mice displayed no morphologic
alteration, while specimens collected from TS mice presented tubercle swelling with blunted
and shortened spines in lower density, increased sensory organelle numbers, fusion and
tegumental ridge peeling. These tegument alterations were similar to those described for
Artemether or Praziquantel treatment, supporting observations that the host immune system
influences the development and function of the tegument of worms harbored in both antihelminthic treated and non-treated mice. Our results were indicative that the development
and function of the worm tegument depend on the immune regulatory capacity of each
individual host. In the present work we observed the inner ultrastructural morphology of
S. mansoni adult by Transmission Electron Microscopy (TEM), obtained from TR and TS
mice, to compare and corroborate our previous Scanning Electron Microscopy (SEM) results
correlated with immune regulatory characteristics of each strain. Methodology: TR and TS
strains of mice from the F25 generation, obtained by two-way genetic selection according
to susceptibility (TS) or resistance (TR) to OVA oral tolerance, were adopted. The original
foundation population, from which the TR and TS strains were derived, was achieved by the
balanced intercrossing of eight inbred mouse strains (Jackson Laboratory, Bar Harbor, ME) of
distant origins (A/J, DBA/2J, P/J, SWR/J, SJL/J, CBA/J, BALB/cJ, and C57BL/6J). The selective
breeding that gave rise to these strains was carried out in the Genetics Department of the Rio
de Janeiro State University, where the mice were lodged together in our animal facilities. The
Committee for the Care and Use of Laboratory Animals of the Rio de Janeiro State University,
approved the protocols of the experiments described. Eight week old mice of TR and TS
strains were exposed to 50 transcutaneous cercariae obtained from Biomphalaria glabrata,
infected with S. mansoni (BH strain) maintained in the Reference Laboratory in Malacology of
Oswaldo Cruz Institute, Fiocruz, RJ, Brazil. Adult worms, acquired by perfusion of TS and TR
mice 8 weeks after infection, were collected from the portal and mesenteric veins in PBS pH
7.2 during necropsies of the abdominal cavity. Five mice of each sex were used. The helminths
were fixed for 2 h at room temperature or overnight at 4 ºC in 2.5% glutaraldehyde, with 4%
freshly prepared paraformaldehyde and 5mM calcium chloride in 0.1 M sodium cacodylate
buffer, pH 7.2. For SEM, the fixed adults were washed with cacodylate buffer, postfixed in 1%
OsO4 in 0.1 M cacodylate buffer containing 0.8% potassium ferricyanide and 5 mM CaCl2,
washed with cacodylate buffer, dehydrated in ethanol, processed in a criticalpoint drier
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with CO2, sputter-coated with gold, and examined in a Zeiss DSM 962 scanning electron
microscope operating at 15 kV. For TEM, fixed adults were postfixed as described for SEM and
dehydrated in acetone series and embedded in Spurr’s resin. Thin sections were collected on
copper grids, counterstained with uranyl acetate and lead citrate and then observed with
a JEOL JEM-1011 Electron Microscope. Results: SEM observations confirmed our previous
results which male worms recovered from TS mice had a low density of tubercles throughout
the tegument. In these specimens, the tubercles of the median portion of the body were
dispersed and intumesced, the distance between tubercles varying from 16.30 to 68.48 µm,
and the tubercles presented fewer spines than those in worms from TR mice. TEM results
confirmed and emended our findings by SEM. In the observations by TEM the parasites
recovered from TS mice presented tubercle swelling with blunted and immersed spines,
increased sensory organelle numbers, fusion and tegumental ridge peeling. In female worm
we observed lysis of interstitial tissues distributed among the degenerated vitelline cells. In
vitelline cells, the granular endoplasmic reticulum disappeared, the vitelline balls were fused
together or collapsed and vacuoles were observed with residual bodies. Conclusions: The
alterations observed by SEM in the tegument of parasites recovered from TS mice and those
revealed by TEM, in particular in the vitelline cells, suggest consequences on reproductive
success of the parasite. This reinforces the hypothesis that such alterations reflect the
interaction of the parasite with the immune regulatory environment of TS mice.
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CPg-Odn IS an aLtERnatIvE adJUvant tO BE USEd In a vaCCInE fORMULatIOn
agaInSt SCHIStOSOMIaSIS
Juliano Michel de araújo; tatiane teixeira de Melo; Ludmila zanandreis de Mendonça; Paulo
Marcos zech Coelho; Cristina toscano fonseca;
Centro de Pesquisa René Rachou, Fiocruz, MG, Brasil
Introduction: Schistosomiasis is the most important human helminth infection in terms of
global mortality. This parasitic disease affects more than 200 million people worldwide causing
more than 250,000 deaths per year. Recently, King et al have associated schistosomiasis with
anemia, pain, diarrhea, exercise intolerance, and under-nutrition that results from chronic
infection. Current schistosomiasis control strategies are mainly based on chemotherapy but,
in spite of decades of mass treatment, the number of infected people remains constant.
A vaccine that induces even a partial reduction in worm burdens could considerably
reduce pathology and limits parasite transmission. The larval schistosomula are the most
susceptible parasite life stage that interacts with host immune system and the interface of
this interaction lies primarily on the parasite tegument. Schistosoma is responsible for many
biological functions, including nutrient uptake, immune evasion and attachment. Several
studies in the past have focused on the identification of Schistosoma surface antigens related
to vaccine development and new advances in proteomics have allowed the characterization
of such tegument antigens, and some of them have shown to be highly protective in the
mouse model. Our group demonstrated that Smteg (schistosomula tegument) is able to
activate DC inducing an up-regulation of CD40 and CD86 and production of IL-12 and TNF-α.
In another experiment, mice immunization with smteg/freund‘s adjuvant was able to induce
partial reduction on worm burden (43-48%), on eggs trapped in the host tissue, and also in
eggs eliminated in the faeces (60%) besides inducing a Th1 response. Therefore, the purpose
of this study is to evaluate the effect caused by immunization with Smteg in the presence
of aluminum hydroxide and/or CpG-ODN. Methodology: Six to eight weeks-old female mice
C57BL/6 and cercariae of S. mansoni (LE strain) were obtained from Rene Rachou Research
Center (Fiocruz, Brazil). Cercariae of Schistosoma mansoni were mechanically transformed
into schistosomula and the tegument was removed with 0,3M of CaCl2 as previously
described (Durães et al 2003). C57BL/6 mice were divided into five groups of ten mice each.
Mice were subcutaneously injected with 25µg Smteg on days 0, 15 and 30. The Smteg or PBS
was formulated with Alum/CpG-ODN or Alum alone. Thirty days after the third immunization
mice were challenged with 100 cercariae and fifty days after challenged mice were perfused.
The measurement of specific anti- Smteg IgG, IgG1 and IgG2c antibodies was performed by
ELISA. Two different methods were performed to determine the number of eggs in the fecal
material: HPJ (Hoffman, W.A.; Pons, J. A.; Janner, S. L, 1934) and miracidia hatching (Juberg
et al. 2008). The statistical analyses were performed with Student’s t-test using the software
package GraphPad Prism 4.0. Results: Protective immunity induced by Smteg vaccination was
evaluated 50 days after challenge infection. It was observed that immunization with SmTeg/
Alum/CpG induced a 40% reduction on female worms, 46% on male worms and 43% in total
worm compared to the group inoculated with PBS/alum/CpG. No statistically significant
reduction on worm burden was observed between mice immunized with Smteg/Alum and
PBS/Alum. Regarding the number of eggs/miracidia eliminated with the faeces we observed
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a 56% reduction in the number of eggs/gram of faeces between the group immunized with
SmTeg/Alum/CpG and the group inoculated with PBS/Alum/CpG, and 35% reduction in the
miracidia hatching. Among the groups Smteg/Alum and PBS/Alum the reduction on the
number of eggs or miracidia/gram of faeces were respectively 44% and 35. A reduction of 28%
in the number of eggs trapped in the liver and of 24% in the number of eggs trapped in the
intestine was observed in Smteg/Alum immunized group compared to PBS/Alum group and a
33%, 36% and 35% reduction on eggs trapped in the liver, intestine and total respectively was
observed between Smteg/Alum/CpG and PBS/Alum/CpG. We did not observe a reduction
on female fecundity. Statistically significant levels of IgG, IgG1 and IgG2c were observed in
the group vaccinated with SmTeg/Alum/CpG compared to the group inoculated with PBS/
Alum/CpG. In the group of mice immunized with Smteg/Alum significant levels of IgG and
IgG1 were observed after second immunization compared to PBS/Alum group. Conclusions:
We conclude that Alum/CpG represent an alternative adjuvant formulation to be used in
association with Smteg, since this formulation was able to induce similar levels of protection
as the levels induced by Smteg/Freund‘s formulation.
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EvaLUatIOn Of tHE n-aCEtYLCYStEInE In tHE IMMUnOPatHOLOgY Of tHE
ExPERIMEntaL SCHIStOSOMIaSIS ManSOnI
andré de Lima aires; Renata alexandre Ramos Silva; tiago Moreira alves feitosa; Luiz
Henrique de Souza teixeira; giuliana viegas Schirato; nicodemos teles de Pontes filho; Sidcley
Bernardino de araújo; valdênia Maria Oliveira Souza; vlaudia Maria assis Costa; Elizabeth
Malagueño de Santana; Mônica Camelo Pessoa de azevedo albuquerque
Universidade Federal de Pernambuco, PE - Brasil
Introduction: The main pathogenic element of the Schistosoma mansoni infection are the
worms eggs that are retained in the intestinal mucosa or are carried to liver which is the
primary site of pathological alterations. In these organs, antigens released by miracidium
stimulate cells and host cytokines favoring the granulomatous inflammatory reaction that
results in a fibrous scar. Praziquantel (PZQ) is an efficient drug against S. mansoni but does
not change the histopathological lesions already installed that may result in irreversible
consequences. In humans and experimental animals, N-acetylcysteine (NAC) has attenuated
damage in chronic liver disease due to its anti-inflammatory, antioxidant and hepatic
detoxifying properties. Clinically, it is employed for treatment of a variety of diseases including
cystic fibrosis, biliary cirrhosis and toxicity associated to liver lesions. NAC contributes to the
synthesis of glutathione, one of the natural detoxifying agents most important on liver. This
drug directly acts on reducing levels of superoxide radicals diminishing the oxidative stress as
well as inhibits the releasing of tumor necrosis factor-α, the activation of pro-inflammatory
cytokines and apoptosis. Present work evaluated the actions of N-acetylcysteine and/
or praziquantel in immunopathologic modulating in the different phases of experimental
infection by Schistosoma mansoni. Methodology: Swiss Webster female mice were
percutaneously infected by 50 S. mansoni cercariae. Three experimental groups corresponded
to the acute (G1), intermediary (G2), and chronic (G3) phases of infection. Each group was
divided in 4 subgroups (n = 9) according to the following therapy: NAC, PZQ, NAC + PZQ and
control group that only received the drug vehicle. Administration of NAC, at 200 mg/Kg/
day, orally, in the acute phase of infection began one day after infection and was finalized
after 59 consecutive days. In the intermediary and chronic phases, NAC was administered
at the same dosage from the 45th day after infection and was maintained during 45 and
75 days respectively. Orally administration of 100 mg/Kg/day of PZQ occurred from 45th
to the 49th day after infection. Mice were sacrificed 60, 90, and 120 days post-infection
corresponding to the acute, intermediary, and chronic phases of infection respectively. At the
end of treatment, the histopathological and morphometrical analysis of liver tissue and the
determination of levels of interleukin-4 (IL-4), interleukin-10 (IL-10), interferon-ɣ (IFN-ɣ), and
nitric oxide (NO) in splenocyte culture were performed. Results: According to the employed
therapy, microscopical analysis revealed schistosomal granulomas of variable cellular
composition and organization with respect to the time of infection. In the different phases of
infection, the average number of liver granulomas in controls and NAC subgroup presented
significant difference (p < 0.001) as compared to the PZQ and NAC + PZQ subgroups with
percentage reduction of 71.09% and 94.07% respectively. Considering the mean diameter
of granulomas, in the acute phase (G1) there was a slightly reduction between G1-NAC
(312 µm ± 14.914) and G1-PZQ (321.1 µm ± 12.64) as compared to G1-control (340.1 µm
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± 7.73). However, intragroup analysis of the intermediary (G2) and chronic (G3) phases
showed significant reduction (p < 0.001) as compared to control subgroup. This reduction
varied between 35.68 (G2-NAC) and 44.49% (G2-NAC + PZQ). Intergroup analysis revealed a
natural modulation of granulomas diameter during infection, but NAC and/or PZQ increased
this modulation significantly. In the acute phase, the fibrosis degree on liver tissue for all
subgroups was classified as mild. However, in the intermediary and chronic phases, 77.7%
of animals treated with NAC and NAC + PZQ presented mild degree of fibrosis while control
and PZQ subgroups showed a fibrosis grade ranging from intermediary to intense. At the 60
days of infection, levels of IL-10 were well reduced (p < 0.001) in all subgroups, but at 90 days
it was observed a significant increasing (p < 0.001) in NAC and NAC + PZQ subgroups and at
120 days the IL-10 levels were reduced in these last subgroups. It was evidenced a negative
modulation of NAC in the IFN-ɣ levels and NO in the NAC and NAC + PZQ subgroups in all
phases of infection. Additionally, IL-4 levels in NAC subgroups were very similar to controls.
Conclusions: According to our results, use of NAC or NAC + PZQ may reduce morbidity in mice
infected by S. mansoni once we observed a significant reduction in size of granulomas and
lower collagen deposition. Furthermore, treatment with NAC reduced levels of IFN-ɣ and
NO and it increased the levels of IL-10. Finally, in this experimental model, NAC seems to
attenuate damage in liver tissue of schistosomotic mice.
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tHE EffECt Of SCHISTOSOMA MAnSOnI antIgEnS In dOWn-MOdULatE tHE
InfLaMMatORY RESPOnSE In tH2-MEdIatEd dISEaSE
Luciana Santos Cardoso; Sérgio Costa Oliveira; alfredo Miranda góes; Ricardo Riccio Oliveira;
Robson da Paixão de Souza; Edgar M Carvalho; Maria Ilma araujo
Serviço de Imunologia, HUPES/UFBA, BA - Brasil
Introduction: Schistosoma mansoni infections or their products can protect against allergic
disease since they suppress the host immune response. The mechanisms underlying this
association may include the production of regulatory cells and cytokines. The objective
of this study was to evaluate the immune response induced by the S. mansoni antigens
Sm22.6, PIII and Sm29 in cells of asthmatic individuals, and the ability of these antigens
in induce IL-10 production and suppress the production of IL-5. Methodology: Peripheral
blood mononuclear cells (PBMC) of asthmatics free of helminth infections were stimulated
in vitro with the S. mansoni antigens Sm22.6, PIII and Sm29 in the presence or absence of
the antigen 1 of the Dermatophagoides pteronyssinus (Der p1). Cytokines were measured in
the supernatants of PBMC cultures using a sandwich ELISA. The IL-10 producing cells were
assessed by flow cytometry. Results: It was observed a high production of IL-10 by cells of
uninfected asthmatic individuals in response to the S. mansoni antigens (384 ± 287, 492 ±
281, 473 ± 278 pg/ml to Sm22.6, PIII and Sm29, respectively). The Th1-signature cytokines
IFN-ɣ was detected at mean levels around 100 pg/ml to all tested antigens, being higher in
cultures stimulated with PHA (1549 ± 341 pg/ml). The Th2 cytokines IL-5 and IL-13 were
also measured and the levels of IL-5 were under the limit of detection to the three tested S.
mansoni antigens (15.6 pg/ml), being high only in cultures stimulated with PHA (763 ± 247
pg/ml). The mean levels of IL-13 were bellow 100 pg/ml, with the exception to the cultures
stimulated with PHA (835 ± 668 pg/ml). We also investigated the phenotype of IL-10 producing
cells in cultures stimulated with the S. mansoni antigens and the main source of this cytokine
were CD4+CD25+ and CD14+ cells. Moreover, we added the different S. mansoni antigens
to the cultures stimulated with Der p1 and it resulted in an increased levels of IL-10 (Der
p1= 209±130pg/mL; Der p1+Sm22.6=1190±595pg/mL, p<0.0001; Derp1+PIII=896±466pg/
mL, p<0.0001; Derp1+Sm29=653±289pg/mL, p<0.0003), and reduced levels of IL-5 (Der p1=
296±231pg/mL; Der p1+Sm22.6=94±154pg/mL, p=0.02; Derp1+PIII=143±197pg/mL, p=0.03;
Derp1+Sm29=96±86pg/mL, p<0.01). Conclusions: The S. mansoni antigens evaluated in this
study induced the production of the regulatory cytokine IL-10 and down-modulated the
Th2 immune response which participates in the pathology of asthma. These antigens could
represent new strategies to prevent allergic diseases. Financial support: CNPq.
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IMUnOdOMInanCE Of tHE SM-P40 antIgEn COntRIBUtES tO tHE dEvELOPMEnt Of
SEvERE SCHIStOSOMIaSIS ManSOnI
Eduardo finger, thaissa Melo galante Coimbra, daniel Maurício dos Santos
Santa Casa de São Paulo, SP, Brasil
Introduction: In experimental murine schistosomiasis, the H-2k CBA and C3H (high pathology)
strains differ from the H-2b C57BL/6 and H-2d BALB/c (low pathology) strains for developing
a much severer CD4 T cell mediated liver immunopathology and a high Th1/Th17 type
CD4 T cell response against the schistosome major egg antigen Sm-p40 and in particular,
its immunodo-minant epitope: peptide 234-246. Low pathology strains do not react
significantly against this antigen. This marked correlation between strong Th1/Th17 antiSm-p40234-246 reactivity and severe liver immunopathology suggests that the former may
polarize the anti-egg immune re-sponse and lead to the latter. A possible explanation for how
a single epitope may determine the course of the entire anti-egg immune response could be
the presence of restrictive immunodominance, a phenome-non induced when vast epitope
availability combines with a very high affinity for a selective major histocompatibility class
II haplotype (MHCII), to allow that epitope to dominate MHCII presentation on the surface
of antigen presenting cells. Restrictive immunodominance strongly favors the development
of a vigorous Th1 type response against that dominant epitope. With Sm-p40 comprising at
least 10% of the whole egg protein content and Sm-p40234-246 representing the highest
affinity epitope for the unusually stringent H-2k haplotype, high pathology strains present
conditions perfect for the establishment of restrictive immunodominance of Sm-p40234-246
so hypothetically, this situation might generate such a strong Th1/Th17 response against it
that not only it polarizes the anti-egg immune response, but it also thwarts all the attempts
at modulation and the result is severe liver immunopathology. In order to test the hypothesis
that restrictive immunodominance of Sm-p40 contributes to the development of severe liver
immunopathology we: a) analyzed the liver immunopathology of non-H-2k infected mice
whose MHCII restriction pattern is likely to produce restrictive immunodominance of Sm-p40
and b) analyzed the liver immunopathology of infected CBA mice immunized with a cocktail
of epitopes designed to neutralize Sm-p40234-246 immunodominance. Methodology: NonH-2k strains likely to develop restrictive immunodominance of Sm-p40 were identi-fied by
analyzing the restriction profile of Sm-p40 in several MHCII haplotypes with the RANK-PEP
algorithm. The strain SJL was selected for being restricted to present a single epitope of Smp40 (CBA presents 2). For analysis of SJL liver immunopathology, mice were infected intraperitoneally with 150 S. mansoni cercariae, euthanized 7 weeks-post-infection(wpi) and
their livers were processed and submitted to morphometric analysis of liver granulomas.
For the analysis of the neutralization of Sm-p40234-246 restrictive immunodominance, 3
groups with 3 CBA mice each were infected intraperitoneally with 150 S. mansoni cercariae.
Group 1 was sensitized with complete Freund’s adjuvant (CFA) at 2 and 4 wpi. Group 2 was
sensitized on the same dates with an emulsion of CFA and a cocktail of 4 synthetic epitopes
from egg proteins, selected for their ability to compete for I-Ak presentation with an equal
or higher affinity than Sm-p40234-246, and group 3 was left as control. All groups were
euthanized at 7 wpi and their livers underwent morphometric analysis of liver granulomas.
Results: Morphometric analysis of the immunopathology of SJL mice revealed that this
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strain develops as severe an immunopathology as the CBA strain. In fact, the average liver
granuloma in SJL mice was larger than those measured on the CBA strain (176.101±12.508
um2 for the SJL versus 144.693±21.890 um2 for the CBA and 106.439±8.205 um2 for the
C57BL/6)*. Analysis of the liver immunopathology of the 3 groups of CBA mice revealed that
treatment with the cocktail emulsion not only prevented a CFA mediated aggravation of liver
immunopathology but it also significantly reduced the severity of liver immunopathology
(173.873±37.522 um2 for the untreated CBA control versus 233.650±41.302 um2 for the
CBA immunized with CFA alone for the CBA and 90.256±15.006 um2 for the CBA immunized
with CFA/cocktail). * mean±SEM Conclusions: These results provide evidence that restrictive
immunodominance might be a pathogen-ic force in the establishment and severity of CD4 T
cell mediated diseases and that its neutralization might be an effective therapeutic strategy
that deserves further exploration.
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RECOgnItIOn Of SCHIStOSOME tEgUMEnt PROtEInS BY SERa fROM PUtatIvE
RESIStant and CHROnICaLLY InfECtEd HUManS USIng a PROtEIn MICROaRRaY
Soraya gaze; Patrick driguez; angela trieu; Jeffrey Bethony; fernanda Cardoso; Rodrigo
Correa-Oliveira; Phil felgner; donald McManus; denise doolan; alex Loukas
James Cook University, Australia
Introduction: Schistosomiasis is a neglected tropical disease that affects more than 207 million
people worldwide, with an estimated 700 million people at risk in 74 endemic countries.
Praziquantel (PZQ) is the only available treatment against all forms of schistosomiasis, and
long term control of the disease with PZQ alone is unrealistic. Immunoepidemiological
investigations have shown that a very small percentage of exposed populations in Brazil are
naturally resistant to schistosomiasis despite never being treated with PZQ. We propose
to evaluate the humoral response of putative resistant and chronically infected individuals
to schistosome tegument antigens spotted onto a protein microarray to understand the
protective mechanisms involved and the antigens targeted by these individuals. Methodology:
To identify S. mansoni antigens that are uniquely/preferentially recognized by resistant
compared with susceptible individuals, we constructed a protein microarray consisting of
60 S. mansoni (Sm) and 180 S. japonicum (Sj) proteins, predominantly those found in the
tegument. We screened the array with sera from individuals from Minas Gerais, Brasil, who
were putatively resistant (PR- n=15) to schistosomiasis mansoni, chronically infected (CIn=15) or unexposed to the parasite (n=7). We evaluated IgG subclass and IgE responses to
the proteins in the array and compared their recognition profiles. One-way ANOVA followed
by Bonferoni test were used to evaluate significance between the groups (p<0.05). Results:
PR and CI groups recognised many of the proteins on the array. There was extensive crossreactivity between S. mansoni and S. japonicum antigens. In general, sera from the PR yielded
higher IgG1, IgG3 and IgG4 levels than those from the CI group compared to unexposed
controls. One tegument membrane protein, Sm200, was preferentially recognised by IgG3
but not IgG4 (marker of chronicity) fromthe PR compared to the CI group, indicating that this
protein is a good candidate vaccine antigen. One unknown protein and Sm29 had higher IgG4
responses in the PR compared to the CI group. Thirty-nine (39) of 60 S. mansoni proteins had
higher IgE levels in the PR compared with the CI group, strongly implicating IgE responses
as a protective mechanism. Conclusions: PR individuals had elevated IgE responses to S.
mansoni proteins compared to CI individuals, implying a pivotal role in resistance. However,
the presence of a pre-existing IgE response to a vaccine antigen can promote an allergic
response at vaccination, precluding the development of such antigens as components of
recombinant helminth vaccines for infected individuals. This is the first report describing the
construction and screening of a schistosome protein array with human sera, and the results
presented will be valuable in the up- and down-selection of antigens for development of a
subunit vaccine for human schistosomiasis.
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tHE USE Of SCHISTOSOMA MAnSOnI antIgEnS tO PREvEnt tHE InfLaMMatORY
RESPOnSE In vItRO In CUtanEOUS LEISHManIaSIS.
aline Michelle Barbosa Bafica; giuseppe tittone varella; Ricardo Riccio Oliveira; Luciana
Santos Cardoso; alfredo góes; Sérgio Costa Oliveira; alex Loukas; Edgar Marcelino Carvalho;
Maria Ilma araújo
Serviço de Imunologia-HUPES/UFBA, BA, Brasil
Introduction: Leishmaniasis is one of the most important infectious diseases worldwide.
The expression of the disease depends on the host immune system, parasite virulence
and environmental factors. The control of Leishmania infection depends on the immune
response mediated by T cells (Th1). They produce cytokines, especially IFN-ɣ that are able
to activate tissue macrophages and support parasite restraint. However, when this immune
response is deregulated and there is high production of IFN-ɣ and TNF intense tissue damage
occurs, leading to development of the mucosal and cutaneous form of leishmaniasis. Recent
experimental studies have shown that S. mansoni infection or parasite products by induce
regulators cells and cytokines are able to down-modulate the Th1 inflammatory response
involved in some autoimmune diseases, such as type-I diabetes, arthritis and psoriasis.
Our hypothesis is that S. mansoni antigens rSm29, rTSP2 and PIII are able to down regulate
the inflammatory response of LC patients. Objective to evaluate the effects of S. mansoni
antigens on the immune response induced by the soluble Leishmania antigen (SLA) in
cutaneous leishmaniasis patients, we examined the production of cytokine important in
the development of an immune response, as well as intracellular cytokine expression by
monocyte from CL patients. Methodology: The CL patients included in this study live in Corte
de Pedra, an endemic area of Leishmania Bahia. The proinflammatory cytokines IFN-ɣ and
TNF and the regulatory cytokine IL-10 were measures in the supernatants of peripheral
blood mononuclear cell (PBMC) culture stimulated with SLA in the presence and absence
of rSm29, rTSP2 and PIII, using a sandwich ELISA technique. Additionaly, PBMC cells were
stained with fluorochrome conjugated antibodies for the expression of surface molecules
CD14+ and CD16+, intracellular cytokines (IL-12, TNF and IL-10). Results: The addition of
rSm29 to the cultures reduced the levels of IFN-ɣ in 7 of 13 patients (53.8%; mean reduction
48.8%). The PIII antigen was also able to decrease the levels of IFN-ɣ in 7 of 12 individuals
(58.3%; mean reduction 55.7%). The three antigens used in the study were able to down
modulate the production of TNF in 30% (mean reduction variation 35-43%). On other hand,
the addition of Sm29 and PIII resulted in an increase in IL-10 production in 100% and 70%
of the individuals, respectively (mean increase 808.7 and 101.5 %). We also evaluated the
frequency of classical and proinflammatory monocytes expressing inflammatory cytokines
after in vitro stimulating with SLA and S. mansoni antigens. Regarding proinflammatory
monocyte, the addition of rSm29 resulted in decrease of the CD14+CD16+ cells expressing
IL-12 in 8 of 14 patients (57.1; mean reduction 60.8%) and TNF in 5 of 12 patients (41.6;
mean reduction 72%). The rTSP2 antigen was able to reduce CD14+CD16+ cells expressing IL12 in 5 of 10 LC patients (50%; mean reduction 67.2%). There was no significant variation in
the frequency of CD14+CD16+ cells expressing IL-10 by the presence of S. mansoni antigens.
In cultures stimulated with rSm29 there was decrease in CD14+ cells expressing IL-12 in 6
of 14 patients (42.9%; mean reduction 49.9% ) and TNF in 6 of 13 patients (46.2%; mean
126
reduction 49.9%). In the presence of PIII there were decreases in CD14+ cells expressing IL-12
in 9 of 14 patients (64.3%; mean reduction 68.9%) and TNF in 5 of 13 patients (38.5%; mean
reduction 55.1%). The three antigens used in the study were able to increase the CD14+
cells expressing IL-10 (mean increase variation 125-225%). Conclusions: The S. mansoni
antigens used in this study particularly rSm29 and PIII down-modulated the production of
IFN-ɣ and and TNF and up-regulated IL-10 production in cells of CL patients in response to
the Leishmania SLA antigen in vitro. Moreover the antigens rSm29, TSP2 and PIII were able to
reduce the frequency of classical and proinflammatory monocytes expressing IL-12 and TNF.
These antigens therefore have the potential to be used as modulators of the inflammatory
response in leishmaniasis. FINANCIAL SUPPORT - CNPQ (Universal 479417/2008 begin of the
skype highlighting 479417/2008 end of the skype highlighting).
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dIffEREntIaL PROtEIn PaRtItIOnIng dURIng tEgUMEnt ISOLatIOn Of tHE
PaRaSItE SCHISTOSOMA MAnSOnI
Priscila tavares Meneses; Sonaly Cristine Leal; tiago ferreira Leal; Milton Hercules guerra de
andrade; R. alan Wilson; William de Castro Borges
Universidade Federal de Ouro Preto, MG, Brasil
Introduction: Representing the important host-parasite interface in the context of
schistosomiasis, the worm tegument has been the subject of a variety of protocols for its
isolation. These included salt, detergent and freeze-thaw extraction techniques. Although
markers of surface molecules such as alkaline phosphatase have been traced, to assess
enrichment of tegument proteins in such preparations, the aforementioned protocols do
not provide a complete recovery of tegument components, especially those located at
the membranocalyx and plasma membrane. As a consequence, most of the proteomic
investigations of the S. mansoni tegument do not allow for accurate conclusions regarding
protein abundance and, more importantly, insights into protein location at the parasite
surface remain desirable in the search of the ideal vaccine target against the disease.
Methodology: As a first step, our study proposed a deviation of the freeze-thaw isolation
protocol developed in the 80´s, with the exclusion of the sucrose gradient employed for
purification of the membranes. Instead, we established that all proteins extracted during
the freeze-thaw technique would be recovered by ultra-centrifugation through which five
fractions termed S1S, S1P, S2P, S2SS and S2SP were generated. Our next strategy involved
the investigation of protein partitioning using 1D Western blotting technique. In total, ten
antibodies targeting S. mansoni proteins were used to probe PVDF membranes containing
equal amounts of the obtained fractions. Among them, selected soluble/cytoskeletal
components such as Sm22.6, and membrane-bound molecules such as nutrient transporters,
tetraspanin 2 and annexins were investigated alongside host molecules detected using an
anti- red blood cell antibody. Results: Firstly, our results confirmed the presence of tegument
proteins throughout fractions, e.g S1P and S2SP, not previously considered in the original
freeze-thaw isolation protocol. Then, our results revealed nutrient transporters, tetraspanin
2, annexin 2, CD59, Sm29, carbonic anhydrase and Sm200 as genuine constituents of the
tegument membranes. A second class of molecules represented by annexin 1 and Sm22.6
were found distributed in both soluble and membrane fractions. The only exclusively soluble
protein detected was SmKK7, found primarily in the S1S fraction, which is in agreement
with its proposed location at nerve terminal endings. Conclusions: We concluded that
the additional tegument fractions derived from this alternative isolation protocol allows
for a global analysis of protein composition at the worm surface. At present, proteomic
investigations of these membrane fractions are under way with the potential to orient and
direct selection of novel vaccine candidates.
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tHE SCHISTOSOMA MAnSOnI PHYLOME: EvOLUtIOnaRY HIStORIES tO IMPROvE
fUnCtIOnaL PREdICtIOnS and gEt InSIgHtS IntO PaRaSItE BIOLOgY
Larissa Lopes Silva; Marina Marcet-Houben; Laila alves nahum; adhemar zerlotini neto; toni
gabaldón; guilherme Oliveira
Introduction: Schistosoma mansoni (Platyhelminthes: Trematode) is one of the etiologic
agents of human schistosomiasis, a tropical neglected disease that is endemic in 76
countries where 210 million people are infected and other 779.000 live in areas of risk of
infection. The parasite predicted proteome was recently published and contains over 13.000
proteins. However, more than 40% remain without a predicted function or experimental
characterization. The goal of the present study was to improve the functional annotation of
the S. mansoni proteome and to get insights into parasite biology by applying a phylogenomic
approach. Methodology: Proteomes derived from 17 fully sequenced eukaryotic genomes
were downloaded from online databases, including one plant, three fungi, eight invertebrates,
one urochordate, one cephalochordate, and three vertebrates, totaling more than 340.000
proteins. The reconstruction of the evolutionary histories of all proteins encoded in the S.
mansoni genome was performed using an automatic pipeline as implemented in PhylomeDB
(http://phylomedb.org). Results: The S. mansoni phylome includes 8.818 phylogenetic trees
derived from the comparative analyses of 13.285 parasite proteins and their homologs in
16 other organisms. By using this approach, we could transfer functional annotations (Gene
Ontology terms) to 5.507 S. mansoni proteins, 946 of which were previously annotated as
“hypothetical” or “expressed protein”, corresponding to proteins with a completely unknown
function. Conclusions: Besides promoting a significant improvement in the functional
annotation of the S. mansoni proteome, this approach has provided useful information
about the parasite’s genome evolution such as the identification of gene duplication events.
Among the parasite paralog groups, we have identified tetraspanins, leishmanolysins, and
fucosyltransferases, that may be related to morphological or physiological specificities of
S. mansoni, which were previously proposed as drug targets. All sequence alignments,
phylogenetic trees, and functional annotation will be made publicly available through
PhylomeDB and SchistoDB (http://schistodb.net) databases providing a powerful resource
for the scientific community. Financial support: National Institutes of Health - NIH/Fogarty
International Center (Grants: D43TW007012 and 2D43TW007012), Secretaria de Ciência,
Tecnologia e Ensino Superior de Minas Gerais and Fundação de Amparo à Pesquisa do Estado
de Minas Gerais, SECTES/FAPEMIG, Brazil (Grant: 1181/08).
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tOWaRdS an UndERStandIng Of tHE EPIgEnEtICS Of SCHIStOSOMES
Céline Cosseau; Julie Lepesant; Jérome Boissier; Julien Portella; Cécile Perrin; Christoph
grunau
University of Perpignan - Tropical and Mediteranean Biology, France
Introduction: As probably all eukaryotes, Schistosomes use a supplementary information
transmitting system, the epigenetic inheritance system (EIS) to shape genetic information
and to produce different phenotypes. In contrast to other important parasites, the study
of epigenetic phenomena in Schistosomes is still in its infancy. Nevertheless, we begin
to get hold of what goes on behind the epigenetic scene in this parasite. We will resume
what our lab has contributed to the understanding of epigenetics in S. mansoni in the last 3
years. Methodology: We have developed techniques of chromatin immunoprecipitation and
associated the necessary bioinformatics tools that allow us to run genome-wide chromatin
studies on S. mansoni at different stages of the life cycle. Results: Using these techniques we
could show that chromatin structure changes during the life cycle of the parasite and that
difference in chromatin structure exist between different parasite strains. We proved that
direct influence on the EIS through treatment with inhibitors of histone-modifying enzymes
leads to developmental arrest and phenotypic changes. In addition, chromatin structure
changes are associated with transcription and silencing of repetitive satellite-style sequences.
We have shown that these sequences are specific to female and we will hypothesize on the
role of these repeats in the sexual dimorphism observed at the adult stages. Conclusions: In
summary, our data show that epigenetic phenomena are probably participating in many, if
not all pathways that require control of gene expression in S. mansoni and we anticipate that
these help to unravel the evolution and adaptation phenomena of the Schistosomes.
130
nUCLEaR ExPORt and SECREtIOn Of SCHISTOSOMA MAnSOnI HMgB1 PROtEIn IS
MEdIatEd BY PHOSPHORYLatIOn
Isabel Caetano de abreu da Silva; vitor Coutinho Carneiro; Renata de Moraes Maciel; Rodrigo
furtado Madeiro da Costa; daniel Rodrigues furtado; francisco Meireles; Mario alberto
Cardoso da Silva-neto; franklin david Rumjanek; Marcelo Rosado fantappié
Universidade Federal do Rio de Janeiro, RJ, Brasil
Introduction: The High Mobility Group Box (HMGB) family represents a class of evolutionarily
highly conserved and abundant chromosomal proteins that recognize and change the topology
of DNA. HMGB1 contains two copies of homologous DNA binding motifs named HMG-boxes A
and B. It also contains an acidic rich C-terminus. HMGB1 is both a nuclear factor, and a secreted
protein. In the nuclei, HMGB1 can facilitate the assembly of complexes involved in recombination
and transcription, as well as nucleosome remodeling. Outside the cell, mammalian HMGB1
binds with high affinity to RAGE (the receptor for advanced glycation end products), and to
multiple Toll-like receptors, acting as a potent mediator of inflammation. Previously, we cloned
the HMGB1 protein from the parasite Schistosoma mansoni (SmHMGB1), which showed high
similarity among their HMG box domains with its orthologues but display a shorter acidic tail,
comprising only 5 acidic residues. Post-translational modifications, mainly acetylation and
phosphorylation, of HMGB1 proteins regulate its DNA binding capacity as well as its cellular
localization. We have previously showed that acetylation of SmHMGB1 can translocate the
protein from the nucleus to the cytoplasm, following its secretion to the extracellular milieu. In
this work we investigated the role of phosphorylation in SmHMGB1 cellular traffic. Methodology:
In silico prediction of SmHMGB1 phosphorylation revealed putative sites for protein kinases.
In vitro phosphorylation assays were carried out using three different protein kinases: Casein
kinase 2 (CK2), Protein kinase A (PKA) and Protein kinase C (PKC). Heterologous transfection
system with SmHMGB1-EGFP fusion protein and the phosphatase inhibitor okadaic acid were
used to evaluate the role of phosphorylation in SmHMGB1 cellular traffic. A CK2 specific inhibitor
TBB was also used. Western blotting analysis was performed to detect and access SmHMGB1
phosphorylation status in adult worms. Immunohistochemistry analysis was carried out with
liver sections of infected mice to follow SmHMGB1 staining in the granuloma. Results: In silico
prediction of SmHMGB1 phosphorylation revealed putative sites for protein kinases. In vitro
phosphorylation assays were carried out using three different protein kinases: Casein kinase
2 (CK2), Protein kinase A (PKA) and Protein kinase C (PKC). Heterologous transfection system
with SmHMGB1-EGFP fusion protein and the phosphatase inhibitor okadaic acid were used
to evaluate the role of phosphorylation in SmHMGB1 cellular traffic. A CK2 specific inhibitor
TBB was also used. Western blotting analysis was performed to detect and access SmHMGB1
phosphorylation status in adult worms. Immunohistochemistry analysis was carried out with
liver sections of infected mice to follow SmHMGB1 staining in the granuloma. Conclusions:
SmHMGB1 was specifically phosphorylated in vitro by CK2, PKA and PKC. Phosphorylation
did not alter the DNA binding actions of SmHMGB1. Phosphorylation regulated the traffic of
SmHMGB1 from the nucleus to the cytoplasm, as well as its secretion to the extracellular milieu.
Our data suggest that SmHMGB1 secreted by the eggs of the parasites might act as a proinflammatory molecule and as such, it might contribute to the development of the granuloma.
131
nUCLEOCYtOPLaSMIC SHUttLIng Of SCHISTOSOMA MAnSOnI HMgB1 IS REgULatEd
BY aCEtYLatIOn tHat dIRECtS It tOWaRd SECREtIOn
vitor Coutinho Carneiro; Isabel Caetano de abreu da Silva; Claudia neto Paiva; Renata de
Moraes Maciel dos Santos; flavia Lamarão; Marcelo Pelajo Machado; Hélio dutra; Marcelo
torres Bozza; Marcelo Rosado fantappie
Universidade Federal do Rio de Janeiro, RJ - Brasil
Introduction: HMGB1 is a nonhistone DNA-binding protein that functions as a structural
cofactor critical for proper transcriptional regulation in somatic cells. It promotes DNA bending
and facilitates the binding of several regulatory protein complexes to DNA. Despite the nuclear
function HMGB1 can also be passively and actively released to extracellular medium signaling
tissue damage or acting as a cytokine, respectively. The actively released protein plays an
important role in the immune response activating macrophages, T and B cells, maturating
dendritic cells, increasing vascular permeability and adhesion molecules expression. A
significant number of inflammatory diseases have been associated with high expression of
HMGB1 and their blockage brings a better prognostic in sepsis, arthritis and some parasitic
diseases. Methodology: A heterologous transfection systems using EGFP constructs were
carried out to evaluate the cellular traffic of SmHMGB1. Fluorimetric analysis was performed
to detect the secretion of EGFP-SmHMGB1 in culture media. Immunehistochemistry of liver
sections of S. mansoni infected mice was performed to evaluate the staining of SmHMGB1
in the granuloma. I addition, murine macrophages were treated with SmHMGB1 to detect
TNF-α and IL-1β by ELISA. The migration capacity of murine eosinophils was investigated by
the presence of SmHMGB1 as a potential chemotaxis factor. Results: We previously showed
that S. mansoni HMGB1 (SmHMGB1) was acetylated in vitro. To determine the biological
role of acetylation of SmHMGB1, HeLa cells were transfected with SmHMGB1-GFP and its
cellular traffic was followed within the cells. By fluorescence microscopy we showed that
SmHMGB1 was mainly present in the nucleus. However, when cells were treated with
sodium butyrate (NaB, a histone deacetylase inhibitor), SmHMGB1 were readily translocated
to the cytoplasm, and secreted to the extracellular medium. Importantly, we demonstrated
that the acidic tail of SmHMGB1 plays an important role in the nuclear exit and secretion of
the protein. We observed that S. mansoni eggs secreted significant amounts of SmHMGB1
and that egg-secreted SmHMGB1 were localized throughout in hepatic schistosomotic
granuloma. We showed that SmHMGB1 was able to induce the release of high amounts
of TNF-α and IL-1β from murine macrophages, thus acting as a pro-inflammatory cytokine.
Importantly, SmHMGB1 also acted as a chemokine, being able to recruit schistosomotic
eosinophils. Conclusions: We showed that acetylation plays a major role in cellular trafficking
and secretion of SmHMGB1. Our data strongly suggested that secretion of SmHMGB1 by the
eggs might contribute to the inflammatory reaction that leads to granuloma formation, by
activation and/or recruitment of host cells.
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In SEaRCH Of a PHaRMaCOPHORIC MOdEL fOR SCHIStOSOMICIdaL BEnzOdIazEPInES
Jean Pierre Barros thibaut; Carla Maria Souza Menezes; gildardo Rivera; nathália Couto dias;
Marina amaral alves; Eliezer J. Barreiro; Lídia Moreira Lima; françois noel
Universidade Federal do Rio de Janeiro, RJ, Brasil
Introduction: Clonazepam (CLO) and 3-methylclonazepam (MeCLO) 1,4-benzodiazepine
compounds induce contraction of adult male Schistosoma mansoni in vitro and have a
proven schistosomicidal activity in humans, but their mechanism of action is unknown.
We previously characterized the presence of two benzodiazepine binding sites in S.
mansoni (Noël et al., Parasitology 134: 1003, 2007), we also showed that the intense
and immediate worm’s musculature contraction induced by CLO and MeCLO is not
mediated by the parasite benzodiazepine binding sites (Thibaut et al., Eur J Pharmacol
606: 9, 2009). Here we decided to investigate the structural requirements for the in vitro
schistosomicidal effect of schistosomicidal benzodiazepines. Methodology: Six new N1modified clonazepam analogues were synthesized using clonazepam as starting material,
exploring a chemoselective N-alkylation using KF/Al2O3 in acetone, at room temperature,
in the presence of the appropriate alkyl halides (Lima et al., J Het Chem 42: 2005). The N1modified CLO substituents were CH3 ; CH2CH=CH ; CH2CO2H ; CH2Ph ; CH2CO2Me and
CH2CO2Et. Afterwards, we tested their schistosomicidal activity in vitro using the screening
assay recommended by the WHO. In accordance with our Institutional Ethical Committee for
animal care (CEUA, Nr DFBCICB011) adult worms were surgically removed from portal veins
and mesenteric of infected mice and distributed in tissue culture dishes in DMEM. The worms
were kept at 37°C in an atmosphere of 5% CO2 in air and were observed daily for 5 days
under a Nikon Eclipse TC-100 inverted microscope. The measurement of worm “motility”
is scored on an arbitrary scale: 4 = hyperactivity; 3 = normal activity; 2 = low activity, i.e.
occasional movement of head and/or tail; 1 = minimal activity, i.e. only gut movements; 0 =
total absence of motility even at the gut extremities. Worms were defined as ‘dead’ if they
remained without any movement of body (scale 0) during 1 minute. The molecular modelling
study based on stereoelectronic features was performed with the Spartan’08 software, using
the chemical structures constructed using the MMFF94 force field software. The elected
compounds for this study were: R-(-)- and S-(+)-meclonazepam, clonazepam, diazepam,
flunitrazepam, our N1-modified clonazepam analogues and the meclonazepam analogues
reported by Mahajan et al. (Bioorg & Med Chem Lett 18: 2333, 2008). Results: The six N1modified clonazepam analogues were not schistosomicidal nor disturbed the worm motility
and morphology. Based on the biological data and the molecular modelling approach, we
propose that the schistosomicidal activity is strictly related to stereolectronic features since
the presence of an oxo (preferential) or thienylic carbon (C2), the lack of substitution of
the amide nitrogen atom (N1), the presence of the imine bond (4- and 5-positions) and
the presence of an electron-withdrawal group at C7 seem to be essential features for the
schistosomicidal activity. Conclusions: The findings reported herein may aid to design new
meclonazepam analogues with special attention to the lack of substitution of the secondary
amide function.
133
dLCS aS nEW vaCCInE CandIdatES agaInSt SCHIStOSOMIaSIS
Patricia Placoná diniz; Erika nakajima; Patricia aoki Miyasato; toshie Kawano; Elizabeth
angelica Leme Martins
Instituto Butantan, Secretaria de Saúde Município de São Paulo, SP, Brasil
Introduction: Schistosomiasis is one of the most important helmintic diseases in the world.
It is present in 76 countries, with more than 200 million people infected and it is estimated
that more than 600 million people live in endemic areas. In 2003 extensive data from the
transcriptome of Schistosoma mansoni was made available. The information o the encoded
proteins allowed the analysis of protein function and improved the search for vaccine
candidates. The analysis of the transcriptome allowed the identification of three families of
proteins homologs to the mammalian dynein light chain (DLC). One of these was the L8 family,
with at least 18 members, all proteins with around 10 kDa. These proteins were found to be
expressed in the different stages of the S. mansoni life cicle. Two DLCs were recognized in
the tegument of S. japonicum, suggesting that they are exposed to the host immune system.
Considering these aspects we selected two DLCs from L8 family to be tested as vaccine
candidates. Methodology: To investigate the immunogenicity and antigenicity of two DLCs
from S. mansoni, to evaluate the protective profile in challenge assays with cercarias and to
analyse the development of hepatic granulomas that are involved in the main pathology of
Schistosomiasis we selected two genes of DLCs to be tested. They were cloned in E. coli for
protein expression, the purification was made by metal affinity cromatography. The purified
proteins were used for immunization assays. The sera generated against DLCs were tested
in ELISAs and Western Blot assays. After three immunizations with 10μg of purified protein
plus 0.3% allydrogel, mice were challenged with cercarias. After 45 or 55 days mice were
perfused and the worm burden and hepatic granuloma formation were analysed. Results:
Both DLCs showed to be very immunogenic increasing the IgG titers in the sera. Besides, the
group immunized with DLC1 increased the IgE levels before infection. After infection both
DLCs showed lower IgE levels when compared to the control groups. The DLCs showed to be
antigenic, since they were recognized by the sera of the control infectd mice. In the challenge
assays the DLCs showed a significant decrease in the worm burden (among 40 and 60%).
The granuloma analysis with 45 days after infection showed that the groups immunized
with DLCs had significative smaller ganuloma areas, up 70%, when compared to the control
group. After 55 days the granulomas from groups immunized with purified proteins were still
smaller (among 25 and 35%). Conclusions: The most promissing antigens tested as vaccine
candidates against Schistosomiasis shows a protective immune profile with 30-40% in the
decreasing of worm burden when allydrogel is used as adjuvant. Taking together, the results
of decreasing the worm burden and the granuloma size after immunization with purified
DLCs, they suggest that these proteins could be considered as very interesting vaccine
candidates, affecting the main causes of the pathology of Schistosomiasis, and they could be
included in a vaccine formulation against the disease.
134
EvaLUatIOn Of tHE IntERaCtIOn BEtWEEn PRIMaRY and SECOndaRY SPOROCYStS
Of SCHISTOSOMA MAnSOnI ExPOSEd tO tHE IntERnaL dEfEnSE SYStEM Of
BIOMPHAlARIA TEnAGOPHIlA, RESIStant and SUSCEPtIBLE tO tHE PaRaSItE
ana Carolina alves de Mattos; Raquel Lopes Martins-Souza; Paulo Marcos zech Coelho
Centro de Pesquisa René Rachou/Fiocruz, MG, Brasil
Introduction: Planorbidae of the genus Biomphalaria are the intermediate hosts of
Schistosoma mansoni. A restrict number of Biomphalaria species or lineages are susceptible
to a specific strain of S. mansoni. Biomphalaria has an internal defense system (IDS) composed
of cells (hemocytes) and soluble hemolymph factors that are stimulated in the presence of
parasites. In contrast to the host response, there are scape mechanisms of the parasite that
may assure surveiving and adaptability of the parasite to the host. One of these mechanisms
is mimetism or molecular masking, which would be the innate presence of molecules similar
to those of the host on the membrane surface of sporocysts, or the ability of the parasite to
absorb host antigens, respectively. These mechanisms may preclude the recognition of the
parasite by IDS of the host. The aim of this study was to evaluate the mode of action of IDS of B.
tenagophila Taim (resistant) and of Cabo Frio (susceptible) lineages in the presence of primary
sporocysts transformed in vitro and secondary sporocysts, derived from infected snails, by
means of cellular adhesion trials, viability and using fluorescent markers. Methodology: In
the trials carried out for evaluation of the cellular adhesion index and viability of primary and
secondary sporocysts, 20 parasites were exposed to the IDS fractions of B. tenagophila Taim
and Cabo Frio: total hemolymph (TH), fraction of hemocytes only (H), or only soluble fraction
(S). After 2h, a total of 100 sporocysts for each lineage was analyzed for obtention of cellular
adhesion index (CAI), using the protocol described by Castillo & Yoshino (2002). An arbitrary
value was attributed from 1 to 4, being CAI 1: without adhered hemocytes on the sporocyst
surface; CAI 2: up to 10 adhred hemocytes; CAI 3: between 11 and 50 hemocytes, and CAI
4: more than 50 adhered hemocytes. The viability of the sporocysts was assessed 6h postincubation, by means of 4% Trypan blue staining. To evaluate the presence of tegumental
damage, the parasites were submitted to the same conditions before described. Five hours
post-incubation they were marked with the fluorescent probe Hoechst 33258 (specific for
DNA), Glycine max (soybean) lectin (SBA), (specific for N-acethylgalactosamine) and AlexaFluor 488 faloidine (specific for actine filaments) for 30 min. Afterwards, the parasites were
washed and analyzed in a fluorescence microscope. Results: The hemocytes of both lineages
were able to adhere to the surface of primary and secondary sporocysts. When the mollusc
lineages were compared, it was verified that CAI in secondary sporocysts exposed to the
fraction H from Taim lineage was significantly higher (CAI = 2.277±0.01) than CAI detected in
secondary sporocysts exposed to the same fraction from Cabo Frio lineage (1.993±0.04) (p =
0.01). In the experiments related to viability, using primary sporocysts, when the proportion
of dead primary sporocysts after exposition to the fraction of the different lineages were
compared, statistically significant differences (p=0.02) were detected; 46.25% of the primary
sporocysts exposed to the TH fraction from Taim were found dead, whereas only 22.5% died
in the presence of TH fraction from Cabo Frio. The proportion of dead primary sporocysts
exposed to the different fractions was significantly higher than that of the secondary ones
(p<0.001), independently of the lineage (Taim / primary sporocyst – TH: 46.25%; H: 58%; S:
135
58%; Taim – secondary sporocys TH: 9%, H: 14.30%, S: 9%; Cabo Frio / secvondary sporocyst
– TH: 11.25%; H 3.2%; S: 2.9%. In the experiments using fluorescent markers, it was possible
to verify that primary sporocysts exposed to IDS – Taim showed labeling with the probe
Hoechst, Glycine max (soybean) lectin (SBA) and Alexa – Fluor 488, much more intense than
the exposed ones to IDS Cabo Frio. The secondary sporocysts exposed to IDS from Taim
and Cabo Frio snails presented a less marked labeling than the primary sporocysts, thus
allowing us to infer that S. mansoni at the secondary sporocyst stage is less damaged by IDS
components, even when IDS from a resistant lineage is considered. Nevertheless, labeling of
secondary sporocysts by three different probes was more intense when the parasites were
exposed to IDS from Taim lineage. Conclusions: IDS from B. tenagophila Taim is more active
against primary and secondary sporocysts than IDS from B. tenagophila Cabo Frio. Secondary
sporocysts are less susceptible to IDS from Biomphalaria than the primary ones, even when
related to a resistant strain, such as Taim lineage. This event may be due to the existence of
mimetism and/or molecular masking in the parasite at this evolutive stage. Financial support:
CAPES, CNPq, PRONEX - FAPEMIG
136
InvEStIgatIOn Of HYBRIdISM BEtWEEn BIOMPHAlARIA COuSInI and B. AMAZOnICA
tatiana Maria teodoro; Liana Konovaloff Jannotti-Passos; Omar dos Santos Carvalho; Roberta
Lima Caldeira
Centro de Pesquisas René Rachou/ Fiocruz, MG, Brasil
Introduction: Geographical distribution of schistosomiasis mansoni is directly associated
with the presence of susceptible snails of the genus Biomphalaria and the etiological
agent, Schistosoma mansoni. However, not all Biomphalaria species are susceptible to S.
mansoni. In Brazil, there are eleven species and one sub-species of molluscs of the genus
Biomphalaria, being three intermediate snail hosts of S. mansoni, B. glabrata, B. tenagophila
and B. straminea. The species B. peregrina, B. amazonica and B. cousini have been regarded
as potential hosts of the parasite. Although morphological studies of Biomphalaria species
identification have been widely carried out, differentiation between some species may be
difficult mainly due to phenotypic similarities, size of collected specimens and inadequate
fixation procedures. In these cases molecular techniques can help the classification. Previous
studies using phylogeny, morphological and molecular taxonomy, of Brazilian, Bolivian and
Colombian snails previously identified as B. amazonica, showed that some populations
studied were B. cousini instead B. amazonica. It was observed three different molecular
profiles which allowed to separate B. amazonica from B. cousini. The third profile showed the
association of the two before suggesting the possibility of hybrids between B. amazonica and
B. cousini. To study the presence of interspecific hybrids between these species, crosses were
done. We used the albinism factor (determined by a pair of recessive alleles), standardized
to Biomphalaria by Paraense (1955) as genetic marker. Besides, susceptibility studies were
done using the snails obtained from the crosses (hybrids). Methodology: To the crosses
experiments populations of B. cousini and B. amazonica kept in the René Rachou Research
Center and identified by PCR-RFLP were used. Considering the albinism as genetic marker,
four crosses were done using albino snails of B. amazonica and pigmented snails of B. cousini.
Each couple was kept in the same container for one week and after that they were separated.
Ten pigmented puppies obtained from each albino snail were individually identified by PCRRFLP to verify the profile of these descendents. To the susceptibility studies a total of 90
specimens of hybrids from Benjamin Constant, state of Amazonas, kept in the René Rachou
Research Center were exposed to 8 S. mansoni miracidia of the LE strain. As control exposed
to 8 miracidia 50 specimens of B. glabrata from Belo Horizonte were used. As control without
infection 10 specimens of B. glabrata and 10 specimens of hybrid from the same localizations
mentioned above were used. Susceptibility studies were done as described by Pellegrino and
Katz (1968) modified for Jannotti-Passos et al. (2008). If any specimen happened to die it
was examined by LS-PCR to detect the presence of S. mansoni. The specimens that survived
for 80 days after exposure without shedding cercariae were dissected and examined for
development stages of the schistosome. The specimens that shed cercariae were exposed
to light for 2 hours on alternative days until they died, and the cercariae were inoculated
in Swiss albino mice to verify the viability of these cercariae. This procedure was repeated
to SJ and AL S. mansoni strains. Results: The crosses between B. amazonica and B. cousini
showed that these species produce hybrids and that the molecular profile of these hybrids
was the same of the third profile obtained previously. Those results confirm the occurrence
137
of hybrids between B. cousini and B. amazonica. The hybrids proved to be susceptible to LE,
SJ and AL strains of S. mansoni with infection rate of 4.5%, 5.1% and 1.1% respectively. The
LS-PCR results are still being done. Conclusions: Hybrids can be considered a potential host of
the trematoda once they showed to be susceptible to all used strains of S. mansoni, shedding
viable cercariae. These results points to the risk of introduction of schistosomiasis mansoni
into new areas and more studies have to be done to better understand this hybrid.
138
COntROvERSIES REgaRdIng tHE SIgnIfICanCE Of tHE S0-CaLLEd “aPO” “aMEBOCYtE
PROdUCIng ORgan” In BIOMPHAlARIA GlABRATA
Samaly dos Santos Souza; zilton de araújo andrade
Centro de Pesquisas Gonçalo Moniz, Fiocruz, BA, Brasil
Introduction: Internal defense against microorganisms are performed in mollusks by a single
cell type: the hemocyte or amebocyte. These are phagocytic, mobile cells that can be found
wandering throughout the mollusk tissues, and circulating within the lymph. Their place
of origin in Biomphalaria glabrata has nowadays become a matter of controversy. Initially,
the hypothesis maintained by several Authors was that the amebocytes had a multicentric
origin, being formed from the cells present anywhere within the vasculo-connective tissue.
However, more recently it has been postulated that B. glabrata amebocytes are instead
formed within a central special organ, a correspondent of the vertebrate bone marrow, the
APO, “Amebocyte Producing Organ”, which is localized within the reno-pericardial area of
the mollusk. The initial main argument for the APO being considered as the locale of origin
for hemocyte production in B. glabrata was the finding of hyperplasia and mitoses in its
cells during the course of S. mansoni infection. On the other hand, other investigators have
described the so-called APO in Lymnaea truncatula as presenting a kidney-like structure,
typical of a filtration organ. Methodology: The present investigation was concerned with
a morphological analysis, with histological, immuno-histochemical, morphomentical, and
ultra-structural findings, from the so-called B. glabrata APO. In order to study its structure
and function in normal and S. mansoni-infected B. glabrata. Results: The “APO” was identified
as a collection of epithelial basophilic cells, disposed on one-cell-thick layer or in small
round collections, covering a small area of the pericardial surface in the reno-pericardial
region. Sometimes it vaguely resembled the epithelial component of the vertebrate juxtaglomerular apparatus, which reinforced the suggestion that such structure is related to
the kidney. During our studies mitoses were only occasionally found, either in normal and
infected mollusks. Also our quantitative studies failed to demonstrate the presence of APO
cellular hyperplasia, either in normal or schistosome-infected B. glabrata. Therefore, our
findings did not show evidences for the so-called APO to be considered as a central organ
for hemocyte production in B. glabrata. On the other hand, the multi-focal proliferation of
hemocytes was found in many other areas of the mollusk during S. mansoni-infection, as
initially suggested. Conclusions: On the other hand, several structural details from the “APO”
region in B. glabrata were found to be consistent with the suggestion that it is indeed a
filtration organ, more related to the kidney rather than to the bone marrow.
139
MaLaCOLOgICaL SURvEY Of BIOMPHAlARIA In tHE MUnICIPaLItIES Of “EStRada
REaL” In SOUtHWEStERn Of MInaS gERaIS StatE, BRazIL
Sandra Helena Cerrato tibiriçá; adalberto Mittherofhe; Milton f. Castro; adilson C. Lima;
Murilo gonçalves; Isabella O. Pinheiro; Corina C. freitas; Ricardo J.P.S guimarães; Omar
Santos Carvalho; Elaine Soares Coimbra
Universidade Federal de Juiz de Fora, MG - Brasil
Introduction: The increase in the practice of ecotourism and rural tourism in Minas Gerais
state, Brazil, involving primary contact with water coexists with the high and medium
prevalence of schistosomiasis present in various regions of the state. The State of Minas
aggregates most municipalities of the “Estrada Real” Project with great potential for
expanding tourism and rural ecotourism, because of the attraction represented by historicalcultural monuments, the diversity of topography, fauna and flora and bodies of water. Thus,
mapping the geographical areas vulnerable to transmission of schistosomiasis becomes a
task not only of health surveillance, but of encouraging productive sectors that drive the
economy in the state of Minas Gerais. This study aimed to identify and examine the genus
Biomphalaria molluks in the municipalities of the “Estrada Real” in the Southeast of Minas
Gerais State, Brazil Methodology: Mollusks of the Biomphalaria genus were collected from
August 2005 to May 2009, by technicians from GRS/JF. Catches were made in various parts
of the municipalities in order to cover the largest possible area with the aid of gloves,
tongs and dip nets. In all municipalities surveyed there are collections of water (lotic and
lentic), where there were characteristics would favor the occurrence of Biomphalaria genus
and the possibility of contact with human populations. At least, three water collections in
each municipality were surveyed. The points were georeferenced using a Global Position
System (GPS - Garmin Model II-12) and imported into a Geographic Information System
(GIS) to visualize the spatial distribution. The sites with the presence of Biomphalaria were
photographed with a digital camera. The captured snails were counted, transported in
plastic containers and kept in the laboratory until the time of identification. In general, 50%
of the specimens from each sample were separated for both morphological and molecular
study, and 50% examined for the presence of S. mansoni. The specific identification of the
mollusks was realized at the Parasitology Laboratory in the Federal University of Juiz de Fora
and the Entomology Laboratory of GRS/JF-SES/MG, according to the protocol established
by Paraense (1975, 1981). Some specimens were sent to the Laboratory of Helminthology
and Malacology at the René Rachou Medical Research Center/Fiocruz for confirmation of
species by the technique of Polymerase Chain Reaction-PCR (Vidigal et al., 2000). Results:
The research for Biomphalaria was realized in 36 municipalities, of which 19 are part of the
“Estrada Real” Project and the others are within the coverage area . 30 municipalities were
positive for snails of the genus Biomphalaria, and in only 6 the presence of mollusks not
observed. Six Biomphalaria species were found: B. glabrata, B. tenagophila, B. straminea
B. peregrina, B. occidentalis and B. schrammi. The B. tenagophila species was found in
16 municipalities; followed by B. peregrina, in 15 municipalities and by B. glabrata, in 13.
A total of 3,622 specimens of mollusks were collected, all negatives for cercariae and/or
sporocysts of the S. mansoni Conclusions: Considering the occurrence of schistosomiasis in
the State of Minas Gerais and the social-economics repercussions that involve the “Estrada
140
Real” Project, this work focuses on the subject of the study in vulnerable water collections
of the presence of Biomphalaria mollusks and the need of sanitary and education measures
integrated with participating communities, epidemiological surveillance and tourism
sectors. This is a pioneering work in order to search for Biomphalaria in municipalities of the
“Estrada Real” using the location of specimens by geographic coordinates/GPS. The presence
of Biomphalaria snails, intermediate hosts of S. mansoni, in association with practices of
ecotourism, rural tourism and water sports can support the expansion of the transmission
area of schistosomiasis in Minas Gerais.
141
RISK aREaS Of SCHIStOSOMIaSIS nEaR tHE BRazILIan BORdER IntO tHE URUgUaY
RIvERS BaSInS In tHE PROvInCE Of CORRIEntES, aRgEntIna
Maria Jf Rea; C. Edgardo Borda; Osvaldo d. Benitez; Luis a. Mosqueda
Centro Nacional de Parasitología y Enfermedades Tropicales, Facultad de Medicina, Unne,
Corrientes, Argentina
Introduction: The southern limits of the geographical distribution of schistosomiasis in
Brazil are the states of Paraná, Santa Catarina, and Rio Grande do Sul border the Argentina.
Among the Biomphalaria species that occur in Brazil, three are regarded as intermediate
hosts of Schistosoma mansoni: Biomphalaria glabrata, B. tenagophila and B. straminea.
In our previously published reports we emphasized the possibility for the introduction
of schistosomiasis in non-endemic ar¬eas in Argentina into the basins of the Paraná and
Uruguay Rivers. It was demonstrated that life cycle S. mansoni is closed with planorbid (B.
tenagophila) and rodents (Holochilus braziliensis) that live in the same ecological niche.
Determining the distribution of susceptible and compatible Biomphalaria species provides
information of possible areas where sporadic outbreaks of schistosomiasis could appear.
The aim of the present study was to identify the Biomphalaria species of the province
of Corrientes and assess their susceptibility and compatibility to S. mansoni infection, as
potential intermediate hosts. Methodology: This province is located in the humid subtropical
region of Argentina northeast in the border with the South of Brazil. Uruguay River is the
natural frontier between Argentina and Brazil. Survey has been conducted over four years
(2006-2009) in six Departments of the eastern region in the Uruguay River basins, where
close and frequent human contact with the water was observed. The collection areas are
included between the 28-30° parallels and the 56-59° meridians. A total number of 728
specimens were collected in 21 freshwater locations. Subsequent examination using the
light exposure test in the laboratory determined whether the specimens were positive for
S. mansoni. The molluscs collected were sent to our laboratory to obtain their F1 progeny.
Studies of shell morphology and the internal anatomy of planorbids were conducted to
identify species. Results: The following species were found and identified: Biomphalaria
tenagophila, B. straminea and B. orbigny. The B. tenagophila populations were obtained from
16 water bodies of five Departments. They were collected from seven locations in Mercedes,
four in Paso de los Libres, two in Curuzú Cuatiá and Alvear and only one in San Martín. The
other species were identified in only one place, B. straminea in Santo Tomé and B. orbigny
in Mercedes. All field-collected molluscs were determined to be S. mansoni-negative.
Groups of 100 F1- snails (4-8 mm) of B. tenagophila collected in 13 locations were exposed
with 10 miracidia/mollusc of the SJ2 strain from São José dos Campos (state of São Paulo),
Brazil, more adapted to B. tenagophila. This strain is routinely maintained at the Cenpetrop
laboratories by passages through sympatric B. tenagophila and hamster or Swiss albino
mice. As an infection control, we included 100 B. tenagophila specimens from São José dos
Campos, infected with 10 miracidia /mollusc of the same strain, which had been kept in the
Cenpetrop laboratories. The populations of B. tenagophila used for the susceptibility tests
from eigth collection areas, showed no evidence of either S. mansoni cercariae or sporocysts.
However, the snails from five other localities were susceptible. Specifically, they were
collected in Mercedes, from two streams (Sarandi, Arazá) and one dam (Curupicay) in Paso
142
de los Libres from one dam (Mirungá) and in Curuzú Cuatiá from one stream (Ibabiyú). The
infection index varied between 2- 36% and the precercarial period varied from 31-62 days.
The mortality index in the precercarial period was 18%. We developed a quantitative method
TCP/100 index (degree of compatibility between the Schistosoma and its intermediary host)
proposed by Frandsen. Three populations from Mirungá, Arazá and Curupicay were not very
compatible to the SJ2 strain (Class I) with an index less than 10,000 cercariae. The snails
from Sarandí stream showed an index of 13,572 (Class II, poorly compat¬ible). Meanwhile,
those snails from Ibabiyú were compatible with a higher index of 50,602 (Class III). In the B.
tenagophila controls, 42% of the exposed snails were infected (index of 6,112, Class I, not
very compatible). The prepatent period average was 44 days. Conclusions: Investigations on
experimental infection using B. tenagophila have shown that they are potential hosts for the
trematode in these regions. Workers arrive to these places and they settle down precariously
in the periphery of urban centres in proximities of superficial waters. With these results, in
addition to environmental and social changes that took place in the Uruguay River basins
(the increasing numbers of dams, irrigation channels and the scrub vegetation along the
shoreline) during the last two decades the probability of schistosomiasis introduction in this
region exists.
143
PaRaSItIC CaStRatIOn In BIOMPHAlARIA GlABRATA dURIng tHE PRE-PatEnt and
PatEnt InfECtIOn WItH SCHISTOSOMA MAnSOnI
Marta Julia faro dos Santos Costa; Mariana Perazzini; Lygia dos Reis Corrêa; Clélia Christina
Mello Silva; arnaldo Maldonado Junior
Escola Nacional de Saúde Pública, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, RJ - Brasil
Introduction: Schistosomiasis is one of the most widespread parasitic diseases in tropical
countries according to the WHO, seventy-four countries are considered endemic. In Brazil
B. glabrata is intermediate host species of schistosomiasis more susceptible, presents a
wide geographic distribution, being responsible for many outbreaks active this trematode.
It is widely accepted that trematode parasites adversely affect the fecunditity of their
intermediate molluscan host. In B. glabrata snail, the inhibition of egg-laying has been shown
to begin at the 4th week after infection by Shistosoma mansoni and castration concides with
the onset of cercarial appearance from week 6 post-posture. Complete suppression of egglaying is referred to as castration and host castration is of phenomenon evidente in many
parasitic relationships.The process has been related to parasite induced gonad damage
and involve alteration in the functioning of host accessory glands or endocrine systems. In
the literature there is a lack of information on the behavior of the species of Biomphalaria
and their reproductive aspects in infection, since the majority of studies used molluscs not
infected. The objective of this work was to observe the alterations reproductive (fecundity
and fertility) of Biomphalaria glabrata infected with Schistosoma mansoni in periods pre and
patent of infection. Methodology: The molluscs used in this work are from Ressaca, Minas
Gerais and were infected with Schistosoma mansoni (BH strain). It was employed 60 molluscs
after reaching sexual maturity of approximately 8-10 mm in diameter. Thirty specimens were
individually exposed to 5 miracidia of Schistosoma mansoni and 30 were used for the control.
The parameters analyzed were the fecundity and fertility. The molluscs were exposed to
light from the 30o day to verify the release of cercariae and the positives molluscs were
maintained in aquaria separately. Weekly, the survival rate and the variation of growth
were measured through the development of the shell. These observations were carried
out until sixty and two days from the exhibition. The results of the fecundity and fertility
determinations were evaluated by the ANOVA, with the Tukey-Kramer test for comparation
of the means (p<0.05). Results: The population of infected B. glabrata showed 50% positive
and 100% survival. Significant differences were observed among the analyzed only in the
pre-patent in the snails exposed to. We observed an average growth rate, which ranged from
10.6 and 17.0mm for the control and 10.6 and 17.7 mm for the exposed. The positive snails
were measured from day 35o ranged from 15.0 and 18.0mm. In the pre-patent period it
was found that the number of egg masses/snail showed a significant difference in the group
exposed to infection during periods of 7-14 days (q = 5.3 and p <0.05) and 7-28 (q = 5.3 and
p <0.05) days. Regarding the number of eggs/snail and eggs/egg mass was no significant
difference of these two parameters simultaneously in periods 7-14 (q = 7.1 and p <0.01) and
7-21 (q = 8.3 p <0.001). As for the patent period was not significant difference between the
reproductive parameters over the period. The positive snails that shed cercariae, oviposition
ceased in the early periods of elimination of the larvae (35-50 days). In the period 55-62
days of infection, the snails held positive attitudes with production of 1.5 eggs per clutch,
144
but not hatched snails. Fertility number of hatched snails was proportional to the number
of eggs per clutch over the two study periods in both the control group in terms of the
above. In the pre-patent period was observed significant difference in the intervals 7-14 (q
= 6.8 and p <0.01), 10-17 (6.7ep q=0.01) and 14-31 (q = 6.9 and p<0.01). Conclusions: The
exposed snails grew more than the control group and the positive grew faster than the two
groups. In this study, we found that snails exposed and control standards were coming for
reproductive activity in the pre-patent and patent infection. On the other hand, different
behaviour was observed for the molluscs positive. After the cercariae began to emerge at 35
days, fecundity and fertility almost completely stoped. The reduction in fecundity associated
with the end of larval development of S. mansoni coincides with the depletion of energy
storage, as well as glycogen storage in the gland digestive. It was observed an interruption
of the activity of molluscs positive ovipositor, which may be associated with biochemical
alterations (castration indirect) and with possible injuries ovotestis (castration direct).
145
MaIntEnanCE Of HUMan and WILd CYCLE Of SCHISTOSOMA MAnSOnI In LaBORatORY
Marjane Soares ferreira; andiara garcez de Souza Silva; Selma Patrícia diniz Cantanhede;
Luciana Patrícia Lima alves; Hallyne davinck Mesquita Moreira; nêuton Silva-Souza; Juliana
araripe gomes da Silva
UFPA, MA - Brasil
Introduction: The trematode Schistosoma mansoni, the causative agent of schistosomiasis in
Brazil, presents a complex life cycle, which alternates phases in vertebrates hosts and phases
in invertebrate host, molluscs from the gender Biomphalaria (Gastropoda: Planorbidae). In
peripheral neighborhoods of São Luís - MA these snails can be found in large quantities, many
of them capable of transmitting the disease. These snails, as intermediate hosts, also inhabit
the fields in the Baixada Maranhense, coexisting with the wild rodent of semi-aquatic habit
Holochilus brasiliensis, another definitive host, other than man, and well adapted to the
cycle of S. mansoni, acting as a source of disease dissemination in the region. Methodology:
In the laboratory were kept the two cycles of Schistosoma mansoni: the human, using as
an experimental model the mice Mus musculus and the wild cycle, with the use of rodents
Holochilus brasiliensis, from the city of São Bento, Maranhão State. For the human cycle
were collected 50 snails Biomphalaria glabrata in peripheral neighborhoods of São Luís MA, referred to the Human Parasitology Laboratory (HPL), State University of Maranhão. This
year lots were collected in Bragança - PA, for genetic analysis. During five weeks, the snails
were examined individually as to positivity by eliminating cercariae. Negative snails were
used for mass infection with miracidia obtained from eggs of S. mansoni, found in human
feces positive for the parasite. These samples were obtained from a Public Health Laboratory
of São Luís - MA, or feces of infected mice in previous cycles. Depending on the amount of
negative snails, each week were put lots 5-10 snails to infection. From the 25th day after
exposure to miracidia, new positive tests were performed, in order to obtain cercariae
infection of mice. We used 6-10 Mus musculus, with 30 days after birth, from the vivarium
UEMA. Subsequently, the mice were infected percutaneously, partly submerging them for
1 hour in a solution of 200 ml of distilled water containing cercariae previously counted.
From the 30th day after infection, the stool tests were initiated in mice by weekly analysis by
Kato-Katz, always made in triplicate. Finally, the total number of positive rodents was kept a
group of rodents for up to three successive cycles of creation, and extracted the other adult
worms, which are used for different works in the laboratory. To mount the wild cycle we used
the same procedures of the human, with the difference of not using human feces and work
with wild rodents. For this cycle, Biomphalaria glabrata snails were collected in São Bento,
Maranhão State. Males and females of Holochilus brasiliensis were also captured in the same
location for subsequent reproduction in the laboratory. Was made so, test positive of rodents
for schistosome infection and other intestinal parasites by the Kato-Katz. Rodents positive for
S. mansoni were maintained in laboratory for feces collect in order to infect the snails and
eventually proceed with the infection of H. brasiliensis until then negative. The collection
of adult worms followed the same protocol established for the human cycle. Results: From
2006 to 2010 were carried out successive cycles, both human as wild Schistosoma mansoni
successfully, and the monitoring and observation occurred in the laboratory. During this
period there were, per year, two human and two wild cycles complete. Compared to the
146
first cycles performed in 2006, on the 50 snails collected for the assembly of the first human
cycle, we obtained a percentage of only 20%-30% of infected snails in the three batches of
10 (each batch a week) and some succumbed to the infection. However, infection of mice
was 100%. For a first wild cycle obtained a higher percentage of 60% to 70% for the snails,
with also 100% of infection for wild rodents. The year 2009 was distinguished by having the
highest rates of infection, up to 80% in snails to both cycles, continuing with the percentage
of 100% of infection both for mice as for wild rodents. Conclusions: From 2006 until this year
have been successfully kept successive cycles of S. mansoni in the laboratory, both human
cycles, the wild. This ability to maintain cycles under conditions somewhat different nature is
facilitated by considerable adaptation and resistance of experimental models Biomphalaria
glabrata and Mus musculus to the laboratory conditions.
147
EvaLUatIOn Of tHE EffECt Of tHE aSSOCIatIOn IMMUnIzatIOn-CHEMOtHERaPY
On SCHISTOSOMA MAnSOnI InfECtIOn.
Ludmila zanandreis de Mendonça; Patricia Martins Parreiras; tatiane teixeira de Melo; neusa
araújo Pereira; Juliano Michel araújo; flávia fernanda Búbula Couto; ana Carolina alves de
Mattos; Paulo Marcos zech Coelho; Cristina toscano fonseca
Centro de Pesquisas René Rachou/ Fiocruz, MG - Brasil
Introduction: Schistosoma is a parasite which causes chronical infection in more than 200 million
people of developing countries. Schistosomiasis is the most important human helminthic
infection in terms of morbidity and mortality. Treatment of infected individuals with effective
and safe drugs as Oxaminiquine and Praziquantel led to morbidity control, however although
mass treatment with drugs decreases the prevalence and the infection intensity, the large
extension of endemic areas and constant reinfections together with poor sanitary conditions
makes chemotherapy an ineffective control strategy. For these reasons it is clear that there
is a need for new alternative estrategics fo controlling schistosomiasis. Currently, several
researchers believe that the combination of immunization and treatment could be effective to
control the disease, since treatment effectiveness is associated with host immunity. Association
could reduce drug dose (decreasing the side and toxic effects); increase the therapeutics index;
reduce the treatment cost and could be effective on resistant strains. In studies conducted by
our research group, the schistosomulum tegument (Smteg) was able to induce partial protection
in the murine model. The tegument is critically involved in host-parasite interactions. Regarding
immune evasion, several mechanisms including antigenic mimicry, proteolytic degradation
of “attacking” host proteins, rigid biophysical membrane properties and a rapid tegumental
membrane turnover takes place in the tegument, suggesting its importance to parasite survival.
In this work we evaluated whether the association immunization-chemotherapy increases
Praziquantel (PZQ) efficacy against the parasite. Methodology: C57BL/6 mice were vaccinated
with three doses (200µL/mice) of PBS + CFA in the 1st dose and IFA in subsequent doses or
Smteg + CFA/IFA (25µL/mice) with a 15-day-interval among the doses. Thirty days after the third
dose, mice were infected with 50 S. mansoni cercariae, LE strain. Forty-five days after infection
mice were treated with 200mg/kg PZQ, and 20 days after treatment the animals were perfused
in order to assess the protection level. Three days before perfusion, a stool examination was
carried out using two methodologies: HPJ and Eclosion of Miracidia. The liver and intestine
were collected after perfusion, weighed and digested with 10% KOH, and the number of eggs
present in these tissues was determined under optical microscopy. During the experiment, the
blood of these animals was collected in order to evaluate the production of specific antibodies
by ELISA. Results: No significant difference in the number of adult worms was observed between
immunized mice (Smteg) and immunized/treated mice (Smteg + 200mg/kg PZQ) although
in not immunized and treated mice a significant reduction on worm burden was observed,
when compared to infected and non-treated mice, suggesting that the antibodies induced by
Smteg immunization are blocking the drug target in adult worm. Conclusions: Our preliminary
results indicate that the association of immunization with schistosomulum tegument (Smteg)
and Praziquantel treatment do not increase drug efficiency, may even inhibit its action.
Other “in vivo” and “in vitro” studies are being performed in order to clarify these results.
148
EvaLUatIOn Of tHE taQMan REaL tIME PCR aS an aLtERnatIvE fOR dEtECtIOn
Of SCHISTOSOMA MAnSOnI EggS In fECES aftER tHE COnCEntRatIOn HELMIntEx
MEtHOd
Candida fagundes teixeira; aline Cardoso Caseca volotão; Carlos graeff teixeira
PUCRS, RJ - Brasil
Introduction: Classical parasitological methods are not sensitive enough to detect most of
the infected individuals in low intensity transmission areas like in Esteio, Rio Grande do Sul,
the southernmost Brazilian State. Helmintex is A novel diagnostic method for isolation of
Schistosoma mansoni eggs in feces. It’s a concentration method with sequences of spontaneous
sedimentation, sieving and a final step where eggs are isolated through interaction with
paramagnetic beads, and the sediments retained at the wall is collected and examined under a
microscope The aim of the present study was to evaluate TaqMan Real Time PCR as an alternative
for microscopic analysis as the detection procedure for Helmintex. Methodology: Primers and
probe targeting the cytocromo-C oxidase subunit 1 gene were designed for amplification DNA de
Schistosoma mansoni. Four procedures for DNA extraction were evaluated. S. mansoni eggs were
collected from fecal sediments processed by Helmintex and suspensions in distilled water were
prepared containing 1, 10, 20, 40 and 80 eggs. As specificity control, a fecal sediment was also
prepared from heavily parasitized samples with Ascaris lumbricoides, Taenia spp. and addition
of Fasciola hepatica eggs collected from adult worms. S. mansoni DNA was extracted with Fast
DNA Kit (MP Biomedicals) from 10 male worms and also from 1000 eggs, for determination of
a standard curve for estimatives of DNA concentration. The standard curve was obtained after
estimation of DNA through readings of serial dilutions at base 10 (from 100 ng to 10 fg) with
Nanodrop Spectophotometer ND 100. Results: The standard curve for DNA concentration from
10 worms had a detection limit at 100 fg and Ct = 35,5 ± 1,31 (average ± standard deviation). At
the ideal conditions DNA was amplified from samples containing up to 10 ng and Ct = 20,94 ±
0,58. Reprodutibility was better with the latter than with the former preparation. The standard
curve obtained from the sample with 1000 eggs, detection limit was 10 pg and Ct = 34,2
± 1,24. Amplification was detected up to 10 ng and Ct = 23,7 ±0,46. DNA was amplified from
all the spiked egg suspensions, including the samples with only 1 egg. Illustra Tissue and Cell
GenomicPrep Mini Spin Kit (AMERSHAN) presented with the best extraction performance, with
DNA detected in all samples, Ct = 29,42 for 1 egg and Ct = 22,82 for 80 eggs. QIAmp DNA Stool
Handbook (QIAGEN), also detected DNA in all samples, Ct = 34,55 for 1 egg and Ct = 25,06 for 80
eggs. Adapted protocol as described for Pitcher (1989) e Silva (1999) amplified from 1, 10, 20, 40
and 80 eggs, with respective values of Cts: 41,07, 34,08, 32,87 e 80 35,94. Fast DNA Kit showed
amplification from 10 (33,99), 40 (33,68) e 80 (35) eggs. DNA was not amplified with control
samples containing DNA from A. lumbricoides, Taenia spp. e F. hepatica. Conclusions: Real Time
PCR showed a satisfactory performance to amplify DNA from samples with very small numbers
of eggs. Specific amplification appears to occur when sequences of cox 1 gene are used for S.
mansoni DNA detection, as proposed by Hove (2008). These preliminary results show a potential
role for Real Time PCR for detection of eggs in feces after the concentration Helmintex method,
what may further increase its sensitivity detection limit estimated as 1.3 eggs /gram of feces. This
improvement will add support for control measures of schistosomiasis in low intensity areas.
149
MOLECULaR anaLYSIS Of tHE OvIPOSItORY BEHavIOR Of MURInE SCHIStOSOMIaSIS
Eduardo finger; thaissa Melo galante Coimbra; Érika Lopes fernandes; débora Cristina
Rosseto garotti
Santa Casa de São Paulo, SP - Brasil
Introduction: The gold standard for the diagnosis of schistosomiasis is the direct
demonstration of parasite eggs in stool by the Kato-Katz technique (KK) however, this
technique’s high sensitivity threshold demands a minimum of 24 eggs/gram of feces for a
diagnosis, something that makes it unsuitable for situations of low oviposition such as low
endemicity areas or recent infection. In these situations, more sensitive methods like the
polymerase chain reaction technique (PCR) are warranted. Due to the singular difficulties of
purifying amplifiable DNA from feces, PCR has never become a mainstream technique for the
diagnosis of schistosomiasis and therefore, little is known about how it compares to KK in
detecting oviposition along the chronology of Schistosoma infection. This work aims to study
oviposition in experimental murine schistosomiasis through the PCR and KK techniques, to
compare their efficacy detecting the start of oviposition after infection and its cessation after
treatment. Methodology: Feces from mice infected with S. mansoni were collected weekly
beginning on the 2nd week-post-infection (wpi). On 7th wpi, mice that tested positive for
schistosomiasis through PCR were divided in 2 groups, one was treated with oxamniquine
and the other was left to follow the natural course of the infection. Feces continued to be
collected until all treated mice tested negative for the presence of Schistosome genetic
material in stool as detected by PCR. Subsequent to PCR testing, all stool samples underwent
examination through KK and the results were aligned along PCR results for comparison.
Results: Current partial results indicate that schistosome genetic material can be detected on
stool after 4 weeks of infection, the same time frame currently attributed to KK. Conclusions:
At least when tested on a weekly basis, PCR and KK were equally effective at determining the
start of oviposition. In order to better verify the effectiveness of each method, samples must
be collected and tested more frequently around the time of the start of oviposition between
the 3rd and 4th wpi.
150
IdEntIfICatIOn Of SCHISTOSOMA MAnSOnI antIgEnS CandIdatES tO BE USEd In
tHE dIagnOSIS Of SCHIStOSOMIaSIS.
gardênia Braz figueiredo de Carvalho; Cintia Maria gonçalves da Silva; Lucila gonçalves
grossi Pacífico; Cristina toscano fonseca
Centro de Pesquisas René Rachou/Fiocruz, MG - Brasil
Introduction: Schistosomiasis is the most important human helminthic infection in terms
of public health and affects more than 200 million people in 74 countries worldwide. The
diagnosis of the disease is a key for the disease control. Currently the diagnosis is performed
by the Kato Katz method that has limited sensibility in individuals with low parasitic burden.
A diagnosis assay able to identify infected individuals with low parasitic burden would
provide the necessary data to design effective control strategies and to determine the
efficacy of control interventions. In this work we have performed a bioinformatics analysis of
Schistosoma mansoni genome in order to identify promising antigens to be used as antigens
in a serological diagnostic assay for schistosomiasis. Methodology: The antigens were
selected using schistodb database (www.schistodb.net) based on the gene expression in
the different stages of the parasite life cycle (schistosomulum, lung schistossomulum, adult
worm and egg,) in the definitive host. The ESTs sequences corresponding to antigens was
find in pubmed. Numerous genes were identified, and in silico analysis were performed to
select candidates (signal peptide, transmembrane domain, predicted cellular localization and
percentage of homology to human protein). After that, the sequences found were submitted
to B cell epitope prediction using software “bcpred” (ailab.cs.iastate.edu/bcpreds) and
protein were also submitted to 3D computational modeling. We performed the alignment
of the selected genes with the human homology sequences and compared the location of
the B cell epitopes predicted between Schistosoma and human proteins. Besides, genomic
DNA was extracted from S. mansoni adult worms using wizard DNA purification systems
(promega) for gene amplification by PCR to be cloning in the plasmid (pET21a) that will be
used for gene expression. In parallel we infected 130 swiss mice with 25 cercariae (LE strain)
and blood sample was collected to be used in the serological tests. Mice were divided into
two groups: group 1 (50 mice) and group 2 (80 mice) for oxaminiquine treatment. At the
end of the protocol all mice will be perfused for worm burden determination. Results: At the
first analysis in Schisto db database 72 sequences were found based in gene expression in
the different stages of the parasite life cycle (schistosomulum, lung schistossomulum, adult
worm and egg) and the presence of a signal peptide. Following the analysis of the predict
cellular localization and the percentage of homology to human proteins only 10 sequences
were selected based on its localization on the membrane or its prediction to be secreted and
its low homology to human protein. These 10 sequences were submitted to B cell epitope
prediction and based on the number of B cell epitopes and its presence only on S. mansoni
proteins, 6 candidates to be tested for the serological diagnosis of the disease were selected.
These proteins will be produced as recombinant antigens and evaluated regarding their ability
to be recognized by sera for infected mice. Conclusions: Screening candidates to be tested for
diagnostic of schistosomiasis using in silico tools led to identification of promising antigens
that can be now produced as recombinant protein and evaluated by means of serological tests.
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EvaLUatIOn Of HEPatIC fIBROSIS In SCHIStOSOMIaSIS BY BIOLOgICaL SERUM
MaRKERS
tibério Batista de Medeiros; ana Lúcia Coutinho domingues; Edmundo Pessoa de almeida
Lopes; ana virgínia Matos Sá Barreto; Clarice neuenschwander Lins de Morais; Silvia Maria
Lucena Montenegro; José Roberto Maciel Martins
Departamento de Medicina Clínica da UFPE, PE - Brasil
Introduction: Introduction: Schistosomiasis is an infectious disease with more than 200
million people affected worldwide. In Brazil, between 6 and 7 million are infected by
the Schistossoma mansoni. Many organs and tissues are affected by the disease, but the
most characteristic damage is the fibrosis that occurs around portal hepatic branches,
known as periportal fibrosis. The understanding and following up of this fibrosis are very
important for a better approach to these patients. Actually, upper abdominal ultrasound
is the most used diagnostic tool to access the periportal fibrosis, but is not available in all
endemic areas and is subject to inter-and intraobserver variation that is why new methods
are under development to evaluate the hepatic fibrosis in schistosomiasis. To avoid the
mentioned limitations of ultrasound, serum markers of hepatic fibrosis have been evaluated
in different hepatic diseases with good results so far, but in schistosomiasis, there are not
many studies available yet. This study aims to correlate serum markers such as aspartatoaminotransferase/alanino-aminotransferase (AST/ALT) ratio, gama-glutamiltranspeptidase
(GGT), platelets, immunoglobulin G (IgG), hialuronic acid (HA) and APRI (ratio AST/platelet)
índex with different patterns of periportal fibrosis established by ultrasound. Methodology:
Methodology: Between march and December of 2009, 122 patients were evaluated
consecutively in schistosomiasis ambulatory of the Hospital das Clínicas of the Federal
University of Pernambuco. There were 12 patients controls from endemic areas and no
fibrosis and 110 with any degree of periportal fibrosis by ultrasound. All patients had blood
sample drawn for measurement of AST, ALT GGT, blood cell count with platelets, HA, IgG and
in the same day abdominal ultrasound was performed always by the same physician. Hepatic
schistosomiasis was accessed accordingly to Niamey classification: A- no fibrosis, B- doubtful
fibrosis, C – peripheral fibrosis, D - central fibrosis E-advanced fibrosis and F- very advanced
fibrosis. Patients were classified into 3 different groups: control, patterns A an B (group 1),
mild periportal fibrosis, patterns C and D (group 2) and advanced fibrosis, patterns E and F
(group 3). To identify the best cutoff of GGT, IgG, platelets, AST/ALT ratio and APRI index, to
diagnose fibrosis and after to differentiate mild to advanced forms, ROC (receiver operating
characteristic) plot was built. Results: Results: Platelet count had significant negative
correlation to the different periportal fibrosis patterns which means that the more advanced
stages of fibrosis had lower platelets count as seen in group 1 (257.833 ± 74.215) mm3 group
2 (158.355 ± 79.729) mm3, and group 3 (96.430 ± 45.355) mm3. The platelet count accuracy,
represented by area under curve “ROC”, to identify patients with periportal fibrosis was
0,921, being the value of 171.000 mm3 the best cutoff with 80% of sensibility and specificity
of 91.7%. Serum levels of GGT, expressed in upper normality limit were in group 1 (0,61
± 0,29), group 2 (1,29± 1,06), and group 3 (2,54± 1,81). The accuracy was 0,85, being the
value of 0,84 of the upper normality limit the best cutoff to differentiate patients with or
without fibrosis with 74,6% and 83,3% sensibility and specificity, respectively. The medium
152
serum levels of IgG were 1236 mg/dl (± 368) in group 1, 1338 mg/dl (± 379) in group 2 and
1647mg/dl (± 242) in group 3. The value of 1542mg/dl was the best cutoff to identify those
with advanced fibrosis with sensibility of 57,3%, specificity 91,7% and accuracy of 0,736.
Serum levels of HA in the no fibrosis group were 23,9 mcg/l, mild fibrosis was 51,9mcg/l, e
64,3mcg/l those with advanced fibrosis. To identify the best cutoff for patients with fibrosis
the ROC plot showed a serum value of 27,8 mcg/l with sensibility of 78,2% and specificity of
83,3,7%. APRI index showed progressive values according to the groups: 1 (0,26 ± 0,11), 2
(0,92 ± 0,98) and 3 (1,58 ± 1,40) and accuracy to identify fibrosis was 0,930 , and the value of
0,349 as the best cutoff with sensibility of 90% e specificity of 83,3%. The evaluation of AST/
ALT ratio showed no correlation with the groups studied. Platelets and APRI index were the
best predictors of fibrosis when used singly. Conclusions: Conclusion: This study observed
that platelets, IgG, GGT, HA and APRI índex had correlation with periportal fibrosis diagnosed
by ultrasound and are promising serum markers of fibrosis in mansonic schistosomiasis.
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an OUtBREaK Of aCUtE SCHIStOSOMIaSIS In tHE SOUtHEaSt Of BRazIL: II. CLInICaL
aSPECtS
José Roberto Lambertucci; Sandra Costa drummond; Izabela voieta; alba Otoni; Bruna assis
Chaves; Pedro Henrique Prata; Pedro Paulo nunes Pereira; thiago Cardoso vale; Leonardo
Campos de Queiroz; Paloma fonseca; Carlos Maurício de figueiredo antunes
Faculdade de Medicina da UFMG, MG - Brasil
Introduction: Acute schistosomiasis has been reported in Brazil since the description of the
disease by Pirajá da Silva in 1908. The morbidity of acute schistosomiasis, however, has been
largely underestimated and in most reports it has been described as a self limited disease
that should spontaneously evolve into the chronic stages without serious consequences.
Here we outline an outbreak of acute schistosomiasis mansoni in a low transmission area
of Minas Gerais state in Brazil in which five patients were admitted to hospital with severe
complications of the disease. Methodology: Forty one individuals who bathed in a swimming
pool filled in with untreated natural water in an country estate in the outskirts of São João
Del Rei, from December 2009 to February 2010, were submitted to clinical, laboratory and
ultrasound examinations. A standard protocol was designed specifically for this outbreak
investigation containing socio-demographic, clinical, laboratory and ultrasound data.
Two stool examinations by the Kato-Katz method and by a sedimentation technique were
performed in all patients. Clinical examination was done by one of us (JRL) with particular
attention to abdominal palpation. The right hepatic lobe was examined on the anterior
axillary line and the left lobe on a line passing through the xiphoid appendix. The spleen
was palpated and measured under the costal margin with the patient in dorsal decubitus
during deep inspiration. For ultrasound, a Medison Sonoace 1500 3.5 MHz apparatus was
used. A 10 ml of venous blood was collected and stored in a freezer at -20ºC for serological
examination. The medical files of five patients admitted to hospital were reviewed and
clinical details of the cases during hospitalization were obtained. Biomphalaria glabrata
snails were identified in the swimming pool and in the neighboring stream waters. Results:
Patients age ranged from 2 to 82 years (mean: 33.5±19.8; median: 39) and 21 were male
(51%). Fifteen patients (36.6%) passed S. mansoni eggs in the stools. Sixteen patients (39%)
presented symptoms: cercarial dermatitis in 13 (31.7%), cough in 10 (24.4%), diarrhea in
9 (22.0%), fever in 7 (17.0%), bloody stools in 7 (17.0%), lower limb weakness in 1 (2.4%).
Physical examination revealed hepatomegaly in 16 (39%) and a palpable spleen in 6 (14.6%).
Ultrasound showed the following alterations: hepatomegaly in 13 (31.7%) - 6 had both right
and left liver lobe enlargement -, splenomegaly in one (2.4%) and periportal lymph nodes
in 10 (24.4%). Five patients were admitted to hospital with severe manifestations of the
disease: three with pulmonary involvement, one with severe gastrointestinal symptoms and
one with myeloradiculopathy. In two patients the disease evolved with fever for more than
30 days fulfilling the criteria for the diagnosis of fever of undetermined origin. One out of
the 3 with pulmonary involvement had micronodules in both lung fields confirmed by a CT
scan of the chest. The other three had no x-ray alterations. All patients admitted to hospital
had eosinophilia. In a 27-year-old female the disease evolved with lower limb pain and
paresthesia followed by urinary and fecal retention. Cerebrospinal fluid revealed pleocytosis
with an increased number of mononuclear cells (70 cells/mm3) and proteins (1,440 mg/dl).
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Magnetic resonance imaging showed a lesion at the distal portion of the conus medullaris,
hyperintense in T2-weighted sequence with heterogeneous contrast enhancement.
Her husband and daughter also presented fever, vomiting and myalgia, and positive
parasitological stool examination for S. mansoni ova, but with no neurological involvement.
She was treated with steroids and praziquantel and presented complete recovery 4 months
after starting treatment. Except for one patient who refused treatment, the others received
praziquantel (50 mg/kg/body weight, single dose). Conclusions: Physicians should be aware
of acute schistosomiasis occurring in low transmission areas. Severe manifestations of the
disease described in this outbreak may be explained by the duration of exposure and/or
intensity of the infection; underestimation of morbidity in previous reports is also likely.
155
PROLOngEd COURSE Of SCHIStOSOMaL PULMOnaRY HYPERtEnSIOn
Rita de Cassia dos Santos ferreira; ana Lúcia Coutinho domingues; angela Pontes Bandeira;
amanda Medeiros gomes da Silva
Introduction: The aim of the present study was to report the evolution of nine patients
undergoing treatment for schistosomal liver fibrosis, at Hospital das Clínicas – Universidade
Federal de Pernambuco that presented with pulmonary hypertension by transthoracic
Doppler echocardiography (pulmonary arterial systolic pressure - PASP > 35mmHg) during
a previous prevalence study carried out three years before. Methodology: All nine patients
or their family were called by phone and after informed and written consent they were
submitted to medical histories, physical examination and functional classification as I, II, III
and IV. Abdominal ultrasonography, transthoracic echocardiography to estimation of PASP
and spirometry were performed. Data of their cardiac catheterization and six minute walk
test when performed were compilled. Results: Two of the nine patients enrolled were dead
in 2009 year: two females, one 31 years old (PSAP 79mmHg, functional class II) and the
other 61 years old (PSAP 126mmHg, functional class IV). The fisrt one was treated with
calcium canal blocker and the other one with the phosphodiesterase inhibitor and bosentan.
However, these two patients had early diagnosis of pulmonary hypertension 5 and 8 years
ago. One patient was not found and another couldn’t be present at that clinic, but was alive.
Three patients were not submitted to right heart catheterization because their disease were
mild (functional class I) and their treatment was not indicated at that moment. Progression
of their disease was not found in their recent evaluation. Two others cases with functional
class IV and III (PSAP 86 and 57mmHg, respectively) received sodium restriction, diuretics
when necessary, losartan, digoxin and sildenafil. Three years after, the PASP estimated by
the ecochardiogram showed contradictory Results: one patient (PSAP 57mmHg) that was
functionally better (class III to I), had a higher PASP (67mmHg) on the new evaluation. One
case that presented PSAP reduction (from 86mmHg to 53mmHg, and functional class IV to
III) had performed six minutes walk test on 2007 – 430 meters and on 2010 – 537 m, showing
another functional parameter suggestive of effective treatment. Conclusions: Contrary
to idiopathic pulmonary hypertension that has a median survival time of 2,8 years, the
schistosomal disease seems to have a more indolent course, as suggested by the long time
of evolution of some cases reported. Echocardiographic estimation of PSAP seems not to be
a good evolution parameter. Schistosomal pulmonary hypertension shares similar clinical
presentation, histologic findings to idiopathic pulmonary hypertension. Consequently,
the same therapeutic options used in this group may improve the disease course of the
schistossomotic patients. Efforts must be made in order to know the natural history of
schistosomal pulmonary hypertension and better define the role of the new therapeutic
options in these individuals.
156
HEaLtH SURvEILLanCE and BaSIC HEaLtH atEntIOn, an IntEgRatIOn ExEMPLE On
tHE SCHIStOSOMIaSIS COntROL In JaBOatãO dOS gUaRaRaPES, PERnaMBUCO,
BRaSIL - 2010
francine nesello; tânia gomes de Carvalho; ana Carolina Cintra M. Medeiros; Rodriga Maria
zovka de Souza; Sérgio Carneiro vieira da Rocha; José alexandre Menezes da Silva
Universidade de Pernambuco, PE - Brasil
Introduction: Schistosomiasis is an endemic disease on the municipality of Jaboatao dos
Guararapes, which is located in the metropolitan area of Recife - Pernambuco - Brazil. The
city has approximately 700,000 inhabitants divided into 27 districts and six regions and
containing important rural area. The surveillance and control activities to schistosomiasis,
until 2008, was executed basically by municipal agents of endemic diseases with
coproscopic examinations and treatment of some positive cases. In April, 2009 it has
become a co-operation between health surveillance and the Family Health Strategy
(PSF). The proposal was to approach the surveillance and control of schistosomiasis
and guarantee medical accompanying of all positive cases to Schistosoma mansoni and
other intestinal parasites. This abstract shows the results of the integration among the
health surveillance and primary attention health between april 2009 and june 2010.
Methodology: This is a descriptive study of the surveillance, treatment and control data
from the municipality program of the schistosomiasis, among april 2009 to june 2010.
Data were obtained mainly from the Information System of the Schistosomiasis Control
Program (SISPCE) and analyzed using softwares Microsoft Excel 2007 and Epi Info windows
3.5.1. In April 2009 iniciated the process of empowerment of family health teams,
including the Community Health Agents (ACS) to raise knowledge about the disease and
identify risk factors in the community. Physicians and nurses were trained about clinical
and epidemiological aspects of the schistosomiasis. Currently, after the consultations the
patients was oriented to returned with a bring stool to survey of intestinal parasites. The
samples were periodically sent to the laboratory. The Kato-Katz was the method used in
analysis. The examinations results always return to the unit oh heath. The medication
for treatment of schistosomiasis was request to pharmaceutical assistance through the
regional heath office. The positive cases receives medical treatment and lead for specialist
consultation in case of complications. Results: Currently, 14 PSF units of endemics areas, in
two regions of the municipality are integrated with surveillance actions for schistosomiasis
control. From April, 2009 to June, 2010 were carried out 10 899 stool examinations. The
largest number of tests was conducted in 6.142 women (56%). Regardless of gender, 4.845
(44.4%) examinations were performed on people aged between 30 and 59 years. In 311
(2.9%) of the examinations were positive for Schistosoma mansoni and approximately
1.600 (15%) tested positive for other worms. Among patients positive for schistosomiasis
most 6.130 (56.3%) were females. Most of the integrated units to the proposed health
surveillance, still do not realize educational activities on prevention of schistosomiasis
in its territory and there are still difficulties feedback on patients treated or referred for
any reference specialist. Conclusions: This integration between primary care and health
surveillance showed operational advantages over the traditional approach of control of
schistosomiasis. One the main effects was in the contact with the community through
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the ACS, increased people‘s access to the clinical and laboaratorial examination. However
some difficulties remain about the informations return of the the treatment of patients.
There is also difficult to involvement of unit health staff, with health education actions on
the community. The main challenges are to maintain the integrated actions of primary
heath attention and surveillance and expanding the number of USF‘s participants.
158
an aUtOMatIC PROPOSaL fOR dIagnOSIS Of SCHIStOSOMIaSIS
andré Caetano alves firmo; Carmelo j. a. Bastos filho; Jones albuquerque; Silvana Bocanegra;
Reinaldo Souza Santos; Constança S. Barbosa
Introduction: The main purpose of this work is the development of a hardware/software
framework with low cost for the automated counting of Schistosoma mansoni eggs in fecal
examination. The system is based on the detection of eggs of S. mansoni from images of
layers of feces exam by a cascade of weak classifiers on Haar features and trained from a
particle swarm optimization algorithm in AdaBoost. Methodology: Through a partnership
among the research group Xiscanoé, the Graduate Program in Computer Engineering from
the Polytechnic School – UPE, and the Laboratory of Schistosomiasis at Department of
Parasitology CPqAM / FIOCRUZ, was used the structure and physical support of FIOCRUZ
and the blades of stool examinations for people suspected of schistosomiasis as a set of
data for this research. The implementation of this project is divided into three phases:
(First) - In this phase was analyzed the options for devices that serve as the acquisition of
images of slides of stool examinations of those infected. It was defined using a webcam
with pre-defined characteristics to ensure a better quality of images. This webcam was
connected to a microscope and captured a photo of every segment of the blade. The image
has gone through a process where filters were applied that minimized noise and effectively
shaped the best features of primitive image: contrast, brightness, saturation and hue. At
the end of this first phase, the product is an image file properly prepared for the intelligent
system can identify and count the eggs of S. mansoni. (Second) - At this stage was analyzed
the techniques of pattern recognition and elected that better fit for the resolution of the
problem. Thus, from an evaluation of the most widely used techniques, such as neural
networks, KNN and Bayesian classifiers and decision trees, decision trees was chosen as
the technique because its simplicity of implementation and less need for computational
resources. Upon election, the technique has been implemented and it was defined the
set of characteristics of the egg of S. mansoni used as the standard to be recognized by
the system. From this point, was prepared a set of tests using the resulting images of
the previous phase to calibrate the system and set minimum levels of acceptance and
tolerance of system errors. Still, as the activity of this phase, were made the first records
of performance and efficiency. (Third) - At this stage the proposal was analyzed using a
technique of computational intelligence (Particle Swarm optimization solution) to optimize
the processing time and performance of the system training. Was accomplished by
coupling the technique of intelligent computing and pattern recognition technique used
in the system. At the end of this process, the necessary adjustments were made for the
system to identify and count the eggs of S. mansoni and tested the performance of the new
system with the set of images from the first phase of this project. Results: From the sample
of 337 positive images (images that contain the egg of S. mansoni) and 202 negative images
(images that do not contain egg of S. mansoni) had the training system consisting of 10
weak classifiers forming a strong classifier capable of identifying the egg of S. mansoni. In
early tests, the system achieved an accuracy rate of 60%. The average training time of the
system was 47 seconds compared to 275 seconds without code optimization. Conclusions:
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the analysis of images acquired by the proposed system is potentially promising as can be
observed by the system performance. The low accuracy rate of the system is justified by
the small amount of sample used, where the literature suggests that the amount of sample
used must be more than 5,000 examples. The proposal has proven efficient by providing a
great relationship between training time and accuracy of the system, making it feasible to
implement the automatic solution as an aid in the diagnosis of schistosomiasis.
160
RISK faCtORS fOR SCHIStOSOMIaSIS In a LOW EndEMIC URBan aREa In BRazIL
Sandra Helena Cerrato tibiriçá; Elaine Soares Coimbra; Clarice abramo; adalberto Mittherofhe;
Milton f. Castro; Isabella O. Pinheiro; Luiz Claudio Ribeiro; Maria teresa Bustamante teixeira
Introduction: The occurrence of schistosomiasis is irregularly distributed in the state of
Minas Gerais, with areas of higher prevalence (northern part of the state, Mucuri Valley,
Belo Horizonte metropolitan region) interspersed with areas of low prevalence and others
not investigated yet. The expansion of the disease in the state occurs mainly in medium and
low prevalence areas. Most of the surveys concerning schistosomiasis investigations in Brazil
are carried out in high endemic areas. This paper aimed at investigating some demographic,
socio-economic and environmental risk factors for schistosomiasis in a low endemic area.
A cross-sectional survey was performed in the municipality of Piau, southeastern Minas
Gerais State. Methodology: The research took place in all micro-areas of Piau municipality,
of which three are urban and five are rural areas, through a cross-sectional population study
accomplished in 2007. The random population sample was defined based on drawings of
households registered by the teams of the Family Health Program (PSF), which provide health
assistance to 100% of the families. Prior to that, a study of the Basic Health Care Information
System data (SIAB) was carried out, to check its validity as a source of information for a
population-based sample size calculation. To calculate the sample size, it was estimated
a maximum prevalence of infected individuals of 5% (low endemic area). The confidence
interval was established in 95% (z = 1.96). For the regression logistics model were selected
variables with value of p, given by the chi-square test or Fisher smaller of 10% (p < 0,1)
and/or those considered theoretically important . . From 824 families enrolled in the Family
Health Program of Piau (PSF), 276 were randomly selected. A total of 910 pre-labeled dry
plastic containers was delivered for the collection of a single stool sample from each member
residing in the selected household. Each resident was assigned a unique personal identity
number. Only one stool sample of each individual was processed according to the Kato-Katz
technique . Duplicate slides were made from each stool specimen. All slides were prepared
and analyzed in collaboration with technicians from the GRS/JF. The sample examination
was made within 48 hours after collection. In addition to fecal examination, a structured
questionnaire about environmental conditions, hygiene, housing, habits, rubbish disposal
and sanitation was applied to 276 participating families. To ensure the repeatability and
reproducibility reliability and avoid bias in the interviews, a detailed and self-explanatory
manual was developed. A pilot project was conducted within 15 families prior to the wide
application of the questionnaire. Each household was geo-referenced using the GPS receiver
Garmin model II 12. The occurrence of the intermediate hosts of S. mansoni was investigated
too, in various water collections of the study municipality. The snails were collected from
lotic and lentic ecosystems in the urban and rural perimeter. These surveys were performed
in collaboration with technicians from GRS/JF. The spatial localization of the foci was
determined by instant positioning from a GPS (Global Positioning System) apparatus Results:
A total of 907 people were examined, from 276 registered families , with a loss of 10% It was
reported the first autochthonous cases of schistosomiasis in Piau, and found a prevalence of
2.0%. However, this prevalence was possibly underestimated due to the limitations of the
161
diagnostic test for areas of low prevalence. The great majority of patients were asymptomatic,
with low parasite load. The questionnaire revealed that three variables – contact with a
water body, reason for the contact and level of urbanization – were statistically related to
the risk of infection, in the final logistic regression model. Shistosomiasis was mainly found
in young adults of both sexes, and the prevalence was low in students under 14 years old,
contrasting with high endemecity areas. Three species of Biomphalaria snails were identified:
B. glabrata, B. tenagophila and B. straminea. Elimination of cercariae in any of the species
was not observed. B. glabrata, the species with greatest susceptibility of infection by S.
mansoni, predominated. Conclusions: This study identified the first autochthonous cases of
schistosomiasis in the municipality of Piau, classified as a low endemic area. We highlight the
possibility of an underestimated prevalence due to limitation of the diagnostic test for low
endemic areas. Important information were lifted concerning schistosomiasis transmission
in very low endemicity area, where school children did not appear, of the point of view
epidemic , as adults‘ infected indicators.
162
EvaLUatIOn Of SCHIStOSOMIaSIS MORBIdItY BY CLInICaL and ULtRaSOUnd
ExaMInatIOn, and gEOREfEREnCIng MaPIng tHE SEvERE CaSE In tHE EndEMIC
aREa Of ILHa daS fLORES, SERgIPE, BRazIL
Karla Carolline vieira Rollemberg; Carla virginia vieira Rollemberg; Bruno Leonardo
nascimento fernandes; anny Caroline Porto Chagas; fábio Ramalho de amorim; Marília M. B.
L. Silva; Ângela Maria da Silva; Mário adriano dos Santos; José antônio P. de almeida;amélia
Maria Ribeiro de Jesus
Universidade Federal de Sergipe, SE - Brasil
Introduction: Schistosomiasis is an important public health problem worldwide and in
Brazil. The presence of eggs of Schistosoma mansoni in periportal vessels generates
portal hypertension, a leading cause of morbidity and mortality in many patients. In the
endemic area of Ilha das Flores, Sergipe, Brazil, where we performed the present study,
the prevalence of schistosomiasis cases was 46.5% according to the PCE / SE data in 2007.
Part of the population depend on rice plantation and maintaining a close relationship
with infected snails of the genus Biomphalaria, host of S. mansoni . The objective was to
evaluate the morbidity (clinical forms, parasite load) of schistosomiasis in this endemic area
and variables associated with disease severity. Of clinical examination and ultrasonography
and socioeconomic questionnaires. Methodology: We conducted an epidemiological survey
using a questionnaire with 500 patients, 100 homes georeferenced distributed in four
villages (Bongue, Bolivar, Serrao and city center) during the period 2008 to 2010. These
patients underwent examination of stools by the methods of Kato-Katz and TF-test, interview
about socioeconomic, cultural, educational, and exposure factors, clinical examination, with
palpation of the spleen and liver and ultrasound examination to evaluate the degrees of
hepatic fibrosis and portal hipertention using Niamey criteria. This criteria considers: the
sizes of the right and left lobe liver, gallbladder, spleen, presence of ascites and collateral veins
and portal vein diameter. The geographical location of the houses was determined using a
georeferenced mapping database built in Spring version 5.0.1. Distribution of results of stool
examination were used to produce maps of prevalence and parasite loads (eggs per gram of
stool) using Crimestat ® software through the kernel estimator of risk. The maps were edited
in ArcView GIS version 9 (ESRI, Redlands, CA, USA). The analysis of the association between
infection with S. mansoni and demographic, social, educational and environmental data were
performed using SPSS version 17.0. Preliminary data of ultrasonography of 50 from the 120
patients infected with S. mansoni are shown. Results: The prevalence of schistosomiasis was
24% (120/500) BCa (Bootstrap) 95% [20.4 to 27.6], being categorized as the parasite load,
according to WHO standards, 15% (16/107) severe (> 400 eggs/g), 33% (35/107) moderate
(>100-400 eggs/g) and 52% (55/107) mild (23-100 eggs/g). The clinical presentations were:
Intestinal 66.7% (80/120), Hepatointestinal 25.8% (31/120) and hepatosplenic 7.5% (9/120).
97.4% (112/115) subjects affirm contact with natural water sources, 74.8% (86/115) were
degree III of water contact (more than 6 hours/week). We observed associations between
S. mansoni infection have any level of contact with natural water sources (PR = 1.9, 95%
CI 1.36 to 2.59, p <0.001) and drinking untreated water (PR = 6.9, 95% CI 2.44 to 19.86, p
<0.001), with an association between degree III of water contact level (> 6 hours/week) and
infection as compared with degree 0 (no water contact); RP 3.6 IC95% [1.05 to 12.32]; p =
163
0.04. Associations were also observed between S. mansoni infection and being male gender
(RP = 2.0, 95% CI 1.27 to 3.26, p = 0.003), low education level (PR = 7.1, 95% CI 2.41 to 20.93,
p <0.001), low income (RP = 1.8, 95% CI 1.28 to 2.60, p = 0.0005), being a farmer (PR = 2.7,
95% CI, 1.29 to 5.40, p <0.001) or a fisherman (PR = 3.3, CI 95% 1.39 to 7.69, p <0.001). All
patients were georeferenced in order to observe the relationship between environment and
parasite burden, clinical form that have been demonstrated by maps of risk estimates of
prevalence and parasite load. In the sonographic findings, we observed thickening of the
portal vein in only 2% (4/50) of patients, with a diameter ranging from 1.21 up to 1.49 cm.
Only 3 patients had some liver abnormalities. Conclusions: We believe this data may be an
important indicator of sanitary conditions in the living population of Ilha das Flores, Sergipe.
A high prevalence of severe forms of the disease (hepatosplenic and hepatointestinal) was
observed, although it was not confirmed by ultrasound method. In this context, identify and
treat patients, as well as understanding mechanisms that are associated with the promotion
of hepatic fibrosis becomes crucial to decrease the severe forms of the disease.
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IMPORtanCE Of PaRtnERSHIP BEtWEEn tHE EnvIROnMEntaL MOnItORIng
PROgRaM and faMILY HEaLtH In tHE COntROL Of SCHIStOSOMIaSIS In JaBOataO
dOS gUaRaRaPES, PERnaMBUCO, BRazIL.
tânia gomes de Carvalho; vitor alexandre Kessler de almeida; José alexandre Menezes da
Silva; Rodriga Maria zovka de Souza; Sérgio Carneiro vieira da Rocha; francine nesello
Secretaria de Saúde do Jaboatão dos Guararapes, PE - Brasil
Introduction: Schistosomiasis mansoni is a parasitic infectious disease transmitted by water
borne and caused by trematode Schistossoma mansoni. It is an endemic in 52 countries
worldwide. Represents a major public health problem, this endemic disease is associated with
poverty and low economic development. In the state of Pernambuco, has been historically
regarded as endemic country with high prevalence in some localities in the area of the forest.
Today the disease is also occurring in the coastal zone, in areas resulting from the disorderly
occupation in large cities. The Jaboatão dos Guararapes in the metropolitan area of Recife Pernambuco - Brazil has approximately 700 000 inhabitants, is divided into 27 districts and
geographically stratified into 06 regional constitute an endemic area for schistosomiasis.
Advocating the control of the disease, began in April 2009, a partnership between the
Environmental Monitoring Center (CTF) and the Family Health Program (PSF) in order to
bring the primary health care surveillance actions performed by agents of control of endemic
diseases , in addition to medical monitoring of patients positive for Schistosoma mansoni.
Methodology: The model of the descriptive study was used where data from surveillance
activities for the control of schistosomiasis in the period April 2009 to July 2010. One of
the activities was the training for the Community Health Agents . After training, the agents
began to distribute the jars collectors and inform the community about the importance of
performing the test for schistosomiasis. To select the area provides the transmission of the
disease, we used the hydrographic map of the city, where he was held in Surveys using method
(Kato-Katz) with population residing in the outskirts of Olho D‘água Lake (major focus of the
municipality). The results of individuals positive for schistosomiasis and other helminthiasis
are forwarded to the FHP and completion of treatment. Both teams were trained for the
new flow established. It was agreed that the Community Health Agents would make the
distribution of pots collectors and inform the community about schistosomiasis. The CTF, in
turn, received the samples and were tested parasitological (Kato-Katz). Forwarded the results
of individuals positive for schistosomiasis and other helminthiasis to perform the treatment in
the PSF. To analyze the way the partnership between CTF and PSF has been developed in the
city, showing the benefits brought to the people and the hardships faced in the period April
2009 to June 2010. Results: The partnership began with the participation of only six units of
Family Health (USF), located near the Olho D’Água Lake, all belonging to the regional in 2006.
Over time, others were added, and now make up 14 USF’s covering a wider region, including
also the regional 05. At the end of 2009, 6344 Surveys had been completed with positive
3.1% for schistosomiasis (201 positive cases) and 17.14% positivity for other helminths. For
the study period of 2010, the total number of examinations was 3114, with 110 positive
cases for schistosomiasis (3.5% positivity) and 15.66% positivity for other helminths. FHP
New Horizon was the biggest positive: 24.1%. For the entire study period, 9458 examinations
were performed, with positive 3.3%. There was a reduction of prevalence, 8.9% in April 2009
165
to 3.5% in June 2010. It was observed that the increase resulted not only in increasing the
number of monthly Surveys conducted in the municipality, but also the support of families
vulnerable to disease by Community Health Agents, the medical monitoring of positive for
schistosomiasis and conducting health education activities. Conclusions: Schistosomiasis
mansoni remains a serious public health problem, but the expansion of the partnership
between environmental monitoring and basic health care has brought benefits to the
residents of Jaboatao dos Guararapes, because there was intensification of the actions of
Schistosomiasis Control Program and the consequent increase in population assisted in
relation to this complaint, although some localities still have high positive and deserve more
attention, needing to be primarily worked. Other advantages of the partnership is that the
maps of the disease are generated quickly, with equity in the distribution of sampling, there
is a better use of human resources and structural promoting a reduction of rejection rates
and improved access to the examinations, but also the participation the Community Health
Agents, provides a better distribution of the information about the disease. As operational
difficulty has been the return of maps of positive cases treated by the PSF. The Management
of Health Surveillance intends to hold training workshops for all involved in this partnership
and continue expanding the number of USF‘s participants.
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a COMMUnItY-BaSEd PROgRaM: HEaLtH aCCESS and SCHIStOSOMIaSIS PREvEntIOn
and COntROL In JEQUItInHOnHa MUnICIPaLItY, MInaS gERaIS StatE, BRazIL
dener Carlos dos Reis; Maria flávia gazzinelli; Cristiano Lara Massara; Luiza valgas de Paula;
Helmut Kloos; Rodrigo Corrêa Oliveira; andréa gazzinelli
Escola de Enfermagem da Universidade Federal de Minas Gerais, MG - Brasil
Introduction: A community-based schistosomiasis prevention and control program was
developed following an epidemiological study in São Pedro do Jequitinhonha, Jequitinhonha
Municipality. This rural area is characterized by high schistosomiasis prevalence (39.4%), low
rates of health services utilization for diagnosis and treatment of schistosomiasis, lack of a
safe and adequate water supply system and a high level of government financial assistance
(Bolsa Família). The aim this study was to describe and assess the impact of a research-based
program for improvement of health access, community participation, and collaboration with
the Municipality Health Department to facilitate schistosomiasis prevention and control in
the study area. Methodology: Forthy-one residents of São Pedro do Jequitinhonha, including
community leaders and community members aged over 18 years and with a history of
participation in community activities and leadership were identified for participation with
the assistance of community health workers. An educational program coordinated by the
researchers was designed and implemented with the collaboration of local health professionals
and Municipality health officials. Twelve meetings were carried out with community members
and local health workers between 2008 and 2010 to improve participants‘ knowledge of the
role of public providers, researchers and community members and to strengthen community
organization and participation in the local schistosomiasis control program and to search for
possibilities for improving public health services access. Information was collected during
interviews with six community leaders on local community organization efforts and two focus
group discussions (FGDs) were carried out with the 41 community participants to elucidate
perceptions about “schistosomiasis”, “research” and “researchers” before and after program
implementation. A questionnaire was administered to the participants to better understand
the reasons for their participation in the community activities and to obtain their opinions
about major challenges and opportunities for community actions towards improving their
living conditions, local schistosomiasis prevention and control efforts and access to the
health services. All data collected were analyzed qualitatively using the qualitative content
analysis method by Bardin (Bardin 1991). Results: The adherence level of the participants
in all program activities averaged 60%, and was higher among women (83.0%). Prior to
program implementation, most participants considered both the local research program
(27.5%) and the researchers (26.2%) as a benefit to the community. Schistosomiasis was
strongly associated with lack of an adequate water supply system. After termination of the
health education activites, the majority of participants associated the high schistosomiasis
prevalence with lack of awareness of community members about disease risks. The FGDs
with community members showed also that schistosomiasis was not considered to be
a priority problem by the community. Deficiencies in sanitation conditions (81.0% of all
responses) and garbage in the streets were cited as more pressing problems the community
should address. Nevertheless, the community members and researchers recognized that
the deficient water supply and sanitation conditions needed urgent attention to reduce
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water-related diseases and implemented actions related to schistosomiasis prevention and
control. This included improvements of the existing water supply system, construction of a
public laundry/shower unit and a health education program for school children in the local
school. The community group also reactivated the community association and created a
health committee for discussing local health services issues. The participants explained their
engagement and participantion in the community group as a citizen responsibilty and a way
each resident may contribute to community improvements. Conclusions: This study reveals
that the continuing community-based program in São Pedro can promote aspirations and
efforts by this community to strengthen organizational structures and solve public health
problems related to schistosomiasis. This is in large part due to the proactive view expressed
by several community leaders and promoted by participating researchers that improvements
in participants‘ knowledge of the role of public providers, researchers, community
members and community organizations is crucial in schistosomiasis prevention and control.
Collaboration between these groups and the innovative approach they pursue are therefore
considered to play a central role in the implementation and sustainability of this program.
Key words: Community participation. Health education. Schistosomiais. Financial support:
Fundação de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG), Instituto Nacional
de Ciência e Tecnologia em Doenças Tropicais (INCT-DT), Fundação Oswaldo Cruz-FIOCRUZ,
CNPq.
168
fROM PREvEntIOn tO HEaLtH PROMOtIOn: a StUdY WItH HEaLtH agEntS Of a
MUnICIPaL SCHIStOSOMIaSIS PROgRaM
danielle grynszpan; Luciano Mendonça; fernanda Sandes Cardoso
Instituto Oswaldo Cruz, Fiocruz, RJ - Brasil
Introduction: Worldwide schistosomiasis affects more than 200 million people in 76
countries across Africa, Asia and the Americas - of these cases, a number exceeding 10
million in our country -, so that schistosomiasis remains a major health problem in Brazil.
Most outbreaks of this disease are in regions of low human development index (HDI),
which indicates low levels of education, especially if these regions remain in large areas
of irrigated land. One disseminating factor is the intense population migration towards
the Southeast region and the chaotic urban growth, along with slum problems and the
issue of sanitation. Methodology: Our study was conducted in Rio de Janeiro, the second
largest Brazilian metropolis, in which the municipal control of vectors is based on 10 areas
of oversight. Although of low endemicity, the disease is indigenous in the county, with
some episodes during the year. This paper presents the beliefs and opinions of the health
workers who deal with the schistosomiasis surveillance, since these beliefs and opinions
form the guidelines to their practices and work routines. With this main goal, there were
randomized 20 servers connected to the Coordination of Vector Control (SVC) of the
Municipal Health Secretariat of Rio de Janeiro, with an average length of service equivalent
to 19 years and variable schooling level. Semi-structured interviews were conducted, and
some questions were aimed at checking their ability to link health to socio-environmental
issues. Results: Of the respondents, 14 considered schistosomiasis a serious problem in
Rio de Janeiro because it causes much damage to the body of the people affected, while
the remainder did not have the same opinion, based on the low fatality rate of the disease
in relation to dengue. Only 11 professionals have cited the importance of combining
education with health-related measures to alleviate the schistosomiasis transmission.
Among the categories of perception that were found, the majority was linked to a vision
of health as absence of disease, that can be traced back to the 20s of last century and
does not cooperate for the perception of the dynamic health/disease. On the other hand,
the definition of health as well-being, physical, mental and social, that some professionals
have cited, comes from the 1946 World Health Organization documents, being important
to note that it has represented an important step for the incorporation of the social. Our
study has contributed as a source for the training of local health professionals, changing
the focus from the “management of many diseases” to a health promotion approach
that valued social and environmental issues. Its achievement helped the development of
other views of the realities on the part of health, including the association with the socioenvironmental. The professionals involved in the research reported that, at the beginning
of their work, they were called guards and suffered daily inspections in uniforms, as if they
were some sort of sanitary police. A point that was very criticized by the officials was the
decentralization of health,that turned the operation of the Program of Schistosomiasis
more difficult and seemed to discourage the fieldwork. Conclusions: Our data suggest the
importance of the offering of continuous formation courses and periodic workshops on
health education to help professionals to rethink their working practices and beliefs. It is
169
important that agents are able to distinguish disease prevention from health promotion,
since they are based on different paradigms - sickness or health. Health education is
presented as an integral part of health promotion, and may be a basic strategy for the
mobilization of society. We note that agents participating in this work have changed their
perceptions, linking the living conditions of communities to the health improvement.
170
171
EvaLUatIOn Of tHE InSERtIOn Of gEnEtIC PatRIMOnY Of BIOMPHAlARIA
TEnAGOPHIlA taIM (RESIStant tO SCHISTOSOMA MAnSOnI) In SnaILS COLLECtEd In
SCHIStOSOMIaSIS EndEMIC aREaS In BananaL/SP, BRazIL
daisymara Priscila de almeida Marques; florence Mara Rosa; deborah aparecida negrãoCorrêa3; Horácio Manuel Santana teles; Engels Maciel; Roberta Lima Caldeira; Áureo almeida
Oliveira; Paulo Marcos zech Coelho
Centro de Pesquisas René Rachou/Fiocruz, MG - Brasil
Introduction: In spite of the schistosomiasis control campaigns carried out in the last decades,
the transmission of the disease persists in almost all endemic areas, and new foci have been
reported. In view of this fact, alternative measures must be implemented. The transmission
cycle of the parasite depends on various factors, one of them being the presence of molluscs
genus Biomphalaria in natural water collections. Biomphalaria tenagophila is the second
more important species, being responsible for the majority of the autochthonous cases of
desease recorded in the State of São Paulo. A B. tenagophila lineage resistan to Schistosoma
mansoni has been foud 30 years ago in Taim, State of Rio Grande do Sul. This lineage has
proved to be resistant to different geographic strains of the parasite, as well as to various
burdens of miracidia. The resistance of this mollusc in due to the innate defense system
linked to the genetic aspects. The resistant character of B. tenagophila Taim is dominant, and
molecular works demonstrated that this lineage presents a molecular marker represented
by a 350pb band. This fragment is only found in B. tenagophila Taim population, and shows
a dominant character too. The use of B. tenagophila Taim as a model for schistosomiasis
biological control has been tested in some areas of Bananal/São Paulo. To identify whether
the genetic patrimony from Taim has been transmitted to the local snails are related to
detection of the molecular marker 350pb in the snails crossing descendants collected after
the introduction in the field, as well as to susceptibility tests performed for camparision of
the infection rates of the snails before and after the introduction. Previous results obteined
4 months post-introduction indicated that 36% of the young snails colleted and analyzed
in the brook Santa Inês, 43% in Lava Pés, and 3% in São José do Retiro presented the
molecular marker 350pb, wich is typical of the Taim strain, thus underlining evidence on
the estabilishment this strain in one part of the population after the introduction. After two
years, a new monitorement of the introduced areas is being performed, aiming to evaluating
the situation of the genetic patrimony of B. tenagophila Taim over time. Methodology: In
order to assess whether the genetic inheritance from Taim snails is still inserted in areas
where the introduction occurred a new collection of snails was carried out two years later
in the same brooks (Santa Inês, Lava-Pés and São José do Retiro). The collected snails were
exposed to artificial light for approximately 4 h, aiming at verifyng the natural cercarial
shedding. The speciments were then submitted to the technique of extraction, and
afterwards to PCR-RFLP for visualization of the molecular marker 350pb, which is typical
of B. tenagophila Taim lineage. Results: One hundred fifty nine snails were collected in the
brook Santa Inês, 6 specimens in Lava-Pés, and none could be foud in São José do Retiro.
All the snails examined were found negative for S. mansoni cercarie. The snails collected in
brook Lava-Pés did not present the molecular marker 350pb, but 10% of the snails collected
in the brook Santa Inês showed that typical molecular marker. It was observed that only the
172
brook Santa Inês presented the same physical characteristics, the other ones underwent
marked environmental changes. The snails examined did not present any cercarial shedding.
Susceptibility trials are being performed. Conclusions: These results showed that even two
years later, without any intervention, the genetic patrimony of B. tenagophila Taim persists
in the local snails colleted in the brook Santa Inês. The decrease in the rate of snails with
the typical molecular marker from B. tenagophila Taim, throughout a period of two years in
the same area (from 36% to 10%), thus evidencing the importance of previous molluscicide
application. The results observed concerning the other two brooks, where no snails or very
few speciments could be collected, showed the environmental changes, mainly due to
anthropic impacts, which interferences in the biology of the snail, and as a consequence in
the genetic patrimony from B. tenagophila Taim inserted in the local population. There is
a need for new introductions aiming at obtening a better understanding about this model.
Financial Support: CAPES, CNPq, PRONEX e FAPEMIG
173
anaLISYS Of tHE In vItRO POtEnCIaL SCHIStOSOMICIdaL Of tHE CandEIa ExtRaCtS
Priscila Silva grijó farani; Laura ambrósio tosta; Luisa Maria Silveira de almeida; Marcelo
Silva Silvério; ana Carolina alves de Mattos; Orlando vieira de Sousa; Paulo Marcos zech
Coelho; Eveline gomes vasconcelos; Priscila de faria Pinto
Introduction: Eremanthus erythropappus (DC) McLeisch, is a plant popularly known as
Candeia (Asteraceae), with a high prevalence in the State of Minas Gerais (SILVÉRIO, et al;
2008). The essential oil extracted from the wood of E. erythropappus has the potential to
inhibit the active penetration of the cercaria to the skin and so was considered its potential
schistosomicidal (BAKER et al, 1972). This effect was related to the presence of sesquiterpene
lactones not unsaturated. Candeia have a large therapeutic applications (SOUSA et al.; 2008)
and this specie is well distributed in the country. For this reason, we chose the ethanolic,
dichloromethane and hexane extracts of leaf of Candeia to use for to evaluate their potential
schistosomocidal in vitro experimental model. Methodology: The leaves of E. erythropappus
were collected from trees located on the campus of Universidade Federal de Juiz de Fora,
Juiz de Fora, MG, Brazil and identified by the herbarium of this university. Dried leaves
were crushed and the fragments were submitted a 80 mesh sieve. Hexane, ethanol and
dichloromethane extracts were obtained by static maceration for a period of two weeks with
six changes of solvent. After removal of solvent, and extracts were dissolved in saline plus 1%
DMSO (v/v). Swiss mice (mean weight 20 g) were infected by subcutaneous route with 100
± 10 cercariae of S. mansoni (LE strain). The Guidelines of the Ethical Committee for the use
of experimental animals of the UFJF were followed. The animals were sacrificed by cervical
fracture and perfusion was performed for collecting worms in the mesentery and liver. The
worms removed were washed in culture medium RPMI-1640 and distributed in culture
plates of six wells (four pairs per well) containing 4.0 mL of culture medium supplemented
with 5% fetal bovine serum (FBS) and 100μg/mL penicillin/streptomycin. The couples were
exposed to increasing concentrations of Candeia extracts (50, 75, 100 and 200 μg/mL). The
worms were kept in contact with the extracts for 24 hours, after that period the worms were
washed three times with culture medium and maintained in culture for more than 48 hours.
Throughout the test the worms were incubated at 37oC and 5% CO2. Analyses were performed
on Olympus inverted microscope and photographed on Canon digital camera. Results:
The experiment was conducted in two stages and the parameter used to demonstrate the
effective action of the extracts on adult worms of S. mansoni was the presence or absence of
movement and eggs in the culture medium after 24 hours of exposure to extract. As a control,
pairs of worms were kept under the same experimental conditions in absence of the plant
extracts and their behavior was observed for comparison of the parasites from the different
treatments. In the first trial were analyzed at 6 and 24 hours after addition of the extracts
at concentrations of 100 and 200 μg/mL, and observations 24 hours after the withdrawal of
the extracts. At these concentrations, all extracts (hexane, ethanol and dichloromethane)
had schistosomicidal activity. Exposed worms (males and females) presented with static,
no body movements, including the oral sucker. The damage caused by the extracts on the
adult worms has been irreversible. In the second assay analyzes were performed 24 hours
after the addition of the extracts at concentrations of 50 and 75 μg/mL. In this, only the
174
dichloromethane extract was effective, causing irreversible damage, even after removal of
the extract, the worms remained motionless and without the presence of eggs. The adult
worms exposed hexane extract showed moves very slow, 24 hours after the addition of the
extract. The ethanol extract, at these same concentrations, was not as effective on adult
worms. At both concentrations, 50 and 75 μg/mL, the worms had movement throughout
the body, however, these movements were slower than those present in control worms.
Twenty-four hours after removal of the ethanol extract, the couples of worms were again
observed and movements had more intense than in the previous analysis, but less intense
than the controls. In this observation was counted about 30 eggs from the 1st stage in the
well of 50 μg/mL subjected to ethanolic extract. Conclusions: The results obtained in the
present study confirm the activity in vitro of the extracts of Candeia on adult worm of S.
mansoni, causing total paralysis of males and females. These results, associated with low
toxicity and pharmacological effects of these extracts, are an useful tool in development of
antischistosomal new drugs. Financial Support: FAPEMIG/CPqRR/UFJF.
175
anaLYSIS Of antagOnIStIC dRUgS tO CaLCIUM CHannELS In SCHISTOSOMA
MANSONI adULt WORMS
flávia fernanda Búbula Couto; Marah Mileib vasconcelos; neusa araújo; Paulo Marcos zech
Coelho; Rafaella fortini grenfell e Queiroz;
Centro de Pesquisas René Rachou / Fiocruz, MG - Brasil
Introduction: Chemotherapy is the main alternative for reduction of schistosomiasis morbidity
in endemic areas, as well as for treatment of isolate cases of the disease. Currently, praziquantel
(PZQ) is the drug of choice for schistosomiasis treatment, being effective against the five species
of Schistosoma. Approximately, 30 years after PZQ usage the increase of resistance to treatment,
the tolerance level of the worm to the drug, as well as therapeutic failures have been observed
in several countries. There is a real and pressing need for discovering alternatives to the only
available antischistosomal drug worldwide. The complete mechanisms of action of PZQ remain
unclear, however, some effects of this drug on the parasite are well known, such as muscular
contraction, tegumental damage, and metabolic changes. Further, it has been demonstrated
that PZQ is effective due to the increase of intracellular calcium in the parasite. Calcium signaling
is fundamental for muscular contraction of Schistosoma mansoni. The control of cytosolic
calcium concentrations is also fundamental to its cell physiology as a whole, and in muscular
contraction in particular, by channels located on the tegument of the parasite. Thus, based
on the importance of this ion equilibrium for the parasite´s survival, and on the description
of channels in its tegument, the antagonists to the calcium channels are promising drugs that,
on the contrary, block the ion influx, and this can lead to a rupture of the membrane. The aim
of this work was to test in vitro different dosages of antagonists to calcium channels, Nifedipin
(NIF), Anlodipin (ANL), and Diltiazin (DIL), in S. mansoni adult worms. Methodology: Forty five
days post-infection, mice infected with 100 ± 10 S. mansoni cercariae were submitted to the
technique of retrograde perfusion of the liver, as described by Smithers & Terry (1965), using
culture medium RPMI-1640 with 8% heparin. Four worm pairs were distributed into each well of
tissue culture plates, with 6 wells (six well plates) containing 3 mL RPMI-1640 supplemented with
5% fetal bovine serum, and 100 µg/mL of antibiotics penicillin/streptomycin. Afterthat, different
dosages of NIF (0.002 , 0.001, 0.005 or 0.003 mg), or ANL (0.0005, 0.0003, 0.0001 or 0.00006
mg) or DIL (0.009, 0.005, 0.002 or 0.001 mg) were added, and maintained in an incubator at
37o C, and 5% of CO2 . Control wells, containing worms without drug addition, were also kept.
After 24 h the worms were washed with culture medium for drug removal, and maintained in an
incubator at 37o C and 5% of CO2 only in culture medium supplemented with fetal bovine serum
and antibiotics. The worms were daily observed under an inverted microscope, in order to verify
the inflicted alterations, as well as worm survival and egg-laying. Registers were made using a
camera connected to the microscope. Results: It was possible to observe that all worms exposed
to NIF, ANL, and DIL, in all the concentrations used, were dead, contracted and morphologically
changed. The control worms presented normal morphology and movements, and eggs of all
stages of development could be observed seven days after cultivation. Conclusions: Nifedipin,
Anlodipin, and Diltiazen were found to be effective in vitro killing all S. mansoni adult worms.
In this way, ex vivo and in vivo trials using drug dosages lower than DL50 are being processed in
order to assess the effects in presence of S. mansoni infection.
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angIOgEnESIS: a QUantItatIvE EvaLUatIOn In tHE LIvER Of MICE WItH
SCHIStOSOMIaSIS, BEfORE and aftER SPECIfIC CHEMOtHERaPY.
Elisângela trindade Santos.; Márcia Maria de Souza.; ana Cristina gonzalez.; zilton de araújo
andrade
Centro de Pesquisas Gonçalo Moniz, BA - Brasil
Introduction: It has been demonstrated that fibrogenesis is preceded by granulation tissue
formation (angiogenesis). Transformation of pericytes into myofibroblasts is the key event to
be then considered. In hepatic schistosomiasis, Schistosoma mansoni eggs retained within
the liver cause angiogenesis and fibrosis. Paradoxically, it has also been observed that after
curative chemotherapy, angiogenesis is also present when fibrosis undergoes regression. The
present work aims at a comparative-quantitative immuno-histochemical estimation of the
proliferated blood vessels, in the liver of mice with S. mansoni infection, before and after
curative chemotherapy. Methodology: Forty S. mansoni infected Swiss mice of both sexes
were used, 20 treated and 20 non-treated controls. Infection was by the transcutaneous
route with 50-cercarae. Sixty days following infection all the animals were treated with a
single curative dose of either praziquantel (400mg/Kg bw) or oxamniquine (100mg/Kg bw)
administered all on the same day. A surgical liver biopsy was performed immediately before
treatment, as well as 2 months afterwards. Histological, histochemical and immunofluorescent
techniques were performed for the demonstration of structural changes, laminin and α-actin.
Results: Laminin and actin were used to identify blood capillaries which were then counted.
Results obtained by morphometry revealed increased number of blood capillaries on both
groups of treated mice during fibrosis regression. Sections stained with α-actin revealed
greater number of vessels, approximately 50% more, when compared with the goup treated.
However, with laminin only a 25% difference was registered. Conclusions: The morphometric
data show that the process of regression of fibrosis in schistosomiasis is accompanied by
angiogenesis, which probably is inducing healing with remodeling of granulomas. However
the number of vessels present in the regression stage of the disease was lower in group
valued two months after treatment.
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antISCHIStOSOMaL EvaLUatIOn Of tHE IMIdazOLInE dERIvatIvE 3-(4-CHLOROBEnzYL)-5(4-nItRO-BEnzYLIdInE) IMIdazOLIdInE-2,4-dIOnE
Marek Henryque ferreira Ekert; Juliana Kelle de andrade Lemoine neves; andré de Lima
aires; tiago Moreira alves feitosa; Renata alexandre Ramos da Silva; Maria do Carmo
alves de Lima; Ivan da Rocha Pitta; Suely Lins galdino; Mônica Camelo Pessoa de azevedo
albuquerque
Introduction: Praziquantel is the drug recommended by the WHO for treatment of schistosomiasis.
Reasons for this recommendation are the good therapeutic efficacy, antiparasitic activity against all
species of Schistosoma, absence of severe collateral effects, orally administered, satisfactory costbenefits, and large clinical experience. However, this drug it is not efficient against the reinfection
and, thus, repeated treatment is a very common practice in endemic areas. This fact has resulted
in the emergence of praziquantel-resistant Schistosoma strains. In last years, a great number of
works focusing avoid this potential risk has been developed. In this sense, niridazole prototypes have
demonstrated satisfactory antiparasitic activity against Schistosoma mansoni due to the presence
of an imidazoline ring in its chemical structure. This work aimed to evaluate the in vivo antiparasitic
activity of the derivative 3-(4-chloro-benzyl)-5(4-nitro-benzylidine) imidazolidine-2,4-dione (FZ4)
against adult worms of Schistosoma mansoni. Methodology: Forty 30-day-age female mice weighing
about 30-36 g were percutaneously infected with 50 S. mansoni cercariae (BH strain). After 50 days
of infection, animals were randomly divided in four groups containing 10 mice and these animals
were individually analyzed with respect to the elimination of eggs in the stool by Kato-Katz method
(pretreatment phase). The derivative (FZ4) was prepared by mean of dispersion with a hydrophilic
polymer polyethyleneglycol (PEG). Treatment with FZ4 was realized during five consecutive days from
the 50th day after infection. Two groups were orally treated (30 and 100 mg/Kg/Day), one group was
intraperitoneally treated (70 mg/Kg/day), and last group was the control receiving only PEG. After
fifteen days of treatment, animals were sacrificed. The susceptibility of the adult worms of S. mansoni
to the studied derivative was evaluated through the recovery of worms by mean of the technique of
perfusion of the portal hepatic system and mesentery, of the ovoposition pattern by the oogram test
and the elimination of eggs in the stool by Kato-Katz method. ANOVA test was performed to verify
any differences among data from groups (α = 0.05). Results: According to the statistical analysis it
was not found significant difference (p > 0.05) with relation to the number of adult worms recovered
in the different studied groups. The average number of worms recovered in controls and in animals
treated with 30, 70 and 100 mg/Kg/day was 23.2 (± 3.3), 28.6 (± 12.5), 17.7 (± 6.9), and 27.7 (± 10.5)
respectively. There was a slightly but not significant remission of 23.6% in the number of worms in
the group of animals treated with 70 mg/Kg/day. Analysis of the intestinal fragments did not show
alteration in the ovoposition pattern, being visualized all stages of egg development with similar
percentages for all studied groups. Considering the number of eggs eliminated per gram of stool, the
employed therapeutic scheme did not reveal difference intragroup and intergroup before and after
treatment. Conclusions: In previous in vitro studies, the derivative FZ4 showed efficacy against adult
worms of S. mansoni. However, in the present work, this derivative did not show in vivo significant
activity against these worms in the drug concentrations utilized. More studies with different dosages
have been developed in attempt to better evaluate FZ4 antischistosomal activity.
178
aSSESSMEnt Of LIvER fIBROSIS In SCHIStOSOMIaSIS BY BIOCHEMICaL MaRKERS and
PLatELEt COUnt
vinícius Martins alecrim; ana virgínia Matos Sá Barreto; tibério B. Medeiros; Edmundo P. a.
Lopes; ana Lúcia Coutinho domingues; Clarice neuenschwander Lins de Morais; Sílvia Maria
Lucena Montenegro
Centro de Pesquisas Aggeu Magalhães, Fiocruz, PE - Brasil
Introduction: Schistosoma mansoni is the etiologic agent of schistosomiasis, which
affects over 200 million people around the world. In Brazil schistosomiasis is endemic in
the rural zone. Periportal fibrosis (PPF) is a characteristic of Schistosomal disease and is
the main cause of morbidity and complications that can be identified by ultrasonography
(USG). This method is the most used because it is noninvasive, simple to perform,
without the disadvantages of irradiation and be comparatively cheaper. The USG allows
the diagnosis, the degree of classification of disease, to confirm the presence or absence
of hepatomegaly, splenomegaly, periportal fibrosis and signs of portal hypertension,
indicating more severe disease. Some biological markers of fibrosis were used for the
correlation studies with liver histology and to evaluate the clinical status disease, as
the aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl
transferase (γ-GT), alkaline phosphatase (AP) and platelet count, opening an important
research field to develop noninvasive methods to assess the stage of liver fibrosis in
human schistosomiasis. Thus the aim of this work was to evaluate the association of
biochemical markers (ALT, AST, γ-GT, AP) and platelets count with the intensity of liver
fibrosis in schistosomiasis. Methodology: Forty five (45) patients with schistosomiasis
were selected at Hospital das Clinicas of Universidade Federal de Pernambuco (UFPE),
during August/2009 to July/2010. All patients were analized according to the Niamey
classification by ultrasonography examination and divided into two groups: case and
control. The case group was subdivided into two other groups (one group represented
by patients with milder fibrosis, C and D pattern; and the other group by patients with
severe fibrosis, E and F patterns). The control group was characterized by patients
without fibrosis patterns (A and B). Patients with liver diseases as hepatitis B or C, ethanol
consumption > 210 g/week in men or > 140 g/week in women and splenectomized
were excluded. Results: We observed a predominance of female patients (59%). The
predominant clinical form was the hepatosplenic (59%) and the predominant USG
pattern was severe fibrosis (E, 59%), followed by milder fibrosis (D, 22%). Among the
biochemical markers, the AP and γ-GT showed elevated serum levels proportional to
advanced stages of PPF; the platelets count were inversely proportional with severe
patterns of PPF. ALT and AST serum levels had been normal in all patterns of fibrosis.
ALT and AST serum levels differentiates the case group (severe fibrosis, E and E patterns)
as compared with control group (p = 0,03561 and p = 0,003988, respectively). AP and
γ-GT serum levels differed patients (milder fibrosis, C and D patterns) from the control
group (p = 0,001833 and p = 0,003566, respectively) and patients with severe fibrosis
(E and F pattern) as compared with control group (p = 0,0001614 and p = 0,0002975,
respectively). Only with platelets count was possible to differentiate all three groups of
patients enrolled in the study (A:B - E:F, p < 0,0001, A:B - C:D, p = 0,002693 and C:D - E:F,
179
p = 0,03916). Conclusions: Among the biological markers studied, aminotransferases
were good markers differentiating patients with no fibrosis from patients which had
severe fibrosis, FA and γ-GT serum levels showed to be good markers differentiating
patients with no fibrosis from those with fibrosis (moderate and advanced). Platelets
count showed itself as the most promising marker differentiating all of three groups of
patients. It is necessary a prospective study to confirm these data, as the formation of a
biological index for fibrosis to differentiate the degrees of PPF.
180
avaLIaR a OCORREnCIa dE vIRUS dE HEPatItES B E C EM PORtadORES dE
ESQUIStOSSOMOSE ManSOnICa EM RESIdEntES EM aREa EndEMICa, aLagOaS
Maria Sonia Correia alves; danielle Correia gama; Luciano fernandes Pereira; andrei Leite
gazzaneo; Roberta Maria Pereira albuquerque de Melo; Hugo Cabral tenório; Rozangela
Maria de almeida fernandes Wyszomirska
Universidade Federal de Alagoas, AL - Brasil
Introduction: Schistosomiasis and viral hepatitis are among the major causes of liver disease
in tropical climates, suggesting a possible association between the diseases. The aim of this
study was to evaluate the occurence of hepatitis viruses B and C in schistosomiasis-carrying
patients who live in an endemic area, Alagoas. Methodology: A coproparasitological inquiry
was made at the Municipal Laboratory of Rio Largo, through the technique of Kato-Katz.
Afterwards, there was the definition of studying groups characterized by: Group I – A group of
positive patients for Schistosoma mansoni and Group II, a control group of negative patients.
People included in the groups responded to a questionnaire with sociodemographic data
and their blood samples were collected for biochemical dosages (alkaline phosphatase,
aspartate aminotrans-ferase, alanine aminotransferase, gamma-glutamyl-transpeptidase,
total bilirubin and fractions, total protein and fractions), prothrombin time, serology for
hepatitis B (HBsAg and anti-HBc) and hepatitis C (Anti-HCV). Results: From the 3030 samples
examined, 1,702 (61.17%) were negative for helminths and protozoa; 1079 (30.61%) were
positive for helminths and 249 8.22%) positive for Schistosoma mansoni. Groups I and II
showed that bathing in the river was the main form of contact with the river, respectively,
at 64.8% and 58.2%.The sexual behavior with multiple partners was identified in 5.3% in
Group I. For the patients of this group, the mean of parasite load found was 79.1 ± 174.3.
Regarding markers for HBV (HBsAg and anti-HBc) it was identified three (1.2%) individuals
positive for HBsAg in group I and two (1.2%) in Group II (p> 0.05). For anti-HBc, 42 (17.3%)
were positive in Group I and 16 (9.8%) in Group II (p<0.05), with a high risk of contact with
viruses B for Group I.When stratified by age and gender, there was no significant risk of
contacting with virus B, significant between genera [OR=1,11; IC 95%; (0,57- 2,16)x2]. It was
observed a significant risk in relation to ages 0 -18 years [OR = 1.72, 95% CI (1.05 to 2.84)
x2] and over 18 [OR = 0.71, 95% CI (0.56 - 0.90) x2]. Furthermore, it was also observed an
association between infection by the hepatitis B virus and schistosomiasis with p<0.05 [OR
2.0 (95% CI: 1.0 - 3.72) x2]. Anti HCV was negative in both studying groups. Conclusions:
This study indicates that individuals of Group I has a medium age of 24, 92 ± 15, 28 and they
more frequently showed a profile of sexual behavior with multiple partners, suggesting that
a greater risk of contacting with anti-HBC still exists.
181
BCg ExPRESSIng SM-StOLP-2, a tEgUMEntaL PROtEIn Of SCHISTOSOMA MAnSOnI,
faILS tO PROtECt MICE agaInSt CERCaRIaL CHaLLEngE
alex Issamu Kanno; Leonardo Paiva farias; Henrique Krambeck Rofatto; Cibele aparecida
tararam; Bogar Omar araujo Montoya; Patrícia aoki Miyasato; Eliana nakano; Luciana Cezar
de Cerqueira Leite
Instituto Butantan, SP - Brasil
Introduction: The schistosomiasis vaccine field of research looks forward to develop a feasible
vaccine which achieves the same levels of protection elicited by the radiation attenuated
(RA) cercariae model. The theory is that a slow migration could expose the damaged
schistosomula to the immune system which could block further intravascular migration. The
lung is pointed as the main attrition organ in the migration process. An idea would be to
mimic the antigen presentation at the lung site using recombinant live vectors such as the
Bacillus Calmette-Guérin (BCG). This study aims at evaluating the antigen presentation of
Sm-StoLP-2 by BCG, which as a recombinant protein elicited 32% protection against cercarial
challenge in the murine model. Methodology: Strains of BCG Moreau were transformed with
expression plasmids containing Sm-StoLP-2 cDNA sequence under the mutated β-lactamase
promoter of M. fortuitum, pBlaF*. Each of three plasmids used aimed to express the antigen
either fused with the signal sequence of β-lactamase (pLA71), with the hole β-lactamase gene
(pLA73) or adjacent to a Shine-Dalgarno sequence (pMIP12), named pLA71Sto, pLA73Sto
and pMIPSto, respectively. The recombinant strains were grown in Middlebrook 7H9
medium and selected by resistance to kanamycin. Clones of each recombinant were selected
and lysed by sonication and the total protein extract analysed by SDS-PAGE and Western
blotting (WB). Using antibody previously raised in mice immunized with the recombinant
protein, following expression analysis the subcellular location of the heterologous protein
was determined by treating the total protein extract with detergent followed WB. The
antibody against Sm-StoLP-2 used in the WB was previously raised in mice immunized with
the recombinant protein. Once heterologous expression was detected by the rBCG strains,
groups of 5-wk old C57BL/6 female mice were immunized subcutaneously with 1 x 106 CFU
of the bacteria. The groups of mice immunized with rBCG-pMIPSto, rBCG-pLA71Sto, BCG or
saline were bled to evaluate humoral immune response and the spleens collected to analyze
production of cytokines by the splenocyte upon stimulation with the recombinant protein.
Thirty days after immunization a challenge was performed with one hundred cercariaes,
exposing the shaved belly of the mice for thirty minutes. After forty five days, the mice were
sacrificed and perfused to recover worms and hepatectomized to analyze egg output in the
liver. Results: The rBCG clones transformed with pMIPSto and pLA71Sto were able to express
the antigen while the pLA73Sto construct did not, when analyzed by WB. The heterologous
protein in the BCG was enriched in the hydrophilic/intracellular fraction when expressed by
both pLA71Sto and pMIPSto. The mice immunized with rBCG strains IgG levels in the serum
did not differ statistically between the groups, as measured by ELISA. The IL-4 and IFN-ɣ
cytokine producing spleen cells were determined by ELISPOT. The IL-4 production showed no
statistical difference between the groups. Animals immunized with rBCG-pMIPSto increased
production of IFN-ɣ when compared to BCG and saline groups (p<0,05), while rBCG-pLA71Sto
did not. Worm recovery after perfusion of challenged mice showed no statistical difference
182
between the groups. The same was observed for the egg counts in the liver. Conclusions:
Two recombinant BCG strains were able to express Sm-StoLP-2. Despite the presence of
signal sequence to direct exportation in the rBCG-pLA71Sto strain, the heterologous protein
was demonstrated to be enriched in the intracellular fraction of the mycobacteria in both
strains. Immunization of mice with the recombinant BCG strains expressing Sm-StoLP-2, in
spite of stimulating a specific cellular immune response was not able to induce protection
against challenge with the parasite. The understanding of the mechanisms involved in
the immunogenicity elicited by the heterologous presentation of the antigen will provide
valuable information for further research.
183
BIOLOgICaL aCtIvItY Of SIDA PIlOSA PLant ExtRaCt On SCHISTOSOMA MAnSOnI
Hermine B.Jatsa; Cíntia ap. de Jesus Pereira; Emília S. araújo; Jaílza Lima Rodrigues; vinícius
gustavo Oliveira; vanessa fernandes Rodrigues; florence M. Rosa; fernão Castro Braga;
Mauro Martins teixeira; deborah negrão-Corrêa
Universidade Federal de Minas Gerais, MG - Brasil
Introduction: According to the WHO (1997), schistosomiasis is a chronic debilitating disease
affecting more than 200 million people worldwide. The infection is highly prevalent in Africa
and in South America. For example, estimated prevalence in Cameroon and Brazil are 1.7 and
8 million, respectively. Treatment with praziquantel is mainstay of disease control. As no backups are available, there is a pressing need to develop new anti-Schistosoma drugs. Plants have
provided a number of useful clinical agents and have considerable potential as sources of new
drugs. This experimental focused on the evaluation of schistosomicidal activity of some plant
extracts that have been used for the treatment of intestinal helminthiasis by the local population
of Africa, namely, Sida pilosa (Malvaceae), Clerodendrum umbellatum (Verbenaceae) and
Ozoroa pulcherrima (Anacardiaceae). Methodology: Plants were collected in Cameroon,
authenticated by a botanist and voucher specimen deposited in the National Herbarium of
Yaoundé. Specimens were dried in the shade, powdered and the powder of S. pilosa whole plant
and C. umbellatum leaves mixed with water (100g / L) for 24 hours of maceration. The powder
of O. pulcherrima roots was mixed with water in the same proportion, and boiled for 15 min.
Solutions obtained were filtered and then lyophilized to obtain crude aqueous extracts. To test
the biological effect of each plant crude extract, Swiss male mice were infected subcutaneously
with the LE strain of S. mansoni (50 cercaria/mouse) and treated orally with the plant extract
(S. pilosa crude extract: 40 mg/kg/day; C. umbellatum leaves crude extract: 160 mg/kg/day; O.
pulcherrima roots crude extract: 40 mg/kg/day) in a daily basis from weeks 10 to 12 after the
infection. Parasite burden was accessed one week after the end of the treatment. Adult worms
recovered by perfusion were used for the in vitro evaluation of schistosomicidal activity of S.
pilosa aqueous extract and fractions (40, 20, 10, 5, 2.5 and 1.25 mg/mL for the crude extract
and 8, 4, 2, 1, 0.5 and 0.25 mg/mL for n-butanol fractions and the residue). All experiments
were carried in quadruplicate and repeated twice, using DMSO medium as negative control
group and praziquantel as positive control group. Parasites were kept for 24 h and monitored
every 12 h to evaluate mortality rate and egg production. Results: Infectivity indices showed
that oral treatment with S. pilosa extract resulted in significant reduction of worm burden and
tissue egg deposition. The results were confirmed by in vitro evaluation of S. pilosa aqueous
extract effect on isolated adult worms. Moreover, most schistosomicidal activity was present in
the n-butanol fraction. The effect was dose-dependent, whatever the fraction tested. By 24 h, all
worms exposed to 20 mg/mL of the aqueous extract and 8 mg/mLof its n-butanol fraction were
killed, whereas no deaths were observed in the control (P< 0.001). S. pilosa aqueous extract and
its fractions also inhibited egg output. At highest concentrations (20 mg/mL and 8 mg/mL), 96
% to 100 % inhibition of egg production is registered, in comparison with the negative control
group. Conclusions: Altogether, these studies show the potential of a characterized extract of
an African plant in the treatment of Schistosoma infection. Financial support: Cnpq, FAPEMIG.
184
COMPaRISOn Of a POLYMERaSE CHaIn REaCtIOn and tHE KatO-Katz tECHnIQUE
fOR tHE dIagnOSOSIS Of SCHISTOSOMA MAnSOnI In a LOW EndEMICItY aREa Of
MInaS gERaIS StatE, BRazIL
gabriel Costa de Carvalho; Letícia Helena dos Santos Marques; Luciana Inácia gomes; ana
Rabello; Luiz Cláudio Ribeiro; Sandra Helena Cerrato tibiriçá; Elaine Soares Coimbra; Clarice
abramo
Introduction: The city Piau, located in Minas Gerais state, has approximately 191 km2
and estimated 3064 inhabitants, and is part of the middle region known as Zona da Mata
Mineira. In this area there are 144 townships and seven micro-regions, all classified as
schistosomiasis low endemicity areas. The quantitative method Kato-Katz makes the
diagnosis for schistosomiasis easily, quickly and inexpensively with the visualization
of microscopic eggs in stool samples. The polymerase chain reaction (PCR) has been
developed for the diagnosis of S. mansoni, allowing a high specificity and sensitivity in
detecting DNA in stool samples from patients with low parasite load. The aim of this study
was to compare the two diagnostic methods to assess the prevalence of schistosomiasis
at a low endemicity area. Methodology: This study analyzed fecal samples from 168
inhabitants of Piau employing the techniques of Kato-Katz and PCR. The study group was
composed of 45.8% females and 54.2% male with ages ranging from 10-70 years (mean
age = 28.2 years). Informed consent was obtained from all adult participants and parents
or legal guardians of minors. The fecal samples were collected from each person and
three slides were prepared from each sample using the Kato-Katz method. From the same
samples total DNA was isolated using the QIAMP DNA Stool Mini Kit with the protocol for
DNA isolation from larger amounts of stool and PCR was performed. This work is part of a
project approved by FAPEMIG (EDT-3323/06). Results: 11 of 168 samples were positive for
S. mansoni; 7 were positive for both techniques and 4 only for PCR. The positivity rate by
the Kato-Katz method was 4.2% and by PCR was 6.5%. Comparing both techniques, X2 test
given for p value was <0.0001, indicating that the variables are associated, in other words,
Kato-Katz negative values are corresponding to PCR negative values and Kato-Katz positive
values are corresponding to PCR positive values (Kappa index = 0.766). Considering KatoKatz as diagnostic gold standard, the sensitivity of PCR was 97.7% and specificity 100%,
negative predictive value 100% and positive predictive value 63.6%. Regarding patient age,
the p value were 0.743, demonstrating that the ages are not different between positive or
negative results for schistosomiasis (n=162) as well as the variable sex (p=0.514, n=168).
Conclusions: A single PCR survey detected more cases of S. mansoni infection than three
slides of Kato-Katz stool examinations. The molecular diagnosis of parasites in Piau showed
that there is a tendency to equal outcomes between both techniques, but more studies
must exist to get acesses the divergence between diagnoses that possible can occur. It is
worth remembering that only laboratories with adequate infrastructure can perform such
procedures, especially to be avoided the risk of contamination between samples.
185
COnvEntIOnaL PCR fOR HUMan SCHIStOSOMIaSIS dIagnOSIS In StOOL SaMPLES
fROM IndIvIdUaLS WItH LOW ELIMInatIOn Of PaRaSItE EggS
teiliane Rodrigues Carneiro; Regina Helena Saramago Peralta; Bárbara de araújo Lima dutra;
Marta Cristhiany Cunha Pinheiro; Sara Menezes de Oliveira; fernando Schemelzer de Moraes
Bezerra; José Mauro Peralta
Universidade Federal do Ceará, CE - Brasil
Introduction: Schistosomiasis is still a public health problem in Brazil. The infection is widespread
in southeast and northeast, while in northern and southern endemic areas is more dispersed. The
infection’s rate and intensity within a population are influenced according to the water contact, host
immunity and professional, cultural and religious factors. The laboratorial diagnosis of schistosome
infection has been based on direct coproscopic examination and by indirect methods for detection
of antigen, antibodies and specific DNA fragments that are associated with Schistosoma mansoni
infection. The sensibility of the stool examination is variable and depends of fecal amount, the
number of eggs excreted and intrinsic factors due the loss during the procedure. A single stool
examination could impair epidemiological surveys and even individual diagnosis. The molecular
technique based on polymerase chain reaction (PCR) has been used as an alternative method for
schistosomiasis diagnostic. The main purpose of the present study was to evaluate PCR designed
for detection of schistosomiasis mansoni DNA in individuals from a low endemic area in Ceará state
Methodology: The study was conducted in the Planalto do Cajueiro, Maranguape, Ceará, Brazil. A
total of 33 stool specimens were colected for PCR evaluated considering the results of coproscopic
examination (Kato-Katz, Saline gradient and Lutz methods) and ELISA (IgG anti-S. mansoni adult
worms antigen). These samples were divided into the following groups: ELISA reactive/Others
parasites (+) = 5 samples (Group A); ELISA reactive/Others parasites (-) = 5 samples (Group
B); ELISA non reactive/ Others parasites (+) = 5 samples (Group C); ELISA non reactive/ Others
parasites (-) = 4 samples (Group D); ELISA reactive/Coproscopic examination S. mansoni (+) = 14
samples (Group E). Positive and negative DNA controls were obtained from clinical samples with S.
mansoni infection and negative samples from non-endemic area. The DNA extraction was made
as described by Santos et al. (2007). The PCR was carried out according to a protocol described
by Pontes et al.(2002) and used primers from a genomic DNA repeated sequences of S. mansoni.
Amplified products were analyzed by electrophoresis using 2.0% of agarose gel containing 0.5 μg
of ethidium bromide/mL. Results: Group A, two of the five samples were positive by PCR; group
B, 4 of 5 samples were positive by PCR and group C, all five samples were negative by PCR. Among
the four samples in group D, one was positive in PCR. This individual should be reexamined to
confirm or not S. mansoni infection. In group E, 11 of 14 samples were positive by PCR. The three
negatives samples had less than 10 eggs per gram of stool. Among the 24 individuals who showed
reactivity by ELISA, 14 samples were positive in coproscopic examination and PCR was reactive in
17 samples. Six subjects who were ELISA reactive and present stool examination negative, showed
PCR reactivity. In group E, 7 individuals were positive by KK, 9 by saline gradient and 11 by PCR,
demonstrating that ELISA sensitivity was superior when used as a single method. Conclusions: We
conclude that PCR is an important tool to improve the sensitivity in detecting S. mansoni infection
in low endemic areas. We emphasize that is very important associate the exams since none of
them has 100% of sensibility.
186
CURCUMIn-LOadEd PLga nanOPaRtICLES:
SCHIStOSOMICIdaL aCtIvItY
PREPaRatIOn
and
In
vItRO
Priscilla Paiva Luz; Lizandra g. Magalhães; Wilson R. Cunha; Kátia Jorge Ciuffi; Eduardo J.
nassar; Paulo Sergio Calefi; Emerson H. de faria; Jairo K. Bastos; vanderlei Rodrigues; Márcio
L. a. e Silva;
Universidade de Franca, SP - Brasil
Introduction: Schistosomiasis, a chronic debilitating disease caused by trematode worms of the
genus Schistosoma, affects 200 million people in tropical and subtropical regions causing more
than 280,000 deaths per year. Praziquantel (PZQ) and oxamniquine are the drugs available for
the treatment of schistosomiasis. Reduced cure rates and the failure of treatment after PZQ
administration have been reported in patients. In addition, the existence of resistant strains
reinforces the need to develop new safe and effective schistosomicidal drugs. Curcumin is the
major constituent in the rhizome of Curcuma longa (Zingiberaceae), commonly named turmeric.
Curcumin is well-known to exhibit several biological activities, including anti-inflammatory,
antioxidant, antiviral, anti-infectious and anticarcinogenic activities. In addition, curcumin possess
in vitro antiprotozoal activity against Leishmania donovani, Giardia lamblia and Trypanosoma
brucei. Recently, it has been described the schistosomicidal effect of the oil extract of C. longa
against Schistosoma mansoni infected mice. Our goal was to incorporate curcumin into PLGA
nanoparticles and to evaluate the schistosomicidal activities of the free and incorporated drug.
Methodology: The particles were prepared by the nanoprecipitation technique. In this way, the
organic phase was constituted of PLGA and curcumin dissolved in acetonitrile. The aqueous phase
was surfactant dissolved in water. The organic phase was dropped into the stirred aqueous phase at
room temperature. Acetonitrile was removed at 50ºC under reduced pressure. The final aqueous
suspension was characterized by SEM in order to evaluate the nanoparticles morphology. The in
vitro schistosomicidal activity against S. mansoni was performed against adult worms pairs. In this
way, one adult worm pair was transferred to each well of a 24-well culture plate containing 2 mL of
the RPMI 1640 medium and incubated at 37ºC in a humid atmosphere containing 5% CO2 prior to
use. At 24 h after incubation, the incorporated curcumin was added into the medium to give final
concentrations of 20, 30, 40, 50 and 100 µM. Empty PLGA nanoparticles were used as negative
control. The effects of curcumin over S. mansoni were assessed by observing the mortality rate.
Results: The obtained curcumin-loaded PLGA nanoparticles are spherically shaped. Furthermore,
the preliminary bioassay results indicate that curcumin-loaded PLGA nanoparticles possess in vitro
schistosomicidal activity against S. mansoni adult worms at 40 - 100 μM after 120h of incubation,
leading to 81% and 100% of worms death, respectively. In addition, the studies with empty
PLGA nanoparticles revealed its inactiveness against S. mansoni. Conclusions: The curcumin was
incorporated into PLGA nanopheres and exhibited promising in vitro schistosomicidal activity. In
vivo studies are in progress in order to evaluate if curcumin loaded-PLGA nanoparticles promotes
an improvement on the schistosomicidal action when compared with free curcumin.
187
dIffEREntIaL REaCtIvItY Of Igg1 and Igg4 SUBtYPES fROM tHE SCHIStOSOMIaSIS
PatIEntS agaInSt SYntHEtIC PEPtIdES, BEfORE and aftER CHEMOtHERaPY
Karen takaki flores; gabriane nascimento Porcino; Rita gabriela Pedrosa Ribeiro Mendes;
Michélia antônia do nascimento gusmão; Cristiane de Carvalho Campos; ana Carolina Ribeiro
gomes Maia; Michelle de Lima detoni; danielle gomes Marconato; Maria aparecida Juliano;
Luiz Juliano; Paulo Marcos zech Coelho; Priscila de faria Pinto; Eveline gomes vasconcelos
UNIVERSIDADE FEDERAL DE JUIZ DE FORA, MG - Brasil
Introduction: In schistosomiasis, protective immunity against infection with S. mansoni has
been related to the development of both humoral and cellular immune responses. Analyses
of antibody responses to specific antigens demonstrated high IgG4 antibody levels associated
with the slow development of immunity to S. mansoni re-infection after treatment. In
contrast, individuals who were resistant to S. mansoni infection and re-infection and living in
schistosomiasis endemic areas, produced high levels of IgG1 antibody and the results were
correlated with parasite elimination. Using potato apyrase as coating antigen in ELISA, we
demonstrated different levels of IgG isotypes against conserved epitopes shared between
this vegetable protein and S. mansoni ATP diphosphohydrolase isoforms in experimentally
infected-BALB/c mice during acute and chronic phases, after treatment and after reinfection (Mem. Inst. Oswaldo Cruz 105:374-379, 2010). Variable IgG1 and IgG4 antibody
reactivity against potato apyrase was demonstrated in patients with schistosomiasis cured
after treatment with praziquantel (Mem. Inst. Oswaldo Cruz 105:370-373, 2010). In an
attempt to detect the potential of the epitopes within synthetic peptides in prognostic of
the human schistosomiasis, these new biomolecules were tested before (BT) and after (AT)
chemotherapeutic cure with praziquantel. Methodology: Two peptides (B1LJ and B2LJ),
designed based on a conserved domain shared between potato apyrase and soluble S.
mansoni ATP diphosphohydrolase isoform, were obtained by solid-phase synthesis, and
their molecular mass and purity were confirmed by amino acid analysis and by MALDI-TOF.
The serum samples of patients with schistosomiasis (n= 26), most of whom exhibited a low
intensity of infection, were selected from a collection of samples from Penha do Cassiano, in
the state of Minas Gerais (MG), Brazil, an area endemic for S. mansoni. The patients (age range
4-83 years; 14 male and 12 female) were diagnosed by positive quantitative parasitological
stool examination by the Kato-Katz method (210 ± 77 eggs per gram of faeces). Blood was
obtained by venipuncture before and six months AT with praziquantel (40 mg/kg of body
weight for adults and children). After treatment, the patients had a clear demonstration
of chemotherapeutic cure and negative results on parasitologic examination of the faeces.
Patients co-infected with other parasites were excluded from this study. Sera samples from
healthy individuals (n= 12) were used as control. The IgG1 and IgG4 antibody reactivities
were evaluated in serum samples diluted 1:50 by ELISA (DO 492 nm), using peptides (10
µg/well) as coating antigens, peroxidase-conjugated anti-IgG1 or anti-IgG4 human-specific
immunoglobulin antibodies and OPD/H2O2 as the substrate. The results were expressed in
optical density. Results: The IgG1 antibody reactivity against B1LJ increased after treatment
(C, 0.003 ± 0.004, cutoff 0.011; BT, 0.022 ± 0.039, 9/26, 35% seropositivity; AT, 0.028 ±
0.035, 15/26, 58% seropositivity), whereas IgG4 antibody reactivity against this peptide
significantly (P < 0.05) decreased (C, 0.117 ± 0.026; cutoff 0.169; BT, 0.162 ± 0.065, 10/26,
188
39% seropositivity; AT, 0.117 ± 0.043, 3/26, 12% seropositivity). The particular analysis of
each patient after treatment revealed that IgG1 reactivity for B1LJ was maintained, increased
or reduced in one, four and two, respectively, of the nine seropositive patients, whereas
two of them were seronegative. Eight patients, who were IgG1 seronegative, demonstrated
seropositivity for B1LJ after treatment. IgG4 seropositivity for B1LJ maintained similar level
in one patient and slowly decreased in two of the ten seropositive patients, whereas another
seven seropositive patients were seronegative AT. No significant difference was observed in
the IgG1 antibody level against B2LJ before or after treatment (C, 0.021 ± 0.017, cutoff 0.055;
BT, 0.041 ± 0.046, 6/26, 23% seropositivity; AT, 0.038 ± 0.060, 4/26, 15% seropositivity).
On the other hand, the IgG4 antibody reactivity against B2LJ had significant (P < 0.001)
reduction after treatment (C, 0.109 ± 0.018, cutoff 0.146; BT, 0.170 ± 0.066; AT, 0.115 ±
0.061), decreasing also the seropositivity for this peptide (BT, 17/26, 65%; AT, 6/26, 23%). The
particular analysis of each patient revealed that six of the seventeen seropositive patients
had similar or reduced IgG4 antibody reactivity for B2LJ, whereas eleven of them were
IgG4 seronegative after treatment. Conclusions: Individual analysis revealed variable IgG1
and IgG4 reactivity against peptides before and after treatment, suggesting that 2 different
antigenic epitopes in each peptide could be involved in susceptibility and/or resistance to S.
mansoni infection. These two new biomolecules could be studied in larger populations from
S. mansoni endemic areas to test their potentiality in epidemiological studies, before and
after chemotherapy. Financial Support: FAPEMIG, CNPq, UFJF.
189
EffECt Of dExaMEtHaSOnE In tHE tREatMEnt fOR ExPERIMEntaL SCHIStOSOMIaSIS
ManSOnI
neusa araújo; Paulo Marcos zech Coelho; naftale Katz
Centro de Pesquisas René Rachou, Fiocruz, MG - Brasil
Introduction: The simultaneous use of dexamethasone or other compounds that stimulate
the metabolic enzymatic system of drugs derived from cytochrome P450, such as
antiepileptic drugs (carbamazepine, phenytoin, phenobarbital), can reduce the plasmatic
concentration of praziquantel. This study aimed at evaluating the effect of dexamethasone
and antischistosomal drugs (oxamniquine and praziquantel), in association, in the treatment
for experimental schistosomiasis mansoni. Methodology: Mice infected with 100 ± 10 S.
mansoni cercariae (LE strain) were treated, 45 days post-infection, with oxamniquine (50
mg/kg), praziquantel (200 or 400 mg/kg), dexamethasone (10 mg/kg) in monotherapy or
in association. Fifteen days after treatment, worms from mesentery and liver of the treated
animals could be collected by means of perfusion, as well as from mice of the control group
(infected and untreated animals). The liver of the animals was crushed between two glass
plates, observed under a stereomicroscope for dead worms countings. Fragments were
collected from the distal part of the small intestine of the animals, squeezed between plastic
slide and cover slip, and examined under a microscope in order to observe the oogram. The
activity indicators taken into account were: mean worm burden, worm distribution in the
mesentery and liver, worm mortality rate, and oogram changes. Results: In monotherapy,
dexamethasone (10 mg/kg) did not show either hepatic shift of worms or dead worms, and
the oogram presented S. mansoni eggs at all development stages; oxamniquine, at the dose
of 50 mg/kg, showed dead worms (41%) and oogram changes (89%); the administration of
dexamethosone and oxmniquine in association, at the same doses used in monotherapy,
caused the death of worms (45%) and oogram changes (71%). In monotherapy, praziquantel
occasioned 49% and 77% of worm mortality, as well as 60% and 100% oogram changes,
when administered at the doses of 200 or 400 mg/kg, respectively. Praziquantel (200 mg/
kg) and dexamethasone (10 mg/kg) in association caused worm death (50%) and oogram
changes (25%), and using the dose of 400 mg/kg worm mortality (60%) and oogram changes
(55%) could also be detected. Conclusions: In view of the results obtained, we can conclude
that dexamethasone on its own does not present antischistosomal activity, and when used
with oxamniquine in association does not interfere either in the worm rate in the liver and/
or in the oogram changes. The association of dexamethasone and praziquantel reduces
significantly the oogram changes.
190
EffECt Of GARCInIA BRASIlIEnSIS In InfECtIOn BY SCHISTOSOMA MAnSOnI
Liliane gonçalves vila nova; amanda Esteves Rocha; Marcos José Marques; Marcelo Henrique
dos Santos; Raquel Lopes Martins Souza
Universidade Federal de Alfenas, MG - Brasil
Introduction: To control Schistosomiasis mansoni in endemic regions, chemotherapy is the main
measure used. Nowadays, in Brazil, only two drugs are used for the treatment of schistosomiasis;
oxamniquine (OXA) and praziquantel (PRZ). They present few side effects and have a high
schistosomicidal activity, thus contributing for the infection treatment, to control morbidity and
transmission of the disease. However, OXA production was reduced because of its side effects,
such as carcinogenic and mutagenic effects. Therefore, Praziquantel is now the drug of choice for
the treatment of this disease, once it has low toxicity and wide spectrum activity when compared
to OXA. However, it has the disadvantage of being eliminated very fast by the organism and its
extensive use leads to a resistance of the Schistosoma mansoni to the drug. Therefore it is very
important to search for new therapeutic alternatives for schistosomiasis. When tested in vitro, the
active principle of Garcinia brasiliensis, epiclusianona (M-6), showed promising results against the S.
mansoni. The purpose of this study was to evaluate the effect of the extract of Garcinia brasiliensis
in Schistosoma mansoni infection in vivo. Methodology: It was used Swiss mice (weighting 20 g on
average), infected in Rene Rachou Research Center, Oswaldo Cruz Institute, with 100 ± 10 S. mansoni
cercariae (LE strain), by subcutaneous route. Forty five days post infection (pi), groups of animals
were treated with a single oral dose of epiclusianona (M-6) 50mg. The animals were euthanized
using sodium pentobarbital 3% (300 μl/mice), and perfused 15 days after the end of treatment. The
same procedures were used for the control group, comprised of infected and untreated mice. After
perfusion the recovered worms were observed with a stereomicroscope to be counted, analyzed
for activity, and classified as females, males, mated and immature. It was done digestion of liver and
intestinal tissues with the objective of quantifying eggs in these tissues. The tissue fragments were
weighed and then kept in KOH 4% solution for approximately 12 hours and then incubated at 37°
C for 1 hour for digestion. After quantifying the eggs, the results were expressed as eggs per gram
of tissue. The count of eggs was performed in triplicate. The results analyses were compared with
the control group (mice infected and untreated). Statistical analysis was performed using Anova
One Way and Turkey tests with 5% significance. Results: The average number of worms recovered
from the control group after the perfusion was 29.5, composed by 11.75 males, 9.75 females, 7.75
mated, and 0.25 immature. The group treated with M-6 the average number of worms recovered
was 21.25, composed by 8.75 males, 6.5 females, 5.25 mated, and 0.75 immature. All worms
recovered by perfusion were alive and active. The differences in the amount of worms, such as the
total amount of males, females, mated, and immature were not significant when compared to the
control group. The mean number of eggs per gram of liver was 29005, and 22037 for the control
and treated group, respectively, while in the intestine there were found an average of 21277 for the
control group, and 19650 for the treated group. However, there were no significant differences in
these results. Conclusions: This study showed that, although the extract of Garcinia brasiliensis has
been effective in experiments in vitro, we could not reproduce these results in these in vivo assays
in S. mansoni. Therefore, we can suggest that these results may have occurred due to interactions
with fluids that can both enhance as neutralize the action of the drug.
191
EvaLUatIOn Of COPROSCOPIC MEtHOdS (KatO-Katz and HELMIntEx) fOR dIagnOSIS
Of SCHIStOSOMIaSIS ManSOnI In LOW EndEMIC SEttIng In CEaRÁ StatE - BRazIL.
Marta Cristhiany Cunha Pinheiro; teiliane Rodrigues Carneiro; thaís Helena guilherme de
almeida; Sara Menezes de Oliveira; Mônica Coelho andrade; fernando Schemelzer de
Moraes Bezerra
Introduction: Schistosomiasis is a parasitic disease present in tropical regions of the Americas,
Asia and Africa, constituting a serious public health problem. The diagnostic method used by
the Schistosomiasis Control Program (PCE) for routine epidemiological surveys , is the stools
examination by Kato-Katz technique, however this is not very efficient in the diagnosis of
individuals with less than 100 eggs per gram (EPG). The relative lack of sensitivity in detecting
individuals with low parasite load is one factor that may be contributing to the maintenance
of this disease. Thus, the use of alternative diagnostic methods, which establish prevalence
rates closer to reality can contribute greatly to the success of disease control in these areas.
Therefore, our objective in this study was to perform the methods coproscopic Kato-Katz and
Helmintex ® for the diagnosis of schistosomiasis in individuals from low endemic areas in the
state of Ceará, in previously reactive serologic method ELISA, comparing the prevalence rates
coproscopic by these methods. Methodology: We initially performed by serology specific
ELISA of serum samples collected from individuals in the locality of Planalto do Cajueiro,
Maranguape-CE. Those considered as reactive been requested a stool sample, to assess the
prevalence by methods coproscopic. We conducted the following methods for parasites:
Kato - Katz Method being collected one sample per individual, and made three slides and
Helmintex® Method. All patients with eggs of S. mansoni in the feces were treated with
Praziquantel ® with 50 mg / kg / body weight in a single dose. Results: When we conduct
serology IgG- ELISA for screening of residents of the Planalto do Cajueiro, we obtained 118
(47.2%) patients reactive and 132 (52.8%) were not reactive. Of the 118 reactive patients,
only 80 bring the stools to perform the Kato-Katz technique, and these were found (9.3%)
samples positive for S. mansoni, with only one egg per slide. The first slide was a patient
found positive (14.3%), the second also a positive (14.3%) and third in five positive (71.4%).
When we evaluate the parasitic load of the population, calculated using the geometric mean
of three slides analyzed, we obtained an intensity of infection was 8 OPG, being equal to the
same intensity of individual infection in most patients, whereas all had only one egg sample.
Of the 80 samples received, only 34 had sufficient material for performing the Helmintex ®
method and found these 16 (47.1%) positive samples. For comparison between the KatoKatz and Helmintex method of the 32 samples analyzed by both , 16 were positive (50%) for
both, however, 12 (75%) of these individuals were diagnosed only by Helmintex. Thus, it was
found statistically significant differences between the Kato-Katz and Helmintex ® compared
the proportion of individuals with schistosomiasis, ie the proportion of detections by method
Helmintex ® was 46.87% while the proportion of detections by the Kato- Katz was 12.50%
(p = 0.003). Conclusions: The Helmintex method was more effective in the diagnosis of
Schistosomiasis mansoni in the study site, compared to the Kato-Katz.
192
EvaLUatIOn Of COPROSCOPIC MEtHOdS (KatO-Katz and SaLt gRadIEnt) fOR
dIagnOSIS Of SCHIStOSOMIaSIS ManSOnI In LOW EndEMIC SEttIng In CEaRÁ
StatE - BRazIL.
Marta Cristhiany Cunha Pinheiro; ana Lúcia de Paula Hanemann; teiliane Rodrigues Carneiro;
José ajax nogueira Queiroz; aparecida tiemi nagao dias; Sara Menezes de Oliveira; fernando
Schemelzer de Moraes Bezerra
Introduction: Schistosomiasis mansoni is an endemic parasitic disease and, despite the
control measures being implemented, the disease remains a major public health problem.
The Kato-Katz technique is currently recommended for diagnosis of schistosomiasis to be
quantitative, inexpensive and easy to perform. However, in low endemic areas, it is difficult
to demonstrate eggs in stool samples, mainly due to the daily variation of egg-laying or
because the diagnosis is based on a small amount of feces analyzed. Methodology: The
study was conducted at the Planalto dos Cajueiros - Maranguape-Ce, located 30 km from
Fortaleza. Was initially performed an ELISA assay for detection of IgG antibodies against adult
worm antigens of S. mansoni. Individuals considered reactive serum, were asked to deliver a
stool sample to assess the prevalence by methods coproscopic. We conducted the following
methods for parasites: Kato - Katz Method being collected one sample per individual,
and made three slides and the salt gradient method. Results: ELISA assay identified 118
individuals reactive (47.2%) and 132 non-reactive (52.8%). Of the 118 reactive, 80 provided
stool samples that by the Kato-Katz method (three slides for each sample) were found (8.75%)
positive for S. mansoni. Of the 61 stool samples analyzed by the method of salt gradient, 11
(18.0%) were positive for S. mansoni. When comparing the paired results obtained by the
Kato-Katz and salt gradient, we found that the proportion of detections of positive cases for
schistosomiasis by the method of salt gradient (19.64%) was significantly higher (P = 0.039)
than the proportion of probes by Kato-Katz (5.36%). Conclusions: The saline gradient was
more effective in detecting carriers of schistosomiasis in low endemic areas of the Kato-Katz
method.
193
EvaLUatIOn Of COPROSCOPIC MEtHOdS (SaLt gRadIEnt and HELMIntEx) fOR
dIagnOSIS Of SCHIStOSOMIaSIS ManSOnI In LOW EndEMIC SEttIng In CEaRÁ
StatE - BRazIL.
Marta Cristhiany Cunha Pinheiro; teiliane Rodrigues Carneiro; Sara Menezes de Oliveira; ana
Lúcia de Paula Hanemann; fernando Schemelzer de Moraes Bezerra
Introduction: The methods used for parasitological diagnosis of schistosomiasis vary
considerably in sensitivity, depending on the amount of feces examined, the number
of eggs excreted and intrinsic factors inherent in the loss during the procedure. In
direct methods, when the parasite load is moderate to high intensity, all techniques
coproscopic satisfactory results. However, we note that most methods currently
used have great difficulty in identifying patients with low parasite load. Thus,
the development of more sensitive for detecting eggs of Schistosoma mansoni
are desirable as methods for definitive diagnosis of this disease. In this study we
investigate two new methods coproscopic for diagnosis of Schistosomiasis mansoni in
inhabitants of an area of low endemicity. Methodology: The study was a prospective,
longitudinal, with intervention performed after the test results (parasite treatment).
This study was conducted from May/2009 to March/2010, in the town of Planalto dos
Cajueiros, located in the city of Maranguape, Northeastern State of Ceará - Brazil.
Were distributed in the homes of 118 subjects, bottles for collecting the fecal samples,
identified by name and sample number of individuals. After 24 hours, the bottles were
collected and brought to LPPBM, where the samples were aliquoted and implemented
methods: salt gradient and Helmintex ®. To perform the method of salt gradient was
followed the protocol established by Cole et al. (2009), based on the use of saline
solutions of different concentrations to form a gradient differential sedimentation
between eggs of S. mansoni and homogenized fecal or tissue. To perform the method
Helmintex ® was followed the protocol proposed by Tan et al. (2007), which through
the use of paramagnetic microspheres and exposure to a magnetic field, enables the
purification and concentration of eggs of S. mansoni, and thus a greater identification
of eggs of this parasite probably exists in the stool examined. All patients with eggs
of S. mansoni in the feces were treated with Praziquantel ® at a dose of 40 mg / kg
/ body weight in a single dose. Results: There were 80 samples and because the
amount of fecal material necessary to perform the techniques, the salt gradient was
performed in 61 samples and Helmintex ® was performed on 34 samples. Of the 61
stool samples analyzed by the method of salt gradient, 11 (18.0%) were positive for S.
mansoni, parasite load, individual ranging 02-12 OPG, whereas the parasite load of the
population was estimated at 4.1 OPG. Of the 34 stool samples examined by Helmintex
®, 16 (47.1%) were found eggs of Schistosoma mansoni. This method showed greater
variation in individual parasite load, going from 0.03 to 7.73 OPG, leaving the parasite
load in this population estimated at 0.29 OPG. By observing the results of the 32
individuals subjected to the two tested methods, we obtained 17 positive patients, 06
and 15 in the salt gradient in Helmintex ®, with 04 patients positive by both methods.
Thus, the proportion of detections of individuals with schistosomiasis by the method
194
of Helmintex ® (46.87%) was significantly higher (P = 0.022) than the proportion of
detections by the salt gradient (18.75%). We emphasize that the Helmintex ® detected
11 patients who had not been detected in the saline gradient method. Conclusions:
The method Helmintex® was more effective in detecting carriers of schistosomiasis in
low endemic areas than the method of salt gradient.
195
EvaLUatIOn Of EffECt Of PRazIQUantEL
(ECHInOStOMatIdaE: PLatYHELMIntHES)
On
ECHInOSTOMA
PARAEnSEI
Júlia Peralta gonçalves; aleksandra Menezes de Oliveira; arnaldo Maldonado Júnior; técia
Maria Ulisses de Carvalho; Wanderley de Souza
Introduction: Echinostomiasis is a food-borne, intestinal, zoonotic, snail-mediated parasitosis
caused by distomate digenean trematodes mainly of the genus Echinostoma. In the mid-1990s,
the World Health Organization (WHO) estimated that 40 million individuals were infected, and
750 million were at risk of food-borne trematodiasis infections. Echinostome paraensei Lie and
Basch, 1967 is a peristomic 37-collar-spined echinostome belonging to the “revolutum group”.
The chemotherapy of echinostomes infections may be based on mebendazole, albendazole,
praziquantel or niclosamide. Praziquantel (PZQ) is the primary treatment for human schistosomiasis,
for which it is usually effective in a single dose of 30 mg.kg-1. PZQ is a pyrazino isoquinoline
derivated and the immediate modification that can be observed in schistosomes exposed to the
drug either in vitro or in vivo is a spastic paralysis of the worm musculature. In another hand, PZQ
can induce morphological alterations in the worm tegument, initially represented by vacuolization
at the base of the tegumental syncytium and blebbing at the surface. Thus, the aim of this work
was investigated the trematocidal effect of PZQ on E. paraensei. Methodology: Hamsters were
exposed to 100 E. paraensei metacercariae. After two weeks the infected hamsters were divided in
two groups: control (not treated) and treated with single oral dose of 30.0 mg/kg PZQ. The treated
and control animals was necropsied three hours after treatment. The helminthes were recovered
and counted. The reduction of recovered of helminthes was calculated. The helminthes were
fixed in 2.5% glutaraldehyde and 4% paraformaldehyde with 0.1 M sodium cacodylate buffer. The
helminthes were washed twice in sodium cacodylate buffer, pH 7.2 at room temperature, postfixed in 1% OsO4 and 0.8% C6N6FeK4, and washed with sodium cacodylate buffer. The helminthes
were then dehydrated in an acetone series and included in EPON resin. Specimens were examined
in a JEOL 1200 Transmission electron microscope. Results: After treatment with 30.0 mg/kg PZQ
there was a reduction in recovery of adult helminthes of E. paraensei in small intestine compared
to control animals (57.4 ± 6.10 vs 0.00±0.00 helminthes), with statistically significant difference (p
<0.05, One Way ANOVA, Bonferroni post-test ). There was increased recovery of adult helminthes
of E. paraensei in large intestine compared to control animals (0.00±0.00 vs15.8 ± 3.83 helminthes),
with statistically significant difference (p <0.05, One Way ANOVA, Bonferroni post-test) after
treatment with 30.0 mg/kg PZQ. Our analyses showed in control helminthes an integral tegument
with syncytial layer after muscle layer. The cyton lies just below the muscle layer which are the
cellular organelles and the nucleus. The syncytial layer was intact in control animals. Worms treated
with 30.0 mg/kg PZQ did exhibit severe alterations on tegument surface. The syncytial layer showed
peeling and bubbles in their surface. The muscle layer was disrupted or absent. In treated animals
we had seen the severe tissue disorganization. Conclusions: Praziquantel is effective in treatment
of hamsters infected with E. paraensei with smaller doses than those used in Schistosoma mansoni.
Furthermore praziquantel promotes damage of tegument of adult E. paraensei as well in other
trematodes like Schistosoma mansoni. The damage is characterized by the presence of bubbles and
the peeling of the tegument as well as in other trematodes.
196
EvaLUatIOn Of REaL tIME PCR fOR dIagnOSIS Of SCHISTOSOMA MAnSOnI InFECTIOn
Leonardo f. da Silva; Regina Helena S. Peralta; Mariana Coimbra; Magali g.M. Barreto; Heloisa
W. Macedo; Marta guimarães Cavalcanti; José Mauro Peralta
Introduction: In the seventies, Schistosomiasis mansoni mass treatment reduced the number
of severe cases of disease and controlled morbidity in vast areas of brazillian territory.
Albeit reduction of parasite load, S. mansoni transmission was not interrupted. Individuals
resident in areas of low endemicity present low or very low intensity of infection ( < 100
eggs/g of feces) which results in increased number of negative coproscopic exams. Since
parasitological methods have decreased sensitivity when applied to samples with low egg
counts, infection prevalence is usually underestimated. Herein, we evaluate the use of real
time PCR as a complementary tool to diagnosis active S.mansoni infection. Methodology:
This study was carried out in the rural area, in the county of Sumidouro, in the State of
Rio de Janeiro, Brazil. In February and May of 2010, a total of 36 individuals of both sexes
were enrolled in this study which was approved by the HUCFF/UFRJ Ethics Comittee. After
informed consent, serum and one to three stool samples/person was collected in three
different days. For S. mansoni infection screening, two slides /stool sample were examined
by the Kato-Katz in addition to detection of serum IgG reactivity against soluble adult worm
tested by IgG-ELISA. Detection of S. mansoni DNA was performed in stool samples by a real
time PCR using probes and primers targeting the cytochrome c oxidase subunit 1 (cox1) gene
in the mitochondrial genome. Results: Stool samples analyzed by Kato-Katz were positive in
8 (22.2%) and negative in 28 (77.7%) out of 36 individuals tested. In 10 (27.7%) individuals,
S. mansoni DNA was detected in stool samples by real time PCR with ct value from 23 to 36
and in 26 individuals (72.2%) no DNA amplification was detected. IgG1 anti – adult worm
detected in serum samples was reactive in 32 (88. 9%) and undetectable in 4 (11.1%) out
of 36 individuals tested. Disagreement between parasitological and molecular diagnosis
was observed in 6 (6.3%) out of 95 stool samples that were negative in the Kato-Katz but
presented DNA amplification. In 9 (25%) individuals, IgG1 reactivity was parallel to DNA
detection in stool samples and in 3 (0.8%) individuals no immunoreativity coincided with no
DNA amplification in stool samples. Conclusions: The findings indicate that complementary
tests such as real time PCR in addition to parasitological diagnosis to detect active S. mansoni
infection augment its sensitivity and improve the diagnosis in areas of low endemicity where
low intensity of infection (<100 eggs/g/feces) is more prevalent.
197
EvaLUatIOn Of SCHIStOSOMICIdaL POtEntIaL Of EPICLUSIanOnE
aline Pereira Castro; giulliano vilela Barros; Marcelo Henrique dos Santos; Liliane gançalves
vila nova; amanda Esteves Rocha ferreira; Marcos José Marques; Raquel Lopes Martins
Souza
Introduction: The schistosomiasis is a neglected disease, which affects about 200 million people
all over the world. It is caused by platyhelminthes Schistosoma mansoni which has as habitat the
hepatic portal system of mammalian. When parasitized, the man will be able to develop or not
several clinical manifestations due the presences of larval forms (cercaria and schistosomula),
adult worm and egg. For the treatment of schistosomiasis there are some drugs that act on
different evolutionary stages of parasite and that prevent the worsening of some clinical forms
of the disease. According to World Health Organization the Praziquantel (PZQ) is the chosen
drug for the treatment of parasitized individuals, because it presents a wide spectrum of action
on S. mansoni. However, some studies describe several lineages of S. mansoni which present
resistance to Praziquantel. In this context, there is necessity of new drugs that can be used
in the schistosomiasis treatment, which will be able to reduce the incidence of this disease.
In this way, the present study evaluated the potential of Epiclusianone (M6), a substance
isolated of Garcina brasiliensis, as your potential effect schistosomicidal in the ex vivo context.
Methodology: The mice (10) were infected by 100 cercarias of S. mansoni and after 45 days of
infection were euthanized and submitted to manual perfusion in order to recuperate of adult
worms. The worms were put in culture plaque of 6 wells (4 couple per well) containing culture
medium RPMI-1640 + 5% bovine fetal serum + penicillin and streptomycin at 100µg/ml and
different concentrations of M6. The tests with the M6 were perform in three phases: in the
first phase was analyzed with 25.0; 50.0 and 100.0 µg/ml; after were made tests with 18.75;
9.375 and 4.687 µg/ml; finally tested with 16.0; 14.0 and 12.0 µg/ml; 10.0 µg/ml. Then the
worms were evaluated as the amount of worms mated, movement, contraction/shortening,
morphology, detachment of tegument and oviposition after 2, 24 and 48 hours of contact with
M6. Results: All the concentrations of M6 were effective against the adult worms of S. mansoni
in the first test phase. However, in the second test phase the concentrations of 9.375 and 4.687
µg/ml didn’t present efficiency. Finally, in the third test phase and after 2 hours of contact of
the worms with 16.0 µg/ml of M6, were observed alterations on the tegument integrity and
viability of adult worms of S. mansoni; as well as the interruption of oviposition; the absent of
mating and the shortening and contraction of the worms. The muscular contraction was one
of first effects observed in the adult worms exposed to M6, which didn’t recuperate even after
the removal of substance. For the effect of comparison the PZQ, in the concentration of 1µg/
ml according to the literature, also presents these alterations in the adult worms. However,
Oliveira et al. (2006) demonstrated that the PZQ is, also, able to inhibit the excretory activity
of adult worms of S. mansoni. However, they can recoup the normal activity, depends on drug
concentration and after the removal of this. Conclusions: In function of problems related to
the fail in the treatment of schistosomiasis with PZQ, the results obtained with this work point
to the necessity of making evaluations in vivo with the M6 and with others isolated substances
Garcina brasiliensis.
198
EvaLUatIOn Of tHE gEnOMIC dna Of SCHISTOSOMA MAnSOnI In BLOOd SaMPLES
Of MICE, BEfORE and aftER tREatMEnt WItH PRazIQUantEL
Wilma Patrícia de Oliveira Santos Bernardes; fábio Ribeiro; nilton Barnabé Rodrigues; Paulo
Marcos zech Coelho; naftale Katz; neusa araújo
Centro de Pesquisa René Rachou, Fiocruz, MG - Brasil
Introduction: Schistosomiasis mansoni is an endemic parasitic disease of worldwide
importance, and the second only to malaria in terms of socio-economic impact and public
health. Currently, there are different methodologies for diagnosis of this disease, however,
none of the known techniques shows association regarding sensitivity, specificity and
control of cure. In the present study, we propose the Polymerase Chain Reaction (PCR) for
a precocious diagnosis and cure control for schistosomiasis, using an experimental murine
model. Methodology: Female Swiss mice were infected with 50 ± 10 Schistosoma mansoni
cercarie (LE strain), by subcutaneous route. Forty-five mice were separated into three groups
of 15 animals each: the control group, with animals uninfected with S. mansoni; the untreated
infected group, and a group infected and treated with praziquantel (400 mg/kg), 45 days
post-infection, single dose, by oral route. The untreated infected mice were submitted to
perfusion 58 days post-infection, whereas the infected and treated mice were followed-up
and blood samples were collected up to 38 days post-treatment. In order to perform the
PCR, blood samples were weekly collected from the animals, using filter paper. The primers
utilized amplified a DNA sequence (121 pb) of S. mansoni. Simultaneously, two other types
of techniques for parasitological diagnosis were carried out, HPJ and eclosion of miracidia.
The worms recovered by means of perfusion were counted, and male, female and paired
worms were separately taken into account. Oogram changes were analyzed in fragments
(of ± 1 cm) from the distal part of the small intestine of mice. Oograms presenting absence
of eggs, at the initial development stages, were considered as altered. Results: Between
7-10 days post-infection, PCR presented positive results for all the animals pertaining to
both infected groups. Thirty-eight days post-treatment, PCR demonstrated that the animals
remained without activity against S. mansoni. The group of treated mice presented a mean
worm burden of 1.5 living worms per group. The untreated mice were perfused 58 days
post-infection, and all of them showed eggs and worms. Fecal samples examined by means
of the miracidium eclosion method presented a worm burden of 106, 6 released miracidia
(untreated animals), and no miracidia could be detected in the group of treated mice.
Nevertheless, when the HPJ test was used, the fecal samples examined before treatment
presented negative results for S. mansoni eggs. The untreated group showed a mean worm
burden of 15.3 related to living worms recovered after perfusion. As far as the tecidual
distribution of worms was concerned, the treated animals presented an average of 17.3% in
the mesentery, and 82.7% in the liver. In the untreated group, 61.5% of the animals showed
oogram changes, i.e., there were no eggs of the parasite at the initial development stages.
Oogram changes could not be observed in the untreated group. Conclusions: The results
obtained in the present study demonstrated that PCR is a viable alternative for diagnosis of
S. mansoni infections. PCR was able to detect precociously S. mansoni DNA, whereas the HPJ
test was not effective, even when performed 45 days post-infection, in murine model. The
miracidium eclosion method did not present satisfactory results too, since some negative
199
data were obtained when compared to the perfusion technique that showed adult worms,
as well as the oogram showing eggs at all development stages. The fact that PCR presented
positive results 38 days after treatment of the animals, led us to infer that treatment with
praziquantel (at the dose of 400 mg/kg) was not totally effective. This can be confirmed
by the reduced number of parasites or viable eggs in some treated mice. Thus, it can be
concluded that PCR has a great potential for pre-patent detection of the disease, and can
offer new approaches concerning the diagnosis for schistosomiasis.
200
EvaLUatIOn Of tHE PREvaLEnCE Of SCHIStOSOMIaSIS In tHE MUnICIPaLItIES Of
tHE HEaLtH ManagEMEnt OffICE Of aLfEnaS- Mg, BRazIL.
dener Pádua Pimenta; danilo Ribeiro ferraz; Rosângela Siqueira vieira; Liliane gonçalves
vila nova; Letícia gonçalves Resende ferreira; Rubens dos Santos vieira Júnior; Raquel Lopes
Martins Souza
Introduction: The schistosomiasis is one of the major parasitic diseases in the world and it is
known to be endemic in approximately 54 countries in American and African continents. The
etiological agent is the trematode Schistosoma mansoni which causes varied symptoms with
predominant intestinal manifestations. In Brazil the number of infected people is estimated
between 2.5 and 6 million for S. mansoni. Schistosomiasis is mostly determined by absence
of basic sanitation in peripheries of large urban centers, in natura sewage directly released
in hydric collections, and its expansion is related with population migration. The diagnosis
is usually carried by parasitological methods. Examination of faecal material by Kato Katz
(KK) technique is a widely used in epidemiological surveys. The aim of this work was to
assess prevalence of schistosomiasis and evaluate the risk of stablishing focus transmission
in the population of the cities under Health management office of Alfenas city jurisdiction.
Methodology: Accomplish the coproparasitological inquiry using the Kato-katz technique
(for the research of S. mansoni eggs of) and Lutz/HPJ (for the research of S. mansoni eggs
and others parasites) in immigrant workers employed by agribusiness companies. Parallel to
this parasitological study, it was accomplished a malacological lifting in the water collections
under jurisdiction of Health Management Office of Alfenas city. Results: During the study,
the cities Health Management office the of Alfenas-MG selected in the first phase work were
Guaranésia-MG state, Brazil e Arceburgo-MG state, Brazil. The coefficient of prevalence of
enteroparasites was of 13,5% of a total of 480 coproparasitological exams accomplished with
workers at Agrícola Monções company and dwellers of Arceburgo. In 13,5%, 32,3% were
infected with S. mansoni. The second part of this study, it was accomplished in the city of
Cabo Verde-MG state, Brazil. The coefficient of prevalence of intestinal parasites was of 56%
of a total of 214 stool sample. In 56%, 30,8% were infected with S. mansoni. During the
period of 12 months, the coefficient of general prevalence for schistosomiasis was 8,4%(58),
among 694 coproparasitological examinations. Conclusions: Infection requires contact with
freshwater where infected snails (the intermediate hosts of schistosomes) have released
cercariae larvae, infected individuals, and lack of basic sanitation. From the results of the
parasitological tests and of the encounter of snail of the gender Biomphalaria in a region
rich in dams and lakes, associated to the routine of agricultural companies of the region,
importing workers, of endemic regions from the state and country, we can conclude that
there is eminent risk of the establishment of foci of transmission in the studied region.
201
aCtIOnS PERfORMEd BY SECREtaRIa EStadUaL dE SaUdE fOR tHE SURvEILLanCE Of
SCHIStOSOMIaSIS In tHE Of StatE RIO dE JanEIRO, BRazIL.
Eduardo faraj delmas; Sergio Pereira Cunha; Cristina Maria giordano dias
CEPA /LACEN / SESDEC-RJ, RJ - Brasil
Introduction: Following the Ministério da Saúde recommendations for the decentralization
of services developed by the National Health Foundation (FUNASA), it is necessary to
strengthen the managerial structures of municipalities, aiming the planning and the
implementation of measures focused on health surveillance. In order to follow this purpose,
one of the key activity of the Malacology Laboratory of the Centro de Estudo e Pesquisas
em Antropozoonoses Máximo da Fonseca Filho (CEPA) is the training of the technical staff
of the municipalities of the Rio de Janeiro State in medical and applied malacology, aiming
the surveillance and control of schistosomiasis and other parasitic diseases transmitted by
snails. In this paper they were presented the actions performed from 2007 to 2010 as well as
the difficulties to implement the schistosomiasis surveillance in the State of Rio de Janeiro.
Methodology: Through a partnership firmed with the Laboratório de Referência Nacional em
Malacologia Médica (LRNM)/ Instituto Oswaldo Cruz/Fiocruz, courses, lectures and guided
field trips were performed for the technical staff of the municipalities of Rio de Janeiro
focusing taxonomy, epidemiology, control and distribution of lymnic molluscs, with emphasis
on Biomphalaria spp. and Lymnaea columella Say, 1817 and the African snail Achatina fulica
Bowdich, 1822. Complementary field activities were performed and the equipment needed
to malacological surveys was shown (sieves for snail capture, tweezers, containers and IPE).
The handling of GPS (Global Positioning System) was shown as an important in assigning label
biotopes. Collection techniques, packaging, tests for the presence of helminth parasites, as
well as the fixation of samples for specific identification and dissection were also presented
during the 40hs of the courses. Results: In 2007, they were trained 37 professionals from
the following municipalities: Barra Mansa (3), Barra do Pirai (4), Paracambi (7), Porciúncula
(5), Rio de Janeiro (12), Sumidouro (4) and Volta Redonda (2). In 2009, CEPA was integrated
into the structure of the Laboratório Central Noel Nutels - LACEN / RJ (CEPA / LACEN /
SESDEC-RJ) encouraging greater interaction between the districts and head responsible for
Health Surveillance, allowing the improvement of logistical support. In the same year, the
municipality of Rio de Janeiro agreed with CEPA/ LACEN a training for ten professionals of
its staff and a series of technical visits, which were performed by both technical CEPA, and
by local professionals to various parts of Alto da Boa Vista, a major locality with records of
schistosomiasis cases. Other technical visits aiming the same purpose were conducted in
the minucipalities of Macaé and Paty do Alferes. In 2010, the LRNM (FIOCRUZ) and CEPA
/ LACEN, trained 17 technicians from thirteen municipalities of the State of Rio de Janeiro
(Itaperuna: 1 professional, Angra dos Reis: 1, Rio de Janeiro: 3, Carmo 1 ; Cantagalo: 1; Campos
de Goytacazes: 1; Armação de Búzios: 2; Itaguaí: 1; Barra Mansa: 2; Trajano de Moraes: 1;
Sumidouro: 1; Mangaratiba: 1, and Mage.: 1. Conclusions: It was consensus among the
trained professionals the importance of integration of professionals, allowing debates
and approaches, aiming preventive activities against the installation of schistosomiasis, in
addition to the survey of mollusc distribution. The preparation of the mollusc distribution
map and its periodic updating, objectives of the Ministério da Saúde, only become possible
202
with the collaboration of managers and professionals in each municipality involved.
Parasitological survey of the population near the banks of streams and rivers, and illustrated
lectures addressing the health education and environmental health, should also be frequent,
especially when there are poor sanitary conditions. A major problem detected in this training
process is that very often the server does not stay in the municipality (those employees are
usually hired), and also is often dismissed during political changes, or when the server is
directed to other duties. This staff turnover demonstrates the continuing need for training
courses, lectures and technical visits for the implementation of epidemiological surveillance
and mollusk survey in the entire State, a recommendation of the Schistosomiasis Control
Program (PCE) of the Ministério da Saúde.
203
aPPROaCH On tHE tRanSMISSIOn, tHE SYMPtOMS and PROPHYLaxIS Of PaRaSItES
dISEaSES MOSt COMMOn In MaRanHãO StatE
Marjane Soares ferreira; Selma Patrícia diniz Cantanhede; andiara garcez de Souza Silva;
Hallyne davinck Mesquita Moreira; gisele Cavalcante Morais; Luciana Patrícia Lima alves;
nêuton Silva-Souza
UFPA, MA - Brasil
Introduction: Parasitic diseases are considered a major public health problems worldwide,
with the highest infection rates occur in third world countries. The forms of transmission
are intimately linked to poor housing conditions, sanitation and behavioral aspects. In
recent centuries, the battle against parasites has been conducted almost exclusively by
medical therapy, resulting in only temporary cures. For this reason, the educational projects
with the active participation of the community consist of an important tool to assist the
implementation of actions to control these diseases. In this sense, the study aimed to
conduct educational activities in underserved communities in the state of Maranhao, aimed
at promoting health. Methodology: The first stage of the study consisted of gathering
information about the main diseases caused by parasites in the state of Maranhao. From
that point, were prepared lectures on basic aspects relating to transmission, symptoms and
prevention of parasitic diseases in the selected survey. Subsequently, twelve communities
that are considered poor, were selected as target audience. Of the twelve communities
worked, seven are located in municipalities of Maranhão (Barreirinhas, Fortuna, Miranda,
São Bento, Santa Quitéria, São Domingos e Vargem Grande), while five are located in suburbs
of St. Louis, the capital state (Bacanga, Barreto, Cidade Operária, Cohatrac e Coroadinho).
The contact was established with the communities through the Health Departments of the
municipalities or representatives of civil society. In each community, were randomly selected,
20 households for administering a questionnaire in order to obtain a prior diagnosis of these
locations. For each household received a questionnaire that was answered by the owner. The
lectures were given in all communities through the slideshow, show date, and the thematic
content had explored synthesized, easy to understand language and illustrative pictures. As
a final product, information on the main parasitic diseases were summarized in the form of
educational booklet and distributed at the end of the talks. The work began in June 2008
and ending in August 2009. Results: The diseases selected from the survey were: ascariasis,
hookworm infection enterobiose, schistosomiasis, tricuriose, taeniasis, filariasis, amebiasis,
giardiasis, trichomoniasis, malaria, leishmaniasis, Chagas disease and toxoplasmosis. Of the
240 respondents, 68% said they use filtered water for drinking. However, 51% reported using
untreated water for washing fruits and vegetables, while 24% said they use filtered water
and 22% mixture of water and vinegar or water and bleach. With respect to the shares of
personal hygiene, 75% of respondents who wash their hands before meals, while 48% said
they have a habit of walking barefoot. Regarding the completion of the last stool examination,
42% reported that they do not make more than one year, 31% a year ago and 12% never did.
On the positive diagnosis of parasitic diseases, 88% responded that they have been affected
by some kind of worms. A total of lectures, recorded the participation of about 1500 people
of different ages. Conclusions: Through the observations in locus in homes visited and the
tabulation of data obtained through questionnaires, one could observe the poor quality
204
of life of the communities studied and the lack of basic information about the population
of parasitic diseases. Since the twelve communities worked, the approximate total of
participants in the talks was considered low, which warns of concerns about the motivation
of local people, with regard to knowledge about parasitic diseases. Moreover, one should
consider the importance of these participants as possible multipliers of information acquired.
Finally, we point out that social intervention through the education process constitutes an
important tool in preventing the parasitic diseases, changes in established customs and
habits of life, arousing the interest of the population as a multiplier of information and thus
promoting public health.
205
HEaLtH PROMOtIng BY SCHOOLS and COMMUnItY - SCHIStOSOMIaSIS. PERnaMBUCO
2010.
Eduardo albuquerque
Secretaria Estadual de Saude – Pernambuco, PE - Brasil
Introduction: Schistosomiasis mansoni is a parasitic disease caused by Schistosoma
mansoni and is characterized inter alia by a range of activities that go beyond the social
sector intervention and public health. The construction of an alternative Social Mobilization
and the education sector as a partner in a format different from other attempts in the
past changed the concept of the relationship between public sector partnerships in
government and non government to control the disease. Methodology: This strategy is
being used with success in different regions of Pernambuco State in control of Dengue and
is designed with the following steps: 01.Choice a school, a 02.Sensibilize (a) professor (a);
03.Professor (a) sensitizes students in the classroom, 04.Professor (a) tests of knowledge
(as) students (as) on schistosomiasis and see if they have already had the doença.Setor
Health creates conditions for students to do exams and treatment , 05 students (the) search
among family members if someone among them had schistosomiasis. Creates conditions
for family health school exams and treatment, 06 students (the) search on your street
with neighbors on schistosomiasis, Health creates conditions for neighborhood checkups
and treatment in 2006. Statistical results reported by (the) students (as) to their relatives
and neighbors. School and students receive a certificate of participation in the process of
disease control (or other award) 2007 Health re-examine the people in twelve months.
This has allowed a greater demand of health facilities for examinations and treatment.
Results: This strategy, no extra costs for the health sector education has allowed to reach
a number of other schools in the short term by facilitating the work of community health
workers providing a real disease control in some areas. Conclusions: The future of social
mobilization strategies need to overcome the profile of possible actions, establish new
concept of partnership, involving partners without burdening governmental organizations
use tools and communication strategies that cause behavioral impacts.
206
RURaL SCHOOL-BaSEd PROgRaM: SCHIStOSOMIaSIS PREvEntIOn aMOng CHILdREn
In an EdEMIC aREa In, MInaS gERaIS StatE, BRazIL.
Indira Simões Martins; Leonardo ferreira Matoso; Maria flávia gazzinelli; Cristiano Lara
Massara; Helmut Kloos; andrea gazzinelli; dener Carlos dos Reis
Introduction: School-based health education programs are considered promising for the
dissemination of public health information in areas endemic for schistosomiasis. Various
studies have shown that school health programs can increase children and adolescents’
knowledge about the importance of community participation in collective actions aimed at
endemic disease control and prevention. This need is particularly acute because children
and adolescents are considered high-risk groups for S. mansoni infection and because these
age groups can increase compliance in preventive programs. The aim of this study was to
assess the impact of a school-based program in improving knowledge about schistosomiasis
among children and adolecents in São Pedro do Jequitinhonha, Minas Gerais State, Brazil.
Methodology: The education program was carried out with the 323 resident school children
who were divided into two groups: one with children between 7-10 years old (N=256) and
the second group with adolescents 11-19 years old (N= 67). The program was carried out
by local teachers supported by two of the investigators between 2008 and 2010 and had
8 major education activities to improve children´s knowledge on schistosomiasis. They
included classroom instructions,, laboratory activities, visits to risk areas for infection such
as streams and to the local health center to increase understanding about diagnosis and
treatment of schistosomiasis. The participants responded to a pre and post-test, before and
after the educational program. The post-test included the recently developed “syllogism
test” consisting of 31 images related to infection and parasitic diseases in general. School
children were asked to choose only the images related to schistosomiasis. All data colleted
were analyzed qualitatively using the content analysis method by Bardin (1991). Answers
were divided into two categories, correct and incorrect, according to the conceptual aspect
related to the definition and transmission of schistosomiasis. In this study, only the 95
children aged 7-10 years old and the 41 adolescents aged 11-19 years old who answered
the pre and post-test and participated in all education activities were included. Results:
The results showed a high rate of incorrect answers (77.9% among small children and
82.9% among the adolescents) in the pre-test related to schistosomiasis prevention and
transmission in both school groups being. Most incorrect answers were for questions related
to schistosomiasis and river pollution with garbage. The most important schistosomiasis
related symptom was associated with the presence of white spots on the skin, mainly the
face. After termination of the health education intervention, schistosomiasis was associated
with streams containing snails and human feces. Knowledge of schistosomiasis treatment
among children and adolescents was also higher after health education. The educational
activities also contributed to the decrease of incorrect answers (56.8% in children and 56.9%
in adolescents). Six month after termination of the health education program, nearly 60% of
the participants still made correct associations in the syllogism test, with a higher rate among
the adolescent group than in the children group (p< 0.001). However, there was a large
proportion of children (26%) repeated the same wrong association between schistosomiasis
207
and river and streams pollution with garbage. Conclusions: The results showed that
school based-program for schistosomiasis with a longitudinal and interactive approach
increased the knowledge of children and adolescents about schistosomiasis prevention and
transmission, corroborating findings in other studies in northern Minas Gerais State. We
therefore recommend that innovative, long-term school-based health education program
be integrated into the curriculum of schools in Sao Pedro District with the participation of
professionals from the local health services. Key words: Schistosomiasis, School-based health
education, Rural area. Financial support: FAPEMIG, FIOCRUZ, CNPq, INCT-DT
208
SOCIaL REPRESEntatIOn Of EvERYdaY EvEntS On tHE MaRgInS Of tHE PaRdO
RIvER. IMPLICatIOnS fOR tHE HEaLtH-dISEaSE PROCESS, WItH EMPHaSIS On
SCHIStOSOMIaSIS
glêissia amorim tigre; Lúcia alves de Oliveira fraga
Universidade Vale do Rio Doce, BA - Brasil
Introduction: This paper proposes a more comprehensive and integrated study of
schistosomiasis, considering the cultural, economic and social structures of a defined group
of individuals who use the waters of the Pardo River, in Cândido Sales, Bahia for their daily
activities. Also aimed to survey the most prevalent parasitic diseases, as well as investigate
the presence of snails. Methodology: We used interviews and participant observation to
collect data and Kato-Katz method for the examinations of feces. Results: The result showed
that the daily practices of the participants are closely related to the process of transmission
of infection by S. mansoni. The occurrence and maintenance cycle of the disease indicate
the need to consider new approaches to control schistosomiasis. It was found that the
reasons why these individuals carry out activities on the river are established by objective
and subjective questions. These guys even being aware of possible infection with S. mansoni
still go ahead with their activities. Of the three locations studied the highest percentage
of infection occurred in an area known as Sucuiú River (42.3%). Regarding the distribution
of infection by gender was found that there is a homogeneous distribution in both sexes.
Conclusions: Although the Pardo River is considered a vehicle of disease, it still accounts
for participants, synonymous with wealth, source of life, healing, fun, pleasure and income.
It is very important knowledge about the practices and their implications in the healthdisease, which can direct local managers to plan and operationalize measures to control
schistosomiasis.
209
COMPaRatIvE StUdY Of tHE tUBERCLES Of MaLE SCHISTOSOMA MAnSOnI USIng
COnfOCaL LaSER and SCannIng ELECtROn MICROSCOPY
Roberto Carlos ferreira João; Cléria C. Mello-Silva; viviane da Silva Costa; Cláudia Portes
Santos;
Instituto Oswaldo Cruz – LAPSA, RJ - Brasil
Introduction: Schistosoma mansoni Sambon, 1907 was first examined using electron
microscopy in 1955 and, due to its medical importance, is still investigated in this way. A major
target for study is the worm’s tegument, where adaptive changes have occurred in response
to the rapid transition from snail to human host and to the transmission mechanisms across
it which enable help the parasite’s survival within the host. The present work includes a
comparative study of the dorsal tubercles of the male S. mansoni using confocal laser
scanning microscope (CLSM) and scanning electron microscopy (SEM). Methodology:
Parasites were recovered by perfusion of the hepatic portal blood vessels of infected mice.
For SEM studies, the worms were fixed in 2.5% glutaraldehyde in 0.1 M cacodylate buffer for
30 min and rinsed in the same buffer. After post-fixation in 1% OsO4 for 30 min, specimens
were dehydrated in a graded acetone series and dried using the critical point method with
CO2. The specimens were mounted using cellotape on aluminium stubs and coated with a 20
nm-thick layer of gold. The samples were examined in a Zeiss 940 DSM microscope. For CLSM
studies, specimens were fixed in 4% paraformaldehyde (PFA) in 0.1 M PBS for 5h, transferred
to TRITON X-100 0.5% in PBS and washed in PBS. They were then incubated overnight with
faloidin conjugated to FITC 1:700 in PBS and mounted on semi-permanent slides in PBS with
2.5% de 1,4-diazabiciclo-(2,2,2)-octano-trietilenodiamina and 50% glycerol at pH 7.2. Studies
were performed using an Olympus BX51 with Fluoview version 3.2. Results: The tegument
on the dorsal surface of the male S. mansoni is known to be covered with tubercles. A general
view of the tubercles indicates that they have an almost uniform shape but an irregular
distribution. Each tubercle is composed of a group of spines which appear to arise from the
muscular fibers of the tegument, forming dome-shaped structures of a similar height. Some
of these have pores through which sensillae emerge. Using the faloidin technique, it can
be seen that the tubercles appear between the circular muscle fibers with the longitudinal
fibers passing underneath. The tubercles are either disposed in pairs or separate, and, in
the area between them, the muscular fibers of the tegument are interrupted by small holes
in which sensorial papillae can be seen. The confocal images show that within these holes
small spines occur in a deeper layer, suggesting that they may represent incipient tubercles.
Conclusions: It is clear from these results that the tubercles found on the dorsal tegument of
male S. mansoni require further study.
210
EaRLY PROdUCtIOn Of REaCtIvE OxYgEn SPECIES (ROS) BY BIOMPHAlARIA
TEnAGOPHIlA HEMOCYtES fROM tHE taIM StRaIn, WHICH IS RESIStant tO tHE
InfECtIOn WItH SCHISTOSOMA MAnSOnI
Cintia ap. de Jesus Pereira; Emília Souza araújo; Leonardo Rodrigues; Luciano S. a. Capettini;
Michelle C. de Rezende; florence M. Rosa; vanessa fernandes Rodrigues; Paulo Marcos zech
Coelho; ary Corrêa Junior; deborah negrão-Corrêa
Introduction: Most studies show that hemocytes are the main elements involved in the
destruction of Schistosoma mansoni sporocysts present in snails of the genus Biomphalaria,
intermediate hosts of this trematode. Thus, the ability of activating hemocytes during the
infection with the parasite may define its survival inside the invertebrate host. One of the
possible mechanisms involved in the death of the parasite depends on the production of
reactive oxygen species (ROS) by activated hemocytes, which may injury the integument
of S. mansoni sporocysts. The purpose of this experimental work was to comparatively
evaluate the ROS production by hemocytes of B. glabrata from the BH strain (which is
highly susceptible to infection with S. mansoni), of B. tenagophila from the Cabo Frio strain
(partially susceptible to the parasite) and of B. tenagophila from the Taim strain (resistant to
the parasite) during the experimental infection with S. mansoni. Methodology: The volume
of ROS production was determined in circulating hemocytes obtained from non-infected
snails or after 12 h and 24 h of infection with 20 S. mansoni miracidia in different strains
and species tested. Hemocytes were restimulated in vitro with total antigen of axenicallytransformed primary sporocysts (Ag/esp) or with Zymosan in the presence of a fluorescent
marker – 2’,7’- -Dichlorofluorescein diacetate (DCFCH-DA). In the presence of oxygen
derivatives, a DCFCH-DA oxidation takes place yielding a fluorescent product which was
measured in a fluorometer using 485nm wavelength of excitation and 530 nm for emission.
Results: have shown that hemocytes recovered from different species of snails infected
for 12 h or 24 h and restimulated for 60 min with Ag/esp produce ROS in similar amounts.
Once activated by the infection, these cells also respond to zymosan stimuli. However, some
non-infected hemocytes recovered from B. tenagophila from the Taim strain have produced
increased amounts of ROS after Ag/esp stimulation, up to 5 times larger than that produced
by hemocytes from susceptible and non-infected strains. Conclusions: Data suggest that an
early activation and production of ROS by circulating B. tenagophila heocytes from the Taim
strain may play a role in this strain’s resistance to the natural infection with S. mansoni.
Financial support: FAPEMIG, Capes and Pronex.
211
EvaLUatIOn MOLLUSCICIdE CHAMAESYCE HIRTA MILL SP. fOR BIOMPHAlARIA
GlABRATA
Luciana Patricia Lima alves; Marta Martins almeida; Selma Patricia diniz Cantanhede;
Joedilsa Sena; Hallyne davinck Mesquita Moreira; andiara garcez de Souza Silva; Marjane
Soares ferreira; nêuton Silva-Souza; Jackson Ronie Sá da Silva; alessandra Leda va
Universidade Estadual do Maranhão, MA - Brasil
Introduction: Schistosomiasis mansoni is a parasitic infection caused by the trematode
Schistosoma mansoni, which has as its intermediate host snails of the genus Biomphalaria.
Among the species of this genus, Biomphalaria glabrata is more important in transmitting
this disease because it has a rate of 70% natural infection and high susceptibility to various
strains of the parasite. One of the measures to reduce transmission of schistosomiasis is
intestinal mollusc population control through the use of molluscicides. Due to a number
of disadvantages with respect to the use of synthetic molluscicides (production expensive,
low selectivity and harmful to the environment), many plants have been investigated for
their potential molluscicide. Chamaesyce hirta Mill sp the species is also reported as to its
molluscicidal activity for the snail Lymnaea acuminata. In this context, the present study,
the molluscicidal activity of crude ethanol extract of the leaves and the plant (whole) of
the species C. hirta Mill sp. in snails B. glabrata, from the municipalities of São Luís and São
Bento (MA). Methodology: Samples of the plant species were collected in neighborhood
São Cristovão (São Luís city) and identified by botanical MSc. Ana Maria Maciel Leite. The
identification of specimens of C. hirta Mill sp was based on morphological characteristics.
The leaves of C. hirta Mill sp and plants (whole) were stored in glass containers with absolute
alcohol for the extraction plant for a period of two weeks. After this period, the materials
were filtered and evaporated under reduced pressure in rotovap until the complete removal
of the solvent. The extracts were stored in a freezer at -20 ° C. Snails B. glabrata were
collected from natural breeding sites located in Sao Bento (neighborhood Outra Banda) and
São Luís (neighborhood Coroadinho). Data collection was performed using metal clamps.
The natural breeding sites of B. glabrata were georeferenced with the aid of a portable GPS,
and their geographical coordinates for São Luís and São Bento: S 02 º 33 ‘40.1‘‘and HO 44
º 16‘ 00.6‘‘; S 02 º 41 ‘42.1‘ ‘HO and 44 º 49‘ 24.1‘‘. The snails collected were transported
to the Parasitology Laboratory of Human UEMA and kept under controlled conditions
of maintenance. Part of snails collected was separated for the confirmation of species,
which was determined according to morphological and conchologically parameters based
on literature. To check the positivity, the snails were exposed to light and heat three 60W
bulbs for an hour once a week, over a period of forty days. These snails were examined in a
stereomicroscope and positivity was indicated by the presence of cercariae of S. mansoni.
For evaluation of molluscicidal activity were used snails of São Bento and São Luís negative
for S. mansoni, proceeding according to the methodology of WHO (1965). Results: In the
evaluation of molluscicidal activity of crude ethanolic extract of leaves of C. hirta in snails of
São Bento, it was found that some were above the surface of the extractive solution. Such
behavior does not represent a defense mechanism to escape from the solution, since these
animals are usually characteristic in this laboratory, when placed in aquariums. During the
experiment with this statement, only two snails died. In contrast, the test of the ethanol
212
crude extract of the plant (whole), no specimen died. With regard to experiment with snails
of São Luís, was tested first ethanol crude extract of the plant (whole). At 72 and 96 hours of
the experiment, four and three specimens B. glabrata died, respectively. However, this result
is irrelevant, since, for the same time intervals were no deaths in control group B. glabrata.
In assessments of the extracts under study, it was observed that the dead snails retracted
cephalopodal the mass into the shell, without release of hemolymph. The literature reports
that the molluscicide in snail can promote two mechanisms that explain his death: retraction
of cephalopods into the shell, releasing hemolymph or abnormal extension cephalopod out
of the shell. With regard to mobility and status of the body cephalopodal clam survivors
during the experiments with the extractive solution, no change was noted. As for eating, all
surviving snails to the action of ethanol extracts of normally fed fresh lettuce. Conclusions:
Through the results, we conclude that the plant C. hirta was inactive for molluscicidal B.
glabrata from São Luís and São Luís at a concentration of 100 ppm, because the amount of
clams killed by the action of extractive solution is minimal compared to that required by the
WHO (1983).
213
EvaLUatIOn Of natURaL SCHISTOSOMA MAnSOnI InfECtIOn Of BIOMPHAlARIA
STRAMInEA In tHE PaRQUE EStadUaL dO SUMIdOURO, LagOa Santa, MInaS gERaIS,
BRazIL
amanda Cardoso de Oliveira Silveira; Érica Oliveira dias; Mônica Johanna Lukestik; Sabrina
Lúcia Marçal dos Santos; Luiz alberto dolabela falcão; Liana Konovaloff Jannotti-Passos
Introduction: Schistosomiasis is a parasite chronic disease caused by the helminth of genus
Schistosoma. The infection is common in parts of Africa, South America, Middle East and
Asia where it is a leading cause of morbidity and mortality. Recent estimates suggest that
700 million people are at risk worldwide with almost 200 million infected only in Africa.
The digenetic blood fluke, Schistosoma mansoni, is one of the major causative agents. The
basic life cycle of the parasite alternates between generations, with a sexual stage in the
definitive vertebrate host (man) and an asexual stage in a molluscan host (Biomphalaria).
A relatively short-lived, the miracidia, hatches from the egg found in human stool and
infects the snail, where it gives rise to sporocysts that produce numerous free-swimming
cercariae. In Brazil, there are 11 species and 1 subspecies of Biomphalaria, only B. glabrata,
B. straminea and B. tenagophila are found naturally infected with S. mansoni. Although
there is a large knowledge about the presence and distribution of Biomphalaria in different
habitats in Brazil, there are few studies in areas of environmental preservation. The Parque
Estadual do Sumidouro (PES) is an Environmental Preservation Area - EPA, in Lagoa Santa city,
representing a region composed of temporary ponds of reference in the karst landscape. The
aim of this study was to evaluate of natural S. mansoni infection of B. straminea in the PES
through a malacological survey preliminary. Methodology: There were four visits to the lake
of PES, the first for demarcation and georeferencing of collection sites and the others for
the effective collection of snails, carried out between March and June 2010. The snails were
collected with one standard adapted metal scoop, which was introduced on average five
times in each determinate point and all live snails larger than 2 mm diameter were washed
and separated into vials properly cataloged with a capture card, containing the date and
collection site. The identification morphologic and exam of snails by exposure to cercariae
S. mansoni were performed in the Moluscário at the Centro de Pesquisas René RachouFiocruz, Belo Horizonte. Results: A total of 339 snails were collected in the study area being
223 B. straminea (65,78%), beyond Pomacea sp. (13,27%) and Melanídeos (20,94%). No
B. straminea examined were infected with S. mansoni. Conclusions: This study provides
preliminary evidence of enabling environment for the development of various specimens
of snails. Although no specimen positive snails of B.straminea was captured in the park,
the large quantities of this specie indicates possible risk of S. mansoni infection. So it’s
recommended to carry out periodic monitoring of the site.
214
EvaLUatIOn Of tHE SPECIES and SPaCE dIStRIBUtIOn Of BIOMPHAlARIA SP. In tHE
MUnICIPaLItY Of ILHa daS fLORES, SERgIPE, BRazIL
delmany Moitinho Barboza; verônica de Loudes Sierpe Jeraldo; Claudia Moura de Melo;
Mario adriano dos Santos; nataly Cardoso Santos; Cangjie zhang; guilherme Loureiro
Werneck; José antônio Pacheco de almeida; amélia Ribeiro de Jesus
Universidade Federal de Sergipe / Hospital Universitário, SE - Brasil
Introduction: Schistosomiais is a parasitic disease caused by the digenetic trematode,
occurring in 74 countries, affecting approximately 200 million people. Schistossoma
mansoni is the species found in the Americas, and is a serious health problem in
Brazil, where it takes place in 19 states. The state of Sergipe presents the second
highest prevalence of the country (11.77% in 2007). The expansion of the disease is
connected with the geographical distribution of the water snail intermediary host and
its compatibility with the different parasite species, therefore, studies wrapping SIG and
schistosomiais becomes essential to understand the maintenance of the high rates of
prevalence and maintenance of ways of transmission of this disease. Ilha das Flores is
located in the north part of the state of Sergipe, on the margins of the San Francisco
river, the main river from the Northeast Brazil, and largely used for irrigation purposes.
Data from the Schistosomiasis Control Program (PCE) of the Health Official of the State of
Sergipe, collected in 2005 and 2007 showed a high prevalence for schistosomiais (46,45
%) in this municipality. According to data from IBGE raised in 2007, Ilha das Flores has
8.598 inhabitants, 85,35 % of those has running water, but with frequent failures of
pipe line water, and only 3,30 % of the urban population has a sanitation system. The
main economic activities of this municipality are agriculture and fishing activities. Rice
culture is the main product which requires an almost continuous water environment.
The municipality has an extensive irrigation system that was built by CODEVASF
Methodology: Georeferenced airborne photo and satellite images of Ilha das Flores was
obtained. An acquisition of control coordinates were acquired in the field with GPS. In
the cartographical base restored from the above-mentioned air photographs we mapped
the urban area containing houses and streets of the municipality and the villages, the
natural water sources, and the representative use of the land of principal thematic classes
(rice field, pastures, waste grounds, and others). Taking into account the biggest possible
covering of the area where there were cases of schistosomiais, nine collection spots were
selected for water snails. These nine water sources were identified in the field through
GPS, and from August 2009 to July 2010 a monthly screening was carried out in search
of snails and those captured were sent to the Laboratory for calculation of the density,
classification and determination of infection by S. mansoni. The identification of the
types was carried out by dissecation of the shellfish and morphological evaluation of the
shell. The infection by S. mansoni was carried out through the classic technique of light
exposure. The results of the malacologic study were put into a geo-referenced database
and a subsequent filling out of space consultations and generation of maps showing the
characteristics of the arboreta of Biomphalaria. The geo-processing information was
carried out on the SPRING platform 5.0 Results: A total of 13,873 snails were collected
between September 2009 and July 2010, being 9,799 (70.6%) examples of B. glabrata in
215
five different water collections, and 4,074 (29.4%) of B. straminea, in four water sources.
Of the total of B. glabrata captured, 113 (1.1%) was infected by S. mansoni and of the B.
straminea captured, 42 (1,0 %) was infected. At the collection spot 09 2,817 B. glabrata
(20.3%) were found and 15 (0.5%) was infected by S. mansoni, representing the lowest tax
of infection found in the study. At the collection spot 1, B. stramínea was identified and
we found the highest tax of infection (8.8%). The highest tax of infection for S. mansoni in
B. glabrata was 1.8 % at the collection spot 5 Conclusions: B. glabrata and B. straminea
were found in the majority of the studied spots and are associated with the transmission
of schistosomiais in the endemic area of Ilha das Flores, but the population of B. glabrata
was higher than B. straminea. Such a fact along with studies carried out in the decade
of 80 which showed a higher frequency of B. straminea in the region of the low São
Francisco river (regions around Ilha das Flores), suggests that these two species are
interacting in the process of competitive dislocation. We also saw that some natural water
sources used by the population for leisure, occupational practices (cultivation of rice) and
domestic activities serve as arboreta for Biomphalaria sp. infected by S. mansoni, and
the populations of shellfish in this area did not suffer major influence of rain, due to the
culture of rice plantation which requires regular irrigation
216
ExPERIMEntaL InfECtIOn Of SUBSPECIES BIOMPHAlARIA TEnAGOPHIlA GuAIBEnSIS
(MOLLUSCa: PLanORBIdaE) WItH SCHISTOSOMA MAnSOnI SaMBOM, 1907
Lidiane Bento Braga; Liana Konovaloff Jannotti-Passos; Omar dos Santos Carvalho; Roberta
Lima Caldeira
Instituto René Rachou –Fiocruz, MG - Brasil
Introduction: Among the Biomphalaria species that occur in Brazil three are regarded as
intermediate hosts of Schistosoma mansoni, B. glabrata, B. tenagophila and B. straminea.
Investigations on experimental infection using B. peregrina B. cousini and B. amazonica
have shown that they are potential hosts for the trematode. A subspecies of planorbid
snail, B. tenagophila guaibensis was described for Paraense in 1984. It has been found,
until now, along the coastal belt of the Brazilian state of Rio Grande do Sul (RS) and the
middle part of Uruguay. This was classified a subspecies for similarities with B. tenagophila
in the aspect of the shell and the others organs of the genital system in especially due
to presence of the vaginal pouch. Considering the facts that little is known about B. t.
guaibensis and that the more similar snail, B. tenagophila, is susceptible for S. mansoni the
aim of the present study was to assess the susceptibility of the subspecies to S. mansoni
infection, either as intermediate hosts and/or as potential hosts. Methodology: Groups of
50 snails (3-5mm) of B. t. guaibensis, F1 population of the snails from Guaíba (RS), were
individually challenged in the first experiment with 50 miracidia/snail and in the second
experiment with 100 miracidia/snail of the LE S. mansoni strain. For both infection controls
it was included groups of 50 B. glabrata specimens (6-8mm) from Barreiro de Cima, Belo
Horizonte (MG) infected with 10 miracidia/snail of the same strain, which had been kept
in the Moluscário Lobato Paraense at Centro de Pesquisas René Rachou Fiocruz (MG). As
negative controls 10 snails of each species and subspecies aforementioned were used.
The first examination of the snails, by light exposure, happened 30 days after exposure to
miracidia and then the exams were done weekly until completing 80 days of infection. The
negatives snails in the last exam were squeezed between two plates to check the presence
of S. mansoni. The LS-PCR technique was used for confirmation of presence/ absence of
the S. mansoni DNA in B. t. guaibensis snails that died along the experiment. Results:
Biomphalaria t. guaibensis showed no evidence of S. mansoni cercariae or sporocysts in
any infections while the infection rate of experiments of B. glabrata in these two infections
was 100%. The profile of the LS-PCR confirmed the absence of DNA of the S. mansoni in the
B. t. guaibensis. The mortality rate for subspecies B. t. guaibensis in the first infection was
16% and in the second was 24% and to B. glabrata the rate was 68% in the first and 34%
in the second infection. The mortality rate for the negative control of the B. t. guaibensis
were 20% in the first infection and 10% in the second and to B. glabrata was 0,0% for both
infections. Conclusions: Therefore the snails of subspecie B. t. guaibensis did not get infect
when 50 or 100 miracidia/snail of the LE S. mansoni strain were used. Future infections will
be conducted using the SJ strain, because it is more adapted to B. tenagophila.
217
gIS CHaRaCtERIzatIOn Of tHE ECOLOgICaL nICHES Of SCHIStOSOMIaSIS vECtORS
fROM tHE SUB-BaSIn dOIS RIOS RIvER, RIO dE JanEIRO StatE, BRazIL
Pablo Menezes Coelho; Paula thaise Bermudez dos Reis; Monica ammon fernandez; Silvana
Carvalho thiengo
Introduction: Watersheds are among the major influences on the distributional patterns of
freshwater species. The largest watershed in the Rio de Janeiro State is the Paraiba do Sul,
composed of several sub-basins. One of them is the Dois Rios sub-basin, which is formed by
the Negro river and Grande river, covering ten municipalities: nine in the Centro Fluminense
Mesoregion and one in the Noroestre Fluminense Mesoregion. The survey of the freshwater
gastropods of that region (2002; 2006), indicated the presence of three Schistosoma
mansoni natural intermediate hosts (Biomphalaria glabrata, Biomphalaria tenagophila
and Biomphalaria straminea) in addition to the potential vector Biomphalaria peregrina.
The occurrence of the species, when combined with thematic maps can provide input to
the knowledge of their distribution. The aim of this work is to characterize the distribution
patterns of Biomphalaria species on the Dois Rios sub-basin and compare them with those
previously reported to Noroeste Fluminense Mesoregion of the State of Rio de Janeiro.
Methodology: The molluscs were collected from different suitable snail habitats (streams,
rivers, marsh areas, drainage and sewage ditches, ponds, flood areas and irrigation canals)
and associated with thematic maps of land use, topography, clime, soil type, rainfall and
hydrography provided by GIS Results: 105 georeferenced biotopes were analyzed and B.
tenagophila was the most frequently species, found in 35 biotopes, followed by B. peregrina
(10 biotopes), B. straminea (five biotopes) and B. glabrata (three biotopes). Georeferenced
data, in combination with thematic maps provided the following Results: (1) B. glabrata
was found in seasonal forests (66,5%) and pasture (33,5%); B. tenagophila: pasture (71,5%),
secondary vegetation (14,5), urban area (11,5%); B. straminea: pasture (100%); B. peregrina:
pasture (60%) and seasonal forest (20%). (2) B. straminea was found only at low altitudes
(up to 300 meters), B. glabrata and B. peregrina occurred in altitudes from 300 to 1500
meters and B. tenagophila was found in altitudes up to 300 meters (48.5%) and from 300
to 800 meters (51.5%). (3) Two climatic types were more frequent: tropical climate with
dry winter: (100% B. straminea and 46% B. tenagophila) and temperate climate with dry
winter and warm summer (100% B. glabrata and 70% B. peregrine). (4) B. glabrata and B.
peregrina were found only in Cambisols, whereas B. straminea and B. tenagophila were
found preferentially in Ultisols. (5) B. peregrina was found in sites with1500-1750mm of
annual average precipitation; B. glabrata and B. tenagophila in sites with 1250-1500mm;
and B. straminea with 750-1000mm. (6) Streams were the most frequent biotope (100% B.
glabrata, 90% B. peregrina, 60% B. straminea and 57% B. tenagophila) and B. tenagophila
was the only species found on polluted biotope (11,4% in sewage ditch). Conclusions: B.
glabrata presented a restricted distribution, in the upper course of the Negro River, in first
order streams in Duas Barras municipality (where 8165 schistosomiasis cases were detected
from 1995 to 2009) and it was found at the same range of altitude and climate as in the
Noroeste Fluminense Mesoregion (in the Porciúncula municipality), and at the same range
of rainfall as the northeastern region of Brazil, where it is reported too. B. tenagophila was
218
found both in low and high altitudes, and it was the only species found in urban areas. It was
collected chiefly in the lower course of both the Negro river and Dois Rios river in or near
streams of first, second and third orders. B. straminea presented very restricted distribution
in the lower course of Negro river (Itaocara municipality only). In relation to topography and
climate, the results were similar to those found in the Noroeste Fluminense Mesoregion.
Regarding the rainfall, the presence of this species is similar to that of the Northeastern
region of Brazil. B. peregrina was found in three municipalities in the upper course of the
Grande River, at high altitudes and temperate clime, in areas with the highest annual rainfall
average in the State. In sum, B glabrata and B. peregrina occurred at low temperatures, high
altitudes and rainfall in less urban sites, occurring in lentic and low hydraulic conductivity
biotopes, whereas B. straminea prefers low altitude and rainfall and warm temperature. B.
tenagophila confirmed its widespread distribution in Rio de Janeiro State as well as wider
range of biotopes than the other congeneric species.
219
InfECtIOn Of BIOMPHAlARIA GlABRATA BY SCHISTOSOMA MAnSOnI In tHE
SURROUndIng SUBURBS Of SãO LUíS, MaRanHãO
andiara garcez de Souza Silva; Luciana Patrícia Lima alves; Selma Patrícia diniz Cantanhede;
Marjane Soares ferreira; nêuton Silva-Souza
Universidade Federal do Maranhão, MA - Brasil
Introduction: Humanity lives with schistosomiasis since antiquity. While in Brazil this disease having
been established during the colonial period, its first report was described only between the years
1907 and 1908 by Pirajá da Silva. In the state of Maranhao, this endemic disease has been reported
since 1920 and has a significant prevalence in several cities, causing serious disturbances to public
health. In the capital, São Luís, the peripheral neighborhoods are the most affected due to lack
of sanitation, population concentration and the presence of water collections containing large
numbers of infected snails. Among the suburbs, Coroadinho, Bom Jesus and Coroado present the
species Biomphalaria glabrata as the principal transmitter of schistosomiasis and each year the
National Health Foundation (FUNASA) records several specimens of this mollusk in the place. In
this context, the study aimed to realized a survey of the Malacological survey conducted by FUNASA
in the natural breeding neighborhoods Coroadinho, Bom Jesus and Coroado in the years 2006 to
2009. Methodology: We conducted a survey of the Malacological survey of surrounding suburbs,
Coroadinho, Bom Jesus, and Coroado, covering the period of four years (2006-2009). The annual
number of collected and positive snails for schistosomiasis was obtained from the files provided
by FUNASA of Maranhao. The Malacological survey was conducted by a team of trained health
workers of this institution, whose procedure consisted in collecting the samples with the aid of metal
tweezers and shell capture, identification of specimens by dissection and exposure of the genital
tract and finally, verification of positivity for S. mansoni by the techniques of photostimulation and
crushing. Results: In the annual survey realized in the archives of FUNASA/MA, it was observed that
in four years, a total of 4983 snails collected, 0,72% were positive for S. mansoni. The Coroadinho
district had the highest percentage of positivity. In 2006 were collected specimens in 1005 and of
these, 1,19% were positive, while that of the 565 collected in 2007, 0,70% were positive in 2008
of 885 collected, 1,01% were found positive and in 2009 of 637 collected, 0,78% were positive.
The neighborhood Bom Jesus was positive only in 2006, where the 165 snails collected obtained a
positivity of 1,81%. In subsequent years, with 165 collected in 2007, with 216 collected in 2008 and
2009 with 28 collected, the positivity was 0%. While in the neighborhood Coroado positivity was
observed only in 2006, the 135 collected, 0,74% were positive and in 2007, of 321 collected, the
positive rate was 0,62%. In the years 2008, were collected 378 and in 2009 with 483 snails collected,
the positivity was 0%. Conclusions: According to these data, we can see that the neighborhoods
are important foci in the transmission of schistosomiasis in São Luís, mainly the neighborhood
Coroadinho, since positivity was found in the four years of study. Although the rate of positivity in
neighborhoods to be considered low, these molluscs show character worrisome because they are
powerful transmitters of the parasite in the areas, since each specimen, depending on the potential
for infection, have the ability to release about 18 000 cercariae per day. The disordered occupation
of these spaces, and degrade the original ecosystem, created the ideal environmental conditions for
the emergence of numerous breeding snail in potential sites for the human infection, establishing
and maintaining the transmission of the disease.
220
MaLaCOLOgICaL SPECIES Of BIOMPHAlARIA Of CItIES In tHE JURISdICtIOn In tHE
ManagEMEnt Of REgIOnaL HEaLtH aLfEnaS-Mg, BRazIL
Rubens dos Santos vieira Júnior; Rosangela vieira Siqueira; dener Pádua Pimenta; Liliane
gonçalves vila nova; Letícia gonçalves Resende ferreira; danilo Ribeiro ferraz; Raquel Lopes
Martins Souza
Introduction: Different species of molluscs are found in the Brazilian territory which are of
great epidemiological importance in disease transmission to humans. Stand out, the snails
intermediate hosts of parasites, among which stand out those of the genus Biomphalaria,
transmitters of Schistosoma mansoni. The objective of this study was to identify species of
snails of the genus Biomphalaria, searching for the presence of parasites in these molluscs,
check the susceptibility of the snails to infection by S. mansoni and provide educational
information through lectures and distributing pamphlets to migrant workers in agricultural
enterprises import workers from endemic areas for schistosomiasis and for members of the
Association of fish farming of Alfenas. Methodology: This study was conducted through an
extension project of the University Of Alfenas UNIFAL-MG, Brazil in partnership with the
Regional Health Management Alfenas, Brazil. The study began in 2009 up to the present
date. The molluscs were collected in the cities and sent for analysis at the laboratory of
Parasitology Basic and Clinical Parasitology UNIFAL-MG, Brazil. Moreover, the interview with
workers and farmers provided guidance regarding to the collection and handling of the snails.
The snails were identified by shell morphology and internal anatomy. To analyse the presence
of parasites of the genus Biomphalaria snails were evaluated under light and shells crushing
to verify the elimination of cercariae. To cause infection, 50 snails of the genus Biomphalaria
were placed individually in a six well plate. Each well contained water with 20 miracidia of
S. mansoni. Results: There were 386 specimens of molluscs of the genus Biomphalaria in
Arceburgo city and of Fame city (MG State, Brazil). It was verified the presence of 38 specimens
of B. peregrina town of Fame and 350 B. tenagophila in the city of Arceburgo, but these
mollusks biology presented negative S.mansoni. The species identification was confirmed
by molecular methods performed at the Laboratory of Malacology of the Research Center
René Rachou / FIOCRUZ, Brazil. The 45th day after the induction of infection, the snails were
examined. However, could we not verify the elimination of cercariae. Conclusions: Once
Biomphalaria snails are found in a region rich in ponds and lakes, agricultural companies
that import labor from endemic regions for S. mansoni, the surveillance of the region activity
seems to be necessary in order prevent establishment of outbreaks for schistosomiasis.
221
MaLaCOLOgY and EnvIROnMEntaL aSSESSMEnt Of URBan OUtBREaKS In tHE
MEtROPOLItan aREa Of aRaCaJU, SERgIPE, BRazIL.
daniel Santos Oliveira; veronica de Lourdes Sierpe Jeraldo; Cláudia Moura de Melo; Álvaro
Silva Lima; Sirleide Pereira farias; Camila dantas de Carvalho; vanessa Bispo Santos;
alexandro Carvalho Silva
Universidade Tiradentes, MG - Brasil
Introduction: The quick pace of occupation of urban spaces has been reflected in the
worsening of the sanitary and health conditions of the population, including Aracaju´s
metropolitan area. Thus, the purpose of this survey was to evaluate the presence of foci
of schistosomiasis in the urban areas of Greater Aracaju/SE. Methodology: Malacological
collections were performed from 12/2008 to 06/2010 in São Cristóvão(SE) and Nossa
Senhora do Socorro(SE), the neighboring cities of Aracaju-SE. The latter collections were
made in the following neighborhoods/periods: Aruana 10/2009 to 07/2010; Coqueiral
07/2009 to 07/2010; Japãozinho 03/2010 to 07/2010 and Soledade 09/2010 à 07/2010, the
choice of sampling sites was based on information from the PCE (Programa de Combate a
Esquistossomose) which showed human cases. In order to verify the quality of the water,
this was subjected to microbiological analysis by the method of multiple tubes with values
expressed in NMP/100mL. All schistosomiasis´ foci studied were georeferenced with GPS
e-Trex Garmin ® and subsequently inserted into a database of aerial photo-map using the
ArcView Software. The malacological collections were held monthly, with sampling effort of 1
hour / 2 men, using a metal sieve (M80). The snails collected were sent to the biometric and
parasitological assessment steps (digital caliper) in the Laboratory of Infectious and Parasitic
Diseases (LDIP/ITP). The parasitological evaluation was processed according to the technique
of individual snails exposure to 60w incandescent light at a distance of 30cm from the
object. After 2 hours of observation, it was made the disposal of cercariae in a stereoscopic
microscope. After all the analysis, there were selected a few specimens of each collection
point for dissection and identification of the species found. Results: The marking with GPS of
the collecting points indicated the following coordinates: W 37 ° 4‘54 .647 “ S 11° 0‘36 .240”
(Aruana), W 37º 3‘15 .769” S 10° 52‘24 .673” (Coqueiral), W 37º 4‘21 .216” S 10 52 ‘43 .877
(Japãozinho), W 37º 8‘1 .857” S 10° 55‘9 .275” (Nossa Senhora do Socorro), W 37º 12‘21
.238” S 11° 0‘27 .460” (São Cristóvão) and W 37º 5‘5 .056” S 10° 53‘1 .738” (Soledade). In all
the collection points, the only species found was Biomphalaria glabrata. It was observed in
these sites a lack of basic sanitation, open sewers near recreation areas and homes, as well
as illegal waste dumps in various bodies of water without contact with drains and sewers.
During the analyzed period, the amount of collected B. glabrata snails was: 1022- Aruana,
2520-Coqueiral, 2044-Japãozinho, 10045-Nossa Senhora do Socorro, 5281-São Cristóvão
and 3443-Soledade. The parasitological evaluation revealed that the rate of positivity for
S. mansoni was: Aruana 8 (0.78%), Coqueiral 20 (0.79%), Japãozinho 16 (0.78%), Nossa Sra.
do Socorro 117 (1.16 %), São Cristóvão 131 (2.48%) and Soledade 14 (0.40%). From a total
of 24355 collected snails, 0.79% (306) of these were infected. The microbiological analysis
of the water showed levels of fecal pollution in the order of 3.9 x10 in Aruana, 2.8 x 104 in
Coqueiral; 1.8 x 104 in Japãozinho, 1.4 x 104 in Nossa Senhora do Socorro, 9.2 x 104 in São
Cristóvão and 1.1 x 104 in Soledade, with all these values except Aruana above CONAMA´s
222
criteria for bathing (2.5 x 103). During the studied period, Japãozinho was benefited from the
government´s sanitation projects, which has extinguished the focus of vectors. In Soledade
were built numerous buildings that occupied part of the vacant lots that accumulate water
and where the snails used to grow, which has dramatically reduced the amount of snails.
Regarding to snails´ infection, São Cristóvão has the highest rate, compared to the total
of collected/infected by the collect point, followed by Nossa Sra. do Socorro, Coqueiral,
Aruana and Japãozinho in equal proportions and ultimately Soledade . Conclusions: In the
municipality of Nossa Senhora do Socorro we observed the highest number of collected and
infected snails. São Cristóvão has the highest rate of water contamination for fecal coliform.
In terms of proportion and quantity of collected snails and the number of infected specimens,
São Cristóvão has the highest rates.
223
POPULatIOn dYnaMICS Of BIOMPHAlARIA AMAZOnICA POtEntIaL vECtOR Of
SCHISTOSOMA MAnSOnI and tHE ExOtIC SnaIL MElAnOIDES TuBERCulATuS In aPM
RESERvOIR, MatO gROSSO, BRazIL
Monica ammon fernandez; aline Carvalho de Mattos; Silvana Carvalho thiengo
Fundação Oswaldo Cruz - Laboratório de Malacologia/IOC, RJ - Brasil
Introduction: Environmental changes from water resource developmental projects affect
species richness, abundance as well as the epidemiology of water-associated diseases. Aiming
to investigate the occurrence and distribution of freshwater snails of medical and veterinary
importance in the impacted area from the Manso hydroelectric power station, Mato Grosso
State, surveys were performed from February 2002 to February 2004 and revealed the
occurrence of four gastropod families, including Biomphalaria amazonica Paraense, 1966,
potential vector of schistosomiasis. In previous papers we had pointed the susceptibility of
populations of B. amazonica from Manso dam to infection with BH strain of Schistosoma
mansoni Sambon, 1907. Epidemiological factors in addition to the pointed susceptibility
had indicated the need of quantitative analysis of freshwater molluscs. This paper presents
the results of that analysis that has been carried out since April 2009 with emphasis on
population dynamics aspects of B. amazonica and the exotic gastropod Melanoides
tuberculatus (Müller, 1774). Methodology: Collections were performed every two months
(from April 2009 to August 2010) in ten sites in APM reservoir: Manso river (three sites:
14o48’00”S, 55o40’10”W [1], 14o48’53”S, 55o35’07”W [2] and 14o51’14”S, 55o39’10”W
[3]), Palmeiras river (14o48’22”S, 55o36’32”W [4] and 14o49’06”S, 55o37’02”W [5]), Casca
river (14o56’24”S, 55o47’33”W [6], 14o57’35”S, 55o40’12”W [7], 15o00’47”S, 55o35’51”W
[8] and 15o01’38”S, 55o36’14”W [9]), and Quilombo river (15o00’50”S, 55o43’35”W [10]).
The snail population of the each site was estimated by counting the number of snail that can
be collected by one or more collectors who search the area systematically for a measured
interval of time. The method indicated the number of snails collected per man per unit of
time. Live snails were kept at the laboratory in aquaria containing deschlorinated tap water
in the Laboratório de Malacologia of Instituto Oswaldo Cruz, Fiocruz. The specimens of each
sample were preserved in Railliet-Henry´s fluid after relaxation in a 0,05% hypnol solution
for identification. Results: Specimens of five families were found (Ampullariidae, Ancylidae,
Planorbidae, Physidae and Thiaridae) and the highest species richness occurred in Manso
river (site 2): B. amazonica, Drepanotrema anatinum (d’Orbigny, 1835), Drepanotrema
depressissimum (Moricand, 1839), Melanoides tuberculatus (Müller, 1774) and Ancylidae. On
the contrary, in the Quilombo river they were obtained specimens of Ancylidae only (August
2010). Physa marmorata Guilding, 1828 and Pomacea insularum (d’Orbigny, 1835) were also
found in the sites [9] and [1], respectively. M. tuberculatus was the most frequent species in
six sites ([2] and [6]: 88.9%; [1]: 77.7%; [3] and [4]: 66.7%; and [5]: 44.4%). During the rainy
season (from December to April), it was observed the lowest abundance of M. tuberculatus:
603 specimens/man/hour (in 2009: 60 snails in April and 145 in December; in 2010: 148 in
February and 250 in April). From June 2009 to October 2009, in a single site in the Casca river
([6]) the abundance of this species was 1054 specimens/man/hour. Samples of B. amazonica
were observed in October 2009 and August 2010, in the six surveyed sites (all rivers, except
Quilombo), but never exceeding six specimens/man/hour. The occurrence B. amazonica in
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the three rivers was accidental (<25%), whereas M. tuberculatus was constant (≥ 50%) in the
Manso and Palmeiras rivers and accidental in the Casca river. Conclusions: Previous records
of B. amazonica in the reservoir were restrict to the presence of aquatic macrophytes (Pistia
stratiotes and Salvinia auriculata) that used to cover large areas. Probably, the reduced
number of B. amazonica found in this study is related to the scarce quantity of those
macrophytes present in the Casca and Manso rivers. As for M. tuberculatus, introduced
after 2004, it was the most frequent snail, and it is in the explosive phase of invasion in the
reservoir nowadays.
225
POSItIvE fOR SCHISTOSOMA MAnSOnI On BIOMPHAlARIA GlABRATA COLLECtEd In
nEIgHBORHOOd COROadInHO, SãO LUíS – Ma
Luciana Patrícia Lima alves; andiara garcez de Souza Silva; Selma Patrícia diniz Cantanhede;
Marjane Soares ferreira; Marta Martins almeida; Hallyne davinck Mesquita Moreira; nêuton
Silva-Souza; alessandra Leda valverde; Jackson Ronie Sá da Silva
Universidade Estadual do Maranhão, MA - Brasil
Introduction: Schistosomiasis is a serious public health problem in Brazil, it is present in
19 states. In Maranhão, a state of the northeast of Brazil, this endemic disease has been
reported since 1920, occurring in two distinct areas: the endemic area and the focus area.
The focus area is located in the capital’s state, São Luís, which has some neighborhoods
that present environmental and socioeconomic factors conducive to the maintenance
and transmission of the disease. Among these places, the neighborhood has peripheral
Coroadinho species of Biomphalaria glabrata as the principal transmitter of schistosomiasis
and, according to the Municipal Health Secretary, every years are reported several cases of
autochthonous Schistosomiasis mansoni in this location. In this context, the study aimed to
conduct a survey on the natural breeding malacological on Coroadinho’s neighborhood, to
see the potential for infection of B. glabrata by Schistosoma mansoni and the activity level of
this environment in disease transmission. Methodology: The natural habitat for B. glabrata,
located in the neighborhood Coroadinho peridomestic area was georeferenced using a
handheld GPS, with the geographic coordinates: S 02 º 33 ‘40.1‘‘and HO 44 º 16‘ 00.6‘‘. Soon
after the georeferencing, specimens of B. glabrata were collected, using metal tongs, over a
period of 30 minutes. The snails were transported to the laboratory of Human Parasitology,
State University of Maranhao, where they were kept in glass aquaria with dechlorinated
water and substrate, fed with fresh lettuce. Both the dechlorinated water as the substrate
were renewed once a week, placed in a proportion that depended on the number of snails.
Specimens collected, 10% were separated for the confirmation of species, which was
determined through morphological and conchological parameters based on literature. For
the analysis of positivity, the snails were once a week, over a period of forty days, isolated
in glass containers with about 5 mL of dechlorinated water, and exposed to light and heat
three 60W bulbs for one hour. Were then examined under magnification, with a positivity
indicated by the presence of cercariae of S. mansoni. Results: Collect, we obtained a total of
706 snails. Through analysis morphological and conchological, it was found that the samples
belonged to B. glabrata. The positivity of snails corresponded to about 2% This percentage
is higher than that found in a study in Alagoas, in which 0.1% was observed snail B. glabrata
positive for S. mansoni. Although the positivity rate found in this study is considered
negligible, considering the potential of natural infection (70%) of B. glabrata given in the
literature, it is possible that these animals are powerful transmitters of the parasite in the
area, as each specimen has the ability to release about 18 000 cercariae per day. It is believed
that sporadic rains during the collect have contributed to the infection of snails by miracidia,
which occurred probably through the leak of a septic tank that is partially covered the side of
the flock. Because the district Coroadinho not provide adequate health infrastructure, which
leads people to store the organic waste in pits, it is possible that in the rainy season these
pits to overflow, and the viable eggs until the rains brought by natural breeding. Besides the
226
lack of sanitation and disease carrier presence in the neighborhood, biotic (vegetation) and
abiotic (light and organic matter) have contributed to the transmission of schistosomiasis,
as they provide survival and population increase of B. glabrata, keeping thus the cycle of
the parasite. Conclusions: With these results, we find that the district Coroadinho consists
of an active focus of schistosomiasis transmission, as has environmental and biological
characteristics that contribute to the establishment and success in the cycle of S. mansoni.
Furthermore, we believe it is necessary to conduct a longitudinal study in the area, aiming to
trace the epidemiological profile of the disease and raise awareness among public agencies
to implement measures to eradicate the disease.
227
PROCESS ManEgEMEnt aIMIng tHE IMPROvEMEnt Of B. GlABRATA COLOnY
MaIntanCE at MOLUSCÁRIO dO LaBORatóRIO dE ESQUIStOSSOMOSE
ExPERIMEntaL IOC/fIOCRUz tO EnSURE tHE EffECtIvEnESS Of SCIEntIfIC
RESEaRCH In dEvELOPIng
Ronaldo de Carvalho augusto; Patrícia Machado Pinto; Clélia Christina Correa Mello Silva
Introduction: In 2008, we started a project aiming the improvement of Biomphalaria
glabrata colony maintenance in the laboratory in order to increase the yield of cercariae
in Schistosoma mansoni´s cycle specifically maintained to support experimental vaccination
studies with rSm14, the only vaccine antigen candidate against schistosomiasis selected
by WHO, and first identified by Laboratório de Esquistossomose Experimental in Instituto
Oswaldo Cruz (LEE/IOC). The LEE/IOC already has a Quality system , Environmental and
Biosafety Management which was designed and established aiming to improve the efficacy
and capability in carrying out ongoing research. The LEE/IOC has as a central project the
development of a bi-valent anti-helminthic vaccine for Schistosomiasis and Fasciolosis based
on the rSm14 antigen and it involves the creation, growth and manipulation of genetically
modified organisms. Methodology: The methodology used in this project included the
following steps: definition of design, analysis and mapping the current process, ie, (“As its
done”) future process modeling (“As it will be done”), and the implementation of the new
process being developed in the laboratory, during snail´s life cycle development stages which
are: eggs in the egg masses, recently hatched snails, young snails (onset of sexual maturity from 4 to 15 weeks old) and adult snails (15 weeks of life onwards). In addition, collaborators
training and improvements in the laboratory system. The analysis of the current process
followed by its redesign was initially performed which allowed the visualization of the
chain of added value: how the work was being done, the flow of development processes in
different stages of the cycle, and the measurement points to ensure growth of the colony
of B. glabrata for prediction of experimental tests. One of the issues being considered
would be to avoid the low productivity of the colony and consequently the decrease of
positive snails needed for completion of experimental protocols Results: After the step
“As its done” several points for improvement were identified and used to model the new
process. The improvements of the process have been implemented in four main steps,
namely: (i) traceability of the creation of molluscs and maintenance cycle of the parasite,
(ii) standardization of the breeding, (iii) organization of the dynamics of the activities and
Preparation of Reports measuring the output of moluscário and (iv) training of people.
Significant gains were achieved with the project over three years.The growth of 737% in the
population of adult snails and 242% of the population of young snails, allowing an increase
in the number of infections. There was a 37% reduction in mortality and a 59% increase in
the positivity rate of infected snails. They kept 75% more positive snails per month, allowing
an average production of 490 individual cercariae in 5mL of water. For infection of mice 35
snails positive for the elimination of cercariae are used resulting that 171.500 cercariae are
produced in 50 mL on average which permit that up to 1143 mice can be infected with 150
cercariae each. Conclusions: In conclusion, it was found that the appropriate management
of our laboratory (Moluscário) and continuous improvement of processes have a positive
228
impact on the traceability and reliability of experiments with the vaccine antigen rSm14,
such as the establishment of the colony of B. glabrata, improving the maintenance cycle
of S. mansoni under experimental conditions with proper ecological management and the
efficient use of available resources in the laboratory.
229
SUSCEPtIBILItY Of BIOMPHAlARIA TEnAGOPHIlA fROM ILHa gRandE, angRa dOS
REIS, RJ, tO InfECtIOn Of SCHISTOSOMA MAnSOnI
Monica ammon fernandez; Marta Chagas Pinto; Silvana Carvalho thiengo
Fundação Oswaldo Cruz - Laboratório de Malacologia/IOC, RJ - Brasil
Introduction: Introduction of species is a problematic issue specially in islands both
from the biological and public health standpoints. Ilha Grande, an island of the Angra
dos Reis municipality is an important site of tourism in the State of Rio de Janeiro
nowadays since its natural beauties attract thousands of tourists. Concerning to
planorbid snails the previous records to that island are: Antillorbis nordestensis
(Lucena, 1954) collected at a swampy area behind the late Penal Colony Cândido
Mendes (The Ilha Grande Prison) in 1999 and Biomphalaria tenagophila (d’Orbigny,
1835) found in a stream in Vila do Abraão (23o08’49.5”S, 44o10’13.4”W) in 2005.
Previous studies had pointed out different degrees of susceptibility of B. tenagophila
populations (from 0% to 100%) when submitted to experimental infection with SJ
strain of Schistosoma mansoni Sambon, 1907. This paper deals with the response of B.
tenagophila specimens from Ilha Grande to the SJ strain of S. mansoni, under laboratory
conditions. Methodology: The descendant molluscs of specimens from Vila do Abraão,
were individually exposed to five S. mansoni miracidia of the SJ strain. This strain was
isolated in March 1982 from naturally infected snails collected in the municipality of
São José dos Campos, São Paulo state, and it showed a good host-parasite adjustment
with the B. tenagophila populations. On the 25th day after exposure to miracidia, and
then every 5th day, the snails (12.5-2.5 mm in shell diameter) were exposed to the light
of electric lamps to characterize of the precercarial period and the infection index. The
dead specimens, as well as those that survived for 60 days without shedding cercariae,
were examined by crushing their shells. The period of emission of cercariae was
followed until death in the 30 positive specimens. Each infected snail, kept separately
in a small aquarium, was exposed to the light at least four times monthly. The aquaria
were kept at a room temperature of 24-26oC throughout the experiment. As control, 60
B. tenagophila specimens (12.5-9.5 mm) from São José dos Campos/SP were infected
with SJ strain, under the same conditions. Results: Of the 259 exposed specimens,
48 became infected (18.5%), including two died with sporocysts in the body tissues.
The results were as follows: 46 snails shed cercariae (17.7%), 191 remained negative
after examined on the 60th day (73.7%) and 22 died (8.5%) during the experiment
(two with immature sporocysts, two were negative and 18 autolysed specimens which
could not be examined). The duration of the precercarial period was 38.6±3.26 days
(mean and standard deviation), varying from 35 to 46 days. Of the 30 positive snails
kept under observation to characterize the cercarial period, one stopped cercarial
emission, showing parasitological cure at dissection. The duration of the cercarial
period was extremely variable, between five and 333 days (mean and standard
deviation: 75.1±76.8 days). The percentage survival of infected snails for more than 75
days was 30%. As to the control, 13.3% of specimens became infected. Conclusions:
Previous papers discussed the susceptibility of Biomphalaria from different South
American localities to schistosomiasis transmission, such as quantitative survey of B.
230
tenagophila and the search for the associated larval helminthes, sanitation and water
supply programme as well as health education infection with miracidia of SJ strain and
pointed out an adaptation of the parasite in each area to the respective local species
of Biomphalaria. In accordance to the results of the present paper and to Ministério
da Saúde, preventive measures should be implemented in the studied area in order
to avoid.
231
tHE MOLLUSCICIdaL aCtIOn Of tHE LatEx Of EuPHORBIA SPlEnDEnS vaR. HISlOPII
and tHE SCHISTOSOMA MAnSOnI InfECtIOn On tHE COnCEntRatIOnS Of tOtaL
PROtEInS and nItROgEn PROdUCtS In BIOMPHAlARIA GlABRATA InfECtEd WItH
SCHISTOSOMA MAnSOnI dURIng LatEx SOLUtIOn
Mariana gomes Lima; Ronaldo de Carvalho augusto; Maurício Carvalho de vasconcellos;
Clélia Christina C.Mello Silva; Jairo Pinheiro
Fundação Oswaldo Cruz, RJ - Brasil
Introduction: The levels of carbohydrates and proteins in S. mansoni infected snails,
significantly decreases during the intramolluscan larval development. In the daughter
sporocyst stage in the snail’s digestive gland, parasites require arginine for their development.
Under normal conditions, B. glabrata excretes ammonium ions, but under physiological stress
such as trematode infection, the pattern of excretion changes from ureotelic to uricotelic,
due to the acceleration in the urea cycle. The objective of this study was to determine
the alterations in total protein and nitrogen products that occur in S. mansoni infected B.
glabrata exposed to Euphorbia splendens var. hislopii latex throughout its degradation.
Methodology: We used in the study B. glabrata specimens (Belo Horizonte, BH strain) which
were individually exposed to 6-8 S. mansoni miracidium (BH strain). After establishing the
latex sub-lethal dose (LC50), B. glabrata specimens were organized in 4 groups: 1) infected
and exposed to latex (If exp); 2) infected and not exposed to latex (positive control) (PC); 3)
uninfected and exposed to latex (Not If exp), and 4) uninfected and not exposed to the latex
(NC or negative control). Two groups of 30 snails each were placed into a 500 mL glass becker
and covered properly with a perforated polystyrene lid. The infected snails were divided into
eight groups according to time or days of infection: 12 hours, 24 hours, 36 hours, 7 days, 14
days, 21 days, 28 days and 35 days. The latex solution was prepared in the same time of the
exposure and the product degradation was monitored. The snail’s hemolymph of each group
was collected by cardiac puncture using 1.0 mL syringes (B-D Plastipak@) and kept at -10°C.
The hemolymph was treated to quantify total proteins, urea and uric acid contents with
the Katal Reagentes® kits. The results were expressed in mg/dl. Results: In this experiment,
the latex lethal and sublethal concentrations were 1,0 mg/L and 0.7 mg/L. The total protein
contents in infected and uninfected B. glabrata’s hemolymph exposed to latex solution
were at low levels during all periods analyzed. The uric acid contents in uninfected exposed
B. glabrata´s hemolymph were 0.3 and 0.5 mg/dl at 12 and 24 hours of exposure, which
represent a 30% and 38% reduction in relation to NC respectively. The values observed at 7
and 35 days did not suffer high variations, but they were considered statistically significant
when the Not If exp and NC groups were compared. We observed a different uric acid profile
in the hemolymph of the group If exp. When comparing If exp with PC between 12 and 36
hours, we observed a 24% reduction in the uric acid content. Nevertheless, in the period
between 7 and 35 days an increase on the uric acid content occurred. The urea concentration
in the hemolymph of uninfected exposed B. glabrata was 22.3 mg/dl, which represent a
371% increase when compared with NC at 12 hours. On the other hand, at 36 hours this
value decreased, ranging a 75% reduction when compared with NC. The urea concentration
between days 7 and 35 was low, but at the 14th day this value was 26.0 mg/dl representing a
171% increase in relation to NC. In the hemolymph of infected and exposed B. glabrata, the
232
high levels of urea were 17.0 mg/dl (12 hours), 13.0 mg/dl (7 days) and 19.0 mg/dl (28 days).
Conclusions: Under physiological stress conditions, the urea pathway enzyme’s catalytic
activity is accelerated. The variation in the nitrogen products showed that the urea increase
occurred when the uric acid level declined. This suggests a change in the excretion pattern,
moving from ureotelic to uricotelic. The present study showed the possibility of using a
natural product, E. splendens var. hislopii latex, in a sublethal dose for selective control of
infected snails showing a promising method to control schistosomiasis transmission.
233
an OUtBREaK Of aCUtE SCHIStOSOMIaSIS In tHE SOUtHEaSt Of BRazIL: I.
EPIdEMIOLOgICaL aSPECtS
Sandra Costa drummond; Izabela voieta; alba Otoni; Silvana Romano; José f.vilela; Jeann
vilela; fernando J. Silva; Wenderson Bassoli; Patrícia Passos Botelho; Carla Maria Ligeiro;
Carlos Maurício figueiredo antunes; José Roberto Lambertucci
Secretaria de Estado de Saúde de Minas Gerais, MG - Brasil
Introduction: Schistosomiasis is an endemic disease in Minas Gerais state being currently
transmitted in 523 municipalities. São João del Rei (SJDR), a city in the southeast of Minas
Gerais State, has 85,503 inhabitants; except for Matozinhos, a small district of SJDR, the disease
transmission has not been reported there since 1971.The aim of the present investigation is
to report an outbreak of acute schistosomiasis occurring in a new area of this city, named
Theodore Colony. Methodology: From December 2009 to March 2010, 40 individuals who
have always lived in SJDR bathed in a natural swimming pool supplied by a local brook in a
country estate in the periphery of SJDR. Two weeks later a patient with myeloradiculopathy
was attended at a reference center for schistosomiasis in a General Hospital of Belo
Horizonte, Brazil. A specialized team from the Minas Gerais State Health Department visited
the site after being informed about the outbreak; all of those exposed were submitted to
clinical, laboratory and abdominal ultrasound examinations. For malacological survey and
identification of possible foci of contamination, 200 snails were collected: the investigation
was made at each 50 meters along the stream waters, at this new transmission area. The
collected snails were sent to the a reference laboratory of malacology for morphological
identification. Coproscopical analysis of a stool sample (analyzed by the Kato-Katz technique
and a sedimentation method) for each participant was performed. Results: The patients age
ranged from 2 to 82 years (mean: 33.5; median: 29) and 21 were male (51%). Sixteen out of
41 individuals (39%) developed symptoms of acute schistosomiasis and stool examination
revealed eggs of S. mansoni in 15 (36.5%). The main symptoms were: cercarial dermatitis
in 13 (31.7%), cough in 10 (24.4%), diarrhea in 9 (22.0%), fever in 7 (17.0%), bloody stools
in 7 (17.0%), lower limb weakness in 1 (2.4%). Five patients were hospitalized and one had
myeloradiculopathy. The latter was the first patient diagnosed with acute schistosomiasis
and she indicated that other patients in her hometown were also infected on the same
occasion. Three presented pulmonary involvement and one had severe gastrointestinal
symptoms. One patient refused treatment and the others received praziquantel (50 mg/
kg, body weigth). Biomphalaria glabrata was the only species of snail identified in the area:
all of them were dead and therefore it was not possible to isolate cercariae of the study
specimens. The Theodore Colony is not a perennial area of schistosomiasis transmission.
Our hypothesis is that construction workers, who recently moved in to build up new houses
above the site of water contact, came already infected from schistosomiasis endemic areas
(to be confirmed), and contaminated the watercourse. Conclusions: The first patient who had
acute schistosomiasis with myeloradiculopathy triggered the investigation of this outbreak in
Theodore Colony, SJDR which is not an endemic area. Our study suggests that non-endemic
areas may occasionally become the source of periodical transmission of schistosomiasis with
the surge of new epidemics of acute schistosomiasis.
234
aSPECtS RELatEd tO tHE dEatH BY SCHIStOSOMIaSIS, OBtaInEd fROM
InvEStIgatIOnS WItH faMILY In JaBOataO dOS gUaRaRaPES-PE, JUnE 2010
francine nesello; tania gomes de Carvalho; José Holanda dos Santos neto; Rodriga Maria
zovkar de Souza; José alexandre Menezes da Silva
Introduction: Jaboatao Guararapes is a municipality located in the metropolitan area of Recife
- Pernambuco - Brazil, and has approximately 700 000 inhabitants. Divided into 27 districts
and 06 regional authorities, the municipality is endemic for schistosomiasis and presents high
prevalence and significant mortality. The objective of this study is to know environmental factors
and individual habits that has influenced the mortality from schistosomiasis. Methodology: This
study was conducted at Jaboatao dos Guararapes. This is a descriptive study with data obtained
from the municipal base of the System on Mortality Information (SIM), relative to January 2006
- June 2010. To obtain the environmental variables and individual habits, we applied a semistructured questionnaire with questions about the environment where the patient lived, sought
to know the probable site of infection and information regarding the assistance provided to the
individual during the period of the disease. These questionnaires were applied with close family
members to the selected case. It was selected for convenience, in the SIM, ten deaths to be
investigated. In the analysis software was used TABWIN, Microsoft Office Excel 2007 and EpiInfo
windows 3.5.1. Results: Considering the Schistosomiasis as the underlying cause (ICD-10: B65.1),
there are 53 cases of deaths in Jaboatão dos Guararapes, from January 2006 to May 2010. Of
these, 28 (53%) were male and 25 (47%) females. The median age at time of death was 65 years,
ranging 32-96 years. Approximately 35% (the 19th) of the cases occurred before age of 60 years
old. By place of residence, were homogeneously distributed in five regions, except the regional
04, which presented no death. The school could not be evaluated because in 58% (31) from death
certificates, this field was ignored by those who filled out. In only six of the ten deaths selected for
investigation, we found the family at the address provided. Among the investigated deaths 100%
(6) lived in urban areas, 83% (5) of households had piped water, 83% (5) of dwellings, sewage
was thrown into a open channel, and had reported that the patients who died had contact
with contaminated water while leave their homes, especially on rainy days when the channel
overflows and flooded streets. Families also reported that 50% (3) of patients had the habit of
fish and 67% (4), the habit of swimming in rivers and lakes in the region. Regarding medical care,
only half of the cases 50% (3) conducted a stool test before the final stage of the disease in 100%
(6) reported that the families had at least one of the characteristic symptoms such as diarrhea,
vomiting with blood and ascites fluid accumulation in the abdominal cavity. Only 33% (2) looking
for the basic unit of health, albeit belatedly, and 100% (6) were admitted to hospital before
death. Conclusions: We observed that some prominent factors influencing the occurrence of
death, such as the late discovery of the disease, difficulty in clinical diagnosis and epidemiological
study in primary health care and lack of continuous care to patients. It is also concluded that in
general, people know the disease, its transmission and the majority of presenting symptoms, but
do not identify parasitism as severe symptoms by Schistosoma mansoni. Challenger: Therefore,
we emphasize the important role of integrating primary care with the residents of sites conducive
to infection by carrying out health education, coproscopic surveys and providing treatment in full.
235
CaRBOHYdRatE MEtaBOLISM In BIOMPHAlARIA GlABRATA InfECtEd WItH
SCHISTOSOMA MAnSOnI and ExPOSEd tO LatEx fROM EuPHORBIA SPlEnDEnS vaR.
HISlOPII dURIng ItS dEgRadatIOn
Mariana gomes Lima; Ronaldo de Carvalho augusto; Clélia Christina Correa Mello Silva; Jairo
Pinheiro
Introduction: Schistosomiasis is an important human parasitic disease and has the snail
Biomphalaria glabrata as its main intermediate host in Brazil. Many molluscicides have been
used in the control of schistosomiasis around the world but one of the most promising is the
crude latex from Euphorbia splendens var. hislopii. The latex obtained from this plant showed
under laboratory and field conditions that it attends to queries needed to be used as natural
molluscicide. Energy main resources used by pulmonates snails are glycogen and galactogen
present in the digestive gland and mass cephalopodal and in the albumen gland of mollusks,
respectively. The main of objective of this study was the evaluation of the physiological
impact of latex from E. splendens var. hislopii over the carbohydrate content of B. glabrata
infected by S. mansoni through the degradation time of the latex Methodology: In order to
do this study, specimens of B. glabrata (Belo Horizonte, BH strain) were individually exposed
to 6-8 miracidium of S. mansoni (BH strain) isolated from eggs eliminated in the faeces
of Swiss albino mice, in our laboratory at the Oswaldo Cruz Foundation in Rio de Janeiro.
After the sub-lethal dose (LC50) was established, specimens of B. glabrata were organized
in 4 groups: 1) infected and exposed to latex (If exp); 2) not infected and not exposed to
latex (positive control) (PC); 3) not infected and exposed to latex (Not If exp), and 4) not
infected and not exposed to the latex (NC or negative control). Groups of 60 snails each were
placed into a 500 mL glass becker and covered properly with a perforated polystyrene lid.
The infected snails were divided into eight groups according to time or days of infection: 12
hours, 24 hours, 36 hours, 7 days, 14 days, 21 days, 28 days and 35 days. Each 60 specimens
group was exposed to a recently prepared latex solution for 24 hours, and the action of
the product degradation was monitored. The hemolymph from snails of each group was
collected by cardiac puncture using 1.0 mL syringes (B-D Plastipak@) and kept at -10°C until
the moment of glucose determination.) and results were expressed as mg/dL. The digestive
gland (DG) and cephalopedal mass (CM) were separated, weighed and frozen until glycogen
extraction and determination. The glycogen content was expressed as mg of glucose/g of
tissue, wet weight Results: The lethal and sub-lethal concentrations were 1.0 mg/L and 0.7
mg/L in this experiment. The glucose content in the hemolymph of infected and exposed
snails was 35.5 mg/dL at the 7th day, which represents 433% of the glucose content of the
PC. In the uninfected and exposed group the glucose content was 8.8 mg/dl. This result
shows a significant reduction and represents 30% of the NC. The significative alterations on
the carbohydrate contents in the exposed snails were verified from the first day to the 14th
day after that the aqueous-latex solution was prepared. The glycogen contents in the CM
and DG increased 68% and 46% when compared with PC group at the 12 hours. At the 7th
and 14th days, the glycogen contents in the CM decreased 16% and 20% in relation to the
PC. In DG, the reduction of glycogen content was significant, with maximal reduction during
the different periods of time analysed here, with 0.3 and 0.5 mg of glucose/g of tissue, wet
236
weight, which represent 17 % and 22% reduction in relation to the PC. After the 21st day
and, at the end of the observation period, the glycogen contents in CM and in DG showed
values near to the glycogen contents of PC. In uninfected and exposed mollusks the glycogen
contents in the CM at the 7th and 14th days, decreased 17% and 14%, respectively, when
compared to NC. In DG, the reduction of the glycogen contents was significant, at the same
period, and around 25% and 14%, respectively. The same behavior was observed at the 21st
and at the end of the observation (for both treatments groups) when the glycogen contents
in CM and in DG was similar to those observed for PC. Conclusions: The infected and exposed
mollusks showed a decrease in the glycogen contents much higher than the uninfected and
exposed mollusk at this period, displaying a slower recuperation. However, the association of
factors such as stress, infection and poisoning by the latex gave rise to significantly different
results, when comparing to mollusks only exposed to the latex solution. Consequently, we
conclude that the action of the latex can be more efficient over individuals infected with
larvae of Schistosoma mansoni than otherwise.
237
CO-InfECtIOn Of SCHISTOSOMA MAnSOnI WItH HIv/aIdS In EndEMIC POPULatIOn
In tHE StatE Of aLagOaS
danielle Correia gama; Rozangela Maria de almeida fernandes Wyszomirska; Maria Sônia
Correia alves; Luciano fernandes Pereira; Roberta Maria Pereira albuquerque de Melo;
dńarte Hermogenes Bastos; Hugo Cabral tenório; andrei Leite gazzaneo
Universidade Estadual de Ciências da Saúde de Alagoas, AL - Brasil
Introduction: Schistosomiasis mansoni is an endemic disease, may progress to severe
clinical forms, installation of portal hypertension. In Brazil, it is more prevalent in the states
of Maranhão, Ceará, Rio Grande do Norte, Paraíba, Pernambuco and Alagoas. It has been
suggested that HIV and parasitic infections may interact affecting each other, altering the
natural history of disease, either by preventing the rapid diagnosis or reduce the effectiveness
of antiparasitic treatment possible.Identify the frequency of co-infection with Schistosoma
mansoni with HIV-1 and HIV-2 in an endemic region of the state of Alagoas. Methodology:
An inquiry was conducted in three locations coprological (Island Angelita, Lourenço de
Albuquerque and farm Riachão) in the city of Rio Largo, State of Alagoas. Research coprological
was performed using the Kato-Katz (02 slides per person), from November 2007 to August
2008. The following individuals with S. mansoni were submitted to blood collection without
the use of anticoagulants for the determination of qualitative immunochromatographic tests
that detect antibodies to HIV-1 and HIV-2. Results: We analyzed 3030 samples of faeces. Of
these, it was found that 242 (8.0%) were positive for S. mansoni (mean age 23.77 ± 13.43
years (minimum 04 and maximum 68 years) for males and females with 27.61 ± 17.34 years
(minimum 02 and maximum 79 years) . The test for HIV-1/HIV-2 was no reagents in all
samples. Conclusions: The areas along the rivers were analyzed, considered risk areas, but
were not found in patients co-infected with HIV-1/HIV-2. The results were not decisive for an
assessment of the impact and should be a study population.
238
EffECt Of PHYSAlIS AnGulATA (CaMaPÚ) ExtRaCt and fRaCtIOn In ExPERIMEntaL
SCHIStOSOMIaSIS ManSOnI.
José augusto albuquerque dos Santos; Ivone Maria Ribeiro; therezinha Coelho Barbosa
tomassini; neusa araújo; ana Karine Sarvel de Castro; Luis Cláudio Muniz-Pereira; antonio
Henrique almeida de Moraes neto; naftale Katz
LAPSA, Instituto Oswaldo Cruz, Fiocruz, RJ - Brasil
Introduction: The genus Physalis, belongs to the Solanaceae family and includes about 120
species with herbal characteristics and perennial habits. In Brazil, Physalis angulata L. grows
in abundance in the North and Northeast, where it is popularly known as Camapú and used in
folk medicine. In previous studies, we observed their molluscicidal activity against Biomphalaria
tenagophila. Our goal was to determine the plant´s ethanol extract effect and pool of stem
Physalins influence on the development of schistosomiasis in mice. Methodology: The plant
extract and pool of stem Physalins were supplied by Farmanguinhos, FIOCRUZ. Before, during
and after infection with S. mansoni cercariae, BH strain, each mouse was phytotherapeutically
treated at the Laboratory of Schistosomiasis/ CPqRR. Infection was conducted through a 50
min exposure of each mouse individually to a solution containing 100 cercariae. In the three
experimental groups, each composed of 14 animals, the first and second were treated with
100 mg/kg and 300 mg/Kg of ethanolic extract aqueous solution, respectively and the third
with 300 mg/kg of the pool of stem Physalins, all three solutions elaborated from plant extract
and pool dissolved in DMSO and afterwards in PBS. The control group of also 14 animals was
exposed to only a pure DMSO/PBS solution. After 45 days, the time required to complete the
S. mansoni life cycle, perfusion for worm recovery from the liver and mesentery ensued. All
animals were humanely sacrificed by cervical dislocation according to the animal code ethics
committee protocol of FIOCRUZ. The liver and mesentery collected at necropsies were weighed
and measured by oogram. Results: In the control group, the total average number of S. mansoni
retrieved per animal was 10.14 in the mesentery and 1.43 in the liver. Considering the original
infection of 100 cercariae per animal, in the mesentery worm recovery was 31% male, 28.2%
female and 20.4% couples and in the liver, 60.1% male, 30% female and 4.9% couples. The first
experimental group treated with 100 mg/kg of plant ethanol extract yielded a total average
per animal of 12 S. mansoni recovered from the mesentery and 2.57 from the liver. In the
mesentery, the recovery percentage was the same for both males and females (28.6%) and
21.4% for couples, while in the liver, it was 38.9% for both males and females and 10.9% for
couples. For the second experimental group, treated with 300 mg/kg, the average recovery of
worms was 13.82 per animal in the mesentery and 3 in the liver. In the mesentery, recovery was
32.2% for both males and females and in the liver, 36.3% male, 45.3% female and 9% couples.
The third experimental group, treated with pool stem Physalins, had an average total of 17 S.
mansoni retrieved per animal in the mesentery and 3.1 in the liver. In the mesentery, recovery
consisted of 31.9% male, 30.9% female and 18.6% couples and in the liver, 62.3% male and 38%
female. Conclusions: The plant extract and pool of stem Physalins in the adopted doses did not
significantly reduce disease development, although the female worm recovery percentage in
the liver for the third experimental group, tested with the pool of Physalins, decreased to half,
when compared to males. These results encourage further studies on the phytotherapeutic
effect on S. mansoni larvae.
239
EffECt Of tREatMEnt WItH PRazIQUantEL In COgnItIvE PERfORManCE Of
CHILdREn InfECtEd BY S. MAnSOnI In a RURaL aREa Of vaLE dO JEQUItInHOnHa,
MInaS gERaIS
Márcia Christina Caetano de Souza; Mery natali Silva abreu; Humberto f.O. Quites; Leonardo
f. Matoso; Jorge gustavo velasquez-Melendez; Helmut Kloos; Rodrigo Correa-Oliveira;
andrea gazzinelli
Universidade Federal de São João Del Rei, MG - Brasil
Introduction: Schistosomiasis can impact the health of children, causing malnutrition,
anemia and cognitive deficit. It is know that antiparasitic drugs may help to improve
health conditions of individuals and also their cognitive performance. This study aimed at
evaluating the effect of praziquantel treatment on the cognitive performance of children
previously infected with S. mansoni who live in Caju and São Pedro do Jequitinhonha, rural
area in Jequitinhonha Municipality, Minas Gerais. Methodology: Two hundred and one
(201) children 6-10 years old participated in this study, all living in Caju and Sao Pedro do
Jequitinhonha rural district in Jequitinhonha Municipality. Data were gathered in 2 phases.
First, a parasitological examination was performed on all children using the Kato-Katz
method and their height and weight were measured to evaluate nutritional status. Cognitive
assessment was carried out using the Raven Test and two Digit subtests (WISCIII Code
and Aritmethic test). All children’s parents answered a demographic and socioeconomic
questionnaire. After that, infected children were treated with praziquantel and albendazol.
One year (12 months) after treatment, the nutritional measurements and cognitive tests
were repeated. Results: The prevalence of schistosomiasis was 70.6%, of hookworm 18.5%
and of ascaris 2.6%. The percentage of children with acute malnutrition before treatment
was 3.5% and with chronic malnutrition 11.4 %, with overweight and obesity 7.0%. Cognitive
performance showed that most children were under the average rate in both tests but there
was an increase in the cognitive scores after treatment, apart from being infected or not
during the first phase. Multivariate analysis showed that children who achieved the higher
scores all lived in Caju (OR=2,10 CI 95%=1,07-4,11) in houses with family heads having 4
years or less of schooling (OR=2,93 CI 95%=1,26-6,81). Children living in Caju also showed
a better performance on the subtest Code (OR=2,21 CI 95%=1,02-4,81). Children 9-10 years
of age (OR=2,48 CI 95% 1,07-5,76) also showed a significant improvement on this test. The
highest difference in the body mass index was associated with the improvement in the Code
Test (OR=1,12 CI 95% 1,01-1,23). Higher educational level of mothers was directly related to
better performance of their children on the Raven test (OR=4,28 CI 95% 1,32-13,87). Children
with better socioeconomic conditions showed improved cognitive performance on the Raven
Test (OR=5,17 CI 95% 2,12-12,59). Conclusions: The results of this study show that treatment
did not have an effect on the cognitive performance of children previously infected with
S. mansoni. However, environmental and biological factors may impact children’s cognitive
performance. Educational health programs are recommended to help families manage their
children’s health care and also to stimulate their normal developmental. Financial support:
CNPQ, FAPEMIG, NIH-ICIDR Grant 1R03AI071057-01, CAPES
240
EPIdEMIOLOgICaL aSPECtS and gEOgRaPHICaL dIStRIBUtIOn Of SCHIStOSOMIaSIS
and HELMIntHES In SERgIPE StatE aCCORdIng tO tHE data Of tHE SCHIStOSOMIaSIS
COntROL PROgRaM (PCE)
Carla virginia vieira Rollemberg; Cybele Maria Bomfim Santos; Marília B. Lemos Silva; acácia
M.B. Souza; Ângela Maria da Silva; José antônio Pacheco de almeida; Roque Pacheco de
almeida; amélia Ribeiro de Jesus
Introduction: Schistosomiasis is endemic in Brazil, with high prevalence in the state of
Sergipe, despite the development of the Schistosomiasis Control Program (PCE). According
to the Ministry of Health, Sergipe state presents a high prevalence. with an average of 17.3%
between 1980 to 1989, the second highest in Brazil, smaller only of the state of Alagoas.
In the period 1990 to 2002, the average prevalence in the state was 17.7%, well above the
national average of 9.2%. Methodology: We revised data from Sergipe’s PCE from 2005 to
2008. On average, 57% of municipalities have sent data to the PCE in these four years. We
created a database on a spreadsheet from the software ACCESS and analyzed the frequency
and geographic distribution of infections by S. mansoni and other intestinal parasites. These
data were exported to the software SPRING 5.0.5 for georeferencing and preparation of
thematic maps of spatial and temporal distribution by year of evaluation. Results: In 2005,
13.61% (14471/106287) of the exams were positive for S. mansoni, 11.16% (16196/145069)
in 2006, 11.77% (10,220/86,824) in 2007 and 10.56% (8,329/78,859) in 2008. The analysis
of maps showed high prevalences of disease in the municipalities of Ilha das Flores, Santa
Rosa de Lima, Santa Luzia do Itanhi and São Cristóvão. The proportion of people treated
after positive exams for S. mansoni was 54.5% (8471/14469) in 2005, 74.4% (12106/16267)
in 2006, 73.5% (10897/14828) in 2007 and 77.9% (6490/8334) in 2008. Socioeconomic risk
factors and environmental impacts have been associated with S. mansoni and helminthes
infections We decided to build on these analysis on the 2007 data, considering these more
reliable due to the larger number of municipalities reported. That year, the prevalence of
S. mansoni was 14.4% and of other parasites was 22.3% for A. lumbricoides, 6.1% for
hookworm, and 1.2% whipworm. Furthermore, we evaluated the association between the
frequencies of these parasitic diseases with social and development indicators of the different
municipalities, according to data from the Brazilian Institute of Geography and Statistics
(IBGE) and the Department of Water Resources (SRH). We found that cities with prevalence
of schistosomiasis higher than 15% have lower levels of sewerage (Hygiene Index), p =
0.05. Additionally, municipalities that prevalence of hookworm was higher than 10% have a
lower HDI education, p = 0.04. Conclusions: The study showed that the PCE data has a poor
coverage of municipalities in the state of Sergipe. The data also suggested an association of
the occurrence of parasitic diseases with poor sanitation conditions and educational levels
of different municipalities. Schistosomiasis is a focal disease, the spatialization techniques
used in our study should be incorporated into the current methodology used by the
Health Surveillance Secretariat to optimize the determination of areas of risk and financial
investiments to control schistosomiasis. We highlight the importance of greater control of
environmental risk factors and education, in an attempt to reduce the prevalence of these
parasitic diseases.
241
EPIdEMIOLOgICaL aSPECtS Of SCHIStOSOMIaSIS In tHE LandLESS RURaL
genilde gomes de Oliveira; angela Maria da Silva; Izabel Cristina Okuveira Lima; Jenisson
Oliveira Conceição
Introduction: Schistosomiasis is a water-borne disease caused by Schistossoma mansoni and
its clinical evolution can vary from asymptomatic forms to severe conditions. Schistosomiasis
mansoni is a global endemic disease, occurring in 74 countries and territories of three
continents. To verify through a parasitological examination of fecal samples the occurrence
of Schistossoma mansoni among landless rural workers and their families in settlements in
riverside cities from the south region of the State of Sergipe. Methodology: It is a quantitative
cross-sectional approach study conducted on landless rural workers from the south region
of the State of Sergipe. A physical examination was performed on 822 landless rural workers
and 601 of them had a parasitological examination of fecal samples to confirm the infection.
Results: A 4.3 % prevalence of Schistosomiasis among landless rural workers was found
with a positive result for the infection in 61.5 % among the 13 settlements studied. In a
100% of positive cases the clinical form of the disease found was the chronic intestinal.
The population portrayed is a low-income community, living in floor soil ground four-room
households (living-room, bedroom, bathroom and kitchen). The literacy level in these
communities is low. In these populations 73.3% of the citizens had attended elementary
school, 16.3 % of them are illiterate and 10 10.4% attended high school or higher education.
In all the settlements there is poor sanitary condition, no tap water and sewer system.
Conclusions: Occurrences of Schistosomiasis mansoni were evidenced in 8 (61.5%) of the
Landless Rural Workers settlements studied and 26 (4.3%) of them had a positive result for
the disease, although the prevalence of Schistosomiasis among landless rural workers from
the south region of the State of Sergipe is relatively low in comparison to the whole state
rates and in some cases from the south region. The predominant form of the disease is the
chronic intestinal possibly due to the short period of exposure to this environment and to
the characteristics of the migratory population. The socio environmental condition of the
population is practically the same: low literacy level, predominance of rural work, lack of
sanitation or sewer system and few of the rural workers have access to tap water. It was
evidenced a very distinct prevalence between the settlements despite the resemblance
of the socio environmental conditions, possibly due to differences concerning the deposit
water to which this population is currently exposed.
242
EPIdEMIOLOgICaL InvEStIgatIOn On an aUtOCHtHOnOUS CaSE
SCHIStOSOMIaSIS In tHE aPIPUCOS LagOOn, CItY Of RECIfE, PERnaMBUCO
Of
Constança Simões Barbosa; Elainne Christine Souza gomes; fabio Melo; Onicio Batista Leal
neto; Louisiana Quinino; Mariana Melo; Lidya angelo Bezerra; Karina Conceição araujo
Centro de Pesquisas Aggeu Magalhães Fiocruz, PE - Brasil
Introduction: In Pernambuco, new foci of vector molluscs for schistosomiasis are being
identified in the Metropolitan Region of Recife (MRR), where the lack of sewage disposal
systems creates favorable conditions for transmission of the disease. In 2008, researchers
and collaborators from the Schistosomiasis Laboratory of CPqAM/Fiocruz, undertook
a Scientific Expedition in order to update the information on foci of transmission on the
coast and in the MRR. On that occasion, snails of the species Biomphalaria straminea
infected with Schistosoma mansoni were found in the Apipucos Lagoon, which is located
in an upmarket district of the city of Recife. The current use of this lagoon has become
a source of conflict, given that it is used for discharge of solid waste and sewage coming
from the districts that surround it, while at the same time, it serves as a recreational area
for the local inhabitants, who go fishing, bathe and relax there. The aim of this study was
to survey indicators for biological and environmental risks and investigate human cases
relating to schistosomiasis transmission in the Apipucos Lagoon. For this, the following were
performed: (1) georeferenced mapping, to delimit the study area and construct thematic
maps highlighting the locations and extent of environmental and biological risk factors; (2) a
malacological survey to determine the points of fecal contamination, and to gather, identify
and examine vector snails; (3) a coprological survey among individuals who frequented the
lagoon. Methodology: 1. To map and spatially locate the points, the absolute method for
instantaneous positioning of points was used. Data were gathered by means of a Garmin
GPS (Global Positioning System) receiver. The digital mesh for the Apipucos Lagoon was
constructed using the MapInfo Professional 6.0 software. The attributes (snail gathering
locations, foci detected and points of fecal contamination) were processed and organized
in a Microsoft Excel 2003 spreadsheet. The field data recorded using the GPS receiver were
transferred through a universal serial port (USB) into a computer, where the data were
interfaced with the GPS Track Maker (GTM)® software, to adjust the templates. The Corel
Draw 13 software was used for esthetic adjustments to the map and inclusion of elements
such as the points of the compass and scale bars. 2. The malacological survey identified
the points of fecal contamination within the environment , and these were demarcated
as gathering locations, where snails were caught every month, between March 2009 and
June 2010. Determination of whether the snails were positive for Schistosoma mansoni
was performed (1) by means of conventional methodology, in which snails were exposed
to light for one hour and thirty minutes, at a mean temperature of 27ºC; and (2) by means
of a molecular approach (nested PCR) to detect the presence of DNA of the parasite S.
mansoni, when it was not possible to determine whether they were positive by means of
the conventional technique. 3. The coprological survey was carried out using an intentional
sample formed by 180 students aged 7 to 14 years from the school Colégio Marista Nossa
Senhora Conceição, who lived near to the Apipucos Lagoon. The fecal material was collected
in June and July 2010, and the material was analyzed by means of the quantitative Kato-
243
Katz method. Results: The coprological survey found that one male patient (12 years of age)
was parasitized with Schistosoma mansoni, with a low parasitic load. An investigation into
whether the case was autochthonous was conducted by means of interviews with members
of this child’s family, who gave assurances that the child never went far from that locality
and that he bathed in the lagoon every day. The malacological survey gathered 209 snails
of the species Biomphalaria straminea, which did not eliminated cercariae by means of the
conventional technique but the molecular approach (nested PCR) revealed the presence of
DNA of S. mansoni in three batches of snails that were examined, which thus indicated that
there were three focal points around the Apipucos Lagoon. Conclusions: The malacological
diagnosis plus the autochthonous human case made it possible to confirm that active
transmission of the disease was taking place in that environment.
244
EPIdEMIOLOgICaL StUdY Of SCHIStOSOMIaSIS ManSOnI InfECtIOn In tWO
COUntIES Of JEQUItInHOnHa and RIO dOCE vaLLEYS In MInaS gERaIS
Maria José Conceição; aline Eduardo Carlôto; fabiana Euzébio gonçalves Euzébio; Eric vinaud
de Melo; Iran Mendonça da Silva; natália Machado da Silva; José Borges Pereira; nelson
gonçalves Pereira; José Rodrigues Coura
Faculdade de Medicina – UFRJ / Fiocruz, RJ - Brasil
Introduction: Surveys on the natural history of infection with Schistosoma mansoni in two
counties of Minas Gerais have contributed to elucidate the parasite-host interaction and
its effect on the main prevention measures regarding to schistosomiasis. Methodology:
Two counties in Minas Gerais, Brazil were studied: Comunidade São João (Itaobim), in the
Jequitinhonha Valley (area I), and Capitão Andrade, in the Rio Doce Valley (area II). The
diagnosis of schistosomiasis was based on parasitological stool tests. The specimens of
´Biomphalaria´ were collected in the main streams of both counties, they were classified and
evaluated according to the indexes of infection. In the clinical classification, three groups were
considered: type I- intestinal form, type II- hepatointestinal form, and type III- hepatosplenic
form, the last one was confirmed by abdominal ultrasound of the patients. Infection
prevalence was determined by three parasitological examinations by Lutz´s method, and
Kato´s method modified by Katz et al. (1972), and infection burden was based on the number
of S. mansoni eggs per gram of faeces. Results: Biomphalaria glabrata was the intermediate
host in these areas, with a positivity rate of cercarial elimination of 1.2% in the area I, and
1.3% in the area II. From a total of 393 inhabitants of Comunidade São João, the prevalence
rate of schistosomiasis infection was 26,3%., of which 69.2% were classified as intestinal
form, 28.7% as hepatointestinal, and 2.1% as hepatosplenic form. In Capitão Andrade, from
a total of 349 inhabitants the prevalence rate reached 19.9 %, of which 70.2 % were type I,
26.5% type II, and 3.3% type III. Conclusions: The high prevalence of schistosomiasis infection
in the two counties of Minas Gerais with cases of hepatosplenomegaly recurrences requires
a thoroughly review of the control measures adopted at these areas aiming to avoid either
the transmission as the reinfection process.
245
EStIMatIvE Of SCHIStOSOMIaSIS POSItIvE IndEx fOR MInaS gERaIS StatE, BRazIL,
USIng MULtIPLE REgRESSIOn
Ricardo José de Paula Souza e guimarães; Corina da Costa freitas; Luciano viera dutra; Sandra
da Costa drummond; guilherme Corrêa de Oliveira; Omar dos Santos Carvalho
Introduction: Schistosomiasis caused by Schistosoma mansoni is a disease conditional on
the presence of mollusks of aquatic habits of the genus Biomphalaria. In Brazil, there are
three species of Biomphalaria (B. glabrata, B. tenagophila and, B. straminea) that have been
being found naturally infected by S. mansoni. B. glabrata is of great importance, due to its
wide geographic distribution, high infection indices and effectiveness in the schistosomiasis
transmission. In endemic areas, large concentrations of these snails, together with other
risk factors, favor the existence of localities with high prevalence. Schistosomiasis is a
disease determined, in space and time, from environmental factors such as vegetation,
land use, water collections, and others variables. Geographic Information System (GIS) is
a tool for identifying and defining, respectively, factors and possible risk areas to indicate
a better distribution of resources and a more appropriate direction for the control of
schistosomiasis. The main objective is to establish a relationship between schistosomiasis
positive index and the environmental, socioeconomic, meteorological and remote
sensing variables, in the state of Minas Gerais, Brazil, through multiple linear regressions.
Methodology: The study area includes the entire state of Minas Gerais. Data from the
Schistosomiasis Control Program (PCE) in 1,590 localities (Positivity Index - Ip), results of
malacological surveys of the Laboratory of Helminthiasis and Medical Malacology of the
Rene Rachou Research Center (CPqRR/Fiocruz-MG), as well as socioeconomical (SNIU and
IBGE), meteorological (INPE), environmental (CPRM, IBGE and João Pinheiro Foundation)
data and variables derived from remote sensing (NASA) were used in this study. Multiple
linear regressions (form of regression analysis in which data is modeled by a least squares
function which is a linear combination of the model parameters and depends on one or
more independent variables) were applied in the schistosomiasis positive index. Results:
A Database, GeoSchisto (http://www.dpi.inpe.br/geoschisto/), was created containing
all variables used in this study. Using meteorological and environmental variables that
characterize the habitat of the Biomphalaria snail, the State of Minas Gerais was divided
into four regions using the Skater algorithm of regionalization. The information of the
existence of B. glabrata and other variables were used together in multiple regression
models in Minas Gerais (global model) and regionalized areas (regional model) to estimate
Ip (localities). In all models the presence of B. glabrata, sanitation, vegetation index and
temperature were the most important variables. Results of the regression models show
that regionalization improves the estimation of the disease in Minas Gerais. Based on this
model, a schistosomiasis risk map was built for Minas Gerais. The averages interpolator
was used in the regional model for municipalities to indicate possible local transmission of
schistosomiasis. Conclusions: This study shows the importance of joint use of geographic
information systems (GIS) and remote sensing (RS) to study the risk of schistosomiasis.
Moreover, it can be concluded that the combined use of GIS and statistical techniques
allowed the estimation of schistosomiasis positive index. Results of the regression models
246
confirmed the importance of the use of environmental variables to characterize the snail
habitat in the endemic area of the state of Minas Gerais. The methodology used in this
study can be utilized to control schistosomiasis. It is recommended the use of GPS for
field surveys together and the application of this methodology with images of better
spatial resolution (10-30m) in other states for validation. Financial Support: Fiocruz, CNPq
(302966/2009-9, 490336/2007-8, 380203/2004-9, 308253/2008-6).
247
EvaLUatE tHE EffECt Of agE In REdUCIng tHE PREvaLEnCE Of SCHIStOSOMIaSIS,
USIng COHORt anaLYSIS
Erica Magueta; Michele Mendonça; alessandra Miranda; Rosiane araújo; alda Maria Soares
Silveira; Elaine Speziali; Elizabeth Castro Moreno; Lucia alves de Oliveira fraga
UNIVALE- Universidade Vale do Rio Doce, MG - Brasil
Introduction: This study aimed to estimate the current prevalence of schistosomiasis and
characterize the population of two endemic areas close Governador Valadares municipality,
Minas Gerais state. Methodology: Surveys were used, addressing individual and household
records to identify possible risk factors associated with the disease. The presence and number
of parasite eggs per gram of feces were determined by the method Kato-Katz. Results: Study
participants were 117 families with 449 individuals and 382 fecal samples were analyzed.
The prevalence of schistosomiasis in C. Bernardo was estimated at 12.5% (95% CI 8.4 to
17.8%) and Melquiades 31.9% (25.3 to 38.5%), indicating a higher prevalence of disease in
Melquiades. The prevalence ratio for C. Bernardo was 1:7 indicating that for every one person
infected with S. mansoni 7 persons were without infection. In Melquiades, the prevalence
ratio was 1:2, indicating a greater chance of infection by the disease in this location. Infected
individuals had a low parasite load seen in these locations, where the count of eggs per gram
(epg) of feces was less than 100 epg in most cases (CB = 36 and MQ 3Q = 92), however, the
parasite load in Melquiades was significantly higher when compared to participants in C.
Bernardo (p <0.005). The detailed survey of the historical series of secondary data from the
rural districts, in the period of 1990-2009 (Bernardo) and 1997-2009 (Melquiades) showed
that the 117 families who participated in the study in 2009, ninety- two (92) had participated
in previous studies done by our laboratory. In the nineties (1990/1997) the prevalence of
schistosomiasis in both districts was approximately 60%. Conclusions: All individuals who
participated in the epidemiological survey in 2009 were invited to participate of clinical
evaluation, anthropometric measurements, blood pressure and ultrasound examination.
Univariate and multivariate analysis of all data collected are being made.
248
EvaLUatIOn Of tHE SCHIStOSOMIaSIS SURvEILLanCE SYStEM In aLagOaS StatE,
BRazIL, fROM 2004 tO 2008.
gilmara Lima nascimento; Maria José Menezes2; Jean Lúcia dos Santos; Wildo navegantes
de araujo
Ministério da Saúde/SVS, DF - Brasil
Introduction: The Schistosomiasis mansoni (SM) remains an important health challenge in
Brazil, with a mortality rate of 0,3/100.000 inhabitants in 2007. In Alagoas state, located
in Northeast region of Brazil, the transmission is recognized in 70% of municipalities and
the percentage of Schistosoma. mansoni carriers documented in 2007 was 8.4%. The
intermediate host is documented in 40% of Alagoas municipalities currently classified as
non-endemic. The evaluation of surveillance systems aims to ensure adequate and efficient
monitoring of relevant health problems. The objective was to evaluate the schistosomiasis
surveillance system (SSS) in Alagoas from 2004 to 2008. Methodology: Following the Updated
Guidelines for Evaluating Public Health Surveillance Systems of the CDC (2001) we conducted
the assessment including the description of the system, its attributes and utility based on the
system’s stated objectives. Data sources were the databases of the Schistosomiais Control
Programme Information System (SISPCE); the reportable diseases information system (Sinan)
for schistosomiasis, the Mortality Information System (SIM), standardized questionnaires
administered to 18 key respondents working with surveillance, control and health care at
the State Health Secretariat and Municipal Health Secretariats of: Maceió which is a endemic
city where a reference hospital for severe SM cases is located, São Miguel dos Campos, an
endemic city, and Major Isidoro (MI), a city on the border of the endemic area and classified
as non-endemic. We interviewed six people from MI who had positive coproscopy in 2006.
Results: In the endemic area the surveillance is based on active case finding. Stool surveys
are performed and registered in SISPCE. In the non endemic area (32 cities) surveillance
is passive and notification to Sinan is mandatory. Concerning flexibility, two major changes
were reported in 2006: notification of cases passively found by primary health care units to
SISPCE and therefore 500 cases from such institutions are entered in the database. The other
change was the recommendation to record severe cases in Sinan, however such cases were
not encountered in this database. The case definition in place requires the presence of S.
mansoni ova in feces or tissues and this information was not available for the majority of the
severe cases admitted at reference hospitals. Regarding data quality, SISPCE database does
not include identification variables therefore it was not possible to evaluate duplication. The
completeness of the essential variables in SISPCE was >90% and Sinan completeness ranged
from 36 to 86%. The SISPCE inconsistencies were <20%, however 92% of Sinan records
consisted of non-severe cases from endemic cities, which were supposed to be recorded in
SISPCE and not in Sinan. Acceptability was considered low, 24 to 28% of the population who
received stools containers did not provide a specimen. The percentage of S. mansoni carriers
not treated after the surveys ranged from 14 to 17%. No evidence was found of systematic
monitoring of non-endemic areas where the presence of intermediate hosts is documented.
The 140 cases (mortality rate: 2.2/100.000 in 2007) of death from SM recorded in the SIM in
2007 and 2008 were not detected by Sinan in the study period. The delay to closure of the
investigation in Sinan was >30 days for 62% of the notifications in 2008 and in the period of
249
the study 19 cities currently classified as non-endemic reported autochthonous cases of SM
and only one field investigation of one site was conducted. Concerning representativeness
of SISPCE 57% of the S. mansoni carriers were male, 70% under the age of 30 years. In 2010
we interviewed 6/26 S. mansoni carriers diagnosed and treated in 2006 and 4/6 reported
that they have not been outside MI. All interviewed people have contact with Açude da
Nação, major water source of their neighborhood. Therefore 4/6 S.mansoni carriers could be
autochthonous. Study limitations include use of a convenience sample of cities. Conclusions:
The SM surveillance system in Alagoas was considered complex, flexible, had good quality
data in SISPCE, but limited data quality in Sinan. The acceptability was low as well as
timeliness of investigation of vulnerable areas. The sensibility was low to detect deaths
related to SM and we did not find evidences of meeting the stated objectives of preventing
the expansion of the transmission area. Recommendations: Review the case definition
of the passive system allowing notification of severe cases without positive coproscopy,
include identification variables in SISPCE to allow linkage with the hospital morbidity and
mortality databases, and implement surveillance activities in non-endemic areas where
the intermediate host is present. Finally the possible expansion of the SM transmission in
Alagoas should be investigated.
250
faCtORS aSSOCIatEd WItH MaIntEnanCE Of SCHIStOSOMIaSIS In tHE fORESt zOnE
Of PERnaMBUCO
verônica Santos Barbosa; Karina Conceição araújo; tadeu Rodrigues; Constança Simões
Barbosa
Introduction: The schistosomiasis remains at endemic levels in the rural municipalities of
the forest zone of Pernambuco (FZP). The transmission of disease is determined by biological
factors. However, the production and maintenance of endemic process are influenced by
political, sociocultural and sanitary environmental factors, being essential knowledge and
understanding of these factors, to avoid and control this disease. The mapeament of diseases
with their geographical pattern of occurrence is essential for monitoring the actions of Public
Health Surveillance. The tool that has been used for knowledge of the spatial and temporal
factors associated to schistosomiasis is Geographic Information Systems (GIS). The GIS allows
you to assemble a cartographic base to cover the many determinants of the disease, giving a
holistic view on the state of health of a population. The objective of this study was verified the
socioeconomic factors and sanitary conditions associated with the occurrence of schistosomiasis
in the FZP. Methodology: The study was conducted using secondary data obtained in the
Schistosomiasis Laboratory of CPqAM/Fiocruz, about the prevalence and intensity of infection for
schistosomiasis. The independent variables (houses with: water supply, treated water, sewage,
garbage system, masonry and electricity) were collected in the Information System of Basic
Attention (SIAB / DATASUS). To test the association between variables, we used a generalized
linear model, logistic and gamma, being used the AIC criterion to select the best model. The spatial
analysis of data was realized using the TerraView program. Results: Amongst the six independent
variables applied to the model, only the presence of houses with water supply by the public
system and houses with treated water showed significant and negative statistical association with
the prevalence of schistosomiasis in the municipalities of FZP. The model also showed higher
coefficient for FZP south towards north. For the intensity of infection, the model indicated a
negative association for the variables houses with water supply and houses with sewage, but
only the first variable showed significant association. For the intensity of infection the model also
showed that the south ZM has a higher coefficient that the north. Most municipalities in the FZP
have 50.1 to 70% of houses with coverage of water supply and the spatial distribution of these
municipalities is similar between zones north and south. With regard to water treatment most
municipalities also presents coverage from 50.1 to 70% and their distribution is higher in the
center. The predominant covering of sewage is less than 20% and the greater concentration it’s in
the north FZP municipalities. Conclusions: In the FZP the presence of houses with water supply
and with treated water in the municipalities reduces the prevalence of schistosomiasis. The
prevalence decreases also in the municipalities of the south FZP over north, so this latter has a
higher risk of infection by the disease. The intensity of infection decreases with the increase in the
proportion of houses with water supply and sewage. In addition, municipalities in southern had
higher intensities of infection than the northern. The spatial distribution of municipalities with
respect to water supply is similar between the north and south FZP, the spatial distribution refers
to houses with treated water concentrated in the center and in relation to sewage predominates
in north FZP.
251
gEOSPatIaL anaLYSIS On HUMan CaSES Of ManSOn’S SCHIStOSOMIaSIS In a
HORtICULtURaL COMMUnItY In tHE zOna da Mata, PERnaMBUCO, BRazIL.
Onicio Batista Leal neto; Elainne Christine de Souza gomes; fabrício andrade Martins Esteves;
thiago Yury Cavalcanti galvão; Karina Conceição o araújo; Constança Simões Barbosa
Introduction: Schistosomiasis is an endemic disease in the Zona da Mata, Pernambuco,
and it continues to expand its area of incidence because of environmental conditions
and/or human actions that favor proliferation of the vector molluscs and maintenance
of the Schistosoma mansoni cycle. In rural communities in Pernambuco, absence
of sanitation together with lack of investment in health information and education
contribute towards fecal contamination of freshwater catchment areas that serve
these communities. Investigation of factors within the environmental space in which
the disease occurs is of fundamental importance, in providing support for intervention
strategies with consequent improvement in the local quality of life. The aims of this
study were: (1) to ascertain the prevalence of schistosomiasis in the community; (2) to
determine the parasite load in human cases; (3) to perform malacological recognition (4)
to spatially locate disease cases and vector breeding sites; and (5) to construct thematic
maps to express the local epidemiological landscape. Methodology: Between November
2008 and March 2009, a coprological survey was conducted among 310 individuals in the
community of Natuba, in the municipality of Vitória de Santo Antão, Pernambuco, using
the qualitative methods of spontaneous sedimentation and floatation/centrifugation
in 33% zinc sulfate and the Kato-Katz quantitative technique for counting Schistosoma
mansoni eggs. During this period, an active search for snails of the genus Biomphalaria
was carried out in natural aquatic environments and in market garden irrigation ditches,
and some specimens were dissected for species identification. The positive human cases
of S. mansoni and the breeding sites of the vector snails were georeferenced by means of
the Global Positioning System (GPS). The database was constructed using the TerraView®
software. Kernel estimators were generated for the human cases and the parasite load
was demonstrated by means of chromatic gradients formed in the regions where the
cases were concentrated. Results: Parasitosis was found to occur in 61.3% of the study
population, and the high prevalence of Schistosoma mansoni stood out (28.4%). The
parasite load among the parasitized individuals was categorized into three bands of
infection intensity: 0 to 99 opg (low intensity); 100 to 399 (medium intensity); and >
400 (high intensity). Among all the positive human cases, 83.91% were of low intensity,
14.94% medium intensity and 1.15% high intensity. Among all the aquatic environments
investigated, vector snails were only found in irrigation ditches, and were identified as B.
straminea and not infected with S. mansoni. The georeferenced and spatially organized
maps showed the distribution of schistosomiasis cases and the concentration of infection
intensity in the community. Allied with this information, the observations on the local
microenvironment made it possible to conclude that the individuals had become
infected through laboring practices, given that in the market gardens, systematic human
exposure to the vector snails that were brought into the gardens through irrigation was
observed. Conclusions: The thematic maps show greater case concentration and greater
252
numbers of individuals with high parasite loads in the eastern region of the community,
which represents a risk of disease expansion to new environments, given that the water
bodies that form natural breeding sites for Biomphalaria are located there. This setup
points towards a future scenario of expansion of schistosomiasis into neighboring areas,
thus putting at risk another segment of the population that does not comprise farm
workers.
253
gEOSPatIaL dIStRIBUtIOn Of vECtOR MOLLUSCS fOR SCHIStOSOMIaSIS On tHE
COaSt Of PERnaMBUCO and tHE MEtROPOLItan REgIOn Of RECIfE
Constança Simões Barbosa; Onicio Batista Leal neto; Manuel amarista Sevilha; fabio Melo;
Jones albuquerque; Elainne Christine Souza gomes; Reinaldo Souza dos Santos
Introduction: The expansion of schistosomiasis to disease-free areas on the coast of
Pernambuco has been accompanying the process of disorderly exploitation of natural
resources for tourism purposes, through construction of vacation homes and hotel
infrastructure. Unemployed multi-parasitized populations are attracted by the offer of casual
jobs in these areas and congregate in marginal zones where there is no access to potable
water and no basic sanitation system. The way in which these spaces are occupied, allied
with environmental and climatic conditions that favor the introduction and maintenance of
vector snails for schistosomiasis, have created a perfect epidemiological landscape for the
installation and reproduction of new disease foci. Methodology: Between 1998 and 2010,
foci of the vector mollusc Biomphalaria glabrata were identified and monitored in coastal
localities (Table 1) where this species has been shown to predominate. From 2005 onwards,
the presence of the species B. straminea started to be detected in coastal environments.
So far, four new breeding sites and transmission foci have been recorded (Table 2). In 2006,
for the first time, both species were found in the same locality, Forte Orange beach, on
the island of Itamaracá. In 2007, a new focus of B. straminea was detected at Janga beach,
municipality of Paulista. These findings suggest that this mollusc species (B. straminea), which
is characteristic of the rural endemic area in Pernambuco (Zona da Mata), is being introduced
and is adapting to coast localities in this State. In 2008, a scientific expedition organized by
specialists in the ecology and epidemiology of schistosomiasis updated the information on
occurrences of these coastal foci in all the water bodies found in a strip of up to 1 km in
width, going inland from the coast. In the laboratory, the molluscs that did not eliminate
cercariae of S. mansoni when exposed to light were subjected to specific molecular biology
techniques (nested PCR), to detect the DNA of the parasite. This expedition also detected the
presence of B. straminea parasitized by S. mansoni in the Apipucos Lagoon, which is located
in an upmarket district of Recife. Results: Figure 1 shows the updated geographic distribution
of the foci of vector molluscs of schistosomiasis on the coast of Pernambuco and the spatial
analysis on the risk of transmission of this disease in localities such as: Carne de Vaca beach
and Pedra point (Goiana); Forte and Enseada dos Golfinhos beaches (Itamaracá); Janga
and Pau Amarelo beaches (Paulista); Náutico Lagoon (Jaboatão); Enseada beach (Cabo);
Porto de Galinhas beach (Ipojuca); Tamandaré beach (Tamandaré); and Apipucos Lagoon.
Conclusions: These coastal foci are generating acute human cases of schistosomiasis, such
that there is a need for intervention by the State, in partnership with private enterprise, to
implement basic sanitation actions and/or environmental actions in these localities, in order
to ensure sustainability of the environment, visitors’ wellbeing and improvement in the local
population’s quality of life.
254
HIv InfECtIOn and EndEMIC dISEaSES: CaSES‘S dIStRIBUtIOn Of aIdS and
SCHIStOSOMIaSIS In aLagOaS
arthur Maia Paiva; Jean Lúcia dos Santos
Hospital Universitário-UFAL / SESAU, AL - Brasil
Introduction: In Brazil, it is estimated that approximately 3 million people are parasitized by
Schistosoma mansoni and the region with the highest prevalence is the Northeast, where
Alagoas stands out, along with Pernambuco, Sergipe and Bahia, as the State hyperendemic for
the disease. Moreover, with the growth of the AIDS epidemic and its trends to internalization
and impoverishment, we expect an increase in the prevalence of coinfection between HIV
and endemic diseases, which in our environment include schistosomiasis. Methodology:
We reviewed medical records of one case of coinfection HIV / schistosomiasis reported by
University Hospital - UFAL in 2010, selected for reporting. Data on schistosomiasis, updated
to 06/21/2010, were obtained from the Information System of the Schistosomiasis Control
Program of Department of State Health of Alagoas (SISPCE / DIVEP / SESAU). Data on AIDS
cases, tabulated by year of diagnosis according to the municipality of residence, consolidated
on 23.05.2010, were obtained from the Information System for Notifiable Diseases (SINAN)
for the period 1986 to 2000 and through his NET version (SINAN NET) for the period 2001
to May 2010. Results: Case 1 - JASF, male, 38 years, Alagoas, living in Maceió, 1 month ago
with cough, evening fever, night sweats, weight loss, anorexia, diarrhea, impaired general
condition, palpable spleen at the umbilicus, liver 5 cm below the xiphoid process; Rx thorax:
partial opacification of left hemithorax, with a slight deviation to the right mesdiatino;
rapid test anti-HIV reagent, prompted by the probable tuberculosis with pleural effusion,
negative AFB in sputum, total protein 7 mg / dl, albumin 2.7 mg / dl, SGOT: 58 SGPT: 57; CBC:
leukocytes/mm3 2270, with 179 lymphocytes, 324 monocytes, 140 eosinophils: 10% rods,
hematocrit: 28%, hemoglobin : 9.04 g / dl, 120,000 platelets/mm3; negative serology for
hepatitis B and C, HTLV, Chagas, leishmania human, syphilis. stool examination: hookworms.
Abdominal ultrasonography total: “Liver of normal contours and topography, an increase in its
size at the expense of the left lobe and modifying their texture by increasing the echogenicity
of vessel walls and portals diffuse increase in echogenicity of the parenchyma without signs
of expansive processes; with increased portal vein diameter, 1.8 cm; complementation with
Doppler collor study showed liver fugal flow; splenic vein at the topography of the splenic
hilum with a caliber of 1.35 cm (NR <1.2 cm). Conclusion: signs of diffuse parenchymal
disease with hepatomegaly and at the expense of the left lobe, marked splenomegaly, signs
of collateral circulation “. It was begun tuberculostatics, trimethoprim-sulfamethoxazole
prophylaxis, albendazole, praziquantel, after HAART, with good response. After 6 weeks using
HAART: CD4=44 cells/mm3; viral load <50 copies/ml. Of the 102 municipalities in Alagoas,
69% (70/102) comprise the endemic area of schistosomiasis, with a concentration of severe
cases and mortality attributed to the disease, approximately one million people living under
the risk of infection in this area. Comparative data for the last five years have shown through
surveys conducted coproscopic in the endemic area, maintenance of prevalence around 8010% (8.76% in 2009), with 24.29% of the municipalities presenting prevalence greater than
10%. Among subjects who were positive on parasitological examination with high parasite
loads (up to 17 eggs / slide), 93% were in the age groups 5-49 years. From 1986 until June
255
2009 were reported in SINAN 2738 AIDS cases in Alagoas, with an annual average of 211
cases over the last five years. Among the ten cities with the highest number of AIDS cases,
nine are located in an endemic area for schistosomiasis and account for 81.22% (2224/2738)
of AIDS cases reported in the state. Currently only 14 (13.72%) of the 102 municipalities are
silent as to the notification of AIDS cases. Among the 70 municipalities in the endemic area
for schistosomiasis, only 4 (5.71%) are still silent about the reporting of AIDS, ie 94.29%
of the 70 municipalities of the endemic area of schistosomiasis cases are affected by AIDS
and are responsible for 90.50% (2478) of all cases of the syndrome reported in Alagoas.
Conclusions: The coinfection between S. mansoni and HIV can influence many aspects of
both infections. Of schistosomiasis, for example, we know that there may be decrease in
the number of eggs eliminated in feces and increased susceptibility to reinfection by S.
mansoni. Due to the expansion of the AIDS epidemic, the existing data in our state indicate
that the differential diagnosis of this co-infection should always be considered in patients
with HIV / AIDS that show positive epidemiology for schistosomiasis and / or any events
that may be associated with it or its complications. It is possible that the prevalence of HIV/
schistosomiasis coinfection is being underestimated.
256
HOSPItaLIzatIOn and MORtaLItY Of SCHIStOSOMIaSIS ManSOnI In CEaRa StatE,
BRazIL, fROM 2000 tO 2009.
vivian da Silva gomes; Ricristhi gonçalves de aguiar gomes; alberto novaes Ramos Junior;
Carlos Henrique Morais de alencar; Jorg Heukelbach; fernando Schemelzer Moraes Bezerra
Secretaria Estadual de Saúde, CE - Brasil
Introduction: The distribution of schistosomiasis in Brazil is characterized by endemic areas
on the coast and forests of some states in the Northeast, especially Alagoas, Sergipe, Bahia,
Ceará and Northern parts of Minas Gerais state. Mortality due to Schistosomiasis mansoni
in Ceará State was very high in the 1970s, with most cases in the chronic hepato-splenic
form. This led to the creation of a task force realizing extensive mass treatment. Since
1984, selective mass treatment was realized after faecal examination, in districts with a
prevalence >20%. This resulted in reduction of morbidity and mortality. Here we describe
hospitalization and number of deaths caused by Schistosomiasis mansoni in Ceará between
2000 and 2009. Methodology: We analyzed secondary data from 2000-2009, provided by
national data sets: information system on hospitalization (SIH), and information system on
mortality (SIM, DATASUS). Data on hospitalization and mortality due to schistosomiasis were
analyzed. Results: In total, 150 hospitalizations occurred due to schistosomiasis in Ceará in
the study period. The hospitalization rate decreased from 0.084/100.000/year in 2000 to
0.026/100.000/year in 2009. The distribution by age shows that the rate of hospitalization
due to schistosomiasis is more frequent in the age group of 20-49 years. In total, 150 deaths
occurred, resulting in an overall mortality rate of 0.006/100.000/year. Conclusions: In
endemic areas, mortality rates can be used as an indicator to monitor intervention measures.
Morbidity and mortality due to schistosomiasis decreased significantly in the last decade,
but control efforts need to be maintained.
257
IdEntIfICatIOn Of MaIn EntEROPaRaSItOSIS and SCHIStOSOMIaSIS ManSOnI
In StUdEntS Of tWO SCHOOLS PERIPHERaL Of Santana Of IPanEMa, StatE Of
aLagOaS, BRazIL.
Márcio Bezerra Santos; Erlon Oliveira dos Santos; Karina Conceição gomes Machado de
araújo
Introduction: Schistosomiasis mansoni is a serious parasitic disease, of evolution chronic,
found in Brazil excellent conditions for development and expansion, while intestinal parasites
deserve special mention when referring to the contamination of children, especially in the
age group ranging from 5 to 12 years of age, the initial phase of school life. The prevalence of
both diseases is undoubtedly high in developing countries, as well as in Brazil, its distribution
is presented in different ways, depending on the region of each country and directly from
the sanitation conditions of socioeconomic status and hygiene habits individuals. Municipal
schools outlying town of Santana of Ipanema-AL, are located in areas where the conditions
of infrastructure are inadequate to the survival of its inhabitants. This situation is, therefore,
students in this region vulnerable to infection by intestinal parasites. This study aimed to
identify the incidence and risk factors of intestinal parasites and schistosomiasis among
school children of municipal schools located in the peripheral area of the municipality.
Methodology: Participated in this study, students from kindergarten and 1st to 5th year of
elementary school two municipal schools of the peripheral region of the city. This research
consisted of a descriptive and analytical epidemiological study, a cross-sectional, conducted
during March-July 2010. 107 students participated, randomly selected, which represented
56.8% of students from two schools. To obtain data, a questionnaire was applied in
investigative time the student was in their respective school. To collect the feces collectors
were distributed previously identified with name, number and numbering of the student.
Analysis of samples were performed by the methods of Hoffman, Pons & Janer (1934) and
Kato-Katz (1972). Results: Among the total of students who carried out tests 69.51% were
positive for some type of parasite or commensal. Among those identified, protozoa Giardia
lamblia (27.96%) and the complex Entamoeba histolytica/dispar (8.6%) were the most
present. Among the helminths were identified Ascaris lumbricoides (8.6%) Ancylostomidae
(4.3%) and Trichuris trichiura (4.3%). Among the commensal, the protozoa Endolimax nana
(19.36%) and Entamoeba coli (18.29%) stood out. It was not possible to identify the infection
by Schistosoma mansoni among the students. The frequency of monoparasitism (59%) was
higher in relation to multiple parasitic infections (41%). The Kato-katz was more sensitive in
determining the presence of helminth eggs in feces, with a percentage of 20.74% positivity,
almost twice the value expressed by HPH (10.98%). Students who use untreated water have
an infection rate (66.70%). Conclusions: The occurrence of intestinal parasites in students
of municipal schools is high. Environmental and behavioral aspects of school are important
in the transmission of these diseases, emphasizing the importance of investments in public
health programs in order to minimize the prevalence and risk of infection by intestinal
parasites.
258
LOngItUdInaL StUdY On SCHIStOSOMIaSIS and SOCIOECOnOMIC vaRIaBLES In a
RURaL aREa In MInaS gERaIS StatE, BRazIL.
Humberto ferreira de Oliveira Quites; Helmut Kloos; Leonardo ferreira Matoso; Mery natali
Silva abreu; João Paulo amaral Haddad; Rodrigo Corrêa-Oliveira; andréa gazzinelli
Escola de Enfermagem-Universidade Federal de Minas Gerais, MG - Brasil
Introduction: Schistosomiasis is strongly associated with socioeconomic factors worldwide. With socioeconomic development and improvements of health services in rural
areas over the last decade in Brazil, schistosomiasis risk and infection rates have declined
in most areas. The objective of this study was to identify the socioeconomic determinants
of schistosomiasis in Virgim das Gracas rural area in Minas Gerais State, Brazil, during the
period 2001-2009. Methodology: The study population comprised 575 persons in 2001 and
553 in 2009, including 377 individuals who lived in Virgem das Graças at these two points in
time. Socioeconomic and water contact data were collected during household surveys using
a questionnaire. All participants were examined parasitologically in 2001, 2002, 2005 and
2009 using the Kato Katz method and treated with praziquantel. Results: The results showed
a reduction in prevalence from 58.3% to 26.8% and in geometric mean egg counts from
58.93 (CI95% 56.76-61.11) in 2001 to 13.45 (CI95% 11.29-15.60) in 2009. The number of
houses using the streams decreased during those 8 years, largely as a result of increased use
of water containers, piped water and electric water pumps. In bivariate analysis, a significant
relationship was found between schistosomiasis prevalence and household income, contact
with potentially safe water, ownership of motorized vehicles, latrines and showers (p<0.01).
In the multivariate logistic regression model S. mansoni infection in 2009 was significantly
correlated with age group 10-19 years (OR=1.16 CI95% 1.02-1.31) and absence of showers
(OR=1.19 CI95% 1.08-1.30). Self evaluation of family health conditions showed that 30.0% of
all households attributed health improvements during the study period to increased income
from the government’s “Bolsa Familia” social program, which benefits poor families and was
implemented in 2004. Conclusions: The finding that improved socioeconomic conditions
between 2001 and 2009 were associated with increased use of a safer water supply and
declines in S. mansoni infection warrants further studies in other communities. FINANCIAL
SUPPORT: CNPQ, FAPEMIG, NIH-ICIDR Grant (1R03AI071057-01) and CAPES.
259
MOLLUSCICIdaL aCtIvItY Of HExanE and dICHLOROMEtHanE fRaCtIOnS Of
EtHanOL ExtRaCt Of gIngER (ZInGIBER OFFICInAlE) On tHE BIOMPHAlARIA
GlABRATA, IntERMEdIatE HOSt Of SCHISTOSOMA MAnSOnI
teiliane Rodrigues Carneiro; antonio gomes da Silva neto; James almada da Silva; ana Lúcia
de Paula Hanemann; Paulo Cesar vieira; Renata de Sousa alves; fernando Schemelzer de
Moraes Bezerra;
Introduction: Research in natural products areas are very promising in the search for
new drugs in different fields. One of this, is the control of mollusc intermediate hosts of
schistosomiasis, a parasitic disease caused by Schistosoma mansoni, which currently affects,
according to World Health Organization, 200 million people in tropical and subtropical areas
of the world. Population control of the gastropod mollusc Biomphalaria glabrata has been
done with synthetic molluscicides that are expensive and cause ecological imbalance. The
plants contain a number of toxic components and / or antinutritional factors of different
nature, which can be used in a beneficial way, as in the control of the snail Biomphalaria
glabrata, for example. The biochemical and biological studies of these compounds
contribute to the use of such constituents in several areas: agriculture, pharmacology,
medicine, immunology, toxicology, among others. Ginger (Zingiber officinale) is a plant
used as flavoring and as a medicinal herb since ancient times by people from the east.
Presents in its chemical composition, volatile compounds (terpenes) and non-volatile
(phenolics and alkaloids). Monoterpenes and sesquiterpenes are found among the volatile
constituents. Many of these volatile compounds contribute to the distinctive aroma and
flavor of ginger. In the class of non-volatile compounds are present gingerois, shogaois
is stopped, mostly. Gingerois are phenolic compounds that represent a homologous
series of ketones, that is, show differences in the alkyl chain length. In search of new
molluscicides from plant origin, we analyzed the activity of two fractions of ethanol extract
of ginger against the snail Biomphalaria glabrata. A fraction has the sesquiterpenes as
major constituents (hexane) and another (dichloromethane) gingerois, the latter products
already have several other biological / biochemical activities. Methodology: The used
snails were Biomphalaria glabrata (Say, 1818) species obtained from the strain maintained
in the Research Laboratory of Parasitology and Biology of Molluscs DACT / FFOE / UFC.
The ginger extract was obtained by rhizome maceration using ethanol 99.5%. This extract
was subsequently subjected to a fractionation process (liquid-liquid partition) to obtain
the hexane (sesquiterpenes) and dichloromethane (gingerois). In the immersion test of
the molluscicidal activity on Biomphalaria glabrata, we used 40 mL of solutions (positive
control, and negative samples) per snail. The containers containing 10 young snails of
uniform size (10-13 mm) for each sample in duplicate, were covered with gauze and kept
at room temperature for two days. After the period of exposure to the samples, the snails
were washed with distilled water and kept in new containers with the same initial volume,
but with distilled water containing lettuce ad libidum for a period of 48 h. The count of
surviving snails was performed throughout the experiment (four days). Were tested 5mg/
ml and 10mg/ml concentrations. The agents used to solubilize the fractions to be tested
were DMSO, TWEEN. Results: The fraction of the dichloromethane extract of ginger at
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10 mg / ml concentration had 100% mortality with 24 hours of exposure and 5mg/ml
concentration showed 20% mortality within 48h of exposure. The hexane extract at the
tested concentrations and the observed period was not toxic to the snails. Conclusions:
Dichloromethane fraction showed molluscicidal activity at the highest concentration and
shorter evaluated time (100% mortality). However, in order to better evaluation of the
molluscicidal activity of this fraction, subsequently, other concentrations should be tested
to determine the LD50 and LD90 subfractions and purified from this fraction and also
should be evaluated for the search of the specific compounds responsible for this activity.
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MOLLUSCICIdE aCtIvItY Of tHE EtHanOL ExtRaCt Of tHE LEaf Of AnACARDIuM
OCCIDEnTAlE L. aBOUt BIOMPHAlARIA GlABRATA (SaY, 1818), IntERMEdIatE HOSt
Of SCHISTOSOMA MAnSOnI (SaMBOn, 1907).
teiliane Rodrigues Carneiro; Morgana M. O. Barboza; Márcia M. M. Marques; ana R. a. Silva;
Sérvio Q. Júnior; fernando Schemelzer de Moraes Bezerra; Maria I. f. guedes
Introduction: Schistosomiasis is a human parasitic disease widely distributed, mainly in
tropical and subtropical areas of the world, affecting 200 million people, while about 500
million to 600 million are at risk of being infected. In Brazil this disease occurs from the
northeast to the south center of the country and affects about 8 to 12 million people. In
areas where the disease is highly endemic, molluscicides are used as a prevention strategy,
however the high cost of synthetic molluscicides, concern about its toxic effects on non-target
organism and a possible report of resistance among the mollusks, require the development
of safer and less costly. In contrast, the use of native plants with molluscicidal activity may
represent a cheap alternative, and not pollute the environment. Molluscicides from plant
origin have been studied on the feasibility and safety since the 1930s. Molluscicides have
been studied as about 1,100 plant species. Products originating from various parts of plants
such as bark, leaves, roots, have shown molluscicidal action. Despite the increase in this area
of study, available data show that only 15-17% of plants were studied for their potential.
The aim of this study was to evaluate the molluscicidal activity of the ethanol extract of
leaf of Anacardium occidentale, belongs to the family Anacardiaceae, against Biomphalaria
glabrata, intermediate host of S. mansoni. The leaves of cashew were collected on private
property in Fortaleza, Ceara, and dried at 50 ° C. Methodology: To obtain the ethanol extract,
leaves were mechanically crushed and subjected to extraction with absolute ethanol at room
temperature for one week. The resulting solution was filtered and then concentrated on
rotary evaporator. Aliquots of stock solution were added to dechlorinated water to obtain
concentrations of 100, 10 and 1 ppm. Testing for molluscicidal activity were performed
according to the standards of the World Health Organization, 10 adult snails were used per
group, with between 10 to 13mm. The snails were placed in 200 ml of test solution at the
concentrations described above. Forty-eight hours later, the animals were removed, washed
three times with distilled water and transferred to another tank where they remained for a
period of recovery with time equal to the exposure, when mortality was recorded. Results:
The ethanol extract of leaf of A. occidentale showed 100% mortality at a concentration of
100 ppm with 48 hours of exposure. Conclusions: We conclude that the ethanol extract of A.
occidentale have molluscicidal activity against B. glabrata at levels 100 ppm/48hs, however,
indicated a continuation of tests with various concentrations and periods.
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MORPHOLOgICaL CHangES In SCHISTOSOMA MAnSOnI adULt WORMS RECOvEREd
fROM nItRIC OxIdE SYntHaSE dEfICIEnt MICE fEd a LOW-PROtEIn dIEt
Renata Heisler neves; Renata Pinto Ramos; Silvia M. L. Montenegro; Eridan Medeiros
Coutinho; José Roberto Machado-Silva
Instituto Oswaldo Cruz – Fiocruz, RJ - Brasil
Introduction: In endemic zones for Schistosoma mansoni infection, an overlapping of
undernutrition and parasite infection is frequently observed. Malnutrition has been
considered as a factor able to modify the host-parasite-environment system, aggravating
the course of schistosomiasis by breaking the equilibrium in the relationships among the
components of this system. Nitric oxide (NO), a biological molecular mediator produced
in mammalian cells by enzymatic oxidation of L-arginine mediated by NO synthase, is
relevant as an intra and inter-cellular messenger and as a cytotoxin released during
various physio-pathologic events, including immunological reactions and inflammation.
The inducible NO synthase (iNOS) is one of the three NOS isoforms present in virtually all
cells and expressed in response to pro-inflammatory cytokines (IFN-γ, TNF-α, IL-1β) and/or
microbial products (lipopolysaccharides). In well-nourished mice, NO production increases
following the deposition of schistosome eggs in the liver. Overproduction of NO can induce
tissue damage and may contribute to morbidity during schistosome infection. This study
presents preliminary results on morphological features of Schistosoma mansoni adult
worms recovered from nitric oxide syntase knockout mice (KOiNOS) fed a low-protein diet.
Methodology: Male 21-day old C57BL/6 mice, deficient in iNOS (KOiNOS) were used. Mice
were obtained from Fiocruz (Rio de Janeiro) and maintained in the CPqAM animal facilities.
All the procedures involving mice were approved by ethical committee for the use of animals
(CEUA/Fiocruz, P0201/03). A strain of S. mansoni isolated from Belo Horizonte (BH) and
kept in the laboratory was used. Mice were infected percutaneously with 30 cercariae shed
from Biomphalaria glabrata. Malnutrition was induced in mice by ingestion of a regional
basic diet (RBD), based on the dietary standards of human populations living in areas
where schistosomiasis is endemic in Northeast Brazil adapted to the requirements of the
laboratory mouse. The KOiNOS group was subdivided into infected malnourished (n = 10),
and infected well-nourished mice (KOiNOS IE) (n = 10). After 90 days of infection, all mice
were anaesthetized and sacrificed. Adult worms were recovered and fixed in AFA (alcohol,
formalin and glacial acetic acid) at room temperature. A sample of recovered worms of both
gender was stained with 2.5% hydrochloric carmine, dehydrated in a series of alcohols (70%,
90% and absolute), clarified in methyl salicylate with Canadian balsam (1:2) and preserved as
whole-mounts on glass slides. Confocal images were captured with a LSM 510–ZETA confocal
laser microscope (Zeiss, Germany), under reflected mode with a 543nm He/Ne laser and LP
570 filter. Morphological changes in the tegument, suckers and reproductive organs of male
and female adult worms were analyzed. Results: Females: Confocal images from the anterior
end of female worms from the malnourished group (KOiNOS IM) showed that the oral and
ventral suckers were normal in appearance. The follicular vitelline glands were distributed
into two lateral bands, in which few vitelline cells were present with undefined nucleus.
Ovary presented cellular differentiation but some degenerated oocytes were seen. Towards
the anterior end, the common vitelline duct joins the oviduct, which expands slightly to
263
form the ootype, comprising flattened cells and unclear nuclei, where a developing embryo
enclosed by its shell was observed. The ootype opens into a long uterus with no evidence
of eggs. Control well-nourished group also showed well developed suckers. At the posterior
end vitelline glands with few remaining follicles were detected. In contrast, well-fed female
worms presented an elongated ovary filled with normal mature oocytes. Ootype presented
normal features and always contained developing eggs. In both groups, a light superficial
desquamation of the tegument was found. Males: Both groups (malnourished and wellnourished) had testicular lobes with fewer differentiated germinal cells and vacuolated areas.
In the seminal vesicle there were a few or none spermatozoa. At the anterior end of males
both suckers and gynaecophoric canal were covered with small spines irregularly arranged.
Most structural damages were evidenced in the tegument of worms from hosts with a poor
nutritional status. In nitric oxide knockout mice, the worms were partially disintegrated,
had rudimentary tubercles, few spines and tegument with roughened appearance. The
tegumental layer was destroyed and vacuolated areas were detected in the subtegumental
region. Male adult worms showed a flacid musculature and a feeble degree of muscularization
on the dorsal surface. Conclusions: Adult male and female S. mansoni recovered from nitric
oxide synthase knockout mice developed several morphological changes in the tegument
and internal structures, but no relationship could be detected between these anatomical
changes and the nutritional status of the host.
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MORPHOLOgY Of “BREvIfURCatE aPHaRYngEatE CERCaRIa” (SCHIStOSOMatIdaE)
and tHE RECORdS Of ItS OCCURREnCE In BRazIL
aline Carvalho de Mattos; Bruno guimarães Lopes; Monica ammon fernandez; fábio fiebrig
Buchmann; Silvana Carvalho thiengo
Introduction: Schistosoma mansoni cercariae can be easily confused with other “Brevifurcate
apharyngeate cercaria” that also cause human dermatitis. Surveys of freshwater snails and
investigation on trematode-snail interactions in Rio de Janeiro State and areas of construction
of hydroelectric power facilities had been carried out by our team during the last years
aiming to know the snail vectors distribution in those areas as well as if they are infected by
S. mansoni or Fasciola hepatica. In this paper the occurrence of “Brevifurcate apharyngeate
cercaria” in six States is presented as well as its morphological discrimination from S. mansoni
cercariae. Methodology: In order to search and study the larvae, snails were isolated in jars
containing 4 ml filtered dechlorinated water. Live snails were exposed to an incandescent
light (60W) for six hours. After examination under stereomicroscope the samples were
maintained without light overnight to search for cercariae that emerge at night. Cercarial
stages recovered were observed under stereomicroscope, adding neutral red or lugol dyes
if necessary and subsequently identified. The infected snails were exposed to the light:dark
cycles as above weekly until a sufficient number of larvae were obtained for morphological
studies. Results: “Brevifurcate apharyngeate cercaria” was found in the following States:
Bahia (in Biomphalaria straminea and Plesiophysa guadeloupensis); Goiás (B. straminea,
Physa marmorata, Pomacea lineata, Drepanotrema lucidum and specimens of Ancylidae);
Mato Grosso (Pomacea insularum, P. lineata, D. lucidum and Biomphalaria amazonica; Minas
Gerais (Pomacea figulina); Pernambuco (B. straminea) and Tocantins (Potamolithus sp.,
Antillorbis nordestensis and B. straminea). As for the morphology “Brevifurcate apharyngeate
cercaria” presents furked tail and tail stem at least half as long as furcae, mouth at the
anterior end of body surrounded by oral sucker, intestine composed of two ceca, pharynx
absent, body without dorso-median finfold, uniform penetration glands, ventral sucker well
developed. The majority has eyespots and some of them may have furcal finfold. The S.
mansoni cercaria has no eyespots and furcal finfold. Conclusions: The morphological study
of “Brevifurcate apharyngeate cercaria” is important to discriminate S. mansoni cercaria
from those of other genus of this group as well as to help the surveillance and control of
schistosomiasis in endemic areas where both occurs. Furthermore, for effective control of
other associated zoonosis it is crucial that biological and taxonomic studies be continued
with the aim of matching the adult parasite with the larval forms via molecular techniques
(i.e. PCR, Sequencing) or those involving life cycles.
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MUnICíPIOS dO PaRanÁ OndE fORaM EnCOntRadaS aS tRÊS ESPÉCIES HOSPEdEIRaS
IntERMEdIÁRIaS dO SCHISTOSOMA MAnSOnI nO MESMO MUnICíPIO, nUM
UnIvERSO dE 210 PESQUISadOS
Jaqueline finau; antonio P. Carvalho; dourival Ribeiro da Silva
Secretaria Estadual de Saúde, PR - Brasil
Introduction: Schistosomiasis represents in Parana important public health problem,
particularly in the north of the state. A large number of environmental variables, population,
socio-economic conjunctures stimulated various migratory movements, crucial to the
spread of disease. The presence of molluscs, intermediate hosts, represents the potential
for transmission. Methodology: The localities were surveyed using GIS, with the purpose
of identification and characterization of the breeding, provided sketch bodies of water
sampled and their nature was recorded in proper form. During data collection, were used
shells perforated metal, with a long handle and the points determined at every 50 steps in
the container. The snails captured were placed in plastic bags, properly identified, containing
moist gauze. The method of capture followed the technical standards laid down in the
Manual of Surveillance and Control of Epidemiological Importance of Mollusks - Ministry of
Health, 2008. The technique for identification of the shellfish was performed according to
protocols established by Paraense (1961 and 1976) and Deslandes (1951) with adaptations
used by FIOCRUZ / René Rachou Research Center and has assembled a collection. Results:
We collected thousands of specimens of snails in the research stations, represented
by Biomphalaria occidentallis, B. peregrina, B. glabrata, B. intermediate, B. oligosa, B.
straminea and B. tenagophila, totaling 07 species of the genus Biomphalaria, and copies of
Drepanotrema, Physa and Lymnaea. In Assaí, Bandeirantes, Cornelio Procopio and Uraí, the
three species and intermediate hosts of Schistosoma mansoni were found in the same county
as well as having also been located in various regions of the state, including areas that are not
seen as risk to date. Conclusions: We have found the 03 known species as intermediate hosts
of schistosomiasis in Brazil, in the same county and in various regions of the state, raising the
need for precautionary measures and prophylactic measures.
266
natURaL InfECtIOn Of SCHIStOSOMIaSIS ManSOnI IntERMEdIatE HOStS In tHE
COaSt Of PERnaMBUCO
Mariana Izabel Sena Barreto de Melo; Constança Simões Barbosa; fábio Lopes de Melo;
Manuel amarista Sevilla
Centro de Pesquisa Aggeu Magalhães, Fiocruz, PE - Brasil
Introduction: Schistosomiasis mansoni is a parasitic infection caused by a trematode of
the genus Schistosoma. In Pernambuco, the endemic area historically known is Zona da
Mata whose Biomphalaria straminea is important snail species of S. mansoni’s transmition.
However, it has seen a disease expansion to state coast. By 2007 the mollusks species in this
area used to be B. glabrata. In 2008 a new malacological recognition identified B. straminea
in coastal town and in Recife’s metropolitan area. Methodology: Between September to
December 2008 was made malacological performed to diagnose Biomphalaria positiveness.
Snails was collected in coastal Pernambuco’s natural water collections, whose areas were
1 km maximum distance from the sea. The intermediate natural hosts infection rate was
determined by conventional diagnostic techniques of snails exposure to artificial light and
molecular techniques (PCR-nested). Results: Nine cities were selected and 97 points were
defined. We observed the presence of the intermediate schistosomiasis host in 45 collect
points, 11 of them inhabited by B. straminea, 34 by B. glabrata. In Paulista and Itamaracá city
were the coexistence of both species at four points (Janga and Forte Orange). B. straminea
was present in Itamaracá (Forte Orange), Igarassu (Mangue Seco), Paulista (Janga), Olinda
(Jardim Fragoso and Bultrins), Recife (Apipucos) and Cabo de Santo Agostinho (Gaibú and
Enseada dos Corais) cities. B. glabrata was found in Itamaracá (Forte Orange), Paulista
(Pau Amarelo and Janga) Jaboatão Guararapes (Dom Helder and Sotave), Olinda (Santa
Terezinha), Cabo de Santo Agostinho (Enseada dos Corais), Ipojuca (Porto de Galinhas) and
Tamandaré (Tamandaré). By the conventional method was identified natural infection rate
for B. straminea only in Paulista city (Janga). Among B. glabrata natural infection rates were
confirmed in Paulista (4.4% / Pau Amarelo and Janga), Jaboatão Guararapes (6.5% / Sotave
and Dom Helder) and Tamandaré (16.5 %). Through the molecular biology technique was
possible detect the DNA of Schistosoma mansoni in snails of 10 points in Ipojuca, Cabo de
Santo Agostinho, Jaboatão dos Guararapes, Recife, Paulista, Igarassu and Itamaracá. It was
obtained positivity results for B. straminea in 5 sapling points in Cabo de Santo Agostinho
(Enseada dos Corais), Recife (Apipucos), Paulista (Janga), Igarassu (Mangue Seco) e Itamaracá
(Forte Orange). Conclusions: The conclusion of the rates diagnosis infection in coast focus
allows us to understand the Schistosomiasis distribution in Pernambuco. Based in these
results is possible to plan an operation control by the local responsibles authorities.
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O PROgRaMa dE COntROLE da ESQUIStOSSOMOSE EM dOIS MUnICíPIOS da zOna
da Mata dE PERnaMBUCO: anaLISandO a IMPLantaçãO daS açÕES
Louisiana Regadas de Macedo Quinino; Isabella Chagas Samico; Constança Simões Barbosa
Introduction: The national schistosomiasis control has been implemented since 1975 by
the Superintendency of Public Health Campaigns (SPHC). In 1999, the decree No. 1399 has
normalized the decentralization of schistosomiasis control for states and municipalities.
Considering that the achievement of schistosomiasis control is influenced by the context
in which they are inserted and that, after decentralization no studies have been conducted
to evaluate how these actions are being undertaken by municipalities; this study aims to
know which factors, linked to political and structural-municipal context, can interfere with
the variation on the implementation degree of the Schistosomiasis Control Program (SCP)
in the municipalities of Escada and Goiana-PE. Methodology: For such, the normative
assessment on the dimensions, structure and process was carried out for determining the
implementation degree of control actions and, the evaluative research that has examined
the contextual elements, considering political and structural dimensions and their influence
on the implementation degree (ID) of the (SCP). It was built a logical SCP model from the
information contained in official documents. To collect data on the ID, it was designed a
structured questionnaire from the logical model applied to the SCP coordinators in the cities
studied. For collecting data on the context, another structured questionnaire was built from
the political and contingent model, which was applied to the health secretaries, coordinators
and agents of endemies from municipalities. Official documents were also evaluated and
direct observation was performed. It was used a scoring system that classified the ID of the
SCP in: deployed - when it reached 90 to 100 points; partially implemented - 60 to 89 points
and, not implemented - less than 59 points. Results: The SCP’s ID was not implemented
(52.85 points) in Escada and partially implemented in Goiana (63.65 points). The context
analysis showed that the main obstacles to the implementation of actions to schistosomiasis
control include few knowledge of players on the SCP operation and its indicators produced,
the poor planning and disintegrated of control actions, the low priority given to the
schistosomiasis and insufficient infrastructure. Conclusions: The findings point to a weakness
in the implementation of actions to schistosomiasis control in the municipalities.
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POLYPaRaSItISM BEtWEEn SCHISTOSOMA MAnSOnI and IntEStInaL PaRaSItES:
dEtERMInantS and gEOStatIStIC
Carla virginia vieira Rollemberg; fábio Jorge Ramalho amorim; Karla Carolline vieira
Rollemberg; Marília M. B. L. Silva; acácia M. B. Souza; Enaldo vieira; Evan W. Secor; guilherme
Loureiro Werneck; José antônio Pacheco de almeida; amélia Ribeiro de Jesus
Introduction: Mono and coinfection with schistosomiasis and intestinal parasites are
common in Asia, Africa and the Americas. Poliparasitism affects a substantial proportion
of the population and represents a major health problem because it, affects children’s
health and growth. However, few studies have been conducted addressing this issue.
The ability to predict the distribution of co-infections in large geographical scale has
important implications for the design of control programs for neglected tropical
diseases. Methodology: This study evaluates the epidemiology of Schistosomiasis
mansoni, helminths and protozoa infections, with particular emphasis on multiple
parasitic infections among 500 individuals who live in the riverside town and rice
producers of Ilha das Flores, Sergipe, Brazil. Individually georeferenced data from 500
subjects were collected by visiting 100 houses of four villages from the municipality
of Ilha das Flores (Bongo, Bolivar, Serrão and main city) from 2008 to 2010. The
questionnaires included clinical epidemiological and parasitological data (TF-test and
Kato-Katz methods). The geographical positioning of houses was determined using a
georeferenced cartographic database obtained from the Department of Planning,
Housing and Urban Development of Sergipe (SEPLAN). A database was built in the Spring
version 5.0.1. Parasitological samples distributions of the model coefficients were used
to produce maps for co-infection using Crimestat ® software by the estimator kernel to
produce risk maps. The prevalence maps were created in ArcView GIS Version 9 (ESRI,
Redlands, CA, USA). The analysis of association between S. mansoni infection and the
demographic, social, educational and environmental data were done using SPSS version
17.0. Results: The prevalence of the parasitic infections was: Trichuris trichiura (54.8%),
Ascaris lumbricoides (49.2%), Ancilostomideos (17.6%), Entamoeba histolytica (7.0%)
and Schistosoma mansoni (24.0%). Only 59/500 (11.8%) individuals did not present any
infection, whereas 279/500 (55.8%) had three or more parasites concurrently. A series
of co-occurrence of parasites, for example, 79/120 (65.8%) S. mansoni and T. trichiura;
61/120 (50.8%) S. mansoni and A. lumbricoides. Among the 120 patients with positive
stool examination for S. mansoni, 65.2% were male and 34.8% were female. Associations
between S. mansoni and being male gender (RP = 2.0, 95% CI 1.27 to 3.26, p = 0.003),
low education level (PR = 7.1, 95% CI 2.41 to 20.93, p <0.001) and low income (RP = 1.8,
95% CI 1.28 to 2.60, p = 0.0005). We also observed associations between S. mansoni
infection have any level of contact with natural water sources (PR = 1.9, 95% CI 1.36 to
2.59, p <0.001) and drinking untreated water (PR = 6.9, 95% CI 2.44 to 19.86, p <0.001),
with an association between degree III of water contact level (> 6 hours/week) and
infection as compared with degree 0 (no water contact); RP 3.6 IC95% [1.05 to 12.32]; p
= 0.04. Moreover, associations were observed between S. mansoni infection with being
a farmer (PR = 2.7, 95% CI, 1.29 to 5.40, p <0.001) or a fisherman (PR = 3.3, CI 95% 1.39
269
to 7.69, p <0.001). Conclusions: We conclude that the multiple parasitic infections in this
population is very common, which can also occur in other areas of Sergipe. The enriched
spatial statistical enables epidemiological factors associated and probable places of
contamination. The associations between S. mansoni infection with socioeconomic and
educational data call for an integrated approach to effectively control multiple parasitic
infections from the perspective of public health.
270
SCHISTOSOMA MAnSOnI PREvaLEnCE In StUdEntS aS an IndICatOR Of InfECtIOn
In tHE gEnERaL POPULatIOn In a HYPEREndEMIC RURaL aREa In MInaS gERaIS
StatE.
Kellen Rosa Coelho; Martin Johannes Enk; Leonardo ferreira Matoso; Mery natali Silva abreu;
Rodrigo Corrêa Oliveira; Helmut Kloos; andréa gazzinelli
Introduction: The current strategy for schistosomiasis control recommended by WHO is to
focus on high risk groups, such as schoolchildren. Studies with school-aged children as an
operational basis for the control of the disease showed that this group might be a good indicator
of Schistosoma mansoni infection in the whole population. However, although this strategy
has been identified as a possibility for effective control measure, it has not been recommended
in Brazil. This study aimed to evaluate schistosomiasis prevalence in grade school and high
school students as an indicator of infection in the total population in an endemic rural area in
Minas Gerais. Methodology: All households in São Pedro and Caju Districts in Jequitinhonha
Municipality and Virgem das Graças District in Ponto dos Volantes Municipality were
visited and invited to participate in the study. Demographic and socioeconomic data were
collected in all households using a questionnaire. Parasitological examinations were done
on two slides from three samples of stool each using the Kato-Katz method. Prevalence of S.
mansoni infection in individuals in each household was analyzed as the dependent variable
and prevalence of infection in students in each household, demographic and socioeconomic
parameters as the independent variables. Odds ratios and a chi square test, p≤0.05, were
used to evaluate associations between infection prevalence of individuals who do not live
with students and prevalence in the population. The Spearman, Mann-Whitney and KruskalWallis tests were used to evaluate the distribution of median prevalence among households.
The negative binomial regression was also used to explain how the independent variables
affect the dependent variable. Variables with p≤0.20 in the univariate analysis were included
in the final model. Results: Six hundred and seventeen students in the total population of
1,880 living in 469 households were included in the study. Prevalence of S. mansoni infection
in the total population was 55.1%, and higher among individuals 5-18 years (71.5%) and
among students (72.8%). Of the 469 households in the study, 209 (44.6%) had at least one
student who was positive, 202 (43%) had no students and 58 (12.4%) had students with
no infection. Of the 508 individuals living in households without students 198 (39%) were
infected. Prevalence of infection in these individuals was significantly correlated with overall
prevalence (OR=0.52; CI95% 0.42-0.64; p<0.001). Prevalence of infection in individuals who
lived with an infected student was 68.8% and for those who lived with a negative student
25.2% (p<0.001). Presence of at least one positive student in the household increased the
likelihood of infection 2.22 times (CI95% 1.11-4.43). In the household analysis prevalence of
infection among students was significantly correlated with the prevalence of infection in all
individuals in each household (Spearman coefficient 0.70; p<0.001). Level of education of
households heads, number of appliances and household income were inversely related to
total household prevalence. The comparisons of the medians of total household prevalence
with the socioeconomic variables occupation of household head, ownership of a car and/or
motorcycle, presence of faucet, tank, shower, bathroom and water container were strongly
271
associated. In the univariate analysis, the students’ prevalence of infection per household
obtained a coefficient of 0.01 (CI95% 0.009-0.012), meaning that for every 1% increase in
the students’ prevalence of infection, the expected probability of overall prevalence per
household increased by 1%. The largest negative coefficient was found with the variable
ownership of car and/or motorcycle (coef. -0.49; p=0.002), indicating that overall prevalence
per household decreased by 49% in families that owned motor vehicles compared to
families lacking them. Whereas the number of persons per room in the house was positively
correlated with prevalence of infection (0.17, p=0.020), the number of appliances and the
presence of faucet, tank and water container was negatively correlated. In the final model
the variables number of appliances (coef. -0.05), motorized vehicle ownership (coef. -0.34)
and student’ prevalence (coef.0.01) remained significantly correlated with the prevalence of
infection in all individuals in each household. Conclusions: The results show that students
can be an acceptable indicator of S. mansoni infection in households in São Pedro, Caju and
Virgem das Graças Districts, suggesting a higher chance of finding positive individuals in
households with positive students. The prevalence of infection in students was associated
with socioeconomic factors, an important predictor of overall prevalence at the household
level. This study may help guide new, alternative strategies in schistosomiasis control in
areas with limited resources. Financial support: CNPq, NIH-ICIDR Grant 1R03AI071057-01,
FAPEMIG, CAPES.
272
SCHIStOSOMIaSIS and ECOSYStEM aPPROaCH tO HEaLtH: a SYStEMatIC REvIEW.
Marisa da Silveira Soares; Margareth Maria Lessa gonçalves; Magali gonçalves Muniz Barreto
Iinstituto Oswaldo Cruz, Fiocruz, RJ - Brasil
Introduction: The concept of “Ecosystem Approach to Health” (EAH) reports to genuine
participative processes of management that anticipate changes and create adaptive
solutions for complex systems of the interface health/environment, which ask for nonlinear treatment. To approach health in such way means to search for the sustainability
for communities as for ecosystems, in contexts with ecological, social and health problems
interacting in different scales (familiar, local, municipal etc.), which demands processes
of negotiation between the different “stakeholders” involved and, thus, a participative
and democratic attitude of the scientists. In the scope of shistosomiasis, this approach
can represent the overcoming the biomedical paradigm strong limitation to deal with
the surveillance and the control in facing the extreme complexity that can take place
in the epidemiological situation. So, the identification and the analysis of previous
experiences based on methodologies similar to an EAH (systemic, participative, focused
on ecosocial sustainability etc.), according to the theoretical-methodological debates on
this approach, can contribute for the development of new experiences, more robust in
the understanding and in the management of complexity. The objective of this work is
to verify if the contents of the papers obtained in a systematic review on approaches of
schistosomiasis (integrated, participative, ecosocial etc.) are close or apart to the concept
of EAH. Methodology: The review was based on the bibliographical databases of the
Virtual Health Library (BVS) (Scielo, MEDLINE, Cochrane Library etc.), on the Information
Sciences Institute (ISI) and on the International Symposium on Schistosomiasis Abstract
Books. The searches in BVS and ISI were carried out with the key-word “schistosomiasis”
combined with the expressions “ecosystem approach”, “integrated approach”, “ecosocial
approach”, “socioecological approach”, “ecological approach”, “participative approach”,
“popular participation” and “popular education”. The approximation in relation to the
EAH was investigated with the aid of questions such as: is it really a complex situation? Is
the approach systemic? Does the process demonstrate concerns with the sustainability of
ecosystems and communities? Does the work consider different scales? Does the process
anticipate changes? Does the process create adaptive solutions in a participative way? Does
the work recommend the participation of science in the negotiations between interests by
presenting future scenaria narratives to guide the decision process? Results: None of the
184 articles or thesis found perfectly fits the criteria of compatibility with the EAH concept.
On the other hand, many works - mainly in anthropology and primary attention to health
- although far from the EAH according to certain criteria, are close to it according to other
important ones, such as participative approach, a new posture for scientists and a new
role for science. Others present broad methods, nevertheless cannot be considered as
systemic. At last, the research found works that have no relation at all with an EAH, since
they are strict to a single discipline, dealing with vaccines, moluscicides, diagnosis and
other subjects, frequently with experimental approach. Concerning abstracts, even though
only one, published in 2005, was specifically about EAH, many were close to the concept
according to essential criteria. Conclusions: It is still necessary to improve the process of
273
construction of this field of the scientific knowledge in schistosomiasis scope. For this,
investments are needed in the familiarization of scientists with the EAH concept, mainly
for the appropriation of this subject by the segment of the scientific community that guides
public policy and decision processes in the interface health-environment. This investment
is challenging, since, to opt for this approach, scientists have to abandon some of their
historical prerogatives and to accept the contribution of other disciplines and knowledges.
274
SCHIStOSOMIaSIS In PatIEntS attEndEd at tHE HOSPItaL Of tHE fEdERaL
UnIvERSItY Of aLagOaS In MaCEIO (aLagOaS- BRazIL)
thiago andré alves fidelis; thiago de Oliveira assis; Janira Lúcia assumpção Couto; Rozangela
Maria de almeida fernandes Wyszomirska
Introduction: Infections caused by trematode Schistosoma mansoni, reaches high levels in
the state of Alagoas, in where it causes serious problems in public health. Data from the
Schistosomiasis Control Program in Alagoas (2007) refers that 69% of the territory is endemic
and about one million people would be exposed to infection. Alagoas has the highest number
of positive parasitological examinations among the states in the Northeast and Southeast
regions. In this context, the objective of this study is to evaluate the profile of patients with
schistosomiasis reported from Academic Hospital (Federal University of Alagoas), in Maceio
(Alagoas-Brazil). Methodology: This is a retrospective study, collecting data from medical
records of patients diagnosed with schistosomiasis, in a period of five years (2006-2010). The
data were stored in a software Bioestat v. ® 5.0 database’s and was applied simple descriptive
statistics to quantitatively analysis. Results: Analysis was performed on medical records of
108 patients with a positive diagnosis for schistosomiasis, 9 were admitted to the hospital
services in 2006, 42 cases of hospitalization in 2007, 39 cases in 2008, 11 in 2009 and 5 in
2010. Considering 108 patients, there was discrete predominance of males, representing
57.4%. The youngest found was 18 years old and the oldest 84, and the average age was
55.8 ± 15.6 years. Most cases came from Maceió (56.4%), while 7.4% came from Rio Largo
and 3.7% from União dos Palmares. The clinic form that was most found was hepaticsplenic
(92.5%) and hepaticintestinal (6.4%). Conclusions: The results showed heterogeneity in
affected age due to a high standard deviation. There are a greater number of patients from
Maceió, but there was insufficient data to clarify that they came from other cities. The
majority of patients have hepaticsplenic clinical form, observed in a sample of patients from
hospital.
275
SCHIStOSOMIaSIS In tHE tOURISt CItY Of HOLaMBRa (SãO PaULO, BRazIL), fROM
1993 tO 2008
Marisa da Silveira Soares; Celia Maria thomé; Cesar Luiz Pinto ayres Coelho da Silva; Claudia
Portes Santos Silva; Magali gonçalves Muniz Barreto; denise de assuncão Borges; Rita Maria
Silva; Cybele gargioni; vera L. M. Oliveira; Herminia Y. Kanamura
Introduction: The touristic city of Holambra, located in the Metropolitan Region of Campinas,
SP, Brazil, presents autochthonous cases of schistosomiasis, immigrants coming from endemic
areas, has breeding areas of Biomphalaria tenagophila and poor sanitation. The objective of
this work was to investigate the epidemiological situation of schistosomiasis in Holambra,
with emphasis in the search of knowledge for predicting future scenaria. Methodology:
All epidemiological data collected from the institutions working with shistosomiasis in
Holambra since the foundation of the city (Municipal City Hall, SUCEN, IALutz, FIOCRUZ
etc.) were analyzed. For the study on morbidity, we used SINAN’s database and other
documents filled since the foundation of the city in 1993 until 2008. All data were achieved
by spontaneous demand, periodic examinations, coprologic and seroepidemiologic censuses
and active search, carried through quarters, companies, schools and institutions of health.
The stool examinations (Lutz, Kato-Katz and Ritchie) followed varied arrangements, with
up to three samples and 12 microscope slides per person. Serology was based on indirect
imunofluorescence for research of IgM antibodies (IFT-IgM), with confirmation of the positivity
by stool examinations. Each case was identified according to the sex, age, occupation, place
of birth and autochthony. For the censuses, prevalences were calculated, but the scarcity
of clinical data hindered analyses on intensity and forms of infection. Associations of the
epidemiological data of censuses were analyzed by statistical test (qui-square or Fisher;
program Epi Info 6,0). The hydric collections were investigated according to the presence
of sewers in natura, attendance of population and presence of snail intermediate hosts of
Schistosoma mansoni. The data on snails corresponded to the sporadic or regular collections
in water bodies with epidemiological importance. The tests of natural infection in snails were
carried out by crushing and exposition to artificial light. Results: In the whole period of the
study, there were 171 human cases of infection by S. mansoni, of which 157 inhabited in
Holambra and 14 only worked there; 115 (67.3%) were males and 56 (32.7%) were females
and, 150 (87.7%) were above school age. The 110 agricultural workers represented 65.9% of
the 167 cases with information on occupation and when considered the information on the
classification, 87 (52.1%) were autochthonous, 25 (15.0%) were imported and 55 (33.0%)
were indeterminate. Considering only the last five years there were 27 autochthonous cases,
while seven were imported and 18 were indeterminate ones. Most of the cases were from
the states of São Paulo and Paraná. There were 34 cases detected by occupational medicine
service, confirmong the occurrence of occupational transmission. In regard of the results of
each census, the coprologic one carried through the years 2000 and 2001 showed 6,9% of
positivity (N= 725), affecting mainly males (pχ² = 0,0124) and individuals above the school
age (pχ² = 0,0000). Of the 50 cases, 18 were inhabitants and workers in a company that
did not adopt periodic examinations. In another census performed in this exactly quarter in
2003, the prevalence was of 2,3%, with more cases in the same age group (pχ² = -, 0142). The
276
seroepidemiological census in a school showed nine positive cases by serology (prevalence
of 20,0%), two of them were confirmed by coprology (prevalence of 4,4%). In another school,
the prevalence by serology was 13,3% (11 cases), three of which were confirmed by coprology
(prevalence of 3,6%). Concerning the investigation of local conditions for transmission, 13 of
the 71 places of human contact with natural waters were breeding places for Biomphalaria
tenagophila, one was for B. occidentalis and there were both species in another. All of them
received sewers. Snails positive for S. mansoni infection were found only in 2001, in the
place more visited by the patients for leisure, the same that in 2007 presented simpatry
of B. tenagophila and B. occidentalis. Other trematode cercariae, including a potential
antagonist to Schistosoma mansoni, were found infecting the snails Conclusions: The results
show that Holambra is an endemic area of low prevalence of schistosomiasis tending to have
outbreaks of cases, which is conflicting with the status of “Touristic City”. The complexity of
its epidemiological situation requires suitable mechanisms for monitoring the peculiarities
of low endemicity and to detect schistosomiasis cases in groups that are rarely considered
for public policy purposes (people above school age, rural workers not resident in the city,
migrant workers). The Family Health Program and the sectors of the economy related to
these workers have an important role to play in the local surveillance of schistosomiasis.
277
SCHIStOSOMIaSIS ManSOnI In CHILdREn and YOUtH Of a MUnICIPaL dIStRICt Of
aLagOaS (BRazIL)
Janira Lúcia assumpção Couto; Michelle Martins da Silva; nathalie Louise Santos de Oliveira;
Elaine Cristina e Silva; Mykaella andrade de araujo; Jefferson alain da Costa ferreira; thiago
andre alves fidelis; alinne Schristina da Silva; Pedro filippe Simões Santos; francisnaldo
Emanuel nunes P. torres
Introduction: The high levels of Schistosomiasis mansoni in Northeast region of Brazil result
mainly from ambient conditions where the population lives. Alagoas state has 102 cities and
70 of them are endemic areas. Although data of the Health State Secretary (2007) affirms
a reduction in the prevalence after the action of the control program, it verifies that not
yet it has adequate housings, adjusted sanitation, nor adequate destination of the garbage
and dejections. In this context of disequilibrium between population and environment, the
schistosomiasis spreads indefinitely. This work searched the schistosomiasis occurrence in
children and youth in Marechal Deodoro city, located in southeastern region of Alagoas
state. Methodology: The research happened in the period from March to July 2010, with the
group of study formed by children and youth with age between three and 14 years. Initially,
it was evaluated relative aspects about how they live, by means of inquiries with questions to
the housing. The parasitological analysis were effected by the methods of Lutz and Kato-Katz.
The nutrition state was evaluated through measures of the weight, height and age. Results: It
was examined 178 children and youth, 65 of them (36.51%) had Schistosoma mansoni e 113
(63.48%) had diverse intestinal parasites, between protozoa and helminthes. The parasitic
load of S. mansoni was above of 100 eggs/g of faeces in 35 individuals (49.23%), above of
500 eggs in seven (10.76%) and above of 1000 eggs in three (4.61%). The houses, where
they live, were made of bricks (95.38%), and had sewers system (64.61%). The nutritional
evaluation showed some cases of obesity (15.38%) and malnutrition (7.69%). The treatment
of schistosomiasis and other parasites was provided with medical prescriptions and
medicines of the local Health Secretary. Conclusions: The high levels of schistosomiasis and
the high parasitic load verified can indicates a hiperendemic area. It seems the houses are
adjusted, however it exists sewers in open air and flooded streets, what it turns unhealthy
the environment. The common habit of walking bare-footed facilitates the infection. These
data found in the city become necessary urgent measures for the control of the endemic
disease, mainly a adjusted basic sanitation.
278
SnaIL SPaCIaL dIStRIBUtIOn In UBERLandIa - Mg‘S URBan zOnE
amaral alves de Souza; alessandro ambrosio dos Reis; Lara Lane de Oliveira; adalberto
albuquerque Pajuaba neto; Elisângela de azevedo Silva Rodrigues; Márcia Beatriz Cardoso
de Paula
Prefeitura Municipal de Uberlandia, MG - Brasil
Introduction: The schistosomiasis, one of the most widespread human parasitosis in
the world, generally occurs in the basic sanitation absence or precariousness. Currently,
it shows a clear tendency to expand due to the human migrations from endemic areas.
The snails occurence record of Biomphalaria genus is very important because this group
aggregate species of snails‘ which act as intermediate hosts of Schistossoma mansoni.
Methodology: using GPS service, 14 streams of Uberlandia - MG‘s urban zone were mapped
and punctuated. Them those streams were surveyed regarding snails presence. When
found, they were collected using shells, properly packed in plastic container and sent to
the zoonosis control center lab for examinations using induction methods. Results: during
the research period 265 specimen of snails were captured in nine streams. Them, 147 were
identified as Biomphalaria straminia, 110 were identified as Biomphalaria tenagophila and
eight were identified as Biomphalaria glabrata, Schistossoma mansoni potencial tranmitter.
All the examined specimen were negative to schistose. Conclusions: despite the fact that
Uberlândia has a great infrastructure and basic sanitation is necessary the supervision
actions maintenance in the places where the Schistossoma mansoni has the possibility of
completing its life cycle due to the human migrations from endemic areas.
279
SPatIaL anaLYSIS Of SCHIStOSOMIaSIS HUMan CaSES at Santa MaRIa
nEIgHBORHOOd, aRaCaJU-SERgIPE.
Karina Conceição gomes Machado de araújo; Helder Barreto Silva Junior; Roseli La Corte dos
Santos; andréa valença Cardoso
Introduction: Schistosomiasis is a endemic disease in several cities of Sergipe. According
to data from Health State Secretary about the schistosomiasis contention actions, they are
integrated to the activities of the Family Health Program (Programa Saúde da Família). In the
last years, the prevalence rates are keeping stable, without relevant changes in the diseases´
transmission indexes. Human health and environment health are deeply associated and
this could be observed in the transmission dynamics of hidric vehiculation diseases, such
as schistosomiasis. In unprovided sanitation infrastructure areas, population usually make
use of the streams and other natural hidrical collections to several purposes, like disposal of
sewage, domestic use, agropecuarial activities and leisure. This study aimed to analyze the
spacial distribution of human cases of schistosomiasis in Santa Maria neighborhood in the
city of Aracaju-SE. Methodology: The study area was the Santa Maria neighborhood, located
in the southern of Aracaju. Descriptive data were acquired at the Laboratory of Parasitology,
Department of Morphology of Federal University of Sergipe, where research has been
conducted on the distribution of prevalence for schistosomiasis in that area. Absolute method
was used with instantaneous positioning of a point, with the aid of a GPS receiver for the
spatial location of the participants´ houses. The availability of data was performed through
a query system based on Geographic Information System (GIS), which allowed storing all
the spatial data manipulation and linking of descriptive attributes to the graphic features,
allowing the visualization and spatial analysis of data in the system. Results: Patients who
had some type of parasite were referred to the health family unit for proper treatment.
Cases of S. mansoni were sent to the Health Secretary of the city, where those responsible
were responsible for making the notification and treatment of disease. The database of
Santa Maria consisted in graphics and descriptive data and these were inserted into the table
of TERRAVIEW. Were mapped, in the location studied, 272 cases of schistosomiasis, but only
225 were treated. Descriptive data were represented by cases of schistosomiasis found in
the period 2009 to 2010. The thematic map generated showed a uniform spatial distribution
of disease cases in the locality studied. Conclusions: The community of Santa Maria district
is naturally a place lacking in basic programs in health education, which may be generating
the persistence of cases of schistosomiasis. The data from the present study do not allow
estimating the emergence of new cases, but tracking them through the GIS, annually, keeping
always updated information for epidemiological surveillance of schistosomiasis.
280
SPatIaL anaLYSIS Of SCHIStOSOMIaSIS
nEIgHBORHOOd, aRaCaJU-SE.
vECtOR
fOCI
In
Santa
MaRIa
tiago Pinheiro vaz de Carvalho; Roseli La Corte dos Santos; Constança Simões Barbosa; Karina
Conceição gomes Machado de araújo
Introduction: The maintenance of Biomphalaria foci, the intermediate host of Schistossoma
mansoni, is related to various environmental conditions with emphasis on rainfall annual
changes. Thus, during the dry season when water flow decreases and causes the emergence
of pools, there is a higher concentration of snails, increased human access and contact with
infected water. In rainy season the volume and speed of flow of waters carry the snails to
other habitats. Whereas most of the information needs of managers in local governments is
linked to a geographic location, the dynamic use of maps is a useful process to make more
effective the taking of decisions in public health with great impact on the Unified Health
System (UHS). This study aimed to analyze spatially the distribution of schistosomiasis foci in
Santa Maria neighborhood in the city of Aracaju-SE. Methodology: The malacological raising
was accomplished through systematic collection, by sweeping, of schistosomiasis´ snails in all
natural or artificial water collections found. Descriptive data for performing spatial analysis
(street, spatial location of foci, number of snails examined and positive, rate of infection
for S. mansoni) were acquired through field research. Absolute method was used with
instantaneous positioning of a point, with the aid of a GPS receiver to the spatial location
of the breeding of the intermediate hosts of Schistosoma mansoni and potential sources
of transmission. The database was constructed using the Epi-Date and analyzed in Epi-Info.
The maps were constructed and analyzed in the software TERRAVIEW. Results: The group
of investigations performed highlighted the importance of the areas surrounding homes as
a risk factor in the transmission of schistosomiasis in the Santa Maria neighborhood, since
most of the foci were located peri or intra-domiciliary. The finding of Biomphalaria glabrata
in the locality studied, it is common and results from inefficient urbanization in these areas,
where they are located ponds, streams, ditches, puddles and ponds harboring populations
of this mollusk. The presence of the snail host in some hidrical collections, in areas not fully
urbanized of the studied locality, reflects a concern about the possible occurrence of new foci
of schistosomiasis. However, data from this study do not allow estimating the emergence of
new foci, but tracking them through the Geographic Information System, annually, keeping
always updated information for epidemiological surveillance of schistosomiasis. Conclusions:
The identification and spatial/temporal location of areas where there are different degrees
of risk for transmission of schistosomiasis in the Santa Maria neighborhood, can help to
guide decisions for planning health interventions.
281
SPatIaL dIStRIBUtIOn Of dEatHS fROM SCHIStOSOMIaSIS In PERnaMBUCO, fROM
2005 tO 2008
Bárbara Morgana da Silva; Silvânia aa Lima; Eduardo n albuquerque; neusa E Magalhães;
nara Pedrosa arruda
Secretaria de Saúde de Pernambuco, PE - Brasil
Introduction: Schistosomiasis mansoni is a disease caused by the trematode parasite
Schistosoma mansoni presenting initially be asymptomatic and may progress to chronic
and sometimes causing death. In Brazil, reported about 500 deaths a year, Pernambuco
responsible for 37% of them. The use of the spatial distribution of events in health can best
express the different access to goods and services and health infrastructure, the location of
potential environmental risk and the areas where concentrated vulnerable social situations,
replacing a model of individual risk. To describe the spatial distribution of deaths due to
schistosomiasis in the State during the period 2005 to 2008. Methodology: We conducted an
ecological study, descriptive, using data from the Mortality Information System, Information
System for Notifiable Diseases, DATASUS and SIS-PCE. Results: Of the 185 municipalities of
Pernambuco, 126 (68.11%) recorded deaths from schistosomiasis during the period 2005 to
2008, totaling 780 cases. Among eleven Regional Offices only the seventh and ninth reported
no cases. The deaths are concentrated in the coastal / forest area of the state, considered
endemic, where the first, second and third Regional with 51.06%, 19.74% and 14.61% of
deaths respectively. Whereas deaths from infectious and parasitic diseases, schistosomiasis
represents the state, the rate of 7.75%, highlighting the GERES II and III with 16.00% and
15.36%. Conclusions: As expected the concentration of deaths was located in the epidemic
area of the state, alerting managers to the quality of the Schistosomiasis Control Program.
In addition to the actions of health surveillance and patient care, it is necessary interagency coordination in the areas of infrastructure and education that positively modify
the conditions that favor transmission and maintainers of the localities where ecological
conditions are favorable and space organizations.
282
SPatIaL dIStRIBUtIOn Of SCHISTOSOMA MAnSOnI InfECtIOn In adOLESCEntS
UndERgOIng CHEMOtHERaPY WItH PRazIQUantEL In an EndEMIC aREa Of
SCHIStOSOMIaSIS In PERnaMBUCO, BRazIL
aline favre galvão; Oswaldo gonçalves Cruz; tereza Cristina favre; Otávio Sarmento Pieri
Introduction: Schistosomiasis is one of the most prevalent parasitic diseases worldwide,
and praziquantel (PZQ) chemotherapy the main strategy adopted for its control. Because
this endemic disease is determined in space and time by environmental risk factors, spatial
analysis and GIS are important tools for better understanding its transmission and distribution.
In this context, there is a lack of information about changes in spatial distribution pattern of
prevalence of infection after drug administration, which can contribute to improve actions for
its control. Methodology: A double-blind randomized clinical trial was conducted to evaluate
the impact of PZQ 60 mg/kg single dose treatment on S. mansoni infection compared to
the PZQ 40 mg/kg standard dose in adolescents from São Lourenço da Mata. A preliminary
parasitological survey selected individuals with more than 100 eggs per gram of faeces by
the Kato-Katz method. Those suited to the inclusion / exclusion criteria were recruited and
treated with 40 mg/kg or 60 mg/kg PZQ single doses. Monitoring parasitological surveys were
performed at 21, 180 and 360 days post-treatment. Prevalence and intensity of infection
were compared throughout the study in both treatment groups using contingency tables (chisquare or Fisher‘s exact test) and ANOVA. The influence of other variables in the study was
assessed by logistic regression analysis. A malacological survey was conducted to determine
the natural infection of the local intermediate host specie, Biomphalaria straminea. The
spatial distribution analysis of infection in both groups before and after (180 and 360 days)
treatment was performed by kernel density estimates for case cluster detection. Results:
Altogether, 123 adolescents (10-19 years) were examined in four study stages. Although all
patients were negative 21 days after treatment, 17.9% and 30.9% were positive at 180 and
360 days, respectively. The monthly natural infection rates of B. straminea ranged from 0 to
2.2%. Kernel analysis results confirmed the parasitological data, allowing the mapping of risk
areas based on “hot spots” identified. Conclusions: As the fecal contamination of freshwater
aquatic environments is the main component of the schistosomiasis transmission process, it
is recommended that chemotherapy be combined with preventive control measures, such
as water supply and environmental sanitation to reduce human contact with intermediate
host breeding. Control of Biomphalaria populations through local small-scale measures,
such as drainage, vegetation and debris removal from ditches and streams to become
unsuitable for breeding or maintenance of clams populations, and information, education
and communication strategies should also be encouraged and given priority, aiming the
sustainable control of disease.
283
SPatIaL-tEMPORaL anaLYSIS On fOCI Of SCHIStOSOMIaSIS tRanSMISSIOn In PORtO
dE gaLInHaS, PERnaMBUCO: a PROCESS Of EndEMIzatIOn ?
Elainne Christine de Souza gomes; Onicio Batista Leal neto; Karlla Priscila Monteiro viana;
Isabela Regina alvares da Silva Lira; Rafael Cesar Lima Pedroso de andrade; diego Leandro
Reis da Silva fernandes; Julyana viégas Campos; Hilda Carla Moura dos San; Constança
Simões Barbosa
Introduction: Schistosomiasis is a health hazard that in Pernambuco is considered to be
characteristic of rural areas, but it has been expanding to coastal localities such as Porto de
Galinhas. Since the year 2000, foci of the vector molluscs that produced acute cases of the
disease have been recorded in this region. With a view to understanding this epidemiological
process better, a wide-ranging epidemiological survey is being conducted at this locality, 10
years after the abovementioned events. The present study had the aims of (1) carrying out a
georeferenced malacological survey; and (2) constructing thematic maps in order to compare
the distribution of the current foci of Schistosoma mansoni with those recorded 10 years
earlier at the locality of Porto de Galinhas. Methodology: (1) The malacological survey began
in July 2010 with mapping of the locality using a GPS (Global Positioning System). Homes,
streets and water bodies with the presence of the mollusc vector were georeferenced on
the template. Seven localities that form part of Porto de Galinhas were mapped (Vila de
Porto, Merepe I, II and III, Salinas, Socó and Pantanal), and an active search for breeding sites
of Biomphalaria was conducted at these localities. Snails were gathered over a 15-minute
period using scoops and tweezers, and were stored in plastic tubs for transportation to
the Schistosomiasis Laboratory of the Aggeu Magalhães Research Center, Fiocruz. From
each batch, 10% of the specimens were removed randomly for dissection and taxonomic
identification of the species. To diagnose S. mansoni infection and thus identify the foci of
transmission, the snails were exposed to artificial illumination to induce emission of cercariae.
The snails that were found to be negative using this technique were then crushed to search
for sporocysts. (2) Thematic maps representing the breeding sites and foci of B. glabrata
were constructed using the GPS TrackMaker software. The current spatial presentation of
the breeding sites and foci were plotted on these maps. Results: During the first collection
of the malacological survey, 3.747 snails were caught and 35 breeding sites of the species
B. glabrata and five foci of schistosomiasis transmission were identified. Fig. 1 shows the
preliminary results from this survey, with the geospatial distribution of the breeding sites
and foci among the different localities of Porto de Galinhas: Merepe III (1.077 snails were
collected from five breeding sites and 1 focus), Salinas (2.466 snails from 24 sites and 4 foci),
Socó and Pantanal (204 snails from 1 site) and Merepe I, II and Vila de Porto de Galinhas (no
breeding sites identified). Comparison between the records from this first collection (July
2010) and the collections made in August 2000 (Fig 2) showed that the number of foci at
Merepe III had gone down (from nine to one), while Socó and Pantanal presented two and
one foci, respectively. Only the locality of Salinas presented an increase in the number of
foci (from three to four). The high number of breeding sites identified at this locality, in
association with the precarious conditions of basic sanitation, housing and hygiene, indicate
that there is the potential for new outbreaks of the disease, given that under appropriate
284
environmental conditions, breeding sites for B. glabrata are rapidly transformed into foci
of schistosomiasis transmission. If heavy seasonal rains occur, the infected snails will be
carried into the streets and backyards, thereby exposing the entire population of Porto de
Galinhas. Further collections of molluscs will be undertaken in order to identify other foci.
The analytical resources of the Geographic Information System (GIS) will make it possible to
generate thematic maps with greater refinement of the epidemiological attributes, such as
gradation of the vector mollusc infection rates for each georeferenced focus. Conclusions:
The reduction in the number of foci in the Merepe III district was associated with urban
improvements, coming from asphalting many streets, thereby eliminating the breeding sites.
The heavy rainfall that occurred in June and July 2010 explain why no breeding sites and foci
were recorded in other localities, since the snails were taken away by the torrents of water.
Further collections and investigations will be undertaken during the period subsequent to the
rainy season, a time at which the breeding sites will be more stable, which will favor mollusc
reproduction and increase the chances of mollusc infection. The foci identified now, added
to those recorded in 2000, confirm that disease transmission has been maintained over the
last ten years and indicate that this health hazard has become endemic in yet another coastal
region of the state of Pernambuco. Financial support: CNPq
285
tHE CHangIng RISK In tHE EPIdEMIOLOgIC PROfILE Of SCHIStOSOMIaSIS In
ROndÔnIa StatE
flávia Serrano Batista; Carlos antônio amante; Sônia Maria dias de Lima; Maria de nazaré
Reis alves
Secretaria Estadual de Saúde, RO - Brasil
Introduction: The State of Rondônia is formed by 52 towns, and among them, at least 15
have a high incidence of land and aquatic snails, among them we can mention the huge
African snail Achantina fulica, some species non-identified of Biomphalarias, B. peregrina
and B. ocidentalis. The relation of points of the malacological research is little due to a
reduced number of trained malacologists and the epidemiologic importance that the
disease still has in the State. Among all the notifications that were carried out in the State
of Rondônia, none of them was identified as an autochthonous case, and it happens due to
the fact of many individuals who live here had immigrated from endemic regions, and these
are considered alochthonous cases. The increasing of population has happened very fast and
disorganized in the State of Rondônia. In 1998 the total population was 1.276.181, and in
2008 it was estimated in 1.493.566. Because of the process of installation and construction
of the hydroelectric plants in Porto Velho there was a disorganized increasing of about 60%
more than the current population. So, there was an increasing of flooded regions in the
city and, because of the arrival of immigrants from endemic regions, it is believed that an
epidemiologic changing has been happening in the State. Ouro Preto do Oeste is a town in the
State of Rondônia located at BR 364 330 km from Porto Velho. It has a territorial area of 4.975
Km ² and an estimated population about 41.000 inhabitants. It was carried out a malacologic
work in 100 points during 20 years and these data helped the program of schistosomiasis
controlling, not only for the malacologist’s experience, but also for the state. This organized
relation of points of malacologic research includes regions like small rivers, smallholdings,
farms, neighborhoods, plots, weirs, small lakes with the presence of existing snails, and in
this region the number of schistosomiasis cases is much bigger than in other towns of the
State. Methodology: This paper is entirely retrospective, and it was used stored data in the
program of schistosomiasis controlling of the State of Rondônia in Agência de Vigilância em
Saúde – AGEVISA (Surveillance Agency in Health) which were kindly lent by Carlos Antônio
Amante (FUNASA), a malacologist who had helped to store these data. Results: Among 100
researched points in this region, in 28 of them were found Biomphalaria amazonica and
Biomphalaria ocidentalis, among other ones. The Laboratory of Medical Helminthology and
Malacology, René Rachou Reseaching Center from Belo Horizonte had been recently in two
towns of the State and carried out malacologic collection in order to confirm the suspect
of Biomphalaria cousini which was the same one in the previous described morphology
in the State, Biomphalaria amazonica. This is a worrying fact when the previous results of
liability to Schistosoma mansoni show that B. cousini becomes infected with this parasite,
according to the Laboratory of Medical Helminthology and Malacology. Even without the
results of this recent collection, it is important to point out the arousing for potential ways of
schistosomiasis transmission. Results: Among 100 researched points in this region, in 28 of
them were found Biomphalaria amazonica and Biomphalaria ocidentalis, among other ones.
The Laboratory of Medical Helminthology and Malacology, René Rachou Reseaching Center
286
from Belo Horizonte had been recently in two towns of the State and carried out malacologic
collection in order to confirm the suspect of Biomphalaria cousini which was the same one in
the previous described morphology in the State, Biomphalaria amazonica. This is a worrying
fact when the previous results of liability to Schistosoma mansoni show that B. cousini
becomes infected with this parasite, according to the Laboratory of Medical Helminthology
and Malacology. Even without the results of this recent collection, it is important to point
out the arousing for potential ways of schistosomiasis transmission. Conclusions: In this way,
it is necessary to arouse a lot of important points, among them the necessity of increasing
specific trainings for malacologic research and the survey of planorbydic letter in the state
of Rondônia because the potential host presence data in the schistosomiasis transmission is
still small and we need this research to accept the epidemiologic changing that has occurred
in the last years, mainly because of the arrival of immigrants from endemic regions, and so,
set up surveillance strategies to control the cases of the disease in the state. The support
of the research laboratory is essential for this happens, since there aren’t equipments of
high complexity to identify the snails. Resuming the schistosomiasis controlling program is
indispensable in this moment of fast increasing for public health conversation in the state.
287
tHE COMPLExItY Of SCHIStOSOMIaSIS In HOLaMBRa (SãO PaULO, BRazIL): CHECKIng
BaSES fOR an ECOSYStEM aPPROaCH tO HEaLtH
Marisa da Silveira Soares; Marcela C. Mansano; Magali g. M. Barreto; Marcelo f.S. Porto
Introduction: Holambra is a Tourist city of one of the richest Metropolitan Regions of
Brazil (Campinas, SP), where schistosomiasis is endemic. Its economy, based mainly on the
floricultura, has contributed for the circulation of migrantes workers and families at the state
of São Paulo, many of which deriving of endemic areas of this parasitosis. This conjunction of
factors makes the epidemiological situation of schistosomiasis in Holambra important for the
local, state and national collective health. The expectation of the Unified Health System (SUS)
in the city in 2008 was to get the state certification for the elimination of schistosomiasis. This
goal requires dealing with an “emergent complexity”, produced mainly by peculiarities of the
low endemicity, by the complexity of the intricate intermunicipal and interstate migratory
processes of agricultural workers and families in that Brazilian state, and also by uncertainties
and conflicts of interests expected in production processes that occur in a context in which
agribusiness depends on unhealthy water sources. The Ecosystem Approaches to Health look
after this requirement, because deal with the complexity of the interface health-environment
in its aspects not reductible to mathematical-computational models, such as ethical and
political aspects, allowing a consistent decision process. But, to adopt them, it is necessary
to assume the essentiality of seeking sustainability of ecosystems and communities, dialogue
between disciplines and sectors, genuine communitarian participation and a new position of
science (and of scientists) for the collective search of solutions. Since up to 2008 Holambra
had never tried a so great enterprise in health and environment, our objective were to start
an investigation on the existence of basic conditions for an Ecosystem Approaches to Health
(EAH) compatible with the surveillance and control of schistosomiasis transmission in that city.
Methodology: The inquiry begun in July of 2008 and was based on the identification of the
stakeholders to compose an “Extended Peer Community” aiming at a genuine participative
and transdisciplinary approach. For this, there were considered sectors of the civil society, the
economy and the politics affected by schistosomiasis and/or with presumable influence in
its facing, and the available institutional structure was analyzed. The information was gotten
from documentary analysis (periodicals, SINAN database, acts of sessions of the Chamber of
Councilmen etc.), from historical and technical narratives of the sectors involved in the local
control and surveillance of schistosomiasis and from consultations to the unions and to the
City councils of Health and of Environment. The information had been examined according to
the theory on EAH, with emphasis in its relation with Post-Normal Science. Results: Amongst
the conditions considered favorable to the accomplishment of a EAH we emphasize: 1. The
indispensable role of the Family Health Program (PSF) as interlocutor in the planning and
execution of a diagnostic, treatment and information process, performed from July 2008 to
May 2009, by articulating the city hall sectors to the population and to the state and federal
research institutions. 2. The importance of the Ist Week of Schistosomiasis, state initiative
event carried through in 2009 by the municipal city hall and FIOCRUZ, which stimulated
debates on sanitation, involving some representatives of the civil society. Amongst the
favorable conditions we detach that: 1. In Holambra some important interests related to the
288
problem of schistosomiasis have no representation in governmental institutions nor at the
civil society. For example, the unions are fragile and rarely compromised with the workers
interests, and the institutional structure to deal with the environment is not compatible with
the relevance of the existing problems, including schistosomiasis transmission. 2. After the
municipal elections, in January of 2009, the strong political instability paralyzed and modified
the administrative structure drastically, with severe consequences for the structures of the
PSF and other municipal health sectors. On the other hand, this conflicts stimulated the
debate on local problems and the political participation, what tends to favor the maturation
of conditions for a genuine participation of the population in an EAH. Conclusions: The basic
conditions for an EAH in Holambra are not yet mature and the current political- administrative
instability complicate the planning and the accomplishment of audacious enterprises as the
intended one, which is so dependent on social contract. However, the recent changes in the
political participation patterns represent a promising way to the necessary maturity
289
tREnd Of tHE SCHIStOSOMIaSIS ManSOnI In MaRanHãO StatE: 1997 tO 2003.
Selma Patricia diniz Cantanhede; Inês Echenique Mattos; aldo Pacheco ferreira
Escola Nacional de Saúde Pública, Fiocruz, RJ - Brasil
Introduction: The parasitic disease called schistosomiasis is still considered a major
public health problem. After the malaria parasite is the most prevalent worldwide, with
approximately 200 million infected individuals. In Brazil, it is assumed that there 2.500.000 to
8.000.000 of Schistosomal, northeastern is the most affected area. In the state of Maranhão,
schistosomiasis is reported since 1920. Despite control activities undertaken in this state, it
is noted that the contribution of characteristics for the low socioeconomic development has
given the maintenance of the same disease. In that sense, this work was to analyze the trend
in the percentage of positive cases of schistosomiasis in Maranhao, in the period 1997 to
2003. Methodology: The work consists of an ecological study of time series, corresponding
to the percentage of cases positive for schistosomiasis in the state of Maranhao, grouped by
regional health authorities. We analyzed the 15 regional health information system available
on the Schistosomiasis Control Program (SISPCE): Bacabal, Balsas, Caxias, Chapadinha, Codó,
Colinas, Imperatriz, Pedreiras, Pinheiro, Presidente Dutra, Santa Inês, São João dos Patos,
São Luís, Viana, Vitorino Freire. It made a trend analysis for each regional health service,
using polynomial regression models. The coefficient of determination (R2) and residue
analysis were used to define the best model. They considered the model as statistically
significant when p ≤ 0.05. The development of the polynomial regression was performed
using SPSS for Windows version 17.0. Results: The total percentage of positive cases of
schistosomiasis for the study period amounted to 7.42%. The Regional Health São Luís e
Bacabal showed a downward trend and constant throughout the series. The average annual
percentage of positive cases for these health regions was 1.72 and 0.66, respectively. The
regional health Colinas showed tendency to increase until the year 2002, with a subsequent
decrease in the percentage of cases. At the regional health of Imperatriz trend was observed
an increasing trend, with average annual cases corresponding to 0.41. Conclusions: The
analysis results showed that, due to socioeconomic characteristics, the trend of decline in
the percentage of cases occurring in regional health Bacabal, raises questions and suggests
the possibility of underreporting. Already in the regional health and Imperatriz e Colinas,
issues relating to environmental sanitation and poverty are also significant and certainly
are related to the increasing percentage of positive cases. The decline in the percentage of
cases of schistosomiasis observed in the regional health of São Luís was possibly influenced
by the best living conditions existing in the state capital. It is believed that the registration
of data concerning the health regions of the state of having been affected by the changes
that occurred due to decentralization of the Schistosomiasis Control Program. If this event
occurred, it is important to stress that it was not strong enough to mask the details of
the regional health Colinas and Imperatriz. The results of this study indicate aspects that
transcend the analysis of the pattern of occurrence of cases of schistosomiasis in the state
of Maranhao, getting established, especially the need for improvements related to the living
conditions of the state population.
290
USEfULnESS Of tHE nEtWORK Of COntaCtS fOR tHE dEtECtIOn Of SCHIStOSOMIaSIS
In LOW IntEnSItY tRanSMISSIOn fOCUS, EStEIO, RIO gRandE dO SUL
daniela Martins toniolli; Maria Ceci Salcedo Botelho; José antônio Silveira; Carlos graeff
teixeira;
Pontifícia Universidade Católica do Rio Grande do Sul, RS - Brasil
Introduction: In areas of low intensity transmission, diagnostic tests used in classical
endemic areas do not have the required sensitivity. Application of special methods and the
concentration of efforts in the risk group are essential for the success of control measures.
We report the result of concentrated efforts in a risk group identified from population census
and enlarged by successive additions of information from the detailed epidemiological
investigation in this group. Methodology: Esteio (29 ° 51 ‘41‘‘ S; 51 ° 10‘ 45‘‘ W) is in the
metropolitan region of Porto Alegre, Rio Grande do Sul, southern Brazil. Population residing
in the surroundings of the three transmission foci had demographic and risk assessment
performed at a census in april, 1999. The identified group of risk (history of contact with
the known foci) has been under revision since march 2010. Three total stool samples were
collected from 17 individuals. 30g of each fecal sample was submitted to Helmintex method.
Briefly stools were dissolved in saline, filteres through surgical gauze, washed by spontaneous
sedimentation, sieved, concentrated by Ritchie method and eggs were isolated through
interaction with paramagnetic beads in a magnetic field (Bangs Labs, EUA). Eggs present
in the final sediment were identified in the microscope. Results: Three out of 17 patients
were positive, with estimatives of egg per gram of feces (epg) ranging from 0.03 to 0.2. Two
young children are most probably cases of reinfection and 1 individual is a new diagnosis but
probably not a new infection according to the epidemiological revision along the last years.
Conclusions: The results demonstrate the importance to concentrate efforts for diagnosis of
human cases in the risk group established through constant and prolonged epidemiological
investigations. This approach leads to the establishment of a network of contacts sharing
the risk for the infection. Interruption of transmission, the proposed objective for control
measures in low intensity transmission areas, stems on efforts concentrated with the best
available diagnostic methods on the group of risk instead of whole population approaches.
291
EvaLUatIOn Of tREatMEnt WItH antIBOdIES antI CO-StIMULatORY MOLECULES
On CELLULaR RESPOnSE Of SHIStOSOMIaSIS
Laís Cristina de Souza; naiara naiana dejani; Joice Margareth de almeida Rodolpho; Sandra
Regina Pereira de Oliveira; débora Meira neris; vitor Leão; Ricardo de Oliveira Correia; Carlos
alberto Soares; auro nomizo; Heloísa Sobreiro Selistre de araújo; fernanda de freitas anibal
Universidade Federal de São Carlos, SP - Brasil
Introduction: Schistosomiasis mansoni is a neglected disease, chronic parasite that affects
millions of people around the planet. During infection the individual shows granuloma
formation with cellular infiltrate and can progress to fibrotic tissue. Several studies have
demonstrated the involvement of T cells in this type of response. This inflammatory process
is characterized by a mixed inflammatory reaction with cellular infiltrate composed of
mononuclear cells, polymorphonuclear cells, eosinophils, epithelioid cells and fibroblasts.
The granulomatous process in schistosomiasis may be dependent on CD4 + and requires
recruitment and accumulation of inflammatory cells at the site of deposition of eggs.
Experimental manipulations using MAbs that block the activation of T cells is an interesting
field of intervention in this type of response. The co-stimulatory signals may have different
roles in various types of immune response. Thus, the aim of study was to evaluate the
cellular response in blood leukocytes from animals infected with Schistosoma mansoni
after treatment with MAb anti-CTLA-4. Methodology: We used 24 female Balb / c mice. The
animals were divided in four groups: untreated control (G1) / treated with irrelevant IgG
(G2) and infected without treatment (G3) and treated with MAb anti-CTLA-4 (G4) 7 days
post infection (100ug / kg / animal). After treatment (13 post infection) the animals were
euthanized to obtain blood and subsequent evaluation of the leukocyte response. Results:
Animals infected with S. mansoni showed increase total leukocytes in the blood compared
to control animals without treatment. Infected mice that were treated with MAb decreases
in total leukocytes in the blood compared with animals only infected. Conclusions: According
to the results suggest that treatment with MAb anti-CTLA-4 negatively modulates the influx
of leukocytes in the blood of animals infected with S. mansoni. There were no reports in the
literature on the role of this molecule in the regulation and control of schistosomiasis. Thus,
studies in immune intervention during schistosomiasis may contribute with new options to
improve and expand current therapy. Support: FAPESP
292
ExPERIMEntaL anaLYSIS Of tHE IMIdazOLInE dERIvatIvE 3-BEnzYL-5-(4-CHLOROaRILazO)-4-IMIdazOLIdInE-2-OnE agaInSt adULt WORMS Of SCHISTOSOMA
MAnSOnI
andréa Cristina aplinário Silva; Juliana Kelle de andrade Lemoine neves; andré de Lima aires;
anekecia Lauro da Silva; tiago Moreira alves feitosa; Renata alexandre Ramos Silva; Maria
do Carmo alves de Lima; Ivan da Rocha Pitta; Suely Lins galdino; Mônica Camelo Pessoa de
azevedo albuquerque
Introduction: Schistosomiasis is one of the ten tropical diseases that make part of
control programs of the World Health Organization since 2000. Nevertheless, it is still
considered a neglected disease because there is not a real effort of the pharmaceutical
industry for the development of new effective drugs and control means. The main way
against schistosomiasis is chemotherapy and the only active drug against all species
of Schistosoma is praziquantel. This drug is already used as antischistosomal agent for
over 40 years. However, as consequence of its large and exclusive use in the endemic
areas, a potential danger has been arisen: the emergence of strains of Schistosoma
resistant to praziquantel. This background points to the necessity of the study of
new schistosomicidal drugs. In this context, imidazoline derivatives have presented
promissory results. The present work evaluated the in vivo schistosomicidal activity of
the imidazoline derivative 5-(4-chloro-arilazo)-3-benzyl-4-tioxo-imidazolidine-2-one LPSF/PT05 against adult worms of Schistosoma mansoni. Methodology: Swiss Webster
albino female mice were infected with 80 cercariae of S. mansoni (BH strain) by the
technique of caudal immersion. After 45 days of infection, animals were orally treated
with the synthetic compound LPSF/PT05 during 5 consecutive days in the following
concentrations: 10, 30 and 100 mg/Kg/day. The derivative LPSF/PT05 was prepared
by mean of dispersion with a hydrophilic polymer polyethyleneglycol (PEG). Only PEG
was administered in the animals of the control group. Animals were sacrificed at 15th
day post-treatment. The susceptibility of the adult worms of S. mansoni to the studied
derivative was evaluated through the recovery of worms by mean of the technique of
perfusion of the portal hepatic system and mesentery and of the ovoposition pattern by
the oogram test. ANOVA test was performed to verify any differences among data from
groups (α = 0.05). Results: The average number of recovered adult worms in controls and
in animals treated with 10, 30, and 100 mg/Kg/day was 31.0 (± 2.1), 15.8 (± 7.8), 15.8 (±
5.6), and 11.2 (± 3.5) respectively. Thus, independent of drug concentrations employed,
there was a significant decreasing of the number of worms collected in the animals
subjected to treatment as compared to controls (p < 0.05). In the concentrations of 10
and 30 mg/Kg/day, the remission was approximately 50% as compared to the controls.
For the concentration of 100 mg/Kg/day, the remission was of 64.52%. Oogram test
showed all stages of the eggs independently of studied groups. In the treated animals,
the percentage of mature eggs in the dosage of 10, 30, and 100 mg/Kg/day was of
42.73%, 39.68%, and 44.48% respectively while in controls was of 26.34%. In a similar
way, the analysis of not viable eggs exhibited in the drug concentrations of 10, 30, and
100 mg/Kg/day a percentage of 27.43%, 22.56%, and 29.12% while in controls was of
293
9.23%. Conclusions: The imidazoline derivative LPSF/PT05 exhibited schistosomicidal
activity against adult worms of S. mansoni in the administered concentrations. It was
observed a decreasing of the number of recovered worms and an increasing of mature
and not viable eggs. Thus, we suggest that the presence of all stages of S. mansoni eggs
is probably related to the residual parasitic charge.
294
ExPERIMEntaL CHEMOtHERaPY In SCHIStOSOMIaSIS USIng RUtHEnIUM nItRIC
OxIdE dOnOR
francine nesello; Jean Jerley nogueira da Silva; Carolina Panis; Ivete Conchon Costa; Maria
Claudia noronha dutra de Menezes; francisco José de abreu Oliviera; thiago Mattar Cunha;
Maria angélica Ehara Watanabe; Wander Rogério Pavanelli
Introduction: Schistosomiasis is one of the major health problems in many developing countries
and causes liver damage. The only drug available for treating schistosomiasis is praziquantel,
however there are already reports of resistance to its use in treatment, making it necessary to
search and develop new compounds to combat schistosomiasis. NO donor compounds have
low toxicity in vitro and in vivo and are stable in aqueous media in the presence of oxygen and
NO released by reducing agents that are present in the host inflammatory microenvironment.
These donor compounds have recently emerged as an interesting and important alternative
treatment to experimental several models (Chagas disease, Leishmanioses and Toxoplasmosis).
It is recognize that one of the most prominent functions of NO in the immune system is its
participation in protective immunity, may directly and indirectly modulate the inflammatory/
immune response. We tested, two new products (trans-[Ru(NO)(NH3)4pz] or [Ru(NH3)5NO]
(PF6)3 ruthenium nitrosyls derivatives, which release nitric oxide (NO), in BALB / c mice
infected with the BH strain of Schistosoma mansoni. Methodology: Efficacy and safety of these
compounds were analyzed through of 1- mortality curve and parasite number; 2- inflammatory
response by histopathological characteristics of the granulomatous lesions – (H&E); collagen
deposition-fibrosis (picrosirius red) and immunohistochemistry (IH) staining of Vascular
Endothelial Growth Factor (VEGF) and 3- immune response through of cytokines and nitric
oxide production (ELISA). These parameters were analyzed in two periods (45º and 90º) after
infection. Results: Initially, we found that NO-donor-treated mice are more resistant to infection,
since they exhibited higher survival compared with control group. This result is directly related
to reduction in the number of parasites (adult worms) found in the liver. Furthermore, we
observed a decreased influx of inflammatory cells (granuloma numbers) into the liver tissue of
treated mice in both periods analyzed (45 and 90 days p.i.). Consequently, the collagen deposition
(fibrosis) on liver these animals was reduced. In addition, the amount cells expressing VEGF,
was decreased in the liver of [Ru(NH3)5NO](PF6)3-treated mice in both analyzed periods, than
compared with control group. The ELISA test detected an increased production of TNF-α and
IL-10 in liver of NO-donor-treated mice, but not difference than IL-4 production. In addition,
the amount of NO in serum was higher only in mice treated with (trans-[Ru(NO)(NH3)4pz] than
compared with untreated animals. Our results clearly show that NO-donor treatment resulted
in the kill parasites (parasite multiplication), amelioration of the inflammatory response in the
liver, with reduction of hepatic damage and significantly promoted an increase in the survival
these mice. Conclusions: In conclusion, this work showed that the compound acts in the
modulating of the immune response against the parasite, regulation the influx of inflammatory
cells, consequently promoted a clearance of parasite burden. Therefore, administration
of NO donors and other drugs in conjunction can constitute a promissory therapeutic
avenue that could be explored as a new alternative for the treatment of schistosomiasis.
295
ExPERIMEntaL StUdY Of SCHISTOSOMA MAnSOnI ISOLatE fROM PatIEnt WItH
tHERaPEUtIC faILURE tO PRazIQUantEL
Maria José Conceição; Eric vinaud de Melo; aline Eduardo Carlôto; natália Machado da Silva;
Iran Mendonça da Silva; Isabel Cristina Melo Mendes; viviane Pereira; nelson gonçalves
Pereira; José Borges-Pereira
UFRJ-Faculdade de Medicina, Fiocruz, RJ - Brasil
Introduction: Patients infected with ´Schistosoma mansoni´ and assisted in the outpatients
clinic of the Service of Infectious and Parasitic Diseases of Hospital Universitário Clementino
Fraga Filho-Universidade Federal do Rio de Janeiro-UFRJ have responded to treatment with
Praziquantel, which was initiated in the 70’s decade. Failure of successive treatments with
Praziquantel observed in one patient has roused an experimental study. Methodology: CASE
REPORT: Male patient of 42 years old, born in Bahia state, with hepatointestinal form, positive
for´S. mansoni´ according to Lutz’s method and Kato’s, modified by Katz et al., treated with
Praziquantel, single oral dose, first two doses with 40 mg/kg weight, and the third with 60
mg/kg. Control exams were done after 60, 90 and 180 days of treatment and rectal biopsy,
after 180 days. EXPERIMENTAL STUDY: The isolates of ´S. mansoni´ were kept in groups of
thirty ´Biomphalaria glabrata´ (8 to 10 miracidia for each specimen- Chaia´s method) in the
three treatment phases. After 45 to 60 days, cercariae (50) were inoculated by percutaneous
route, individually, into groups of 60 Swiss webster mice. Fifty days later, the animals were
treated with Praziquantel-200 mg /kg, by oral route, and after 14 days, they were submitted
to perfusion of portal system. Half of the animals has not been treated. It was maintained
a control group, which members were inoculated with ´S. mansoni´ of patients that had
responded to Praziquantel treatment. Results: CLINICAL STUDY- Parasitological stool tests
that were done in the patient after 60, 90 and 180 days of treatment, and rectal biopsy
after 180 days in the three stages of Praziquantel treatment revealed parasite eggs expelling.
EXPERIMENTAL STUDY- Swiss webster mice that were treated with Praziquantel and
submitted to perfusion of portal system and mesentery in the three stages of experimental
study have presented alive worms in all the groups. Conclusions: The non-responsiveness
of the patient to three successive treatments with Praziquantel, rejecting the possibility of
reinfection, associated to experimental demonstration of therapeutic failure, emphasizes the
necessity of promoting more researches on new drugs that ensure the effective treatment in
patients with schistosomiasis.
296
IMPROvIng MEtHOdS Of IndIRECt ELISa USIng antI-Igg antIgEn SOLUBLE adULt
WORM (SWaP), SOLUBLE Egg antIgEn (SEa) and tEgUMEnt Of SCHIStOSOMULa
antIgEn (SMtEg) In SERa Of PatIEntS tHE CHROnIC PHaSE Of SCHIStOSOMIaSIS
ManSOnI
Watson Herman Martins; Rafaella fortini; Elizandra giani Ribeiro; vanessa Silva Moraes;
Edward Jose de Oliveira; Paulo Marcos zech Coelho
Introduction: Currently, the most widely used method for laboratory diagnosis of
schistosomiasis is the search of parasite eggs in fecal samples, demonstrated mainly by
the Kato-Katz technique. However, one important limitation is the variation of sensitivity
according with the amount of slides, number of eggs released and number of samples.
The simultaneous use of diagnostic tests such as examination of feces and detection of
antibodies has been used to increase sensitivity and specificity of diagnosis. The serological
techniques has potential for diagnosis of schistosomiasis (mainly those that detect IgG antibody produced in large quantities at the infection), but this is not yet part of the routine
of clinical laboratory. This is due to standardization and validation of the technique itself,
targeting a specific and sensitive diagnosis of the disease especially in patients with low
parasitic burden. Methodology: We collected serum samples of patients living in an endemic
areas (Penha do Cassiano, MG Brazil), previously diagnosed by the Kato-Katz method.
Serum samples from were collected individuals adults and certainly note infected. These
were validated in comparison with serum of neonates. Indirect ELISA anti-IgG was carried
out with human samples, using plates coated with SEA (soluble egg antigen of S. mansoni)
SWAP (soluble adult worm antigen) and SmTeg (coat antigen of schistosomula). The
standardization was made by the representative curve at different dilutions and with covered
plate at different antigen concentrations. Serum samples were used in all experiments for
positive and negative controls. The results were obtained in the microplate reader at 450 nm.
Data obtained by absorbance values were analyzed by Minitab software, using the normality
test Kolmogorov-Smirnov normality and the Students t test (p> 0.05) for analysis between
groups. The sensitivity, specificity, and cut off likelihood ratio were determined by Prism 4.0
software. Results: When comparing the results obtained by the methods of SWAP- ELISA
and ELISA-SEA-ELISA Smteg was observed that there was a positive relationship between
parasite load, previously diagnosed by Kato- Katz and the absorbance value in both tests.
Sensitivity values and specificity of each diagnostic technique for human diagnosis were
respectively: 100% and 97% (cut off = 0189) for SWAP-ELISA, 75% and 75% for SEA-ELISA
(cut off = 0311) and SmTeg-ELISA (cut off = 0318). We observed a direct correlation between
eggs per gram of feces (epg) and titles of the tests. Conclusions: Thus, the three techniques
presented excellent performance. Using the SWAP-ELISA showed better efficiency, when
compared with the two other techniques using human serum. Keywords: Schistosomiasis
mansoni, serodiagnosis, ELISA.
297
In vItRO SCHIStOSOMICIdaL aCtIvItY Of (-)-6,6’-dInItROHInOKInIn agaInSt
SCHISTOSOMA MAnSOnI
Lizandra guidi Magalhães; ana Carolina Pereira; thais Coelho Lima; Karen C S Rezende; Jairo K
Bastos; Márcio L a e Silva; vanderlei Rodrigues; ademar alves da Silva filho; Wilson Roberto
Cunha
Introduction: Schistosomiasis, a chronic debilitating disease that affects 200 million people in
tropical and subtropical regions causing more than 280,000 deaths per year. Reduced cure
rates and the failure of treatment after praziquantel administration have been shown in
patients. In addition, the existence of resistant strains reinforces the need to develop new safe
and effective schistosomicidal drugs. In the last decade, there has been intensification in the
search of antiparasitic compounds from natural sources, mainly from plants, which continue
to be a major source of biologically active metabolites that may provide lead structures for
the development of new drugs. (-)-6,6’-dinitrohinokinin belong to the dibenzylbutyrolactone
lignan class of compounds, which is among the natural products of interest, since many of its
compounds display a broad range of biological activities with therapeutic potential. Therefore,
the aim of this work was to evaluate in vitro schistosomicidal activity of (-)-6,6´dinitrohinokinin,
a semi-synthetic derivative of the natural lignan (-)-hinokinin isolated from dry seeds of Piper
cubeba Methodology: (-)-6,6’-dinitrohinokin was obtained by nitration of the aromatic rings of
compound (-)-hinokinin, as previously described by Souza et al, (2005). For schistosomicidal
assay , (-)-6,6’-dinitrohinokin was dissolved in DMSO and used in concentrations varying from
10 to 200 μM, which were added to the medium containing one adult worm pair after a period
of 24 h of adaptation to the culture medium. The parasites were kept for 120 h and monitored
every 24 h to evaluate their general condition: motor activity, alterations in the tegument, and
mortality rate. Also, changes in the pairing, egg production, egg development were evaluated
using an inverted microscope, and the viability assay was performed as described by Comely et
al. (1989). The negative control worms were treated with 1% DMSO in an RPMI 1640 medium
and positive control worms were treated with 10 μM,praziquantel. Results: The results shows
that after 24h of incubation with (-)-6,6’-dinitrohinokin (25-200μM), 100% the adult worms pairs
separated into individual male and female and was observed that incubation at concentrations
100 and 200μM, 50 and 100% adult worms were dead, respectively. In addition, worms
showed decreased motor activity and tegumental alterations when compared with control
groups. Moreover, the results showed that (-)-6,6’-dinitrohinokin decreased the worm viability
of parasites producing a dose–response effect in comparison with the negative control group.
Moreover, (-)-6,6’-dinitrohinokin (25 to 100 μM) inhibited the egg production in comparison
with the negative control group. However, this effect was observed as the result of the separation
of the worms by the action of (-)-6,6’-dinitrohinokin. On the other hand, at concentration 10
μM wasn´t shown a significant decrease in the number of eggs after 120 h of incubation. Also,
(-)-6,6’-dinitrohinokin (50 and 200 μM) affected the development of eggs produced. Conclusions:
In summary, our results indicate that (-)-6,6’-dinitrohinokin possesses in vitro schistosomicidal
activity against S. mansoni. It is also important to emphasize that, considering the obtained
results, further biological studies are in progress in order to elucidate its effects in vivo,
cytotoxicity, and its mechanism(s) of schistosomicidal action. Financial Support: FAPESP, CNPq
298
In vIvO EvaLUatIOn Of tHE IMIdazOLInE dERIvatIvE 3-BEnzYL-5-(4-CHLOROaRILazO)-4-IMIdazOLIdInE-2-OnE agaInSt EvOLUtIvE fORMS Of SCHISTOSOMA
MAnSOnI
amanda Caroline Malaquias de farias; Juliana Kelle de andrade Lemoine neves; andré de
Lima aires; tiago Moreira alves feitosa; Renata alexandre Ramos da Silva; Maria do Carmo
alves de Lima; Ivan da Rocha Pitta; Suely Lins galdino; Mônica Camelo Pessoa de azevedo
albuquerque
UFPE, PE - Brasil
Introduction: In last 40 years, praziquantel (PZQ) improved the therapy against the
schistosomotic infection significantly. The parasitological cure with the use of PZQ
depends on time of infection, on strain and sex of worms and if they are matched or not,
and on drug dosage. Unfortunately, higher disadvantage of PZQ is its low efficacy against
immature forms of Schistosoma. Several alternatives have been proposed to solve this
problem such as the association of PZQ with other drugs and the continuous research for
new antischistosomal drugs acting upon the younger forms of S. mansoni. In this context,
imidazolidine derivatives have presented potential schistosomicidal activity against adult
worms of S. mansoni in in vivo and in vitro studies. The present work aimed to evaluate
the action of the derivative 3-benzyl-5-(4-chloro-arilazo)-4-imidazolidine-2-one (LPSF/PT05) in mice infected by Schistosoma mansoni considering the following worm evolutional
forms: skin schistosomule, lung schistosomule and juvenile worms in the portal-hepatic
system. Methodology: Thirty-day-age female mice weighing approximately 30 and 36 g
were percutaneously infected with 50 cercariae of S. mansoni (BH strain). After infection,
animals were randomly divided in three experimental groups. Composition of these
groups was based on following phases of worm development: skin schistosomule (G1),
lung schistosomule (G2) and juvenile worms in portal-hepatic system (G3). Each group was
formed by two subgroups each one containing ten animals: one subgroup treated with
LPSF/PT05 and another receiving only the drug vehicle (control group). Therapy occurred
during five consecutive days. Mice of G1, G2, and G3 groups were treated with 100, 30 and
100 mg/Kg/day of LPSF/PT05 respectively. The experimental intervention of G1, G2 and
G3 animals was performed at 1st to 5th, 14th to 18th and 30th to 34th days after infection
respectively. All animals were sacrificed fifty days after infection. The susceptibility of the
worms of S. mansoni to the studied derivative was evaluated through the recovery of
worms by mean of the technique of perfusion of the portal hepatic system and mesentery,
of the ovoposition pattern by the oogram test and the elimination of eggs in the stool by
Kato-Katz method. ANOVA test was performed to verify any differences among data from
groups (α = 0.05). Results: According to statistical analysis, it was not observed difference
(p > 0.05) with respect to the number of adult worms recovered in different studied
groups. The average number of worms recovered in G1-control, G1-treated, G2-control,
G2-treated, G3-controle, and G3-treated was 13.7 (± 1.7), 12 (± 4.0), 11.4 (± 7.9), 12.3 (±
1.5), 7.3 (± 1.1), and 7.0 (± 2.0) respectively. With respect to therapeutic scheme employed
in this study, the derivative LPST/PT05 did not alter the ovoposition pattern significantly.
Additionally, analyses of intestinal fragments showed all developmental stages of eggs
with similar percentages for all studied groups and the number of eggs released per gram
299
of stool did not also exhibit significant alterations when different groups were compared.
Conclusions: In vitro studies have demonstrated that the derivative LPSF/PT05 present
schistosomicidal activity against adult worms of S. mansoni. However, considering the
therapeutic scheme adopted for this study, LPSF/PT05 did not exhibit in vivo activity
against S. mansoni in the evolutional phases of skin schistosomule, lung schistosomule,
and juvenile worms in the portal-hepatic system.
300
In vIvO EvaLUatIOn Of tHE IMIdazOLInE dERIvatIvE LPSf/RzS-5 agaInSt adULt
WORMS Of SCHISTOSOMA MAnSOnI
Juliana Kelle de andrade Lemoine neves; andré de Lima aires; anekécia Lauro da Silva; tiago
Moreira alves feitosa; Renata alexandre Ramos Silva; Maria do Carmo alves de Lima; Ivan da
Rocha Pitta; Suely Lins galdino; Mônica Camelo Pessoa de azevedo albuquerque
Introduction: Schistosomiasis is the second most important human parasitic disease with
regard to mortality. The global estimate is approximately 300 million infected individuals
distributed in 76 countries of whom 20 million develop serious complications, causing
about 500,000 deaths per year. In Brazil, this disease exclusively is caused by the species
Schistosoma mansoni with estimate of more than 14 million of infected individuals and,
among the Brazilian States, Pernambuco presents the higher prevalence of infected
persons. Praziquantel is the first choice drug against S. mansoni but several reports relate
the emergence of resistance for this drug. For this reason, it is very important to find new
drugs presenting schistosomicidal activity, such as the imidazolines that are a large class
of bioactive compounds and derivatives exhibiting schistosomicidal properties. However,
few studies have been performed about this subject. Present work evaluated the in vivo
schistosomicidal activity of the imidazoline derivative LPSF/RZS-5 against adult worms of
Schistosoma mansoni (BH strain). Methodology: Swiss Webster albino female mice were
infected with 80 cercariae of S. mansoni (BH strain) by the technique of caudal immersion.
After 50 days of infection, animals were orally treated with the synthetic compound LPSF/
RZS-5 (50 mg/Kg/day) during 5 consecutive days. Controls received only the vehicle of the
drug. At 49th days of infection (pretreatment phase) and at the 6th and 14th days posttreatment, mice were analyzed with respect to the elimination of eggs in the stool by KatoKatz method. At 14th day after-treatment, animals were sacrificed and intestinal fragments
were microscopically analyzed by the oogram test. ANOVA test was performed to verify any
differences among data from groups (α = 0.05). Results: In the present study, the derivative
LPSF/RZS-5 showed in the group of infected and treated animals a decreasing in the number
of eggs in the stool at the 14th day post-treatment (318 Eggs Per Gram - EPG) as compared to
the pretreatment phase (629 EPG) in the same group of animals (p < 0.05). In controls, it was
not observed any alterations in the eggs counts in the different studied phases. Moreover, at
the 14th day, the decreasing percentage of EPG was 62.1% for the treated (318 EPG) animals
as compared to controls (512 EPG). Oogram test exhibited an increasing in the percentage of
not viable eggs in the intestinal fragments of treated mice (23.8%) as compared to controls
(8.6%), but all phases of egg-development were visualized. Conclusions: Although in vitro
analysis the LPSF/RZS-5 has been presented 100% of mortality upon adult worms of S.
mansoni, in the present work, this derivative did not show in vivo significant activity against
these worms in the drug concentrations utilized. However, the pattern of ovoposition that
was found in this work suggests a small and temporary interference on posture of eggs. More
studies with different concentrations and therapeutic schemes of the LPSF/RZS-5 have been
developed in attempt to better understand its probable antischistosomal property.
301
In vIvO SCHIStOSOMICIdaL EffECt Of tHE IMIdazOLInE dERIvatIvE 5-(4-fLUORaRILazO)-3-BEnzYL-4-tIOxO-IMIdazOLIdInE-2-OnE agaInSt adULt WORMS Of
SCHISTOSOMA MAnSOnI
Juliana Kelle de andrade Lemoine neves1,2; andré de Lima aires2,3; tiago Moreira alves
feitosa2; anekecia Lauro da Silva1; Renata alexandre Ramos Silva2; Maria do Carmo alves
de Lima1; Ivan da Rocha Pitta1; Suely Lins galdino1; Mônica Camelo Pessoa de azevedo
albuquerque
Introduction: Schistosomiasis is a parasitosis caused by species of helminths, trematodes,
whose definitive host is the man. Severity and organic deficit produced by schistosomiasis
in some cases become it the second most important tropical disease in terms of morbidity
and mortality. This disease is distributed in 76 countries where there are approximately 300
million of infected individuals, 600 million live in infection risk areas and about of 200,000
persons die per year. Lack of vaccine and effective actions of public health contribute
to become the chemotherapy the only immediate way to subside the prevalence and
incidence of schistosomiasis globally and praziquantel is the only drug recommended
by World Health Organization for its treatment. However, praziquantel is being used for
over 40 years and thus it has been observed the emergence of praziquantel-resistant
strain of S. mansoni. Some alternatives have been proposed for solve this problem such
as the association of praziquantel to other drugs or the development of new drugs.
Inside this purpose, imidazolines are a large class of bioactive compounds presenting
schistosomicidal properties against adult worms of S. mansoni that was demonstrated in in
vitro studies. Present study analyzed the in vivo susceptibility of S. mansoni (BH strain) to
the imidazoline derivative 5-(4-fluor-arilazo)-3-benzyl-4-tioxo-imidazolidine-2-one (LPSF/
PT09). Methodology: Swiss Webster albino female mice were infected with 80 cercariae
of S. mansoni (BH strain) by the technique of caudal immersion. After 45 days of infection,
animals were orally treated with the synthetic derivative LPSF/PT09 during 5 consecutive
days in the following concentrations: 100 and 150 mg/Kg/day. This derivative was prepared
by mean of dispersion with a hydrophilic polymer polyethyleneglycol (PEG). Only PEG was
administered in the animals of the control group. All animals were sacrificed at 15th day
post-treatment. The susceptibility of the adult worms of S. mansoni to the LPSF/PT09 was
assessed through the recovery of worms by mean of the technique of perfusion of the
portal hepatic system and mesentery and of the ovoposition pattern by the oogram test.
ANOVA test was performed to verify any differences among data from groups (α = 0.05).
Results: The average number of adult worms of S. mansoni recovered in control animals
was 30.0 (± 3.5) while in those treated with LPSF/PT09 was 7.2 (± 2.4) and 14.1 (± 5.4) in
the drug concentrations of 100 and 150 mg/Kg/day respectively. Thus, as compared to
controls it was observed to the concentrations of 100 and 150 mg/Kg/day a decreasing
of 76.67% and 53.34% respectively. Oogram test exhibited worm eggs within all stages
of development for both studied concentrations but the percentage number of mature
and not viable eggs were higher independently of tested group. However, this increasing
it was not significant (p > 0.05). Conclusions: The use of imidazoline derivative LPSF/PT09
in different concentrations showed significant schistosomicidal activity but when it was
302
administered at 150 mg/Kg/day its effect was higher. In spite of the reduction of worm
number observed in treated animals, the oogram test showed similar results to controls
due to the residual parasitic load presented by the treated groups.
303
InHIBItORY EffECtS Of HARPAGOPHYTuM PROCuMBEnS On dISPOSaL Of EggS
dURIng InfECtIOn WItH SCHISTOSOMA MAnSOnI
Joice Margareth de almeida Rodolpho; Sandra Regina Pereira de Oliveira; Laís Cristina de
Souza; naiara naiana dejani; débora Meira neris; Ricardo de Oliveira Correia; Jame‘s almada
da Silva; Paulo Cesar vieira; fernanda de freitas anibal
UFSCAR, SP - Brasil
Introduction: Schistosomiasis mansoni is a neglected disease, chronic parasite that affects
millions of people around the planet. The drug currently used does not prevent re-infection
in endemic regions. Thus, there is the need for repeated treatments, and also encourage the
development of resistant strains and have toxic effects in individuals. One of the paths in
the search for new therapies to combat schistosomiasis is the study of natural substances,
like plants. Harpagophytum procumbens, commonly known as Devil‘s Claw is a medicinal
plant originated from Africa, specifically, the Kalahari Desert, used by natives in the form
of infusion for the treatment of rheumatic diseases, diabetes, atherosclerosis, and malaria.
Pharmacological properties have their active components such as iridoid glycosides 0.5 to
3% that have effects: anti-inflammatory and analgesic and also stimulate the release of
interleukins and leukocyte migration to inflamed. The aim of this study was to evaluate the
inhibitory effect of crude extract of H. procumbens in the elimination of eggs during infection
by S. mansoni. Methodology: Mice were divided into control group, the infected group and
infected group and treated, the treatment was given by gavage for 48 consecutive days with
the following concentrations of crude extract: 15 mg / kg, 30 mg / kg and 60 mg / kg / animal.
After treatment we evaluated the amount of eggs excreted in the feces of infected mice /
treated. Feces were recovered in 48 days after infection / treatment by the method KatoKatz. Results: Results showed a significant reduction of eggs in the group treated with 15 mg
/ kg of crude extract. Conclusions: Thus, our data suggest that treatment with H. procumbens
at a dose of 15mg/kg may contribute to the reduction of eggs during the infection with S.
mansoni, may be a possible alternative for the treatment of schistosomiasis mansoni.
304
OCCURREnCE Of SCHIStOSOMIaSIS In tHE MUnICIPaLItIES Of SãO BEntO and SãO
LUíS, MaRanHãO StatE, In tHE PERIOd 1998 tO 2008
Selma Patrícia diniz Cantanhede; andiara garcez de Souza Silva; Luciana Patrícia Lima alves;
Marjane Soares ferreira; nêuton Silva-Souza
Fiocruz Escola Nacional de Saúde Pública, RJ - Brasil
Introduction: Schistosomiasis mansoni is an important parasitic infection in Brazil. In
Maranhão, the notification of the first cases were recorded in 1920 and is distributed in two
distinct geographic areas, the focus area and endemic areas. The disease occurs in 48 of the
217 municipalities in that state, 26 counties in the focus area and 22 municipalities in the
endemic area, the latter being more specifically located in Baixada Ocidental Maranhense.
With respect to these two geographic areas, must mention the cities of São Bento and
São Luís. The city of São Bento is one of the oldest and poorest state and is located in the
microregion of lower Maranhão State, an endemic area of schistosomiasis. The population
of this town develops hunting, subsistence agriculture and livestock farming, small, and
the main activity is the fishing. Thus, schistosomiasis presents itself as an occupational
disease in this microregion and, consequently, in São Bento. The city of São Luís is the
capital of the state of Maranhão, is located in the focus area and has several neighborhoods
with a high number of cases of schistosomiasis. In the period 1998 to 2004, the districts
Coroadinho and Barreto presented the highest number of cases for this disease, 428 and
287, respectively. The process of occupation of neighborhoods, influenced by migrations
originated mostly from the region of lower Maranhão State is considered one of the causes
of this disease occurring in the county. Considering the importance of the occurrence of
schistosomiasis mansoni in the state of Maranhão and, in particular in the municipalities
of São Bento and São Luís, the present study aimed to survey the percentage of positive
for this parasite in these municipalities, from 1998 to 2008. Methodology: We conducted
a inquiry of data regarding coproparasitologic inquiry conducted in the counties of São
Bento and São Luís, state of Maranhão. The data represent the percentage of positive tests
for schistosomiasis in the period 1998 to 2008 and were supplied by the National Health
Foundation of Maranhão (FUNASA / MA). The coproparasitologic inquiry was conducted by
a team of trained health workers who were responsible for collecting the fecal samples, and
also by laboratory technicians responsible for performing the analysis method by Kato-Katz.
Results: In the survey of archives FUNASA/MA, it was observed that in ten years, 49.152
tests were performed to São Bento and São Luís for 325.955. The percentage of positivity
for schistosomiasis of the population worked was 9.66% in São Bento and 0, 99% in São
Luís during the study period, the city of São Bento had the highest percentage of positivity
for schistosomiasis. In 1999, registered the highest percentage of carriers of the disease,
being 20.1%. Already in 2008, it was found that the lowest percentage corresponded to
6.8%. For the year 2001, no records of activities coproscopic in the county. In São Luís, the
highest percentage is recorded for the year 2008, being 3%. While the lowest percentage
was in 2006, 0.2%. It is noteworthy that the number of tests performed during these ten
years, for these two counties was different. This factor emphasizes the concern with the
city of São Bento, since he submitted a lesser amount of tests and a higher percentage of
patients for the disease. Moreover, São Bento has socioeconomic characteristics that reflect
305
poverty in terms of quality of life of local people. Moreover, although São Luís suburbs
present constituted by irregular occupations, one must consider that this city is the capital
of the state where possibly concentrate the best living conditions of the state. Conclusions:
According to the data obtained, one can see that the cities of São Luís and São Bento are to
sites important in the transmission of schistosomiasis in the state of Maranhão, especially
the city of São Bento, since it found the highest percentages of positivity in the years of study.
These findings indicate the need for public policies to minimize the factors that provide for
the establishment and maintenance of schistosomiasis in these municipalities.
306
PCR aSSaY Of HUMan URInE SaMPLES fOR tHE dIagnOSIS Of SCHIStOSOMIaSIS
ManSOnI
nilton Barnabé Rodrigues; guilherme Oliveira e Silva; Martin J. Enk
Centro de Pesquisas Rene Rachou, MG - Brasil
Introduction: The development of more sensitive and specific diagnostics tests is one of the
priority needs for the control of schistosomiasis mansoni in situations of low prevalence
or low transmission. In this study, a simple and cheap salting out and resin DNA extraction
method for human urine samples was developed, evaluated and applied in PCR assays as
a new molecular tool for the detection of Schistosoma mansoni DNA. Methodology: The
analytical sensitivity, repeatability and reproducibility were determined using serial dilutions
of genomic DNA extracted S. mansoni adult worms. In order to determine the analytical
sensitivity of the PCR assay, humane urine samples artificially contaminated with S.
mansoni DNA in decreasing concentrations were tested. The DNA from these preparations
was extracted using a salting out and resin methodology described recently and amplified
with primers targeting the 121 bp tandem repeat DNA sequence of S. mansoni, previously
designed by Pontes. Results: In a pilot study the reproducibility of the technique reached
100%, showing positive results in 5 assay repetitions of 5 tested samples each containing
20ngDNA/5ml. The efficiency of the extraction procedure was evaluated in a serial dilution of
the original 20ng DNA/5ml sample. Detectable DNA was extracted at a lowest level of 1.28pg
DNA/mL, revealing the high performance of this procedure. Conclusions: The results clearly
demonstrate the efficiency of the extraction procedure. The advantage of this innovative
method is that no toxic substances such as organic solvents or expensive kits are necessary
to successfully perform the extraction. The PCR assay of human urine samples constitutes an
improved and alternative tool for the diagnosis of S. mansoni infections with the potential to
identify individuals with very low infection intensity. Another important future perspective
of the detection of S. mansoni trans-renal DNA is the possibility of early diagnosis of this
disease, considering the probable presence of cell-free, and consequently trans-renal DNA
during the larval stages of human infection. Furthermore the described method offers
operational advantages such as easy collecting and handling of the urine samples under field
conditions as well as examining of large number samples in a short period of time.
307
PORtaL HYPERtEnSIOn In SCHIStOSOMIaSIS ManSOnI: COMPLICatIOnS Of ELECtIvE
SURgICaL tREatMEnt
Izabela voieta; andy Petroianu; vivian Resende; Kelly Rodrigues; vitor freitas fontes; Juliana
Papatella; Carlos Mauricio de figueiredo antunes; José Roberto Lambertucci
Faculdade de Medicina UFMG, MG - Brasil
Introduction: Increased resistance to portal blood flow is the primary factor in the
pathophysiology of portal hypertension; is mainly determined by the morphological changes
occurring in chronic liver diseases. Up to 10% of patients with hepatosplenic schistosomiasis
will eventually develop portal hypertension and esophagogastric varices. Bleeding from
ruptured esophagogastric varices is the most severe complication of schistosomiasis and
is the death cause in a significant group of patients. Pharmacological agents, endoscopic
treatment or surgery can be used to prevent esophageal hemorrhage. Surgical treatment
is indicated in the following situations: 1) documented variceal hemorrhage to prevent
rebleeding; 2) persistent anemia caused by hypersplenism; 3) voluminous spleens causing
distress to the patient; 4) patients with portal hypertension and huge esophagogastric
varices living in rural areas without access to hospitals able to perform complicated
endoscopic or surgical intervention. Surgery, on the course of upper digestive bleeding,
is presently avoided by most hospitals in Brazil due to increased mortality. More recently,
the initial procedure is to interrupt by endoscopic treatment (preferably banding ligation,
sclerotherapy and mechanical compression with a Sengstaken-Blakemore balloon); time is
given for the patient to recover from anemia, dehydration, shock and other intercurrences.
Elective surgery in then performed. This study was set up to evaluate complications of
surgical treatment of patients with schistosomal portal hypertension (total and subtotal
splenectomy, esophagogastric disconnection and suture of gastric varices). Methodology:
Thirty patients attending the outpatient clinic of a University Hospital of Belo Horizonte
(Universidade Federal de Minas Gerais), Santa Casa de Misericórdia de Belo Horizonte
with hepatosplenic schistosomiasis, were selected for this study. They were operated
upon for portal hypertension, from June 2006 to March 2010. Refractory anemia, repeated
episodes of variceal bleeding and huge spleens, were the causes for surgical treatment.
The patients were thoroughly examined during surgery (operative time), up to seven
days postoperative period (mediate) and from the eighth day onwards (late operative).
Results: The age ranged from 19 to 54 years (mean and median: 38) and 23 were male
(77%). Fourteen (47%) and sixteen (53%) out of 30 patients were submitted to total or
partial splenectomy, respectively. Age was not significantly different comparing the two
surgical groups. In the operative time no complications were found. In the mediate time
only minor complications were reported such as, transient fever, discrete wound bleeding
and partial portal vein thrombosis, except for one patient who presented spleen infarction,
abdominal hematoma and fever but recovered with conservative treatment. In the late
operative period the following complications were noted in 3 patients (10%): prolonged
hospitalization, septic shock, pseudohyperkalemia associated with portal vein thrombosis,
hemorrhage; nine months after surgery, one patient evolved with haematemesis, liver
failure, ascites and bacterial peritonitis which culminated with his death. Conclusions:
Except for the patient who died 9 months after surgery in a remote rural area without
308
adequate medical care, the self limited complications above mentioned do not counter
indicate the surgical treatment of portal hypertension in schistosomiasis. To evaluate the
impact of surgery in the life quality of hepatosplenics, a control group of non operated
patients should be equally followed for comparison. This is presently being considered.
309
PRELIMInaRY StUdY Of PULMOnaRY fUntIOnS tEStS In PatIEntS WItH ManSOnI’S
SCHIStOSOMIaSIS
Rita de Cassia dos Santos ferreira; amanda Medeiros gomes da Silva; ana Lúcia Coutinho
domingues; angela Pontes Bandeira
Introduction: Several pulmonary alterations can occur during the course of chronic
schistosomal disease, like pulmonary hypertension and hepatopulmonary syndrome.
However it is possible that minor functional alterations can occur due to organomegaly
or portal hypertension in the hepatosplenic disease. Splenectomy performed in endemic
areas of schistosomiasis aimed to secondary prevention of the hemorrhage from esophageal
varice could reverse this indirect effect in pulmonary function. The purpose of this study was
to compare the spirometric values in patients with hepatosplenic disease and schistosomotic
patients submitted to splenectomy in the past. Methodology: Forty seven patients in
treatment at the schistosomiasis outpatient clinic of the hospital of Universidade Federal
de Pernambuco (UFPE) were enrolled in this study, between march and July 2010. Twenty
five had the hepatosplenic form of the disease and 22 were splenectomized due to prior
hemorrhage from esophagic varice. The study had approval from the Ethics Committee of
the UFPE Center of Health Sciences and written informed consent was obtained from each
patient enrolled. The patients were submitted to spirometry, using the One Flow FVC Kit®,
for determination of forced vital capacity (FVC), forced expired volume in the first second
(FEV1) and FEV1/FVC ratio, selecting at least three maneuvers for analysis. Results: In the
hepatosplenic group the mean percentual values of FVC and FEV1 were 84,61%±13,28%
(CI 95%: 79,40% – 89,82%) and 82,04% ± 15,30% (Cl 95%: 76,04 – 88,04), respectively. The
mean percentual values of FVC and FEV1 in the group of splenectomized patients were
91,54%±11,58% (CI 95%: 86,70%±96,38%) and 87,45% ± 12,17% (CI 95%: 82,36% -92,54%),
respectively . In the hepatosplenic group, 48% of the patients presented with functional
disturbances: 28% had mix or restrictive pattern and 2% obstructive pattern. Thirty two of
the splenectomized patients had functional disturbance: 18% mix or restrictive pattern and
13% obstructive pattern (p= 0,40). Conclusions: The overall mean percentual values of FVC
and FEV1 were normal and without differences among the groups of hepatosplenic and
splenectomized patients, demonstrated by the superimposed confidence intervals of these
variables. However, 48% of the hepatosplenic patients presented with functional disturbances
compared to 32% of functional disturbances among splenectomized patients. Althoug
no statistical difference was found (p≥ 0,05), there was a trend toward more ventilation
disturbance in the hepatosplenic group. The authors speculated that the splenomegaly
or portal hypertension exerts some effect in the functional respiratory capacity of these
individuals
310
PROfILE Of SPECIfIC HUMORaL RESPOnSES agaInSt SCHIStOSOMIaSIS In an
EndEMIC RURaL aREa Of MInaS gERaIS StatE, BRazIL: a LOngItUdInaL StUdY.
Leonardo ferreira Matoso; Ricardo toshio fujiwara; Humberto ferreira de Oliveira Quites;
Lorena Junia de Souza Santos; Luciana Lisboa Mota e Castro; Mery natali Silva abreu; Helmut
Kloos; Rodrigo Correa-Oliveira; andréa gazzinelli
Introduction: Laboratory diagnosis of schistosomiasis is usually performed by direct methods
which detect the parasite eggs. Alternative methods include indirect tests using clinical and/
or serological evaluations. Both methods present limitations in low prevalence areas or
where mass treatment programs are conducted. In this context the impact of chemotherapy
in endemic areas at mid and long terms, due to re-infection, constitute one of the main
problems. Therefore there is a need to explore new avenues that will evaluate the use of
specific immunological markers to predict susceptibility/resistance to human schistosomiasis.
This study aimed to examine longitudinally the pattern of specific IgE and IgG4 against S.
mansoni antigens (SEA or SWAP) of residents of an endemic area of the municipality of
Ponto dos Volantes in the State of Minas Gerais, Brazil. Methodology: Parasitological and
serological analyses were performed in 152 individuals who participated in the entire period
of evaluation. The follow up consisted of collection of three serum and fecal samples (time
zero, one and four years after treatment), in the period between 2001 and 2005. Three stool
samples were collected using the Kato-Katz method. Levels of IgE and IgG4 (SEA/SWAP) were
evaluated by ELISA using sera collected from the enrolled individuals. Sperman’s test was
used to evaluate the correlation between infection intensity and specific levels of IgE and
IgG4 against S. mansoni antigens (SEA or SWAP). Results: Before treatment, the prevalence of
S. mansoni was 57.9% (CI95% 50.06-65.74) and the geometric mean egg counts 64.1 (CI95%
52.67-75.51). After treatment, the prevalence decreased significantly to 15.1% (CI95%
9.41-20.79) and 27.6% (CI95% 20.50-34.70) in 2002 and 2006, respectively. The intensity
of infection was significantly reduced to 37.8 (CI95% 35.07-40.55) and 36.4 (CI95% 33.9338.89) eggs/g after treatment. Analysis of IgE-SEA demonstrated that significant differences
in antibody production was only detected after three years of treatment, with infected
individuals presenting higher anti-SEA IgE levels when compared to those egg-negative.
Although differences in anti-SWAP IgE production were observed among infected and eggnegative individuals, they were limited to the initial evaluation before treatment. Anti-SEA
and SWAP IgG4 levels were significantly higher in infected individuals before and after
treatment, when compared to egg-negative individuals. Significant association was observed
only to SEA-specific antibodies at time zero, showing a correlation between intensity of
infection and IgE (r = -0.266, P = 0.012), and also between geometric mean egg counts and
IgG4 (r = 0.239, P = 0.025). Similar association was observed between parasite burden, IgG4
to SWAP (r = 0.502, P = 0.015) and parasite burden and IgE to SEA (r = 0.470, P = 0.002),
one and three years after treatment, respectively. Conclusions: Our results contribute to
further evaluate of the use of specific IgE and IgG4 ELISA to predict susceptibility/resistance
to human schistosomiasis and suggest that the balance between the reactivity of these
two antibody to Schistosoma antigens determine the susceptibility to reinfection. Financial
support: NIH-ICIDR Grant 1R03AI071057-01, FAPEMIG, CNPq, CAPES.
311
SCHIStOSOMaL MYELORadICULOPatHY: a 10-YEaR fOLLOW-UP StUdY
Sílvio Roberto Sousa-Pereira; thiago Cardoso vale; Paloma fonseca; tânia antunes Carvalho;
Luciana Cristina dos Santos Silva; Izabela voieta; Carlos Maurício de figueiredo antunes; José
Roberto Lambertucci
Introduction: Schistosomal myeloradiculopathy (SMR) is the most severe and disabling ectopic
form of Schistosoma mansoni infection. In order to prevent severe and irreversible lesions,
early diagnosis and subsequent treatment of SMR are critical. Follow-up studies of patients
with SMR are lacking in the medical literature. Here we report a follow-up of patients with SMR
who regularly attended a Belo Horizonte University Hospital out-patient clinic for a time-period
of up to 10 years. Methodology: This is a prospective historical study of a population who live
in endemic areas for schistosomiasis in the Minas Gerais State, Brazil. From January 1998 to
January 2008, 38 out of 82 patients (46.3%) with myelopathy had a diagnosis of SMR confirmed
and were included in the present study. The following criteria were used for the diagnosis of
SMR: 1. clinical evidence of low spinal cord lesion; 2. demonstration of exposure to schistosomal
infection by microscopic or serological techniques and 3. exclusion of other causes of transverse
myelitis by magnetic resonance imaging and laboratory tests. Outcome was defined as “full
recovery” when the patient had complete improvement of neurological disease; as “partial
recovery without functional limitations” when the patient remained with only minor deficits
that did not interfere with daily activities; as “partial recovery with limitations” when the
patient was left with permanent disabling weakness (not able to ambulate without help), severe
urinary and/or intestinal dysfunction (retention or incontinence of feces or urine), or sensory
loss. Results: Twenty-six patients (68.4%) were male; the ages at diagnosis ranged from 11 to
59 years (mean: 28±12). All patients were treated and followed at the out-patient clinic every 6
months. Medium follow-up was of 31 months (ranging from 1 to 120 months). Sixteen patients
(42.1%) received prednisone (1mg/kg, body weight, single dose) whereas another 16 (42.1%)
received methylprednisolone (1g/day) for 5 days followed by oral prednisone (1 mg/kg, body
weight, single dose) for 6 months. Schistosomicidal drugs, such as praziquantel, oxamniquine or
both in combination were prescribed in 65.8, 7.9 and 5.3% of the patients, respectively. Twelve
patients (31.6%) experienced adverse reactions to steroids, such as weight gain, cushingoid
face, systemic arterial hypertension, acne, dyspepsia and bilateral avascular necrosis of femoral
heads. Twenty eight patients (75.7%) walked independently, 7 (18.9%) walked with support
and 2 (5.4%) were confined to a wheel-chair. Notwithstanding proper treatment, 19 (50%) of
the patients persisted with sphincter dysfunction (incontinence or urgency), 16 (42.1%) with
neuropathic pain, 12 (31.6%) with lumbar pain, 11 (28.9%) with intestinal impairment, 10 (25%)
with reduced patellar reflexes, 7 (18.4%) with erectile complaints, 6 (16.2%) with Babinski sign
and 5 (13.9%) with increased patellar reflexes. A partial recovery without functional limitations
outcome was observed in 22 patients (57.9%), followed by a full recovery outcome in 11 (28.9%)
and a partial recovery with limitations in 5 (13.2%). One patient (2.6%) presented with an
alternative diagnosis during the follow-up period. Conclusions: In short, our data demonstrate
that most patients responded well to treatment (partial recovery without functional limitations).
Full recovery was observed in almost a third of the patients and an error in diagnosis was
observed in only one patient. Side effects of the steroids were noted in a third of the patients.
312
SCHIStOSOMICIdaL aCtIvItY and tEgUMEntaL aLtERatIOnS IndUCEd BY PIPLaRtInE
On SCHIStOSOMULa and adULt fLUKES Of SCHISTOSOMA MAnSOnI
Josué de Moraes; Carlos nascimento; Eliana nakano; Lydia f. Yamaguchi; Massuo J. Kato;
toshie Kawano
USP, SP - Brasil
Introduction: Schistosomiasis is one of the most common human parasitic diseases in the
world. Treatment relies on a single drug (praziquantel) to eliminate the adult worms but,
this has no prophylactic properties and is ineffective against larval schistosomes. In addition,
given the potential for drug resistance, it is prudent to search for novel therapeutics. In this
context, the search for bioactive natural products against schistosome has great importance
for establishing future strategies to control schistosomiasis. Piplartine is an amide found in
several Piper species (Piperaceae). This amide has shown several biological activities such as
antifungal and insecticide as well as trypanocidal and antileishmanial effect. The aim of the
present study was to determine the effect of piplartine isolated from Piper tuberculatum
against schistosomula and adult flukes of Schistosoma mansoni. The efficacy of this amide
was examined regarding: a) schistosome survival; b) reproductive fitness; c) motor activity;
and d) alterations on S. mansoni tegumental surface examined through confocal microscopy.
Methodology: Cercariae are mechanically transformed into schistosomula and adult worms
were collected by perfusion method. Schistosomula and adult worm pairs (male and female
coupled) were each incubated in vitro using piplartine over a wide concentration range (1–
200 µg/ml). Different concentrations of praziquantel were used as positive control groups.
Worm motor activity, egg output (oviposition), tegumental alterations, and survival of the
parasites were monitored on daily basis for 5 days using an inverted microscope and a
stereomicroscope. Tegumental changes and the quantification of the number of tubercles
were performed using a confocal microscope. The cytotoxicity activity of the piplartine was
evaluated on monkey kidney fibroblasts (Vero cells) at different concentrations; the cytotoxicity
was determined by the crystal violet method. Results: The survival of schistosomula and
49-day-old adult worms of S. mansoni was assessed in vitro by incubation with different
concentrations of piplartine. This amide significantly reduced worm motor activity and caused
death of all larval- and adult-stages schistosomes within 24 h at 25 and 5 µg/ml, respectively.
Regarding the effects of piplartine on adult worms, no significant difference was observed
in the mortality rate between male and female parasites. Considering the strong lethal
effect of piplartine on adult schistosomes, the in vitro oviposition was continually monitored
to assess the sexual fitness of treated worms. At the highest sub-lethal concentration for
adult worm (2 µg/ml), an inhibition of 75% in egg laying was observed despite of parasites
remained coupled. In addition to the mortality rate and changes in the motor capacity of S.
mansoni adults, the results highlighted the effect of piplartine on the parasite’s tegument.
The morphological alterations of the tegument occurred in a dose-dependent manner and
were more pronounced in male than in female adults. The changes in the tegument surface
included peeling as well as formation and collapsing of tubercles. To further describe the
effects on the tegument, we performed the confocal microscopy analysis, which showed
alterations caused by piplartine on the tubercles in a dose-dependent manner at doses
higher than 5 µg/ml. For example, the number of intact tubercles in an area of 20,000 µm2
313
on the schistosome´s tegument in male worms of the negative control was 49, while in the
group exposed to 200 µg/ml of piplartine was 2. Additional morphological aberrations were
observed in adult worms after 2-24 h of incubation with 100 to 200 µg/ml of piplartine;
moreover, at this concentration range, all adult male schistosomes showed morphological
changes on the oral and ventral sucker regions. In contrast, female schistosomes showed
tegument damage caused by piplartine but only at doses higher than 50 µg/ml. Regarding
the schistosomula stage, piplartine caused alterations on the tegumental surface of parasite,
associated with a reduction in body lengths, in a dose-dependent manner in the range 25
to 100 µg/ml. To examine whether piplartine is tolerated by mammalian cells, we incubated
Vero cells with piplartine at different concentrations that were lethal to S. mansoni adult
worms (3 to 10 µg/ml). The cytotoxicity was not detected in Vero cells after the treatment
with piplartine at the maximum dose tested. The cells exposed to piplartine at 3, 4, 5, and 10
µg/ml during 2, 24, 48, 72, and 96 h, had no noticeable effects on their viability. Conclusions:
These findings revealed that piplartine is an effective compound against larval- and adultstages of S. mansoni in vitro. The mortality rate and the extensive morphological tegument
alterations for both schistosomula and adult stages of the parasite, combined with its effect
on reproductive fitness, it is clear that piplartine is a promising antihelminthic agent.
314
SCHIStOSOMICIdE EffECtS Of nEW tHIazOLIdInE dERIvatIvES (ExPERIMEntS In
vItRO)
anekécia Lauro da Silva; Sheilla andrade de Oliveira; andréia ferreira de Barros; veruska
Cíntia alexandrino de Souza; antônio Sérgio alves de almeida Júnior; Roni Evêncio de araujo;
Maria do Carmo alves de Lima; valéria Rêgo alves Pereira; Ivan da Rocha Pitta; Suely Lins
galdino
Introduction: Schistosomiasis is still considered to be a serious public health problem.
Treatment for this parasitic disease is based on chemotherapy, praziquantel being the
drug of choice for being more effective against all Schistosoma species. However, the
exclusive use of praziquantel in the treatment of intestinal schistosomiasis may account
for the development of resistance to the drug in some patients. The present study aims
evaluate in vitro the efficacy of some thiazolidine derivatives on Schistosoma mansoni adult
worms. Methodology: Male Swiss mice (Mus musculus) were submmited to percutaneous
infection with 120 recently shed Schistosoma mansoni cercariae obtained from laboratoryraised Biomphalaria glabrata snails (Belo Horizonte strain). After 60 days of infection,
worms were removed from the mesenteric and portal veins and kept in RPMI medium at
37ºC in a humid 5% CO2 atmosphere. For in vitro tests thiazolidine derivates LPSF/SF-22,
LPSF/SF-25 and controls with praziquantel were used in different concentrations (100μg/
mL, 80μg/mL, 60μg/mL e 40μg/mL). Results obtained are the average of duplicate samples
for each concentration. Parasites were kept for 6 days and watched every 24 h to evaluate
their general conditions (motor activity and mortality rate). The cytotoxicity of the above
compounds was determined using BALB/c mice splenocytes. Results: LPSF/SF-22 and LPSF/
SF-25 compounds showed activity in vitro, with a mortality rate of adult worms of 70% and
86% respectively, in the 80μg/mL concentration. In cytotoxicity tests, thiazolidine derivative
LPSF/SF-25 showed a better cytotoxicity than LPSF-22. Conclusions: Our results showed that
LPSF/SF-25 thiazolidine derivative can be a future candidate as schistosomicide. However,
studies must be performed in vivo for a better evaluation of therapeutic potential of these
compounds.
315
tHE IMPORtanCE Of EaRLY dIagnOSIS In nEUROSCHIStOSOMIaSIS: CaSE REPORtS
vinícius Martins alecrim; tatyana Layla aguiar Hazin; Joelton Barbosa da Silva; Marcos daniel
nigro da Silva; Rodrigo argenta toniolo; ana virgínia Matos Sá Barreto
Centro de Pesquisas Aggeu Magalhães, PE - Brasil
Introduction: Schistosomiasis is a parasitic disease caused by Schistosoma mansoni,
endemic in rural Pernambuco and being characteristic of poor rural areas with low socioeconomic development. The neuroschistosomiasis (myeloradiculopathy or spinal cord
schistosomiasis) is a complication of ectopic schistosomiasis and has being increasingly
reported in medical literature. Back pain or lower limb (LL), bladder dysfunction, weakness
of lower limbs, lower limb numbness and impotence are shown as clinical features.
Adult males with average age 26-29 years are most affected. Early diagnosis and prompt
institution of anti-inflammatory therapy are essential for determining the prognosis
of the disease and to prevent irreversible neurological damage. Treatment consists in
the association of antischistosomal drugs (praziquantel and Oxamniquine) and steroids
(prednisone). The present study deals the story of medical intervention in two patients
with signs and symptoms of neuroschistosomiasis. Methodology: Case Records: 1) JEN, 21
years old, male, born and raised in Bom Jardim, Pernambuco, Brazil. Patient was admitted
at the Hospital das Clinicas at Universidade Federal de Pernambuco (HC/UFPE) with
schistosomiasis diagnosis and weakness in lower limbs and pain in the lumbar region. The
examination of cerebrospinal fluid (CSF) was negative for helminth. The electrophysiological
study was compatible with myeloradiculopathy with moderate damage due to the second
lumbar vertebra to the second sacral vertebra. JEN was treated with the drug combination
(Praziquantel and Prednisone) and was referred to neurofuncionnal physical therapy. After
two years of treatment, in 05/10/2009, the patient was considered cured to form the
spinal cord schistosomiasis. 2) RMS, 23 years old, male, born and raised in Camaragibe,
Pernambuco, Brazil, was admitted at HC/UFPE. The initial symptoms were fever and back
pain. Examination of stools revealed the presence of S. mansoni. However, the CSF showed
negative results. An ultrasound at HC concluded that the patient presents hepatointestinal
form of schistosomiasis (HIE), and at this period the patient was in a wheelchair. There is
no report about the patient‘s treatment for neuroschistosomiasis using the association
of drugs (praziquantel and prednisone). RMS has evolved into a condition of permanent
sequelae of medullary. Results: Discussion: The neuroschistosomiasis (NE) is a severe
developing schistosomiasis infection, where the nervous system is compromised. It is
very important that an effective therapy is applied early to combat the possible sequels
disease in the future. The association of anti-inflammatory steroids (prednisone) and
drugs esquistomicidas (Oxamniquine or Praziquantel) is considered effective and may
contribute to a good prognosis. The patient number one was treated in early stages of
the disease with neurofuncionnal physical therapy and was considered cured. But, in
the patient number 2 was not adopted the same conduct, developing a condition of
permanent sequelae of medullary. It is important for the doctors of health care centers
from S. mansoni endemic areas remain alert to cases of neuroschistosomiasis. However,
the situation observed in these places does not allow, or difficult to identify early disease.
Hospitals and professionals responsible for health are not prepared for early diagnosis,
316
receive and treat patients with neuroschistosomiasis. In addition, the population of the
endemic areas also should be alerted of the ectopic schistosomiasis forms, its risks and the
importance of an early diagnosis and treatment. Conclusions: Conclusion: Early diagnosis
can result in an effective treatment against the neuroschistosomiasis, although the total
cure of patients with schistosomal myelopathy is not possible, it is important the early
medical intervention in order to minimize the damage caused by the establishment of
infection, as shown in this case records.
317
tHIazOLIdInE dERIvatIvES In tHE tREatMEnt Of MICE aCUtELY InfECtEd WItH
SCHISTOSOMA MAnSOnI
anekécia Lauro da Silva; Sheilla andrade de Oliveira; andréia ferreira de Barros; veruska
Cíntia alexandrino de Souza; Jamerson ferreira de Oliveira; Roni Evêncio de araujo; Maria do
Carmo alves de Lima; Ivan da Rocha Pitta; Suely Lins galdino
Introduction: Schistosomiasis is a chronic disease that affects over 200 million people
worldwide. For some species of Schistosoma infecting humans, praziquantel is the
drug of choice. However, the exclusive use of praziquantel (PZQ) in the treatment of
schistosomiasis may account for resistance of Schistosoma mansoni to the drug. Since
tests with the thiazolidine derivative LPSF/SF-25 showed in vitro a mortality rate of adult
Schistosoma mansoni worms, the therapeutic efficacy of the compound was studied in vivo.
Methodology: Swiss mice (weighting 20 g on average), infected with 120 S. mansoni cercariae
(Belo Horizonte strain), by subcutaneous route. Sixty days after exposure, the animals were
randomly divided into two experimental groups: Group I (LPSF-25/25mg/Kg); group II (LPSF25/200mg/Kg) and three control groups: Group III (PZQ/25mg/Kg); group IV (PZQ/200mg/
Kg) and group V (without compound). Two weeks after treatment, mice were euthanized
and perfused. The body cavity, liver, and mesenteric veins were examined for worms after
perfusion, to ensure all parasites were removed. The efficacy of treatment was measured as
the percentage of reduction of worm burden based on worm counting. For oogram studies,
two 1cm fragments of intestine (terminal ileum) were obtained. In each fragment, all the
different egg stages were evaluated. Results: The thiazolidine derivative LPSF/SF-25 tested in
vivo showed only a 28% reduction in the number of the adult Schistosoma mansoni worms
as compared to 100% in praziquantel groups and absence of some evolutionary stages in the
parasites’ eggs. Conclusions: The thiazolidine derivative LPSF/SF-25 did not show a better
efficacy than as praziquantel. Further studies about parasite fecundity and other aspects are
still needed.
318
tREatMEnt WItH HARPAGOPHYTuM PROCuMBEnS MOdULatES EOSInOPHILIa In
ExPERIMEntaL MOdEL Of SCHIStOSOMIaSIS
Sandra Regina Pereira de Oliveira; Joice Margareth de almeida Rodolpho; Ricardo de Oliveira
Correia; debora Meira neris; Laís Cristina de Souza; naiara naiana dejane; Paulo César vieira;
fernanda de freitas anibal
Universidade Federal de Saõ Carlos, SP - Brasil
Introduction: Schistosomiasis mansoni is a disease caused by the trematode Schistosoma
mansoni that causes considerable morbidity and mortality around the world. Characterized
by the presence of granulomas, immunopathology resulting from residual cellular
infiltrates and fibrosis, mainly in the liver. In the infected host, the disease includes the Th2
immune response with increased IL-5 mainly in response to egg antigens. Eosinophils are
important cells in granuloma formation; their recruitment to sites of infection by antigen
of the egg is mediated by a combination of interactions between adhesion molecules and
chemokines. Chemokines represent a large family of low molecular weight cytokines with
selective chemoattractant property. They participate in the recruitment of leukocytes to
the inflammatory site to bind to its receptors in these cells. The chemokines that bind to
receptors CCR2 and CCR3 as CCL2, CCL3, CCL7 and CCL11 are important in regulating Th2
interleukins. EO are specifically recruited to the site of egg deposition, the cellular infiltrate
favors hepatic granuloma, and approximately 50% of these cells EO. Methodology: We used
swiss mice, female, weighing 18-20g, these animals were divided into groups infected and
treated infected (II, III and IV). Groups II, III and IV received daily doses of crude extract
of Harpagophytum procumbens via gavage at concentrations of 15, 30 and 60 mg / kg /
animal, respectively. The animals infected / treated or not, received 50 cercariae / animal
subcutaneously. After 48 days of infection the animals were euthanized and the number of
EO in blood and LCP were determined. Results: Results: Our results showed that the number
of eosinophils in blood and LCP increased significantly in untreated infected animals, when
compared with control animals, under the same conditions. This increase was observed
throughout the analysis period, 48 days after infection. However, infected animals that
were treated with Harpagophytum procumbens showed reductions in the number of
eosinophils in these compartments, we observed significant reduction this cells when
compared with animals only infected and untreated. Conclusions: Thus, these data suggest
that Harpagophytum procumbens activity presents a modulator of the immune response by
interfering with the migration of eosinophils in ways yet unknown.
319
ULtRaSOnOgRaPHICaL aSPECtS Of PERIPORtaL fIBROSIS In adOLESCEntS
InfECtEd BY SCHISTOSOMA MAnSOnI In an EndEMIC aREa Of SCHIStOSOMIaSIS In
PERnaMBUCO, BRazIL
Luciana Carvalho zani; Otávio Sarmento Pieri; tereza Cristina favre; aline favre galvão; ana
Paula Brás Pereira; ana Lucia Coutinho domingues
Instituto de Pesquisa Evandro Chagas, Fiocruz, RJ - Brasil
Introduction: Schistosomiasis is one of the most prevalent neglected tropical diseases in
the Americas, with the number of carriers estimated at 7.1 million, 95% of which in Brazil.
Early diagnosis followed by prompt treatment of infected people can prevent severe
morbidity and contribute to its reversal. This work identifies the profile of morbidity in
adolescents (10-19 years) before and after of treatment with either 40mg/kg or 60 mg/kg
praziquantel (PZQ) through qualitative analysis of periportal fibrosis (PF) and its changes by
ultrasonography, linking it to the intensity of infection, PZQ dose and sex. Methodology: A
total of 196 adolescents with more than 100 eggs of Schistosoma mansoni per gram of faeces
(epg) by the Kato-Katz method (two exams, two slides each) were recruited for a doubleblind, randomized, clinical trial in the municipality of São Lourenço da Mata. A cohort of 167
individuals was followed at six and twelve months after treatment. The presence of PF and
image patterns (A through F) were designated according to the WHO protocol (Niamey).
Differences between the image pattern of PF and the intensity of infection (estimated by the
geometric mean of log 10 - transformed (x +1) before and after treatment were evaluated by
ANOVA. The association between dose of PZQ and the proportion of subjects who improved
the image pattern of FP at six and twelve months after treatment was evaluated by odds ratio
(OR) with 95% confidence interval (CI95), obtained through logistic regression. Results: In
patients treated with 40mg/kg of PZQ, the geometric mean was 313.1 epg before treatment,
0.96 epg at six months and 2.31 epg at twelve months. In those treated with 60mg/kg the
geometric means were 350.6 epg, 0.13 and 0.97 epg, respectively. Overall prevalence of PF
was 46.1% before treatment, 43.7% at six months and 34.7% at twelve months. The following
image patterns of PF were found before treatment: A (46.7%), B (23.9%), C (28.1%) and D
(1.2%). Six and twelve months after treatment, the patterns were, respectively: A (51.4%
and 60.4%), B (19.1% and 8.4%), C (26.9% and 29.3%) and D (2.4% and 1.8%). There was
no significant difference in infection intensity between the different image patterns of PF.
There was no significant improvement of image patterns of PF between the groups (40 and
60 mg/kg) either six or twelve months after treatment. The proportion of females (37.8%)
who improved the image pattern of PF at 12 months was significantly higher than that of
males (21.8%) (OR = 2.263; CI95: 4.491 - 1.140). There was no significant improvement in
the image pattern of PF in relation to other variables at six and twelve months. Conclusions:
The reduction in cases of recent PF (image pattern B), which occurred regardless of the PZQ
dose, confirms previous findings that treatment prevents severe cases (image pattern C
e D). The severity of PF was not related to the geometric mean epg, probably due to the
moderate-intensity infections that prevail among adolescents in that endemic area. It is
recommended that active case-finding followed by treating the infection carriers should be
strongly implemented at the primary health care level as a preventive measure for morbidity
control in the endemic areas.
320
URInaRY IMMUnOLOgICaL PROfILE Of HEPatOSPLEnIC SCHIStOSOMIaSIS PatIEntS
José Roberto Lambertucci; alba otoni; Izabela voieta; Carlos Maurício antunes; antônio Lúcio
teixeira
Introduction: The renal involvement on Schistosoma mansoni can complicate the
helminthiases’ evolution, raising substantially the disease’s social-economic cost. (WHO,
2008). The objective is identify urinary interleukins levels as mediators of renal injury in
hepatosplenic schistosomosis (EHE) patients. Methodology: It’s a pilot study in which were
selected: 18 patients with a mean of 45,5±11 years old with hepatosplenic schistosomosis
diagnosis; and 21 healthy volunteers (VS) with a mean of 40 years old. The patients were
submitted to clinical and laboratory analysis, excluding glomerulophaties secondary causes.
In both groups albuminuria was submitted to analysis in 24-hour urine sample, and the
following interleukins were measured in the urine: IL8, IP-10, MCP1, MIP1α, RANTES and
eotaxin-2. Results: Group 1 (VS) = absence of albuminuria (n=21). Group 2 = EHE patients with
<300mg/24 hours (n= 15) albuminuria and Group 3 = EHE patients with ≥300mg/24 hours
(n=3) albuminuria. The RANTES interleukins (p<0, 000) IL8 (p<0, 000) and MCP 1 (p<0,018)
presented significantly higher levels in Group 3 compared to Group 1 and Group 2. The
eotaxin-2 (p<0, 009) presented significant difference comparing all three groups, however its
values were lower in Group 3. IP10 and MIP1α revealed no statistical significance between
the three groups Conclusions: In EHE patients, who presented albuminuria ≥300mg/24 hours
(Group 3), the interleukins IL8, MCP 1 and RANTES levels were higher and the eotaxin-2
presented lower levels. These results suggest that this is a possible immunological profile of
renal injury mediator in EHE patients
321
vaRICEaL UPPER gaStROIntEStInaL BLEEdIng In EMERgEnCY HOSPItaL In RECIfE,
PERnaMBUCO
thais cavalcanti de almeida; ana Lúcia Coutinho domingues; José Roberto de almeida; admar
Borges da Costa Junior; adriana g. de Moura; Roberta Cavalcanti de almeida
Departamento de Medicina Clínica da UFPE, PE - Brasil
Introduction: ABSTRACT: Upper gastrointestinal bleeding (UGB) caused by esophagogastric
varices is the most serious complication of portal hypertension, with high rates of rebleeding
and mortality. In Pernambuco, where schistosomiasis mansoni is endemic, this type
of bleeding occurs frequently. The aim of this work is to observe, in patients who have
Upper gastrointestinal bleeding caused by esophagogastric varices assisted at the upper
digestive endoscopy unit of Restauração Hospital in Recife, which is the reference unit
for the attendance of digestive bleeding emergency, the hepatic illness that causes portal
hypertension through the hepatic aspect of the ultrasound, the rates of death and rebleeding
and check the orientations that the patients received after being discharged about the use
or no of beta-blockers. Also, labs and physical exams were done, medical histories were
checked. Methodology: Patients and methods: This is a descriptive case series study about
UGB patients assisted at the digestive endoscopy unit of Restauração Hospital in Recife from
October 2008 to October 2009. Data on medical history, clinical and demographical aspects
were collected and an endoscopy was performed to identify the area of the bleeding, as
well as a propaedeutical endoscopy to control the bleeding, when necessary. Afterwards,
when the patients were stable, an abdomen ultrasound was performed and labs were run to
diagnose the underlying disease. The patients received follow up over the phone for three
months after being discharged, in order to obtain the rebleeding and death rates. Results:
Results: Out of the samples of 385 UGB patients, 183 had UGB caused by esophagogastric
varices, 5 of which couldn’t be included in the study because of the gravity of their cases,
due to their impossibility of being submitted to exams after the endoscopy, so 178 patients
were analyzed. The specific-age rate was 53,9 years old, with 115 (64,6%) males. The most
frequent age group was from 50 to 59 years old (31,5%) and 67 (37,7%) were born from the
“Zona da Mata” region. Physical examination revealed ascitis in 58 (32,6%) patients. It was
seen that 177 patients had esophageal and 78 gastric varices. The blood tests done after
the endoscopy to identify the underlying disease showed approximately 2,8g/dl of albumin,
total bilirrubin of 2.1mg/dl, INR 1,4 and also that 12 patients had hepatitis B and 9 had
hepatitis C and 84 (47,2%) related alcohol abuse. Hepatic chronic disease (HCD) was defined
with the ultrasound in cirrhosis in 38 (31,7%) patients, schistosomiasis in 60 (50%) and mixed
disease in 22 (18,3%). After hospital discharge, only 57 out of 174 (32%) contacted patients
were granted specialized ambulatory care from the public system and 45 (25,3%) were in
use of propranolol. Three months after the UGB episode, rebleeding was found in 92 out of
161 (57,1%) contacted patients and there were 44 (27,3%) deaths, 16 out of 44 (36,4%) of
which were during hospital stay and 28 out of 44 (63,6%), after being discharged. There was
a correlation between the rebleeding and the deaths (p<0,001), between the rebleeding
and the presence of red spots on the esophageal varices (p<0,001), between the rebleeding
and the diagnose of cirrhosis and mixed hepatic chronic disease by ultrasound (p=0,023),
between the rebleeding and the presence of ascitis (p=0,004) and between the rebleeing
322
and the absence of propranolol use (p=0,002). There was also a relation between death and
age 50 to 59 years (p=0,016) and the hepatic aspect detected by ultrasound (p=0,001), as
well the presence of periportal fibrosis in the USG with the birthplace (p=0,031). Conclusions:
Conclusion: In this study, it was possible to observe that there were predominance of male,
bleeding from esophageal varices, patients from the high endemicity of schistosomiasis in
the state and were observed elevate rates of rebleeding and death. So, this work was done
to call the attention of the authorities to they promote improvement in the public health
services, which can lower both morbidity and mortality rates.
323
aCUtE HEPatOSPLEnOMEgaLY aLtERatIOnS In MICE InfECtEd BY SCHISTOSOMA
MAnSOnI and tREatEd WItH n-aCEtYLCYStEInE and/OR PRazIQUantEL
andré de Lima aires; tiago Moreira alves feitosa; Renata alexandre Ramos da Silva; Juliana
Kelle de andrade Lemoine neve; zilma Perreira dos anjos; giuliana viegas Schirato; Elizabeth
Malagueno de Santana; Mônica Camelo Pessoa de azevedo albuquerque
UFPE, PE - Brasil
Introduction: The main sites for tissue alterations by schistosomiasis mansoni is liver
and spleen due to the constant deposition of a great number of worm eggs causing the
hepatosplenomegaly, the most important and severe clinical finding of this disease resulting
in 500,000 deaths per year approximately. Therapy with praziquantel (PZQ) is efficient but it
does not change the inflammatory course already installed that may lead irreversible sequels.
N-acetylcysteine (NAC) has been employed in attempt to attenuate damage observed in
chronic liver disease because it presents anti-inflammatory properties as well as antioxidant
and hepatic detoxifying effects. The present study evaluated the NAC effects, individually
or associated to PZQ, on levels of hepatosplenomegaly in schistosomal mice. Methodology:
Forty 30-day-age Swiss Webster female mice percutaneously infected with 50 cercariae of
S. mansoni (BH strain) and 40 not infected mice were utilized in this experiment. These two
groups (infected and not infected) were subdivided in 4 subgroups with 10 animals according
to the following therapeutic scheme: NAC, PZQ, NAC + PZQ, and control group that received
only the drug vehicle. N-acetylcysteine was orally administered, at 200 mg/Kg/day, during
60 consecutive days after infection. Treatment with PZQ occurred during five consecutive
days from 45th day at 100 mg/Kg/day, orally. Every seven days after infection the animals
were weighed to determine weight curve for subsequent relationship with the weight of
liver and spleen. At the end of treatment, animals were sacrificed to remove liver and spleen
for subsequent weighing. Results: With relation to body weight, there were not significant
differences (p > 0.05) in the intergroup and intragroup analyses during all experiments. At
the end of therapy, means of body weight of not-infected animals and infected animals
were 41.87 g (± 1.45) and 39.18 g (± 1.0) respectively. Independently of therapeutic scheme
adopted in this study, all subgroups of infected mice presented an increasing in the weight of
spleen and liver as compared to not-infected animals. Thus, this fact evidences the functional
alteration of these organs caused by schistosomiasis. Animals of not-infected subgroups
treated with NAC and/or PZQ did not exhibit significant difference (p > 0.05) on spleen and
liver weights. However, those mice infected and treated with PZQ and NAC + PZQ showed
important percentage reduction in the weight of spleen of 40.24% and 50.30% respectively
when compared to infected controls. For the liver weight, only association NAC + PZQ
exhibited significant percentage of remission of 24.18% as compared to infected animals of
control groups. Conclusions: From our results and considering the adopted therapy in this
study, we could conclude that administration of NAC and/or PZQ does not change the body
weight as well as the spleen and liver weights in healthy mice. However, in schistosomal
mice, hepatosplenomegaly levels were reduced after treatment with NAC and/or PZQ.
324
aCUtE nEUROSCHIStOSOMIaSIS: a REPORt On tHREE CaSES and REvIEW Of tHE
LItERatURE
thiago Cardoso vale; Sílvio Roberto de Sousa-Pereira; Izabela voieta; José Roberto Lambertucci
Introduction: Acute schistosomiasis may be complicated by life-threatening neurological
involvement. The acute phase occurs 2 to 6 weeks after bathing in infected waters.
Neurological symptoms over the course of acute schistosomiasis are frequently accompanied
by fever and eosinophilia. Patients may become confused and develop focal motor deficits or
seizures when the brain is impaired. Acute schistosomal myeloradiculopathy can also occur,
manifested by acute lumbar pain followed by lower limb weakness and sphincter disturbances.
Methodology: We describe three cases of acute schistosomiasis complicated by neurological
involvement. Patient ages were 27, 22 and 23 years old. Schistosomiasis diagnoses were
based on epidemiological, clinical, imaging and brain biopsy data. All subjects were submitted
to brain or spinal cord magnetic resonance imaging before and after treatment. All were
treated with a schistomicidal drug (praziquantel) and steroids. Results: Case 1: a 27-yearold woman who presented with a 2-week history of melena, weight loss, fever, lower-limb
pain and paresthesia, followed by urinary and fecal retention. She was previously exposed
to water populated by cercariae-infected snails. Neurological examination revealed inferior
motor neuron syndrome affecting her lower limbs. A positive ELISA IgM anti-surface egg
antigen and a rectal biopsy containing granulomas with Schistosoma mansoni eggs confirmed
the exposure. Lumbar resonance imaging showed a lesion at the distal portion of the
conus medullaris, extending towards L1 and L2 spinal cord levels, and with heterogeneous
contrast enhancement. Cerebrospinal fluid revealed pleocytosis with an increase number of
mononuclear cells and proteins (1440 mg/dl). Praziquantel and steroids were started after 2
months of symptom onset and the patient presented with significant improvement of pain
and paresthesia, but urinary and fecal retention persisted for 5 months with the need of
intermittent catheterization. In her latest follow-up consultation, she complained of discrete
sphincter disturbance without the need of urinary catheterization. Case 2: a 22-year-old
man who was admitted to hospital with a 2-day-history of fever, diarrhea, low back pain and
asymmetrical lower limb pain and weakness, particularly on the left side, followed by urinary
retention. A history of bathing in water populated by cercariae-infected snails was present 1
month earlier. Neurological examination revealed weakness on his left triceps surae muscle
and on both iliopsoas. Left Aquile’s tendon reflex was absent. Cerebrospinal fluid examination
revealed lymphocytic pleocytosis with elevated proteins (75 mg/dl) and eosinophils (5 cells).
Serum eosinophilia was present (924 mg/dl). Parasitological stool examination revealed
the presence of S. mansoni eggs. Lumbar resonance imaging showed enlargement of
conus medullaris and signal abnormalities with contrast enhancement extending towards
T10-T12 spinal cord levels. Praziquantel and metilprednisone were prescribed immediately
after hospitalization. Oral steroids were taken for 6 months and the patient improved
significantly, though remained with a left distal lower-limb paresis in a 2 month follow-up.
Case 3: a 23-year-old female who presented seizures and headache 2 months after bathing
in an endemic area for schistosomiasis in Brazil. Brain magnetic resonance imaging showed
a contrast-enhanced left fronto-parietal nodular lesion which was biopsied revealing a well
325
formed granuloma around a S. mansoni egg. All patients are regularly seen in our institution
since the beginning of their illness. They are asymptomatic, functionally independent and
with no or minimal sphincter disturbance. Conclusions: Herein we describe 3 cases of acute
schistosomiasis with neurological complications. Two of them presented with myeloradicular
involvement and one with a cerebral tumoral lesion. All were treated with praziquantel and
steroids and partially recovered in a short period of time.
326
EffECtS Of SILYMaRIn In dMSO On ExPERIMEntaL SCHIStOSOMIaSIS MURInE
fabiana gonçalves Lino; Hílton antônio Mata dos Santos; Carolina Carneiro Rocha; fabíola
Ramos xavier; Letícia Campos da Costa; alexandre dos Santos Pyrrho
Universidade Federal do Rio De Janeiro, RJ - Brasil
Introduction: Schitosomiasis is parasite infection with an important public health impact. The
etiological agent is a Trematoda worm from genus Schistosoma. In Brazil only S. mansoni is
responsible for the infection and morbidity that affects people who lives in endemic areas.
Although, praziquantel is the drug to treat this disease, the sequelae from this infection could
be persevered for long time. Therefore, it is important identify a drug that could reverse or
attenuate the sequelae, mainly the hepatic fibrosis which leads to portal hypertension. Since
silymarin is reported by its hepatoprotector, hepatoregenerator, antifibrotic, and antineoplasic
action, it was evaluated this phytotherapic in murine schistosomiasis. Methodology: BALB/c
mice (6-7 weeks) were infected with BH strain cercariae. Silymarin were prepared in DMSO
5% or DMSO 1%. Mice were separated in non-infected (N), non-infected plus vehicle (N+
DMSO 5 %), infected (I), infected plus vehicle (I+ DMSO 5%), infected treated with silymarin
in DMSO 5% (I+ Sil 10mg/kgD5%) and silymarin in DMSO 1% (I+ Sil 10mg/kg D1%) group.
Animals were inoculated by intraperitonial route at intervals of 48 hours. At 55th day after
infection, the animals were submitted to euthanasia. Hepatosplenomegaly was evaluated
by the relation of organ and animal weight. The eggs distribution in hepatic, spleen and
intestinal was estabilished to evaluate if treatment could change the oviposition capacity
or worm maturation. The amount of hydroxyproline was measured in the hepatic tissue to
observe if treatment with silymarin could interfere with fibrotic process. It was also evaluated
the periovular granuloma area to verify if silymarin could interfere with this inflammatory
/ immunological aspects. Results: There were a reduction in hepatic and spleen index at
I+ Sil10mg/kg D1% in comparison with infected group (I). However, this reduction was not
observed in I+ Sil 10mg/kg D5% group. Moreover, there were differences between the I and
I+ Sil 10 mg/kg D5% as well I and I+ Sil 10 mg/kg D1% on reduction of periovular granuloma
area. In fibrosis evaluation, a reduction of hydroxyproline was observed in only both groups
treated with silymarin. The treatment did not interfere in oviposition capacity or in worm
maturation. Conclusions: The treatment with silymarin partially soluble in DMSO at low
concentration (1%) leads a reduction on hepatosplenomegaly and hepatic fibrosis, although,
silymarin in DMSO 5% reduced only the hepatic fibrosis, but not hepatosplenomegaly. In
both treatments with silymarin it was observed a reduction in periovular granuloma area.
Therefore, silymarin could be a promising complementary treatment to reverte/minimize
sequelae of this infection.
327
EvaLUatIOn Of tHE IntEStInaL MICROBIOta In MICE CHROnICaLLY InfECtEd BY
SCHISTOSOMA MAnSOnI
andré de Lima aires1,2; Kedma de Magalhães Lima1,2; thays Miranda almeida¹; tiago Moreira
alves feitosa2; Renata alexandre Ramos Silva²; Maria Helena Madruga Lima Ribeiro²; Mônica
Camelo Pessoa de azevedo albuquerque2; Célia Maria Machado Barbosa de Castro
Introduction: The infection by Schistosoma mansoni is characterized by the presence of
worm eggs which are the main pathogenic elements and the organs primarily affected
are liver, spleen and intestine. In this last organ, eggs that were retained on the intestinal
epithelium stimulate a granulomatous chronic inflammatory reaction resulting in
immunity disturbance. Gastrointestinal tract contain a complex and dynamic population
of microorganisms mainly composed of the bacteria composing the normal microbiota.
Investigations about this subject have attracted very interest in last years due to the
important role of microbiota in development and maintenance of the intestinal immune
response. It has been supposed that immunodepression, microbiota alterations and
failure of the defensive barriers of the intestinal mucosa are crucial factors related to
the passage of microbes and endotoxins from the intestinal lumen to the bloodstream.
Moreover, these factors are clinically observed in schistosomal individuals.This work
aimed to evaluate the intestinal bacterial microbiota of mice infected by Schistosoma
mansoni. Methodology: Twenty 30-day-age Swiss Webster male mice were divided in
two groups: not-infected (n = 10) and infected (n = 10) with 50 cercariae of S. mansoni
(SLM strain) percutaneously. After fifty days post-infection, it were realized individual
parasitological tests by mean of Kato-Katz method to confirm the infection of animals
and to estimate the parasite load related to the amount of eggs per gram of stool.
After 13th week of the cercariae contact, animals were sacrificed by mean of cervical
displacement. For the evaluation of the intestinal microbiota, stool of mice were directly
collected on sterile assay tubes from the median region of the small intestine. Collected
samples were immediately submitted to microbiological analyses. For qualitative
microbiological analysis, it was used the method recommended by Bisso (2008). In the
tubes containing stool samples, it was added sterile saline solution (NaCl, 0.9%) and
after mechanical homogenization it was performed five serial dilutions multiple of ten.
Of each dilution it was seeded 0.1 ml in chromogenic medium (Agar Biochrome, BIOLOG)
following incubation at 37ºC for 48 hours. Bacteria found on chromogenic medium were
biochemically identified and the results were expressed as colony forming unities per
gram of stool (CFU/g). Enterococcus, yeasts, Staphylococcus and coliforms which grew
in medium were also identified by their colonial characteristics and biochemical tests.
Results: All S. mansoni infected animals exhibited uniformity in the number of eggs
eliminated per gram of stool (60 ± 5.36). Generally, infected animals presented higher
variety and amount of bacterial colonies than the not-infected mice. Additionally, there
was an increasing of UFCs of mesophills, mainly coagulase negative Staphylococcus, and
coliforms, particularly Escherichia coli, in the infected group. There was no difference
(p > 0.05) with relation to the number of Enterococcus faecalis in both studied animal
groups. Conclusions: According to the results, the adopted experimental model
328
demonstrated that the relative immunodeficiency on animals infected by S. mansoni
influenced the imbalance in intestinal microbiota that may result in the translocation
of microorganisms and to associate to candidemia, bacteremia and multiple organs and
systems failure syndrome.
329
HEPatOSPLEnIC SCHIStOSOMIaSIS: CLInICaL and LaBORatORY CHaRaCtERIStICS Of
PatIEntS WItH and WItHOUt PULMOnaRY HYPERtEnSIOn
Claudio Lemos Morais; Juliana Papatella; Pedro Henrique Lima Prata; Izabela voieta; Carlos
Maurício figueiredo antunes; José Roberto Lambertucci
Universidade Federal de Minas Gerias, MG - Brasil
Introduction: Pulmonary hypertension has been described in 15% of patients with
hepatosplenic schistosomiasis in Brazil. Because no treatment for this complication was
available the disease received poor attention over the years. More recently, though, with
the advent of new vasodilators for treating pulmonary hypertension the natural history
of this complication is changing. A renewed interest is observed among researchers
and physicians in diagnosing and caring for this group of patients. Physicians should be
aware of the importance of pulmonary hypertension and also of new treatments. For
diagnosing this complication, clinical markers should be identified and validated. Here
we describe simple clinical findings and laboratory tests in hepatosplenic schistosomiasis
that should alert physicians for a presumptive diagnosis of lung disease and trigger a more
aggressive investigation of pulmonary hypertension. Methodology: Twenty patients (10
men) referred to the Tropical Diseases Outpatient Clinic with diagnosis of hepatosplenic
schistosomiasis, portal hypertension and indication for surgical intervention (splenectomy,
portal-variceal disconnection and suturing of esophagogastric varices) were selected for
this study. Schistosomiasis was defined by microscopic evidence of active infection (positive
parasitological stool examination or eggs in rectal biopsy) and a history of contact with
stream waters of an endemic area. Patients included in the study did not have any other
cause of chronic liver disease, such as cirrhosis, congestive heart failure or toxic or viral
hepatitis. Ages ranged from 34-65 years (47±8.4). Outpatient assessment included clinical
examination, abdominal US, serum markers of autoimmune hepatitis, hepatitis B and C
serology, blood cell count and liver function evalua¬tion (serum albumin and prothrombin
time). Clinical examination was done by one of us (CLM) with particular attention to
abdominal palpation and pulmonary auscultation. The right hepatic lobe was examined on
the anterior axillary line and the left lobe on a line passing through the xiphoid appendix.
The spleen was palpated and measured under the costal margin with the patient in dorsal
decubitus during deep breath. A pulmonary physical examination was performed to look
for typical signs of pulmonary hypertension; these include altered heart sounds, such
as a widely split second heart sound and a loud P2 or pulmonic valve closure sound. US
examination of the abdomen was performed using a real-time ALOKA SSD device 1700
(Japan) with electronic convex 3.5 MHz transducer, according to the WHO proto¬col for US
assessment of schistosomiasis-related morbidity (Niamey Working Group 1996). US hepatic
fibrosis was classified as absent (pattern A), slight (B, C, D and Dc), moderate (E and Ec) or
intense (F). The presence of collat¬eral vessels and the spleen length were evaluated. MR
was obtained using a GE 1.5 T Sigma unit (Milwaukee, USA). Axial and coronal 7-mm-thick
slices were performed in T1 and T2-weighted sequences, before and after gadopentetate
dimeglumine (Gd-DTPA) administration. MR analysis was guided by an adaptation of the
WHO protocol for US. All patients had a chest x-ray and computerized tomography of the
lungs. Those complaining of dyspneia underwent a comprehensive Doppler echocardiogram
330
with color flow mapping. Results: Seven out of 20 patients (35%), complaining of dyspnea
had pulmonary hypertension documented by Doppler echocardiogram and confirmed by
right-sided cardiac catheterization. In this group, systolic pulmonary pressure varied from
55 to 130mmHg. For the present analysis patients were divided into 2 groups: Group 1
comprised 7 patients with confirmed pulmonary hypertension and Group 2, 13 patients
without pulmonary hypertension. The mean platelet counts were 124,143mm³ (±69,996)
for group 1 and 58,667mm³ (±49,976) for group 2 (p=0.03). The spleen size was significantly
smaller in patients with pulmonary hypertension (Boyd 0-I: 71%; II-IV: 29% for group 1, and
Boyd 0-I: 12%, II-IV: 88% for group 2; p=0.002). Digestive bleeding was more common in
patients without pulmonary hypertension (14% in Group 1 versus 64% in Group 2; p=0.056).
Conclusions: In the present investigation, simple clinical observations (dyspnea, spleen size,
digestive bleeding) and laboratory data (platelet count) has been shown to be good indicators
of pulmonary hypertension. The findings of smaller spleens and higher platelet counts in
patients with pulmonary hypertension may be explained by the presence of large portoalpulmonary shunts. Patients with hepatosplenic schistosomiasis mansoni who present
those signs and symptoms should be further investigated to confirm or exclude pulmonary
hypertension, a severe, and lately, treatable complication of schistosomiasis.
331
HIStOPatHOLOgICaL and QUantItatIvE fEatURES Of SCHIStOSOMaL HEPatIC
gRanULOMaS In a MOUSE-MOdEL Of MEtaBOLIC PROgRaMMIng.
Christiane Leal Corrêa; Patrícia Cristina Lisboa; José Roberto Machado e Silva; Elaine de
Oliveira; Egberto gaspar de Moura; Renata Heisler neves; Regina Maria figueiredo de
Oliveira; adriana Cardoso gomes
Universidade do Estado do Rio de Janeiro, RJ - Brasil
Introduction: Poor quality of nutrition during early development is associated with adverse
health outcomes in adult life, a process referred to as developmental programming. In the
present study, a mice model of programming has been used for investigation of effects on
the schistosomiasis outcome in the offspring. Methodology: Three-month-old, virgin, Swiss
Webster female mice were housed together with a male mouse to mate. Lactating mice were
submitted to protein-restricted (PR, 8% protein) or energy-restricted (CR, calculation according
to the intake of the group PR) diet throughout lactation, whereas other pups received a
normal diet (C, 23% protein). From weaning on, all offspring mice were fed a normal diet until
120 days of age. Body weight, food consumption and central adiposity were monitored for
nutritional evaluation. For each dietary group, 10 mice, aged 2 months, were each infected
percutaneous with 50 Schistosoma mansoni cercariae (BH strain). Ten mice fed one kind of
diet group were kept uninfected as control. Infected mice were euthanized 9 wpi by cervical
dislocation for liver excision. Liver was processed routinely for histological preparation,
embedded in paraffin, sectioned at 5 µm thickness, and stained with hematoxylin and eosin,
Lennert’s Giemsa and Picrosirius for collagen plus polarization microscopy. Liver periovular
reactions were classified in: pre-granulomatous stages: weakly reactive or non-reactive and
exudative stages; granulomatous stages: exudative-productive, productive and involutional
granulomas. The area, perimeter, major, and minor diameter of individual granulomas were
measured by digital image analysis (Image Pro-Plus Media Cybernetics, US). All results have
been presented as mean values ±SE. One-way analysis of variance (ANOVA) and post hoc
Tuckey were used. Values of p < 0.05 were taken to be significant. Results: Among mice fed
restricted diet, PR mice showed lower food intake (6.013 ± 0.39 gram) compared to CR (10.89
± 0.30 gram; -38%) and C (9.30 ± 0.32 gram; -33%). This group showed lower weight (27.30
± 1.12 grams) than for C group (31.80 ± 1.29 grams; -14%; P < 0.05) and for CR mice (38.80
± 0.55 grams; -23%; P = 0.0001). CR mice had body mass heavier (38.80 ± 0.55 gram) than
control (31.80 ± 1.29 gram; +15%; P < 0.05). RP offspring showed higher liver granuloma
counting than other groups (+233% vs C; +200% vs RC; p<0.001). The qualitative cellular
composition of hepatic granulomas was similar among diet groups, with accumulation of
inflammatory cells (macrophages, eosinophils and lymphocytes), which behave differently
according to evolutive type of granulomas. During the exudative granulomas type, there
was a cellular richness of macrophages, eosinophils, neutrophils and lymphocytes. This
morphological picture, however, was slightly changed in exudative-productive granulomas
type, which was predominantly composed by fibroblasts and macrophages, whereas
lymphocytes, plasma cells and eosinophils showed less intensity. This group showed
higher number of exudative granuloma (+405% vs C; +2.930% vs RC, p<0.001) and lower of
exudative-productive granulomas (-44% vs RC, p<0.001). Morphometric analysis showed that
mean measurements of exudative granulomas from RC mice were smaller (5.94 µ2) than C
332
group (42.25 µ2) and PR group (76.97 µ2). However, exudative-productive granulomas were
smaller in C animals (59.50 µ2) than CR (96.76 µ2) and PR (108.02 µ2). Collagen content in
exudative granulomas showed higher values for CR mice (71 % vs C, p<0.05) and PR mice
(36 % vs C, p<0.05). Histopathological examination showed a normal portal space and liver
parenchyma (hepatocytes and sinusoids arrangement) which was irrespective of diets
assayed. In mice fed control diet, eggs elicited mono-and polymorphonuclear leukocytes
migration within the intravascular space. Also, fibrotic changes within the granulomas were
highlighted by the Masson’s trichrome stain. Photomicrographs of liver sections showed
exudative-exudative granulomas with a central egg surrounded by a prominent disarranged
cellular composition in the outer layer. CR mice showed a leukocyte infiltrate eventually
not-associated with the presence of schistosomal granuloma. Schistosome eggs elicited an
intense acute inflammatory reaction characterizing a pre-granulomatous type granuloma
and exudative peri-ovular lesions surrounding a central schistosomal egg. At cellular level,
mice presented pyknosis, karyorrhexis and karyolysis. Feeding of low-protein diet led to
microvesicular steatosis in the liver parenchyma with hepatocytes to show microvesicular
steatosis. Intense leucocyte infiltrate with collagen neogenesis and exudative-productive
granuloma with regular external contour were highlighted by the Masson’s trichrome
stain. Schistosomal granuloma showing concentric arrangement of reticular fibers and
liver regeneration characterized by binucleate hepatocytes were evidenced. Conclusions:
Nutritional changes associated with lactation may be an important factor influencing the
outcome of schistosomiasis, in which CR group seems to have evolutionary advantage in this
host–parasite relationship.
333
LIvER HIStOPatHOLOgY Of CHROnIC SCHIStOSOMIaSIS In MICE fEd HIgH-fat dIEt
alba cristina Miranda de Barros alencar; Renata Heisler neves; Marcele nogueira de Sousa
trotte; albanita viana de Oliveira; delir Corrêa gomes; José Roberto Machado-Silva
Universidade do Estado do Rio de Janeirto, RJ - Brasil
Introduction: High-fat diet feeding mice is accompanied by liver histopathological changes
such as prominent sinusoidal dilatations, microvesicular steatosis, increasing in connective
tissue, highly dilated smooth endoplasmic reticulum and necrosis in the hepatocytes.
Much of the morbidity of hepatic schistosomiasis mansoni has been learnt from animal
studies. Experimental studies show that high-fat chow alters the rate of both acute and
chronic schistosomiasis infection in mice. However, liver disorders are not known in chronic
schistosomiasis in mice fed high-fat chow. Objective: To investigate this gap based on light
histopathological investigation. Methodology: Female Swiss Webster mice (3 wk old) were
fed a high-fat chow or a standard chow. After feeding for a 6-month period and during
necropsy, total serum cholesterol (TC) was assayed, using the colorimetric enzymatic
method. Later, mice were infected by subcutaneous route with ~ 50 Schistosoma mansoni
(BH strain) cercariae. Uninfected mice were controls. Mice were divided into four groups:
uninfected mice fed standard chow (SC), uninfected mice fed high-fat chow (HFC), infected
mice fed standard chow (ISC) and infected mice fed high-fat chow (IHFC). At week 17 posinfection, mice were euthanized and necropsied for liver remove from each animal. The
liver was fixed, embedded in paraffin, sectioned at 5 µm and stained with hematoxilin and
eosin. Results: Among uninfected mice, TC levels before infection were: 62±13 mg/dL (HFC)
and 40±9 mg/dL (SC) (p= 0.0080) and 47±6 mg/dL (HFC), 50±12 mg/dL (IHFC), 30±10 mg/
dL (SC) and 35±7 mg/dL (ISC) (p= 0.0055), during necropsy. SC group showed normal liver
feature. The high-fat diet group (HFC) presented steatosis, discrete multifocal and periportal
mononuclear cell infiltrations and discrete proliferation of Kupffer cells. However, when
the high fat diet was combined with infection (IHFC group), histopathological examination
revealed steatosis, periovular granuloma, intense multifocal fibrosis and periportal
mononuclear cell (lymphocytes) and polymorphonuclear (eosinophil) infiltrations. The
ISC group showed vacuolar degeneration of the hepatocytes, periovular granuloma,
multifocal fibrosis, moderate multifocal and periportal mononuclear cell (lymphocytes) and
polymorphonuclear (eosinophil) infiltrations, and moderate proliferation of Kupffer cells.
Conclusions: The association of diet with infection seems to intensify liver injury during
chronic schistosomiasis.
334
PROdUCtIOn Of MaBS (MOnOCLOnaL antIBOdIES) antI-CO-StIMULatORY
MOLECULES fOR tHE tREatMEnt Of ExPERIMEntaL SCHIStOSOMIaSIS ManSOnI
Laís Cristina de Souza; Isabela gobbo ferreira; Ciro novaes Lino; denise Sayuri Calheiros da
Silveira; Bruna gabriela Silva; andré Barros; naiara naiana dejani; Joice Margareth de almeida
Rodolpho; Sandra Regina Pereira de Oliveira; débora Meira neris; Ricardo Oliveira teixeira;
Heloísa Sobreiro Selistre de araújo;Claudio alberto torres Suazo;auro nomizo;fernanda de
freitas anibal
Introduction: Schistosomiasis mansoni represents a major public health problem that
chronically infects more than 200 million people in developing countries. In the infected host,
the disease is characterized by the presence of granuloma, resulting immunopathological
cellular infiltration and tissue fibrosis. The disease caused in the host infected with S.
mansoni is mediated immune response, mainly by T cells, through the dissemination of
eggs that are in the liver and intestine, contributing to the development of granuloma. The
granulomatous process in schistosomiasis is dependent on CD4 + and requires recruitment
and accumulation of inflammatory cells at the site of deposition of eggs. Manipulations to
modulate the interaction between B7 molecules on antigen presenting cells (APCs) and costimulatory molecules (receptors) CD28/CTLA4 T cells may favor, in some circumstances,
blocking the immunological response in vivo. These co-stimulatory signals may have
different roles in various types of immune response. Thus, the aim of this study was to
produce MAbs anti the co-stimulatory molecules of T lymphocytes to subsequent treatment
of animals in the experimental model of schistosomiasis. Methodology: The MAbs were
obtained of cultivation of hybridoma 9H10 (anti-CD152) and PV-1 (anti-CD28) separately,
in bioreactor (spinner). The cultivation was done with 500 mL of RPMI 1640 containing
7.5% fetal bovine serum (SBF) and inoculum 106cel/mL, adequately oxygenated through
a tubular silicone membrane within which circulates pressurized air. Oxygen from the air
having a high permeability easily passes through the wall of silicone and then comes to the
culture medium where, after being dissolved, is consumed by the cells. During the culture
were collected aliquots at 12h intervals to assess the viability and expansion of hybridomas.
Results: After this process, we evaluated the development of hybridomas and quantified the
MAbs by ELISA in aliquots withdrawn at different times of cultivation. The 9H10 hybridoma
showed less viability and growth than the PV1. However, the production of MAbs was higher
than 9H10 in PV1. Conclusions: These data have to be better investigated. Thus, we suggest
different procedures for the cultivation and preparation of the samples during freezing and
thawing might have favored the deletion of one gene essential for the production of MAbs in
these cells PV1. Support: FAPESP
335
PULMOnaRY HIStOPatHOLOgY Of MICE
SCHIStOSOMIaSIS ManSOnI InfECtIOn
fEd
HIgH-fat
dIEt
On
aCUtE
vanessa Coelho de góes; Renata Heisler neves; alba Cristina Miranda de Barros alencar;
Marcele nogueira de Sousa trotte; albanita viana de Oliveira; delir Corrêa gomes; José
Roberto Machado-Silva
Universidade do Estado do Rio de Janeiro, RJ - Brasil
Introduction: Hypercholesterolemia is caused by multiple environmental factors and, plays
an important role in the development and pathogenesis of various human morbidities.
Pulmonary schistosomiasis, which is the most frequent complication outside guts on
Schistosoma mansoni infection, is expressed by pulmonary hypertension associated with
various diffused granuloma on tissues, mainly on the chronic phase. Previous experiments
from our laboratories showed that the schistosomiasis outcome was modified in mice fed
high-fat diet. However, lung damages in infected mice fed high-fat chow in acute phase has
not been investigated. OBJECTIVE: To investigate if the lung tissue are damaged in acute
schistomiasis infection in mice with underlying dyslipidemia, using light histopathological
investigation. Methodology: Twenty female Swiss Webster mice fed for six months a highfat chow (47% carbohydrates, 24% proteins, 29% lipids) were compared to control mice fed
a standard commercial chow, Nuvilab (74% carbohydrates, 22% proteins, 4% lipids). Blood
samples were collected for total serum cholesterol (TC) determination one day before
infection and nine weeks later. TC was evaluated by cholesterol esterase/cholesterol oxidase/
peroxidase method. Body mass was measured twice a week throughout the experiment.
After 6 months fed on diets, mice were transcutaneous infected with ~ 50 cercariae (BH
strain). Animals were divided in four groups: standard chow (SC), high-fat chow (HFC), infected
standard chow (ISC) and infected high-fat chow (IHFC). After 9 weeks post-infection (acute
phase) the animals were euthanized to remove the lungs. The organs were fixed, embedded
in paraffin, sectioned at 5 μm thickness and were stained with hematoxilin and eosin. Results:
Mice fed high-fat diet showed TC levels higher than animals fed standard diet (p<0.05).
However, TC serum levels were reduced in infected mice groups in both diets. All animals
from SC and HFC groups presented normal lungs. The ISC group presented discrete focal
mononuclear cell (predominancy of lymphocytes) and polymorphonuclear (predominancy
of neutrophil) alveolar infiltration in 10% mice. In other 10%, was observed a concentric
vessel endothelium hypertrophy, known as vasculitis, caused by hypersensitiveness to
schistosomula passage. On the IHFC group, alterations were observed in almost 50% of
mice. They presented vasculitis, discrete multifocal mononuclear cell (lymphocytes and
alveolar macrophage) and polymorphonuclear (predominancy of neutrophil) alveolar
infiltration and another focal mononuclear cell (lymphocytes and alveolar macrophage)
and polymorphonuclear (predominancy of eosinophil) periovular infiltration, suggesting a
granulomatous reaction. Conclusions: Schistosoma mansoni infection causes damages to the
lung tissue, especially when it is associated with high-fat diet.
336
SCHISTOSOMA antIgEnS dOWn MOdULatE tHE InfLaMMatORY IMMUnE RESPOnSE
In vItRO In PatIEntS InfECtEd WItH HtLv-1
Luciane Mota Lima; Silvane Maria Braga Santos; Ricardo Riccio Oliveira; Luciana Santos
Cardoso; Sergio Costa Oliveira; alfredo Miranda goes; alex Loukas; Edgar M. de Carvalho;
Maria Ilma araújo
Serviço de Imunologia, UFBA, BA - Brasil
Introduction: The human T cell lymphotropic virus type 1 (HTLV-1) is the causal agent
of Adult T cell Leukemia/lymphoma (ATLL) and HTLV-1-associated Myelopathy/Tropical
Spastic Paraparesis (HAM/TSP). HTLV-1 infects 20 million individuals worldwide.
HAM/TSP is clinically characterized by an insidious onset of spastic paraparesis, that
progressively leads to wheel chair dependency. Other manifestations associated with
HTLV-1 infection include Sjogren Syndrome, Polyarthritis, periodontal diseases, urologic
manifestations, Infective Dermatitis and Erectile Dysfunction. However, it is not known if
these clinical manifestations are associated with viral and immunologic factors or if they
represent risk factors for severe disease. HTLV-1 infects predominantly T cells and the
tax gene of the virus activates cytokine genes within T cells leading to the production
of large amounts of IL-2, TNF-α, IFN-ɣ and IL-15. Patients with HAM/TSP have a higher
proviral load and production of the inflammatorily cytokine IFN-ɣ e TNF-α as compared
to HTLV-1 carriers. While the immune response to HTLV-1 infection is polarized to the
Th1 type, in chronic Schistosoma mansoni infection it is polarization toward the Th2
immune response. In the last five years it has been demonstrated in experimental
models that infection with S. mansoni or injection of S. mansoni antigens modulate
the Th1 inflammatory response involved in some auto-immune diseases, such as: type
I diabetes, rheumatoid arthritis and psoriasis. Our hypothesis is that Schistosoma sp.
antigens are able to down modulate the inflammatory immune response in patients
infected with HTLV-1. Methodology: The Schistosoma antigens Sm29, ShTSP2 and PIII
were added to the cultures of peripheral blood mononuclear cells (PBMC) of patients
infected with HTLV-1 and the levels of cytokines in the supernatants were measure using
the ELISA sandwich method. The antigen Sm29 is a glycoprotein from the tegument of
the S. mansoni adult worm and PIII is a protein obtained from the soluble adult worm
antigen (SWAP). Both induce IL-10 production in vitro by PBMC of chronically infected
individuals. ShTsp2 is a protein from the S. haematobium tegument and the immune
response to this protein in humans is not well known. Results: Compared to the levels
of cytokine in PBMC unstimulated culture, the levels of IFN-ɣ were reduced by the
presence of Sm29 in 8 of 14 patients (mean reduction= 48%; p=0.0019). ShTsp2 was able
to decrease the levels of IFN-ɣ in 7 of 14 patients (mean reduction= 51%; p=0.0058) and
PIII in 6 of them (mean reduction= 39%; p=0.0159). The levels of TNF-α also decreased
after the addition of Schistosoma antigens. In cultures stimulated with Sm29 there was
a reduction in TNF-α production in 4 of 11 individuals (mean reduction= 55%). ShTsp2
was able to down modulate the production of this cytokine in 5 of 11 individuals (mean
reduction= 50%; p=0.035). When PIII was added to the cultures there was a reduction in
TNF-α production in 7 individuals (mean reduction= 71%; p=0.0039). We also evaluated
the levels of IL-10 in the supernatants of PBMC cultures stimulated with the different S.
337
mansoni antigens. The addition of Sm29 and ShTsp2 resulted in an increased production
of IL-10 in 12 of 15 individuals (mean increase= 1113% and 2235%, respectively;
p<0.0001). PIII was also able to increase IL-10 production in 7 of 15 individuals (mean
increase= 45%; p=0.0063). Conclusions: We conclude from these preliminary results that
Schistosoma sp. antigens are able to down modulate the IFN-ɣ and TNF-α production
in peripheral blood mononuclear cells in about 50% of patients infected with HTLV-1.
The down-regulation of inflammatory cytokine production by Schistosoma antigens
may be associated with the augmented IL-10 production. These data may contribute to
the development of strategies to prevent the inflammatory process in HTLV-1 infection.
Financial support- CNPQ (Universal 479417/2008 3) and NIH (R01AI079238A).
338
StUdY Of tHE BaCtERIaL tRanSLOCatIOn tHROUgH tHE IntEStInaL tRaCt In MICE
InfECtEd BY SCHISTOSOMA MAnSOnI
andré de Lima aires; Kedma de Magalhães Lima; thays Miranda almeida; tiago Moreira
alves feitosa; Renata alexandre Ramos Silva; Maria Helena Madruga Lima Ribeiro; Mônica
Camelo Pessoa de azevedo albuquerque; Célia Maria Machado Barbosa de Castro
Universidade Ferderal de Pernambuco, PE - Brasil
Introduction: Schistosomal patients may present impaired drainage by mechanical, irritative,
traumatic, and spoliative actions caused by the obstruction of mesenteric vessels due to the
presence of adult worms. Moreover, spiky eggs of Schistosoma mansoni laid by females in
mesenteric vessels need to overcome the intestinal mucosa to reach the light of intestinal
tract and be eliminated with the feces. However, part of these eggs is retained in intestine
leading to a granulomatous chronic inflammatory reaction. Thus, intestinal epithelium suffers
morphological and physiological alterations. Additionally, during infection by S. mansoni
there is predominantly a Th2 immunological response rather than Th1 causing a relative
immunodeficiency. Because of immunodepression, alteration of microbiota and breaking of
the defensive barrier of intestinal mucosa, the translocation of microorganisms from the
intestinal light to the bloodstream with consequent multiplication and bacterial sepsis may
arise. The present study evaluated the occurrence of bacterial infections in mice infected
by S. mansoni as a result of the translocation of intestinal microorganisms to the peripheral
blood, portal system, spleen, liver, and mesenteric lymphnodes. Methodology: Ten 30-dayage Swiss Webster female mice that were percutaneously infected by 50 cercariae of
Schistosoma mansoni (SLM strain) obtained from Biophalaria glabrata and ten not infected
mice that formed the control group were utilized. Fifty days after infection, it was performed
the individual parasitological test by mean of Kato-Katz method to confirm the infection of
animals. After 90 days of infection, these two groups were sedated for the peripheral blood
collection by caudal puncture. Then, it was performed a midline incision in the xiphoid-pubic
region for the collect of tissue fragments of liver, spleen, and mesenteric lymphnodes of
middle region of the small intestine as well as of portal blood. Samples were added to BHI
and stored at 37ºC for 24 hours for posterior culture on specific mediums. All materials
were collected and cultured with strict asepsis. Results: All animals infected by S. mansoni
presented uniformity in the number of eggs eliminated per gram of stool with average of
192.0 (± 8.76). With respect to the presence of bacteria in culture, not infected animals
did not show positivity on peripheral blood, portal system, liver, and spleen. But, in this
same group, bacterial positivity of 30% in samples of mesenteric lymphnodes was observed.
Considering the infected mice, 90% presented bacterial positivity in mesenteric lymphnodes,
60% in portal system, 50% in liver and spleen, and 30% in peripheral blood. Conclusions:
In the present work, immunosuppression caused by the experimental schistosomiasis
mansoni favored the translocation as well as the migration and multiplication of bacteria on
bloodstream and tissue fragments. Thus, the incapacity that animals infected by S. mansoni
have to avoid the translocation of microorganisms composing the intestinal microbiota is an
important point that needs to be better elucidated.
339
ULtRaSOnOgRaPHIC and LaBORatORIaL EvaLUatIOn Of CHROnIC SCHIStOSOMIaSIS
ManSOnI PatIEntS
ana virgínia Matos Sá Barreto; vinícius Martins alecrim; tibério B. Medeiros; Edmundo
P. a. Lopes; ana Lúcia Coutinho domingues; Sílvia Maria Lucena Montenegro; Clarice
neuenschwander Lins de Morais
Introduction: Schistosomiasis mansoni is a tropical parasite disease that chronically infects
more than 200 million people in developing countries. Schistosoma mansoni worms live in
mesenteric and portal veins of their human host. The chronic clinical manifestations vary
depending on the parasite localization and the parasitic load. The chronic phase may present
intestinal (I), hepatointestinal (HI) and hepatosplenic (HS) forms, with the latter being a
main indicator of schistosomiasis severity. The hepatic lesion is caused by a granulomatous
response to eggs of S. mansoni with subsequent periportal fibrosis, portal hypertension,
splenomegaly, esophageal varices and recurrent hematemesis. Schistosomiasis continues
to be a public health problem in the state of Pernambuco with epidemiological profile
of chronic prevalence in the rural areas and recent cases of acute infection in the coastal
areas. This study objective was to evaluate the ultrasonographic and laboratorial findings
in chronic schistosomiasis mansoni patients of different areas of the state of Pernambuco
selected at the ambulatory of Hospital das Clínicas, Universidade Federal de Pernambuco,
Brazil. Methodology: All the patients were submitted to abdomen ultrasound assessments
according to Niamey protocol and had blood (10ml) samples collect for a complete blood
count and determination of aspartato aminotransferase (AST), alanine aminotransferase
(ALT), alkaline phosphatase (AP), ɣ-glutamil transferase (ɣ-GT), albumin and bilirubin (Bil)
levels. The prothrombin time (PT) and serology for hepatitis B and C were also determined
in every patient. The results have been analyzed through linear regression, Pearson
correlation coefficient and one-way analysis of variance with pairwise T test (p < 0.05 was
considered statistically significant). Results: One hundred and thirteen patients with chronic
schistosomiasis were selected, being 14 (12.5%) I, 36 (31.8%) HI and 63 (55.7%) HS. Their ages
ranged from 18-65 years (Mean ± SD = 48.9 ± 13.6); 46 (40.7%) were male and 69 (59.3%)
female. According to ultrasonography, 9 (8%) patients had pattern A, 5 (4.5%) had pattern B,
11 (9.7%) pattern C, 35 (31%) D, 43 (38%) E and 10 (8.8%) had pattern F periportal fibrosis. The
presence of collateral circulation and recurrent hematemesis were observed in 41 (36.3%)
and 14 (12.4%) of the HS patients, respectively. The longitudinal spleen size correlated with
portal and splenic vein diameter, periportal fibrosis and collateral circulation (r = 0.37; 0.52;
0.52; 0.49, respectively; P < 0.0001) and hematemesis (r = 0.28, P = 0.0018). The portal vein
diameter correlated with splenic vein diameter (r = 0.42, P < 0.0001) and periportal fibrosis
(r = 0.26, P = 0.0066). The splenic vein diameter correlated with periportal fibrosis (r = 0.39, P
< 0.0001). The laboratorial analysis showed HI patients had higher levels of AST, AP and Total
Bil in comparison with I patients (P = 0.0355; 0.0308; 0.0444, respectively). HS patients had
higher levels of ALT (P = 0.036), AST (P = 0.0004), ɣ-GT (P < 0.0001), AP (P = 0.0007), Total Bil
(P = 0.0056), Direct Bil (P = 0.0014), Indirect Bil (P = 0.011); higher PT (P = 0.001) and INR (P
= 0.012); and lower number of Leukocytes (Leu) (P = 0.0028) and Platelets (Plat) (P < 0.0001)
when comparison with I patients. When we compare HS patients with HI, we observed that
340
HS patients had higher levels of AP (P = 0.0308) and Direct Bil (P = 0.0326); and lower number
of Leu and Plat (P < 0.0001). The analysis of ultrasonographical and laboratorial showed a
correlation between the longitudinal spleen size and Bil levels (r = 0.36, P = 0.0055) and
a negative correlation with the number of Leu and Plat (r = - 0.47; - 0.52, respectively; P <
0.0001) and red blood (r = - 0.24, P = 0.0189). And a negative correlation between the portal
vein diameter and the plat count (r = - 0.29, P = 0.0024). Conclusions: Through this analysis,
we observed that the aggravation of disease is followed by alterations in the blood counts
and the levels of hepatic enzymes.
341
UnCOMMOn PRESEntatIOn Of IntEStInaL SCHIStOSOMIaSIS: InCREaSEd
MORBIdItY WItH dIStInCt OUtCOMES and IMMUnOLOgYCaL PROfILES In PatIEntS
attEndIng a tERCIaRY HOSPItaL
Marta guimarães Cavalcanti; Margareth Maria L. gonçalves; Leonardo f. Silva; José Mauro
Peralta; Kalil Madi
Universidade Federal do Rio de Janeiro, RJ, - Brasil
Introduction: Intestinal involvement in schistosomiasis is one of the most common clinical
presentations of Schistosoma mansoni infection in Brazil. It is mostly asymptomatic or
oligosymptomatic but it also accounts for some severe forms of disease as a result of
exacerbated inflammatory response with considerable morbidity in susceptible individuals.
Anti-schistosomal therapy is frequently effective in parasite control and can also interfere
with chronic manifestations in some individuals but others may progress with persistent
inflammatory reaction, profound tissue damage and organ dysfunction in spite of successful
treatment. Herein, we describe two uncommon presentations of intestinal schistosomiasis
that progressed to a non-specific chronic colitis and enteritis responsive to sulphasalazine in
one and a rectal fistula in the other despite anti-schistosomal treatment and immunological
protection against Schistosoma mansoni. Methodology: Retrospective analysis of two cases
admitted at Hospital Clementino Fraga Filho / UFRJ. Results: Case Reports: a 36 year- old male
born in Pernambuco (Brazil) and living in Rio de Janeiro (RJ) for the past 10 years had a history of
aqueous and/or bloody diarrhea for two years. Six months before admission he progressed with
hypochondral pain, increased abdominal volume at the left hypochondrium and mesogastrium,
weight loss and a large palpable spleen at physical examination. Laboratory analysis showed
pancytopenia, hypoalbunemia, mild elevation of hepatic enzymes and negative serology for
HIV, HBV and HCV. Eggs of Schistosoma mansoni were detected in one out of six stool samples.
Anti-Schistosomal IgG1, IgG4 and IgE were reactive (titles: 1, 80; 1, 46; 0, 94, respectively).
Ultrasound showed liver right lobe atrophy and left lobe hypertrophy, increased peri-portal and
peri-vascular fibrosis, thickening of gallbladder wall and splenomegaly. Endoscopy revealed
esophageal and gastric varices. Sigmoidoscopy showed mucosal congestion, ulcerations
and polyps and histopathologycal examination showed loss of colonic mucosal architecture,
glandular hyperplasia, several pseudo-polyps and increased mononuclear cellular infiltration.
Though Praziquantel (PZQ) was effective to control egg excretion, clinical and colonoscopic
follow up demonstrated that persistent diarrhea and no weight gain were associated to severe
chronic enterocolitis albeit the absence of S. mansoni eggs. Sulphasalazine was introduced
and he presented regression of intestinal symptoms and bowel biopsy showed improvement
of colonic abnormalities and absence of eggs in the colonic and/or rectal tissue. After a
two-year follow up, rectal biopsy remained negative for the presence of S. mansoni ova and
serological profile showed IgG1 and IgE titles as 3, 10 and 2, 34, respectively. Case 2: a 32
year-old female natural from Lageado, Sumidouro, RJ, presented recurrent perianal pain and
drainage through a perianal fistula during 12 months. Eggs were detected in stool samples and
PZQ was introduced. After 4 months post-treatment, intermittent drainage persisted. Rectal
biopsy showed only empty egg shells without miracydium and intense fibrotic granuloma
reaction. Conclusions: Exacerbated intestinal inflammatory responses increase schistosomiasis
morbidity even after parasite elimination and despite immunological protective response.
342
antIfIBROtIC aCtIOn Of SILYMaRIn On ExPERIMEntaL SCHIStOSOMIaSIS
Hílton antônio Mata dos Santos; fabiana gonçalves Lino; Carolina Carneiro Rocha; fabíola
Ramos xavier; Letícia Campos da Costa; Moragana t. L. Castelo-Branco; Claudia neto Paiva;
alexandre dos Santos Pyrrho
Introduction: Schistosomiasis is a millennial disease that affects about 200 million people in
the world and it is responsible for an annual mortality of 100 to 250 thousand people. Despite
the existence of effective drugs that destroy the parasite, this infection still causes a great
individual and social impact. Although morbidity is frequently associated with inflammatory
granulomatous reaction and the subsequent liver fibrosis, parasiticide drugs used to treat this
disease are generally unable to reduce the pathological sequelae, despite of some indirect
effect of these drugs mainly on acute phase lesions. The fibrosis caused by schistosomiasis can
evolve to portal hypertension and sometimes fatal bleeding. Therefore, the search for new
strategies able to reduce granuloma formation and fibrosis is quite relevant as an adjuvant
treatment for this disease. The therapeutic effect of products which present antifibrotic,
anti-inflammatory, and antioxidant activities as silymarin can minimize the deleterious effect
of Schistosoma mansoni infection. Object: Investigate the anti-fibrotic effects of silymarin on
chronic phase of murine experimental infection with Schistosoma mansoni. Methodology:
Adult female BALB/c mice were infected with 60 cercariae of Schistosoma mansoni BH strain,
by transcutaneous route. Silymarin was suspended in 1% carboxymethylcellulose (CMC),
being established dosage of 10 mg.kg-1 every 48 h, administered by intraperitoneal route.
Mice were divided in six groups: two groups non-treated (non-infected (N) and infected (I)),
one group infected and treated with 40 doses of CMC (I+CMC 40D), and three groups treated
with 25 (I+SIL 25D) or 40 doses (N+SIL 40D and I+SIL 40D) of silymarin. The mice were observed
for 120 days post-infection and submitted to euthanasia. Liver, spleen, and intestinal tissues
were analyzed and weighed to relate them with body weight. The quantification of eggs in
hepatic and intestinal tissue was performed after digestion of tissue with KOH. To evaluate
the hepatic fibrosis, hydroxyproline was quantified by a colorimetric assay. Results: The
treatment with silymarin in experimental schistosomiasis leads to a decrease of hepatomegaly
in the group that received longer treatment (I+SIL 40D). No differences were observed in
splenomegaly between infected treated and infected non-treated groups. All infected groups
had the similar parasite burden evaluated by number of eggs in the hepatic and intestinal
tissue. The decrease on hepatomegaly observed was associated with significant reduction in
hepatic fibrosis which can result in a diminution of destructive and obstructive lesions of the
intrahepatic vascular system, common in the advance form of schistosomiasis. Conclusions:
The search for treatment that can minimize the deleterious effects of Schistosoma mansoni
infection is essential to accelerate the reversion of established damage due to this infection.
The results observed herein show that the treatment with silymarin leads to a reduction in
liver fibrosis that was associated with a decrease in hepatomegaly. Thus, the silymarin is a
promising drug to reduce the morbidity caused by Schistosoma mansoni.
343
antIgEn-SPECIfIC Igg4 IS a MaRKER Of RESIStanCE tO SCHIStOSOMIaSIS In EndEMIC
aREaS In BaHIa, BRazIL
Joanemile Pacheco de figueiredo; Ricardo Riccio Oliveira; Luciana Santos Cardoso; Maria
Cecilia almeida; diego Mota Lopes; Leda Maria alcantara; Edgar Marcelino Carvalho; Maria
Ilma araujo
Serviço de Imunologia da Universidade Federal da Bahia, BA - Brasil
Introduction: National control strategies to schistosomiasis focusing on mass chemotherapy
with praziquantel have significantly reduced severe pathology. However, reinfections
with consequent morbidities persist despite repeated treatments. In vitro studies have
demonstrated that schistosome larvae are killed in the presence of sera from infected
individuals together with leukocytes from uninfected donors, suggesting that parasitespecific antibody-dependent cellular cytotoxicity (ADCC) plays a key role in parasite
elimination. Subsequent investigations identified the role of protective immunoglobulin E
(IgE), IgA, and IgG antibody isotypes and the participation of eosinophils and mast cells in
orchestrating this attack. Other studies have demonstrated the presence of the inhibitory
isotype IgG4 which block schistosomular killing by competing with the protective isotypes.
Therefore, schistosome induce both protective and antagonistic antibody responses, which
may alter the balance between parasite elimination and immune evasion. We evaluate
serum levels of Schistosoma mansoni-specific IgE and IgG4 in individuals living in an endemic
area of schistosomiasis in Bahia. Methodology: Individuals living in four villages of CondeBA were invited to participate in this study. The participants were submitted to parasite
exams using Kato-Katz method and to blood collection to further measurement of soluble
adult worm antigen (SWAP)-specific IgE and IgG4 using ELISA methods. After that they were
treated with Praziquantel and re-evaluated six and fourteen months after treatment. Results:
Five hundred fifty individuals for about 1000 living in the four villages agreed to participate in
the study. The prevalence of S. mansoni infection in the area was 45% and the mean parasite
burden was 203 ± 338 eggs per gram of feces. The mean level of SWAP-specific IgE and IgG4
were 0.8 ± 0.44 and 1.45 ± 1.48 U/mL, respectively. There was no significant difference in the
mean levels of IgE in individuals with different parasite burden, while the levels of IgG4 were
significantly higher in individuals with high parasite burden (p < 0.05). The ratio between
SWAP-specific IgE/IgG4 was increased in highly infected individuals than in low-infected ones
(01 to 99 eggs/gf =13,2±54,9; n=269; 100 to 199=5±27, n=115; 200 to 400=1,6±4,4, n=18;
200 to 400=0,48±0,69, n=19;> 400eggs/gf=0,33±0,27, n=11). Besides, there was a negative
correlation between the ratio of SWAP-specific IgE/IgG4 and parasite load of S. mansoni
(p<0.005). Regarding the post-treatment evaluation, the parasite burden remained low for
14 months after treatment and strangely it was followed by low levels of SWAP-specific IgE
and high levels of SWAP-specific IgG4 in an evaluation performed six month after treatment.
Conclusions: We demonstrated that, in poor rural area endemic for schistosomiasis in Bahia,
the resistance to schistosomiasis is a phenomenon associated with low levels of SWAPspecific IgG4 and increased ratio between SWAP-specfic IgE/IgG4. Financial support: Johns
Hopkins University and Immunology Service, Federal University of Bahia.
344
CYtOKInE PROdUCtIOn PROfILE aSSOCIatEd WItH PERIPORtaL fIBROSIS In HUMan
InfECtEd WItH S. MAnSOnI
gabriela da Silveira e nunes; Mônica Maria de almeida; andréa teixeira Carvalho; Lúcia alves
de Oliveira fraga; tiago da Costa Morais; giovanni gazzinelli; Elaine Speziali; Olindo assis
Martins-filho; Rodrigo Correa-Oliveira; andrea gazzinelli; alda Maria Soares Silveira
Universidade Vale do Rio Doce, MG - Brasil
Introduction: It has been well documented that in endemic areas, most individuals infected
with Schistosoma mansoni exhibit a chronic relatively asymptomatic infection referred to as
the intestinal clinical form of the disease. However, a small percentage of infected individuals
develop severe clinical forms of the disease, including different grades of periportal fibrosis.
This pathology has identified cytokine response as an important factor in developing the
disease being periportal fibrosis the major, consequence of this process. The goal of the
present study was evaluated the immunological events involved in pathogenesis of periportal
fibrosis, comparing human response from individuals with or without periportal fibrosis,
before and two year after treatment with praziquantel. Methodology: We have investigated
the profile of cytokine synthesis by peripheral blood mononuclear cells (PBMC) from thirtyone volunteers from endemic area infected with S. mansoni, prior to and two years after
treatment. Ultrasonography was carried out to score periportal fibrosis in patients from Caju,
a rural area at Vale do Jequitinhonha, M.G., Brazil, based on the grade of ecogenicity according
to WHO classification. The levels of interferon-gamma (IFN-g), tumour necrosis factor alpha
(TNF-α), and interleukins (IL)-5, IL-13, IL-10, IL-17 and Transforming Growth Factor- β (TGF-β)
were measured in supernatants of PBMC cultures stimulated with soluble antigens from
schistosome eggs (SEA). Cytokines levels were measured using a flow cytometric method
(Cytometric Bead Array). For analyses of cytokine production by PBMC cultured with SEA
data are expressed as stimulation indexes (SI) calculated according to the formula SI= SEA/C
where SEA is stimulated culture and C is control culture. Results: The subjects were classified
into groups according to the presence or absence of pathology and categorized as low or
high cytokine producer. Our results demonstrated that before treatment the proportion of
subjects with high levels of IL-5 (p<0,0001) and IL-17 (p<0,0001) was significantly larger in
the group with fibrosis and it was higher the number of individuals producing high levels of
IFN-g(p<0,0001) and TGF- β (p=0,0147) in the group without fibrosis. After the treatment,
only for TGF- β (p=0,0477) was significant between-groups differences observed in the
percentages of patients exhibiting low and high levels of cytokines. Conclusions: In the
context of cytokine milieu triggered by S. mansoni infection our preliminary data suggest
that IL-5 and IL-17 plays a role in the establishment/maintenance of periportal fibrosis. These
observations strengthen the view that pathology represents a multifactor effect of cytokine
network. Financial support: FAPEMIG; PAPES/CNPq; UNIVALE; CPqRR/FIOCRUZ
345
EndOtHELIaL dYSfUnCtIOn IndUCEd BY SCHIStOSOMIaSIS
Suellen d‘arc dos Santos Oliveira; Luciana Silva do amaral; Luis Eduardo Menezes Quintas;
françois germain noël; Cláudia Lúcia Martins da Silva
Introduction: Schistosoma mansoni is an intravascular parasite that causes a chronic
inflammation related to morphological and functional vascular alterations. The objective
of this study was to evaluate endothelium-related functions such as leukocyte migration,
vascular permeability and function of endothelial nitric oxide synthase. Methodology: All
experiments were approved by local ethics committee (protocol: DFBCICB011). Animals:
Newborn Swiss mice (2.5 - 3 mouths) were infected percutaneously with ~ 80 cercariae of
S. mansoni. Age-matched animals were used as control. Primary culture of endothelial cells:
Mesenteric microvascular endothelial cells were used (Silva et al., 2007. Br. J. Pharmacol., 150:
1-11). Nitric oxide (NO) production stimulated by ATP (100 µm) was measured in first passage
cells using a fluorescent probe (DAF-FM 2.5 µm). Western blotting: Confluent endothelial
cells were lysed and 20 µg of protein loaded on SDS-PAGE gel (7.5% or 12%) and incubated
with eNOS and caveolin-1 (CAV-1) antibodies, respectively. The bands were quantified using
Imagepro program. Leukocyte migration and vascular permeability: Mice were treated
intravenously with Evans blue. The animals were sacrificed and 5 ml of saline (PBS) were
injected intraperitoneally, removed, centrifuged and the pellet resuspended in PBS (1 ml)
to the total and differential count. The supernatant was used to measure the permeability
using the spectrophotometer. Results: Both the expression of the enzyme endothelial nitric
oxide synthase (eNOS) and NO production are reduced in infected animals, suggesting that
the reduction of endothelium-dependent vascular relaxation observed in schistossomiasis
(Silva e cols. 2007. Vasc. Pharmacol. 46: 122-128) is related to the reduction this vasodilator
mediator. The eNOS activity is physiologically inhibited by CAV-1, thus an increase in the
CAV-1 expression could explain the reduction in the production of NO. On the contrary, CAV-1
expression was not affected, showing that the reduced NO production is due to a functional
alteration of eNOS. When compared to control animals, infected animals showed an increase
in the total number of leukocytes in the peritoneum, accompanied by increased vascular
permeability. Since NO also modulates cellular migration (Pober and Sessa, 2007. Nat. rev.
immunol., 7: 803-815.), these data in vivo could also suggest that the endothelial dysfunction
contributes to inflammatory process. Conclusions: In schistosomiasis there is a reduction of
NO synthesis. Moreover, the infection causes endothelial dysfunction that could be related
to increased leukocyte migration and vascular permeability.
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EvaLUatIOn Of IMMUnOLOgICaL PaRaMEtERS aSSOCIatEd WItH InfECtIOn
and REInfECtIOn Of dIffEREnt MURInE StRaInS, C57BL-6 and BaLB-C, WItH
SCHISTOSOMA MAnSOnI
Clarice Carvalho alves; tatiane teixeira de Melo; Patrícia Martins Parreiras; Cristina toscano
fonseca
Introduction: The murine schistosomiasis model is broadly used in the studies of several
aspects of the disease, for instance: pathology, immunology, biology, chemotherapy
screening, and vaccine development. The present work aimed to investigate the cellular
immune responses and the parasite burden after Schistosoma mansoni infection and
reinfection with two different murine strains, BALB/c and C57BL/6, in order to determine
the most appropriate strain to be used in the studies of vaccination in previous infected
individuals. Methodology: BALB/c and C57BL/6 mice were separated into four groups: 1- mice
infected with S. mansoni; 2- uninfected mice; 3- mice reinfected with S. mansoni; 4- mice
infected with S. mansoni and treated. Splenocytes from mice of each group were obtained
on different days post-infection and post-reinfection. The splenocytes were cultured in the
presence of soluble egg antigen (SEA), soluble adult worm preparation (SWAP), soluble
schistosomula antigen (SSA), concanavalin A (positive control) or complete RPMI medium
(negative control). Twenty four and 72h after stimulation, the supernatants of splenocyte
culture were collected and stored at -70ºC for subsequent determination of IL-10, IL-13,
IL-4, IFN-ɣ and IL-17 cytokine production by “sandwich” ELISA. To evaluated parasitological
parameters, individual worm burden was examined 50 days after challenge infection/
reinfection. The eggs in the stool samples were measured by the HPJ technique, and the
number of eggs in the liver and the intestine was estimated following digestion with 10% KOH.
The number of miracidia hatched in the stool samples was also determined. The weight of
the animals was monitored to detect possible changes during infection/reinfection. Results:
No significant difference in the number of eggs or miracidia hatched per gram of feces, and
in the number of females, males and total worms, could be observed, between strains, after
infection or reinfection. Regarding the number of eggs per gram of tissue, no significant
difference was observed between strains after infection, however in reinfected mice, the
number of eggs per gram of liver was significantly higher in C57BL/6 than in BALB/c mice.
Nevertheless, after reinfection, BALB/c mice presented significant increase in the number
of eggs per gram of liver in comparison to BALB/c infected mice. In both strains, infection or
reinfection did not alter animals weight. Regarding immunological evaluation, splenocytes
from C57BL/6 infected mice produced significant amounts of IL-13 cytokine following SSA,
SWAP and SEA stimulation, seven days after infection, and produced also IL-10 in response to
SSA compared to uninfected control. Thirty days after infection, infected mice also responded
to SSA producing IL-13 and IL-10, however, in response to SWAP, unlike in the beginning of
the infection, splenocytes produced IL-10. Sixty days post-infection, significant levels of IL10 and IL-13 were detected in the supernatant of splenocytes culture from infected mice,
when stimulated by antigens of three phases of parasite development. Also significant level
of IFN-ɣ was detected in these cultures stimulated with SEA and SSA. In the beginning of the
infection, BALB/c infected mice produced significant levels of IL-13 and IL-10 in response to
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SSA compared to uninfected mice, and SEA stimulation induced significant production of
IFN-ɣ and IL-13. Fifteen days after infection, significant levels of IFN-ɣ, IL-10 and IL-13 were
detected, in response to the three parasite extract antigens, and significant levels of IL-4
produced in response to SSA stimulation. IL-13 production was also detected in response to
SSA and SEA, 30 days after infection. In addition, there was significant production of IL-10
against SSA at this time point. Sixty days after infection, BALB/c mice produced significant
levels of IL-13, IL-10 and IL-4 cytokines in response to SEA, SWAP and SSA stimulation.
Splenocytes from reinfected C57BL/6 mice, 3 days post-reinfection, produced significant
levels of TNF-α and IFN-ɣ in response to SWAP, when compared with infected/treated mice.
Significant IL-10 production in response to SSA and SWAP, and significant IL-13 production
in response to SSA, were detected in the supernatant of splenocytes culture from C57BL/6
reinfected mice, seven days after reinfection. Thirty days after reinfection, significant levels
of IFN-ɣ were detected in these mice in response to SWAP and SEA stimulation. Regarding
BALB/c reinfected mice, we observed a significant production of IL-4 in response to SSA and
SWAP, and TNF-α in response to SEA stimulation. Conclusions: Although parasite burden after
infection and reinfection did not differ between BALB/c and C57BL/6 strains, the cytokines
production during the course of infection/reinfection differs between both strains, mainly
after reinfection. Financial support: CPqRR/Fiocruz, INEDT.
348
HaEMatOLOgICaL and IMMUnOLOgICaL StUdIES On tHE CO-InfECtIOn Of
SCHISTOSOMA MAnSOnI and PlASMODIuM CHABAuDI USIng a ROdEnt MOdEL.
Pedro Manuel ferreira; Henrique Silveira; ana afonso; Maria amélia afonso grácio
Universidade Nova de Lisboa, Instituto de Higiene e Medicina Tropical, Unidade de
Helmintologia e Malacologia Médica, Unidade de Parasitologia e Microbiologia Médica Portugal
Introduction: Malaria and schistosomiasis are considered to be the two most important
tropical diseases from a public health point of view, due to their associated morbidity and
mortality. Every year, malaria is responsible for more than one million deaths, 300 million
acute cases, 90% of which occur in Africa. Schistosomiasis, endemic in 74 developed
countries results in more than 14000 deaths/year, more than 200 million infected people
and 20 million severe cases, 80% of which in Africa. Infections between Plasmodium spp.
And some helminths are common in areas where malaria is endemic especially in SubSaharan Africa and South America. Several studies showed that an association between
these two diseases, lead to an aggravation of both malaria and schistosomiasis symptoms.
Methodology: In this study we used C57BL/6 mice. We analysed changes in haematological
parameters, such as haemoglobin (Hgb), haematocrit (HCT) and red blood cell (RBC) counts,
as well as changes in Plasmodium parasitemia in a rodent model with an induced mixed
infection. Eight week old female mice were infected with 50 Schistosoma mansoni cercarie
by tail skin penetration. Eight weeks later, they were infected, intraperitoneally, with
one million Plasmodium chabaudi parasitized erythrocytes. 80 mice were divided in four
groups: a) infected with Schistosoma mansoni, b) infected with Plasmodium chabaudi, c)
co-infected with S. mansoni and P. chabaudi and d) uninfected. Mice from these four group
of mice were bled on day 7, 14, 21 and 30, post malaria infection. Spleens were removed
for lymphocyte culture. IL-4 and IFN-g production was measured by ELISA. The study was
repeated in triplicate. Plasmodium parasitemia was evaluated daily by optical microscopy
of Giemsa stained blood films, from individual mice. The haematocytometer Coulter® STKS
was used to obtained the haematological parameters. The haematological parameters were
also obtained in triplicate. Results: The study showed that the co-infected group presented
earlier P. chabaudi parasitemias. Most of them revealed a parasitemia peak at day 5, while
the group infected only with Plasmodium had a parasitemia peak 2 days later. The usual 2830 day peak with malaria infected mice was also seen 2 days earlier in the co-infected mice.
RBC, HCT and Hgb values were similar in malaria infected mice as well as the co-infected,
comparatively with Schistosoma infected mice and uninfected controls. IL4 produced was
almost lower in co-infected group than in S. mansoni infected group. In Co-infected group
IFN-g only can be measured at days 7 and 30 after malaria infection while P. chabaudi
infected group presented always IFN-g production. Conclusions: In comparison with malaria,
co-infected mice presented changes in Plasmodium parasitemia, immune response, total
weight and liver and spleen weight. When we compare the Schistosoma infected group with
the co-infected group statistically significant differences are found in total parasite load,
immune responses and haematological findings. Co-infections did not present significant
hepatomegaly alterations when compared with Schistosoma infections.
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IdEntIfICatIOn Of IMMUnOdOMInantS PROtEIn Of SCHISTOSOMA MAnSOnI
tEgUMEnt
tatiane teixeira de Melo; Rosiane aparecida da Silva Pereira; Paulo Marcos zech Coelho;
Cristina toscano fonseca
Centro de Pesquisa René Rachou, Fiocruz, MG - Brasil
Introduction: The success in the development of vaccines depends on the knowledge of
protective mechanisms, and on the use of antigens and adjuvants able to trigger protective
immune response. In the case of S. mansoni, it is difficult to develop an effective vaccine
against the parasite due to parasite complexity and its ability to regulate the host immune
response. The vaccination as a prophylactic measure, administered alone or with antihelminthic drugs in association, would be important for the control of schistosomiasis. A
potential antigen stratum to compose a vaccine is the S. mansoni schistosomula tegument.
The tegument represents the parasite-host interface and schistosomula is the first stage
to become in contact with the host immune system and the most susceptible life stage.
Recently, our group demonstrated that the schistosomula tegument of S. mansoni is able to
activate dendritic cells (DC) increasing the expression of CD40 and CD86 molecules on the
surface of these cells and the production of IL-12 and TNF-α. Additionally, using the tegument
of schistosomula (Smteg) plus Freund‘s adjuvant in a vaccination protocol, we obtained a
partial protection from 43% to 48% beyond the reduction in the number of eggs in feces and
trapped in the intestine and liver, reducing the number of hepatic granulomas. Thus, the study
of dendritic cell activation and vaccination with Smteg indicated S. mansoni schistosomula
tegument as a promising antigen stratum to be used in vaccine formulations. The aim of
this work is to identify the immunodominant antigens in the schistosomula tegument using
serum from immunized mice in bidimensional Western Blot experiments. These antigens
will be tested in immunization protocols using DNA vaccine in a murine model in order to
validate their protective potential. Methodology: Biomphalaria glabrata snails were infected
with ten miracidia of S. mansoni and 30 days after infection were exposed to artificial light
for 1h to release cercariae. The cercariae were transformed into schistosomula using the
protocol described by Ramalho-Pinto et al (1974). The tegument of schistosomula was
removed by the addition of 0.3 M CaCl2. The solution was centrifuged and the supernatant
containing the S. mansoni schistosomula tegument was again centrifuged at 25000g and
dialyzed in saline. Subsequently, the protein was precipitated with acetone, resuspended in
IEF buffer and the amount of protein was determined by Bradford methodology. To evaluate
the quality of the recovered proteins, one-dimensional electrophoresis was carried out. To
perform 2D eletrophoresis, the samples were applied on IPG strips with nonlinear gradient
separation (NL), pH 3-10. The strips were subjected to rehydration and isoelectric focusing
on the Protean IEF cell at a maximum current of 50 μA and 20°C. The strips were equilibrated
in a solution containing DTT, followed by a second incubation step in the same solution
in which the DTT was replaced by iodocetamida. The molecular weight standard and the
strips were sealed on SDS-PAGE with agarose containing bromophenol Blue to monitor the
electrophoretic run. The gel was stained by the protocol compatible with mass spectrometry
using colloidal coomassie Blue G-250. Another identical 2D-PAGE was transferred to a
nitrocellulose membrane in order to perform the immunoblot. The membrane was blocked
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and then incubated with serum from immunized mice. An anti-mouse total IgG conjugated
to peroxidase was used as the secondary antibody and the reaction was revealed using
Diaminobenzidine (DAB). Results: The serum from mice immunized and protected against
schistosomiasis was able to recognize approximately 40 protein spots, most of them ranging
from 37KDa to 82KDa and isoeletric point from 5 to 8. These proteins will be excised from
an identical 2D-PAGE stained with coomassie colloidal Blue and digested with trypsin. The
purified peptides will be subjected to indentify by (MS/MS). Conclusions: Through proteomic
analysis we intend to identify the components of schistosomula tegument responsible for
the activation of protective immunity against S. mansoni. Immune-proteomic analisys of the
tegument from S. mansoni schistosomula demonstred that approximately 40 protein spots
are recognized by the serum from immunized and protected mice. These proteins may be
involved in the protective immune response induced by Smteg immunization. Furthermore
they represent candidates to be used in an anti-schistosomiasis vaccine. Therefore we will
provide information that can contribute to the development of a vaccine for use in humans,
and consequently in the control of schistosomiasis. Support: CPqRR, INEDT, RIPAG.
351
IMMUnOLOgICaL MOnItORIng Of PatIEntS WItH aCUtE PHaSE Of SCHIStOSOMIaSIS
ManSOnI fOLLOWIng SPECIfIC CHEMOtHERaPY WItH PRazIQUantEL
amanda Cardoso de Oliveira Silveira; Matheus fernandes Costa-Silva; denise SilveiraLemos; Martin Johannes Enk; Cristiano Lara Massara; Maria Carolina Barbosa alvarez; Pedro
Henrique gazzinelli guimarães; Helena Barbosa ferraz; Paulo Marcos zech Coelho; Rodrigo
Corrêa Oliveira; Olindo assis Martins-filho; giovanni gazzinelli; andréa teixeira-Carvalho
Introduction: When individuals non-residents in endemic areas for schistosomiasis have
accidentally contact with water contaminated by cercariae of Schistosoma mansoni develop
the acute phase of infection. Previous studies performed by our group in acute schistosomiasis
showed decrease in the percentage of circulating CD4+ T lymphocytes and CD28+ CD4+ T
lymphocytes as well as increase in the percentage of B-1 lymphocytes (CD5+CD19+) and
HLA-DR+ CD4+ T lymphocytes, as compared to non-infected individuals. The goal of this
current work has been to investigate several immunophenotypic features in circulating
leucocytes of the acute patients before (ACT-BT), one year (ACT-AT I) and two years (ACT-AT
II) after specific chemotherapy with praziquantel. Methodology: The study population (n=10)
acquired the acute phase of S. mansoni infection in the rural district from Igarapé, Minas
Gerais, Brazil. Ex vivo immunophenotyping of peripheral blood leucocytes from healthy
individuals (control group) and patients was performed by flow cytometry. Results: Before
treatment, the analysis of immunological profile demonstrated an increase in the absolute
number of total CD3+ T lymphocytes, mainly owing to increase in CD4+ and CD8+ absolute
counts, as compared to control group. Moreover, circulating leucocytes from ACT-BT showed
a cell activation pattern with increased counts of co-stimulatory molecules as CD28 (T
lymphocytes and granulocytes), CD80 and CD86 (granulocytes) as well as increased counts
of activation molecules such as CD25, HLA-DR and CD69 (T lymphocytes and granulocytes). It
was also observed an increased counts of CD18, CD44, CD54 and CD62L in the T lymphocytes
and granulocytes from ACT-BT group as compared to control group. Regarding the analysis
of chemokine receptors, it was observed an increased expression of CCR3, CCR5 and CXCR3
in the T lymphocytes and granulocytes from ACT-BT group as compared to control group.
After specific chemotherapy, although the majority of immunophenotypic aspects evaluated
returned to the basal levels one year after specific chemotherapy with praziquantel, several
immunological changes have still been maintained, such as absolute increased counts of
CD18, CD28, CD80, CD86, CCR2 and CXCR3 in eosinophils besides of decreased frequency of
CD62L in neutrophils. Two years after specific chemotherapy, the absolute counts of CD18,
CD28, CD80, CD86, CCR2 and CXCR3 in eosinophils and the frequency of CD62L in neutrophils
returned to the basal levels. Conclusions: The maintenance of immunophenotypic changes
one year post-treatment suggest a long impact of specific chemotherapy in the immune
response from patients with acute schistosomiasis following treatment. Two years posttreatment this modified immune profile is not observed.
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IMMUnOMOdULatORY EffECtS Of MEnTHA PIPERITA L. tREatMEnt On MURInE
SCHIStOSOMIaSIS ManSOnI
naiara naiana dejani; Laís Cristina de Souza; vitor Leão; Joice Margareth Rodolpho; Sandra
Regina Pereira de Oliveira; débora Meira neris; Ricardo de Oliveira Correia; vanderlei
Rodrigues; Luis vitor do Sacramento Silva; fabiana Rossetto Morais; Lúcia Helena faccioli;
Heloísa Sobreiro Selistre de araújo; fernanda de freitas anibal
Introduction: Schistosomiasis is a helminthic disease, which causes considerable morbidity
and mortality worldwide. Infection of a suscepyible host with the blood fluke Schistosoma
mansoni results in the release of parasite eggs in the mesenteric and portal venous
system, leading to entrapment of these eggs in the perisinusoidal spaces of the liver and
in the intestinal wall. Tissue localization of antigen-secreting eggs initiates a persistent
immunological insult to which the host responds by marshalling leukocyte recruitment and
activation of inflammatory and immune-mediated responses resulting in granuloma formation
and subsequent fibrosis. The granulomogenic process in schistosomiasis is dependent on
CD4+ T helper (Th) lymphocytes and results in a shift of the immunologic balance from a
Th1 to a Th2 cell type response. Granulomatous inflammation is one of the most complex
cellular immune responses requiring recruitment and activation of a variety of bloodstream
cells incluind monocytes, lymphocytes, neutrophils, and eosinophilic granulocytes. Cell
adhesion between endothelial cells and leukocytes and between leukocytes mutually is
a key event in the orchestration of cellular interactions which lead to development of a
circumscribed inflammatory infiltrate know as a granuloma. The adhesion molecules ICAM-1
and VCAM-1 are both expressed in S. mansoni egg granulomas and these molecules provide
essential immunological interactions not only for the initiation of granuloma formation but
also for the maintenance and modulation of the schistosomal granuloma during chronic
infection. Methodology: The present study examined the role of Mentha piperita L. in
cellular recruitment and the expression of the molecule adhesion ICAM-1 on the surface
of leukocytes. In female mice weighing 18-20g was injected s.c. with 50 cercariae/animal.
Animals were divided in 3 groups: control non-infected, infected non-treated, and infected/
treated by gastric intubation daily during 48 days with 100mg/Kg of Mentha piperita
L. extract diluted in water. Animals were euthanised between the 7th and 48th day after
infection, and the expression of the adhesion molecule ICAM-1 on surface of cells from blood
and peritoneal cavity was determined by flow cytometry. Results: The results showed that
Mentha piperita L. may modulate the expression of ICAM-1, decreasing the expression of
this adhesion molecule on surface leukocytes. Conclusions: Thus, this natural extract may
contribute to decrease leukocytes recruitment and granuloma formation. Support: CNPq
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IMUnOMOdULatIOn Of tHE IMIdazOLInE dERIvatIvE 3-BEnzYL-5-(4-CHLOROaRILazO)-4-IMIdazOLIdInE-2-OnE In tHE PREPatEnt PHaSE Of tHE InfECtIOn BY
SCHISTOSOMA MAnSOnI
Luiz Henrique de Souza teixeira; Juliana Kelle de andrade Lemoine neves; andré de Lima
aires; tiago Moreira alves feitosa; Renata alexandre Ramos Silva; Maria do Carmo alves
de Lima; Ivan da Rocha Pitta; Suely Lins galdino; valdenia Maria Oliveira de Souza; Mônica
Camelo Pessoa de azevedo albuquerque; vlaudia Maria assis Costa
Introduction: Immunologically, Schistosomiasis mansoni is characterized by a dichotomic
pattern in the interleukins synthesis. At the prepatent phase of the S. mansoni infection
is observed an increasing of Th1-cytokines profile followed by the increasing of Th2cytokines profile. This change of immune profile presents direct relation with the
migration of evolutionary forms of worm and the formation of granuloma in response
to the egg antigens in a vertebrate host. In the treatment of schistosomiasis has been
employed praziquantel controlling the morbidity and acting only against the worms. In
the search for new schistosomicidal drugs is very important the pharmacokinetic study
about the immunologic response during the entire course of inflammatory events. In
this purpose, the imidazole derivative 3-benzyl-5-(4-cloro-arilazo)-4-tioxo-imidazolidine2-one (LPSF/PT05) has been exhibited antiparasitic activity against adult worms of S.
mansoni in in vitro studies, but its capacity to modulate the host immune response has
not yet been evaluated.This study aimed to study the modulation of synthesis of cytokines
interleukin-4 (IL-4), interleukin-10 (IL-10), interferon-ɣ (IFN-ɣ) and nitric oxide during the
prepatent phase in mice infected by S. mansoni that were treated with the derivative
LPSF/PT05. Methodology: Forty 30-day-age female mice weighing about 30-36 g were
percutaneously infected with 50 S. mansoni cercariae (BH strain). After infection, animals
were randomly divided in two experimental groups. Each group was subdivided in two
subgroups of 10 animals, being one subgroup treated with the derivative LPSF/PT05 and
the other subgroup was the control group receiving only the drug vehicle. The therapeutic
intervention of subgroups treated with LPSF/PT05 occurred in the dosage of 100 mg/Kg/
day at the 1st day after infection and 30 mg/Kg/day the 14th day also after infection during
five consecutive days. After 15 days post-treatment, groups were sacrificed and spleen
was aseptically collected for the procedure of splenocyte culture. Resuspended cells were
cultured in plates (5x106 cells/ml) with no stimuli (MEIO), in presence of 20 µg of soluble
egg antigens (SEA) or with 5 ng of Con A. Plates were incubated at 37ºC, 5% CO2, and after
24h and 72h supernatants were collected for posterior analysis of the levels of IL-4, IL-10,
and INF- ɣ by mean of ELISA and the levels of nitric oxide was determined using the Griess
reaction. Results were presented by the arithmetic of dosages of the cultures in duplicate
of 8-10 animals per subgroup (+/- deviation pattern). Results: It was observed a significant
increasing (p < 0.01) of IL-4 levels in splenocytes culture of mice treated at the first day of
infection with 100 mg/Kg/day of LPSF/PT05 that were stimulated with homologue antigens
(SEA) and mitogen agents (Con A). With respect to the treatment with LPSF/PT05 at 30 mg/
Kg/day, from 14th day of infection occurred a significant increasing of IL-4 in those mice
stimulated by SEA as compared to controls. The therapeutic scheme and the experimental
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model used in this study demonstrated that LPSF/PT05 did not influence the IL-10 levels
as compared to controls. It was evidenced a non-significant (p > 0.01) decreasing of this
cytokine in animals treated with both studied drug dosages. In the subgroup treated with
100 mg/Kg/day of LPSF/PT05 it was observed an important increasing of IFN-ɣ as compared
to controls without stimuli and Con A. However, the subgroup treated with 30mg/Kg/
day of LPSF/PT05 showed similar levels of IFN-ɣ in controls without stimuli and Con A.
With relation to nitric oxide synthesis, it was only evidenced a non-significant (p > 0.01)
decreasing in the subgroups treated with LPSF/PT05 as compared to its respective control
groups. Conclusions: From our results, we may conclude that the derivative LPSF/PT05, in
the employed therapeutic scheme, exerts a potential stimulator effect of the Th2 response
during the prepatent phase of experimental infection by S. mansoni due to the diminishing
of IFN-ɣ levels and increasing of IL-4 synthesis mainly in the acute phase of infection.
355
InCREaSE In tHE LEvELS Of Igg1 REaCtIvItY In CO-InfECtEd MICE WItH
STROnGYlOIDES VEnEZuElEnSIS dURIng tHE CHROnIC PHaSE Of SCHISTOSOMA
MAnSOnI
Michelle Carvalho de Rezende; núbia Rangel; deborah negrão-Corrêa
Introduction: Parasite infections have high prevalence among residents living in tropical and
sub-tropical areas with deficient sanitation structure, and infection with multiple parasite
species is frequent. Among the species that can be found associated are Schistosoma
mansoni and Strongyloides stercoralis and Necator americanus. Interactions between
helminth parasites in humans can be synergistic or antagonistic, affecting worm burden
and morbidity. However, studies of polyparasitism in human population still rare and
some aspects, such as the importance of immune response, are difficult to be addressed.
Therefore, the development of experimental models would be essential to study the effect
of activation and/or modulation of immune response during multiple helminth infection,
specifically Schistosoma mansoni infected animals in which the immune response changes
over development of the parasite. Therefore, the aim of this experimental work was the
evaluation of parasite burden and immune response in mice infected by Strongyloides
venezuelensis in chronic phase of S. mansoni infection. Methodology: For this experimental
work, Swiss mice, females were separated into 4 experimental groups: 1 - infected with 25
cercariae of S. mansoni, 2 - infected with S. mansoni and co-infected with 700 larvae of
S. venezuelensis during the chronicle phase of schistosomiasis (14 week post-infection),
3 - infected with S. venezuelensis at the same time than group 3, and group 4 composed
by no-infected animals. The parasite burden was estimated during course of the infection,
by counting larvae recovered from the lung, worms in the small intestine and eggs in the
feces of mice infected with S. venezuelensis (groups 2 and 3), while in animals infected
with S. mansoni (group 1 and 2), the number of worms recovered from mesenteric
veins were counted after blood infusion, and the number of parasite eggs retained in
the liver and intestine were estimated after tissue digestion. The number of S. mansoni
eggs eliminated in feces was also estimated after examination of feces samples. Cellular
infiltration in liver, lung and intestine tissue of infected animals were indirectly estimated
by enzymatic activity and level of IgG1 reactive against Schistosoma egg antigen (SEA)
and Schistosoma adult worm antigen (SWAP) in serum samples was estimated for each
experimental group by ELISA-assay. Results: Experimental infection with S. venezuelensis in
mice that were chronically infected by S. mansoni resulted in statistically lower recovery of
S. venezuelensis larvae in lung, adult worms in small intestine and eggs in feces compared
to the parasite burden recovered from mice that were infected only with S. venezuelensis.
Meanwhile, there were no significant differences in the number of worms or eggs of
Schistosoma mansoni quantified in mice co-infected with Strongyloides venezuelensis
compared with mice infected only with S. mansoni. The activity of eosinophil peroxidase
in tissue sample was similar in different infected experimental groups. There was no IgG1
reactivity against SEA antigens during the S. venezuelensis infection. As expected, there
was significant increase in IgG1 reactivity in S. mansoni-infected and in co-infected mice
compared to controls, but no difference was observed in the IgG1-reactivity between these
356
two experimental groups. In contrast, IgG1-reactivity against SWAP elevated after 13 days
of S. venezuelensis infection and the elevation of reactivity observed during the chronicle
phase of S. mansoni infection was significantly higher in co-infected group compared to
S. mansoni single infected. Conclusions: The data suggested a cross-reactivity of SWAPspecific IgG1 with S. venezuelensis antigens, that would be associated the lower infectivity
of the nematode in co-infected mice.
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PROfILE Of LYMPHOCYtE SUBSEtS Of PatIEntS LIvIng In a COMMUnItY Of LOW
EndEMICItY fOR SCHIStOSOMIaSIS ManSOnI In CEaRa-BRazIL
Sara Menezes de Oliveira; Marta Cristhiany Cunha Pinheiro; teiliane Rodrigues Carneiro;
Bruna Cunha de alcantara; Elza Maria Morgado tomaz; felipe fernando da Cruz Inacio;
fernando Schemelzer de Moraes Bezerra; José ajax nogueira Queiroz
Introduction: Human Shistosoma mansoni infection induces a wide variety of host immune
responses. Multiple mechanisms involving cells, antibodies and cytokines have been
described. Lymphocytes subpopulations are important in this process. Human lymphocytes
may be classified in three main populations according to their biological function and
their cell surface antigen expression: T lymphocytes, B lymphocytes and natural killer
(NK) cells. T lymphocytes (CD3+), can be subdivided in functionally different populations.
The most clearly defined of these are helper/inducer T cells (CD3+CD4+) and suppressor/
cytotoxic T cells (CD3+CD8+). T cells are the mediators of cell immunity. B lymphocytes
(CD19+) are the producers of antibodies (plasma cells), they mediate humoral immunity.
NK cells (CD3-CD56+) mediate cytotoxicity against certain tumors and virus-infected cells.
In this study we performed peripheral blood leucocytes count and lymphocyte subtype
phenotyping from patients with parasitic infection and non-infected individuals living in
same region. Methodology: A low endemicity schistosomiasis region was studied in Ceará,
namely Planalto Cajueiro. All participating individuals signed an ethical informed consent.
Parasitological Kato-katz test to determine schistosoma eggs and characterizes the region
endemicity and ELISA test to determine IgG anti-adult worm antigen (Schistosoma mansoni)
were performed to separate groups. To improve the Kato-Katz test sensitivity, three slides
were used for each sample instead of one. It was possible to identify forty patients positive
to schistosoma infection. From negative population, in both tests, fifty one individuals were
chosen at random to form the negative control group. Blood samples were colleted using
tubes with EDTA to carry out hemogram and flow cytometry to characterize lymphocytes
subpopulations. Labeled Monoclonal antibodies anti-CD3, anti-CD4, anti-CD8, anti-CD19 and
anti-CD59, were used to identify them. Results: In pacients with Schistosoma infection (KatoKatz Positive) we obtained the following percentages: CD3+CD4+ = 58,43% + 9,45, CD3+CD8+
= 34,18% + 7,79, CD19+ = 6,57% + 3,18 and CD56 = 19,80% + 6,98. In negative individuals for
Schistosoma infection, control group, we obtained: CD3+CD4+ = 54,98% + 10,74, CD3+CD8+
= 32,72% + 8,60, CD19+ = 7,99% + 4,36 and CD56 = 20,24% + 8,08. Conclusions: There is no
significant variation in the percentage of lymphocyte subpopulations between Schistosoma
infected and no infected individuals. This is probably due to low parasite charge of infected
people living in a region with low endemicity infection. Future researches in regions with
high endemicity to Schistosoma and characterization of the profile of Th1 and Th2 cytokines
may further clarify the involvement of lymphocytes in this disease.
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RELatIOnSHIP BEtWEEn CYtOKInE PROdUCtIOn and HEPatIC MORPHOLOgY In
UndERnOURISHEd MICE InfECtEd WItH SCHISTOSOMA MAnSOnI
Laís amorim Sacramento; andréia ferreira de Barros; fabiana Letícia da Silva; vlaudia Maria
assis Costa; Roni Evêncio de araújo; Eridan de Medeiros Coutinho; Sheilla andrade de
Oliveira; Silvia Maria Lucena Montenegro
Introduction: Schistosomiasis and undernutrition are important public health problems.
Although undernutrition associated with Schistosoma mansoni infection has been studied,
the mechanisms involved in the process of failure in the formation of fibrotic lesions in the
host infected undernourished are not fully understood. Thus, the study of the relationship
between immune response and liver fibrosis in undernourished murine model infected
by schistosomiasis mansoni is still necessary. Methodology: The experimental groups
C57BL/6 mice were divided in well nourished groups submitted to a diet with normal
protein (commercial diet) and undernourished groups submitted to a diet with low protein
(DBR diet) . These groups were infected with 30 cercariae of S. mansoni (LE starin), and
two groups of uninfected mice (control) were fed with the same diets offered to the both
infected groups, respectively. All animals were euthanized after 21 weeks of infection and
conducted to the parasitological, morphological, morphometric, immunological and statistic
studies. Results: The lower values for nutritional parameters were occurred due the deficient
diet ingested by animals. Were observed a smaller number of parasite eggs in the liver of
the undernourished animals (p < 0.05). All mice in the chronic phase of the disease that
were fed with a deficient diet failed to develop pipestem fibrosis. The immunological study
revealed that IFN-ɣ is negatively correlated with the formation of collagen tissue in the liver
of chronically undernourished and infected animals, while IL-13 showed no correlation with
fibrosis in the infected groups, undernourished or wellnourished. Conclusions: The periportal
hepatic fibrosis does not seem to develop in the host undernourished due to changes in
the composition of nutrients in the microenvironment, which interferes in the process of
immunomodulation of schistosomiasis injury. Possibly the INF-ɣ is involved in inhibition of
hepatic fibrosis in undernourished animals.
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SCHISTOSOMA MAnSOnI antagOnIzES TOXOPlASMA GOnDII-IndUCEd IntEStInaL
InfLaMMatORY RESPOnSE In CO-InfECtEd MICE: ROLE Of tnf-aLPHa and IL-17.
Luiz fernando Queiroz; Jacilene Mesquita; Kalil Madi; Heitor S.P. Souza; Marcelo torres Bozza;
Marta guimarães Cavalcanti
Universidade Federal de Rio de Janeiro, RJ - Brasil
Introduction: Imbalances in parasitic – induced immunoresponses are a key determinant
of tissue damage. During parasitic coinfections, host responses may present with severe
tissue damage or in other cases it induces immune protection that reduces pathological
manifestations and disease severity. It is well known that S. mansoni (Sm) and T. gondii
(Tg) induce polar immune responses toward Th2 and Th1, respectively. During S. mansoni
acute infection, T. gondii coinfection induces extensive liver damage mediated by TNF-α,
IL12, IFN-ɣ and MIF. Albeit acute hepatotoxicity, coinfected mice present mild ileitis induced
by T. gondii in response to S. mansoni immunomodulation. Though exacerbated systemic
pro-inflammatory responses correlate well with liver damage in acute S. mansoni and T.
gondii coinfection (SmTg), reversion of T. gondii-induced intestinal pathology by S. mansoni
immunomodulation is not completely elucidated. The aim of this study is to determine the
role of local regulation of TNF-α and IL17 in the intestinal compartments of S. mansoni and
T. gondii coinfected mice. Methodology: Male and female C57BL/6 strain mice (6-10 week
of age) were infected subcutaneously with 50 cercariae of BH strain. A group of S. mansoni
not-infected and 7 week infected was perorally infected with 100 cysts of ME 49 T. gondii
strain. Animals were daily weighted. After 8 days post-T. gondii coinfection, all groups were
sacrificed. Micro-hematocrit was performed in blood samples of non-infected, mono and coinfected mice.. Tissues of all groups were collected for histopathologycal analysis. Levels of
TNF-α and IL17 of all groups were determined by ELISA (Peproteck, Ribeirão Preto, BR) in the
supernatants of intestinal cultures. Significant differences were determined by T-test, ANOVA
or Mann-Whitney when required (software Prism 5, USA). Differences of at least p < 0, 05
are considered significant. Results: Accentuated weight loss was observed in T. gondii monoinfected mice when compared to S. mansoni mono and coinfection (15, 06 ± 7, 5% x – 4,24
± 11,03% and 5,59 ± 8,37 %, respectively; p < 0,05). Mono and co-infected mice presented
significant reduction of hematocrit when compared to naïve mice (Sm 52 ± 1,73,Tg 49,8 ±
1,79,SmTg 42,33± 5,03 x naïve 67,75 ± 1,89, p = 0,0 17) but no differences were detected
among groups. All groups were examined for structural intestinal damage by measuring the
length of the small intestine. T. gondii mono-infection induces significant intestinal shortening
when compared to S. mansoni mono and co-infected mice (17, 23 ± 10, 60% x - 1, 03± 13, 52
% x 6, 44± 7, 11%, respectively. p < 0.05). Histopathologycal analysis of the terminal ileum
showed that S. mansoni mono-infected mice presented intestinal architecture well preserved
and few granulomas at the ileum with or without intact eggs mostly with lymphocyte
accumulation. On the other hand, T. gondii mono-infection produced loss of villous-crypt
ratio, important blunting of villi , intense necrosis and mononuclear infiltration of lamina
propria. In coinfected mice, S. mansoni –induced granuloma did not differ from monoinfected mice. Nonetheless, it presented reduced cell infiltration and less necrosis when
compared to T. gondii mono-infected mice. At the gut, TNF-α levels were similar in mono and
coinfected mice (Sm 27,20 ± 9,12 x Tg 24,23±15,63 x SmTg 39,45± 29,54pg/ml) but IL-17 was
360
upregulated in the supernatants of intestinal cultures of Tg mono-infection and coinfected
mice when compared to Sm monoinfection (Sm 22,55 ± 18,66 x tg 116,86 ±27,30 x SmTg120,90
±13,47 pg/ml, p , 0,05). Conclusions: S. mansoni promotes attenuation of T. gondii induced
intestinal tissue damage in S. mansoni and T. gondii coinfected hosts regardless sustained
pro-inflammatory stimuli mediated by TNF-α and IL-17 at the intestinal compartment.
361
SCHISTOSOMA MAnSOnI tEgUMEnt (SMtEg) MOdULatES tHE ExPERIMEntaL
aLLERgIC aStHMa
Cintia Maria gonçalves da Silva; gardênia Braz figueiredo de Carvalho; fábio a. v. Marinho;
Sérgio Costa Oliveira; Cristina toscano fonseca; Lucila grossi gonçalves Pacífico
Introduction: Allergic inflammations are associated to a Th2 type of immune response that
produce large amounts of IL-4, IL-5 and IL-13. These inflammatory mediators induce IgE
production and eosinophilia. Although immune responses between helminthic infections
and asthma are similar, these diseases have a negative association of occurrence. Some
studies performed in schistosome endemic areas showed low prevalence of allergic
diseases, so helminthes infection might reduce asthmatic manifestations. This modulation
has been associated, both in mice and human, to interleukin (IL)-10 production and to an
increased number of T regulatory cells. The schistosomulum is the first parasite stage to
get in contact with the host immune system activating the antigen presenting cells and
promoting the B and T lymphocytes differentiation. Smteg (schistosomulum tegument)
can activate dendritic cells leading to increase the CD40 and CD86 expression and the IL10, IL-12 and TNF-α production. Many factors may be acting to inhibit the allergic asthma
induction in mice associated to schistosomiasis, which may involve both innate and adaptive
immune responses. Our goal is to investigate the mechanisms involved in the immune
modulation of asthmatic response after Smteg intraperitoneal injection, aiming at finding
new biomolecules to be used in asthma treatment. Methodology: Cercariae of Schistosoma
mansoni were mechanically transformed into schistosomula, and the tegument was removed
with CaCl2 0.3M by vortex agitation. The detached membrane sample was separated from
denude bodies by centrifugation. The supernatants were pooled, centrifuged and the pellet
was dialyzed against physiologic saline 1.7%. Balb/C mice were divided in three groups (n=5)
PBS, ASTHMA and SMTEG/ASTHMA. Before the experiments beginning, mice were wormed.
All groups, were subcutaneously primed twice with 10µg of ovalbumin chicken-egg (OVA)
associated with alum, with a 15-day-interval. One week after the first OVA inoculation, mice
from SMTEG/ASTHMA group were intraperitoneally immunized with 25µg of Smteg. The
ASTHMA groups were challenged with OVA aerosol (twenty minutes daily, for five days) to
develop asthma, one week after the second immunization. Mice were euthanized twenty four
hours after the last aerosol challenge. Bronco-alveolar lavage (BAL) was performed and the
lungs were collected to measure cytokines and chemokines by ELISA, an immunoenzymatic
method. Results: We analyzed the number of eosinophils in bronco-alveolar lavage (BAL)
and the levels of IL-4, IL-10, CCL2, and CCL3 in lung tissue. The number of eosinophils was
significantly higher in ASTHMA group when compared to PBS group. SMTEG/ASTHMA group
presented a reduction on eosinophils numbers and also high levels in interleukin 10 (IL-10)
production when compared to ASTHMA group. No statistical differences were observed in
IL-4, CCL2 and CCL3 production among these groups. IL-5, CCL5 and CCL11 still need to be
evaluated. Conclusions: Smteg intraperitoneally modulated the number of eosinophils and it
was associated with an increase of IL-10 production.
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tHE ROLE Of MaCROPHagE MIgRatIOn InHIBItORY faCtOR (MIf) In SEvERE
SCHIStOSOMIaSIS
Jailza Lima Rodrigues; Juliana froeseler fittipaldi; adriana fernandes; Paula duarte Eschenazi;
fernanda S. Costa; Emília Souza araújo; Michelle Carvalho Rezende; José Roberto Lambertucci;
Carlos Maurício antunes; Mauro Martins teixeira; deborah negrão-Corrêa
Introduction: Schistosomiasis is a chronicle and debilitating disease that affects over 200
million people worldwide. Studies in murine model and in human population indicated
that the extension and intensity of the collagen-rich granulomatous response against the
parasite eggs trapped in the liver and intestine can be associated with the development of
severe schistosomiasis. Therefore, factors that influence the induction and/or modulation
of the immune response against parasite egg antigens would be determinant in evolution
of severe schistosomiasis. The pro-inflammatory cytokine designated as macrophage
migration inhibitory factor (MIF) plays a central role in the induction and regulation of
inflammatory responses, through activation of lymphocytes, macrophages and eosinophils.
During Schistosoma mansoni infection, there are reports of increased production of MIF
and the importance of this cytokine to migration and survival of eosinophils into the
granulomatous inflammation. However, the participation of MIF in the development/
modulation of the severe pathology in schistosomiasis was not defined and it was the aim
of this study. Methodology: Balb/c mice genetically deficient in MIF production (MIF-/-)
and non-deficient Balb/c mice (WT), were infected subcutaneously with 25 cercariae/mice
and the liver pathology was evaluated through histopathologic and morphometric analysis
during the acute (8 weeks) and chronicle stage (14 weeks) of the experimental infection.
Moreover, cellular infiltration in liver was indirectly analyzed through enzymatic activity,
specifically the eosinophil peroxidase (EPO) and N-acetylglucosaminidase (NAG) level, and
the collagen content was indirectly determined by quantification of hydroxyproline. Liver
samples of infected mice from both experimental groups were also used to quantified
local production of IL-4, IL-13, IL-10, IL-17, TNF-α and IFN-ɣ cytokines using ELISA-assay
commercially available. In parallel, MIF concentration was determined by ELISA-assay in
plasma samples of 97 people from an endemic area of schistosomiasis, in Padre Paraiso,
northeast of Minas Gerais. All the participants had S. mansoni infection confirmed by fecal
examination and were submitted to clinical and ultrasound examination to define the
disease severity. Results: Data obtained in mouse experimentally infected with S. mansoni
showed no difference in parasite burden (adult worms and eggs) between infected mice
from both experimental groups, but the granuloma formed in liver of MIF-/- infected
mice showed larger cellular infiltration and size compared to non-deficient infected mice.
Enzymatic activity confirmed increase level of liver eosinophil peroxidase during the acute
phase and NAG in acute and chronicle phase of the infection. Moreover, the increase of
hydroxyproline in the liver of MIF-/- infected mice was significantly lower than the level
detected in WT infected mice, indicating that collagen deposition in the liver of MIFdeficient mice was lower. Although the cytokine level detected in liver of infected animals
was similar during the acute phase of schistosomiasis, local production of type 2-cytokine,
IL-4 and IL-13, was lower and the level of IFN-ɣ was higher in MIF-/- chronically infected
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mice compared to WT. Moreover, significant higher level of MIF was detected in plasma
sample of infected people that had larger portal vein diameter and thicker portal vein
wall compared to infected people that show no liver alteration. Conclusions: The results
indicated that increased production of MIF is associated with severe schistosomiasis, due
to larger cellular infiltration in liver granuloma and collagen deposition.
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aLBUMInURIa In SCHISTOSOMA MAnSOnI-InfECtEd IndIvIdUaLS
Milton Cezar Compagnon; Laís danielle Ribeiro de Melo; Carmem de Castro Chaves; ana Lúcia
Coutinho domingues; ana durce Oliveira da Paixão
Introduction: Immunological response has been correlated with increased endothelial
oxidative stress that may lead to microalbuminuria. In its turn, microalbuminuria has been
associated with later development of cardiovascular disease. We investigated whether
patients with hepatosplenic schistosomiasis, infected with Schistosoma mansoni (SM)
parasite, show microalbuminuria. Methodology: Albuminuria, obtained from spot urine
indexed to urinary creatinine, was investigated in 54 subjects showing hepatoesplenic
fibrosis due to SM infection, the SM group and, in 30 control subjects, the C group. Inclusion
criteria required nonhypertensive, nondiabetic and nonsplenectomyzed subjects. Urinary
albumin and creatinine were measured by nephelometry (Behring) and standard colorimetric
method, respectively. The protocol was approved by the local Ethical Committee. Results:
Patients exhibited the following clinical characteristics: mean age 48.8 ± 10.9 years (men,
n = 32) and 42.0 ± 13.6 years (women, n = 22), mean arterial pressure 98.8 ± 8.4 mmHg
(males) and 90.1 ± 12.2 mmHg (women); albuminuria 5.1 ± 3.4 mg / g creatinine (men,
n = 22) and 5.1 ± 3.4 mg / g creatinine (women, n = 14). The control subjects evaluated
showed the following clinical characteristics: mean age 38.8 ± 9.0 years (men, n = 7) 41.6 ±
13.3 years (women, n = 23), mean arterial pressure 97.2 ± 8.9 mmHg (males) and 89.2 ± 9.1
mmHg (women); albuminuria 5.7 ± 3.4 mg / g creatinine (men, n = 6) and 18.9 ± 6.2 mg /
g creatinine (women, n = 22). From the 54 evaluated patients, 3.7 % (1 man and 1 woman)
presented microalbuminuria (values between 30 and 299 mg/g creatinine). Similarly, among
the control individuals, 6.6 % (1 man and 1 woman) showed microalbuminuria. On the other
hand, 29.6 % of SM group (9 men and 7 women ) had undetectable albuminuria, while 66.6
% of SM group (22 men and 14 women) showed normal levels of albuminuria. Conclusions:
Although the possibility of high incidence of cardiovascular disease in individuals infected
with SM should not be ruled out, our preliminary data show that the more advanced stages
of Schistosoma mansoni infection did not appear correlated with microalbuminuria.
365
CHaRaCtERIzatIOn Of antI-InfLaMMatORY aCtIvItY Of a nEW 4,5-dIIdROISOxazOL
dERIvatIvE aS a POtEntIaL COMPOUnd tO dECREaSE tHE SCHIStOSOMaL
gRanULOMa fORMatIOn
amanda Roberta Revoredo vicentino; anderson Mendonça amarante; vitor Coutinho
Carneiro; Carlos aleberto antunes; Cláudia farias Benjamim; alcino Palermo de aguiar;
Marcelo Rosado fantappié
Introduction: Schistosomiasis is a serious global helminthic disease, in which the main
immunopathology consists of a granulomatous and fibrosing reaction against tissue-trapped
parasite eggs. Granuloma formation is controlled and modulated by several cell types and
protein interactions, primarily CD4+ T-cells, Th1 cytokines and cell adhesion factors. It is well
established that the Th1 cytokines like TNF-a plays a central role in circumoval granuloma
formation and when infected mice were treated with monoclonal antibody anti-TNF-a,
occur a significant decrease of the granuloma area. Thus, inhibition of cytokine activities
in this disease could be remarkably successful. Therefore, anti-cytokine agents that target
biosynthetic pathways of pro-inflammatory cytokines would offer an attractive alternative to
the treatment of inflammatory diseases as schistosomiasis. Recent data showed the capacity
of VGX-1027 [[(S,R)-3-phenyl-4,5-dihydro-5-isoxasole acetic acid] to inhibit the increase of
circulating levels of TNF-α in lipopolysaccharide (LPS)-challenged mice. In our work, we first
characterized the anti-inflammatory activities of a new 5-(4-pyridil)-4,5-dihydroisoxasole
derivate. Following this characterization, we have started to investigate the anti-inflammatory
roles of this new compound in infected mice and evaluate its contribution in the decrease
of granuloma formation. Methodology: MTT assay for cell viability – Peritonial macrophages
were plated at a density of 100000 cells/well in 96-well plates. To determine the appropriate
concentration of DIC, which showed no effect on cell viability, cytotoxicity studies were
performed at 24 h following treatment of cells with various concentrations of DIC. Viabilities
were determined using colorimetric MTT assays, as described previously. TNF-a and IL-1β
assay - Peritonial macrophages were pretreated with DIC for 2 h and then stimulated with
LPS (100 ng/mL) for 4 h. Levels of TNF-a and IL-1β in the supernatant of the culture were
quantified using ELISA kits (PEPROTECH). Nuclear extraction and electrophoretic mobility shift
assay (EMSA) - RAW 264.7 macrophages cells were plated in 6 well-plates (2,0 x 106 cells),
and treated with DIC (150 and 200 μM), stimulated with LPS (1μg/mL) for 1 h, washed once
with PBS, scraped into 1 ml of cold PBS, and precipitated by centrifugation. Nuclear extracts
(10 μg) were mixed with 20 ng double-stranded NF-kB oligonucleotide 5’-AGTTGAGGGGACTTTCCCAGGC-3’ end-labeled with [y-32P]-ATP. Binding reactions were performed at 37 ºC
for 30 min in 30 μL of reaction volume. DNA–protein complexes were separated from the
unbound DNA probe on native 5% polyacrylamide gels at 200 V in 0.5x TBE buffer. Gels were
vacuum-dried for 1 h at 80 °C and exposed to X-ray film at -70 °C for 24 h. Cell migration in
vivo – Balb/c Mice were treated by intraperitonial injection with DIC (5 mg/kg) and vehicle
(DMSO 2,4%) for 30 minutes and then stimulated with sodium thioglicolate (3 %) for 4 hours.
Peritonial washing were obtained with 3 mL of saline. Total white blood cells count was
carried out by using Turk’s solution and Neubauer haemocytometer. Differential leukocyte
count was performed in citospin using hematoxilin-eosin. Results: MTT assays showed that
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treatment of macrophages with DIC at 150, 200, or 250 μM for 24 h did not induce any
cytotoxicity at the concentrations used. Our data showed that the compound named DIC
significantly reduced the release of TNF-α and IL-1β from murine macrophages stimulated
in vitro with LPS. Since NF-kB activation is required for the expressions of TNF-a, and IL1β by LPS, EMSA assay were performed to determine whether the anti-inflammatory effect
of DIC involves the NF-kB pathway. Treatment with LPS significantly increased NF-kB–DNA
binding, whereas this binding was significantly abolished by DIC (150 and 200 μM). In vivo,
DIC inhibited the migration of neutrophils to the peritoneal cavity of mouse stimulated with
sodium thioglycolate. Conclusions: These results strongly suggested the role of one new
5-(4-pyridil)-4,5-dihydroisoxasole derivate (DIC) as an anti-cytokine mediator.
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CHaRaCtERIzatIOn Of tHE gaMMa-SECREtaSE COMPLEx In SCHISTOSOMA MAnSOnI
and tHE EffECtS Of ItS InHIBItIOn On PaRaSItE dEvELOPMEnt
Lizandra guidi Magalhães; William de Castro Borges; Renata guerra Sá; Carla Botelho
Machado; Enyara Rezende Morais; Érika Bueno de Carvalho Moreira; Cláudia Sossai Soares;
Olinda Mara Brigatto; Elenice aparecida Macedo; vanderlei Rodriges
Introduction: Proteases have been shown to play an important role during the life-cycle of
S. mansoni due to their influence on parasite differentiation, growth, and migration. The
gamma-secretase complex is an assembly of at least four individual proteins: PS (presenilin),
Nicastrin, APH-1 (anterior pharynx-defective 1) and PEN-2 (Presenilin enhancer 2). Although
the exact function of the complex has not been fully addressed, the PS component is an
aspartyl protease responsible for intramembrane proteolysis. A variety of molecules
have been described as membrane targets of the gamma-secretase complex, including
the Notch receptor. The gamma-secretase complex is highly conserved and ubiquitously
present in metazoans, however, little is known about its function in invertebrates. Given
that no information is available regarding its role in parasites, this subject is open for
exploration and was encouraged by studies in Caenorhabditis elegans in which the link
between gamma-secretase complex and embryogenesis has been established. The purpose
of this work was the molecular characterization and the expression profile of the gamma
secretase complex in S. mansoni. In parallel, the in vitro effects of a synthetic inhibitor of the
complex upon parasite pairing and egg development were reported. Methodology: Aiming
the identification of homologue sequences coding for the gamma secretase complex in S.
mansoni, orthologues from Homo sapiens, Drosophila melanogaster and C. elegans were
retrieved and used as query sequences to mine the S. mansoni database through BLASTP
searches. SmPS, SmNicastrin, SmAPH-1 and SmPEN-2 transcript levels were evaluated during
the life cycle of S. mansoni and quantified relative to alpha-tubulin using real-time PCR. For
inhibition assays adult worm pairs were incubated in the presence of 10 to 100 µM DAPT
(N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester) a gamma secretase
inhibitor, in RPMI at 37ºC/5%CO2. Parasite pairing, egg production and embryo maturation
were monitored using an inverted microscope at every 24 h, during 120 h. Results: A detailed
search of the S. mansoni genome database identified putative sequences coding for the
gamma-secretase complex in this parasite. Quantitative PCR analysis revealed the respective
transcripts in all of the investigated stages. Particularly, the data suggested that the transcripts
are abundantly expressed in the egg. Moreover, the inhibition studies revealed that DAPT,
at the lowest concentration, reduced egg laying and aborted embryo development in vitro.
Conclusions: Our results demonstrated that gamma secretase complex may be a role in S.
mansoni development. We believe that a better understanding of the activity and function
of the gamma-secretase complex in S. mansoni could provide an exciting approach towards
compromising parasite development within the vertebrate host, wherein egg-laying is the
main factor associated to liver disease. Financial Support: FAPESP, CAPES, FAEPA.
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CHROMatIn REgULatIOn In SCHIStOSOMES and HIStOnE-MOdIfYIng EnzYMES aS
dRUg taRgEtS
Raymond J. Pierce; Stéphanie Caby; florence dubois; Julien Lancelot; Jacques trolet; Céline
Cosseau; Christoph grunau; guillaume Mitta; Luiza f.a. almeida; Leila nahum; guilherme
Correa de Oliveira
CIIL, Inserm U1019, CNRS UMR 8204, Institute Pasteur de Lille - France
Introduction: Histone modifying enzymes (HME) are central actors in the regulation of the
epigenetic modification of chromatin and aberrant epigenetic states often associated with
cancer led to interest in HMEs as targets for therapy. Among these the histone deacetylases
(HDAC) have been the subject of intense study and a number of HDAC inhibitors (HDACi) are in
clinical trials. HDACs deacetylate acetylated lysine residues in a variety of proteins, including
histones, but also transcription factors and cofactors, as well as non-nuclear proteins such
as tubulin. Of the three main classes of HDACs in eukaryotes, enzymes of classes I and II
share similar catalytic domains and a Zn2+-dependent mechanism, whereas the class III
enzymes, the sirtuins, are phylogenetically unrelated and have an NAD+-dependent catalytic
mechanism. Broadly, HDACi induce cell death in cancer cells via apoptosis but they can also
act on the cell cycle, on tumour angiogenesis or via the regulation of host cell responses.
HDACi have also stimulated interest as anti-parasitic drugs and have been tested against P.
falciparum, Toxoplasma gondii and the major kinetoplastid parasites. We have characterized
and studied the HDACs of Schistosoma mansoni and have shown that HDACi cause the
death of S. mansoni larvae and adult worms in vitro via chromatin hyperacetylation and
the induction of apoptosis. Methodology: S. mansoni HDACs were identified by mining the
genome sequence and full-length coding sequences were validated using RACE-PCR. Their
identity was verified by phylogenetic analysis and their expression during the parasite lifecycle was determined using quantitative RT-PCR. The functional activity of selected HDACs
was tested using reporter gene assays in mammalian cell lines. The effect of HDACi was
determined by measuring their effect on the viability of schistosomula and adult worms and
the induction of apoptosis assayed by the TUNEL method and the induction of caspase 3/7
activity. The overexpression of HDAC target genes after HDACi treatment was determined
using qRT-PCR and correlated to hyperacetylation of the corresponding proximal promoters
using quantitative chromatin immunoprécipitation (qChIP). Native acetylation of H3K9
on the promoters was determined using ChIPSeq. Results: S. mansoni possesses 3 class I
(HDAC1, 3 and 8), 4 class II (HDAC4, 5, 6 and 10) and 5 class III (Sirt1, 2, 5, 6 and 7) HDACencoding genes. Several of the corresponding coding sequences, including SmHDAC8 and
SmSirt1 possess insertions in their catalytic domains compared to mammalian orthologues.
Functional assays show that SmHDAC1 represses gene transcription in reporter gene assays
and that SmSirt1 potentiates the activity of the transcription factor FoxO, indicating the
conservation of HDAC functions in S. mansoni. Treatment of schistosomula or adult worms
with HDACi such as TSA or SAHA (inhibitors of classes I and II) or sirtinol (a sirtuin inhibitor)
induces the death of both larval (schistosomula) and adult worms and this is preceded in the
larvae by the induction of apoptosis as measured by TUNEL staining and the increase in the
activity of caspase 3/7. Moreover, such treatments induce a rapid increase in the general
level of histone acetylation, particularly of H4. This in turn correlates with the overexpression
369
of certain genes, including those encoding caspases 3 and 7. ChIPSeq analysis showed that
H3K9 acetylation of the caspase 7 promoter in untreated parasites was very low. Finally,
qChIP analysis shows that the proximal promoter of caspase 3, and in particular of the
caspase 7 gene show hyperacetylation of histone H4 after HDACi treatment. Conclusions:
These results lead us to consider that schistosome HDACs, as well as other HMEs, are
promising targets for the development of new drugs against schistosomiasis. To this end,
a project (SEtTReND) supported by funding from the EC has been initiated, with the aim of
characterizing the HMEs of S. mansoni, particularly those involved in histone acetylation/
deacetylation and methylation/demethylation. These enzymes will be validated as targets
and specific inhibitors will be identified against selected enzymes that could be candidates as
lead compounds for drug development. Financial support: EC (FP7-Health); Inserm-Fiocruz;
Grant ANR-07-BLAN-0119-02
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CLOnIng and PURIfICatIOn Of CItOPLaSMatIC fORM Of tHE EnzYME
SERInEHYdROxIMEtHYLtRanSfERaSE (SHMt) fROM SCHISTOSOMA MAnSOnI.
angela Maria fala; alexandre Cassago; Ricardo de Marco; glaucius Oliva; Richard garrat;
Humberto d´Muniz Pereira
IFSC - USP, SP - Brasil
Introduction: Schistosoma mansoni is one of the parasitic species responsible for the disease
schistosomiasis. In 2005, theWorld Health Organization estimated that approximately 200
million people were infected with this particular species. Morbidity due to S. mansoni
includes hepatosplenomegaly, liver fibrosis and ascites, and as many as 130,000 individuals
die per year from haematemesis due to related portal hypertension. The disease is associated
with a chronic and debilitating morbidity manifested by its consequences including cognitive
impairment, lassitude, and growth stunting. The enormously active oviproduction of
schistosomes implies very active DNA and RNA biosynthetic pathways. The parasite posses
both pyrimidine and purine salvage pathways and also “de novo” pyrimidine pathway, that
together with the Thymidylate cycle supply the worm with DNA and RNA precursors. The
enzyme serinehydroximethyltransferase is an component of the Thymidylate cycle, which
converts tetrahydrofolate + L-serine into 5,10-methylenetetrahydrofolate + glycine supplying
the thymidylate synthase with one carbon unit needed to the conversion of dCMT in dTMP.
Methodology: It was identify two isoformas for the SHMT enzyme in the S. mansoni genome,
one mitochondrial (Smp_144570) and other citoplasmatic (Smp_179710), these isoforms
codifies an protein with 504 and 458 residues respectively, and shares 49% of sequence
identity. The gene for citoplasmatic from of MTAP was amplificated using enriched cDNA
library. The cloning was done in pET28a-SUMO expression vector and confirmed by colony
PCR. E. coli BL21-CodonPlus (DE3) was used to express the SHMT, in 2XTY medium supplied
with 50 μg/mL kanamycin and 10μg/mL chloramphenicol. The expression was induced with
300μM/mL IPTG for three hours, the protein was purified with affinity chromatography
technique using Talon resin (Clonetech), yielding 36mg/L of culture. The fusion protein
SUMO-SHMT was cleavaged using SUMO protease. The SHMT was further purified using
size exclusion chromatography in SUPERDEX200 16/60 column. Results: The result of cloning
was confirmed by DNA sequencing. The expression and purification were visualized using
SDS-PAGE. The SHMT was dialyzed and using in robotic crystallization trials. Conclusions: This
project belongs to the Puri-pyrimidome Project that aims to obtain all the structures and
kinetics constants of all enzymes involved in nucleotide metabolism of S. mansoni.
371
dEvELOPMEntaL REgULatIOn Of tHE SUMO PatHWaY In SCHISTOSOMA MAnSOnI
Roberta verciano Pereira; fernanada J. Cabral; Liana K. Janotti Passos; vanderlei Rodrigues;
William Castro Borges; Renata guerra de Sá
Universidade Federal de Ouro Preto, MG - Brasil
Introduction: Post-translational modifications (PTMs) of proteins dependent on ubiquitin or
Ub-like molecules, such as SUMO, ISGN15 and Nedd8 play regulatory roles in distinct cellular
pathways. The SUMO (Small-ubiquitin-modifier) dependent modification participates in
a variety of cellular functions including gene expression regulation, nuclear location and
signal transduction. Conjugation of SUMO to proteins occurs through mechanisms similar to
ubiquitination, involving the E1 (SUMO activating-) and E2 (conjugating-) enzymes, plus a third
named E3 ligating enzyme that confers target substrate specificity. Although this type of PTM
has been extensively investigated in higher eukaryotes, little is known about the importance
of SUMO modification in lower organisms. In this context, we have previously reported the
existence of two SUMO paralogs in S. mansoni and demonstrated the attachment of SUMO
to parasite cytosolic and nuclear proteins by Western blotting. Based on the evolutionary
conservation of the SUMOylation components, we hypothesize that this system might
contribute to proteome regulation and therefore should constitute an essential mechanism
for parasite development and survival within its alternating hosts. In this study we measured
the levels of mRNA expression of the SUMO machinery by qRT-PCR using cercariae, adult
worms and mechanically transformed schistosomula (MTS) to infer participation of SUMOdependent modification during cercariae to schistosomula transition. Methodology: S.
mansoni LE strain was maintained by routine passage through Biomphalaria glabrata snails
and BALB/c mice. Adult worm parasites were obtained by liver perfusion of mice after 50
days of infection. MTS cultivated in vitro for 3,5 h, 1, 3, 5 and 7 days were prepared using a
standard protocol. Total RNA from adult worms, cercariae and schistosomula was obtained
using a combination of the Trizol reagent (GIBCO-BRL) and chloroform for extraction, and
then purified on column using the “Purelink Micro-to-Midi Total RNA purification system”
(Invitrogen). The cDNAs encoding for constituents of the SUMO pathway (SmUbc9,
SmRanBP2, SmPIAS, SmSENP1 and SmSENP7) were obtained by PCR amplification. Reversetranscribed cDNA samples were used as templates for PCR amplification using SYBR Green
Master Mix UDG-ROX® (Invitrogen) in a 7500 Real Time PCR System (Applied Biosystems).
Specific primers for S. mansoni α-tubulin were used as a constitutive control. For all
investigated transcripts three biological replicates were performed and their gene expression
normalized against the α-tubulin transcript according to the 2−ΔCt method, using the
Applied Biosystems 7500 software. Statistical analysis was performed using GraphPad Prism
version 5.0 software package (Irvine, CA, USA). Normality of the data was established using
Kolmogorov-Smirnoff test and one way analysis of variance (ANOVA). Tukey post hoc tests
were used to investigate significant differential expression of SUMO enzymes throughout the
investigated stages. In all cases, the differences were considered significant when p values
were < 0.05. Results: Our analyses demonstrated the presence of nine molecules responsible
for SUMOylation of target substrates in S. mansoni. Among these, three belong to the E1like family, one E2-like, two E3-like ligases and two desumoylation enzymes. Quantitative
RT-PCR showed that SmAos1 and SmUba2 heterodimers have similar expression profiles in
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all investigated stages. These genes and SmUbc9 were highly transcribed in MTS-3,5h (about
4-fold) when compared to cercariae and schistosomula stages (MTS-1, 2, 5 and 7 days). The
expressions of the E3 ligases, SmPIAS-like and SmRanBP2-like, were 3 and 15-fold higher in
3,5h MTS when compared to their levels in cercariae and adult worms. In agreement, SENP
1/7 were 5-fold more expressed in MTS-3,5h when compared to their levels in cercariae
and adult worms. Conclusions: The observed differential gene expression for SUMOylation
pathway components during cercarie to schistosomula transition suggests a significant
role for SUMOylated proteins during parasite development. Identification of the modified
target substrates will contribute to understand the molecular mechanisms employed by the
parasite during infection of the vertebrate host. Support by FAPEMIG
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SEQUEnCE anaLYSIS and Rna IntERfEREnCE StUdIES Of tRanS-SPLICEd tRanSCRIPtS
fROM SCHISTOSOMA MAnSOnI
Marina de Moraes Mourão; nathalie dinguirard; francisco Pereira Lobo; francisco Prosdócimi;
timothy P. Yoshino; glória Regina franco
Introduction: Spliced leader trans-splicing is a post-transcriptional RNA processing
event in which spliceossomal transference of a short RNA sequence (SL) occurs. The
donor sequence (5’ end of a specialized non-mRNA molecule [SL-RNA]) is joined to an
unpaired splice-acceptor site on a particular set of pre-mRNA molecules. To date, very
little is known about this mechanism, especially in the trematode parasite, Schistosoma
mansoni, in which 30% of transcripts are believed to undergo trans-splicing. The transsplicing mechanism in protozoan and metazoan parasites can also represent a potential
target for the development of drugs that could inhibit this process and consequently
the development of these parasites in their hosts. In a previous study, we presented the
analysis and construction of four SL enriched cDNA libraries for different stages of the S.
mansoni life cycle. At the present study, we attempt to silence the trans-splicing mechanism
in primary sporocysts by RNA interference knockdown. Methodology: Previously, four
cDNA libraries were constructed and cloned using PCR amplified products derived from full
length SL transcripts of S. mansoni adult female and male separate genders, schistosomula
and eggs. cDNA clones were randomly selected from the libraries, sequenced and resulting
sequences were edited, assembled to generate contigs and annotated against several DNA,
protein and InterPro databases. Functional annotation of biological processes is being
conducted using the BLAST2GO software. Here, small interfering RNAs (siRNA) directed to
the SL sequence were synthesized and used to treat freshly-hatched and isolated miracidia
at a concentration of 200 nM dsRNA in CBSS plus glucose. Controls included larvae cultured
in CBSS alone or CBSS containing a decoy sequence with similar GC content. Miracidia were
allowed to transform to sporocysts in the presence of siRNAs and further cultivated for
7 days, after which sporocysts were photographed with a digital camera attached to a
fluorescent inverted microscope and observed for various phenotypes including failure/
delay in transformation, loss of movement, tegumental lysis/death and size changes.
For the later phenotype, sporocysts lengths were measured from captured images using
Metamoph software and statistically analyzed. The relative amount of some known transspliced transcripts was measured by quantitative PCR (qPCR). Results: From the sequencing
of the cDNA libraries, a total of 2644 sequences were obtained, and after assembly, they
resulted in 603 unique sequences that were annotated. For silencing of trans-spliced
genes, a set of phenotypes were evaluated, but only larval size seemed to be affected by,
at least, one of the two SL siRNA treatments. Additionally, some transcripts identified by
the analysis of the SL enriched cDNA libraries, such as, Calcium Channel, ATPase Inhibitor,
Fosfoserin-Hidrolase, Thioredoxin and Enolase were tested by real-time qPCR for their
abundance and showed a significant level reduction (at least 50%) after siRNA treatment,
when compared to control and others non-trans-spliced transcripts. The search for
potential GO enriched terms in SL sequences will be performed in order to find biological
processes that are more represented among trans-spliced transcripts. Conclusions: RNAi
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results demonstrate the efficacy of siRNA treatment in suppressing the expression of transspliced transcript levels, although, we were unable to completely disrupt the trans-splicing
mechanism. The definable phenotypic changes in larval development could represent a
disruption of developmental signals, nutrient processing or metabolic imbalance.
375
IdEntIfICatIOn Of a COnSERvEd dOMaIn RICH In B CELL EPItOPES WItHIn SOLUBLE
atP dIPHOSPHOHYdROLaSE fROM SCHISTOSOMA MAnSOnI BY SYntHEtIC PEPtIdES
anaLYSES
Priscila de faria Pinto; ana Carolina R. g. Maia; Rita gabriela P. R. Mendes; Karen t. flores;
Cristiane Carvalho Campos; Michélia antônia n. gusmão; gabriane nascimento Porcino;
Michelle Lima detoni; Marcos Luiz de Oliveira Penido; Maria aparecida Juliano; Luiz Juliano;
Rodrigo Correa Oliveira; Paulo Marcos zech Coelho; Eveline gomes vasconcelos
Introduction: Evolutionary and closer structural relationships were demonstrated between
Solanum tuberosum apyrase and a soluble S. mansoni ATP diphosphohydrolase isoform by
phylogenetic analysis, alignment of the primary amino acid sequences, epitopes prediction
and by hypothetical three-dimensional models. Specific protein domains were suggested to
be potentially involved in the immune response, and also seem to be conserved during host
and parasite co-evolution (Parasitology 135:943, 2008). Cross-immunore¬activity between
potato apyrase and IgG antibody from S. mansoni-infected mice or schistosomiasis pa¬tients
strongly suggested that the epitopes shared between the vegetable and parasite proteins
are antigenic (Mem. Inst. Oswaldo Cruz 105:370-379, 2010). These results prompted us
to analyze a specific conserved domain using synthetic peptides. Methodology: Peptides
belonging to the conserved domain from both S. mansoni ATP diphosphohydrolase (Sm)
and its potato apyrase (pot) counterpart were designed based on a strategic comparative
study, which maintained predicted epitopes available for antibody and HLA-DR binding.
The peptides were obtained by solid-phase synthesis, and their molecular mass and
purity were confirmed by amino acid analysis and by MALDI-TOF. The serum samples of
patients with schistosomiasis (n= 35), seropositive for potato apyrase, were selected from
a collection of samples from endemic areas for S. mansoni that are studied by the Instituto
de Pesquisas René Rachou (CPqRR). These patients (age range 4-83 years; 20 male and 15
female) were diagnosed by positive quantitative parasitological stool examinations by the
Kato-Katz method, and had a low intensity of infection (182 ± 50 eggs per gram of faeces).
Patients co-infected with other parasites were excluded from this study. Sera (n= 12) from
healthy individuals from an area non-endemic for schistosomiasis were used as controls.
The total IgG, IgG1 and IgG4 antibody reactivities against synthetic peptides (10 μg/well)
were evaluated by ELISA, using serum samples diluted 1:50 (IgG1 and IgG4) and 1:100 (total
IgG), peroxidase-conjugated anti-IgG, anti-IgG1 or anti-IgG4 human-specific immunoglobulin
antibodies and OPD/H2O2 as the substrate, and the results were expressed in optical density.
Immunostimulatory property of each peptide was also tested. Results: The IgG antibody level
against SmB1LJ (C, 0.155 ± 0.046; T, 0.231 ± 0.046; cutoff 0.247; 8/35, 23% seropositivity),
SmB2LJ (C, 0.199 ± 0.052; T, 0.305 ± 0.099; cutoff 0.378; 12/35, 34% seropositivity), SmB1MP
(C, 0.275 ± 0.052; T, 0.448 ± 0.099; cutoff 0.303; 30/35, 86% seropositivity), potB1MP (C,
0.209 ± 0.042; T, 0.295 ± 0.111; cutoff 0.293; 19/35, 54% seropositivity) or potB2LJ (C, 0.331
± 0.060; T, 0.473 ± 0.118; cutoff 0.451; 19/35, 54% seropositivity) was significantly (P < 0.001)
higher than that found in healthy individuals (C). The IgG1 subtype level against SmB1LJ (C,
0.017 ± 0.019; T, 0.072 ± 0.060; cutoff 0.057; 20/35, 57% seropositivity), SmB2LJ (C, 0.015 ±
0.015; T, 0.050 ± 0.069; cutoff 0.045; 11/35, 31% seropositivity), potB1LJ (C, 0.003 ± 0.004; T,
376
0.022 ± 0.033; cutoff 0.011; 20/35, 57% seropositivity), potB1MP (C, 0.025 ± 0.008; T, 0.055
± 0.037; cutoff 0.041; 20/30, 67% seropositivity) or potB2LJ (C, 0.021 ± 0.017; T, 0.056 ±
0.038; cutoff 0.055; 16/35, 46% seropositivity) was significantly (P < 0.01) higher than that
found in healthy individuals (C). The IgG4 subtype level against SmB1LJ (C, 0.081 ± 0.033; T,
0.153 ± 0.047; cutoff 0.148; 19/35, 54% seropositivity), SmB2LJ (C, 0.109 ± 0.019; T, 0.154 ±
0.075; cutoff 0.148; 17/35, 49% seropositivity), SmB1MP (C, 0.043 ± 0.026; T, 0.113 ± 0.080;
cutoff 0.097; 19/30, 63% seropositivity), potB1LJ (C, 0.117 ± 0.026; T, 0.322 ± 0.198; cutoff
0.169; 25/35, 71% seropositivity), potB1MP (C, 0.083 ± 0.006; T, 0.102 ± 0.069; cutoff 0.097;
12/30, 40% seropositivity) or potB2LJ (C, 0.109 ± 0.018; T, 0.183 ± 0.067; cutoff 0.146; 25/35,
71% seropositivity) was significantly (P < 0.001) higher than that found in healthy individuals
(C). Comparative individual analysis demonstrated variable antibody reactivity. Inoculation
of each peptide in healthy Swiss mice had significant immunostimulatory ac¬tivity, increasing
the amount of total IgG antibody, and IgG1 and/or IgG2a subtypes, as observed by Dot blots
and ELISA. Conclusions: The significant seropositivity of total IgG antibody, and IgG1 or
IgG4 subtypes, demonstrated the high sensitivity of these peptides for antibody detection.
The immunostimulatory activities of the peptides in healthy mice confirm the epitopes
antigenicity. These results are in accordance with the existence of shared antigenic epitopes
between potato apyrase and soluble ATP diphosphohydrolase isoform, and demonstrated
that in schistosomiasis this domain from parasite protein is rich in B-cell epitopes. These new
biomolecules could be useful as a composition to improve diagnosis methods or vaccine
protocols in schistosomiasis studies. Financial Support: FAPEMIG, CNPq, UFJF.
377
IMMUnOStIMULatORY PROPERtIES Of a SYntHEtIC PEPtIdE BELOngIng tO tHE
SOLUBLE atP dIPHOSPHOHYdROLaSE ISOfORM fROM SCHISTOSOMA MAnSOnI, and
IdEntIfICatIOn Of tHIS PROtEIn In SOLUBLE Egg (SEa) and WORM (SWaP) antIgEnS
PREPaRatIOnS
Michélia antônia do nascimento gusmão; Michelle de Lima detoni; Rita gabriela Pedrosa
Ribeiro Mendes; Karen takaki flores; ana Carolina Ribeiro gomes Maia; gabriane nascimento
Porcino; Cristiane de Carvalho Campos; nayara Braga Emídio; Maria aparecida Juliano; Luiz
Juliano; Henrique Leonel Lenzi; Paulo Marcos zech Coelho; Priscila de faria Pinto; Eveline
gomes vasconcelos
Introduction: Previously a soluble ATP diphosphohydrolase was isolated from S. mansoni egg
and adult worm tegument. This parasite protein has cross-immunoreactivity with potato
apyrase, suggesting that they share conserved epitopes. By in silico analysis, a closer structural
relationship was demonstrated between potato apyrase and S. mansoni SmATPDase 2,
this last protein with predicted molecular mass 63.8 kDa, and with one amino-terminal
transmembrane region possibly subject to a proteolytic posttranslational processing that
results in a secreted protein of 55 kDa. Comparative studies of synthetic peptides belonging
to the hypothetical antigenic conserved-domains from this soluble ATP diphosphohydrolase
isoform and their potato apyrase counterparts facilitated the identification of a domain in
parasite protein containing epitopes reactive with IgG1 and IgG4 antibody from patients
with schistosomiasis. In this work, the immunostimulatory properties of a peptide (B2LJ)
that belongs to a conserved domain from this parasite protein were analysed. In addition,
the sera immune identified the soluble ATP diphosphohydrolase isoform in soluble egg (SEA)
and worm (SWAP) antigens preparations. Methodology: Polyclonal antiserum against B2LJ
was obtained from seven-week old Swiss mice that were inoculated by a peritoneal route
with two injections of B2LJ (10 μg); the first injection was emulsified in Freund’s complete
adjuvant and the other injection was in Freund’s incomplete adjuvant and delivered in 15day interval. The reactivity of IgG1 and IgG2a antibodies from pre-immune serum (control)
or immune serum bound to ELISA plates coated with B2LJ (10 μg/well) was quantified using
different serum dilutions (1:50-1:800) and peroxidase-conjugated isotype-specific secondary
antibody and OPD/H2O2 as substrate. For dot blots, purified potato apyrase (1 μg) or peptide
B2LJ (5 μg) was spotted onto nitrocellulose membranes, which were blocked and incubated 5
h with pre-immune sera (1:100), immune sera anti-B2LJ (1:100) or immune sera anti-potato
apyrase (dil. 1:1000). Total protein (100 μg) of SWAP or SEA was electrophoresed in 10% SDSPAGE, electroblotted onto nitrocellulose membrane, and the Western blots were developed
with either pre-immune serum or immune serum anti-B2LJ diluted 1:800. As positive control,
the Western blots were also developed with rabbit polyclonal serum anti-potato apyrase
(diluted 1:1000). The membranes were revealed by chemiluminescence with the specific
secondary antibody coupled to horseradish peroxidase and Luminol as substrate using ECL
kit and exposed to X-ray film. In addition, cryostat sections of infected mouse liver were
obtained 8 weeks post-infection with S. mansoni and submitted to immunocytochemical
techniques, using sera immune anti-B2LJ and confocal fluorescence microscopy. Results:
Dot blots of either potato apyrase or B2LJ were positive for rabbit polyclonal anti-potato
378
apyrase and mouse polyclonal anti-B2LJ antibodies, demonstrating that inoculation of
Swiss mice with B2LJ induced significantly higher levels of B2LJ-specific IgG antibody than
the control, and confirming the presence of homologue epitopes within vegetable protein.
The IgG1 (> 1:800) and IgG2a antibody (> 1:800) levels quantified by ELISA, using B2LJ as
coating antigen, were elevated, about 5-fold higher than those found in pre-immune serum.
In Western blots of both SEA and SWAP preparations, mouse polyclonal anti-B2LJ antibodies
recognized bands of approximately 63 and 55 kDa, the last possibly as a consequence of
a proteolysis of the 63 kDa band. Rabbit polyclonal anti-potato apyrase antibodies, used
as control in this experiment, recognized only the band of approximately 63 kDa in both
SEA and SWAP preparations. Pre-immune serum diluted 1:800 did not react with SEA or
SWAP. By confocal fluorescence microscopy, the S. mansoni egg ATP diphosphohydrolase
isoform was detected in von Lichtenberg’s envelope, and in the outer side of the egg-shell,
confirming that the soluble isoform is secreted. Conclusions: Inoculation of the B2LJ in
healthy Swiss mice has remarkable stimulatory activity, increasing significantly the amount
of IgG1 and IgG2a subtypes, which further support the antigenicity of the correspondent
domain within parasite protein during schistosomiasis progression. In addition, these results
suggest the presence of distinct epitopes in this peptide capable of inducing a Th1 or Th2type immune response. The anti-B2LJ antibody reactivity against SEA or SWAP confirmed
the identity of a soluble ATP diphosphohydrolase isoform in these antigenic preparations, as
well as the sensitivity and specificity of this immune serum. The peptide B2LJ could be useful
as a composition to improve diagnosis method or vaccine formulations. Financial Support:
FAPEMIG, CNPq, UFJF.
379
InfLUEnCE Of HEPatOSPLEnIC SCHIStOSOMIaSIS ManSOnI On tHE CaStELLI’S
IndExES.
adenor almeida Pimenta filho; Luiz arthur Calheiros Leite; Bianka Santana dos Santos; Caique
Silqueira Martins da fonseca; ana Lucia Coutinho domingues; vera Lucia de Menezes Lima
Introduction: Schistosomiasis is a parasitic disease that affects 200 million people worldwide.
Research using infected animals has shown changes in lipid metabolism, but there is
controversial about atherogenic effects. The ratios of Total Cholesterol/HDL-cholesterol (TC/
HDL-c), and LDL-cholesterol/HDL-cholesterol (LDL-c/HDL-c), have been used as predictor
factors for development of cardiovascular diseases. These ratios were called Castelli Index I,
and Castelli Index II, respectively. The purpose of the present study was to analyse the lipid
profile and Castelli Indexes in compensated hepatosplenic schistosomiasis (HS). Methodology:
101 individuals agreed to participate in this study (63 healthy individuals composed the
control group and 38 HS patients from Service of Gastroenterology of Clinical Hospital of the
Federal University of Pernambuco, Brazil). The lipid profile was determined by enzymaticcolorimetric methods, except VLDL-cholesterol and LDL-cholesterol that were calculated
by Friedewald‘s equation. Castelli’s Indexes I and II were also calculated. The statistical
differences between groups were accessed by unpaired “t” Test (p < 0.05), and data were
expressed as mean and standard error of the mean. Results: TC (149.6±4.6 vs. 176.7±4.6
mg/dL; p=0.0016), and LDL-c (85.5±4.3 vs. 117.4±4.9 mg/dL; p<0.0001) plasma levels were
significantly lower in HS group when compared to the control group. It was not found a
significant statistical difference about plasma levels of HDL-c, VLDL-c, and Triglycerides. The
Castelli’s Indexes I (3.6±0.2 vs. 4.3±0.2; p=0.0081) and II (2.1±0.2 vs. 2.9±0.1; p = 0.0007)
were lower in HS patients. Conclusions: These results suggest that the alterations in lipid
profile observed in compensated hepatosplenic schistosomiasis carriers may provide some
type protection against cardiovascular disease. However, other studies are necessary to a
better understanding about lipid metabolism in the chronic schistosomiasis. Supported by
CNPq, CAPES, and FACEPE.
380
InvEStIgatIOn Of HUMan Cd59 ORtHOLOgS In tHE SCHISTOSOMA MAnSOnI
gEnOME
Cibele aparecida tararam; Leonardo Paiva farias; Bogar Omar araujo Montoya; Henrique
Krambeck Rofatto; Willian Castro-Borges; Sophie Manoel; alan Wilson; Luciana Cezar de
Cerqueira Leite
Introduction: Schistosoma mansoni is the predominant parasite responsible for
schistosomiasis, which affects more than 200 million people worldwide, with a further 600
million individuals living at risk of infection in endemic areas. The treatment of the disease is
based on the drug praziquantel, but chemotherapy cannot prevent re-infection and there is
evidence supporting the development of drug resistant parasites, emphasizing the need for
a complementary approach. Therefore, the vaccination of susceptible individuals using key
S. mansoni antigens may be the most effective means of controlling schistosomiasis. With
the completion of the S. mansoni EST Project, it was possible to visualize a large repository of
schistosome genes, bringing insights into several aspects of schistosome biology in relation
to parasite survival and host-parasite interactions. These data resulted in the identification
of new potential vaccine candidates as the human CD59 orthologs. These genes belong to
the family of the antigen 6 of lymphocytes (Ly6), whose main member is the CD59 protein,
an important inhibitor of the membrane attack complex (MAC) by complement system.
Therefore, the objective of this work is the investigation of the human CD59 orthologs in
the S. mansoni genome and the characterization of one member of this family, Ly-6.5 gene;
besides the presentation of some results on Ly-6.1. Methodology: The alignment of the
CD59 orthologs was performed using the software Clustal X. Phylogenetic analysis was also
performed using these sequences and comparing with CD59 orthologs from S. japonicum, S.
hematobium, Fasciola hepatica and Homo sapiens. The presence of peptides, transmembrane
domains, GPI anchor and glycosylation sites was evaluated by Expasy tools. The mRNA
expression levels of the Ly-6 genes in the egg, miracidium, cercaria, schistosomulum and
adult stages were evaluated by real time RT-PCR and compared with microarray data. One
member of the family, Ly-6.5, was cloned into an expression vector and transformed into
Escherichia coli. The recombinant protein was obtained by purification in Ni+2 – Sepharose
affinity chromatography. Polyclonal antibodies were obtained by immunization of rats using
the recombinant protein and adjuvant. Western blot was performed to characterize the
protein in the life cycle stages. Immunolocalization assays using adult worms sections and
whole cercaria and schistosomulum were performed to investigate the localization of the
protein on the parasite. Molecular shaving assays were performed incubating adult worms
with the PiPLC enzyme that cleaves membrane-binding proteins by GPI anchor and analyzing
the samples by Western blot. Results: In S. mansoni, the Ly6 family is composed by six genes
(Ly6.1, Ly6.2, Ly6.3, Ly6.4, Ly6.5, Ly6.6) with 25-30% of identity with human CD59. They contain
Upar/Ly-6 domains, signal peptides, transmembrane domains, GPI anchors and glycosylation
sites. Most of them show increased gene expression in the schistosomulum stage by real
time RT-PCR, except Ly6.3 gene, which shows increased expression in eggs and Ly6.4 which
is increased in adult worms. These results were corroborated by microarray analysis. One
of the genes, Ly6.5, was cloned and expressed, and Western blot analysis revealed high
381
expression levels of the Ly6.5 protein in schistosomula; some in adult worms and cercaria;
little in miracidia and none in eggs. The Ly6.1 protein showed the same profile of expression
with the exception of miracidia, where the protein is not expressed and in male where the
expression is higher, as compared to female. The protein Ly6.5 was immunolocalized to
the tegument of 3h, 7 and 21day-old fixed schistosomula and live in vitro schistosomula by
confocal microscopy. The protein was not localized in cercariae, while adult sections showed
staining on the surface and inside, in the parasite body. Localization of the Ly6.1 protein was
analyzed only in adult worms and was verified on their surface. The characteristic of these
proteins to be membrane binding by GPI-anchor was confirmed for Ly6.1 and Ly6.5 by the
molecular shaving technique. Conclusions: The genes of the Ly6 family show similarity with
human CD59, which inhibits complement; most of them are up-regulated in schistosomulum
stage as determined by real time RT-PCR; and the proteins are associated with tegument.
These results indicate that family should be investigated as potential vaccine candidates.
Functional assays, the characterization of other members of the family and CD spectrometry
are underway.
382
InvEStIgatIOn Of vEnOM aLLERgEn LIKE PROtEInS (vaLS) fROM SCHIStOSOMa
MAnSOnI aS vaCCInE CandIdatES
Leonardo Paiva farias; Henrique Krambeck Roffato; Cibele aparecida tararam; Bogar Omar
Montoya; Rafaela Sachetto fernandes; Patricia aoki Miyasato; Eliana nakano; Iain W.
Chalmers; Samirah Perally; Paul Hensbergen; Ron Hokke; Karl f. Hoffmann; Luciana Cezar de
Cerqueira Leite
Introduction: Mining the database generated from S. mansoni transcriptome using Gene
Ontology categorization, potentially surface-exposed or exported proteins were identified.
Among the antigens tested as DNA vaccines, a gene with similarity to Venom-Allergen-Like
proteins (SmVAL5) conferred a worm burden reduction of 31 %. Further searches for similar
domains in the S. mansoni genomic data bank revealed 29 paralogs from this new gene family.
The SmVALs are members of a protein superfamily containing a conserved SCP/TAPS (Spermcoating protein/ Tpx-1/Ag5/PR-1/Sc7) domain, which may be important in host-pathogen
interactions. Based on the transcriptome profile, proteomics and microarrys data available
for these molecules we selected four members (SmVALs 4, 5, 7 and 26) to be investigated as
vaccine candidates. Methodology: Native or codon optimized versions of the selected SmVALs
genes were expressed in P. pastoris, and the recombinant proteins purified by nickel affinity
chromatography. To evaluate the protective potential of this molecules C57BL/6 mice were
inoculated subcutaneously (sc) with 3 doses of 25 µg of rSmVALs formulated with Freund’s
adjuvant. Six weeks after challenge infection, worms were collected and counted from the
hepatic portal vein. The immune response profile was characterized by ELISA and ELISPOT.
Additionally, proteomics and nano LC-MS/MS technologies were exploited to examine the
cross reactivity properties of the VAL family in S. mansoni using antibodies raised against
recombinant SmVAL4, -5 and -26. Results: Preliminary immunization and challenge assays
revealed that rSmVAL5 induced a 40% worm burden reduction, confirming the previous
protective potential for this molecule. The humoral immune response was characterized by
high levels of IgG1 and low levels of IgG2 antibodies; the cytokine profile revealed low levels
of IFN-g and high levels of IL-4; suggesting a more Th2 driven immune response. The cross
reactivity data revealed antibody cross-reactivity within phylogenetically-related SmVALs.
Furthermore, anti-SmVAL5 seems to have a broad binding capacity to different SmVALs
recognizing the SmVALs 5/15, 26/28, 27, 9 and 29, while the anti-SmVAL4 recognized SmVAL4,
18 and 19 and anti-SmVAL26 antibody cross reacted just with SmVAL 26/28. Conclusions: Ours
results suggest some potential for these molecules as vaccine candidates, but also highlights
that cross reactivity effects between different SmVALs expressed in different stages should
be considered in the design of a schistosomiasis vaccine. Financial Support: FAPESP, CNPq,
Fundação Butantan.
383
LaRgE-SCaLE gEnE ExPRESSIOn EvaLUatIOn Of SCHISTOSOMA MAnSOnI adULt
WORMS tREatEd WItH Mg132
Enyara Rezende Morais; Lizandra guidi Magalhães; Érika Bueno de Carvalho Moreira; Cláudia
Sossai Soares; Katia Cristina Pereira Oliveira; vanderlei Rodrigues
Faculdade de Medicina de Ribeirão Preto-USP, SP - Brasil
Introduction: Schistosomiasis remains a serious public health problem in many developed
countries. Countless reports show the increasing emerging of Schistosoma mansoni strains
Praziquantel-resistent, choice drug for schistosomiasis treatment. In this manner, it is
necessary the development of new drugs that are able to act against this parasitic disease. The
effect of proteasome inhibitors, as MG132, has been described in some protozoan parasites,
inhibiting the growth or cycle progression, or blocking the replication. Our laboratory results
showed that MG132 have reduced the number of pulmonary schistosomula, parasitic burden
and therefore, the oviposition in infected mice. In this regard and from the knowledge
of that this compound reduces directly the proteasome activity and can modulate the
expression of some genes, we propose to investigate the effects of MG132 in S. mansoni
adult worms through microarray analysis. Thus, the aim of the current study was to better
understand the ubiquitin-proteasome signaling pathway in this parasite biology, as well as
to analyse new alternative molecular targets in the therapeutics against the schistosomiasis.
Methodology: Adult worms were obtained from portal vein perfusion of mice 56 days after
infection and separated in two groups with 20 couples each one. The first, control group
was maintained in RPMI culture medium with 10% DMSO, since MG132 was dissolved in
DMSO. In the second group, 50 μM MG-132 was added into culture medium. Both groups
were maintained at 37ºC, 5% CO2, during 24 hours. The colorimetric quantification assay,
based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), was used
to check the parasites viability. The total RNA from adult worms, treated or untreated
with MG132, was extracted with Trizol reagent, using the recommended protocol. After
Trizol extraction, RNAs were treated with Qiagen DNAse I and subsequently purified using
Qiagen RNAeasy mini kit. The integrity of RNA samples was evaluated using microfluidic
electrophoresis in the Bioanalyzer equipment. The gene expression analysis was performed
using the 44K oligoarray, an oligonucleotide microarray slide with around 44.000 probes
representing Schistosoma mansoni genes, idealized by Verjovski-Almeida et al. (2007). Cy5and Cy3-labeled cRNA was obtained using 500 ng of total RNA from four independent pools
of MG132-treated or control adult worms as templates for amplification of poly-A RNA by
T7-RNA polymerase with the Agilent Quick Amp Labeling Kit. The purified labeled cRNA
was used in the microarray hibridization. The slides were washed and processed according
to Two-Color Microarray-Based Gene Expression Analysis (Quick Amp Labeling) Protocol
and scanned on a GenePix 4000B scanner. Data were extracted using Feature Extraction
software, low intensity data points were filtered out according to this software criteria. Total
intensity data from each experiment were normalized by Quantiles Normalization Method,
excluding positive and negative external controls. The Significance Analysis of Microarray
(SAM) was used as the statistical test to identify differentially expressed genes and these
were considered as significantly differentially expressed at q-value ≤ 0.025. Hierarchical
clustering of selected genes was generated using Spotfire Decision Site software. Results:
384
The preliminar analysis showed a set of 3520 probes identified with a statistically significant
(q-value ≤ 0.025) differential expression between MG132 treated and control adult worms.
More analyses will be developed to identify the genes represented by these probes and
each differentially expressed gene will be classified according to the biologic process, cellular
component and molecular function using the Gene Ontology (GO) annotation. Conclusions:
So, the gene expression analysis will be useful to evaluate the drugs effect, as MG132, in the
parasite, aiming to amplify the knowledge about the S. mansoni biology. In this case, it will be
able to provide new drug targets and that can be important for the new drugs and vaccines
construction for schistosomiasis prevention and control.
385
LOOKIng fOR BIOMaRKERS Of SCHIStOSOMIaSIS In a SIMPLIfIEd SERUM PROtEOME
Jonatan Marques Campos; Leandro xavier neves; Milton Hércules guerra de andrade; Renata
guerra de Sá; vanderlei Rodrigues; William de Castro Borges
Introduction: Following the extensive characterization of the S. mansoni proteome, the
application of functional proteomics in experimental models of infection, may constitute an
interesting approach towards understanding the tissue-associated complications resulting
from parasite homing in the hepatic portal system. In parallel, the identification of parasite
molecules present in body fluids of the vertebrate host, especially blood and urine, may
provide novel diagnostic methods for schistosomiasis. This would result in overcoming the
cross-reactivity associated to helminth antigens when antibody-based assays are employed.
Nevertheless, the detection of biomarkers in the serum constitutes a biochemical challenge.
Highly abundant proteins such as albumin, immunoglobulins, complement factors and
protease inhibitors compromise the identification of the low abundant components in the
serum, masking potential biomarkers of the disease. Our present strategy aims to employ
serum depletion methods using two separated techniques: protein precipitation by organic
solvent and affinity chromatography. These two approaches will exclude the highly abundant
serum components facilitating the identification of proteins present in response to a S.
mansoni infection. Methodology: Blood from control and S. mansoni infected animals, were
obtained by cardiac puncture and allowed to coagulate at room temperature. Serum samples
were obtained by centrifugation at 1500 x g. Pools from infected and control animals were
diluted 1:3 in ultra pure water, following addition of acetonitrile to a final concentration
of 60%. Samples were then sonicated for 10 min in a ultrasonic water bath, vortexed and
further sonicated for 10 min. Precipitated proteins from each sample were recovered by
centrifugation at 12.000 x g for 10 min at 4°C and resuspended in 0.01M phosphate buffer
pH 7.2, containing 50 mM NaCl. The supernatants were vaccum-concentrated to dryness and
the obtained pellet solubilized in the same buffer. Protein samples recovered from control
and infected animals were analysed by 1-DE (15% SDS-PAGE) after Coomassie staining of
the gel. Prior desalting using acetone precipitation allowed a comparative 2-DE analysis of
the acetonitrile ‘resistant’ fractions to be performed. For isoelectric focusing, proteins were
separated in a 7 cm pH 3-10 IPG strip, followed by electrophoresis in a 15% SDS-PAGE, for the
second dimension. Results: Analysis of the acetonitrile ‘resistant’ fractions from both control
and infected animals revealed the predominance of low molecular mass proteins ranging
from 10 to 75 kDa, indicating depletion of the most abundant serum components. It was
possible to observe quantitative and qualitative differences in the band patterning obtained
from these two groups, which were better detected using the 2-DE technique. Acetonitrile
depleted proteins analysed under 1-DE also revealed pronounced differences associated to
the serum of infected animals. Conclusions: Over 90% depletion of the serum proteome has
been achieved with precipitation using organic solvent. This technique will permit detailed
inspection of the low-mass range protein components present in the serum of S. mansoni
infected animals. Mass spectrometry analysis of the observed serum protein alterations is
currently under way to uncover possible biomarkers of the disease.
386
MaCOPHagE-dERIvEd HEdgEHOg LIgandS PROMOtES fIBROgEnIC RESPOnSES In
SCHIStOSOMIaSIS ManSOnI
thiago de almeida Pereira; Wing-Kin Syn; Izabela voieta; Jiuyi Lu; Steve S. Choi; Kolade M
agboola; gamze f Karaca; Carlos M antunes; William E. Secor; zilton a andrade; José R.
Lambertucci; fausto Edmundo Lima Pereira; anna Mae diehl
Universidade Federal do Espírito Santo, ES - Brasil
Introduction: Schistosomiasis affects over 200 million people worldwide. In hepatic
schistosomiasis mansoni, eggs deposited in the mesenteric veins are carried into the
microvasculature of the liver and induce a granulomatous reaction that evolves to portal
fibrosis. The mechanisms driving schistosomal fibrosis are not fully elucidated. Macrophages
are one of the major components of the granulomas and are capable of inducing myofibroblast
activation and collagen production. Since in other chronic liver diseases the Hedgehog
(Hh) pathway was shown to play a pivotal role in hepatic stellate cell activation (HSC) and
fibrosis progression, our aims were to determine if Hh pathway activation occurs during
fibrosis progression in chronic schistosomiasis mansoni and to determine if macrophagerelated mechanisms are involved. Methodology: Immunohistochemistry (IHC) was used to
evaluate Hh pathway activity in 28 wedge liver biopsies from patients with different grades
of schistosomiasis fibrosis staged by ultrasound (WHO criteria; pattern A=3, D=5, Dc=2,
Ec=17,F=1). A macrophage cell line (RAW 267.4) and primary rat Kupffer cells (prKC) were
stimulated with 10ug of soluble egg antigen (SEA) or vehicle for 6 hours and then RNA
was isolated for analysis by qRT-PCR. Conditioned media from prKC (+/- SEA) were used to
stimulate Shh-LightII cells and firefly and Renilla luciferase activities were determined using a
dual luciferase kit after 48 hours of incubation. To verify that induction of luciferase was due
to Hh signaling initiated by macrophage-derived Hh ligands, reporter cells were treated with
macrophage-conditioned media ± Cyclopamine or its inactive analog Tomatadine. Results:
Patients with schistosomiasis expressed more Hh ligands (Shh and Ihh) and target genes
(Patched and Gli-2) than healthy individuals; fibrous septa were particularly enriched with
Hh pathway positive cells. Liver ductular and stromal cells produced and responded to Hh
ligands. Double IHC for Ihh and CD68, a marker of monocytes and macrophages, showed that
Ihh positive cells in the fibrous septa were also CD68 positive. In vitro studies showed that SEA
stimulated cultured RAW 264.7 and prKC to express Ihh and Shh mRNA (p<0.05). Conditioned
media from the SEA-treated macrophages/KC induced luciferase production by Shh-LightII
cells significantly more than conditioned medium from vehicle-treated macrophages/KC
(p<0.001) and Cyclopamine inhibited this effect (p<0.001). Conclusions: Schistosomal egg
antigens stimulate liver macrophages to produce Hh ligands that activate Hh signaling in
Hh-responsive cells. This may promote hepatic fibrogenesis during schistosomiasis mansoni
infection.
387
MInIng tHE SCHISTOSOMA MAnSOnI SOLUBLE PROtEOME fOR Ig-LIKE MOLECULES
Leandro xavier neves; Karina taciana Santos Silva; Jonatan Marques Campos; Milton Hércules
guerra de andrade; Renata guerra de Sá; vanderlei Rodrigues; William de Castro Borges
Introduction: A common fold consisting of a beta-sandwich formed of 7 strands in 2 sheets
with a Greek-key topology defines the Immunoglobulin domain. This domain is present in a
number of proteins with diverse functions such as antibodies, cell adhesion molecules and
tyrosine kinase receptors. Bioinformatic analyses using the SchistoDB revealed the presence
of the Ig fold in distinct molecules of parasite origin involved in cell signaling, adhesion and
tegument composition. In the present work we employed an affinity purification strategy
for the capture of immunoglobulins from human and rat sera, which also proved useful for
binding Ig-like molecules in the soluble extract of adult S. mansoni parasites. This approach
is likely to provide enrichment for relevant effector molecules with the potential to be used
in proteomic investigations. Methodology: Ten grams of CL-4B sepharose beads were first
activated using epichloridrine, aminated, followed by succinylation and further activation
using N-hydroxysuccinimide. The activated beads were then allowed to react with 500
mg of compound 22* (C22), Sigma, for 1 h at room temperature. Affinity columns were
mounted onto 3 mL syringes for subsequent protein purification. Human and rat sera were
diluted 1:10 in distilled water and loaded separately onto C22-sepharose columns. Unbound
material was washed out of the columns using distilled water. Bound material was eluted in
0.01 M phosphate buffer pH 7.2 containing 50 mM NaCl, and 1 mL aliquots were recovered.
Protein elution was monitored using a spectrophotometer at 280 nm. Aliquots from peak
fractions were separated under denaturing conditions on a 12% SDS-PAGE, followed by
Coomassie staining. Tryptic digestion of the two major bands enriched by the C22-sepharose
columns was performed and peptides subjected to analysis in a LC-IT-TOF (Shimadzu) mass
spectrometer. Mass spectral data were submitted to NCBInr database searching using
Mascot. The identity of the eluted proteins was confirmed by the Western blotting technique.
Approximately 10 μg of the purified material from rat serum was separated on a 12% SDSPAGE and the gel transferred to a PVDF membrane. After a blocking step in TBST / 5% milk,
anti-rat IgG alkaline phosphatase labeled antibody was added to a final dilution of 1:1000,
and incubated for 2h at room temperature. The membrane was developed using the alkaline
phosphatase substrates NBT (Nitro-Blue Tetrazolium Chloride) and BCIP (5-Bromo-4-Chloro3‘-Indolyphosphate p-Toluidine). C22-sepharose column was finally tested for its ability to
bind surrogate molecules in a soluble extract of adult S. mansoni parasites. Approximately
300 mg of parasites (wet weight) were homogenized by sonication (5 series of 30 pulses) in
2 mL of 25 mM Tris-HCl pH 7.5, 1 mM DTT supplemented with 1 x Protease Inhibitor Cocktail
(Sigma). The soluble protein content was recovered by centrifugation at 20.000 x g for 30
min. C22-sepharose affinity chromatography using the soluble worm protein preparation was
performed as described above and the bound material eluted in 0.01M phosphate buffer pH
7.2 analysed by 12% SDS-PAGE, followed by Coomassie staining of the gel. Results: Analysis
of the C22-sepharose bound material by 1-DE revealed the presence of two major bands
of approximately 25 and 50 kDa. In gel digestion of the respective bands followed by mass
spectrometry analysis allowed the identification of the Ig light and heavy chains. The same
388
protein bands were detected by the anti-rat IgG (whole molecule) antibody, confirming the
capture of immunoglobulins from serum. A similar affinity purification experiment, employing
soluble extract from adult parasites, demonstrated that C22-sepharose enriches for at least
four defined bands under Coomassie staining. Identification of these Ig-like molecules
from parasite origin is currently under way. Conclusions: We have successfully produced
a novel affinity column able to bind immunoglobulin from serum. In addition, supported
by the existence of Ig-like molecules encoded by the S. mansoni genome we showed that
our affinity strategy can be useful to enrich for selected molecules from a soluble worm
preparation. Mass spectrometry analysis will reveal if C22-sepharose chromatography will
become an approach to mine the S. mansoni proteome for Ig-like molecules. * Unrevealed
due to data protection.
389
MOLECULaR MOdELIng StUdIES On nOvEL InHIBItORS Of CatHEPSIn d-LIKE
EnzYMES fROM SCHISTOSOMA MAnSOnI
ana Carolina Rennó Sodero; Suzanna Romero de Sousa neves; Salvatore giovanni de Simone;
floriano Paes Silva-Jr
Introduction: Peptidases located in the gut of schistosomes parasites digest host blood proteins
as a nutritional source and have been described as potential targets for chemotherapy.
In 1997, Wong and co-workers identified an aspartyl protease from Schistosoma mansoni
similar to human cathepsin D (here denoted as SmCD1). However, two novel cathepsin D-like
proteases of S. mansoni were identified by our group and denominated as SmCD2 and SmCD3,
indicating that a family of aspartyl proteases is present in this parasite. The objective of this
work was to study the structural aspects of the inhibition of SmCD proteases by pepstatin
and the statin-derived peptidomimetic inhibitors LBPP1, LBPP37 and LBPP38 in order to
rationalize experimental results for the inhibition of the aspartyl protease activity contained
in an aqueous extract of adult schistosome worms (hereafter called SmAP). Homology models
were generated for the enzymes SmCD1 and SmCD2, since only their sequences are available,
and the docking methodology was performed to generate enzyme-inhibitor complexes.
The results of the molecular modeling studies have been used to discuss the molecular
determinants for SmCD1 and SmCD2 selectivity and to propose strategies for the rational
design of improved inhibitors of SmCD proteases. Methodology: The sequence alignments
of the enzymes with templates were obtained via T-Coffee server and the molecular models
were built by Modeller 9v4 program. Suitability of the generated structures was inspected
using the PROCHECK, WhatCheck, Verify-3D and Prove computer programs. Docking studies
of pepstatin and the three peptidomimetic inhibitors were performed utilizing the ligand
binding data obtained from kinetic experiments. The initial structures of the pepstatin and
the three peptide inhibitors were built on the basis of the structure of CH-66 inhibitor from
mouse submaxillary renin complex (PDB code 1SMR). AutoDock 4.2 was used for all docking
calculations. An initial population of 300 randomly placed individuals, a maximum number
of 5,000,000 energy evaluations and 150 runs were performed by using Lamarck genetic
algorithm searches. A mutation rate of 0.1 and a crossover rate of 0.6 were used. Results
differing by less than 2.0 Å in positional root-meansquare deviation (RMSD) were clustered
together and represented by the result with the most favorable free energy of binding. The
complexes were analyzed by visual inspection and automated assignment of interatomic
contacts using CSU/LPC server. Results: The analysis of the Ramachandran plot for the two
models indicated that more than 90% of the residues were located in the favored regions and
less than 1% had disallowed conformation. Other stereochemical parameters were inside
the expected values. The models show 3D structures similar to other aspartic proteases,
since folding is highly conserved in the A family of MEROPS database. Three disulfide
bonds were detected within the SmCD1 model, which are highly preserved at the aspartic
proteases family (C46-C53, C210-C214 e C253-C290). On the other hand, only two disulfide
bonds were identified within the SmCD2 model (C46-C53, C239-C276). The 3D coordinates of
SmCD1 and SmCD2 models were utilized in the docking methodology to predict the structure
of the inhibitors complexes. Considering the average predicted binding energy within the
390
top scoring cluster for each enzyme-inhibitor complex, LBPP37 and LBPP38 showed higher
affinity than LBPP1. This is probably because the change in conformation of Gln side chain
of LBPP37 and 38 (due to imposition of ciclohexil and benzyl substituents, respectively)
suggesting that those ligands interact through hydrogen bonds with Gly221. This residue is
described in literature as critical for all aspartic protease function and the absence of this
interaction can help to explain the low score of LBPP1 by Autodock. In addition, residue
Tyr78 made hydrophobic interactions with P1 substituent of ligands. Phe115, Phe120 and
Ile123 characterized a hydrophobic pocket within the S1 subsite, producing significant
interactions with all inhibitors. Pepstatin and LBPP-1, the most active compounds, produced
an extensive hydrophobic interaction with Ile13 and Ile 31 in SmCD2 and Val31 in SmCD1.
The least active compound (LBPP37) does not interact with such residues probably due to
gauche conformation of ciclohexyl substituent. Besides, LBPP1, LBPP37 and LBPP38 made
non-attractive contacts with Ala13 and Ile131 in SmCD1, and Ile13 in SmCD2. Conclusions:
The novel peptidomimetic inhibitors of aspartyl proteases of S. mansoni were docked into
SmCD1 and SmCD2 models. The Autodock scoring function were in good agreement with
experimental activities and allowed a structure-based rationalization of kinetic data. Finally,
our future work is to improve this analysis by molecular dynamics simulations.
391
MOLECULaR, IMMUnOLOgICaL and MORPHOLOgICaL anaLYSIS Of SCHISTOSOMA
MAnSOnI SCHIStOSOMIaSIS COMBInEd tHERaPY WItH aRtEMISInIn-PRazIQUantEL.
ana Júlia Pinto fonseca Sieuve afonso; Silvana Belo; Isabel Clemente; Ilda Jeremias; Claudia
Pen; Cátia alexandra Costa ferreira; Maria amélia grácio
Universidade Nova de Lisboa, Instituto de Higiene e Medicina Tropical, Unidade de
Helmintologia e Malacologia Médica, Unidade de Parasitologia e Microbiologia Médica Portugal
Introduction: Schistosomiasis is caused by digenetic trematode of the genus Schistosoma.
For the treatment of this disease Praziquantel (PZQ) is mostly the drug of choice. However,
its extensive use in mass treatment for schistosomiasis control potentiates the appearance of
resistance or increased tolerance to the drug. Therefore, the use of combination therapy may
be an alternative to improve cure rates thus eventually delaying the permanent establishment
of drug resistance. One of the drugs that has been suggested to be used in combination
with PZQ is Artemisinin (ART). Methodology: In this study we used the rodent model of
Schistosoma mansoni and divided into three groups: 1 - mice infected with S. mansoni which
were administered 40 mg/kg/dose of PZQ, the standard curative dose in humans; group 2 mice infected with S. mansoni treated with 30 mg/kg of ART during five days; group 3 - mice
infected with S. mansoni treated with a combined therapy of PZQ (40 mg/kg/dose) and ART
(30 mg/kg) during five days. Negative and positive controls were included. On three time
points during these experiments serum and blood for DNA extraction were collected (on the
day of drug administration, on the 30th and 45th days post drug administration). On the 45th
day mice were sacrificed and the spleen, the liver and the intestine were removed for imunohistological analysis. Liver and blood were also collected for RNA extraction. The RNA was
used for cDNA synthesis and Real Time PCR analysis of genes that might be involved in drug
metabolization. Results: Immunological, histological and genetical analyses were performed.
Real time PCR techniques were use to evaluate differential expression of genes involved in
drug metabolism. All the three experimental groups presented a very significant reduction
on the total parasite load on day 15th post drug administration, but only experimental group
2 (ART administration) presented a total reduction that was maintained until day 30 post
drug administration. This might be an indication that S. mansoni strain we are using in the
laboratory presents a significant reduction in its sensitivity to PZQ. On day 45 post drug
administration it was still experimental group 2 (ART) that presented the biggest reduction
on parasite load, when compared to the group that was given PZQ (group 1) and PZQ plus ART
(group 3). Conclusions: Anti-schistosome IgG and IgM levels presented significant differences
in the three experimental groups suggesting that different immunological responses from
the host is dependent upon the drug administered. Again the group where that difference
was more significant was experimental group 2 where ART was administrated. Together with
the information on the parasite load reduction observed in this group this might suggests
that ART efficacy should be further analysed as a potential schistosomicide drug, not only
as a single drug therapy regimen but also as a combination therapy with PZQ for example.
Keywords: Schistosomiasis, drug resistance, Praziquantel, Artemisinin.
392
MORPHOMEtRY and gEnEtIC anaLYzES Of BulInuS GlOBOSuS (gaStROPOda
PLanORBIdaE) fROM REgIOn Of BEngO, angOLa
Ângela Helena C. velez; Maria Manuela P. Calado; ana Júlia afonso; Maria amélia a. grácio
Instituto de Higiene e Medicina Tropical - Brasil
Introduction: Bulinus globosus, a freshwater snail, plays an important role as intermediate
host of Schistosoma haematobium, the etiological agent of human urinary schistosomiasis.
With a geographic distribution throughout the coastal plain of Angola and in the northern
highlands, B. globosus can be found in diverse habitats, ranging from large lakes to small
ponds, irrigation canals and small waterways. So, in Angola, this mollusk is considered as
an important responsible for the transmission of S.haematobium (Carvalho et al, 1960 and
Grácio, 1980). This study aimed to evaluate the genetic diversity of two populations of B.
globosus in the region of Bengo (Angola), located north of the capital, Luanda, with the use
of PCR-RFLP technique applied to the ITS region of ribosomal RNA (rRNA). Methodology: The
snails were collected from two ponds - Ibêndua and Sungue, situated in the Bengo region
- between February and April 2009. For morphometric study were measured 311 shells of
B. globosus and recorded their total height (th), height of the aperture (ha), maxim width
(mw) and width of the aperture (wa), and the relationships between these dimensions were
determined. The calculation of maximum and minimum, average, median and standard
deviation was based on the methodology of Anderson et al (1961). Statistical analysis was
carried out using SPSS version. 17.0., and the Kruskal-Wallis test to compare the median
of the various morphometric parameters. For genomic DNA extraction and molecular
analysis were randomly selected 45 snails. The genomic DNA was performed according to
the protocol CTAB / chloroform (Stothard & Rollinson, 1997). In amplification of genomic
DNA were used oligonucleotide ETTS1 and ETTS2 (Kane and Rollinson, 1994). PCR products
were visualized in agarose gel 1% (w/v). An enzymatic digestion of PCR products which
used the following restriction enzymes: Msp1, RsaI, and AluI. The analysis of the different
polymorphism - RFLP was performed as described by Raahange and Kristensen (2000). The
remaining PCR product was purified and sequenced. The sequences were aligned with the
support of the program MEGA 3.1, and compared with homologous sequences available
in GenBankdatabase. Results: In the analysis of morphometric parameters of the shell
results showed inter-population differences but were not statistically significant (p> 0.05).
The amplification of the ITS region of the samples studied showed fragments approximately
1200 bp. After enzymatic digestion these fragments presented polymorphic bands ranging
from 700pb to 100bp depending on the enzyme used, thus this allowed us to observe intraspecific genetic polymorphisms for the different habitats included in this study. Conclusions:
The results, though preliminaries seem to agree with other authors (Stothard et al 1996),
indicating that the PCR-RFLP technique can be applied to the genotyping of Bulinus globosus.
As for the sequences obtained, even partial, allowed to establish a homology between our
sequences and those deposited in the database.
393
PaRtIaL CHaRaCtERIzatIOn Of BIOMPHAlARIA TEnAGOPHIlA (taIM StRaIn)
tRanSCRIPtOME USIng tHE ExPRESSEd SEQUEnCE tag StRatEgY – PRELIMInaRY
RESULtS
Michele araújo Pereira; aristeu Silva-neto; Marina de Moraes Mourão; thomaz Lücher-dias;
Consuelo Latorre fortes-dias; Paulo Marcos zech Coelho; glória Regina franco
Introduction: Snails of the genus Biomphalaria are intermediary hosts for the trematoda
Schistosoma mansoni. B. tenagophila is the second most prevalent species in Brazil, having a
great epidemiological importance for schistosomiasis transmission, especially in the south of
the country. Populations of such species are either highly susceptible or resistant to infection
with different strains of S. mansoni. Identification of genes and biochemical pathways involved
in the immune mechanisms of Biomphalaria resistance against S. mansoni infection could
reveal valuable information and to develop approaches to control and prevent this parasitic
disease. Aiming for a better understanding of the mechanisms of resistance and interaction
between the snail and S. mansoni, we focus our transcriptome study in B. tenagophila from
the Ecological Reservoir of Taim-RS, a lineage resistant to different strains of S. mansoni.
Methodology: Total RNA was isolated from the cephalopodal region of B. tenagophila Taim
by the Trizol method and the mRNA was obtained by chromatography in oligo(dT) column.
cDNA library was constructed to generate Expressed Sequence Tags (ESTs) using the Kit
CreatorTM SmartTM cDNA Library Construction (Clontech). cDNA molecules were produced,
amplified and then size-selected by gel. The size-selected cDNAs were cloned into the pDNRLIB vector. Bacteria were transformed and PCR reactions were performed to evaluate the size
of the inserts and to select clones for sequencing. Plasmids were extracted from the bacterial
clones and sequenced on the automated DNA Sequencer MegaBace 1000. Similarity
searches against DNA and protein non-redundant databases, using the Basic Local Alignment
Search Tool (BLAST) programs, were done as an attempt to identify genes expressed by B.
tenagophila. Results: From the 1440 clones randomly selected by size, 144 were sequenced.
From these, some sequences represented the 12S and 18S subunits of the mitochondrion
rRNA from B. tenagophila Taim and two sequences showed high similarity to B. glabrata and
B. pfeifferi. The remaining sequences produced low quality data or represented unwanted
regions of the vector or poly(A) tail. Conclusions: The cDNA library has not shown the
expected quality. Therefore, we decided to start a new cDNA library construction, changing
some experimental conditions to avoid these biases and achieve a higher quality library.
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PROtEOMIC anaLYSIS Of SCHISTOSOMA MAnSOnI tREatEd In vItRO WItH 3MEtHYLCLOnazEPaM
Jean Pierre Barros thibaut; Regina Coeli gonçalves Lage; vagner Simonin; antonio galina;
guilherme Oliveira; françois noel
Introduction: Clonazepam (CLO) and 3-methylclonazepam (MeCLO) induce contraction
of adult male Schistosoma mansoni in vitro and have a proven schistosomicidal activity in
humans. In an attempt to clarify the mechanism of this effect, we decided to use a proteomic
approach and to map the changes in protein expression of S. mansoni worms treated in
vitro with MeCLO. Methodology: Adult worms were obtained by perfusion of portal and
mesenteric veins of infected mice and distributed in tissue culture dishes containing RPMI.
The worms were kept at 37°C in an atmosphere of 5% CO2 in the air for 24 hours before
addition of 3μM MeCLO. After 24h of this treatment, the worms were examined under a
Nikon Eclipse TC-100 inverted microscope for ensuring that they were alive and then treated
for 1 h with a lysis buffer (urea 8M, thiourea 2M, chaps 4%, DTT 50mM, protease inhibitor
and H2O) followed by mechanical digestion using an insulin syringe. The homogenate was
thereafter centrifuged at 16000 g. The protein concentration was determined by the Bradford
method (1976). The pellets were submitted to two-dimensional electrophoresis (2DE) and
the 2DE gel images were analyzed using the PDQuest program. The protein expressed
exclusively in the treated worms, or whose expression was at least 2,5-fold higher than in
the control worms, were identified by MALDI-ToF-ToF mass spectrometry using the database
of predicted proteins from the S. mansoni genome and the MASCOT program. In another set
of preliminary experiments, we determined the activity of phosphoglycerate kinase (PGK),
fructose 1,6 biphosphate aldolase (FBA) and glyceraldehyde 3-phosphate dehydrogenase
(G3PDH) present in S. mansoni homogenates indirectly by measuring the consumption
of NADH at 340 nM. For the PGK activity, the samples were evaluated in a Tris-HCl buffer
100mM pH7.5 with, NADH 0.3mM, MgCl2 2mM, ATP 1mM, EDTA 1mM and glyceraldehyde
3-phosphate dehydrogenase 5u.mL-1 and 3-phosphoglicerate 5mM as substrate. For the FBA
activity, the samples were evaluated /assayed in Tris-HCl buffer 100mM pH7.5 with, NADH
0.3mM, Triosephosphate isomerase 10u.mL-1, glyceraldehyde 3-phosphate ehydrogenase
1u.mL-1 and fructose 1,6 bisphosphate 5mM as substrate. For the G3PH activity, the samples
were evaluated / assayed in a Tris-HCl buffer 50mM pH7.5 with NADH 0.3mM, MgCl2 2mM,
ATP 1mM, EDTA 1mM and phosphoglycerate kinase 13u.mL-1 and 3-phosphoglicerate 5mM
as substrates. Results: The proteomic analysis revealed 9 exclusive spots and 4 spots with
enhanced expression in the treated group. After spectrometral identification, these spots
were classified in two major groups of proteins: structural proteins like troponin T, actin-1
and tubulin beta chain possibly related to the immediate effects of MeCLO such as spastic
contractions followed by morphological changes. The other group of proteins, such as PGK,
FBA and G3PDH are enzymes participating in the glycolysis known to be the archetype of
an universal metabolic pathway occurring with variations, in nearly all organisms, either
aerobic or anaerobic. In preliminary experiments, we were able to measure the activity of
these enzymes in untreated S. mansoni with the following activities: 0.6963, 0.3349 and
0.4727 nmol NAD/mg protein for PGK, FBA and G3PDH respectively. These results indicate
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that it would be feasible to do such measurements in treated worms in order to verify if
the increased expression of the 3 glycolityc enzymes is accompanied by an increase in their
enzymatic activity. Conclusions: Proteomic analysis of S. mansoni maintained in culture and
treated with MeCLO revealed increases in the expression of proteins related to the worm’s
musculature and morphology and in enzymes participating in glycolysis, one of the major
metabolic pathways of the parasite. This latter original observation could indicate that
MeCLO inhibits the aerobic metabolism of the worm leading to overexpression of enzymes of
the glycolitic pathway in an attempt to overcome the metabolic collapse. Such experimental
approach may aid to better understand the parasite’s biology and the schistosomicidal
effects of MeCLO.
396
PROtEOMIC and HIStOPatHOLOgICaL anaLYSIS Of PRazInQUantEL tREatMEnt On
LIvER Of MICE InfECtEd WItH SCHISTOSOMA MAnSOnI
gabriela ayres fragoso nascimento; Humberto gonçalves Bertão; adriana Silva andrade
Pereira; natália Lima Cavancanti; Maria da Paz Carvalho da Silva; Mário Ribeiro de Melo
Júnior; Mônica Camelo Pessoa de azevedo albuquerque; Maria Elizabeth Cavalcanti Chaves
Introduction: Human schistosomiasis is caused by trematode flatworms Schistosoma, being
considered one of the neglected tropical diseases in the world. The infection caused by this
helminth results in several metabolic disfunctions in host organism, mainly resulting in hepatic
fibrosis. This fibrosis may be represented by small focal areas of chronic inflammation and
excess extracellular matrix deposited around the periovular granulomas, which are distributed
in variable numbers at the periphery of the portal vein system. Currently, praziquantel (PZQ)
is one the drug of choice in the treatment of three major species of schistosomes that infect
the humans (S. mansoni, S. haematobium and S. japonicum). In this work, proteomic and
histopathological analysis of liver from mice infected with S. mansoni and treated with PZQ
were done in order to observe the changes in protein expression due to the treatment with
the drug. Methodology: Male mice with 6 weeks old were infected with 50 cercaria of the
BH (Belo Horizonte) strain of S. mansoni. Sixty days post-infection, the animals were divided
in six groups of 10 animals each. Each group received a daily dose of 12.5 mg of PZQ by
orogastric gavage except the control group and were sacrificed on consecutive days until
the end of 120hrs [T0(0hs), T1(24h), T2(48h), T3(72h), T4(96h) and T5(120h)]. Animals were
anesthetized with 46.4mg of ketamine hydrochloride plus 4.6mg of xylazine hydrochloride.
The livers were washed with saline, one of the lobes frozen at 80o C to proteomic studies
and the other sectioned, fixed in formalin and processed for histopathological evaluations.
Briefly, all samples were embedded in paraffin, sliced and stained for the following methods
Hematoxylin-eosin (HE), van Giemson, Periodic Acid Schiff (PAS) and Masson Trichrome. For
the 2D-electrophoresis 200µg of protein from of the total liver crude extracts were solubilized
in rehydration solution containing 9M urea, 0.5% (w/v) CHAPS, 0.2% (w/v) DTT, 0.5% (v/v)
IPG buffer (pH 3-10) and 0.002% of bromophenol blue. The isoelectric focusing (IEF) were
done as recommended by the manufacture (GE Healthcare, USA), followed by a 10% SDSPAGE. The analysis of the 2D gel was made through the digitization of the images and the
attributes (pI and molecular weight) of protein spots determined by use of the software
ImageMaster 2D Platinum (GE Healthcare, USA). Results: 2D-electrophoresis was used to
measure the biological variation in protein abundance from livers of animals treated and
untreated with PZQ after infection with S. mansoni. It was observed a diminished amount of
proteins in T5 (108 spots) comparing with the groups T0 (112 spots). There was difference
between the proteins expressed in the groups, after matching treated and untreated gels, it
was observed that 63 spots were present in both gels, 45 only in the treated group and 49 in
the untreated group. It is important to study the difference of protein profile found, to one
possible explication of the effect caused in the liver after using the drug. The histopathological
study revealed that most groups shows nonspecific inflammatory process of character
perivascular interstitial with lympho-plasmocytary and eosinophilic infiltrate, especially in
groups T3(72h), T4(96h) and T5(120h), with rare presence of multinucleated giant cells,
397
despite the formation of granulomatous response periovular. Fybroplasia was observed in
all groups and was more intense collagen deposition in the liver parenchyma in the T1(24h)
and T5(120h). The results of counts of granulomas in liver of mice in experimental groups
showed a random distribution according to the time of treatment. The groups T2(48h) and
T5(120h) had a mean count above 10, while groups T0(0h), T1(24h), T3(72h) and T4(96h) had
a count less than 10 granulomas. In each group, the average score showed that there wasn´t
a pattern of the reduction of the granulomas with increasing treatment. Conclusions: The
pharmacoproteomic is important for the identification of specific therapeutical targets for S.
mansoni. Although the number of spots in the treated and untreated gels has been almost
the same, the protein profile differed in the assayed groups. These proteins may be targets
of future studies for the identification of their structures and metabolic effects in the host
organisms. The histopathological study did not present a pattern of decrease in the number
of granulomas with the increase of treatment, however showed a reduction in the number
of eggs and inflammatory cells in the hepatic tissue.
398
PURIfICatIOn Of tHE EnzYME URIdInE CYtIdInE KInaSE Of SCHISTOSOMA MAnSOnI
SELECtEd aS a tHERaPEUtIC taRgEt
débora Meira neris; Larissa Romanello; alexandre Cassago; Humberto d‘Muniz Pereira;
Richard Charles garratt; fernanda de freitas anibal
Introduction: According to the World Health Organization (WHO), in tropical and subtropical
areas, schistosomiasis is the second most prevalent parasitic disease in terms of socioeconomic and public health, being surpassed only by malaria. Caused by a trematode of
the genus Schistosoma, it is still a disease with great potential to spread, with more than
200 million people infected in 76 countries. In the infected host, the disease is characterized
by hepatic granuloma and tissue fibrosis. Thus, the parasite-host relationship can lead to
morbidity and result in hepatosplenomegaly, hepatic fibrosis and ascites. The granulomatous
process in schistosomiasis is dependent on CD4 + T cells and requires recruitment and
accumulation of inflammatory cells at the site of the eggs deposition. Granulomas are
composed primarily of macrophages, eosinophils and lymphocytes. The pathogenesis of
schistosomiasis depends on a number of factors: the parasite strain, age, nutritional status
and immunity of the host and, especially, the parasite load. The emergence of resistant strains
for currently used antischistosomal drugs indicate the necessity to develop more efficient
drugs with fewer side effects or even a vaccine. For that, to study new treatment options, in
which the investigation of enzymes present in the critical pathways of S. mansoni becomes a
path of interesting research. Studies of new treatment options based on the investigation of
enzymes in the metabolic pathways of nucleotides of Schistosoma mansoni becomes a path
of interesting research. In this context, studying the effects of the enzyme Uridine Cytidine
kinase (UCK) S. mansoni in the recovery of worms and eggs in relation to parasite load to the
effects of animals infected with S. mansoni, may hint at new options for therapeutic targets.
One of the essential enzymes for the metabolism of S. mansoni enzyme Uridine-cytidine
kinase, a pyrimidine ribonucleosides kinase that catalyzes the phosphorylation of uridine or
cytidine to UMP and CMP respectively. The genome of S. mansoni has two isoforms of the
UCK Smp_062260.1 (UCK1) and Smp_062260.2 (UCK2) coding for proteins with 245 and 264
amino acids respectively. The two isoforms share 97% sequence identity deferring only the
C-terminal, where Smp_062260.2 has 19 residues more. The two isoforms share 46% identity
when compared to human UCK. Methodology: The two isoforms of Uridine-cytidine kinase of
S. mansoni have been expressed satisfactorily and conditions for expression of the UCK1 was
37°C and 100 micromolar of IPTG for 3 hours, to UCK2 the temperature was 15°C and 100
micromolar of IPTG overnight. The yield was 30mg/L to UCK1 and 8mg/L to UCK2. Results: After
determining the optimal conditions of expression the purification of affinity chromatography
was made. The vector pET28a and Petsumer inserts in the N-terminal protein a sequence
of six histidines, this tail polihistidina has an affinity for zinc or cobalt ions. The experiments
were used to purify the resin chromatography of nickel or cobalt and the analysis was made
in poliacrilamina gel. Conclusions: Our results showed that the yield of protein expression
occurs at significant levels, and the UCK1 had a higher yield in shorter time compared to UCK2.
Yet, from the purification, further studies with the enzyme may provide interesting tools to
further develop related therapy, since it participates in pathways essences of the parasite
399
RECOMBInant SMnPP-5 IndUCES antIBOdIES tHat PaRtIaLLY InHIBIt tHE SURfaCE
EnzYMatIC aCtIvItY BUt faIL tO PROtECt agaInSt CHaLLEngE WItH SCHISTOSOMA
MAnSOnI
Henrique Krambeck Rofatto; Leonardo Paiva farias; Cibele aparecida tararam; Bogar Omar
araujo Montoya; Robert alan Wilson; Luciana Cezar de Cerqueira Leite
Introduction: Schistosomiasis affects 200 million individuals in several countries, including
Brazil; its treatment is based on praziquantel, but chemotherapy does not prevent reinfection and does not reverse the pathology induced by the eggs, emphasizing the pursuit for
a more effective approach. Recent proteomic characterization of the S. mansoni tegument,
the major parasite-host interface, identified a putative nucleotide pyrophosphatase/
phosphosdiesterase (SmNPP) 5, as a plasma membrane-associated protein. NPPs are
ubiquitous membrane-associated or secreted ecto-enzymes that act by regulating the
metabolism of extracellular nucleotides, consequently having a role in purinergic signaling,
which affects diverse biological processes such as platelet aggregation, apoptosis, cell
proliferation, differentiation and motility. In the current work we characterized the protein
at the host-parasite interface and evaluated the potential of this protein as a vaccine
candidate. Methodology: The gene was cloned by RT-PCR from adult S. mansoni RNA,
heterologous expression was obtained in E. coli and the protein was purified by nickel affinity
chromatography. The recombinant protein was used to produce polyclonal antibodies, which
were employed to characterize the profile of protein expression throughout the life cycle,
to localize it in adult worm sections and to evaluate if they were capable to inhibit the
enzyme activity of ex vivo live adult worms. The recombinant protein was also evaluated
in immunization and challenge assays; mice were subcutaneously immunized with the
recombinant protein formulated with Freund’s adjuvant and then challenged with cercariae.
After forty five days the animals were perfused to measure the worm burden; the immunized
mice immune response profile before and after challenge was characterized by ELISA and
ELISPOT to measure the levels of antibodies isotypes and cytokines, respectively. Results:
The protein was expressed by E. coli as inclusion bodies, it was solubilized with 8M urea
and purified under denatured conditions by nickel affinity chromatography; the refolding
was performed by slow dialysis, but the protein precipitated. The protein expression profile
throughout the life cycle was determined by immunoblotting; demonstrating low levels of
expression in schistosomula and high levels in adult worms, not being detected in the other
stages; it was also verified that the protein is more expressed in males than in females and
is enriched in the adult tegument fraction. The surface-exposed localization of the protein
at parasite-host interface was confirmed by immunolocalization and the protein was not
recognized in other tissues of adult worms sections. Antibodies produced by immunization
with the recombinant protein were able to partially inhibit (~60%) the enzymatic activity in
ex vivo live adult worms. Despite of induction of a specific immune response, no reduction in
worm burden was observed in the immunized animals; the humoral immune response was
characterized by high levels of anti-SmNPP-5 IgG1 and low levels of IgG2 antibodies before
and after the cercarial challenge and the cytokine profile revealed low levels of IFN-g and high
levels of IL-10 and IL-5; suggesting a more Th2 drifted immune response. Conclusions: The
400
SmNPP-5 is a protein present in the host-parasite interface, probably involved in important
adaptive and survival processes in the blood vessels. Although immunized animals developed
a specific immune response, it was not enough to kill the worms and it is possible that it will
be necessary to obtain the recombinant protein in its native form to generate a stronger
specific immune response capable to completely inhibit the worm enzyme and therefore
obtain better protective levels. To achieve this goal we will attempt expression of this protein
in an eukaryotic expression system, Pichia pastoris. Different routes of immunization with
different adjuvants will be evaluated as well. Financial support: FAPESP.
401
REdUCtIOn On COnCEntRatIOn Of LECItHIn-CHOLEStEROL aCYLtRanSfERaSE In
PLaSMa Of HEPatOSPLEnIC SCHIStOSOMIaSIS PatIEntS
Maciel, g.R.; Carvalho, v.C.O.; Silva, C.a; Santos, B.S. dos; domingues, a.L.C; Lima, v.L.M.
Introduction: Schistosomiasis mansoni is an infectious and parasitic disease that affects around
200 millions of people in the world. Around 10% of the patients present the most severe form
of the disease – the Hepatosplenic (HS) form. Individuals with HS present abnormalities in the
lipid and lipoprotein metabolism. Several studies have shown decrease of Lecithin-Cholesterol
Acyltransferase activity (LCAT, EC 2.3.1.43) in patients with schistosomiasis mansoni, which
can be due to reduced LCAT mass or abnormality on its substrates. The aim of this study was
to evaluate the influence of HS on LCAT concentrations. Methodology: Concentration of LCAT,
measured by using an ELISA method (DAIICHI, Japan), was evaluated in plasma of 33 patients
with HS proceeding from the “Service of Gastroenterology of Clinical Hospital of the Federal
University of Pernambuco, Brazil”, and in plasma of 30 healthy individuals, of both genders.
The differences in LCAT concentrations amongst the groups were accessed through unpaired
“t” Test (p < 0.05). Results: LCAT concentration in plasma of HS patients was reduced by 33%
in men and 40% in women, when compared to the respective control group. Conclusions:
The results suggest that the decrease found in LCAT activity in HS patients may be due to an
reduction in the synthesis of LCAT by the liver, or a consequence of an abnormal removal of
LCAT from the circulation.
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REfInEMEnt StRUCtURE adEnOSInE KInaSE and ExPRESSIOn HYPOxantHInEgUanInE PHOSPHORIBOSILtRanfERaSE Of SCHIStOSOMa ManSOnI
Larissa Romanello; alexandre Cassago; Ricardo de Marco; glaucius Oliva; Richard Charles
garratt; Humberto d´Muniz Pereira
Instituto de Física de São Carlos (IFSC), SP - Brasil
Introduction: Schistosoma mansoni is a human parasite causing schistosomiasis, which is
one of the most prevalent infectious diseases in the world: around 200 million people have
been infected. This parasite does not posses the “the novo” purine pathway and depends
on it’s host for purine requirements and therefore has been proposed as a potential target
for antischistosomal chemotherapy. The enzymes Adenosine Kinase (AK) (E.C2.7.1.20) and
HGPRT (EC2.4.2.8) are key components of this pathway. AK catalyzes reaction: adenine +
ATP <-> AMP +ADP and HGPRT: IMP + diphosphate <-> hypoxanthine + 5-phospho-alphaD-ribose 1-diphosphate. The main objectives of this project is the cloning, expression,
crystallization and structure resolution of the enzymes Adenosine kinase and Hypoxanthineguanine phosphoribosiltranferase from Schistosoma mansoni. Methodology: The cDNA of
the AK1, AK2 e HGPRT was amplified from a enriched cDNA library prepared by RT-PCR, total
mRNA, using specific reverse primer by PCR. The gene was digested with restriction enzymes
and was inserted in the pET28a expression vector digested with the same enzymes. The
recombinant plasmid was used in the transformation of E. coli BL21 (DE3). We tested various
conditions to expression, the best was 37ºC (AK2), 18ºC (HGPRT) and 100μM of IPTG. HGPRT
purified using affinity column cobalt yielding 10mg/L, concentrated until 6mg/mL and was
submitted to robotic crystallization trials. AK2 purified using two affinity steps (nickel and
AMP-agarose) yielding 4mg/L, concentrated until ~4mg/mL and was submitted to robotic
crystallization trials. Results: The AK2 was crystallized in 100mM Bis-tris pH6.5, 25% PEG 3350
and 200mM Li2SO4. The structure of the complexes AK-adenosine-AMP and AK-adenosine
were obtained in the LNLS MX2 beamline up to 2.3Å and 2.25Å. The crystals of AK2 belongs
to the space group P21212 with cell dimensions of a=59.9Å, b=180.3Å and c=78.3Å (AKAde-AMP) and a=55.74Å,b=168.69Å and c=75.04Å (AK-Ade). The AK-Ade-AMP complex was
solved by molecular replacement (MR) using Phaser employing human AK as a search model
(1BX4) and refinement was completed using the programs Coot and Phenix. The structure of
the AK-Ade was also solved by MR employing refined AK. Conclusions: Comparison of both
structures revels the movements in the protein with different natural ligands. We expected
that Schistosoma AK and HGPRT structures could help the development of new compounds
against schistosomiasis. Supported by Fapesp and CNPq.
403
SCHISTOSOMA MAnSOnI aLKaLInE PHOSPHataSE: dIffEREnCES In ExPRESSIOn
PattERnS aLOng LIfE CYCLE In tHE COntExt Of vaCCInE dEvELOPMEnt
Bogar Omar araujo Montoya; Cibele aparecida tararam; Henrique Krambeck Rofatto;
Leonardo Paiva farias; Luciana C.C. Leite; R. alan Wilson
Introduction: An effective vaccine against schistosomiasis remains an elusive goal for science.
Because of this reason, we focused our research in a molecule that has potential as vaccine,
the Schistosoma mansoni Alkaline Phosphatase, SmAP. Since many years ago, this enzyme
has been studied, nevertheless, little is known about its expression pattern along life
cycle stages. In this research, we intended to analyse its expression at both transcriptional
and translational level in eggs, miracidia, cercariae, 7-day old culture schistosomula and
separated sexes. Methodology: qRT-PCR amplification was performed in a GeneAmp®PCR
System 9600 (Corbett Research). For Western Blot, 20 µg of total protein from the stages
were electrophoresed and transferred onto PVDF membranes (Amersham Pharmacia Biotech
Limited, England). Blots were incubated with corresponding immunosorbed rat antisera and
specific antibody binding was visualized using the ECL Western Blotting Detection System
(Amersham Pharmacia Biotech). For immunolocalization, eight micrometer-cryostat sections
of adult worms were adhered to silanized glass slides (DakoCytomation) and incubations
were made with corresponding immunosorbed rat antisera. Specific antibody binding was
visualized with Alexa Fluor® 488 rabbit anti rat IgG (H+L) (Invitrogen) with a Zeiss LSM
510 Meta Confocal System. Activity assays were performed with the synthetic substrate
p-Nitrophenyl Phosphate (pNPP) on live worms. Results: We could determine very high
levels of transcripts in eggs and also in cercariae, and low levels in adults, miracidia and
7-day old culture schistosomula. Protein levels were high in eggs and adults, and low in
miracidia, cercariae and 7-day old schistosomula. There are increasing protein expression
levels in 3h- and 12h-old culture schistosomula. Immunolocalization experiments revealed
the presence of the protein in all the tissues in both male and female adult worms, with
a strong fluorescence especially in the vitelline glands of female adult worms. In addition
to this, activity assays of the native molecule in the live worms allowed seeing that it is
exposed in the tegument, with highest activity in females, and lowest activity at the 7 dayold schistosomulum stage. However, when evaluated the blocking activity of sera obtained
from rats immunized with the recombinant protein, it was not possible to see any inhibition
of activity in the native protein at the tegument surface with live worms. Conclusions: The
high protein levels in 3h- and 12h-old schistosomula and the high level of transcripts in
cercariae would point out the parasite’ strategy of having availability of transcripts ready to
be translated into protein as soon as it encounters the definitive host. SmAP is important in
adult female worms, especially in the production of eggs due to the strong fluorescence of
the vitelline glands. The low protein expression level seen in 7 day-old culture schistosomula
would explain why the previously reported anti-SmAP immune response observed in murine
schistosomiasis occurs only fifty days post-infection, when the schistosomes are in the adult
stage. Besides, the native protein is accesible to substrate molecules but even so, it cannot
be neutralized by any antibody raised against the recombinant inactive form, suggesting the
importance of generating neutralizing antibodies.
404
SCHISTOSOMA MAnSOnI gEnOMIC and fUnCtIOnaL gEnOMICS data IntEgRatIOn:
SCHIStOdB.nEt
guilherme C. Oliveira; Roney S. Coimbra; Jessica Kissinger; fabiano Mirsky Pais; Mariana C.
Simões; adhemar zerlotini
SchistoDB (www.SchistoDB.net) is a genomic database for Schistosoma mansoni. The database
uses the Genomics Unified Schema, GUS, and currently contains S. mansoni sequence data
(Genomic, EST) in a single user-friendly database. This release also contains ~13 thousand
automated gene predictions. SchistoDB offers a variety of tools including BLAST, protein motif
searches, keyword searches of pre-computed BLAST results, Gene Ontology assignments and
protein family information. In addition, we have mapped microarray probes. Recently data
from a number of transcriptomic experiments (microarray and SAGE) were also uploaded.
The combination of an annotated genome and a relational architecture has facilitated the
integration of the genome with other types of data and permitted the construction of
automated analysis pipelines. Integrated into SchistoDB are SchistoCyc and KEGG DRUG.
SchistoCyc is a prediction of the metabolic pathways of the organism based on the genomic
information. Genomic data were used to computationally predict metabolic pathways and
select reactions that might play a crucial role in parasite metabolism. A database called
SchistoCyc was created using SRI PathwayTools software which permitted us to predicted
112 metabolic pathways. KEGG DRUG contains chemical structure and data on drugs that
target orthologous proteins of other organisms. We have also linked gene products to the
TDR Targets database. SchistoDB allows the user to save and combine queries using Boolean
logic. The creation of this database has provided a platform for Schistosoma bioinformatics.
SchistoDB.net provides the community with a simple, user friendly interface that enables
powerful mining of the genomic data; and has been widely used. Future work will involve
the integration of additional data such as SNPs, deep seequencing transcriptomics and
genomics data, among others. Support: NIH (TW007012), SECTES/FAPEMIG (1181/08), CNPq
(306879/2009-3 and 573839/2008-5).
405
SCHISTOSOMA MAnSOnI vaCCInE CandIdatE antIgEn SCREEnIng USIng PROtEOMIC
tOOLS
fernanda Ludolf Ribeiro; Rosiane a. da Silva-Pereira; Paola Patrocínio; andréa gazzinelli
Corrêa de Oliveira; Rodrigo Corrêa Oliveira; guilherme Corrêa Oliveira
Introduction: The schistosomiasis control strategy is mainly based on the treatment of
infected individuals by chemotherapy with the Praziquantel. While the use of this drug
has an effect on morbidity, it does not prevent the re-infection, especially to those people
who live in endemic areas. The development of drug resistance by the parasite is also a
concern that has to be considered. Therefore, the development of a long term protection
based on vaccination is still necessary. The main objective of this work is to select novel
Schistosoma mansoni vaccine candidate antigens using serum from individuals living in
schistosomiasis endemic area. Methodology: Total proteins of S. mansoni adult worm and
schistosomula were submitted to bidimensional electrophoresis separation and blotted
onto PVDF membranes. To probe the bidimensional western blots, we used serum pooled
from infected or non-infected individuals from a schistosomiasis endemic area in Brazil.
Antigens recognized by these sera were identified by mass spectrometry. Results: The results
indicate that there was no correlation between the amount of adult worm proteins in a
bidimensional electrophoresis gel and their antigenicity pattern. Some weak Coomassie Blue
stained spots were strongly recognized by serum from infected individuals. In addition, there
were clear qualitative and quantitative differences in the profile of protein spots recognized
by the serum pools. Some of adult worm antigens were recognized only by the serum
from infected individuals. However, until now, none were reproducibly recognized only by
the serum from non-infected individuals living in endemic area. In order to evaluate if the
carbohydrate portion of glycoproteins was recognized by the serum pools, we analyzed the
immunoreactive profile of adult worm blotted proteins deglycosilated by periodate treatment
of PVDF membranes. The same spots blotted onto periodate treatment and not treated
membranes were recognized by both serum pools, indicating that the protein portion was
recognized by the sera. We also analyzed the immunoreactive profile of schistosomula total
proteins using the same serum pools. The results showed clear differences in the repertoire
of antigenic proteins of adult worm and schistosomula total protein extracts. Furthermore,
some immunoreactive spots from the adult worm total protein extract were collected to be
identified by mass spectrometry. Many of the proteins identified until now had already been
related as immunogenic protein. Others are yet uncharacterized proteins. Conclusions: Some
new S. mansoni antigenic proteins were identified in this project and we will test them as
potential vaccine candidates using recombinant proteins or DNA vaccines in murine model.
406
StRUCtURE Of MEtHYLtHIOadEnOSInE PHOSPHORYLaSE (MtaP) fROM SCHISTOSOMA
MAnSOnI
Juliana Roberta torini de Souza; alexandre Cassago; Ricardo de Marco; glaucius Oliva; Richard
Charles garratt; Humberto d‘Muniz Pereira
Instituto de Física de São Carlos, SP - Brasil
Introduction: The schistosomiasis is a chronic parasitic illness, caused by the parasite
Schistosoma mansoni that affects approximately 200 million people worldwide with
approximately 6 million in Brazil. Schistosomiasis is treated by the use of drugs that are
not-in fact effective for the eradication of the disease, and although their efficiency cause
serious side effects. The Schistosoma mansoni parasite does not posses the de novo pathway
for purine bases biosynthesis and depends entirely on salvage pathways for its purine
requirement. Thus this pathway can be identified as a potential target for the development
of specific drugs to combat schistosomiasis. The MTAP enzyme (EC 2.4.2.28) is key
component of this pathway, using 5’-deoxy-5’-methylthioadenosine (MTA) as it substrate.
MTAP catalyze reaction: S-methyl-5’-thioadenosine + phosphate = adenine + S-methyl-5thio-α-D-ribose 1-phosphate. Methodology: The MTAP amplification was performed using
a cDNA library from enriched mRNA of the adult worm. The cloning was done in pET28a
expression vector and confirmed by colony PCR. E. coli BL21-CodonPlus (DE3) was used
to express the SmMTAP, in 2XTY medium supplied with 50 μg/mL kanamycin and 10μg/
mL chloramphenicol. The expression was induced with 100μM/mL IPTG, the protein was
purified with affinity chromatography technique using Talon resin (Clonetech). The purified
protein was dialyzed with 20mM Tris pH 7.4, 200mM NaCl and 5mM β-mercaptoethanol and
submitted to a robotic crystallization trials using Honeybee 939 robot (Genomic Solutions).
Results: The MTAP was purified in large amounts with a high level of purity as monitored
by SDS PAGE. The yielding was 60mg per liter of 2XYT medium. The protein was crystallized
in 100mM Bis-tris pH 6.1-6.5 and 14-18% PEG 3350. Diffraction data was obtained in the
MX2 beamline of the LNLS up to 2.0Å. The crystals of MTAP belongs to the space group
P21 with cell dimensions of a=81.31Å, b=82.60Å, c=150.30Å and ɣ=100.9o. The MTAP-Ade
complex was solved by molecular replacement (MR) using Phaser employing human MTAP
as a search model (1BX4) and refinement has been carried using the programs Coot and
Phenix. The partially refined resulted a Rwork(%) = 20.5 and a Rfree(%) = 25.7. Conclusions:
The structures of the MTAP–ligand complexes provide a map of the active site and suggest
possible roles for specific residues in substrate binding and catalysis. This project belongs to
the Puri-pyrimidome Project that aims to obtain all the structures and kinetics constants of
all enzymes involved in nucleotide metabolism of S. mansoni.
407
tHE dEtECtIOn Of SCHIStOSOMa MAnSOnI TRAnSREnAl dna In URInE SaMPLES
USIng tHE MURInE MOdEL
guilherme Correia Oliveira; Martin J. Enk; nilton Barnabe Rodrigues
Introduction: Schistosomiasis caused by S. mansoni is a considerable public health problem
in countries of Latin America, the Caribbean and Africa ranking only behind malaria in the
list of important parasitic diseases. The diagnostic method of choice for the detection of this
disease is the Kato-Katz technique which identifies parasite eggs in stool samples. Besides
providing basic information about prevalence, it is one of the few techniques which quantify
the infection, giving estimates about the individual worm burden. Although this method
is relatively inexpensive and simple to execute, it presents a disadvantage, which is a lack
of sensitivity especially in low endemic areas and among individuals with low infection
intensity. Under these circumstances more sensitive techniques have to be developed. PCRbased diagnostic techniques relying on the detection of S. mansoni DNA in feces, serum,
plasma and urine have shown high sensitivity and specificity and offer an option to fill this
gap. In this work we present a S. mansoni trans-renal DNA (trDNA) PCR in murine model as an
alternative approach for the diagnosis of schistosomiasis. Methodology: In order to explore
the appearance of trDNA after infection 15 Swiss mice were infected with 25 cercariae each
and another 15 uninfected animals served as control group. Urine samples of each infected
animal were collected daily until the 10th day after infection. DNA from the urine samples was
extracted using a salting out and resin methodology and amplified using primers targeting
the 121 bp tandem repeat DNA sequence of S. mansoni, previously designed by Pontes.
Results: The urine examination according to the above described technique revealed one day
after infection a number of 6 positive among the 15 tested animals. Until the fifth day after
infection all 15 animals of this group were positive for schistosomiasis. Non of the uninfected
animals of the control group showed a positive result for schistosomiasis in the urine samples
collected at the same point of time as among the animals of the infected group. Conclusions:
The animal model using Swiss mice is a well known and valuable tool for the study of the
host-parasite relationship and they have been widely used to answer fundamental questions
on the dynamics of S. mansoni infections, including issues related to diagnosis. As the results
clearly show, PCR assay directed to trDNA in urine samples of mice is capable to detect
infection during a very early stage and low infection intensity. This alternative approach for
the diagnostics of S. mansoni infections applied in human urine samples has considerable
potential to detect infections during the prepatent period and individuals with low worm
burden. In this context further studies are necessary using the murine model, especially to
clarify time limits of the S. mansoni trDNA clearance after treatment.
408
tHE ROLE Of IROn In tHE MOdULatIOn Of gRanULOMatOUS RESPOnSE In MURInE
SCHIStOSOMIaSIS ManSOnI MOdEL
alice Maria de Magalhães Ornelas; flávia Rachel Moreira Lamarão; guilherme de Bustamante
Pereira de Miranda; Bernardo Miguel de Oliveira Pascarelli; Marcelo Ribeiro alves; Milton
Ozório Moraes; Marcelo Pelajo Machado
Introduction: Schistosomiasis is a chronic infectious disease that culminates in several
responses with intensities ranging from mild to death depending on its severity and/or
host susceptibility. Some organs like liver and intestine may be severely affected, showing
a granulomatous reaction to eggs released by adult helminths in circulation. Inside the
liver, fibrosis and loss of function can happen. The main point of this study is based on the
fact that iron has a central role in many physiological and physiopathological processes in
mammals and schistosomes. The increase or decrease of this element can change tissue
and cellular dynamics, leading to various consequences, such as oxidative stress (based
on the Fenton reaction) and subsequent cellular and tissue damage. For some diseases, it
has been already described a relationship between the local amount of iron and fibrosis,
including: hemochromatosis, hepatitis C, liver nonalcoholic fatty disease and liver alcoholic
disease. Here, we investigated the role of excess or scarcity of iron in the modulation of
the hepatic granulomatous reaction caused by Schistosoma mansoni in a murine model.
Methodology: First, five days old Swiss Webster albino mice were percutaneously infected
with 70 S. mansoni cercariae and divided in two groups. The first group received, by
intraperitoneal route (i.p.), a total dose of 100mg/Kg of ferrous sulfate (FS) distributed at
the 50th (50mg/Kg), 57th (25mg/Kg) and 64th (25mg/Kg) days of infection. The second
group received three i.p. doses of 15mg of desferrioxamine (DFO) each, an iron chelator,
at 50th, 51st and 52nd days of infection. A control group was established by giving 15ml
of water for injection (WFI) i.p. to infected animals at the same treatment points. Also,
non-infected animals treated the same way were also included in this experiment as other
controls. In all groups, the mice were killed when 90 days of infection were reached, for
the collection of liver and small intestine specimens, which were fixed in Carson`s Millonig
Formalin, embedded in paraffin, sectioned in 5 micrometer-thick slices and stained with
Hematoxylin-Eosin, Lennert’s Giemsa, Picrosirius Red and Perls for light microscopy analysis.
Other fragments were submitted to molecular biology protocols in order to determine the
expression of molecules related to homeostasis (hepcidin 1 and 2, transferrin receptor (TfR)
and IL-6), fibrosis (TGFb) and angiogenesis (VEGF). Results: The main results were: a) the
infected animals had higher survival when treated either by DFO or FS, b) less viability of
eggs were also observed with both treatments, c) there was marked intestinal retention
of eggs in the DFO and FS infected groups, sometimes causing vegetative lesions directed
to the peritoneal cavity (DFO), d) hepatic fibrosis was more intense in the infected group
treated with FS, e) some hepatic granulomas from DFO-treated animals were very loose;
sometimes the paracentral zone of these granulomas invaded the wall of neighbor portal
vein branches, establishing a communication with the intimal layer of these vessels; f) in
the same group, the extramedullary hematopoiesis observed in the outer layer of the many
hepatic granulomas appeared to be more pronounced and often associated with numerous
409
plasma cells; g) gene expression of most genes studied showed a great similarity between
DFO and SF infected groups. Conclusions: Altogether, our data confirm the interference of
positive and negative iron modulation on this experimental murine schistosomiasis mansoni
model progression and open new questions to be addressed in further approaches.
410
SCHIStOSOMIaSIS COntROL PROgRaM In aRaCaJU/SE: PROgRESS and PROBLEMS
nathalia vasconcelos Barroso, aloisio ferreira Pinto neto, Robergson Rozendo Ribeiro,
andréa valença Cardoso, Luciene Barbosa, Karina Conceição gomes Machado de araujo,
Satie Katagiri, Roseli La Corte dos Santos
Universidade Federal de Sergipe, SE,- Brasil
Introduction: The mainly goal of Schistosomiasis Control Program (PCE) is to reduce morbidity
from schistosomiasis through early diagnosis and treatment performed by active case
finding. Schistosomiasis Control Programs in Brazil began in 1976 with the Special Program
for the Control of Schistosomiasis (PECE). Specific treatment for Schistosomiasis has emerged
over the last 30 years, and the PCE have been remodeled according to the conjecture and
has contributed to the decline of prevalence rates and chronic forms, while the preventing
actions do not control the emergence of new foci, mainly in areas of urban sprawl. Objective:
To evaluate progress and problems in the PCE in Aracaju, using Santa Maria District as a model
standard. Methodology: A survey was carried out in to a database of the Center for Zoonosis
Control (CCZ) from Aracaju to identify the confirmed cases of schistosomiasis in the study area,
and then was initiated phase field with an active search of patients, and questionnaires and
delivery of collectors was distributed out for stools examination. Coprological examination was
carried out performing the quantitative Kato-Katz (one sample and 2 slides per person). Results:
In 2006, 12.075 examinations were performed, covering about 75% of the population from
the district. The analysis of stool samples showed, 688 people were infected by Schistosoma
mansoni. With the support of the Community Health Agents, responsible for monitoring the
patients, all 688 people were sought and only 302 (43.9%) were located. Of these 302 patients
found, 272 received the results from the CCZ, but only 225 were treated. All patients were
located and received collectors were willing to perform the stool examination, but only 150
(49.7%) returned the collectors with the samples. It was observed that 113 reported the use
of the drug and 33 (29.2%) of them remained positive for schistosomiasis, suggesting possible
recontamination or treatment failure. Of the 29 who claimed not to have been treated, 16
confirmed the presence of the parasite and 13 negative results. The remaining eight people
didn’t remember if they had taken specific treatment. Conclusions: The municipality of Aracaju
follows the directives of the schistosomiasis control program. The Santa Maria populations
presented in the last survey coverage near the goal of 80%. The Schistosomiasis Control
Programs activities are included among the tasks of the Family Health Strategy (FHS) teams
and notification is kept updated. Nevertheless while the insertion in the FHSs has occurred,
there was miscommunication between the CCZ and the basic care that results in loss of
segment of patients. This was the biggest problem occurred for the assessment of PCE.
The main problems were related to location records of patients, since from the listing of
the CCZ, because it was not possible determine a portion of cases in the Basic Health Unit
to which the patient belonged. Another observation that impairs the effectiveness of the
program was the intense migration of the district, about 30% of patients had moved from the
neighborhood and were not followed, leaving doubt as to the outcome of cases. Although
many patients have received the test results and referred to having been treated, about
30% of them remain infected. This result points to problems of program effectiveness, since
they relate to patients and that most employees tend to worry about their health situation.
411
412
índICE dE aUtORES
(AuTHOR InDEX)
Aagaard-Hansen, J. .................................................................................................................................. 75
Abdala, Renato ........................................................................................................................................ 95
Abramo, Clarice ..................................................................................................................................... 161
Abreu, Mery Natali Silva ................................................................................................ 240, 259, 271, 311
Afonso, Ana Júlia Pinto Fonseca Sieuve ................................................................................. 349, 392, 393
Agboola, Kolade M. ............................................................................................................................... 387
Aguiar, Alcino Palermo de ..................................................................................................................... 366
Aires, André de Lima ...................................................... 120, 178, 293, 299, 301, 302, 324, 328, 339, 354
Albuquerque, Eduardo .......................................................................................................................... 206
Albuquerque, Eduardo N. ...................................................................................................................... 282
Albuquerque, Jones ....................................................................................................................... 159, 254
Albuquerque, Mônica Camelo Pessoa de ............... 120, 178, 293, 299, 301, 302, 324, 328, 339, 354, 397
Alcântara, Bruna Cunha de ............................................................................................................ 154, 368
Alcântara, Leda Maria............................................................................................................................ 344
Alecrim, Vinícius Martins....................................................................................................... 179, 316, 340
Alencar, Alba Cristina Miranda de Barros ...................................................................................... 334, 336
Alencar, Carlos Henrique Morais de ...................................................................................................... 257
Almeida Júnior, Antônio Sérgio Alves de ............................................................................................... 315
Almeida, Giulliana T. ................................................................................................................................ 82
Almeida, José Antônio Pacheco de ................................................................................ 163, 215, 241, 269
Almeida, José Roberto de ...................................................................................................................... 322
Almeida, Luisa Maria Silveira de............................................................................................................ 174
Almeida, Luiza F. A. .......................................................................................................................... 80, 369
Almeida, Maria Cecília ................................................................................................... 238, 275, 344, 345
Almeida, Marta Martins ................................................................................................................ 212, 226
Almeida, Mônica Maria de .................................................................................................................... 345
Almeida, Roberta Cavalcanti de............................................................................................................. 322
Almeida, Roque Pacheco de .................................................................................................................. 241
Almeida, Thais Cavalcanti de ................................................................................................................. 322
Almeida, Thaís Helena Guilherme de .................................................................................................... 192
Almeida, Thays Miranda ................................................................................................................ 328, 339
Almeida, Vitor Alexandre Kessler de ..................................................................................................... 165
Alvarez, Maria Carolina Barbosa ........................................................................................................... 362
Alves, Clarice Carvalho .......................................................................................................................... 347
Alves, Luciana Patricia Lima............................................................................146, 204, 212, 220, 226, 305
Alves, Marcelo Ribeiro........................................................................................................................... 409
Alves, Maria Sonia Correia .................................................................................................................... 181
Alves, Marina Amaral ............................................................................................................................ 133
Alves, Renata de Sousa .......................................................................................................................... 260
Amante, Carlos Antônio ........................................................................................................................ 286
Amaral, Luciana Silva do ........................................................................................................................ 346
Amarante, Anderson Mendonça ........................................................................................................... 366
Amorim, Fábio Jorge Ramalho............................................................................................................... 269
Amorim, Fábio Ramalho de ................................................................................................................... 163
Anderson, Timothy J. C. ........................................................................................................................... 98
Andrade, Milton Hércules Guerra de .................................................................................... 128, 386, 388
413
Andrade, Mônica Coelho ....................................................................................................................... 192
Andrade, Rafael Cesar Lima Pedroso de ................................................................................................ 284
Andrade, Zilton de Araújo ..................................................................................................... 139, 177, 387
Aníbal, Fernanda de Freitas ............................................................................292, 304, 319, 335, 353, 399
Anjos, Zilma Perreira dos ....................................................................................................................... 324
Antunes, Carlos Aleberto....................................................................................................................... 366
Antunes, Carlos Mauricio de Figueiredo ........................................154, 234, 308, 312, 321, 330, 363, 387
Araújo, Emília Souza .............................................................................................................. 184, 211, 363
Araújo, Heloísa Sobreiro Selistre de ...................................................................................... 292, 335, 353
Araújo, Juliano Michel de .............................................................................................................. 118, 148
Araújo, Karina Conceição Gomes Machado de......................................243, 251, 252, 258, 280, 281, 411
Araújo, Lucas Henrique Lobato de........................................................................................................... 74
Araújo, Maria Ilma ....................................................................................................94, 122, 126, 337, 344
Araújo, Mykaella Andrade de ................................................................................................................ 278
Araújo, Roni Evêncio de ......................................................................................................... 315, 318, 359
Araújo, Rosiane...................................................................................................................................... 248
Araújo, Sidcley Bernardino de ............................................................................................................... 120
Araújo, Wildo Navegantes de ................................................................................................................ 249
Arruda, Nara Pedrosa ............................................................................................................................ 282
Assis, Thiago de Oliveira ........................................................................................................................ 275
Augusto, Ronaldo de Carvalho .............................................................................................. 228, 232, 236
Bafica, Aline Michelle Barbosa .............................................................................................................. 126
Bandeira, Angela Pontes................................................................................................................ 156, 310
Barbosa, Constança Simões...................................... 53, 109, 154, 243, 251, 252, 254, 267, 268, 281, 284
Barbosa, Luciene ................................................................................................................................... 411
Barbosa, Lúcio M. .................................................................................................................................... 86
Barbosa, Verônica Santos ...................................................................................................................... 251
Barboza, Delmany Moitinho .................................................................................................................. 215
Barboza, Morgana M. O......................................................................................................................... 262
Barnes, Kathleen C. ................................................................................................................................. 94
Barreiro, Eliezer J. .................................................................................................................................. 133
Barreto, Ana Virgínia Matos Sá...................................................................................... 152, 179, 316, 340
Barreto, Magali Gonçalves Muniz.................................................................................. 197, 273, 276, 288
Barros, André......................................................................................................................................... 335
Barros, Andréia Ferreira de ................................................................................................... 315, 318, 359
Barros, Giulliano Vilela .......................................................................................................................... 198
Barroso, Nathalia Vasconcelos .............................................................................................................. 411
Bassoli, Wenderson ............................................................................................................................... 234
Bastos Filho, Carmelo J. A. ..................................................................................................................... 159
Bastos, Dńarte Hermogenes................................................................................................................. 238
Bastos, Jairo K. ............................................................................................................................... 187, 298
Batista, Flávia Serrano ........................................................................................................................... 286
Beck, Lilian Christina Nóbrega Holsbach ................................................................................................. 76
Beckedorff, Felipe C. F.............................................................................................................................. 82
Belo, Silvana .......................................................................................................................................... 392
Belo, Silvana Maria Duarte .................................................................................................................... 105
Benitez, Osvaldo D. ................................................................................................................................ 142
Benjamim, Cláudia Farias ...................................................................................................................... 366
Bento Neto, Aloísio Ferreira .................................................................................................................. 411
Bergquist, Robert..................................................................................................................................... 70
414
Berriman, Matthew ................................................................................................................................. 98
Bertão, Humberto Gonçalves ................................................................................................................ 397
Bethony, Jeffrey Michael ........................................................................................................... 71, 74, 125
Bezerra, Fernando Schemelzer de Moraes ............................. 113, 186, 192, 193, 194, 257, 260, 262, 358
Bezerra, Lidya Ângelo ............................................................................................................................ 243
Blank, Walter A. ....................................................................................................................................... 86
Blanton, Ronald E. ................................................................................................................................... 86
Bocanegra, Silvana................................................................................................................................. 159
Boissier, Jérome ..................................................................................................................................... 130
Borda, C. Edgardo .................................................................................................................................. 142
Borges, Denise de Assuncão .................................................................................................................. 276
Borges, William de Castro ......................................................................................128, 368, 372, 386, 388
Botelho, Maria Ceci Salcedo .................................................................................................................. 291
Botelho, Patrícia Passos ......................................................................................................................... 234
Bozza, Marcelo Torres ................................................................................................................... 132, 360
Braga, Fernão Castro ............................................................................................................................. 184
Braga, Lidiane Bento .............................................................................................................................. 217
Brigatto, Olinda Mara ............................................................................................................................ 368
Brindley, Paul J. ........................................................................................................................................ 84
Buchmann, Fábio Fiebrig ....................................................................................................................... 265
Cabral, Fernanda J. ................................................................................................................................ 372
Caby, Stéphanie ............................................................................................................................... 80, 369
Caggegi, Andréia Aparecida................................................................................................................... 107
Calado, Maria Manuela P. ...................................................................................................................... 393
Caldeira, Roberta Lima .................................................................................................... 89, 137, 172, 217
Calefi, Paulo Sergio ................................................................................................................................ 187
Campos, Cristiane de Carvalho .............................................................................................. 188, 376, 378
Campos, Jonatan Marques ............................................................................................................ 386, 388
Campos, Julyana Viégas......................................................................................................................... 284
Cantanhede, Selma Patricia Diniz ...........................................................146, 204, 212, 220, 226, 290, 305
Capettini, Luciano S. A. .......................................................................................................................... 211
Cardoso, Andréa Valença............................................................................................................... 280, 411
Cardoso, Fernanda ................................................................................................................................ 125
Cardoso, Fernanda Sandes .................................................................................................................... 169
Cardoso, Luciana Santos .........................................................................................122, 126, 141, 337, 344
Carlôto, Aline Eduardo .................................................................................................................. 245, 296
Carmo, Theomira M. ............................................................................................................................... 86
Carneiro, Nidia Francisca de Figueiredo ................................................................................................ 111
Carneiro, Teiliane Rodrigues ...........................................................113, 186, 192, 193, 194, 260, 262, 358
Carneiro, Vitor Coutinho ....................................................................................................... 131, 132, 366
Carvalho, Andréa Teixeira ........................................................................................................ 95, 345, 352
Carvalho, Antonio P. .............................................................................................................................. 266
Carvalho, Camila Dantas de ................................................................................................................... 222
Carvalho, Edgar Marcelino de ..................................................................................94, 122, 126, 337, 344
Carvalho, Gabriel Costa de .................................................................................................................... 185
Carvalho, Gardênia Braz Figueiredo de ........................................................................................ 151, 362
Carvalho, Omar dos Santos ......................................................................................89, 137, 140, 217, 246
Carvalho, Tânia Antunes ........................................................................................................ 157, 165, 312
Carvalho, Tania Gomes de ..................................................................................................................... 235
Carvalho, Técia Maria Ulisses de ........................................................................................................... 196
415
Carvalho, Tiago Pinheiro Vaz de ............................................................................................................ 281
Carvalho, Vera Cristina Oliveira de ........................................................................................................ 276
Cassago, Alexandre ................................................................................................................................ 371
Castelo-Branco, Moragana T. L. ............................................................................................................. 343
Castro, Aline Pereira .............................................................................................................................. 198
Castro, Ana Karine Sarvel de ................................................................................................................. 239
Castro, Célia Maria Machado Barbosa de ..................................................................................... 328, 339
Castro, Luciana Lisboa Mota e ............................................................................................................... 311
Castro, Milton F. ............................................................................................................................ 140, 161
Cavalcanti, Marta Guimarães ................................................................................................................ 197
Cavalncanti, Natália Lima ...................................................................................................................... 397
Chagas, Anny Caroline Porto ................................................................................................................. 163
Chalmers, Iain W. ................................................................................................................................... 383
Chaves, Bruna Assis ............................................................................................................................... 154
Chaves, Carmem de Castro.................................................................................................................... 365
Chaves, Maria Elizabeth Cavalcanti ....................................................................................................... 397
Choi, Steve S. ......................................................................................................................................... 387
Christe, Rafael de Oliveira ..................................................................................................................... 107
Ciuffi, Kátia Jorge ................................................................................................................................... 187
Clemente, Isabel .................................................................................................................................... 392
Coelho, Kellen Rosa ............................................................................................................................... 271
Coelho, Pablo Menezes ......................................................................................................................... 218
Coelho, Paulo Marcos Zech .............................................. 66, 111, 118, 135, 148, 172, 174, 176, 188, 190
199, 211, 297, 360, 362, 376, 378, 394
Coimbra, Elaine Soares .......................................................................................................... 140, 161, 185
Coimbra, Mariana.................................................................................................................................. 197
Coimbra, Roney S. ........................................................................................................................... 85, 405
Coimbra, Thaissa Melo Galante..................................................................................................... 123, 150
Compagnon, Milton Cezar ..................................................................................................................... 365
Conceição, Jenisson Oliveira.................................................................................................................. 242
Conceição, Maria José ................................................................................................................... 245, 296
Corrêa Junior, Ary .................................................................................................................................. 211
Corrêa, Christiane Leal .......................................................................................................................... 332
Corrêa, Lygia dos Reis ............................................................................................................................ 144
Correia, Ricardo de Oliveira ........................................................................................... 292, 304, 319, 353
Cosseau, Céline ....................................................................................................................... 80, 130, 369
Costa Junior, Admar Borges da .............................................................................................................. 322
Costa-Silva, Matheus Fernandes ........................................................................................................... 352
Costa, Fernanda S. ................................................................................................................................. 363
Costa, Ivete Conchon ............................................................................................................................. 295
Costa, Jackson M. .................................................................................................................................... 86
Costa, Letícia Campos da ............................................................................................................... 327, 343
Costa, Marta Julia Faro dos Santos ....................

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