Desempenho de sementes peliculizadas de feijão

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PERFORMANCE OF FILM COATED COWPEA SEEDS DURING
STORAGE
Desempenho de sementes peliculizadas de feijão-caupi durante o armazenamento
RHAFAELA MACEIÓ DA SILVA
FRANCISCO DE ASSIS CARDOSO ALMEIDA
BRUNO ADELINO DE MELO
JOSIVANDA PALMEIRA GOMES
ACÁCIO FIGUEIREDO NETO
ESTHER MARIA BARROS DE ALBUQUERQUE
ABSTRACT
RESUMO
The aim of the present study is to evaluate the
performance of cowpea seeds film-coated with
polymer, manioc starch and polyvinyl acetate
(white glue) in the percentage of 25 to 75%,
including an aqueous extract (black pepper) and a
fungicide (carboxin+thiran), considering the
following: water soaking, germination and
accelerated aging during storage. The experiment
was divided into two phases. In the first phase,
the curves showing the water soaking capacity of
coated and treated seeds were studied. In the
second phase, the physiological quality of the
seeds was examined during storage at ambient
conditions. The experiments were arranged into
a randomized design. The storage experimental
data were subjected to analysis of variance by the
F test (p <0.05) and the averages, when required,
were compared by the Scott-Knott test (p <0.05).
As to the soaking curves, the data were subjected
to regression analysis of variance, producing,
consequently, the models for each treatment. The
water soaking curves for cowpea seeds displayed
a triphasic pattern. The application of coating on
to the cowpea seeds did not affect the
germination process. Higher germination during
storage process has been noticed in the
treatments: polymer + extract (P + E) and glue +
fungicide (C + F).
Objetivou-se, com este estudo, avaliar o
desempenho de sementes de feijão-caupi
peliculizadas com polímero, fécula de mandioca e
acetato de polivinila (cola branca), nos
percentuais de 25 e 75%, incorporando um
extrato aquoso (pimenta-do-reino) e um fungicida
(carboxin+thiran), quanto à embebição de água;
germinação e envelhecimento acelerado durante
o armazenamento. O trabalho foi realizado em
dois experimentos, estudando-se no primeiro as
curvas de embebição de água pelas sementes
peliculizadas e tratadas, e no segundo a
qualidade fisiológica durante o armazenamento,
em condições ambientais, destas mesmas
sementes. Os experimentos foram organizados
em delineamento inteiramente casualizado. Os
dados do experimento de armazenamento foram
submetidos a análise de variância pelo teste F (p
< 0,05) e as médias, quando necessário,
comparadas pelo teste de Scott-Knott (p < 0,05).
Para as curvas de embebição, os dados foram
submetidos à regressão na análise de variância,
gerando-se os modelos para cada tratamento. As
curvas de embebição de água para sementes de
feijão-caupi apresentaram padrão trifásico. A
aplicação das coberturas nas sementes de feijãocaupi não interferiu no processo de germinação.
A
maior
germinação,
ao
longo
do
armazenamento, foi constatada nos tratamentos
polímero + extrato (P + E) e cola + fungicida (C +
F).
KEYWORDS
COATING, CONSERVATION,
IMBIBITION, PHYSIOLOGICAL QUALITY.
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PALAVRAS-CHAVE
CONSERVAÇÃO, EMBEBIÇÃO,
QUALIDADE FISIOLÓGICA,
RECOBRIMENTO.
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INTRODUCTION
The main concern of producers in Brazil is to secure seed integrity. Seed treatment is
common agronomic accepted practice for most species. On the other hand, this practice is
inadequate because of product diversity and the health risks it poses to workers: the dust and
handling of highly toxic products (Queiroga et al. 2012b).
Seed coating is seen as a practical solution. This process consists of applying a liquid
film, usually, on single layers without altering seed weight and shape, ensuring, at the same time,
optimum adhesion and distribution of the active ingredients derived from the treatment. Besides,
it also facilitates identification and visual traceability (Gadotti & Puchala, 2010).
According to Avelar et al. (2011), the use of high quality seeds is widely recommended
by producers as one of the most effective means of minimizing costs and risks. The same authors
say that the polymer industry has progressed very fast in recent years, producing polymers
compatible with the formulations of conventional seed treatment. They also state that polymers
help preserve good seed appearance, making seeds look more colorful and glossy. Producers will
then provide seeds with higher physical and physiological integrity, making it possible to develop
more uniform seedlings.
Film coating, as a common technique, presents certain peculiarities that require careful
investigation in order to be adopted more frequently by producers (Pereira, 2005). Despite the
increased use of film-coated seeds, observed in recent years, there is little information available in
the literature on the behavior of these seeds during storage (Oliveira et al. 2003).
Storage is one of the major constraints on seed preservation. It is influenced by various factors,
such as initial seed quality, action of microorganisms and insects, types of packaging and duration
of storage (Almeida et al 2005; 2009; Queiroga et al. 2012a).
Given the above, and the lack of information on major crops, especially that of the
cowpea, it is our aim to evaluate the performance of film coated cowpea seeds with polymer,
starch and polyvinyl acetate (white glue) at concentrations of 25 and 75%, in combination with
the use of a plant extract (black pepper) and a fungicidal (thiran carboxin +) as a result of water
soaking: germination and accelerated aging during storage.
RESEARCH DEVELOPMENT
The experiment was conducted in the Storage and Processing Laboratory of Agricultural
Products (Lappa) at the Academic Agricultural Engineering Unit (UAEA) of the Center for
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Technology and Natural Resources (CTRN) of the Federal University of Campina Grande
(UFCG).
The seeds of cowpea [Vigna unguiculata (L) Walp] were obtained from local producers
in Campina Grande, PB. The seeds came from the 2012/2013 harvest. The initial quality of the
seeds was evaluated by determining their water content, germination and vigor (Brazil, 2009).
The product used for film coating the seeds was the polymer POLIFIX DJ-G4, obtained from
the company LABORSAN® - Trade and Dyes Imports and Polymers Ltd. (liquid formulation
product).
The manioc starch and the polyvinyl acetate (glue) were purchased from special stores in
Campina Grande, PB. The starch was prepared as recommended by the Fakhouri methodology
(2007). After reaching its gelatinous point, the starch was cooled and diluted in percentages of
25 and 75%, with an addition of 20 ml of food coloring. The aqueous extract of black pepper
(Piper nigrum L.) was obtained by using the methodology of Almeida et al. (2009), and the
fungicide employed was carboxin + thiran, acquired from a local store. For both treatment
products (natural and synthetic), an amount corresponding to 25% of the syrup volume was
used. The seed film coating was obtained with the use of different syrups made-up of the proper
materials, as shown in Table 1.
The work included two experiments. In the first experiment, we studied the water
imbibition curves as presented by the film-coated seeds at concentrations of 25 and 75%. For
this, the initial mass of the film-coated seeds was determined. The seeds were placed on a wire
mesh and stored in plastic containers with 200 ml of distilled water. Then the containers were
closed with a lid and taken to a germination chamber type B.O.D at constant temperature of 10°
C. The seed masses were measured at intervals of 24-h until they reached the necessary moisture
to trigger germination process. For each material, 200 g of seeds were film coated.
In the second experiment, the storage of the treated film-coated seeds was carried out
under ambient conditions of temperature and relative humidity, in pet-type packaging
(polyethylene) with capacity of 250 g, for a period of 120 days. During storage time, the seeds’
physiological quality was verified every 30 days by the tests described below:
Initial water content: determined by the standard oven method according to the equation found
in the Rules for Seed Analysis (Brazil, 2009).
Germination test: conducted with four subsamples of 50 seeds, using as subtract
vermiculite moistened with distilled water till it reaches 60% of its holding capacity.
Germination evaluation was made on the eighth day following sowing, considering the number of
normal seedlings that have emerged (Brazil, 2009).
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Accelerated aging test: the methodology described by Mark Son (1994) was used, with
modifications, and consisted of selecting a seed sample for each treatment. The samples were
distributed along a single layer, placed on a plastic mesh, stored in plastic boxes containing 200
ml of distilled water, observing a distance of approximately 2 cm between the water level and the
seeds. Then, the boxes were sealed and taken to a B.O.D. germination chamber, set to work at
temperature of 42 ° C for 72 h. After this period, 200 seeds were chosen randomly and divided
into four subsamples of 50 seeds each. These were then submitted to a germination test.
The experimental project was entirely randomized. The cowpea storage experiment was
arranged along a factorial 5 x 2 x 4 (film coating materials x concentrations of materials x storage
periods). The data were subjected to analysis of variance (P <0.05) and the means, when
exhibiting significant difference, were matched by the Scott-Knott test (P <0.05) using the
Assistat software version 7.6 (Silva & Azevedo, 2009). As to the study of water uptake by the
seeds, the data were then subjected to regression in the analysis of variance and duly checked by
using the statistical program Bioestat 5.0.
Figure 1A shows that the curve of water absorbed by the cowpea seeds in the material
interaction of film coating and the period (days) follows a three-phase standard element
proposed by Bewley & Black (1994), where the first interval (Phase I) lasted 10 days as from the
beginning of the experiment. The film coated seeds absorbed 23.06% humidity for 10 to
33.06% of moisture. From the first interval up to the second interval (phase II), which lasted 12
days, the seeds absorbed only 11.94% moisture, almost half of the retained moisture in Phase I,
which was completed two days earlier than the expected time. In Phase III, this occurred 22 days
after the beginning of the experiment. The absorption of water by the seeds is smaller compared
to that of the first two phases; and it becomes smaller and smaller as soaking time moves forward.
At this stage, which lasted eight days, the seeds had their moisture content increased by only 3%.
Studies by Peske & Peske (2011) observed the total amount of water absorbed by the seeds
during imbibition. This generally does not exceed two to three times the mass of dry seed. Then,
to initiate the process of corn seed germination, for instance, the soaking process is expected to
hold 30 to 35% of water in relation to dry mass. In the case of soya beans, this must be around
50%.
In all treatments, the imbibition process studied was characterized by a rapid absorption
of water followed by a sharp reduction in the imbibition rate. On obtaining water content similar
to that of the seed physiological maturity, the seed again presents pronounced water absorption
speed but at much lower rate. These last two stages are fully described by Bewley & Black (1994).
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They say that, in phase II, the intensive transportation of substances broken in phase I shifts from
the reserve tissue on to the meristematic tissue. In phase III, the substances that have been
transported during the second stage rearrange to form the cell wall.
Almeida & Rock (2008) noticed that the use of waxes for multilayer seed coating makes
seeds resistant to water absorption, delaying, in this way, the development of seedlings. However,
highly hygroscopic polymers do not affect these features, which are in agreement with the data
obtained for the present work, for which the materials used, including a polymer, do not prevent
and/or inhibit water absorption by film-coated seeds.
Regression analyses of the film coating material shown in Table 2, demonstrated that the
coefficients of determination ranged from 93 to 99%, indicating that the models had been
satisfactorily adjusted to the experimental data, and that they can be used to estimate the intervals
between the periods studied.
It is clear that the soaking process has initially exhibited a water content of 11.04%.
Besides, the film coating does not prevent the soaking of the cowpea seeds. These results are in
tune with those found by Huth et al. (2013) who observed that the film coating process did not
affect the capacity of water absorption by soybean seeds.
The analysis of variance has produced some highly significant effects for all factors (film-coating
material, concentration and time), and also for the dual interaction between film-coating material
and storage period (Table 3).
Comparing the film-coating material at each storage period, it appears that, in 30 days,
the germination of film-coated seeds with polymer + fungicide, polymer + plant extract and cola
+ fungicide showed no statistical difference, being such treatments the ones that presented the
highest germination percentage (97.25, 95.00 and 94.75%, respectively). On the other hand,
seeds with no treatment and those starch-encapsulated + those treated with fungicide had lower
germination; ranging from 82.5 to 88.75% (Table 4).
For sixty-day storage, the film coating polymer + plant extract and cola + fungicide
produce seeds with germination of 91.25 and 95.00%, causing germinations to be much higher
and statistically equal to each other. The lowest germination was obtained when the seeds were
film-coated with starch + fungicide (73.00%). The seeds of the witness and those film-coated
with polymer + fungicide presented intermediate performance, with germination ranging from
78.50 to 83.25%.
At 90 and 120-day storage, one can see that the highest germination percentage occurred
in film-coating seeds with plant extract and polymer + glue + fungicide, with germination of
78.75 and 82.00%, respectively. Witness, during these two periods, showed no germination
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(0.00%). The film-coating polymer + fungicide and starch + fungicide granted the seeds an
intermediate performance, with germination of 63.25 and 68.00%, respectively.
Comparing storage periods within each film-coating material, it becomes apparent in the
witness that within periods of 30 and 60 days, germination was found to range from 82.50 to
78.50%, being not much statically different. However, the germination within 30 and 60 days of
storage was statistically different from that observed at 90 and 120 days of storage (0.00%).
With the seeds coated with polymer + fungicide, all four storage periods revealed no
statistical difference between them. They presented better germination after 30 days of storage
(97.25%), and the lowest germination was found to occur after 120 days of storage (63.25%).
The cowpea seeds film-coated with polymer + vegetal extract, at 30, 60 and 90 days of storage
showed no statistical difference with germination ranging from 87.75 to 95.00%. Germination
along these periods was statistically different from germination observed at 120 days of storage
(78.75%), being the lowest of all periods.
When the seeds were film-coated with starch + fungicide, the highest germination was
noticed to occur at 30 days (88.75%), differing considerably from germinations that occurred at
60, 90 and 120 days (73.00; 73.25 and 68 00%, respectively), which were statistically equal to
each other.
It has been observed that by using glue + fungicide higher germination occurred at 30
and 60 days (94.75 and 95.00%, respectively) – statistically equal to each other and different
from germinations observed at 90 and 120 days of storage (82.00 and 82.00%).
Pereira et al. (2011), researching into soybeans in storage, found that film-coating
associated with fungicide did not affect the physiological quality of seeds. They also observed
that germination of film-coating seeds was higher than in non-film coating seeds. These results
support the present study.
This assertion is consistent with a work by Pereira et al. (2009), in which the authors
concluded, after investigating the association of fungicides and polymers, that seed physiological
quality was not affected by the association of these products. This corroborates with the results
of the present study, and is in tune with the statements by Carvalho et al. (2010) that filmcoating is a satisfactory method for the distribution and delivery of chemicals into the seeds; their
use being not harmful to germination, allowing the formation of a protective film against adverse
environmental conditions.
Table 5 shows the means of post-accelerated-aging germination of cowpea seeds filmcoated with polymer, manioc starch and polyvinyl acetate, plus the incorporation of a vegetable
extract of black pepper and fungicide during storage. By comparing the film-coating products at
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each storage period, it became apparent that at 30 days, vigor was at its highest when the seeds
were coated with polymer + fungicide (87.25%), polymer + extract plant (91.00%), cola +
fungicide (88.00%) and starch + fungicide (88.25 %). These results are statistically equal to
each other, but different from the witness, the whose vigor was 80.00%, being the lowest among
the averages for this period.
After 60 days of storage, the materials that presented better performance were: polymer
+ plant extract (81.50%), cola + fungicide (83.00%), which were statistically equal to the
control (78.00%). On the other hand, when the seeds were coated with polymer + fungicide and
starch + fungicidal, vigor was 71.50 and 72.50%, respectively, being mutually the same and
different from the other treatments.
At 90 days of storage, the seeds film-coated with glue and starch and fungicide +
fungicide showed better performance post-accelerated aging with vigor of 81.75 and 86.25%,
respectively. When using the polymer + polymer + fungicide + vegetal extracts, the seeds had an
average reduction of 20% compared to the two previous treatments, differing statistically from
those. The witness had the lowest place among the treatments (7.00%).
After a 120-day storage, it was observed that film-coating of seeds with glue + fungicide
presented the greatest effect among the materials (82.25%), differing significantly from results
when polymer + fungicide (57.50%), polymer + vegetal extract (61.75%) + fungicide and
starch (65.00%) were used. These were statistically equal to each other. The seeds of the witness,
at 120 days, however, showed no vigor.
Based on the results obtained, one may say that film-coating does not affect germination
and seed vigor. Moreover, the association of fungicide carboxin + thiran and black pepper
extract, are now safely applicable to storage. Similar results were found by Carvalho et al. (2010)
who concluded that film-coating is a suitable method for the distribution and use of chemicals to
seed preservation.
CONCLUSIONS
1. The water soaking curves for the cowpea seeds presented a triphasic pattern;
2. Application of coatings on cowpea seeds did not affect germination and vigor;
3. The glue and fungicide treatment (C + F) preserves the seed quality during storage.
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(2013). Velocidade de embebição de sementes de soja submetidas a diferentes recobrimentos. Informativo
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Table 1. Composition of syrups used in film coating/treatment * of cowpea seeds.
Treatments
Material (%)
Treatments* (%)
Water (%)
Witnesses
0
0
0
Polymer 25% + fungicide
25
25
50
Polymer 75% + fungicide
75
25
0
Polymer 25%+ extract
25
25
50
Polymer 75%+ extract
75
25
0
Starch 25% + fungicide
25
25
50
Starch 75% + fungicide
75
25
0
Glue 25% + fungicide
25
25
50
Glue 75% + fungicide
75
25
0
* Black pepper extract and the fungicide carboxin + thiram were used at a ratio of 25% of spray
volume. The total volume of syrup used corresponded to 1% of the total weight of the seeds.
Table 2. Regression equation and coefficient of determination of soaking water capacity for
cowpea seeds after film-coating and treatment with black pepper extract plus fungicide
carboxin + thiran.
Coating material
Regression equations
Witness
y = 11,6779x2** + 2,7343x** - 0,0531**
Polymer 25% + fungicide
y = 12,7622x2** + 2,3712x** - 0,0439**
Polymer 75% + fungicide
y = 13,026x2** + 2,2301x** - 0,0391ns
Polymer 25% + extract
y = 11,231x2** + 2,5532x** - 0,0463**
Polymer 75% + extract
y=11,897x2** + 2,5854x** - 0,0466**
Starch 25% + fungicide
y = 15,765x2** + 2,0200x** - 0,0392**
Starch 75% + fungicide
y = 16,275x2** + 2,0849x** - 0,0415**
Glue 25% + fungicide
y = 17,004x2** + 1,871x** - 0,0322**
Glue 75% + fungicide
y = 17,320x2** + 2,0604x** - 0,0366**
*Substantial at 1% probability (p <0.01); Non-substantial (p ≥ 0.05); * Substantial 0.05
probability. (0.01 p ≤ 0.05).
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(R²)
0,98
0,98
0,97
0,98
0,99
0,95
0,94
0,93
0,95
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Table 3. Summary of variance analyses, suggesting the degree of freedom (DF) and the mean
square (MS) for germination and accelerated aging of film-coated cowpea with polymer,
starch and glue at different concentrations, treated with black pepper extract1 and
fungicide carboxin + thiran2.
Germination
Accelerated aging
FV
GL
QM
GL
QM
Coated material (CM)
4
12702,83**
4
8857,00**
Concentration (C)
1
756,9**
1
518,40**
Time (T)
3
10116,2**
3
9197,20**
MP x C
4
103,71ns
4
62,65ns
MP x T
12
2565,05**
12
2459,86**
CxT
3
38,56ns
3
48,85ns
MP x C x T
12
20,96*
12
2405,69**
Treatments
39
2909,73**
39
58,53
Residue
120
61,73
120
Total
159
159
** Significant 0.01 probability (p <0.01); nsnon-significant (p ≥ 0.05); 0.05 * Significant 0.05
probability (p ≤ 0.01 0.05); 1,2black pepper extract and fungicide carboxin + thiran were used
at a ratio of 25% of the applied syrup volume.
Table 4. Germination average values (%) of cowpea seeds for interaction *treatments in time
Storage time (days)
Treatments
30
60
90
120
Witness
82,50 bA
78,50 bA
0,00 cB
0,00 cB
Polymer + fungicide
97,25 aA
83,25 bB
76,75 bC
63,25 bD
Polymer + extract
95,00 aA
91,25 aA
87,75 aA
78,75 aB
Starch + fungicide
88,75 bA
73,00 cB
73,25 bB
68,00 bB
Glue + fungicide
94,75 aA
95,00 aA
82,00 aB
82,00 aB
The means followed by the same capital letters in the columns are statistically different from each
other at 5% probability according to the Scott-Knott test. * Black pepper extract and fungicide
carboxin + thiran was used in the ratio of 25% of the applied syrup volume; initial germination
at 90%, and initial moisture content 12.5%; CV = 10.54%.
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Table 5. Average values of accelerated aging (%) of cowpea seeds for the interaction *treatments
at periods
Storage time (days)
Treatments
30
60
90
120
Witness
80,00 bA
78,00 aA
7,00 cB
0,00 cB
Polymer + fungicide
87,25 aA
71,50 bB
65,75 bB
57,50 bC
Polymer + extract
91,00 aA
81,50 aB
66,75 bC
61,75 bC
Starch + fungicide
88,00 aA
83,00 aA
81,75 aA
82,25 aA
Glue + fungicide
88,25 aA
72,50 bB
86,25 aA
65,00 bB
The averages followed by the same capital letter in the rows and lowercase in the columns are not
statistically different from each other at 5% probability according to the Scott-Knott test. * The
black pepper extract and the fungicide carboxin + thiran were used in the ratio of 25% of the
volume of the applied syrup. Initial aging was 76.00% and initial moisture was 12.5% CV =
10.97%.
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Figure 1. Imbibition curves of cowpea seeds after film-coating and treatment. A. Polymer 25%
+Fungicide; B. Polymer 75% + Fungicide; C. Polymer 25% + Extract; D. Polymer + 75%
extract; E. starch 25% + Fungicide; F. starch 75% + Fungicide; G. Glue 25% + Fungicide; H.
Glue 75% + Fungicide and I. Witness. Initial moisture 12.5%.
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Raphaela Maceió da Silva
Possui graduação em Agronomia pela Universidade Federal Rural
de Pernambuco. Atualmente é aluna do Doutorado em
Engenharia Agrícola na Universidade Federal de Campina Grande
com ênfase em Processamento e Armazenamento de Produtos
Agrícolas.
Francisco de Assis Cardoso Almeida
Doutor em Agronomia pela Universidad Politécnica de Córdoba e
Pós-doutor em Agronomia pela Universidad Politécnica de
Madrid, Professor Titular da Universidade Federal de Campina
Grande.
Bruno Adelino de Melo
Possui Graduação em Agronomia e Mestrado em Engenharia
Agrícola pela Universidade Federal de Campina Grande.
Atualmente é aluno de Doutorado em Engenharia Agrícola pela
UFCG na linha de pesquisa de Processamento e Armazenamento
de Produtos Agrícola.
Josivanda Palmeira Gomes
Possui graduação e Mestrado em Engenharia Agrícola pela
Universidade Federal da Paraíba e doutorado em Engenharia de
Alimentos pela Universidade Estadual de Campinas (1999).
Professora Titular da Universidade Federal de Campina Grande.
Acácio Figueirêdo Neto
Possui Graduação em Engenharia Agronômica pela Universidade
Federal da Paraíba, Mestrado em Engenharia Agrícola pela
Universidade Federal de Campina Grande e Doutorado em
Engenharia Agrícola pelo convênio entre UFV/UFCG (2012).
Professor Adjunto da Universidade Federal do Vale do São
Francisco - UNIVASF.
Esther Maria Barros de Albuquerque
Graduada em Ciências Biológicas pela UEPB, Mestre em
Engenharia Agricola e Doutora em Engenharia de Processos pela
UFCG. Professora. E-mail: [email protected]
RECEBIDO EM 05/01/2016
APROVADO EM 14/03/2016
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