Effect of human hydrosalpinx fluid in fertilization rate of mouse oocyte and embryo quality.

OBJECTIVE: To determine whether human hydrosalpinx fluid might have a deleterious effect on the fertilization rate and embryonic development of the exposed mouse oocytes. METHODS: Mouse cumulus-oocyte complexes (COCs) were randomly allocated for exposure to pure hydrosalpinx fluid (100% HSF group, n=400), EBSS containing 50% of hydrosalpinx fluid (50% HSF group, n=320) and pure EBSS (control group, n=300). RESULTS: The results showed that the fertilization rate in the 100% HSF group was significantly lower than the control group (64.0% versus 73.0%, p=0.031). The blastocyst formation rate was also lower in the 100% HSF group than 50% HSF and the control group (51.5% versus 56.9% versus 56.3%, respectively), but not statistically significant (p=0.275). There was no significant difference in the mean numbers of cells in the ICM, TE, and total cell number in blastocysts from the control group and two hydrosalpinx fluid exposure groups. CONCLUSIONS: Human hydrosalpinx fluid has a negative effect on the fertilization rate of the exposed mouse oocytes. However, this effect was found only in undiluted concentration and does not affect the subsequence of embryonic development and blastocyst cell number.

Morphological analysis of Prochilodus argenteus ovaries in successful and unsuccessful hypophysation-induced reproduction.

Pampus argenteus is a species of economic importance for professional fishing and aquaculture. To analyse the quality of oocytes of P. argenteus, 10 females were kept in tanks of 200 m2 with an average depth of 1 m in the Integrated Center of Fishing Resources and Aquaculture of Três Marias - CODEVASF. Specimens were submitted to hypophysation-induced reproduction with crude extract of common carp pituitary (EBHC). Females received two doses (0.8-1.0 and 5.0-6.6 mg of EBHC/kg body weight, respectively), with a 12-h interval between doses. For males, a single dose (2.7 mg/kg body weight) was applied at the same time as the females' second dose. Oocytes were extruded manually 8 h after the second hormonal dose at 26°C. It was observed that seven females responded positively to the procedure while the other three released bloody and lumpy oocytes. For histological analysis, ovarian fragments were fixed in Bouin's liquid for 8-12 h and submitted to routine histological techniques. The protocol was considered successful for 70% of females and 100% of males. The fertilization rate of the females from the unsuccessful group was very low, and histological analyses showed that most of their oocytes were in follicular atresia, suggesting a delay in hormonal administration or extrusion could have occurred. Despite the hypophysation protocol being considered adequate for the induced reproduction of P. argenteus, complementary studies are necessary to evaluate the possible causes of this degenerative process.

Sex-specific differences in reproductive life-history traits of the moss Weissia jamaicensis.

PREMISE: We investigated sex-specific differences in the life-history traits of a metapopulation of the dioicous moss Weissia jamaicensis. Field observations revealed high rates of fertilization, which is uncommon for most dioicous bryophytes. We raised four hypotheses associated with the way the reproductive traits are related to the fertilization rate in this metapopulation. METHODS: We sampled 10 patches of the metapopulation and quantified sexual expression, sex ratio, reproductive success, and reproductive allocation. The ramets were classified as male, non-sporophytic female, sporophytic female, or non-sex-expressing. Thirty ramets from each of the categories expressing sex were placed for regeneration to test the effect of reproductive allocation on this trait. RESULTS: We found greater expression of the female function in all patches, implying a female bias in the metapopulation. The number of male ramets was variable in each patch and did not affect reproductive success. At the prezygotic level, the allocation of resources to the male function was higher. However, the large allocation of resources to sporophyte development in sporophytic females, which exceeded allocations at prezygotic levels, was related to the higher mortality rate of these ramets, suggesting reproductive cost. CONCLUSIONS: The prezygotic ramets that allocated the greatest amount of resources to reproduction expressed sex less frequently, biasing the sex ratio toward the sex that allocated the least amount of resources to reproduction. Overall, the ramets that allocated the greatest amount of resources to reproduction had the lowest regeneration rate, suggesting reproductive cost.

Dual trigger with gonadotropin-releasing hormone agonist and human chorionic gonadotropin significantly improves oocyte yield in normal responders on GnRH-antagonist cycles.

OBJECTIVE: During in vitro fertilization (IVF) cycles, final oocyte maturation is usually triggered by human Chorionic Gonadotropin (hCG) for its known effect in mimicking Luteinizing Hormone (LH) surge; however, with the widespread use of the 'antagonist protocol', Gonadotropin Releasing Hormone agonist (GnRHa) is being more commonly employed as a trigger in order to minimize or eliminate the risk of ovarian hyper-stimulation syndrome (OHSS). Many studies proved its efficacy in inducing oocyte maturation and its safety in preventing OHSS in high-risk groups. Moreover, some studies showed that GnRHa trigger may improve oocyte yield. This study aimed to further explore any beneficial effect of adding GnRHa to hCG (dual trigger) on oocyte yield and fertilization rate in normal responder women. METHODS: We retrospectively reviewed and analyzed the data from 127 patients on antagonist protocol (67 dual trigger and 60 HCG trigger). RESULTS: The number of total oocytes, the number of MII oocytes and the number of fertilized oocytes were all significantly higher with the dual trigger protocol compared to hCG-only trigger. However, there is no significant difference in clinical pregnancy rate. CONCLUSIONS: Using the dual trigger improved the number and quality of oocytes, and the fertilization rate in normal responders.

Inseminating dose for the artificial fertilization of Brycon amazonicus (Teleostei: Characidae).

Knowledge of the sperm-oocyte ratio in fish fertilization serves as the basis for studies on artificial reproduction and gamete manipulation. The aim of this study was to determine the minimum insemination dose for Brycon amazonicus oocyte fertilization. Female and male gametes were used and tested with the following doses of spermatozoa oocyte-1 ml-1: 10,000, 20,000, 40,000, 60,000 and 80,000 (in triplicate). Fertilization rates were calculated and estimated from the regression equation by applying the segmented regression model 'Linear Response Plateau' to determine the appropriate proportion of gametes. Based on the equation Y = 14.3415 + 0.0007836X, the fertilization rate increased up to 63.34% as it reached a plateau with a proportion of 62,524 spermatozoa oocyte-1 ml-1, which is the minimum insemination dose recommended for artificial insemination of the species.

Protein profile as a quality indicator of cryopreserved semen from Tambaqui Colossoma macropomum (Cuvier, 1818) / Perfil protéico como indicador qualitativo do semen criopreservado de Tambaqui Colossoma macropomum (Cuvier, 1818)

The aim of this study was to evaluate the association between proteins in the seminal plasma of tambaqui Colossoma macropomum (Cuvier, 1818) with seminal quality indicators after thawing. The semen was cryopreserved with a dilution based on BTS with 8% DMSO. A 200 µL sample of semen from each animal was diluted in 800 µL BTS, centrifuged at 800 rpm, and the supernatant was cryopreserved to further analyze of the protein profile of seminal plasma through one-dimensional electrophoresis (SDS-PAGE). After 15 days of cryopreservation, a cryopreserved semen straw was thawed to analyze both qualitative and quantitative parameters. When considering all collections, the SDS-PAGE identified 15 protein bands in the seminal plasma of tambaqui. When the interaction (presence or absence) between proteins observed in the seminal plasma and the post thawed spermatic parameters was evaluated, we observed a great influence of the presence of proteins on spermatic quality. A greater (P 0.05) fertilization rate was observed with the presence of proteins 12, 34, 44, 85, and 90 kDa. Proteins in seminal plasma of tambaqui influenced the spermatic quality after thawing, and thus, they can be utilized as an indicator of sperm quality, especially the proteins with a molecular weight 50 kDa.(AU)

O objetivo desse estudo foi de avaliar a associação entre a presença de proteínas no plasma seminal do tambaqui Colossoma macropomum (Cuvier, 1818) com indicadores de qualidade seminal pós-descongelamento. O semen foi criopreservado com diluidor a base de BTS com 8% DMSO. Uma amostra de 200 µL de semen de cada animal foi diluída em 800 µL de BTS, e centrifugada em 800 rpm, e somente o sobrenadante foi criopreservado para posterior análise do perfil proteico do plasma seminal, através da eletroforese unidimensional (SDS-PAGE). Decorridos 15 dias da criopreservação, uma palheta com semen criopreservado foi descongelado para análise dos parâmetros quali-quantitativos. Considerando todas as coletas, o SDS-PAGE identificou 15 bandas proteicas no plasma seminal do tambaqui. Quando se avaliou a interação (presença ou ausência) das proteínas encontradas no plasma seminal, com os parâmetros espermáticos pós-descongelamento, observou-se grande influência da presença das proteínas na qualidade espermática. Observou-se maior taxa de fertilização (P 0,05) com a presença das proteínas 12, 34, 44, 85 e 90 kDa. As proteínas do plasma seminal de tambaqui influenciaram na qualidade espermática após descongelamento, podendo ser utilizadas como indicadores para a qualidade espermática após descongelamento, principalmente as proteínas com peso molecular 50 kDa.(AU)

Protein profile as a quality indicator of cryopreserved semen from Tambaqui Colossoma macropomum (Cuvier, 1818) / Perfil protéico como indicador qualitativo do semen criopreservado de Tambaqui Colossoma macropomum (Cuvier, 1818)

Abstract The aim of this study was to evaluate the association between proteins in the seminal plasma of tambaqui Colossoma macropomum (Cuvier, 1818) with seminal quality indicators after thawing. The semen was cryopreserved with a dilution based on BTS with 8% DMSO. A 200 µL sample of semen from each animal was diluted in 800 µL BTS, centrifuged at 800 rpm, and the supernatant was cryopreserved to further analyze of the protein profile of seminal plasma through one-dimensional electrophoresis (SDS-PAGE). After 15 days of cryopreservation, a cryopreserved semen straw was thawed to analyze both qualitative and quantitative parameters. When considering all collections, the SDS-PAGE identified 15 protein bands in the seminal plasma of tambaqui. When the interaction (presence or absence) between proteins observed in the seminal plasma and the post thawed spermatic parameters was evaluated, we observed a great influence of the presence of proteins on spermatic quality. A greater (P<0.05) fertilization rate was observed with the presence of proteins 12, 34, 44, 85, and 90 kDa. Proteins in seminal plasma of tambaqui influenced the spermatic quality after thawing, and thus, they can be utilized as an indicator of sperm quality, especially the proteins with a molecular weight ≤ 50 kDa.

Resumo O objetivo desse estudo foi de avaliar a associação entre a presença de proteínas no plasma seminal do tambaqui Colossoma macropomum (Cuvier, 1818) com indicadores de qualidade seminal pós-descongelamento. O semen foi criopreservado com diluidor a base de BTS com 8% DMSO. Uma amostra de 200 µL de semen de cada animal foi diluída em 800 µL de BTS, e centrifugada em 800 rpm, e somente o sobrenadante foi criopreservado para posterior análise do perfil proteico do plasma seminal, através da eletroforese unidimensional (SDS-PAGE). Decorridos 15 dias da criopreservação, uma palheta com semen criopreservado foi descongelado para análise dos parâmetros quali-quantitativos. Considerando todas as coletas, o SDS-PAGE identificou 15 bandas proteicas no plasma seminal do tambaqui. Quando se avaliou a interação (presença ou ausência) das proteínas encontradas no plasma seminal, com os parâmetros espermáticos pós-descongelamento, observou-se grande influência da presença das proteínas na qualidade espermática. Observou-se maior taxa de fertilização (P<0,05) com a presença das proteínas 12, 34, 44, 85 e 90 kDa. As proteínas do plasma seminal de tambaqui influenciaram na qualidade espermática após descongelamento, podendo ser utilizadas como indicadores para a qualidade espermática após descongelamento, principalmente as proteínas com peso molecular ≤50 kDa.

Protein profile as a quality indicator of cryopreserved semen from Tambaqui Colossoma macropomum (Cuvier, 1818)

Abstract The aim of this study was to evaluate the association between proteins in the seminal plasma of tambaqui Colossoma macropomum (Cuvier, 1818) with seminal quality indicators after thawing. The semen was cryopreserved with a dilution based on BTS with 8% DMSO. A 200 µL sample of semen from each animal was diluted in 800 µL BTS, centrifuged at 800 rpm, and the supernatant was cryopreserved to further analyze of the protein profile of seminal plasma through one-dimensional electrophoresis (SDS-PAGE). After 15 days of cryopreservation, a cryopreserved semen straw was thawed to analyze both qualitative and quantitative parameters. When considering all collections, the SDS-PAGE identified 15 protein bands in the seminal plasma of tambaqui. When the interaction (presence or absence) between proteins observed in the seminal plasma and the post thawed spermatic parameters was evaluated, we observed a great influence of the presence of proteins on spermatic quality. A greater (P 0.05) fertilization rate was observed with the presence of proteins 12, 34, 44, 85, and 90 kDa. Proteins in seminal plasma of tambaqui influenced the spermatic quality after thawing, and thus, they can be utilized as an indicator of sperm quality, especially the proteins with a molecular weight 50 kDa.

Resumo O objetivo desse estudo foi de avaliar a associação entre a presença de proteínas no plasma seminal do tambaqui Colossoma macropomum (Cuvier, 1818) com indicadores de qualidade seminal pós-descongelamento. O semen foi criopreservado com diluidor a base de BTS com 8% DMSO. Uma amostra de 200 µL de semen de cada animal foi diluída em 800 µL de BTS, e centrifugada em 800 rpm, e somente o sobrenadante foi criopreservado para posterior análise do perfil proteico do plasma seminal, através da eletroforese unidimensional (SDS-PAGE). Decorridos 15 dias da criopreservação, uma palheta com semen criopreservado foi descongelado para análise dos parâmetros quali-quantitativos. Considerando todas as coletas, o SDS-PAGE identificou 15 bandas proteicas no plasma seminal do tambaqui. Quando se avaliou a interação (presença ou ausência) das proteínas encontradas no plasma seminal, com os parâmetros espermáticos pós-descongelamento, observou-se grande influência da presença das proteínas na qualidade espermática. Observou-se maior taxa de fertilização (P 0,05) com a presença das proteínas 12, 34, 44, 85 e 90 kDa. As proteínas do plasma seminal de tambaqui influenciaram na qualidade espermática após descongelamento, podendo ser utilizadas como indicadores para a qualidade espermática após descongelamento, principalmente as proteínas com peso molecular 50 kDa.

Avaliação de diferentes crioprotetores e tempos de descongelamento de sêmen de tambaqui Colossoma macropomum / Evaluation of different cryoprotectants and thawing times of tambaqui Colossoma macropomum semen

O objetivo com o trabalho foi avaliar a eficiência de quatro crioprotetores adicionados ao meio diluidor na criopreservação do sêmen de tambaqui (Colossoma macropomum) e melhor protocolo de descongelamento. Foram realizadas quatro coletas com intervalos regulares de amostras de sêmen (pool) de dez reprodutores. Os crioprotetores testados foram: glicerol, dimeltilsulfóxido (DMSO), dimetilformamida (DMF) e metanol, na proporção de 10,0%. Para a avaliação da qualidade do sêmen após o descongelamento foi realizado o teste in vivo das amostras em três protocolos diferentes para cada crioprotetor, com três tempos de descongelamento (15, 30 e 45s) na temperatura de 30ºC, totalizando 12 tratamentos com cinco repetições em esquema fatorial. Os resultados da fertilização foram submetidos a análise de variância (ANOVA) e teste de Tukey, com significância de 0,05. O tratamento que resultou em melhor taxa de fertilização foi o que continha metanol como crioprotetor e o descongelamento no tempo de 30 s, com taxa de fertilização média de 77,65%, sendo, entre os tratamentos avaliados, o mais recomendado para a criopreservação do sêmen de tambaqui.(AU)

The objective of the study was to evaluate the efficiency of four cryoprotectants added to the extender on sperm cryopreservation of tambaqui Colossoma macropomum and to define the best thawing protocol. Four semen samples were collected at regular intervals from ten breeding males (pool). The cryoprotectants tested were: glycerol, dimethylsulfoxide (DMSO), dimethylformamide (DMF) and methanol at a ratio of 10.0%. For the evaluation of semen quality after thawing in vivo test samples in three different protocols for each cryoprotectant was performed with three thawing times (15, 30 and 45s) at 30ºC temperature, totalizing 12 treatments with five replicates in factorial design. The results of fertilization were subjected to analysis of variance (ANOVA) and Tukey's test, with significance of 0.05. The treatment that resulted in improved fertilization rate was the one containing methanol as cryoprotectant and thawing time of 30 s, with a mean fertilization rate of 77.65%, being among the treatments the most recommended for cryopreservation of tambaqui semen.(AU)

Avaliação de diferentes crioprotetores e tempos de descongelamento de sêmen de tambaqui Colossoma macropomum / Evaluation of different cryoprotectants and thawing times of tambaqui Colossoma macropomum semen

O objetivo com o trabalho foi avaliar a eficiência de quatro crioprotetores adicionados ao meio diluidor na criopreservação do sêmen de tambaqui (Colossoma macropomum) e melhor protocolo de descongelamento. Foram realizadas quatro coletas com intervalos regulares de amostras de sêmen (pool) de dez reprodutores. Os crioprotetores testados foram: glicerol, dimeltilsulfóxido (DMSO), dimetilformamida (DMF) e metanol, na proporção de 10,0%. Para a avaliação da qualidade do sêmen após o descongelamento foi realizado o teste in vivo das amostras em três protocolos diferentes para cada crioprotetor, com três tempos de descongelamento (15, 30 e 45s) na temperatura de 30ºC, totalizando 12 tratamentos com cinco repetições em esquema fatorial. Os resultados da fertilização foram submetidos a análise de variância (ANOVA) e teste de Tukey, com significância de 0,05. O tratamento que resultou em melhor taxa de fertilização foi o que continha metanol como crioprotetor e o descongelamento no tempo de 30 s, com taxa de fertilização média de 77,65%, sendo, entre os tratamentos avaliados, o mais recomendado para a criopreservação do sêmen de tambaqui.

The objective of the study was to evaluate the efficiency of four cryoprotectants added to the extender on sperm cryopreservation of tambaqui Colossoma macropomum and to define the best thawing protocol. Four semen samples were collected at regular intervals from ten breeding males (pool). The cryoprotectants tested were: glycerol, dimethylsulfoxide (DMSO), dimethylformamide (DMF) and methanol at a ratio of 10.0%. For the evaluation of semen quality after thawing in vivo test samples in three different protocols for each cryoprotectant was performed with three thawing times (15, 30 and 45s) at 30ºC temperature, totalizing 12 treatments with five replicates in factorial design. The results of fertilization were subjected to analysis of variance (ANOVA) and Tukey's test, with significance of 0.05. The treatment that resulted in improved fertilization rate was the one containing methanol as cryoprotectant and thawing time of 30 s, with a mean fertilization rate of 77.65%, being among the treatments the most recommended for cryopreservation of tambaqui semen.

Protein profile as a quality indicator of cryopreserved semen from Tambaqui Colossoma macropomum (Cuvier, 1818)

Abstract The aim of this study was to evaluate the association between proteins in the seminal plasma of tambaqui Colossoma macropomum (Cuvier, 1818) with seminal quality indicators after thawing. The semen was cryopreserved with a dilution based on BTS with 8% DMSO. A 200 µL sample of semen from each animal was diluted in 800 µL BTS, centrifuged at 800 rpm, and the supernatant was cryopreserved to further analyze of the protein profile of seminal plasma through one-dimensional electrophoresis (SDS-PAGE). After 15 days of cryopreservation, a cryopreserved semen straw was thawed to analyze both qualitative and quantitative parameters. When considering all collections, the SDS-PAGE identified 15 protein bands in the seminal plasma of tambaqui. When the interaction (presence or absence) between proteins observed in the seminal plasma and the post thawed spermatic parameters was evaluated, we observed a great influence of the presence of proteins on spermatic quality. A greater (P 0.05) fertilization rate was observed with the presence of proteins 12, 34, 44, 85, and 90 kDa. Proteins in seminal plasma of tambaqui influenced the spermatic quality after thawing, and thus, they can be utilized as an indicator of sperm quality, especially the proteins with a molecular weight 50 kDa.

Resumo O objetivo desse estudo foi de avaliar a associação entre a presença de proteínas no plasma seminal do tambaqui Colossoma macropomum (Cuvier, 1818) com indicadores de qualidade seminal pós-descongelamento. O semen foi criopreservado com diluidor a base de BTS com 8% DMSO. Uma amostra de 200 µL de semen de cada animal foi diluída em 800 µL de BTS, e centrifugada em 800 rpm, e somente o sobrenadante foi criopreservado para posterior análise do perfil proteico do plasma seminal, através da eletroforese unidimensional (SDS-PAGE). Decorridos 15 dias da criopreservação, uma palheta com semen criopreservado foi descongelado para análise dos parâmetros quali-quantitativos. Considerando todas as coletas, o SDS-PAGE identificou 15 bandas proteicas no plasma seminal do tambaqui. Quando se avaliou a interação (presença ou ausência) das proteínas encontradas no plasma seminal, com os parâmetros espermáticos pós-descongelamento, observou-se grande influência da presença das proteínas na qualidade espermática. Observou-se maior taxa de fertilização (P 0,05) com a presença das proteínas 12, 34, 44, 85 e 90 kDa. As proteínas do plasma seminal de tambaqui influenciaram na qualidade espermática após descongelamento, podendo ser utilizadas como indicadores para a qualidade espermática após descongelamento, principalmente as proteínas com peso molecular 50 kDa.

Oocyte quality of tambaqui (Colossoma macropomum) during the reproductive season / Qualidade ovocitária de tambaqui (Colossoma macropomum) ao longo da estação reprodutiva

The study aimed to analyze the Colossoma macropomum reproductive behavior and quality of the female gametes throughout the reproductive season. The experiment was carried out in Pimenta Bueno - Rondônia State (Northern Brazil) during the reproductive season (2010-2011) using 36 females. Each sampling was performed on a 15 ± 5 days interval. Female gametes were collected by stripping and the following analyses were performed: weight of oocytes released (g); productivity index, fertilization and hatching rate. During the sampling period was verified effect (p < 0.05) of collecting time into the season for oocytes weight, productivity index and fertilization rate. Although the period 3 (December) did not differ significantly from other periods, it showed better parameters for the quality of C. macropomum oocytes.(AU)

O estudo foi conduzido com o objetivo de analisar o comportamento reprodutivo da espécie Colossoma macropomum, quanto à qualidade de seus gametas femininos ao longo da estação reprodutiva. O experimento foi executado em Pimenta Bueno-Rondônia durante a estação reprodutiva do C. macropomum. Utilizaram 36 fêmeas durante a estação de 2010-2011. Cada coleta apresentou um intervalo de 15±5 dias. Através da extrusão foram coletados os gametas femininos e realizadas as seguintes análises ao longo da estação: peso de oócitos liberados (g); índice de produtividade; taxa de fertilização e eclosão. Durante a estação 2010-2011 foi verificado efeito (p < 0,05) de período (coleta) dentro da estação para peso de oócitos, índice de produtividade e taxa de fertilização. Apesar do período 3 (coleta mês de dezembro) não ter diferenciado significativamente de alguns períodos, foi o que apresentou os melhores parâmetros estabelecidos para a qualidade dos oócitos de C. macropomum.(AU)

Oocyte quality of tambaqui (Colossoma macropomum) during the reproductive season / Qualidade ovocitária de tambaqui (Colossoma macropomum) ao longo da estação reprodutiva

The study aimed to analyze the Colossoma macropomum reproductive behavior and quality of the female gametes throughout the reproductive season. The experiment was carried out in Pimenta Bueno - Rondônia State (Northern Brazil) during the reproductive season (2010-2011) using 36 females. Each sampling was performed on a 15 ± 5 days interval. Female gametes were collected by stripping and the following analyses were performed: weight of oocytes released (g); productivity index, fertilization and hatching rate. During the sampling period was verified effect (p < 0.05) of collecting time into the season for oocytes weight, productivity index and fertilization rate. Although the period 3 (December) did not differ significantly from other periods, it showed better parameters for the quality of C. macropomum oocytes.

O estudo foi conduzido com o objetivo de analisar o comportamento reprodutivo da espécie Colossoma macropomum, quanto à qualidade de seus gametas femininos ao longo da estação reprodutiva. O experimento foi executado em Pimenta Bueno-Rondônia durante a estação reprodutiva do C. macropomum. Utilizaram 36 fêmeas durante a estação de 2010-2011. Cada coleta apresentou um intervalo de 15±5 dias. Através da extrusão foram coletados os gametas femininos e realizadas as seguintes análises ao longo da estação: peso de oócitos liberados (g); índice de produtividade; taxa de fertilização e eclosão. Durante a estação 2010-2011 foi verificado efeito (p < 0,05) de período (coleta) dentro da estação para peso de oócitos, índice de produtividade e taxa de fertilização. Apesar do período 3 (coleta – mês de dezembro) não ter diferenciado significativamente de alguns períodos, foi o que apresentou os melhores parâmetros estabelecidos para a qualidade dos oócitos de C. macropomum.