Effect of different gonadotropins on in vitro maturation of sheep oocytes / Efeito de diferentes gonadotrofinas na maturação in vitro de oócitos ovinos

The aim of this study was to examine the effect of replacing the use of follicle-stimulating hormone (FSH) with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) on the in vitro maturation (IVM) of sheep oocytes. After sheep ovaries were collected (n=300), the cumulus-oocyte complexes were aspirated, selected, and divided into four groups according to the IVM medium: CON group, in which the basic IVM medium was used; and eCG, hCG, and FSH groups, in which the oocytes were immersed in basic IVM medium with 10 IU/mL eCG, 10 IU/mL hCG, and 10 µg/mL FSH-p, respectively. In vitro maturation of the oocytes was performed at 38.5 °C, in a humidified atmosphere of 5% CO2 in air, for 24 h. Subsequently, the oocytes were evaluated for the degree of cumulus-cell expansion, chromatin configuration, GSH levels, and active mitochondria. There were no significant differences for the rate of cumulus cell expansion. The percentage of oocytes in MII was higher in the eCG group than in the CON and hCG groups (P<0.05) and similar to that of the FSH group. In conclusion, eCG can be used as a substitute for FSH in IVM of sheep oocytes.

O objetivo deste estudo foi avaliar o efeito da gonadotrofina coriônica equina (eCG) e da gonadotrofina coriônica humana (hCG), em substituição ao uso de hormônio folículo estimulante (FSH) na maturação in vitro (MIV) de oócitos ovinos. Após a coleta de ovários (n=300) ovinos, os complexos cúmulus-oócitos (CCOs) foram aspirados, selecionados e divididos em quatro grupos de acordo com o meio de MIV: grupo CON, em que foi utilizado o meio MIV base; e grupos ECG, HCG e FSH, em que os oócitos foram imersos em meio MIV base adicionado de 10 UI/mL de eCG, 10 UI/mL de hCG e 10 µg/mL de FSH-p, respectivamente. A MIV dos oócitos foi realizada a 38,5°C, em atmosfera umidificada de 5% de CO2 em ar, durante 24 horas. Posteriormente, os oócitos foram avaliados, quanto grau de expansão das células do cumulus, configuração da cromatina, níveis de GSH e mitocôndrias ativas. Não foram observadas diferenças significativas com relação à taxa de expansão de células do cumulus. A percentagem de oócitos em MII foi maior no grupo ECG do que no grupo CON e HCG (P<0,05) e semelhante ao grupo FSH. Em conclusão, a eCG pode ser utilizada em substituição ao FSH na MIV de oócitos ovinos.

Supplemented powdered coconut water (ACP-406®) promotes growth of goat secondary follicles and oocyte meiotic resumption

The objective of this study was to test the efficiency of powdered coconut water (ACP-406®) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406®, both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM+ and ACP+). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM+, ACP and ACP+, and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP+ than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP+ and α-MEM+ treatments. Both ACP+ and α-MEM+ allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days.(AU)

Supplemented powdered coconut water (ACP-406®) promotes growth of goat secondary follicles and oocyte meiotic resumption

The objective of this study was to test the efficiency of powdered coconut water (ACP-406®) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406®, both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM+ and ACP+). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM+, ACP and ACP+, and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP+ than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP+ and α-MEM+ treatments. Both ACP+ and α-MEM+ allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days.

Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo / Adição de sacarose na solução crioprotetora de vitrificação melhora a qualidade de embriões ovinos Dorper produzidos in vivo

This study aimed to evaluate the efficacy of adding sucrose in vitrification solution of ovine embryos produced in vivo. Forty Dorper ewes were selected and superovulated. Immediately prior to the embryo collection by laparotomy, a laparoscopy was performed to verify the superovulatory response. The recovered flushing was followed by embryo evaluation and embryos were divided in two experimental groups where embryos from Control group were submitted to a traditional vitrification protocol and embryos from Sucrose group to a modified vitrification protocol with sucrose. After warming, embryos were again divided regarding cryoprotectant removal (Indirect) or not (Direct). The embryo quality was classified as embryos of degrees I (excellent or good), II (regular), III (poor) and IV (dead or degenerate). It was also verified the homogeneity of mass, occurrence of embryonic mass retraction and rupture of pellucid zone. The results were expressed as percentages and were subjected to Chi-square test with P 0.05. The embryos vitrified in the presence of sucrose had lower proportions of lower-quality embryos after warming (22.20 vs. 44.50%), higher percentages of homogeneous embryos after warming (63.89 vs. 38.89 %) while concerning other parameters there was no difference between these groups. It can be concluded that the addition of 0.4 M sucrose during vitrification improves the embryo quality.(AU)

Este estudo objetivou avaliar a eficácia da adição de sacarose na solução de vitrificação de embriões ovinos produzidos in vivo. Foram selecionadas 40 ovelhas da raça Dorper as quais foram superovuladas. Imediatamente antes da colheita de embriões por laparotomia, uma laparoscopia foi realizada para verificar a resposta superovulatória. O lavado recuperado foi submetido à procura e avaliação de embriões e estes foram divididos em dois grupos experimentais, onde os embriões do Grupo Controle foram submetidos ao protocolo tradicional de vitrificação e os embriões do Grupo Sacarose a um protocolo modificado de vitrificação com sacarose. Após a descongelação, os embriões foram novamente divididos considerando a remoção (Indireto) ou não (Direto) do crioprotetor. A qualidade embrionária foi classificada como embriões de graus I (excelente ou bom), II (regular), III (pobre) e IV (morto ou degenerado). Foi também verificada a homogeneidade da massa, ocorrência de retração da massa e ruptura de zona pelúcida. Os resultados foram expressos em porcentagem e foram submetidos ao teste do Qui-quadrado com P < 0.05. Os embriões vitrificados na presença de sacarose apresentaram menores proporções de embriões de menor qualidade após a descongelação (22,20 vs. 44,50%), e maiores percentuais de embriões homogêneos após a descongelação (63,89 vs. 38,89%) enquanto com relação aos demais parâmetros não existiram diferenças entre grupos. Pode-se concluir que a adição de 0,4 M de sacarose durante os procedimentos de vitrificação e descongelação beneficia a qualidade embrionária.(AU)

Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo / Adição de sacarose na solução crioprotetora de vitrificação melhora a qualidade de embriões ovinos Dorper produzidos in vivo

This study aimed to evaluate the efficacy of adding sucrose in vitrification solution of ovine embryos produced in vivo. Forty Dorper ewes were selected and superovulated. Immediately prior to the embryo collection by laparotomy, a laparoscopy was performed to verify the superovulatory response. The recovered flushing was followed by embryo evaluation and embryos were divided in two experimental groups where embryos from Control group were submitted to a traditional vitrification protocol and embryos from Sucrose group to a modified vitrification protocol with sucrose. After warming, embryos were again divided regarding cryoprotectant removal (Indirect) or not (Direct). The embryo quality was classified as embryos of degrees I (excellent or good), II (regular), III (poor) and IV (dead or degenerate). It was also verified the homogeneity of mass, occurrence of embryonic mass retraction and rupture of pellucid zone. The results were expressed as percentages and were subjected to Chi-square test with P 0.05. The embryos vitrified in the presence of sucrose had lower proportions of lower-quality embryos after warming (22.20 vs. 44.50%), higher percentages of homogeneous embryos after warming (63.89 vs. 38.89 %) while concerning other parameters there was no difference between these groups. It can be concluded that the addition of 0.4 M sucrose during vitrification improves the embryo quality.

Este estudo objetivou avaliar a eficácia da adição de sacarose na solução de vitrificação de embriões ovinos produzidos in vivo. Foram selecionadas 40 ovelhas da raça Dorper as quais foram superovuladas. Imediatamente antes da colheita de embriões por laparotomia, uma laparoscopia foi realizada para verificar a resposta superovulatória. O lavado recuperado foi submetido à procura e avaliação de embriões e estes foram divididos em dois grupos experimentais, onde os embriões do Grupo Controle foram submetidos ao protocolo tradicional de vitrificação e os embriões do Grupo Sacarose a um protocolo modificado de vitrificação com sacarose. Após a descongelação, os embriões foram novamente divididos considerando a remoção (Indireto) ou não (Direto) do crioprotetor. A qualidade embrionária foi classificada como embriões de graus I (excelente ou bom), II (regular), III (pobre) e IV (morto ou degenerado). Foi também verificada a homogeneidade da massa, ocorrência de retração da massa e ruptura de zona pelúcida. Os resultados foram expressos em porcentagem e foram submetidos ao teste do Qui-quadrado com P < 0.05. Os embriões vitrificados na presença de sacarose apresentaram menores proporções de embriões de menor qualidade após a descongelação (22,20 vs. 44,50%), e maiores percentuais de embriões homogêneos após a descongelação (63,89 vs. 38,89%) enquanto com relação aos demais parâmetros não existiram diferenças entre grupos. Pode-se concluir que a adição de 0,4 M de sacarose durante os procedimentos de vitrificação e descongelação beneficia a qualidade embrionária.

Using the same CIDR up to three times for estrus synchronization and artificial insemination in dairy goats / Uso do mesmo CIDR por até três vezes para sincronização do estro e inseminação artificial de cabras leiteiras

The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 +-g d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 +- 1.1h vs. 13.8 +- 2.6h vs. 13.3 +-1.4h), as well as estrus duration (33.6 +- 7.3h vs. 29.6 +-3.2h vs. 32.8 +- 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.

Objetivou-se avaliar o efeito da utilização do mesmo dispositivo de liberação controlada de drogas (CIDR) por até três vezes sobre o desempenho reprodutivo de cabras leiterias exploradas no semiárido do Nordeste Brasileiro. Foram utilizadas 45 cabras divididas em três tratamentos de sincronização do estro, sendo: CIDR1x, tratadas com CIDR novo durante nove dias. Dois dias antes da retirada do dispositivo, foram aplicados 75 +-g de d-cloprostenol e 300 UI de gonadotrofina coriônica eqüina (eCG). Para os demais tratamentos, foi utilizado o mesmo protocolo hormonal, diferindo apenas pelo uso do mesmo CIDR pela segunda vez no grupo CIDR2x e uso pela terceira vez no grupo CIDR3x. O intervalo entre a retirada do dispositivo e o início do estro (13,3+- 1,1h vs. 13,8 +- 2,6h vs. 13,3 +- 1,4h), bem como, a duração do estro (33,6 +- 7,3h vs. 29,6 +- 3,2h vs. 32,8 +- 4,5h) não diferiram (p > 0,05) entre os grupos CIDR1x, CIDR2x e CIDR3x, respectivamente. Todas as fêmeas sincronizadas foram identificadas em estro. As médias de fertilidade e prolificidade média após inseminação artificial foram, respectivamente, de 82,2% e 1,9 crias, não havendo diferença significativa (p > 0,05) entre os tratamentos. A utilização do mesmo CIDR por até três vezes foi viável na sincronização do estro de caprinos leiteiros.

Using the same CIDR up to three times for estrus synchronization and artificial insemination in dairy goats - doi: 10.4025/actascianimsci.v33i3.10120 / Using the same CIDR up to three times for estrus synchronization and artificial insemination in dairy goats - doi: 10.4025/actascianimsci.v33i3.10120

The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.

The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.