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The objective of this study was to evaluate the effect of retinol (RT) and retinoic acid (RA) on the in vitro development of pre-implantation goat embryos cultured in potassium simplex optimized medium or synthetic oviduct fluid or cocultured in oviductal cells monolayer either in potassium simplex optimized medium or synthetic oviduct fluid. A total of 2407 cumulus-oocyte complexes were aspirated from 2 to 6 mm ovarian follicles from slaughtered animals. Selected cumulus-oocyte complexes were subjected to in vitro maturation in TCM 199 for 24 h at 39 °C in an atmosphere of 5% (v/v) CO(2) in humidified air. In vitro fertilization was performed in modified defined medium. Eighteen hours after in vitro fertilization, cumulus cells were removed and presumptive zygotes were randomly distributed into experimental groups. In Experiment 1, presumptive zygotes were cultured in potassium simplex optimized medium, potassium simplex optimized medium + RT, potassium simplex optimized medium + retinoic acid, synthetic oviduct fluid, synthetic oviduct fluid + RT and synthetic oviduct fluid + RA at 39 °C in a humidified atmosphere of 5% (v/v) CO(2), 5% (v/v) O(2) and 90% (v/v) N(2). In Experiment 2, presumptive zygotes were cocultured in potassium simplex optimized medium + oviductal cells monolayer, potassium simplex optimized medium + RT + oviductal cells monolayer, potassium simplex optimized medium + RA + oviductal cells monolayer, synthetic oviduct fluid + oviductal cells monolayer, synthetic oviduct fluid + RT + oviductal cells monolayer and synthetic oviduct fluid + RA + oviductal cells monolayer in an atmosphere of 5% (v/v) CO(2) in humidified air. In both experiments, media were partially changed on day 2 after in vitro fertilization and unfertilized oocytes were excluded from the experiment. Embryos were cultured or cocultured for 8 days. In Experiment 1, there was no effect of RT or RA supplementation on the proportion of oocytes that reached the morula or blastocyst stages. By contrast, Experiment 2 demonstrated that the addition of 0.28 μg/ml RT and 0.5 μm RA to the embryo culture media stimulated (p < 0.05) development to the morula and blastocyst stages under the coculture conditions tested. In conclusion, retinoids play an important role in pre-implantation development of goat embryos and can be used to enhance in vitro embryo production.

ContentsThe objective of this study was to evaluate the effect of retinol on the in vitro development of early embryos of cultured Bos indicus (Expt 1) to the blastocyst stage in medium simplex of optimization (KSOM) or sintetic fluid of oviduct (SOF) or co-cultured (Expt 2) with an oviduct cell monolayer (OCM) in KSOM or SOF. A total of 3149 cumulus-oocyte complexes obtained by aspirating follicles (2-5 mm diameter) from ovaries of slaughtered animals were selected for IVM and incubated in TCM 199 supplemented with 25 mM HEPES at 39°C in air with 5% CO 2 and maximum humidity for 24 h. In vitro fertilization (IVF) was performed in modified defined medium (mDM) medium. Eighteen hours after IVF, cumulus cells were removed and presumptive zygotes were randomly allocated to the experimental groups. Zygotes cultured (Expt 1) in KSOM + retinol, KSOM, SOF + retinol and SOF were incubated in maximum humidity at 39°C, 5% CO 2 , 5% O 2 and 90% N 2 . Zygotes co-cultured (Expt 2) in KSOM + retinol + OCM, KSOM + OCM, SOF + retinol + OCM and SOF + OCM were incubated at 39°C, 5% CO 2 . In both experiments media were partially changed 48 h after IVF and unfertilized ova were removed. Afterwards embryos were kept in culture or co-culture for further 9 days. In Expt 1, blastocyst rates (day 7) were 14.6% (KSOM + retinol), 15.8% (KSOM), 16.4% (SOF + retinol) and 15.9% (SOF). In Expt 2, the blastocyst rates (day 7) were 25.4% (KSOM + retinol + OCM) 14.2% (KSOM + OCM), 24.3% (SOF + retinol + OCM) and 15.9% (SOF + OCM). The same influence profile of retinol was observed in the formation of the expanded (day 9) and hatched (day 11) blastocysts. The results obtained in Expt 2 demonstrated that the addition of 0.28 lg/ml retinol to the embryo culture media used in this study had a significant (p < 0.05) positive effect on bovine early embryonic development, under the conditions tested, and can be used to enhance in vitro embryo production.

The aim of this study was to determine the period of genital tubercle (GT) migration using ultrasonography in Morada Nova sheep foetuses (n = 117) from natural mating (NM) and frozen embryo transfer (ET) to determine the window when foetal sexing can be determined. The examinations were performed using transrectal ultrasonography with a dual-frequency linear transducer (6.0 and 8.0 MHz) from day 30-54 of pregnancy at 48-h intervals. The period of GT migration of foetuses produced by NM varied from 36 to 46 days of pregnancy, resulting in an average of 39.5 +/- 2.9 days. For foetuses derived from ET, GT migration varied from 42 to 52 days of pregnancy with an average of 48.5 +/- 3.3 days, being possible the GT of foetuses from ET start to migrate 96 h later even if they are of the same gender. Migration of the GT occurred earlier (p < 0.05) in foetuses produced by NM and sexing accuracy for triplet pregnancies (77.8%) was significantly inferior (p < 0.05) to single (100%) and twin (92.9%) pregnancies for foetuses derived by NM. The results allow one to conclude that foetal sexing can be done from the 50th day onwards in foetuses produced by NM and from the 55th day onwards in foetuses derived from ET, and that multiple pregnancies compromise the sexing accuracy by ultrasonography.

This study aimed to evaluate the effect of temporary suckling interruption associated with the male effect on reproductive performance of pluriparous Santa Inês ewes. The females were kept apart from the males for 60 days and then randomly distributed into three treatments associated with the male effect (DT0, DT24 and DT48); in DT0, there was no suckling interruption; in DT24, suckling was interrupted for 24 hours, and in DT48, sucking was interrupted for 48 hours. Estrous distribution was observed within 31 (DT0), 27 (DT24) and 38 (DT48) days of the breeding season. Estrous synchronization up to the 5th day of the mating season was observed in 15% (DT0), 30% (DT24) and 25% (DT48) of the females, with no difference among treatments. Estrous percentages were 90% (DT0), 100% (DT24) and 100% (DT48), with no difference among treatments. Pregnancy rates were 38.4% (DT0), 60.0% (DT24) and 45.0% (DT48) with no difference among treatments. Prolificacy was 1.43 (DT0), 1.17 (DT24) and 1.22 (DT48) and did not differ between treatments. In conclusion, temporary suckling interruption associated with the male effect is efficient to induce estrous but not to synchronize estrous or increase the pregnancy rates and prolificacy of Santa Inês ewes during a 45-day breeding season.Keywords: anestrous, biostimulation, reproduction, sheep. EFEITO MACHO ASSOCIADO AO DESMAME TEMPORÁRIO NO DESEMPENHO REPRODUTIVO DE OVELHAS SANTA INÊSRESUMO: O estudo teve como objetivo avaliar o efeito do desmame temporário associado ao efeito macho sobre o desempenho reprodutivo de ovinos Santa Inês. As fêmeas foram mantidas distantes dos machos por 60 dias e aleatoriamente distribuídas em três tratamentos associados ao efeito macho (DT0, DT24 e DT48), no qual em DT0, não houve interrupção da amamentação; em DT24, amamentação interrompida por 24 horas e em DT48, interrupção da amamentação por 48 horas. Distribuição de estro foi observada em 31 (DT0), 27 (DT24) e 38 (DT48) dias da estação de monta. Sincronização de estro até o quinto dia da estação de monta foi observada em 15% (DT0), 30% (DT24) e 25% (DT48) das fêmeas, não havendo diferença entre os tratamentos. Percentagens de estro foram de 90% (DT0), 100% (DT24) e 100% (DT48), não havendo diferença entre os tratamentos. As taxas de prenhez foram de 38,4% (DT0), 60,0% (DT24) e 45,0% (DT48), sem diferença entre os tratamentos. A prolificidade foi de 1,43 (DT0), 1,17 (DT24) e 1,22 (DT48), e não diferiu entre os tratamentos. Em conclusão, o desmame temporário associado ao efeito macho é eficiente na indução do estro, embora não seja eficiente na sincronização de estros e não aumente as taxas de gestação e prolificidade das ovelhas Santa Inês durante uma estação de monta de 45 dias.Palavras-chave: anestro, bioestimulação, ovinos, reprodução.