ResumoO objetivo deste estudo foi avaliar os efeitos da congelação sobre a integridade e funcionalidade de espermatozoides colhidos do epidídimo de touros post-mortem. Foram utilizados 10 pares de testículos de touros provenientes de um abatedouro comercial. No laboratório, o sêmen foi colhido da cauda do epidídimo sobre uma placa de Petri aquecida, contendo meio diluente sem glicerol (Botubov® -meio I, Botupharma, Botucatu, SP). O sêmen foi então avaliado quanto à motilidade, vigor, concentração, morfologia espermática, integridade de DNA, atividade mitocondrial, viabilidade, integridade e funcionalidade da membrana. Após a avaliação, o sêmen foi diluído em meio comercial contendo glicerol (Botubov® -meio II, Botupharma, Botucatu, SP), em uma concentração de 100x10 6 de espermatozoides/mL. Ato contínuo, as amostras foram envasadas em palhetas de 0,5 mL, refrigeradas a 4ºC, durante 4 horas, congeladas em caixa de poliestireno expandido, a 6cm do nível de nitrogênio líquido, durante 20 minutos e, posteriormente, armazenada em botijão criogênico. As amostras foram descongeladas a 37°C, durante 30 segundos. Após a descongelação, o sêmen foi reavaliado para os mesmos parâmetros, com exceção da concentração espermática e volume. Quanto à comparação dos resultados entre o sêmen fresco e congelado, estes foram superiores (p<0,05) no sêmen fresco quanto à motilidade (74,0 ± 7,0% versus 30,0 ± 8,2%), vigor (3,6 ± 0,7 versus 1,3 ± 0,5), integridade de membrana (fluorescência, 95,6 ± 3,1% versus 75,7 ± 5,9%) e índice de atividade citoquímica mitocondrial (99,3 ± 6,6 versus 40,0 ± 6,3), respectivamente. Concluiuse que a viabilidade do sêmen congelado obtido do epidídimo de touros é semelhante à do sêmen ejaculado, tornando a técnica de colheita aplicável comercialmente.Palavras-chave: criopreservação; ejaculado; bovino; espermatozoide; epididimal. AbstractThe objective of this study was to evaluate the effects of freezing on the integrity and functionality of bovine epididymal spermatozoa collected postmortem. Ten pairs of testicle epididymis of bulls from a commercial slaughterhouse were used. On the laboratory, the testicle was separated and the semen was collected from the tail of epididymis on a petri dish heated containing a medium (Botubov ®, Botupharma, Botucatu, SP). Then, the semen was evaluated to motility, vigor, concentration, morphology, DNA integrity, mitochondrial activity, viability, membrane integrity and functionality. The semen was diluted in commercial extender (Botubov ®, Botupharma, Botucatu, SP) at a concentration of 100x10 6 sperm/mL. Thereafter, the samples were packaged in straws 0.5 mL, chilled at 4° C, for 4 hours, frozen in polystyrene box, at 6 cm from level of liquid nitrogen, during 20 minutes and stored in cryogenic container. The samples were thawed at 37°C for 30 seconds. After thawing, the semen was reevaluated to the same parameters, except sperm concentration and volume. Comparison of results between fresh and frozen semen showed means were increased (p<0.05) fresh semen motility (74,0 ...

The objective of this study was to evaluate the conception rate of crossbred heifers (n=50) and cows (n=50) inseminated with sexed and conventional semen between 18 and 24 hours after estrous detection. The synchronization protocol of the estrous cycle started on day zero (D0) by inserting the intravaginal device with 1g progesterone (Sincrogest® Ourofino, Brazil) and injecting 2 mg of estradiol benzoate, intramuscularly (Sincrodiol® Ourofino, Brazil). On the fifth day (D5), 200 IU of equine chorionic gonadotrophin was injected intramuscularly (Folligon®, Intervet, Brazil). On the eighth day (D8), after removing the progesterone device, 500 g of sodium cloprostenol was injected intramuscularly (Sincrocio®, Ourofino, Brazil). After that, the animals were checked for estrus 3 times daily, and inseminated 18 to 24 hours after estrus detection. Pregnancy diagnosis was performed 30 to 40 days after insemination. Conception rate did not differ (P> 0.05) according to animal category, but was higher for conventional semen compared to sexed semen when evaluating the total of animals and lactating cows (P <0.05). Artificial insemination of heifers with sexed semen 18 to 24 hours after estrus detection was effective, however, conventional semen was more efficient in lactating cows.Keywords: bovine, spermatozoids, heat. ResumoConsiderando os benefícios do uso de sêmen sexado e também os danos causados pelo processo de separação dos espermatozoides, o objetivo do presente estudo foi avaliar a taxa de concepção de novilhas (n=50) e vacas (n=50) mestiças inseminadas com sêmen sexado e convencional após 18 a 24 horas a observação do cio. O protocolo de sincronização do ciclo estral consistiu em inserção de dispositivo intravaginal com 1g de progesterona (Sincrogest® Ourofino, Brasil) e aplicação intramuscular de 2mg de benzoato de estradiol (Sincrodiol® Ourofino, Brasil) no dia zero (D0). No quinto dia (D5), foi realizada uma aplicação intramuscular de 200UI de gonadotrofina coriônica equina (Folligon®, Intervet, Brasil). No oitavo dia (D8), o dispositivo de progesterona foi retirado, e aplicado por via intramuscular 500µg de cloprostenol sódico (Sincrocio®, Ourofino, Brasil). A partir deste momento, o estro foi observado 3 vezes ao dia e os animais foram inseminados 18 a 24 após a detecção do cio. O diagnóstico de gestação foi realizado 30 a 40 dias após a inseminação. Não foi observada diferença na taxa de concepção de acordo com a categoria animal (P > 0,05), entretanto, animais inseminados com sêmen convencional apresentaram melhor taxa de concepção do que com sêmen sexado quando se avaliou o total de animais e vacas lactantes (P < 0,05). A inseminação artificial de novilhas com sêmen sexado 18 a 24 horas após detecção de estro mostrou-se eficaz, entretanto, para vacas lactantes não foi observada a mesma eficiência ao se comparar com o sêmen convencional.

The reproductive biotechnologies directly assist in the development of different methods of reproductive evaluation. The objective of this study was to develop a new method to determine the estrous cycle phase in equine females through vaginal cytology throughout the seasons, as well as to establish an estimate of inflammatory cells present in the vagina. Six mares were evaluated at different ages without a defined breed and reproductive activity and were subjected to ultrasound evaluation of the reproductive system and cytological analysis of the vaginal region. In the summer, there was a predominance of keratinized epithelial cells in the estrus and intermediate in the diestrus. In the autumn, there was a predominance of keratinized cells in estrus and parabasal cells in estrus and diestrus. In anestrus (winter), a greater number of parabasal cells was identified in relation to the other cell types. In estrus (spring), there was a predominance of parabasal and intermediate cells. Conversely, in diestrus, parabasals were found in greater numbers. The evaluation of inflammatory cells of the vaginal epithelium of mares showed greater activity in the summer; however, there are no reference values for healthy animals in the literature, and it is necessary to conduct studies on the subject. It is concluded that there is an influence of cyclicity on the vaginal epithelium of the equine species, varying according to the season. Additionally, vaginal cytological evaluation is an important complementary tool in the diagnosis of vaginitis in mares, requiring further research on the subject.