RESUMO

Objetivou-se investigar a eficiência dos meios TCM 199 e PBS na conservação de folículos pré-antrais(FOPA) felinos in situ a 4ºC por 24 h. Foram utilizados 10 pares de ovários de gatas Felis catus, sendo a camadacortical fracionada e submetida a três diferentes tratamentos: controle, T1 (fragmentos conservados em TCM199) e T2 (fragmentos conservados em PBS). Realizou-se a preparação histológica do grupo controle, sendo T1e T2 mantidos a 4ºC, por 24 h, em caixa isotérmica. Ao final dos tratamentos, os FOPA foram classificados emprimordiais, primários e secundários, e os oócitos classificados em normais ou degenerados morfologicamente. Aavaliação histológica constatou que os FOPA do controle (63%) tiveram maior integridade morfológica (P > 0,05)quando comparados ao T2 (55%), sendo semelhantes ao T1 (59%). Não houve diferença (P < 0,05) entreT1(59%) e T2 (55%). Os folículos primordiais mantiveram maior integridade morfológica quando comparadosaos folículos em desenvolvimento, independentemente do tratamento. Com base nesses resultados, pode-seconcluir que é possível a manutenção da integridade morfológica de FOPA em ovários de gatas mantidos a 4ºCpor 24 h, utilizando o TCM 199 e que os folículos primordiais têm maior viabilidade, quando comparados aosfolículos em desenvolvimento.(AU)

The aim of this study was to investigate the efficiency of TCM 199 and PBS mediums in the conservationof feline preantral follicles (PAF) in situ at 4°C for 24 h. Ten pairs of ovaries of Felis catus cats were used, inwhich the cortical layer was fractioned and submitted to three different treatments: Control, T1 (fragmentspreserved in TCM 199) and T2 (fragments preserved in PBS). Histological preparation was carried out in thecontrol group, while T1 and T2 were kept at 4°C for 24 h in an isothermal box. After histological preparation ofControl, T1 and T2 the PAF were classified as primordial, primary and secondary and the morphology of theoocyte as normal and degenerated. Histological evaluation found that the PAF control group (63%) had a highermorphological integrity (P > 0.05) compared to T2 (55%), which was similar to T1 (59%). However, there wasno difference (P < 0.05) between T1 (59%) and T2 (55%) treatments. Primordial follicles maintained greatermorphological integrity when compared to developing follicles, regardless of the treatment. Based on theseresults, it can be concluded that it is possible to maintain the morphological integrity of PAF in ovaries of catstransported at 4°C for 24 h using TCM 199 and that primordial follicles have a higher viability compared todeveloping follicles.(AU)

Assuntos

RESUMO

Objetivou-se investigar a eficiência dos meios TCM 199 e PBS na conservação de folículos pré-antrais(FOPA) felinos in situ a 4ºC por 24 h. Foram utilizados 10 pares de ovários de gatas Felis catus, sendo a camadacortical fracionada e submetida a três diferentes tratamentos: controle, T1 (fragmentos conservados em TCM199) e T2 (fragmentos conservados em PBS). Realizou-se a preparação histológica do grupo controle, sendo T1e T2 mantidos a 4ºC, por 24 h, em caixa isotérmica. Ao final dos tratamentos, os FOPA foram classificados emprimordiais, primários e secundários, e os oócitos classificados em normais ou degenerados morfologicamente. Aavaliação histológica constatou que os FOPA do controle (63%) tiveram maior integridade morfológica (P > 0,05)quando comparados ao T2 (55%), sendo semelhantes ao T1 (59%). Não houve diferença (P < 0,05) entreT1(59%) e T2 (55%). Os folículos primordiais mantiveram maior integridade morfológica quando comparadosaos folículos em desenvolvimento, independentemente do tratamento. Com base nesses resultados, pode-seconcluir que é possível a manutenção da integridade morfológica de FOPA em ovários de gatas mantidos a 4ºCpor 24 h, utilizando o TCM 199 e que os folículos primordiais têm maior viabilidade, quando comparados aosfolículos em desenvolvimento.

The aim of this study was to investigate the efficiency of TCM 199 and PBS mediums in the conservationof feline preantral follicles (PAF) in situ at 4°C for 24 h. Ten pairs of ovaries of Felis catus cats were used, inwhich the cortical layer was fractioned and submitted to three different treatments: Control, T1 (fragmentspreserved in TCM 199) and T2 (fragments preserved in PBS). Histological preparation was carried out in thecontrol group, while T1 and T2 were kept at 4°C for 24 h in an isothermal box. After histological preparation ofControl, T1 and T2 the PAF were classified as primordial, primary and secondary and the morphology of theoocyte as normal and degenerated. Histological evaluation found that the PAF control group (63%) had a highermorphological integrity (P > 0.05) compared to T2 (55%), which was similar to T1 (59%). However, there wasno difference (P < 0.05) between T1 (59%) and T2 (55%) treatments. Primordial follicles maintained greatermorphological integrity when compared to developing follicles, regardless of the treatment. Based on theseresults, it can be concluded that it is possible to maintain the morphological integrity of PAF in ovaries of catstransported at 4°C for 24 h using TCM 199 and that primordial follicles have a higher viability compared todeveloping follicles.

Assuntos

RESUMO

The objective of this study was to determine the effect of different centrifugation forces in bovine sperm separation by discontinuous Percoll gradients for in vitro fertilization IVF. The semen samples from each bull were pooled or each bull were centrifuged separately and centrifuged in discontinuous Percoll gradients (30, 60 and 90%) at different forces: F1 (9000×g), F2 (6500×g), F3 (4500×g) and F4 (2200×g), according experiment. The sperm samples were evaluated to determine the concentration, motility, vigor, morphology, reactive oxygen species (ROS), integrity of the plasma membrane, lipid peroxidation, antioxidants and embryo development were also evaluated. No difference was observed in the concentration of sperm submitted to different centrifugation forces. The total percentage of motile sperm was increased after centrifugation at F3 and F4, and the ROS production at F1 was greater than the other forces. When the bulls semen were processed individually, no significant differences were observed for the sperm quality parameters between F1 and F4, including lipid peroxidation, antioxidants, cleavage rate and average time to the first cleavage. This work demonstrated for the first time that centrifugation at 2200×g enhanced the sperm penetration and fertilization rates without reducing sperm recovery compared to the typical centrifugation force (9000×g) currently used by the commercial bovine IVF industry.

Assuntos