RESUMO

The ex situ reproduction of wild animals still faces several obstacles, such as minimum reproductive rates or even reproductive inactivity, data that makes specialists worried considering the growing number of species threatened with extinction. Reproductive biotechniques are already well developed among farm animals and widely used in conservation programs, but studies are still needed to develop protocols adapted for snakes, regarding their wide variety of species and their particularities. Therefore, this work seeks to compare sperm survival over time in Oxyrhopus guibei when kept at 24°C and under refrigeration. For this, sperm analysis will be made by evaluating parameters such as motility, progressive motility, presence of morphological changes and mitochondrial activity in spermatozoa. Three males of O. guibei were used, belonging to the Ecology and Evolution Laboratory of the Butantan Institute, and semen was collected via digital stimulation without the use of local anesthesia. This study showed that it is possible to maintain O. guibei sperm cells in HAM-F10 cell culture medium for up to 8 hours at 24°C and for up to 48 hours under refrigeration at 4°C.

A reprodução de animais silvestres ex situ ainda enfrenta diversos obstáculos, como taxas reprodutivas mínimas ou até mesmo inatividade reprodutiva, dados que preocupam especialistas considerando o crescente número de espécies ameaçadas de extinção. As biotécnicas reprodutivas são bem desenvolvidas entre animais de produção e amplamente empregadas em programas de conservação, porém ainda carecem estudos para desenvolvimento de protocolos adaptados para serpentes, tendo em vista a grande variedade de espécies e suas particularidades. Dessa forma, este trabalho busca comparar a sobrevivência espermática ao longo do tempo em Oxyrhopus guibei quando mantidas a 24°C e sob refrigeração. Para isso, foi feita a análise espermática avaliando parâmetros como motilidade, motilidade progressiva, presença de alterações morfológicas e a atividade mitocondrial em células espermáticas. Foram utilizados 3 machos de O. guibei, pertencentes ao plantel do Laboratório de Ecologia e Evolução do Instituto Butantan e foi feita a colheita de sêmen via estimulação digital sem uso de anestesia local. Este estudo mostrou que é possível manter espermatozoides de O. guibei em meio HAM-F10 por até 8 horas a 24°C e por até 48h sob refrigeração, a 4°C.

RESUMO

Comparar la sobrevida y la movilidad de espermatozoides incubados en medios HAM’S-F10 y G-IVF. Estudio prospectivo. Se incubaron submuestras de espermatozoides recuperados por gradientes de densidad de 31 hombres normospérmicos, en los medios respectivos por 18 a 20 horas en una atmósfera de 5 por ciento CO2. Posteriormente se evaluaron la sobrevida y la movilidad espermáticas. Las direfencias encontradas se evaluaron mediante la t de Student para muestras apareadas con transformación arcoseno. Unidad de Fertilidad Unifertes, Clínica El Ávila. Caracas, Venezuela. La sobrevida y la movilidad progresiva espermáticas fueron significativamente mayores en el grupo tratado con G-IVF (P<0,01). Consecuentemente, la media del porcentaje de espermatozoides inmóviles fue significativamente menor (P<0,01). La sobrevida y la movilidad progresiva en espermatozoides incubados con G-IVF fueron significativamente mayores que con HAM’S-F10

To compare survival and motility of sperm incubated in HAM’S-F10 and G-IVF media. Subsamples of motile sperm recovered by density gradients of 31 normospermic men were incubated in each medium for 18-20 hours in 5 percent CO2 atmosphere. Subsequently, sperm survival and motility were assessed. Unidad de Fertilidad Unifertes, Clínica El Ávila. Caracas, Venezuela. Average sperm survival and progressive motility were significantly higher in G-IVF (P<0.01). Consequently, average percentage of immotile sperm was significantly lower (P<0.01). Sperm survival and progressive sperm motility were significantly higher in G-IVF medium than in HAM’S-F10.

Assuntos

RESUMO

Difficulties to be overcome in the widespread use of artificial insemination (AI) in mares are low sperm survival and poor sperm quality, which are encountered frequently among breeding stallions. Therefore, a method is needed to prolong the useable life of stallion spermatozoa destined for AI. In a preliminary study using 8 ejaculates from one stallion, density gradient centrifugation or centrifugation through a single layer of silica colloid appeared to prolong sperm motility compared to uncentrifuged spermatozoa, thereby potentially extending the useable life of treated stallion spermatozoa for AI. Furthermore, there was an improvement in sperm morphology, with the number of morphologically normal spermatozoa increasing from 42 to 60.5% and with the removal of approximately 60% spermatozoa with head or tail defects from the original population. No difference between the two centrifugation methods, in terms of yield or duration of spontaneous motility, could be detected in this study. Either of these methods of colloidal centrifugation could be a useful aid to preparing stallion spermatozoa for artificial breeding techniques, including AI.

Assuntos

RESUMO

Difficulties to be overcome in the widespread use of artificial insemination (AI) in mares are low sperm survival and poor sperm quality, which are encountered frequently among breeding stallions. Therefore, a method is needed to prolong the useable life of stallion spermatozoa destined for AI. In a preliminary study using 8 ejaculates from one stallion, density gradient centrifugation or centrifugation through a single layer of silica colloid appeared to prolong sperm motility compared to uncentrifuged spermatozoa, thereby potentially extending the useable life of treated stallion spermatozoa for AI. Furthermore, there was an improvement in sperm morphology, with the number of morphologically normal spermatozoa increasing from 42 to 60.5% and with the removal of approximately 60% spermatozoa with head or tail defects from the original population. No difference between the two centrifugation methods, in terms of yield or duration of spontaneous motility, could be detected in this study. Either of these methods of colloidal centrifugation could be a useful aid to preparing stallion spermatozoa for artificial breeding techniques, including AI.(AU)

Assuntos