Efeito de diferentes diluentes na qualidade e fertilidade do sêmen refrigerado de jumento Pêga (Equus asinus)
Carregando...
Data
2020-07-30
Autores
Gobato, Mariana Luiza Mezzena
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
Frente aos resultados menos expressivos no uso de sêmen refrigerado e o interesse no comércio de muares, o aprimoramento das biotecnologias é um passo importante para a espécie asinina. Dentre os principais fatores relacionados com o sucesso no uso do sêmen refrigerado destaca-se a presença do plasma seminal e a composição do diluente utilizado para garantir a viabilidade espermática durante o processo de fertilização. Os objetivos do presente estudo visam avaliar o efeito de três diluentes sobre a qualidade do sêmen asinino, com ou sem plasma seminal, bem como os efeitos da fertilidade in vivo. Foram utilizados 2 ejaculados de 6 jumentos da raça Pêga, entre 4 e 12 anos. Cada ejaculado foi diluído em meios comerciais a base de leite desnatado (BotuSêmen Special®), caseína (BotuSêmen Gold®) e gema de ovo (BotuCrio®) e as amostras processadas na presença e remoção de plasma seminal, conforme os 6 tratamentos: leite, caseína e gema de ovo com plasma na concentração de 50 milhões de espermatozoides/mL; leite, caseína e gema de ovo centrifugado e ressuspendido na concentração de 100 milhões de espermatozoides/mL. As amostras foram avaliadas a fresco (T0), 24 (T24) e 48 (T48) horas pós-refrigeração. Os parâmetros avaliados foram: cinética espermática pelo método computadorizado CASA e desestabilização da membrana, produção de espécies reativas ao oxigênio e atividade mitocondrial por citometria de fluxo. Para o teste de fertilidade foram utilizados 6 ciclos de 15 éguas em sistema crossover e um jumento Pêga com fertilidade comprovada. O diluente à base de leite apresentou resultados inferiores para motilidade total (MT), progressiva (MP) e rápidos (RAP) sem plasma seminal em todos os momentos avaliados e maior produção de ânion superóxido em T48, na presença do plasma seminal (P<0,05). Já o meio à base de caseína sem plasma seminal, apresentou resultados superiores para todos os parâmetros avaliados em todos os momentos (P<0.05), com exceção dos valores de desestabilização de membrana em T24 que não apresentou diferença significativa. Os resultados com meio à base de gema de ovo não sofreram efeito do plasma seminal e também mostraram-se superiores, com exceção de MP e RAP em T48 (P<0,05). A caseína e a gema de ovo apresentaram elevada taxa de fertilidade tanto com (73% vs 93%) quanto sem plasma seminal (93% vs 73%), respectivamente. Já o leite desnatado apresentou valores similares sem plasma seminal (60%), porém diferiu estatisticamente na presença de plasma (20%). Conclui-se que apesar de demonstrar resultados satisfatórios na refrigeração do sêmen equino, o leite desnatado apresenta pior interação com o sêmen asinino refrigerado. Já a gema de ovo e a caseína, demonstraram ser as melhores alternativas para a refrigeração de sêmen asinino por até 48 horas.
In view of the less expressive results using cooled semen and the interest in breeding mules, the improvement of biotechnologies is an important step for the donkey species. Among the main factors related to the successful use of cooled semen, the presence of seminal plasma and the composition of the extender used to guarantee sperm viability during the fertilization process stands out. The aims of this study were to evaluate the effect of three extenders on the quality of donkey semen, with or without seminal plasma, as well as the effects of in vivo fertility. Two ejaculates from 6 donkeys of the Pêga breed (4 -12y), were used. Each ejaculate was extended in commercial extender based on skimmed milk (BotuSemen Special®), casein (BotuSemen Gold®) and egg yolk (BotuCrio®) and the samples processed in the presence and removal of seminal plasma, according to the 6 treatments: milk , casein and egg yolk with plasma at a final concentration of 50 million sperm per mL; milk, casein and egg yolk centrifuged and resuspended at a final concentration of 100 million sperm per mL. The samples were evaluated fresh (T0), 24 (T24) and 48 (T48) hours after refrigeration. The parameters evaluated were: sperm kinetics by the CASA and membrane destabilization, reactive oxygen species production and mitochondrial activity by flow cytometry. For fertility test, 6 cycles of 15 mares were used in a crossover system and semen from a Pêga donkey with proven fertility. Milk-based extender had lower results for total (TM), progressive (PM) and rapid (RAP) motility without seminal plasma at all evaluated times and higher production of superoxide anion in T48, in the presence of seminal plasma (P < 0.05). Casein-based extender without seminal plasma had superior results for all parameters evaluated at all times (P <0.05), except for the values of membrane destabilization at T24, with no statistical difference. Results of egg yolk-based extender were not affected by seminal plasma and were also superior, with the exception of PM and RAP at T48 (P<0.05). Casein and egg yolk had a high fertility rate with (73% vs 93%) and without seminal plasma (93% vs 73%), respectively. On the other hand, skimmed milk had similar values without seminal plasma (60%), but it differed statistically in the presence of plasma (20%). In conclusion, despite satisfactory results for equine cooled semen, skimmed milk extender had a worse interaction with donkey cooled semen. Egg yolk and casein proved to be the best alternatives for cooling donkey semen for up to 48 hours.
In view of the less expressive results using cooled semen and the interest in breeding mules, the improvement of biotechnologies is an important step for the donkey species. Among the main factors related to the successful use of cooled semen, the presence of seminal plasma and the composition of the extender used to guarantee sperm viability during the fertilization process stands out. The aims of this study were to evaluate the effect of three extenders on the quality of donkey semen, with or without seminal plasma, as well as the effects of in vivo fertility. Two ejaculates from 6 donkeys of the Pêga breed (4 -12y), were used. Each ejaculate was extended in commercial extender based on skimmed milk (BotuSemen Special®), casein (BotuSemen Gold®) and egg yolk (BotuCrio®) and the samples processed in the presence and removal of seminal plasma, according to the 6 treatments: milk , casein and egg yolk with plasma at a final concentration of 50 million sperm per mL; milk, casein and egg yolk centrifuged and resuspended at a final concentration of 100 million sperm per mL. The samples were evaluated fresh (T0), 24 (T24) and 48 (T48) hours after refrigeration. The parameters evaluated were: sperm kinetics by the CASA and membrane destabilization, reactive oxygen species production and mitochondrial activity by flow cytometry. For fertility test, 6 cycles of 15 mares were used in a crossover system and semen from a Pêga donkey with proven fertility. Milk-based extender had lower results for total (TM), progressive (PM) and rapid (RAP) motility without seminal plasma at all evaluated times and higher production of superoxide anion in T48, in the presence of seminal plasma (P < 0.05). Casein-based extender without seminal plasma had superior results for all parameters evaluated at all times (P <0.05), except for the values of membrane destabilization at T24, with no statistical difference. Results of egg yolk-based extender were not affected by seminal plasma and were also superior, with the exception of PM and RAP at T48 (P<0.05). Casein and egg yolk had a high fertility rate with (73% vs 93%) and without seminal plasma (93% vs 73%), respectively. On the other hand, skimmed milk had similar values without seminal plasma (60%), but it differed statistically in the presence of plasma (20%). In conclusion, despite satisfactory results for equine cooled semen, skimmed milk extender had a worse interaction with donkey cooled semen. Egg yolk and casein proved to be the best alternatives for cooling donkey semen for up to 48 hours.
Descrição
Palavras-chave
Asinino, Refrigeração, Caseína, Gema de ovo, Leite desnatado, Asinine, Cooled semen, Casein, Egg yolk, Skimmed milk