Effect of sperm separation techniques on the viability and acrosome reaction of frozen-thawed ovine spermatozoids

[摘要] The effectiveness of four sperm separation techniques on thawed ovine semen was evaluated. A hundred and sixty straws with 0.5 ml of semen with a concentration of 200x106/ml were used. In each work session, four straws were thawed at 37.5ºC for 45 sec mixing their contents. Then, 200 µl were taken to perform one of the following techniques: Swim-up, Percoll, filtering through glass fiber or filtering through Sephadex. Increased progressive motility of spermatozoids was obtained through swim-up, glass fiber and Sephadex (56.1%, 66.4%, and 56.8%, respectively), which was significantly different (p<0.05) in those recovered from glass fiber. The numbers of live spematozoids with acrosome reaction were high with all the separation techniques, with a significant difference (p<0.05) in those obtained from swim-up, glass fiber and Sephadex, whose values were higher than 50.0%. The numbers of dead spermatozoids, with and without acrosome reaction, were low in all the techniques, with the exception of those obtained by Percoll, which was significantly different (p<0.05) to the control, excepting those obtained by Sephadex and swim-up when the dead ones with acrosome reaction were evaluated. When evaluating live spermatozoids without acrosome reaction, a low value of this indicator was found in all the treatments (13.1, 9.1, 9.3 and 10.8%, swim-up, Percoll, glass fiber and Sephadex, respectively. The sperm viability evaluated with the hypo-osmotic test and propidium iodide concluded that a higher percentage of live spermatozoids were obtained when they were filtered with glass fiber (73.0% and 68.3%, hypo-osmotic test and propidium iodide, respectively); no significant differences (p<0.05) were observed between these two last test.

[学科分类] 兽医学

[关键词] swim-up;sperm separation techniques;ovine spermatozoids