Effect of storing time at 5 degree centigrade prior to freezing on the motility and fertility of bull spermatozoa
1998
Sakai, M. (Iwate Univ., Morioka (Japan). Faculty of Agriculture) | Tauchi, S. | Geshi, M. | Kikuchi, H. | Hashizume, T. | Masuda, H.
The purpose of this experiment was to clarify the effect of strong at 5 degree C prior to freezing on the motility and fertility of frozen-thawed bull spermatozoa. Ejeculated bull semen was diluted immediately 4-5 fold with the 1st diluent at 30 degree C. Diluted semen devided into 5 test tubes were cooled to 5 degree C taking for 2 hr and then stored for 0, 2, 6, 10 and 20 hr until the semen were diluted by the 2nd diluent. The 2nd diluent was added to each stored semen in a single addition just before freezing. The sperm concentration was adjusted to 5 x 10(7) sperm/ml. Each treated semen was filled in 0.5 ml straws and frozen in LN vapour. Thawed semen was incubated for 3 hr at 38 degree C. The motility of spermatozoa just after thawing and after 3 hr incubation at 38 degree C were analyzed by Hamilton-Thorne Research Motility Analyzer (Ver. 10.6). The viability and membrane integrity of frozen-thawed spermatozoa were evaluated by Triple-Stain technique. The fertilizing ability of frozen-thawed spermatozoa was evaluated by using in vitro fertilization technique. 1) The storing time at 5 degree C before freezing required to get the best motility was more than 2 hr. 2) After 3 hr incubation at 38 degree C, the viability of 6, 10 and 20 hr storage was significantly higher than that of 0 and 2 hr, and the membrane integrity of 6 hr storage was significantly higher than that of 0 hrin a bull (Bull-C) (p0.05). 3) The sperm penetration rate of 6 hr storage was significantly higher than that of 0 and 2 hr in a bull (Bull-C) (p0.05). The present study suggested that 6 hr exposure to 5 degree C without glycerol prior to freezing was important for getting the high motility, viability, membrane integrity and fertilizing ability of frozen-thawed bull spermatozoa
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