在4℃降温平衡过程中0.2g/L咖啡因与精子共孵育时间对猪颗粒冻精质量的影响

Effects of 0.2g/L caffeine and sperm co-incubation time on the quality of boar semen frozen pellets during the cooling process at 4℃

实验旨在探讨在猪精液于4℃平衡2 h过程中0.2 g/L咖啡因与精液共孵育2 h、1 h和0.5 h,对颗粒冻精解冻后精子活率、活力、质膜完整性和顶体完整性以及解冻后精子体外存活时间等指标的影响,以期进一步提高猪颗粒冻精质量。实验结果表明,在精液冷冻之前,咖啡因不同平衡时间组的精子活率和活力都呈现出升高趋势,但与对照组差异不显著;咖啡因与精液共孵育2 h、1 h和0.5 h组的精子顶体完整率和质膜完整率显著低于对照组,前3组之间差异均不显著。而颗粒冻精解冻后,咖啡因与精液共孵育2 h、1 h和0.5 h组精子活率和活力均显著高于对照组,其中共孵育1 h组显著高于其他3组(P<0.05),共孵育2 h组和0.5 h组间差异不显著(P>0.05);共孵育2 h组精子顶体完整率和质膜完整率显著低于对照组和共孵育0.5 h、1 h组(P<0.05),但后3组之间差异不显著;共孵育2 h组精子存活时间显著低于对照组、共孵育1 h和0.5 h组(P<0.05),其中共孵育1 h组精子存活时间最长,达7 d以上。总之,在精液4℃降温平衡过程中咖啡因与精液共孵育1 h对冷冻后精子质量最有利,解冻后精子活率和活力显著高于其他各组,且解冻后精子体外存活时间最长。

In order to further improve the quality of boar frozen semen pellet,the present study discussed the effect of the co-incubation time of 0.2 g/L caffeine and semen for 2 h,1 hand 0.5 h on frozen thawed sperm viability,motility,plasma membrane integrity,acrosome integrity and survival time during the cooling process at 4℃.The sperm viability and motility in the different groups caffeine incubation exhibited an increasing trend as showed in our experiment,but there was no significant difference when comparing with the Control.The acrosome integrity rate and plasma membrane integrity rate were significantly higher in the control group than those in the groups of 2 h,1 h and 0.5 h caffeine incubation,however,the differences among the latter three groups showed no significance.After the semen were frozen,the sperm viability and motility in the caffeine incubation groups were significantly higher than those in the control group,among which the group of 1 h incubation was significantly higher than the other three groups(P<0.05),and there were no significant difference between 2 h incubation group and 0.5 h incubation group(P>0.05).As for acrosome integrity rate and plasma membrane integrity rate,the group of 2 h incubation was significantly lower than that in the control group and the groups of 0.5 h and 1 h incubation(P<0.05),nevertheless,there showed no significant difference of sperm acrosome integrity rate and plasma membrane integrity rate among the latter three groups.The sperm survival time in the group of 2 h incubation was significantly lower than that in the control,the 1 h and 0.5 h incubation groups(P<0.05),in which the survival time in the 1 h incubation group was the longest and more than 7 d.In conclusion,the 1 h incubation of caffeine and sperm was the most beneficial to sperm quality after frozen during the cooling process at 4℃,and sperm viability and motility of the frozen sperm were significantly higher than those in other groups,and besides,the sperm survival time of frozen sperm in the group was the longest in vitro.