猪精液冷冻技术的研究

Studies on Boar Semen Freezing Techniques

以解冻后的精子活率、活力、质膜完整率、顶体完整率和人工授精结果为判定指标 ,比较了几种冷冻稀释液、解冻液及冷冻 -解冻程序对猪精液冷冻的效果 ,并对所筛选出的最佳稀释液和冷冻程序做了改进。结果如下 :(1) 号稀释液冷冻解冻后的精子活力 (32 .4± 7.3) %、活率 (4 2 .2± 3.2 ) %、顶体完整率 (6 2 .3± 0 .8) %和弯尾率 (4 2 .6± 7.5 ) %均显著高于 、 、 号稀释液 (P<0 .0 5 )。 (2 )在 号稀释液中添加 0 .5 %、1%和 2 %的 O.E.P(氨基 -钠 -十二烷硫酸酯 )使精子活率提高 5 %、弯尾率提高 6 %、顶体完整率提高近 10 % ,与对照组相比差异显著 (P<0 .0 5 )。 (3)应用改进的稀释液 ( 液加 1% O.E.P) ,细管法比颗粒法冷冻精子活率提高近 7个百分点 ,活力提高 6个百分点 (P<0 .0 5 )。(4 )室温预平衡 4h的精子活力和活率都比对照组 (0 h)有显著提高 (P<0 .0 5 ) ,而预平衡 2、6 h的精子活力和活率与对照组差异不显著 (P>0 .0 5 )。 (5 ) 号解冻液解冻后精子活力 (4 2 .9± 2 .6 ) %显著高于 、 、 号解冻液 (P<0 .0 5 )。 (6 )用 5 0℃水浴解冻精子的活力 (4 6 .3± 2 .6 ) %显著高于 39℃ (4 2 .9± 2 .6 ) %和 70℃水浴 (4 1.1± 5 .7) %解冻的精子活力 (P<0 .0 5 )。 (7)精清和 号解冻液?

Effects of several extenders,thawing diluents and freezing-thawing protocols on boar sperm viability,motility,plasma membrane integrity and acrosome integrity were studied in this experiment and the best extender and protecol were modified to improve boar semen freezing efficiency.The results were as follows:(1)The number Ⅳ extender was the best among the extenders tested and its sperm motility (32.4±7.3)%,vaibility (42.2±3.2)%,acrosomal integrity (62.3±0.8)% and plasma membrane integrity (42.6±7.5)% after freezing and thawing were significantly(P<0.05) higher than those obtained with number Ⅰ,Ⅱ and Ⅲ extenders. (2)When 0.5%,1% or 2% O.E.P. were added to the number Ⅳ extender,sperm viability,plasma membrane integrity and acrosomal integrity increased significantly(P<0.05) by 5%,6% and 10%,respectively,over those of the controls. (3)When boar semen was frozen in straws with modified extender(number Ⅳ plus O.E.P.),the sperm motility and viability were increased by 7% and 6% over those when semen was frozen in pellets with the same extender(P<0.05). (4)Pre-equiliberation for 4 hours at room temperature increased both sperm viability and motility significantly(P<0.05) in comparison with those in the controls(0 hours of pre-equilibrium). (5)The sperm motility obtained with thawing diluent number Ⅲ(42.9±2.6)% was significantly higher than those obtained with thawing diluents number Ⅰ,Ⅱ and Ⅳ. (6)The sperm motility after 50℃ water bath thawing(46.3%) was significantly higher than those obtained with 39℃(42.9±2.6)% and 70℃ water bath thawing(41.1±5.7)%. (7)When the frozen semen was thawed in semen plasma,the sperm motility,viability,plasma membrane integrity and acrosomal integrity was not significantly different from those when semen was thawed in thawing diluent number Ⅲ. (8)All of the 6 sows inseminated with the frozen-thawed semen with a motility of around 46%(which was processed with modified extender,pre-equiliberated for 4 hours and thawed in thawing diluent number Ⅲ at 50℃) got pregnant and farrowed at an average liter size of 9.