不同冷冻保护液和解冻温度对猪精液冷冻效果的影响

Eeffects of different cryoprotectants and thawing temperatures on cryopreservation of boar spermatozoa

在BF5稀释液中分别添加不同水平的牛血清白蛋白(BSA)(0.5,1,1.5g/L)、二甲基乙酰胺(DMA)(0.5%,1%,1.5%)、甲基-β-环糊精载胆固醇(CLC)(1,2.5,5g/L),对成年健康猪精液进行冷冻保存,于不同温度(37,50,70℃)解冻后分别检测冷冻后精子的活率、活力、质膜完整性、顶体完整率、线粒体活性。结果显示,0.5g/LBSA组冷冻后精子活力、质膜完整率、顶体完整率和线粒体活性均高于其他组,但差异不显著(P>0.05)。1%DMA组冷冻后精子活率和活力显著优于1.5%DMA组(P<0.05),同时也高于对照组(3%甘油)冻后精子活率和活力,但差异不显著。不同质量浓度CLC组冷冻后精子的活率、活力、质膜完整性和线粒体活性与对照组相比差异不显著。50℃和70℃解冻组精子的活率和线粒体活性显著高于37℃解冻组的精子;50℃解冻组精子的活力和质膜完整率显著高于其他2组。因此,在猪精液冷冻稀释液中,用DMA可以代替甘油作为渗透性保护剂,且以1%DMA、50℃解冻最佳。

To explore the effects of several different concentrations of additives on cryopreservation of boar spermatozoa,four experiments were designed for the detection of different concentrations （0.5,1,1.5 g/L） of bovine serum albumin （BSA）,dimethyl acetamide （DMA） （0.5%,1%,1.5%）,cholesterol-loaded methyl-β-cyclodextrin （CLC） （1,2.5,5 g/L） and thawing temperatures （37,50,70℃）.The sperm viability,motility,membrane integrity,acrosome integrity and mitochondrial activity were examined post-thawing.The results showed that sperm viability,membrane integrity,acrosomal integrity and mitochondrial activity for 0.5 g/L BSA were higher than other group,but no significant difference was observed （P 0.05）.Sperm viability and motility for 1% DMA was superior （P0.05） to 1.5% DMA,with no significant difference between 1% DMA and 3% glycerol.Sperm viability,motility,membrane integrity and mitochondrial activity for all different concentrations of CLC were not significantly different compared to the control group.When sperm were thawed at 50℃and 70℃,the sperm viability and mitochondrial activity was superior （P0.05） to those at 37℃.Sperm motility and membrane integrity for 50℃ thawing were higher （P0.05） as compared to the others.In conclusion,boar semen was successfully cryopreserved by replacement of glycerol with DMA （especially 1% DMA） and thawing at 50℃ was better.