咖啡因对猪精液冷冻的影响及冷冻-解冻方法的优化  

Effects of Caffeine on the Cryopreservation of Boar Semen and Optimizationg of Freezing and Thawing Protocal of Rapid Freezing Techniques

作　　者：张渭斌[1] 刘丑生[2] 孙世铎[1] 于福清[2] 赵俊金[2] 孟飞[2] 史建民[2] 谷合勇[3] 

机构地区：[1]西北农林科技大学动物科技学院,杨凌712100 [2]农业部种畜品质监督检测中心,北京100193 [3]云南农业大学动物科技学院,昆明650201

出　　处：《中国畜牧兽医》2010年第10期148-152,共5页China Animal Husbandry & Veterinary Medicine

基　　金：农业部"十一五"科技支撑计划课题:荣昌猪资源保护体系及关键技术研究(2007BAD51B01);优良种猪的高效利用及胚胎冷冻保存技术研究

摘　　要：本试验对猪精液冷冻保护液中添加咖啡因的作用进行了研究,并优化冷冻-解冻程序,提高0.5 mL细管猪精液冷冻解冻后的质量。在预先设计稀释液配方的基础上,使咖啡因终浓度为0、0.2、0.4、0.6 mg/mL,选择最佳咖啡因浓度,选出最优组合与传统的TCG稀释液和商业用Androhep CryoGuardTM冷冻稀释液进行比较,比较3种不同的冷冻曲线对猪冷冻精液解冻后精子品质的影响,最后对38℃下30 s、50℃下13 s 2种解冻方法进行比较。结果表明,咖啡因浓度为0.2、0.4mg/mL的精子冷冻后活力、质膜完整率、顶体完整率显著高于00、.6 mg/mL(P<0.05),最佳添加浓度为0.2 mg/mL;预设计的稀释液配方及冷冻方案优于传统的TCG法(P<0.05),但与Androhep CryoGuardTM稀释液(美国商业用)有一定差距(P<0.05);A曲线在猪精液冷冻-解冻后活力、质膜完整率和顶体完整率方面均显著优于B和C冷冻曲线(P<0.05);50℃水浴解冻13 s显著优于38℃解冻30 s(P<0.05)。The aim of this study was to investigate the effect of supplementation of caffeine in freezing extender on frozen-thawed semen quality.Optimize freezing and thawing protocol in 0.5 mL straws.Supplementation of caffeine directly into preliminary design freezing extender,the concentration of caffeine was 0,0.2,0.4,0.6 mg/mL,determined the best concentration of caffeine.Then compared with the traditional TCG extender and the AndrohepCryoGuardTM extender,comparison of three different freezing curves on the frozen-thawed sperm quality.The frozen sperm samples were thawed by warming them at 37 ℃ water bath for 30 s and 50 ℃ water bath for 13 s.The results showed that there was a significantly higher percentage of progressive motility,membrane integrity and acrosome integrity in caffeine supplemented groups than control group（P〈0.05）,the best concentration was 0.2 mg/mL.The preliminary design was better than the TCG（P〈0.05）,but it had a significant variance with AndrohepCryoGuardTM extender（P〈0.05）.Three different freezing curve had different influence on sperm quality parameters.Freezing curve A had a higher percentages of TMS,PMI and NAR of spermatozoa than B and C（P〈0.05）,thawed at 50 ℃ for 13 s was better than 38 ℃ for 30 s.

关 键 词：猪精液 咖啡因 冷冻曲线 冷冻速率 

分 类 号：S814.4[农业科学—畜牧学]