[目的]试验旨在研究稀释液中添加黄芪多糖对杜泊种公羊精液冷冻保存效果的影响.[方法]使用假阴道法采集6只成年杜泊种公羊的精液,将活率＞80％的精液混合,使用Tris-果糖-海藻糖-柠檬酸基础稀释液和降温稀释液对绵羊新鲜精液进行逐步稀释,低温缓慢降温到4 ℃后,分别于含黄芪多糖(0、0.01、0.05和0.1 mg/mL)的冷冻稀释液中进行平衡处理,将平衡后的精液分装于0.25 mL冻精细管中,置于液氮上方熏蒸后于液氮中冷冻保存.通过精子辅助分析系统检测冷冻-解冻后精子质量指标(活力和活率)和精子运动参数(平均路径速度、曲线运动速度、直线运动速度、摆动指数、线性度和直线运动指数),采用精子低渗肿胀试验检测质膜完整性,使用双荧光染色检测精子DNA完整性和线粒体活性.[结果]精液冷冻前后,绵羊精子活力、活率和运动能力均明显降低.0.01 mg/mL黄芪多糖添加组精液冷冻解冻后精子活力、活率、平均路径速度、线性度、摆动指数以及线粒体活性均显著高于其余各组(P＜0.05);0.01 mg/mL黄芪多糖组精子曲线运动速度、直线运动速度、直线运动指数以及质膜完整性和DNA完整性均显著高于对照组和0.10 mg/mL黄芪多糖组(P＜0.05),与0.05 mg/mL黄芪多糖组间差异不显著(P＞0.05).[结论]冷冻稀释液中添加0.01 mg/mL黄芪多糖能够保护绵羊精子结构,有效提高冷冻-解冻后绵羊精子活力、活率和运动能力,改善绵羊精液冷冻保存效果.
[Objective]The experiment was aimed to investigate the effect of adding Astragalus polysaccharide into the diluent on the cryopreservation of Dorper ram semen.[Method]The semen was collected from six adult Dorper rams by artificial vagina method,and semen with a viability rate＞80％was mixed.The fresh semen of sheep was gradually diluted using the base diluent and the cooling diluent,and it was slowly cooled down to 4 ℃,then the semen was diluted with the added Astragali polysaccharides of 0,0.01,0.05,and 0.10 mg/mL and cryo-equilibrated.The balanced semen was dispensed into 0.25 mL freezing tubule and fumigated above liquid nitrogen before semen were frozen at liquid nitrogen.The sperm quality indicators(vitality and viability)and sperm motility parameters(velocity average path,velocity curvilinear,velocity straight line,wobble,linearity,and straightness)were detected through CASA after freezing-thawing.The plasma membrane integrity was detected by HOST,and the sperm DNA integrity and mitochondrial activity were detected using dual fluorescence staining.[Result]Before and after semen freezing,the sperm motility,viability and motility of sheep were significantly reduced.Sperm vitality,viability,velocity average path,linearity,wobble and mitochondrial activity were significantly higher in 0.01 mg/mL Astragali polysaccharides-added group than in the rest of the groups after semen freezing and thawing(P＜0.05).The sperm velocity curvilinear,velocity straight line,straightness,plasma membrane integrity,and DNA integrity of 0.01 mg/mL Astragalus polysaccharide group were significantly higher than those of control group and 0.10 mg/mL Astragalus polysaccharide group(P＜0.05),but had no significant difference with 0.05 mg/mL Astragalus polysaccharide group(P＞0.05).[Conclusion]The addition of 0.01 mg/mL Astragalus polysaccharide to the freezing diluents was able to protect the structure of sheep spermatozoa,effectively increase sperm viability,viability and motility after freezing-thawing,and improve the effect of sheep semen freezing.
王振国;李瀚森;张丽;周千;时娟娟;石磊;任有蛇
山西农业大学动物科学学院,太谷 030801
畜牧业
杜泊羊;精液;冷冻保存;黄芪多糖
Dorper sheep;semen;cryopreservation;Astragalus polysaccharides
《中国畜牧兽医》 2024 (005)
2007-2014 / 8
山西省应用基础研究计划资助项目(201901D111234、20210302123363);山西省现代农业羊产业技术体系资助项目(2023CYJSTX14)
10.16431/j.cnki.1671-7236.2024.05.022
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