大鳞副泥鳅精液超低温冷冻保存方法的优化

乔瑞峰 1; 陆专灵 2; 高爽爽 1; 雷　坤 1; 韦友传 1

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乔瑞峰 1，陆专灵 2，高爽爽 1，雷　坤 1，韦友传 1.大鳞副泥鳅精液超低温冷冻保存方法的优化[J].广东农业科学,2020,47(7):142-147

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Optimization of Method for Ultra-low Temperature Cryopreservation of Paramisgurnus dabryanus Semen

DOI：10.16768/j.issn.1004-874X.2020.07.018

中文关键词: 大鳞副泥鳅精液保存超低温冷冻精子活力

英文关键词: Paramisgurnus dabryanus semen preservation ultra-low temperature freezing sperm motility

基金项目:广西创新驱动发展专项（桂科 AA17204094-8）

作者	单位
乔瑞峰 1，陆专灵 2，高爽爽 1，雷　坤 1，韦友传 1	1. 广西大学动物科学技术学院，广西南宁 530004；2. 广西水产科学研究院 / 广西水产遗传育种与健康养殖重点实验室，广西南宁 530021

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中文摘要:

【目的】优化大鳞副泥鳅精液超低温冷冻保存方法。【方法】以解冻后的精子活力为参数，调控试验条件，筛选最佳精液稀释液和最佳抗冻剂，优化抗冻剂体积分数和精液稀释比例。【结果】5 种稀释液中，稀释液Ⅳ效果最好，冻精活力为 35.5（±2.7）%；甘油（GLY）、甲醇（METH）、二甲基亚砜（DMSO）等 3种抗冻剂中，5%（V/V）DMSO 处理的冻精活力为 40.7（±2.5）%，显著高于 5%GLY 和 METH 处理的冻精活力；优化 DMSO 的体积分数发现，10% 终浓度的 DMSO 对精液保护效果较好，冻精活力为 54.6（±1.5）%；优化稀释液Ⅳ与精液稀释比例发现，稀释液Ⅳ与精液以 3 ∶ 1 体积比稀释时能达到良好效果，冻精活力可达 65.4（±2.3）%。【结论】精液稀释液Ⅳ与精液体积比为 3 ∶ 1，终浓度 10% DMSO 为抗冻剂，对大鳞副泥鳅精液超低温冷冻保存的效果最佳。

英文摘要:

【Objective】The study was to optimize the method for ultra-low temperature cryopreservation of the semen of Paramisgurnus dabryanus.【Method】With the thawed sperm motility as the parameter, the experimental conditions were controlled, the best semen diluent and antifreeze were selected, and the volume fraction of antifreeze and the dilution ratio of semen were optimized.【Result】Among the five diluents, diluent Ⅳ was the most effective, with a frozen sperm motility of 35.5（±2.7）%. For the three antifreeze protectants, namely, glycerol（GLY）, methanol（METH）, and dimethyl sulfoxide（DMSO）, when treated with 5%（V/V）DMSO, the frozen sperm motility was 40.7（±2.5）%, significantly higher than that of the 5% GLY and METH treatments. Optimization of the DMSO volume fraction revealed that 10% final concentration of DMSO was more effective in protecting semen, with a frozen sperm motility of 54.6（±1.5）%. Based on the optimization of dilution Ⅳ and semen dilution, it was found that good results could be achieved when diluent IV was diluted with semen at a volume ratio of 3 ∶ 1, and the frozen sperm motility could reach up to 65.4（±2.3）%.【Conclusion】By setting the volume ratio of diluent Ⅳ to the semen as 3 ∶ 1 and a final concentration of 10% DMSO as antifreeze, the best results were obtained when the semen of P. dabryanus was cryopreserved at ultra low temperature.

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