一种超低温冷冻蜜蜂精液方法的研究

Study on a Method of Cryopreserving Bee Semen

为了减少冷冻保护剂对蜜蜂精液的毒害作用提高冷冻保存后蜜蜂精子活率和活力,试验组采用精液与稀释剂先4℃静置1h再与冷冻保护剂4℃静置1 h的平衡方法,用程序降温仪从4℃以3.5℃/min下降速率下降到-60℃的冷冻方法;对照组采用稀释剂、冷冻保护剂与精液同时混合4℃静置2h的平衡方法和在液氮上方2~3cm处快速降温的冷冻方法,比较两组保存1d、7d、60d、180 d和360 d的精子活率、活力和授精蜂王存活率。结果表明保存60 d、180 d和360 d的精子活率、活力和授精蜂王存活率,试验组均显著高于对照组(P<0.05)。说明试验组的冷冻蜜蜂精液方法可同时满足蜜蜂精子长期冻存和短期冻存的需求,可以作为解决蜜蜂优良蜂种遗传物质传递的一种方法。

In order to reduce the toxic effect of cryoprotectant on bee semen and improve the viability and vitality of bee sperm after cryopreserved,two methods of cryopreserved bee semen were used.The experiments were divided to the experimental group and control group,the semen and the diluent were placed at 4℃for 1 h,and then frozen with a cryoprotectant at 4℃for 1 h.After that,the cryoprotectant was used to refrigerate the semen at a rate of 3.5℃/min from 4℃to-60℃.In the control group,the balance method of diluent,cryoprotectant and semen were mixed at 4℃for 2 h,and the freezing method of rapid cooling at 2~3 cm above liquid nitrogen was adopted.The sperm viability,vitality and the survival rate of the inseminated queen bee were compared between the two groups after 1 day,7 days,60 days,180 days and 360 days.The results showed that the sperm viability,viability and survival rate of queen bee after 60 days,180 days and 360 days in the experimental group were significantly higher than that in the control group(P<0.05).This method can meet the needs of both longterm and short-term cryopreserving of honeybee sperm,and can be used as a method to solve the genetic material transfer of superior honeybee species.