3种对虾精子超低温冷冻保存技术研究

Cryopreservation of spermatozoa of three prawn species

作者以日本囊对虾(Marsupenaeus japonicus)、中国明对虾(Fenneropenaeus chinensis)和南美白对虾(Penaeus vannamei)作为实验材料,研究3种稀释液(天然海水,人工无钙海水,Hank’s平衡盐溶液)、3种抗冻剂(二甲基亚砜、丙二醇和丙三醇)、3种稀释比例(5%、10%和15%)、3种不同的降温程序对3种对虾精子超低温冷冻保存效果的影响。冻精解冻后通过台盼蓝染色的方法计算其存活率。结果显示,对于3种对虾,以天然海水作为稀释液,10%PG作为抗冻剂,4℃平衡20 min,以–2℃/min降温至–40℃,–40℃平衡10℃,以–15℃/min降温至–150℃,然后投入液氮作为降温程序进行超低温冷冻保存效果最好,冻精解冻后存活率分别为75.33%±2.56%、69.00%±3.00%和23.33%±8.55%。

This study was conducted to examine the cryoprotective effects of three extenders(natural seawater,calcium-free saline,and Hank’s balanced salt solution),three cryoprotectants(DMSO,propylene glycol,and glycerol)at different concentrations and three dilution ratios(5%,10%,and 15%),and three cooling procedures on the cryopreservation of sperms of three types of shrimp,Kuruma prawn,Penaeus japonicus,and Litopenaeus vannamei.The survival rates of post-thawed sperms were calculated by trypan blue staining,and the cryopreservation procedures of the three shrimps were screened.Successful cryopreservation of sperms of these three shrimps in liquid nitrogen was achieved using sterilized natural seawater containing 10%propylene glycol with A-1 procedure,with the survival rates being 75.33%±2.56%,69.00%±3.00%,and 23.33%±8.55%,respectively.