双斑东方鲀精子冷冻保存方法探究

双斑东方鲀(Takifugu bimaculatus)在福建已形成较大规模的养殖产业,但其精卵成熟同步率低、自然交配受精率低等繁殖特点已成为养殖规模化发展的最大制约因素。而精子超低温冷冻保存技术作为直接保存细胞遗传物质的一种有效方法,为双斑东方鲀规模化人工繁育问题的解决提供了方案。研究以复苏后的精子活力为选择指标,开展双斑东方鲀精子冷冻保存方法的研究,对抗冻剂、基础液、精子稀释浓度和降温程序等几个关键影响因素进行了筛选,获得了双斑东方鲀精子的最适冷冻方法:以含5%二甲亚砜(DMSO)的MPRS溶液与精子按1∶1比例稀释,样品在4℃冰箱中平衡30 min,液氮上方10 cm处熏蒸5 min,再下降到5 cm处熏蒸5 min,之后投入液氮。按此方法保存的双斑东方鲀精子在解冻激活后,其活力可达(83±3)%,与相应卵子受精,可得到平均70%的孵化率,可以满足双斑东方鲀规模化人工繁育的需求。研究结果也可为其他东方鲀鱼类的相关研究提供参考。

Research on cryopreservation of Takifugu bimaculatus spermatozoa

Takifugu bimaculatus aquaculture industry has been largely developed in Fujian Province,however, its benign development was restricted by some of its reproductive characters like the unsynchronization of maturity of sperm and egg,and low fertility rate under nature mating conditions.Sperm cryopreservation method provides an effective solution for this restriction because it can preserve germplasm directly. In this study,the optimal parameters for sperm cryopreservation of T.bimaculatus were tested.The most suitable treatment for T. bimaculatus sperm is to dilute the sperm by MPRS solution with 5 % DMSO at a ratio of 1 : 1,freeze at 4 ℃for 30 min,fumigate at 10 cm above the surface of liquid nitrogen for 5 min and then at 5 cm above the surface of liquid nitrogen for 5 min,and finally preserve in liquid nitrogen.By this procedure,the motility of frozen-thawed sperm could reach (83±3) % and the hatching rate after artificial insemination was about 70%,which can meet the requirement of lager scale breeding of T. bimaculatus. And our result could also be a good reference for relative researches in other fugu fishes.