软鳍新光唇鱼精子的超低温冷冻保存

2011年,对软鳍新光唇鱼(Neolissochilus benasi)进行了精子超低温冷冻保存研究。以解冻后的精子活力为指标,采用稀释液D-15,设计不同的抗冻剂种类和浓度,以及不同的实验条件(包括冷冻体积、4℃平衡时间和解冻温度等)探索软鳍新光唇鱼精子的超低温冷冻保存方法。筛选出了适合软鳍新光唇鱼精子超低温冷冻保存的两种抗冻保护剂及其浓度分别为10%MeOH和15%EG,确定了精液与稀释液的最适稀释比例为1:7、4℃平衡时间区间为10～60min、冷冻体积为60μL,以及复苏方法为37℃水浴快速解冻30s。当鲜精活力为(62.33±2.05)%,综合以上最佳实验条件进行保存,解冻后精子的最高活力为(29.67±0.47)%,但效果不理想,不能达到广泛生产运用水平;产生这一结果,可能与异地保育物种的饲养管理有关。因此,在亲鱼培育管理中要最大限度地降低捕获诱发的压力,尽量提供适合的养殖条件。在珍稀鱼类异地保育时,繁殖用雄鱼的培育与雌鱼同等重要,是获得大量高质量仔鱼的关键。

Cryopreservation of sperm from Neolissochilus benasi

Cryopreservation of sperm from Neolissochilus benasi was studied in 2011. The effects of various cryoprotectants of different concentrations, dilution ratios of milt to extender, storage volume and thawing temperature on motility of post-thawing of spermatozoa were examined to optimize cryopreservation procedures. Semen was stored in liquid nitrogen in 1.8 mL cryovial for 24 h, and the intensity of sperm motility was measured before and after cryopreservation. Post-thawing motility of frozen sperm obtained with cryoprotectants 10% MeOH or 15% EG were higher than for others. The most effective dilution ratio of milt to extender is 1:7. The maximal storage volume is 60 μL of 1.8 mL cryovial and the optimal sperm equilibration period in the extender D-15+10% MeOH was between 10-60 min. Thawing was optimal in a 37 Degrees Celsius water bath. When fresh sperm motility is (62.33±2.05)%, this cryopreservation protocol resulted in frozen-thawed semen with 20%-30% motile. The overall effect is not ideal, and cannot achieve extensive application. Different breeding management of different ground protection may have contributed to this result. Therefore, it is necessary to reduce stress capture induced in management of parent fish and provide suitable forming conditions. In the ex situ conservation of rare fish the broodstocks management of males is as important as that for females and the key to obtaining high quality larval fish.