太平洋牡蛎精液的超低温保存

Cryopreservation of Pacific Oyster (Crassostrea gigas) Spermatoza

利用解剖法采集太平洋牡蛎精液 ,用自然海水做基础溶液 ,配制不同浓度的 DMSO(二甲亚砜 )作为超低温保护剂 ,在液氮 (L N2 )面上不同高度进行预处理 ,投入 LN2 内保存 ,升温解冻后 ,检测精子成活率。结果表明 ,DMSO处理浓度、预处理温度及时间和升温方式对超低温保存成活率均具有显著影响。其中 ,用 10 % DMSO在 LN2 面上 17cm预处理 6 min,采用 16～ 17℃流水不完全解冻 ,效果最好 ,超低温保存精子成活率可达 70 %。与新鲜精子相比 ,在 L N2 内保存一天的精子 ,授精能力没有明显变化。

The spermatozoa were obtained from the Pacific oyster gonad by dissection. The dimethyl sulphoxyde(DMSO) melted by filtrated sea water was used as cryoprotectant. Prefreezing was conducted by balancing over different heights from the surface of liquid nitrogen (LN 2),and thaw took place after immersion into LN 2for two hours. The result indicated that the concentration of DMSO and the temperature and time of prefreezing and the thawing method were related to the survival rate of cryopreserved spermatozoa. And it was found that the survival rate reached 70% when prefreezing 6min at 17cm over the surface of LN 2, treating with 10% DMSO and thawing with 16～17℃ flowing sea water, The fertilization rate and hatchery rate reached 68% and 64%, respectively.