| 摘要: |
| 为建立点带石斑鱼(Epinephelus malabaricus)精子冷冻保存技术, 保存点带石斑鱼种质资源, 作者通过选用TS-19、TS-2、Hank’s、MPRS、Cortland、0.75% NaCl 等作为稀释液及10%～25% 二甲亚砜、甘油、甲醇作为抗冻液, 对繁殖季节中3 个不同时期的点带石斑鱼精液进行超低温冷冻保存研究。研究结果表明: (1) Hank’s 溶液为点带石斑鱼精子冷冻保存最适冷冻稀释液; (2) Hank’s 溶液与10%～15% DMSO 溶液为点带石斑鱼精子保存的最适冷冻保护液组合, 可以获得60%以上的冻精成活率, 而使用不同浓度的MeOH 作为抗冻液时, 冻精成活率均为0%; (3) 以10%～25% Gly 作为抗冻液时,冷冻前未加天然海水时均能激活点带石斑鱼精子正常运动, 其效果与天然海水相似, 加入天然海水后精子运动效果急剧下降, 精子冷冻保存后也存在同样现象; (4) 为保证获得更高的冻精成活率, 点带石斑鱼精液冷冻保存样品的最佳获取时期是在亲鱼自然繁殖期前进行为宜。 |
| 关键词: 点带石斑鱼(Epinephelus malabaricus) 精子 超低温冷冻保存 稀释液 抗冻液 |
| DOI：10.11759/hykx20130809001 |
| 分类号: |
| 基金项目:国家“863”计划资助项目(2012AA10A407); 海南省自然科学基金资助项目(313046); 海南大学青年基金资助项目(qnjj1154) |
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| The cryopreservation of spermatozoa of Epinephelus malabaricus |
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| Abstract: |
| Epinephelus malabaricus is an important mariculture species in southern China. The spermatoza cryopreservation is important for artificial breeding, cross-breeding and germplasm pool establishment. In this paper, six different extender solutions (TS-19, TS-2, Hank’s, MPRS, Cortland and 0.75% NaCl) were chosen as diluents, and three (10%～25% dimethyl sulphoxide, glycerol, methanol) as cryoprotectants for the cryopreservation of spermatozoa of E. malabaricus. The results show: (1) Compared with other diluents, the Hank’s solution was the optimum one for the cryopreservation of spermatozoa of E. malabaricus; (2) The optimum combination for the cryopreservation of spermatozoa of E. malabaricus was the combination of Hank’s solution with 10%～15% DMSO solution, which can reach a survival rate of more than 60%, while any concentration of MeOH resulted in a survival rate of 0%; (3) When the 10%～25% glycerol solution was chosen as cryoprotectant, the spermatozoa of E. malabaricus could keep normal activity, and the activity was similar to that in natural seawater before the cryopreservation. But the spermatozoa motility decreased rapidly when seawater was added to cryoprotectant. Similar phenomenon existed after the cryopreservation; (4) To gain a better survival rate of frozen-thawed semen, the optimal collection period of cryopreservation semen sampling of E. malabaricus was 1 day before the spermiation. |
| Key words: Epinephelus malabaricus spermatozoa cryopreservation extender solution cryoprotectant |
