人工养殖西伯利亚鲟精子超低温冷冻保存研究

研究了人工养殖西伯利亚鲟精子的生物学特征及超低温冷冻保存方法。西伯利亚鲟的产精量为113.67±39.86 ml,精子密度为(6.49±3.10)×108/ml,精子活力为(85.4±9.5)%,精子寿命为353±23 s。精子密度与精子快速运动时间、精子寿命之间均存在线性相关,用方程分别表示为:y=1.0384x+1.5089(R2=0.7325);y=2.9069x+74.289(R2=0.6967)。结果表明精子密度可作为一项精子质量评价的标准。通过比较西伯利亚鲟精子在不同稀释液、不同抗冻剂和抗冻剂浓度、降温速率、解冻温度下的保存效果,结果表明:配方2作为稀释液,18%甲醇作为抗冻剂,二步法超低温(-196℃)冷冻保存精子,40℃水浴解冻取得最好的冻后活力,解冻后活力为(51.8±5.8)%。西伯利亚鲟授精的最佳精卵比为106∶1。在此精卵比下用冻精授精分别得到了(72.3±3)%的受精率和(52.9±4.1)%的孵化率,其中受精率与鲜精没有显著性差异,孵化率与鲜精有显著差异(P<0.05)。

Study on cryopreservation of spermatozoa in cultured Siberian sturgeon (Acipenser baerii baerii)

The quantity of sperm for each mature cultured Siberian sturgeon was 113.67±39.86 ml,sperm concentration was(6.49±3.10)×108/ml,sperm motility was(85.4±9.5)% and life-span was 353±23 s.Sperm concentration was linear with fast-moving period and life-span,and it could be described by the equation respectively:!y=1.038 4x+1.508 9(R2 = 0.732 5);y = 2.906 9x+74.289(R2 = 0.696 7).Therefore,we may use sperm concentration as a standard to evaluate the quality of Siberian sturgeon sperm.We used different methods to preserve the sperm,such as,different extender,cryoprotectants and its concentration,freezing rate,thawing rate,etc..The results indicated that using Tris-Glu-KCl supplemented with 18% methanol as extender,two steps freezing procedure and 40 ℃ thawing water bath temperature was the best method (with motility of(51.8±5.8%)]for the cryopreservation of Siberian sturgeon semen.For the best fertilization,the ratio of sperm to egg should be 106: 1.Using this ratio of thawed sperm and egg,fertilizing rate (72.3±3)%]and hatching rate (52.9±4.1)%]might be obtained.There was no significant difference on fertilizing rate between fresh and thawed semen,however,there was significant difference on hatching rate between fresh and thawed semen(P < 0.05).