试验对西伯利亚鲟、美洲鲥鱼和暗纹东方鲀3种长江珍稀鱼类的精液进行超低温冷冻保存技术研究。以解冻后精子的活力为参数，探讨了稀释液、不同种类不同体积分数的保护剂、以及不同的精液稀释比例对实验鱼类精液进行超低温冷冻保存的保护效果。结果显示，3种长江珍稀鱼类以配方2和体积分数为10%甲醇组成抗冻保护剂，西伯利亚鲟精液按1:2稀释倍数，保存10天后检查精子活力，存活率可达(45±5.00)%。暗纹东方鲀精液按1:2稀释倍数，保存20天后检查精子活力，存活率可达(60.5±2.69)%。美洲鲥鱼精液按1:3稀释倍数，保存60天后检查精子活力，存活率可达(80.3±2.58)%。3种珍稀鱼类的精子保存以美洲鲥鱼效果最好。保存方法均采用三步法，在液氮面上方6 cm处平衡10 min，接着在液氮面上平衡5 min，最后投入液氮中冷冻保存。研究结果为保护长江珍稀鱼类的种质资源提供了理论基础。
Abstract
The technique of sperm cryopreservation of three rare migratory fishes in the Yangtze River was studied, including Acipenser baerii, Alosa sapidissima and Takifugu obscurus. The success of sperm cryopreservation was evaluated by post-thaw sperm motility. To explore methods for sperm cryopreservation, this study evaluated the effects of different extenders, cryoprotectants, their volume fractions, equilibration time and dilution ratios. The result showed that the best sperm cryopreservation of motility of Acipenser baerii was (45±5.00)% and Takifugu obscurus (60.5±2.69)% were obtained by following procedure: the ringer solution and the semen were mixed at ratio of 2:1 in volume and added 10% methanol, while the best sperm cryopreservation of motility of Alosa sapidissima (80.3±2.58)% was obtained by following procedure: the ringer solution and the semen were mixed at ratio of 3:1 in volume and added 10% methanol. Sperms were cryopreserved by three-step cooling progam. These results provided the methods to conserve genetic resources in three rare migratory fishes in the Yangtze River.
关键词
西伯利亚鲟，美洲鲥鱼，暗纹东方鲀，精液，超低温保存
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Key words
Acipenser baerii; Alosa sapidissima; Takifugu obscurus; sperm; cryopreservation
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参考文献
[1] Smith Au, Polge C. Survival of spermatozoa at low temperatures[J]. Nature,1950, 166: 668-669.
[2] 罗刚,高勇,陈学洲,等.北方池塘西伯利亚鲟养殖试验[J].中国水产,2008(10):43-44.
[3] 杨移斌,夏永涛,曹海鹏,等.西伯利亚鲟基本生物学特性及养殖介绍[J].水产养殖,2013(11):17-19.
[4] 刘鹏,庄平,章龙珍,等.人工养殖西伯利亚鲟精子超低温冷冻保存研究[J].海洋科学,2007,29(2):120-127.
[5] 徐钢春,张呈祥,郑金良,等.美洲鲥的人工繁殖及胚胎发育的研究[J].海洋科学,2012,36(7):89-96.
[6] 赵强、赵清良,殷宁.暗纹东方鲀精液冷藏对精子活力、受精率和孵化率的影响[J].南京农业大学学报,1999,22(4):57-60.
[7] Pan J L, Ding S Y, Ge J C. Development of cryopreservation for maintainingyellow catfishPelteobagrus fulvidraco sperm[J]. Aquaculture, 2008,279(1/2/3/4):173 -176.
[8] 夏良萍,陈梦婷,陈悦萍,等.黄颡鱼精子超低温冷冻保存技术[J].江苏农业科学,2013,41(10):196-198.
[9] 王晓爱,杨君兴,陈小勇,等.4种渗透性抗冻剂对暗色唇鱼精子冷冻保存的影响[J].水生态学杂志,2012(5):88-93.
[10] 洪万树,张其永,许胜发,等.花鲈精子生理特性及其精液超低温冷冻保存[J]海洋学报,1996,18(2):97-104.
[11] 林丹军,尤永隆.大黄鱼精子生理特性及其冷冻保存[J].热带海洋学报,2002,21(4):69-75.
[12] 于海涛,张秀梅,陈超,等.鱼类精液超低温冷冻保存的研究展望[J].海洋湖沼通报,2004(2):66-71.
[13] Li J, Li Y, Zhao X X, et al. Sperm cells as vectors for introducing foreign DNA into eggs of Tilapoa[J]. Biotechnology,1994(4):20-22.
[14] Paniagua-Chavez CG, Buchanan JT, Tiersch TR. 1998. Effect of extendersolutions and dilution on motility and fertilizing ability of Easternoyster sperm[J]. J Shellfish Res,17(1-3):231-237
[15] Tiersch T, Yang H, Jenkins JA, Dong Q. 2007. Sperm cryopreservation in fish and shellfish[J]. Soc Reprod Fertil Suppl,65(sup):493-508.
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