冷冻保存鲤鱼精子的生育能力

M. Sultana, M. Nahiduzzaman, Mahbubul Hassan, M. Khanam, M. Hossain
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引用次数: 13

摘要

本研究旨在规范鲤鱼精子的冷冻保存方案,以供人工繁殖之用。三种填充剂(Alsever溶液、柠檬酸蛋黄和0.9% NaCl)与三种低温保护剂(乙醇、甲醇和DMSO)混合。两步冷冻方案(5°C至-4°C,速度为4°C /分钟,然后-4°C至-80°C,速度为10°C /分钟,最后保存10分钟)在计算机控制的冷冻室中进行,其中0.25 ml含有精子的吸管储存在液氮容器中,温度为-196°C。在冷冻前(分别为93.33±1.05、90±1.83和88.33±2.11)和解冻后(分别为80±4.28、74.17±3.96和76.67±4.59),10%的冷冻保护剂在解冻后的运动性最好。综合考虑鲤鱼精子冷冻前和解冻后的活力,低温和低温稀释剂的最佳配比为1:9。与其他组合相比,柠檬酸盐添加DMSO的受精率(37.67±1.76%)和孵化率(28.67±1.85%)最高。关键词:鲤鱼;精子;低温贮藏;冷冻保护剂;cryodiluent;生育能力。DOI: 10.3329/ujzru.v28i0.5287 Univ. j. zool。Rajshahi university . Vol. 28, 2010 pp. 51-55
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Fertility of cryopreserved common carp ( Cyprinus carpio ) spermatozoa
The study was aimed at standardization of cryopreservation protocol for the common carp, ( Cyprinus carpio ) spermatozoa for using in the artificial propagation. Three extenders (Alsever’s solution, egg-yolk citrate and 0.9% NaCl) were combined with three cryoprotectants (ethanol, methanol and DMSO). Two-step freezing protocol (5°C to -4°C at a rate of 4°C per minute then -4°C to -80°C at a rate of 10°C per minute and finally held for 10 min) was carried out in a computer-controlled freezer where 0.25 ml straws containing spermatozoa were stored in a liquid nitrogen container at -196°C. Alsever’s solution with ethanol, Alsever’s solution with methanol and egg yolk citrate with DMSO gave better motility (%) in both pre-freeze (93.33±1.05, 90±1.83 and 88.33±2.11, respectively) and post-thaw (80±4.28, 74.17±3.96 and 76.67±4.59, respectively) periods whereas, 10% cryoprotectant gave the best motility during post-thaw period. Considering both pre-freeze and post-thaw motilities of the common carp spermatozoa, 1:9 was the best ratio for the milt and cryodiluents. The egg yolk citrate with DMSO yielded the best fertilization (37.67±1.76%) and hatching (28.67±1.85%) compared to other combinations under study. Key words: Cyprinus carpio ; spermatozoa; cryopreservation; cryoprotectant; cryodiluent; fertility. DOI: 10.3329/ujzru.v28i0.5287 Univ. j. zool. Rajshahi Univ. Vol. 28, 2010 pp. 51-55
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