Department of Fisheries, Graduate School of Agriculture, Kinki University, Naka-Machi, Nara 631-8505, Japan.
Cryobiology. 2010 Dec;61(3):263-7. doi: 10.1016/j.cryobiol.2010.09.003. Epub 2010 Sep 29.
In the present study, we examined methods for the cryopreservation of Epinephelus septemfasciatus spermatozoa. The percent motility, average path velocity, and linearity of movement (LIN) of fresh and corresponding post-thaw sperm were evaluated. Sperm motility was investigated using computer-assisted sperm analysis. Five percent dimethyl sulphoxide (Me₂SO) with 95% fetal bovine serum (FBS) was the most successful cryoprotectant diluent with a comparative post-thaw motility of 77.6±8.5%; 5% dimethyl formamide was also effective. Fetal bovine serum was significantly better as an extender when compared with artificial seminal plasma, glucose, and trehalose solution. Sperm tolerated a wide range of cooling rates (from 27.1 to 94.3 °C min⁻¹); however, the post-thaw motility of sperm cooled to -30 °C was significantly lower than that of other cooled temperatures (-40 to -70 °C). The velocity of post-thaw sperm was significantly lower than that of fresh sperm, although LIN remained the same. For effective cryopreservation of seven-band grouper sperm, samples should be diluted in 5% Me₂SO with 95% FBS and cooled to at least -40 °C before immersion in liquid nitrogen.
在本研究中，我们研究了斜带石斑鱼精子的冷冻保存方法。评估了新鲜和相应解冻后精子的运动百分率、平均路径速度和运动直线性（LIN）。使用计算机辅助精子分析研究精子活力。5%二甲基亚砜（Me₂SO）加 95%胎牛血清（FBS）是最成功的冷冻保护剂稀释剂，解冻后活力为 77.6±8.5%；5%二甲基甲酰胺也很有效。与人工精液、葡萄糖和海藻糖溶液相比，胎牛血清作为稀释液显著更好。精子能够耐受广泛的冷却速率（从 27.1 到 94.3°C min⁻¹）；然而，冷却至-30°C 的解冻后精子活力明显低于其他冷却温度（-40 至-70°C）。解冻后精子的速度明显低于新鲜精子，尽管 LIN 保持不变。为了有效冷冻保存七带石斑鱼精子，应将样品稀释在 5%Me₂SO 和 95%FBS 中，并冷却至至少-40°C，然后再浸入液氮中。
