Short-term storage of semen is a necessary key procedure in fish; it allows maximizing the use of gametes. Nevertheless, sperm quality decreases during storage has been associated with oxidative stress damage due to an increase in reactive oxygen species (ROS) during storage. This study was designed to optimize a short-term storage protocol for Coho salmon (Oncorhynchus kisutch) spermatozoa, evaluating the effect of extender dilution and the addition of butyl-hydroxytoluene (BHT) antioxidant on sperm function parameters. In the first experiment, fresh semen was diluted in Storfish®: extender dilution (1:2 and 1:3) and a control sample undiluted and stored at 4 °C for 7-days. In both experiments motility (MO), viability and integrity of plasma membrane, mitochondrial membrane potential (MMP) and superoxide anion level (O2-) were evaluated at 0, 3 and 7 days. Result shows that, 1:3 dilution maintained a higher sperm function for a longer period time. In the second experiment, spermatozoa were suspended in Storfish® (1:3) supplemented with two different concentrations of BHT (1.0 mM and 2.0 mM) and a control sample without antioxidant and stored at 4 °C for 7 days. The results demonstrated that, antioxidant-supplemented samples greater MO than control samples (P < 0.05). The viability remained >75% during storage in all groups. MMP was higher in 2.0 mM BHT compared to 1.0 mM and control (P < 0.05), in addition, this concentration reduced O2- level (P < 0.05). In conclusion, sperm: extender dilution 1:3 and adding of 2.0 mM BHT in sperm storage extender may enhance protection sperm function in Oncorhynchus kisutch against effects harmful of the oxidative stress during the in vitro storage.

中文翻译：

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银大麻哈鱼 (Oncorhynchus kisutch) 在 4 °C 下的短期储存精子：精子的影响：稀释剂稀释比和抗氧化剂丁基羟基甲苯 (BHT) 对精子功能的影响

精液的短期储存是鱼类必要的关键程序；它允许最大限度地利用配子。然而，由于储存过程中活性氧 (ROS) 的增加，储存过程中精子质量的下降与氧化应激损伤有关。本研究旨在优化银鲑鱼 (Oncorhynchus kisutch) 精子的短期储存方案，评估稀释剂稀释和添加丁基羟基甲苯 (BHT) 抗氧化剂对精子功能参数的影响。在第一个实验中，新鲜精液在 Storfish®：稀释剂稀释液（1:2 和 1:3）中稀释，对照样品未稀释并在 4 °C 下储存 7 天。在这两个实验中，质膜的活力 (MO)、活力和完整性、线粒体膜电位 (MMP) 和超氧阴离子水平 (O2-) 均在 0、3天和7天。结果表明，1:3 的稀释度可以在更长的时间内保持较高的精子功能。在第二个实验中，精子悬浮在 Storfish® (1:3) 中，添加了两种不同浓度的 BHT（1.0 mM 和 2.0 mM）和不含抗氧化剂的对照样品，并在 4 °C 下储存 7 天。结果表明，添加抗氧化剂的样品的 MO 大于对照样品（P < 0.05）。在所有组中，在储存期间存活率保持>75%。与 1.0 mM 和对照相比，2.0 mM BHT 中的 MMP 更高（P < 0.05），此外，该浓度降低了 O2 水平（P < 0.05）。总之，精子：稀释剂稀释 1:3 并添加 2。