Department for Organismic Biology, University of Salzburg, Salzburg, Austria.
Theriogenology. 2011 Sep 15;76(5):882-90. doi: 10.1016/j.theriogenology.2011.04.019. Epub 2011 Jun 25.
Until now the supplementation of cryopreservation extenders with antioxidants has not been examined in teleost fish. Therefore, the present study investigated whether addition of antioxidants (catalase, superoxide dismutase, peroxidase, reduced glutathione, reduced methione, mixtures of reduced and oxidized glutathione or methionine) to the cryopreservation extenders could increase the quality of frozen-thawed semen of brook trout, Salvelinus fontinalis, and rainbow trout, Oncorhynchus mykiss. In brook trout and rainbow trout semen post-thaw fertility and motility were evaluated and in brook trout additionally the membrane integrity, DNA integrity, and sperm lipid peroxidation were evaluated. The tested antioxidants affected the motility parameters, DNA integrity, and fertility of cryopreserved semen, but not the membrane integrity. Most of the observed effects were negative and only minor positive effects were found. In brook trout 1.5 mmol/l reduced methionine and a mixture of 1.5 mmol/l oxidized and reduced glutathione increased the swimming velocity of frozen-thawed semen. One hundred U/l catalase, 1.5 mmol/l reduced glutathione, and 1.5 mmol/l reduced methionine slightly, but not statistically significantly increased the semen post-thaw fertility. However, these effects were not detectable in rainbow trout. Antioxidative stress or damage seems to play no role during cryopreservation, as also in the lipid peroxidation test no differences were obtained between fresh and cryopreserved semen. Therefore, for routine cryopreservation extender supplementation with antioxidants is not recommended in brook trout and rainbow trout.
到目前为止,抗氧化剂在冷冻保护剂中的添加在硬骨鱼类中还没有被研究过。因此,本研究调查了在冷冻保护剂中添加抗氧化剂(过氧化氢酶、超氧化物歧化酶、过氧化物酶、还原型谷胱甘肽、还原型蛋氨酸、还原型和氧化型谷胱甘肽混合物或蛋氨酸)是否能提高斑点叉尾鮰和虹鳟的冷冻-解冻精液的质量。在斑点叉尾鮰和虹鳟的精液中,评估了冷冻-解冻后的活力和运动能力,而在斑点叉尾鮰中,还评估了膜完整性、DNA 完整性和精子脂质过氧化。测试的抗氧化剂影响了冷冻保存精液的运动参数、DNA 完整性和活力,但不影响膜完整性。大多数观察到的影响是负面的,只有少数是正面的。在斑点叉尾鮰中,1.5mmol/L 的还原型蛋氨酸和 1.5mmol/L 的氧化型和还原型谷胱甘肽混合物增加了冷冻-解冻精液的游泳速度。100U/L 的过氧化氢酶、1.5mmol/L 的还原型谷胱甘肽和 1.5mmol/L 的还原型蛋氨酸略微但无统计学意义地提高了精液解冻后的活力。然而,这些效果在虹鳟中无法检测到。抗氧化应激或损伤在冷冻保存过程中似乎没有发挥作用,因为在脂质过氧化试验中,新鲜和冷冻保存的精液之间也没有差异。因此,对于常规的冷冻保护剂,不建议在斑点叉尾鮰和虹鳟中添加抗氧化剂。