Wang Zhicheng, Gu Haotian, Zhu Chunhong, Wang Yifei, Liu Hongxiang, Song Weitao, Tao Zhiyun, Xu Wenjuan, Zhang Shuangjie, Li Huifang
Jiangsu Institute of Poultry Science, Yangzhou 225125, China.
Animals (Basel). 2025 Aug 6;15(15):2309. doi: 10.3390/ani15152309.
Waterfowl semen cryopreservation technology is a key link in genetic resource conservation and artificial breeding, but poultry spermatozoa, due to their unique morphology and biochemical properties, are prone to oxidative stress during freezing, resulting in a significant decrease in vitality. In this study, we first used four different freezing procedures (P1-P4) to freeze duck semen and compared their effects on duck sperm quality. Then, the changes in antioxidant indexes in semen were monitored. The results showed that program P4 (initial 7 °C/min slow descent to -35 °C, followed by 60 °C/min rapid descent to -140 °C) was significantly better than the other programs ( < 0.05), and its post-freezing sperm vitality reached 71.41%, and the sperm motility was 51.73%. In the P1 and P3 groups, the sperm vitality was 65.56% and 53.41%, and the sperm motility was 46.99% and 31.76%, respectively. In terms of antioxidant indexes, compared with the fresh semen group (CK), the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-px) in the P2 group were significantly decreased ( < 0.05), while the activities of SOD and CAT in the P4 group showed no significant changes ( > 0.05) except that the activity of GSH-px was significantly decreased ( < 0.05). And the CAT and GSH-px activities in the P4 group were significantly higher than those in the P2 group ( < 0.05). The content of malondialdehyde (MDA) in the P2 group was significantly higher than that in the fresh semen group ( < 0.05), and there was no significant difference between the P2 group and the P4 group ( > 0.05). The total antioxidant capacity (T-AOC) content of the P2 and P4 groups was significantly lower than that of the fresh semen group ( < 0.05). The staged cooling strategy of P4 was effective in reducing the exposure time to the hypertonic environment by balancing intracellular dehydration and ice crystal inhibition, shortening the reactive oxygen species accumulation and alleviating oxidative stress injury. On the contrary, the multi-stage slow-down strategy of P2 exacerbated mitochondrial dysfunction and the oxidative stress cascade response due to prolonged cryogenic exposure time. The present study confirmed that the freezing procedure directly affects duck sperm quality by modulating the oxidative stress pathway and provides a theoretical basis for the standardization of duck semen cryopreservation technology.
水禽精液冷冻保存技术是遗传资源保护和人工繁育的关键环节，但家禽精子由于其独特的形态和生化特性，在冷冻过程中易遭受氧化应激，导致活力显著下降。在本研究中，我们首先采用四种不同的冷冻程序（P1 - P4）冷冻鸭精液，并比较它们对鸭精子质量的影响。然后，监测精液中抗氧化指标的变化。结果表明，程序P4（初始以7℃/min缓慢降至 - 35℃，随后以60℃/min快速降至 - 140℃）明显优于其他程序（P < 0.05），其冷冻后精子活力达到71.41%，精子运动率为51.73%。在P1和P3组中，精子活力分别为65.56%和53.41%，精子运动率分别为46.99%和31.76%。在抗氧化指标方面，与新鲜精液组（CK）相比，P2组中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GSH - px）的活性显著降低（P < 0.05），而P4组中除GSH - px活性显著降低（P < 0.05）外，SOD和CAT的活性无显著变化（P > 0.05）。并且P4组中的CAT和GSH - px活性显著高于P2组（P < 0.05）。P2组中丙二醛（MDA）的含量显著高于新鲜精液组（P < 0.05），P2组和P4组之间无显著差异（P > 0.05）。P2组和P4组的总抗氧化能力（T - AOC）含量均显著低于新鲜精液组（P < 0.05）。P4的分段降温策略通过平衡细胞内脱水和冰晶抑制，有效减少了高渗环境暴露时间，缩短了活性氧积累并减轻了氧化应激损伤。相反，P2的多阶段减速策略由于延长了低温暴露时间，加剧了线粒体功能障碍和氧化应激级联反应。本研究证实冷冻程序通过调节氧化应激途径直接影响鸭精子质量，并为鸭精液冷冻保存技术的标准化提供了理论依据。
