Optimization of Duck Semen Freezing Procedure and Regulation of Oxidative Stress

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 t...

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Main Authors: Zhicheng Wang, Haotian Gu, Chunhong Zhu, Yifei Wang, Hongxiang Liu, Weitao Song, Zhiyun Tao, Wenjuan Xu, Shuangjie Zhang, Huifang Li
Format: Article
Language:English
Published: MDPI AG 2025-08-01
Series:Animals
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Online Access:https://www.mdpi.com/2076-2615/15/15/2309
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author Zhicheng Wang
Haotian Gu
Chunhong Zhu
Yifei Wang
Hongxiang Liu
Weitao Song
Zhiyun Tao
Wenjuan Xu
Shuangjie Zhang
Huifang Li
author_facet Zhicheng Wang
Haotian Gu
Chunhong Zhu
Yifei Wang
Hongxiang Liu
Weitao Song
Zhiyun Tao
Wenjuan Xu
Shuangjie Zhang
Huifang Li
author_sort Zhicheng Wang
collection DOAJ
description 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 (<i>p</i> < 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 (<i>p</i> < 0.05), while the activities of SOD and CAT in the P4 group showed no significant changes (<i>p</i> > 0.05) except that the activity of GSH-px was significantly decreased (<i>p</i> < 0.05). And the CAT and GSH-px activities in the P4 group were significantly higher than those in the P2 group (<i>p</i> < 0.05). The content of malondialdehyde (MDA) in the P2 group was significantly higher than that in the fresh semen group (<i>p</i> < 0.05), and there was no significant difference between the P2 group and the P4 group (<i>p</i> > 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 (<i>p</i> < 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.
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spelling doaj-art-8e71c3a65cf6429496ceafbb35e08f102025-08-20T04:00:54ZengMDPI AGAnimals2076-26152025-08-011515230910.3390/ani15152309Optimization of Duck Semen Freezing Procedure and Regulation of Oxidative StressZhicheng Wang0Haotian Gu1Chunhong Zhu2Yifei Wang3Hongxiang Liu4Weitao Song5Zhiyun Tao6Wenjuan Xu7Shuangjie Zhang8Huifang Li9Jiangsu Institute of Poultry Science, Yangzhou 225125, ChinaJiangsu Institute of Poultry Science, Yangzhou 225125, ChinaJiangsu Institute of Poultry Science, Yangzhou 225125, ChinaJiangsu Institute of Poultry Science, Yangzhou 225125, ChinaJiangsu Institute of Poultry Science, Yangzhou 225125, ChinaJiangsu Institute of Poultry Science, Yangzhou 225125, ChinaJiangsu Institute of Poultry Science, Yangzhou 225125, ChinaJiangsu Institute of Poultry Science, Yangzhou 225125, ChinaJiangsu Institute of Poultry Science, Yangzhou 225125, ChinaJiangsu Institute of Poultry Science, Yangzhou 225125, ChinaWaterfowl 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 (<i>p</i> < 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 (<i>p</i> < 0.05), while the activities of SOD and CAT in the P4 group showed no significant changes (<i>p</i> > 0.05) except that the activity of GSH-px was significantly decreased (<i>p</i> < 0.05). And the CAT and GSH-px activities in the P4 group were significantly higher than those in the P2 group (<i>p</i> < 0.05). The content of malondialdehyde (MDA) in the P2 group was significantly higher than that in the fresh semen group (<i>p</i> < 0.05), and there was no significant difference between the P2 group and the P4 group (<i>p</i> > 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 (<i>p</i> < 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.https://www.mdpi.com/2076-2615/15/15/2309Jinding duckssemen freezing proceduresemen qualityoxidative stresslipid peroxidation
spellingShingle Zhicheng Wang
Haotian Gu
Chunhong Zhu
Yifei Wang
Hongxiang Liu
Weitao Song
Zhiyun Tao
Wenjuan Xu
Shuangjie Zhang
Huifang Li
Optimization of Duck Semen Freezing Procedure and Regulation of Oxidative Stress
Animals
Jinding ducks
semen freezing procedure
semen quality
oxidative stress
lipid peroxidation
title Optimization of Duck Semen Freezing Procedure and Regulation of Oxidative Stress
title_full Optimization of Duck Semen Freezing Procedure and Regulation of Oxidative Stress
title_fullStr Optimization of Duck Semen Freezing Procedure and Regulation of Oxidative Stress
title_full_unstemmed Optimization of Duck Semen Freezing Procedure and Regulation of Oxidative Stress
title_short Optimization of Duck Semen Freezing Procedure and Regulation of Oxidative Stress
title_sort optimization of duck semen freezing procedure and regulation of oxidative stress
topic Jinding ducks
semen freezing procedure
semen quality
oxidative stress
lipid peroxidation
url https://www.mdpi.com/2076-2615/15/15/2309
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