Effect of oxygen and antioxidants on the common carp (Cyprinus carpio L.) milt during short-term storage

IF 3.9 1区农林科学 Q1 FISHERIES

Aquaculture Pub Date : 2025-09-11 DOI:10.1016/j.aquaculture.2025.743172

Agnieszka Brzyszcz , Beata Sarosiek , Katarzyna Dryl , Katarzyna Targońska , Michał Blitek , Dariusz Kucharczyk , Wojciech Skorupa , Masaya Morita , Radosław K. Kowalski

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Abstract

This study explores the impact of oxygen and antioxidants in an immobilizing buffer on short-term storage of common carp sperm. Sperm from five carp individuals underwent short-term storage for 10 days without dilution (Control), mixed with buffer TLP alone (TLP), or with antioxidants: 5 mM glutathione (Glut), 5 mM ascorbic acid (VitC), 1.5 mg/mL vitamin E (VitE), and a mix of all mentioned antioxidants (Mix). Diluted milt samples were stored at 4 °C in either open Eppendorf tubes within oxygen-filled bags (refreshed every 24 h) or closed tubes under limited aerobic conditions. All samples were gently mixed twice daily to prevent spermatozoa sedimentation. Computer-assisted sperm analysis (CASA) was employed to measure the curvilinear velocity (VCL) and total motility (MOT) of stored samples. Live/Dead double staining kit assessed sperm membrane integrity. After 10 days of storage, a fertility trial was conducted to compare preserved milt with freshly collected sperm samples (n = 3). In samples stored in limited aerobic conditions, the VCL parameter in TLP, Glut, and VitE buffers remained steady. Oxygen altered MOT and VCL, with the best variants of diluents (TLP and VitE) showing a decrease from 90 % to about 80 %. In samples with the addition of ascorbic acid and the antioxidant mixture, a loss of viability was observed after 6 days. On the other hand, membrane integrity in limited aerobic conditions (in buffers TLP, Glut, and VitE) remained unaffected to day 10. Fertilization success, measured by hatching rate, indicated that diluted common carp milt, stored for 10 days in aerobic conditions, maintained its fertilizing potential. This study demonstrates the impact of oxygen utilization and the potential beneficial effect of antioxidant supplementation on the short-term storage of common carp milt in aquaculture practices. Correlation analysis conducted in this study revealed strong positive relationships between hatching rate and motility parameters, such as ALH and total motility, highlighting the importance of these factors in successful common carp fertilization.

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氧和抗氧化剂对鲤鱼短期贮藏的影响

本研究探讨了固定缓冲液中氧和抗氧化剂对鲤鱼精子短期储存的影响。5条鲤鱼的精子在不稀释的情况下短期储存10天（对照组），单独与缓冲TLP （TLP）混合，或与抗氧化剂混合：5 mM谷胱甘肽（Glut）， 5 mM抗坏血酸（VitC）， 1.5 mg/mL维生素E (VitE)，以及所有上述抗氧化剂的混合物（mix）。稀释后的牛奶样品在4°C下保存在开放的Eppendorf管中，装在充氧袋中（每24小时更新一次），或在有限的有氧条件下保存在封闭的管中。所有样本每天温和混合两次，以防止精子沉降。采用计算机辅助精子分析（CASA）方法测定存储样品的曲线速度（VCL）和总运动力（MOT）。活/死双染色试剂盒评估精子膜完整性。保存10天后，进行生育试验，比较保存的牛奶和新鲜收集的精子样本（n = 3）。在有限有氧条件下储存的样品中，TLP、Glut和VitE缓冲液中的VCL参数保持稳定。氧改变了MOT和VCL，稀释剂（TLP和VitE）的最佳变体显示从90%下降到约80%。在加入抗坏血酸和抗氧化剂混合物的样品中，6天后观察到活力丧失。另一方面，在有限的有氧条件下（在缓冲液中TLP、Glut和VitE），膜的完整性在第10天仍未受到影响。以孵化率衡量的受精成功率表明，稀释后的鲤鱼粉在有氧条件下保存10天，仍能保持其受精潜力。本研究探讨了养殖过程中氧利用的影响以及添加抗氧化剂对鲤鱼鱼苗短期贮藏的潜在有益影响。本研究进行的相关分析显示，孵化率与运动参数（如ALH和总运动）之间存在很强的正相关关系，突出了这些因素对鲤鱼成功受精的重要性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Aquaculture 农林科学-海洋与淡水生物学

CiteScore

8.60

自引率

17.80%

发文量

1246

审稿时长

56 days

期刊介绍： Aquaculture is an international journal for the exploration, improvement and management of all freshwater and marine food resources. It publishes novel and innovative research of world-wide interest on farming of aquatic organisms, which includes finfish, mollusks, crustaceans and aquatic plants for human consumption. Research on ornamentals is not a focus of the Journal. Aquaculture only publishes papers with a clear relevance to improving aquaculture practices or a potential application.