Sadeghi Sara, Del Gallego Raquel, García-Colomer Balma, Gómez Ernesto A, Yániz Jesús L, Gosálvez Jaime, López-Fernández Carmen, Silvestre Miguel A
Departamento de Biología Celular, Biología Funcional y Antropología Física, Universitat de València, 46100 Valencia, Spain.
Centro de Investigación y Tecnología Animal, Instituto Valenciano de Investigaciones Agrarias, Pol. La Esperanza 100, 12400 Castellón, Spain.
Biology (Basel). 2020 Sep 19;9(9):300. doi: 10.3390/biology9090300.
The use of cooled semen is relatively common in goats. There are a number of advantages of cooled semen doses, including easier handling of artificial insemination (AI) doses, transport, more AI doses per ejaculate, and higher fertility rates in comparison with frozen AI doses. However, cooled semen has a short shelf life. The objective of this study was to examine the effect of temperature and sperm concentration on the in vitro sperm quality during liquid storage for 48 h, including sperm motility and kinetics, response to oxidation, mitochondrial membrane potential (MMP) and DNA fragmentation in goats. Three experiments were performed. In the first, the effects of liquid preservation of semen at different temperatures (5 °C or 17 °C), durations (0, 24 and 48 h) and sperm concentrations (250 × 10 sperm/mL (1:2 dilution rate), 166.7 × 10 sperm/mL (1:3 dilution rate) or 50 × 10 sperm/mL (1:10 dilution rate)) on sperm motility and kinetics were studied. In the second experiment, the effect of temperature, sperm washing and concentration on sperm motility and DNA fragmentation was studied. Finally, the effect of sperm concentration and duration of storage at 5 °C on sperm motility, response to oxidative stress and MMP was examined. We found that refrigerated liquid storage of goat sperm impaired sperm quality, such as motility, MMP and response to oxidation, as storage time increased; however, sperm DNA fragmentation index was not significantly affected. Liquid storage at 5 °C preserved higher total motility than at 17 °C. Moreover, we observed that the reduction of sperm concentration below 500 × 10 sperm/mL did not seem to improve the quality of spermatozoa conserved in milk-based extender in the conditions tested.
在山羊中，使用冷藏精液相对普遍。冷藏精液剂量有许多优点，包括人工授精（AI）剂量更易于处理、运输，每次射精可获得更多AI剂量，并且与冷冻AI剂量相比生育率更高。然而，冷藏精液的保质期较短。本研究的目的是检查温度和精子浓度对山羊精液在液体保存48小时期间的体外精子质量的影响，包括精子活力和动力学、氧化反应、线粒体膜电位（MMP）和DNA片段化。进行了三个实验。第一个实验研究了精液在不同温度（5℃或17℃）、保存时间（0、24和48小时）和精子浓度（250×10⁶精子/mL（1:2稀释率）、166.7×10⁶精子/mL（1:3稀释率）或50×10⁶精子/mL（1:10稀释率））下液体保存对精子活力和动力学的影响。在第二个实验中，研究了温度、精子洗涤和浓度对精子活力和DNA片段化的影响。最后，检查了精子浓度和在5℃下保存时间对精子活力、氧化应激反应和MMP的影响。我们发现，随着保存时间的增加，山羊精子的冷藏液体保存会损害精子质量，如活力、MMP和氧化反应；然而，精子DNA片段化指数没有受到显著影响。在5℃下液体保存比在17℃下保留了更高的总活力。此外，我们观察到，在测试条件下，将精子浓度降低至500×10⁶精子/mL以下似乎并未改善保存在基于牛奶的稀释剂中的精子质量。
