Department of Veterinary Gynecology and Obstetrics, G. B. Pant University of Agriculture and Technology, Pantnagar, India.
Department of Livestock Production Management, and G. B. Pant University of Agriculture and Technology, Pantnagar, India.
Biopreserv Biobank. 2023 Aug;21(4):336-345. doi: 10.1089/bio.2022.0013. Epub 2022 Jul 19.
The present study was conducted to observe the effects of removal of seminal plasma of Pantja buck semen and supplementation of bovine seminal plasma (BSP) in the extender before cryopreservation. In a preliminary experiment, different levels of BSP were supplemented (1, 3, 5, 7, and 9% v/v) in egg yolk (7.5% egg yolk)-tris (EYT) extender and used for cryopreservation of Pantja buck semen. Results in terms of motility, viability, plasma membrane integrity, acrosome integrity, and lipid peroxidation showed that 5% BSP was suitable for maintaining Pantja buck semen quality during cryopreservation. In the final experiment, pooled semen from four Pantja bucks was split into three aliquots (I, II, and III). Aliquot I was directly diluted in EYT extender and grouped as the control (C); aliquot II and III were washed separately with TALP solution and diluted as D1 (Washed semen with EYT extender) and D2 (Washed semen with EYT extender containing 5% BSP), respectively. Seminal attributes (sperm individual motility, viability, plasma membrane integrity, acrosome integrity, and total morphological abnormalities) were assessed at the postdilution, postequilibration, and post-thawing stages. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA) concentration, and glutathione peroxidase (GSH-Px) activity were measured at post-thaw. Washed semen significantly improved ( < 0.05) seminal parameters at post-thaw compared with unwashed semen (control). A significant difference ( < 0.05) was observed in seminal attributes between freezing stages and between dilution groups. Significantly higher ( < 0.05) post-thaw sperm motility, viability, plasma membrane integrity, acrosome integrity, and GSH-Px activity, and significantly lower ( < 0.05) MDA concentration and extracellular release of enzymes (ALT, AST) were observed in group D2 compared with control and D1. The results of the present study demonstrated that cryopreservation of washed Pantja buck semen diluted with 5% BSP-supplemented EYT extender can improve post-thaw semen quality.
本研究旨在观察去除 Pantja 公鹿精液中的精浆和在冷冻前添加牛精浆（BSP）对稀释液的影响。在初步实验中，在卵黄（7.5%卵黄）-三（EYT）稀释液中添加不同水平的 BSP（1%、3%、5%、7%和 9%v/v），并用于冷冻 Pantja 公鹿精液。在活力、存活率、质膜完整性、顶体完整性和脂质过氧化方面的结果表明，5%的 BSP 适合在冷冻过程中维持 Pantja 公鹿精液的质量。在最后的实验中，从四只 Pantja 公鹿中收集的精液分成三份（I、II 和 III）。I 份直接在 EYT 稀释液中稀释，并作为对照组（C）；II 份和 III 份分别用 TALP 溶液洗涤，然后分别在 EYT 稀释液（洗涤精液）和 EYT 稀释液中添加 5%BSP（洗涤精液）中稀释，D2。在稀释后、平衡后和解冻后阶段评估精液属性（精子个体活力、存活率、质膜完整性、顶体完整性和总形态异常）。解冻后测量丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、丙二醛（MDA）浓度和谷胱甘肽过氧化物酶（GSH-Px）活性。与未洗涤精液（对照组）相比，洗涤精液在解冻后显著改善了（ <0.05）精液参数。在不同的冷冻阶段和稀释组之间观察到精液属性存在显著差异（ <0.05）。与对照组和 D1 相比，D2 组解冻后精子活力、存活率、质膜完整性、顶体完整性和 GSH-Px 活性显著升高（ <0.05），MDA 浓度和细胞外酶释放（ALT、AST）显著降低（ <0.05）。本研究结果表明，用添加 5%BSP 的 EYT 稀释液洗涤 Pantja 公鹿精液并冷冻，可以提高解冻后精液的质量。
