Abstract

Aim of this study was to compare different combinations of penetrating intracellular CPAs, i.e., glycerol (G), ethylene glycol (EG), propylene glycol (PG), dimethyl formamide (DM), and methyl acetamide (MA) and extracellular [egg yolk (EY), egg yolk plasma (EYP), low-density lipoproteins (LDL), and coconut water (CW)] in Tris-citric acid-fructose buffer (T) for Labrador dog semen cryopreservation. The study was conducted in two parts, first trial was conducted to assess optimum glycerol concentration (5–7%) in TEY and equilibration time (ET, 2–4 hrs) for Labrador dog semen cryopreservation. Secondly, compatibility of 15% TEY, 15% TEYP, 13% TLDL, and 25% TCW with G, DMF, MA, D + M, EG, and PG was evaluated for in vitro sperm function tests. Decline in sperm attributes, i.e., motility, viability, plasma membrane integrity (PMI), and acrosome integrity (AI)) was significantly (p < 0.05) less in 7% TEY-G and 4 h compared to other concentrations and ET at post-thaw. There was significantly (p < 0.05) less decline in sperm attributes in TEY-G, TEYP-G, TLDL-G, TLDL-D, TLDL-EG, and TCW-D extenders compared to other combinations at post-thaw. However, these parameters were significantly (p < 0.05) high in TEY-G and TEYP-G compared to TEYP-D, TLDL-G, TLDL-D, TLDL-EG, and TCW-D extenders at post-thaw. However, decline in motility, viability, PMI, and AI was identical in these seven extenders. This study concluded that glycerol at a concentration of 7% in TEY and 4 h ET were optimum for successful cryopreservation and besides TEY-G, other combinations of protectants may be an alternative for canine semen cryopreservation.

摘要

本研究的目的是比较穿透细胞内 CPA 的不同组合，即甘油 (G)、乙二醇 (EG)、丙二醇 (PG)、二甲基甲酰胺 (DM) 和甲基乙酰胺 (MA) 以及细胞外 [蛋黄(EY)、蛋黄血浆 (EYP)、低密度脂蛋白 (LDL) 和椰子水 (CW)] 在三柠檬酸 - 果糖缓冲液 (T) 中用于拉布拉多犬的精液冷冻保存。该研究分两部分进行，第一个试验是评估 TEY 中的最佳甘油浓度（5-7%）和拉布拉多犬精液冷冻保存的平衡时间（ET，2-4 小时）。其次，评估了 15% TEY、15% TEYP、13% TLDL 和 25% TCW 与 G、DMF、MA、D + M、EG 和 PG 的体外相容性精子功能测试。 与其他浓度和 ET 相比，7% TEY-G 和 4 小时后精子属性，即运动性、活力、质膜完整性 (PMI) 和顶体完整性 (AI)) 的下降显着（p < 0.05） -解冻。 与解冻后的其他组合相比，TEY-G、TEYP-G、TLDL-G、TLDL-D、TLDL-EG 和 TCW-D 扩展剂的精子属性下降显着（p < 0.05）。然而，这些参数显着（p < 0.05) TEY-G 和 TEYP-G 在解冻后与 TEYP-D、TLDL-G、TLDL-D、TLDL-EG 和 TCW-D 扩展剂相比较高。然而，这七种扩展剂的运动性、活力、PMI 和 AI 的下降是相同的。该研究得出结论，TEY 中浓度为 7% 的甘油和 4 小时 ET 是成功冷冻保存的最佳选择，除 TEY-G 外，其他保护剂组合可能是犬精液冷冻保存的替代方案。