Mujitaba Malam Abulbashar, Kútvölgyi Gabriella, Radnai Szentpáli Judit, Debnár Viktória Johanna, Tokár Alexandra, Vass Nóra, Bodó Szilárd
Department of Animal Nutrition and Physiology, Faculty of Agriculture and Food Sciences and Environmental Management, University of Debrecen, Böszörményi Street 138, H-4032 Debrecen, Hungary.
Doctoral School of Animal Science, University of Debrecen, H-4032 Debrecen, Hungary.
Animals (Basel). 2024 Apr 20;14(8):1237. doi: 10.3390/ani14081237.
There are limited studies on the factors affecting the success of ram epididymal spermatozoa (REPS) cryopreservation. On this note, the current study assessed the influence of three commercial soy lecithin-based semen extenders, AndroMed (AND), BioXcell (BIO), and OviXcell (OVI), and two concentrations (400 × 10 vs. 200 × 10 spermatozoa/mL) on the pre-freeze and post-thaw quality of REPS. The REPS were retrieved from nine adult rams' testes and diluted with each of the three extenders to both concentrations. Straws were frozen manually. Standard motility (SMP) and kinematic parameters (KPs) were assessed via a CASA, while spermatozoa viability, morphology, and acrosomal integrity were assessed via the Kovács-Foote staining technique. The concentration did not significantly affect the pre-freeze and post-thaw SMP and KPs of REPS. BIO and OVI had significantly higher pre-freeze and post-thaw BCFs, post-thaw VAP, and the percentage of all intact heads than AND. In contrast, AND had a significantly lower percentage of REPS with tail defects than BIO and OVI. The 400 × 10 spermatozoa/mL concentration resulted in a significantly higher percentage of all intact heads than the 200 × 10 spermatozoa/mL concentration. Freezing significantly increased tail defects and decreased the percentage of REPS with distal cytoplasmic droplets. The cryopreservation of REPS at the 400 × 10 spermatozoa/mL concentration is recommended. All three extenders must be optimized to preserve the viability, membrane integrity, and better normal morphology of REPS; the reason for increased tail abnormality after the freezing/thawing process needs to be studied.
关于影响附睾精子（REPS）冷冻保存成功的因素，相关研究有限。基于此，本研究评估了三种商业化的大豆卵磷脂基精液稀释剂，即AndroMed（AND）、BioXcell（BIO）和OviXcell（OVI），以及两种浓度（400×10与200×10精子/mL）对REPS冻前和冻后质量的影响。从九只成年公羊的睾丸中采集REPS，并用三种稀释剂分别将其稀释至两种浓度。细管采用手动冷冻。通过计算机辅助精子分析（CASA）评估标准活力（SMP）和运动学参数（KPs），同时通过科瓦奇-富特染色技术评估精子活力、形态和顶体完整性。浓度对REPS的冻前和冻后SMP及KPs没有显著影响。BIO和OVI的冻前和冻后曲线速度（BCFs）、冻后平均路径速度（VAP）以及所有完整头部的百分比均显著高于AND。相比之下，AND的REPS尾部缺陷百分比显著低于BIO和OVI。400×10精子/mL浓度组的所有完整头部百分比显著高于200×10精子/mL浓度组。冷冻显著增加了尾部缺陷，并降低了带有远端细胞质滴的REPS百分比。建议以400×10精子/mL浓度冷冻保存REPS。所有三种稀释剂都必须进行优化，以保持REPS的活力、膜完整性和更好的正常形态；冷冻/解冻过程后尾部异常增加的原因有待研究。
