Pardede Berlin Pandapotan, Agil Muhammad, Yudi Yudi, Supriatna Iman
Reproductive Biology Study Program, Faculty of Veterinary Medicine, IPB University, Bogor, Indonesia.
Department of Veterinary Clinic, Reproduction, and Pathology, Division of Reproduction and Obstetrics, Faculty of Veterinary Medicine, IPB University, Bogor, Indonesia.
Vet World. 2020 Dec;13(12):2649-2657. doi: 10.14202/vetworld.2020.2649-2657. Epub 2020 Dec 14.
Various factors can reduce the quality of semen used for artificial insemination and have an impact on fertility decline, such as poor handling during frozen semen distribution. This study was aimed at assessing the quality of frozen-thawed semen after distribution in the field and its importance in maintaining fertility.
The Brahman Cross (BX) breeding program of PT Lembu Jantan Perkasa, Indonesia, was used. This program was preferred due to its adherence to guidelines that limit the effects of extraneous factors that may affect semen quality. Frozen-thawed semen samples from eight bulls with the same production code were analyzed and compared between the production site (artificial insemination [AI] center) and the field (BX breeding program). Total and progressive motility (PM) of sperm were determined using computer-assisted semen analysis. Plasma membrane integrity (PMI) was assessed using hypoosmotic swelling test, sperm viability using Eosin-Nigrosin staining, acrosome integrity using trypan blue-Giemsa staining, morphological abnormalities using William staining, and DNA fragmentation using toluidine blue staining. The fertility rate was determined using the conception rate (%) derived from AI data based on 502 AI services and 478 cows in the BX breeding program. A t-test was used to compare the quality of frozen-thawed semen before and after distribution. The relationship between the qualities of frozen semen after distribution in the field with fertility was analyzed using Pearson correlation.
There was no significant difference (p>0.05) in the quality of frozen-thawed semen (sperm motility, PMI, viability, acrosome integrity, abnormalities, and DNA fragmentation) between the production site (AI center) and after distribution in the field (BX breeding program). The semen met the minimum standards for AI programs. Total motility (r=0.986), PM (r=0.961), sperm viability (r=0.971), PMI (r=0.986), and acrosome integrity (r=0.992) were all positively correlated (p<0.05) with fertility rate; while sperm abnormalities (r=-0.996) and sperm DNA fragmentation (r=0.975) were negatively correlated (p<0.05) with fertility rate.
The study showed that to achieve the maximal and optimal fertility rate in bulls in an AI program, the overall quality of frozen-thawed semen in all aspects is critical. This can be achieved if the handling during distribution and storage, as well as the various factors that may affect the quality of semen in the field, can be controlled properly.
多种因素会降低用于人工授精的精液质量，并对生育能力下降产生影响，比如冷冻精液分发过程中的不当处理。本研究旨在评估野外分发后冻融精液的质量及其对维持生育能力的重要性。
采用印度尼西亚PT Lembu Jantan Perkasa公司的婆罗门杂交（BX）育种计划。选择该计划是因为其遵循相关准则，可限制可能影响精液质量的外部因素的影响。对来自8头具有相同生产编号的公牛的冻融精液样本进行分析，并在生产地点（人工授精[AI]中心）和野外（BX育种计划）之间进行比较。使用计算机辅助精液分析确定精子的总活力和渐进性活力（PM）。使用低渗肿胀试验评估质膜完整性（PMI），使用伊红 - 苯胺黑染色评估精子活力，使用台盼蓝 - 吉姆萨染色评估顶体完整性，使用威廉染色评估形态异常，使用甲苯胺蓝染色评估DNA片段化。根据BX育种计划中基于502次人工授精服务和478头母牛的人工授精数据确定受胎率（%）。使用t检验比较分发前后冻融精液的质量。使用Pearson相关性分析野外分发后冻融精液质量与生育能力之间的关系。
生产地点（AI中心）和野外分发后（BX育种计划）的冻融精液质量（精子活力、PMI、活力、顶体完整性、异常和DNA片段化）没有显著差异（p>0.05）。精液符合人工授精计划的最低标准。总活力（r = 0.986）、PM（r = 0.961）、精子活力（r = 0.971）、PMI（r = 0.986）和顶体完整性（r = 0.992）均与受胎率呈正相关（p<0.05）；而精子异常（r = -0.996）和精子DNA片段化（r = 0.975）与受胎率呈负相关（p<0.05）。
该研究表明，要在人工授精计划中使公牛达到最大和最佳受胎率，冻融精液在各个方面的整体质量至关重要。如果能妥善控制分发和储存过程中的处理以及可能影响野外精液质量的各种因素，就能实现这一点。
