Department of Medicine and Animal Surgery, Faculty of Veterinary Science, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, Spain.
Anim Reprod Sci. 2012 May;132(1-2):66-73. doi: 10.1016/j.anireprosci.2012.04.003. Epub 2012 Apr 13.
The present study aimed to evaluate the ability of spermatozoa from individual boar ejaculates to withstand different semen-processing techniques. Eighteen sperm-rich ejaculate samples from six boars (three per boar) were diluted in Beltsville Thawing Solution and split into three aliquots. The aliquots were (1) further diluted to 3×10(7) sperm/mL and stored as a liquid at 17°C for 72 h, (2) frozen-thawed (FT) at 1×10(9) sperm/mL using standard 0.5-mL straw protocols, or (3) sex-sorted with subsequent liquid storage (at 17°C for 6 h) or FT (2×10(7) sperm/mL using a standard 0.25-mL straw protocol). The sperm quality was evaluated based on total sperm motility (the CASA system), viability (plasma membrane integrity assessed using flow cytometry and the LIVE/DEAD Sperm Viability Kit), lipid peroxidation (assessed via indirect measurement of the generation of malondialdehyde (MDA) using the BIOXYTECH MDA-586 Assay Kit) and DNA fragmentation (sperm chromatin dispersion assessed using the Sperm-Sus-Halomax(®) test). Data were normalized to the values assessed for the fresh (for liquid-stored and FT samples) or the sorted semen samples (for liquid stored and the FT sorted spermatozoa). All of the four sperm-processing techniques affected sperm quality (P<0.01), regardless of the semen donor, with reduced percentages of motile and viable sperm and increased MDA generation and percentages of sperm with fragmented DNA. Significant (P<0.05) inter-boar (effect of boars within each semen-processing technique) and intra-boar (effect of semen-processing techniques within each boar) differences were evident for all of the sperm quality parameters assessed, indicating differences in the ability of spermatozoa from individual boars to withstand the semen-processing techniques. These results are the first evidence that ejaculate spermatozoa from individual boars can respond in a boar-dependent manner to different semen-processing techniques.
本研究旨在评估来自单个公猪精液的精子在不同精液处理技术下的耐受能力。从六头公猪(每头三头)中采集了 18 份富含精子的精液样本,将其稀释在贝尔茨维尔解冻液中并分为三份。这些样本(1)进一步稀释至 3×10(7)个精子/mL,并以液体形式储存在 17°C 下 72 小时,(2)使用标准的 0.5-mL straw 方案以 1×10(9)个精子/mL 进行冷冻-解冻(FT),或(3)使用标准的 0.25-mL straw 方案以 2×10(7)个精子/mL 进行性别分选,随后进行液体储存(在 17°C 下储存 6 小时)或 FT。根据总精子活力(CASA 系统)、活力(使用流式细胞术和 LIVE/DEAD 精子活力试剂盒评估质膜完整性)、脂质过氧化(通过间接测量丙二醛(MDA)的生成来评估,使用 BIOXYTECH MDA-586 测定试剂盒)和 DNA 碎片化(使用 Sperm-Sus-Halomax(®)测试评估精子染色质分散)来评估精子质量。数据被归一化为新鲜精液(用于液体储存和 FT 样本)或分选精液样本(用于液体储存和 FT 分选精子)评估的值。所有四种精液处理技术都会影响精子质量(P<0.01),与精液供体无关,导致可运动和存活的精子比例降低,MDA 生成增加,DNA 碎片化的精子比例增加。所有评估的精子质量参数都显示出显著的(P<0.05)公猪间(每种精液处理技术内的公猪效应)和公猪内(每种公猪内的精液处理技术效应)差异,表明来自单个公猪的精子在耐受精液处理技术方面存在差异。这些结果首次证明,来自单个公猪的精液精子可以以公猪依赖的方式对不同的精液处理技术做出反应。