Bielas Wiesław, Niżański Wojciech, Partyka Agnieszka, Rząsa Anna, Mordak Ryszard
Department of Reproduction and Clinic of Farm Animals, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, pl. Grunwaldzki 49, 50-366, Wrocław, Poland.
Department of Immunology, Pathophysiology and Veterinary Prevention Medicine, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, ul. C. K. Norwida 31, 50-375, Wrocław, Poland.
Acta Vet Scand. 2017 Sep 11;59(1):58. doi: 10.1186/s13028-017-0325-9.
There is some controversy about the extent of changes in different sperm cell features in stored boar semen, especially regarding the potential role of the DNA fragmentation assay for assessment of sperm fertilizing ability. The aim of this study was to assess the effect of time of storage and the dynamic changes in sperm cell characteristics in normospermic boar semen stored in long-term extender, in order to determine the susceptibility to damage of particular structures of spermatozoa during cooling and storage at 17 °C for 240 h post collection. The study included five ejaculates from each of seven boars of the Polish Large White breed (n = 35 ejaculates). The sperm characteristics were assessed using a flow cytometer and a computer assisted sperm analyzer on samples at 0, 48, 96, 168 and 240 h post collection.
The sperm chromatin structure assay (SCSA) showed a significant abrupt increase (P < 0.01) in the DNA fragmentation index (%DFI) after 48 h of semen storage with only subtle changes thereafter, not exceeding 5% on average after 240 h of storage. The use of a combination of SYBR-14/PI stains did not reveal any significant changes in the percentage of live sperm cells up to 168 h of semen storage. A significant (P < 0.01) decrease in the percentage of live spermatozoa with intact acrosomes was observed after prolonged semen storage (168 h). A significant and progressive decrease in sperm motility was recorded during the whole period of semen storage.
Storage of boar semen extended in long-term diluent at 17 °C for 48 h initially induced a decrease in the integrity of sperm DNA. This suggests that the structure of boar sperm DNA is susceptible to damage, especially during semen extension and at the beginning of sperm storage. These findings support the opinion that the SCSA test has only a low potential for routine assessment of boar semen preserved in the liquid state and for assessment of sperm quality changes during 10 days of semen preservation. Remarkably, the integrity of acrosomes and plasma membranes remained nearly unchanged for 7 days.
关于储存公猪精液中不同精子细胞特征的变化程度存在一些争议，特别是关于DNA碎片化检测在评估精子受精能力方面的潜在作用。本研究的目的是评估储存时间对公猪精液中精子细胞特征动态变化的影响，这些精液储存在长期稀释液中，以确定采集后在17°C下冷却和储存240小时期间精子特定结构对损伤的敏感性。该研究包括来自波兰大白猪品种的七头公猪中每头的五份射精样本（n = 35份射精样本）。在采集后0、48、96、168和240小时对样本使用流式细胞仪和计算机辅助精子分析仪评估精子特征。
精子染色质结构分析（SCSA）显示，精液储存48小时后，DNA碎片化指数（%DFI）显著急剧增加（P < ），此后仅有细微变化，储存240小时后平均不超过5%。使用SYBR - 14/PI染色组合在精液储存长达168小时时未发现活精子细胞百分比有任何显著变化。精液长时间储存（168小时）后，观察到顶体完整的活精子百分比显著（P < ）下降。在精液储存的整个期间记录到精子活力显著且逐渐下降。
公猪精液在17°C下用长期稀释液储存48小时最初会导致精子DNA完整性下降。这表明公猪精子DNA结构易受损伤，特别是在精液稀释和精子储存开始时。这些发现支持这样的观点，即SCSA检测在常规评估液态保存的公猪精液以及评估精液保存10天期间精子质量变化方面潜力较低。值得注意的是，顶体和质膜的完整性在7天内几乎保持不变。
