Kasimanickam Ramanathan, Kasimanickam Vanmathy, Pelzer Kevin D, Dascanio John J
Department of Large Animal Clinical Sciences, Duckpond Drive, Phase III, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.
Anim Reprod Sci. 2007 Sep;101(1-2):60-73. doi: 10.1016/j.anireprosci.2006.09.001. Epub 2006 Oct 2.
The objectives of this study were (1) to determine the changes in structural, functional and motility parameters of ram-lamb semen stored at two different concentrations at 4 degrees C for 8 days in egg-yolk based extender and (2) to determine the effect of breed of ram-lambs on the changes in structural, functional and motility parameters of ram-lamb semen from different breeds stored at two different concentrations at 4 degrees C for 8 days in egg-yolk based extender. Two different concentrations suitable for laparoscopic and cervical insemination were employed in this experiment. A total of 14 ram-lambs (Polled Dorset-5, Suffolk-5, Katahdin-4) with satisfactory breeding potential were selected. Semen samples were collected by electro-ejaculation. Semen samples were extended to 50 and 200 million sperm per ml with a commercial egg yolk based extender (Triladyl, Minitube of America, Verona, WI, USA) at room temperature and were stored at 4 degrees C. The sperm DNA fragmentation index (DFI), percentages of high mitochondrial membrane potential (hMMP) and plasma membrane integrity (PMI) were assessed using flow cytometry as part of structural and functional parameters on Days 0, 1, 4, 6, and 8. A computer assisted sperm analyser (HTM-IVOS, Version 10.8, Hamilton Thorne Research, Beverly, MA, USA) was used to assess the sperm motility parameters on Days 0, 1, 4, 6, and 8. PROC MIXED procedure was used to determine the effect of days of storage, concentration and breed. The concentration and days of storage significantly affected the sperm structural, functional and motility parameters (P<0.0001). Significant concentration x days of storage interaction was found for all structural and functional parameters. There was a significant concentration x days of storage interaction for average path velocity, curvilinear velocity, straightness and linearity. Overall changes in the sperm structural, functional and sperm motility parameters over the storage period were less dramatic in the 200 x 10(6) ml(-1) concentration when compared to 50 x 10(6) ml(-1) concentration. The hMMP and total progressive motility were influenced by breed. In conclusion, the quality of structural, functional and motility parameters declined as days of storage were increased and the magnitude of changes in the parameters was less dramatic at the higher concentration.
（1）确定基于蛋黄的稀释液中，4℃下以两种不同浓度保存8天的公羊羔精液的结构、功能和活力参数的变化；（2）确定公羊羔品种对4℃下以两种不同浓度在基于蛋黄的稀释液中保存8天的不同品种公羊羔精液的结构、功能和活力参数变化的影响。本实验采用了两种适合腹腔镜授精和宫颈授精的不同浓度。总共挑选了14只具有良好繁殖潜力的公羊羔（无角陶赛特羊5只、萨福克羊5只、卡他丁羊4只）。通过电刺激采精收集精液样本。精液样本在室温下用市售的基于蛋黄的稀释液（Triladyl，美国Minitube公司，威斯康星州维罗纳）稀释至每毫升5000万和2亿精子，并在4℃下保存。在第0、1、4、6和第8天，使用流式细胞仪评估精子DNA碎片化指数（DFI）、高线粒体膜电位（hMMP）百分比和质膜完整性（PMI），作为结构和功能参数的一部分。使用计算机辅助精子分析仪（HTM-IVOS，版本10.8，美国马萨诸塞州贝弗利市汉密尔顿·桑恩研究公司）在第0、1、4、6和第8天评估精子活力参数。使用PROC MIXED程序确定保存天数、浓度和品种的影响。浓度和保存天数显著影响精子的结构、功能和活力参数（P<0.0001）。在所有结构和功能参数上均发现了显著的浓度×保存天数交互作用。在平均路径速度、曲线速度、直线性和线性方面存在显著的浓度×保存天数交互作用。与50×10⁶/ml⁻¹浓度相比，在200×10⁶/ml⁻¹浓度下，储存期间精子结构、功能和精子活力参数的总体变化较小。hMMP和总渐进性活力受品种影响。总之，随着保存天数的增加，结构、功能和活力参数的质量下降，且在较高浓度下参数变化的幅度较小。
