Amorim E A M, Torres C A A, Graham J K, Amorim L S, Santos L V L
Department of Animal Science, Federal University of Vicosa, MG, Brazil.
Anim Reprod Sci. 2009 Apr;111(2-4):338-43. doi: 10.1016/j.anireprosci.2008.02.015. Epub 2008 Feb 26.
The hypoosmotic swelling test (HOST) has proved to be a good tool for evaluating the membrane integrity of spermatozoa of various domestic animals including cattle, horses, and swine. However, the best approach for using this technique in rabbit semen has not been tested. The present study aimed to establish the best hypoosmotic solution (HS) for testing membrane integrity in fresh rabbit semen. Sucrose solutions with the following osmolarities were used: 50, 60, 75, 100, 125 and 150mOsm/L. Semen samples (n=30) were collected from five mature White New Zealand rabbits (six collections per rabbit) at 72h intervals. After macroscopic evaluation, 10microL of semen was immediately added to 2mL of each solution and incubated for 1h at 37 degrees C. Sequentially, 20microL of semen diluted in HS were evaluated with oil immersion using a phase-contrast microscope. A total of 200 spermatozoa were counted in at least five different fields, and sperm tails were classified as non-coiled, coiled, and strongly coiled. The respective percentages of spermatozoa with coiled tails (coiled plus strongly coiled) in the six solutions listed above were 54.8, 65.2, 54.3, 53.9, 38.9 and 29.4%. Percentage of strongly coiled spermatozoa was: 40.2, 51.0, 43.2, 41.5, 32.7 and 26.9 for the six solutions, respectively. According to total and strong coiling 60mOsm/L was superior to others treatments (P<0.05). Results suggest that the 60mOsm/L solution would be most desirable for use in HOST in fresh rabbit spermatozoa.
低渗肿胀试验（HOST）已被证明是评估包括牛、马和猪在内的各种家畜精子膜完整性的良好工具。然而，在兔精液中使用该技术的最佳方法尚未得到测试。本研究旨在确定用于测试新鲜兔精液膜完整性的最佳低渗溶液（HS）。使用了具有以下渗透压的蔗糖溶液：50、60、75、100、125和150mOsm/L。每隔72小时从五只成年新西兰白兔（每只兔子采集六次）采集精液样本（n = 30）。经过宏观评估后，立即将10微升精液添加到每种溶液的2毫升中，并在37摄氏度下孵育1小时。随后，使用相差显微镜油镜评估在HS中稀释的20微升精液。在至少五个不同视野中总共计数200个精子，并将精子尾部分类为非卷曲、卷曲和强烈卷曲。上述六种溶液中具有卷曲尾巴（卷曲加强烈卷曲）的精子的各自百分比分别为54.8%、65.2%、54.3%、53.9%、38.9%和29.4%。六种溶液中强烈卷曲精子的百分比分别为：40.2%、51.0%、43.2%、41.5%、32.7%和26.9%。根据总卷曲和强烈卷曲情况，60mOsm/L优于其他处理（P<0.05）。结果表明，60mOsm/L溶液最适合用于新鲜兔精子的低渗肿胀试验。
