Kaabi M, Paz P, Alvarez M, Anel E, Boixo J C, Rouissi H, Herraez P, Anel L
Animal Reproduction and Obstetrics, University of León, 24071 León, Spain.
Theriogenology. 2003 Oct 15;60(7):1249-59. doi: 10.1016/s0093-691x(03)00139-0.
Post-mortem spermatozoa recovery is an important technique for obtaining germplasm reserves from genetically valuable animals or endangered species. However, there are many factors that influence the outcome of this technique. We have studied the effect of the interval between animal's death and sperm recovery (0, 24 or 48 h) on the quality and freezability of ram spermatozoa from cauda epididymidis. Storage temperature of epididymis (room temperature or 5 degrees C) was also analysed. Spermatozoa were diluted with Tes-Tris-Fructose solution supplemented with egg yolk (10%) and glycerol (4%), and frozen using a programmable biofreezer (-20 degrees C/min). Pre-freeze and post-thaw sperm samples showed viable spermatozoa up to 48 h after the animal's death, although their quality declined significantly as post-mortem storage time increased. Epididymis sperm stored at 5 degrees C showed better motility and a lower percentage of abnormal forms than epididymis stored at room temperature after 24 and 48 h. The fertilizing ability of cauda epididymis ram spermatozoa obtained at 0 and 24h after the animal's death is similar to that of ejaculated spermatozoa. Therefore, a good protocol for post-mortem semen collection in rams when epididymal spermatozoa cannot be collected immediately, is to preserve the epididymis at 5 degrees C and process the samples in the first 24h after the animal's death.
死后精子回收是从具有遗传价值的动物或濒危物种中获取种质资源的一项重要技术。然而，有许多因素会影响该技术的结果。我们研究了动物死亡与精子回收之间的间隔时间（0、24或48小时）对来自附睾尾部的公羊精子质量和冷冻保存能力的影响。还分析了附睾的储存温度（室温或5摄氏度）。精子用添加了蛋黄（10%）和甘油（4%）的Tes-三羟甲基氨基甲烷-果糖溶液稀释，并使用可编程生物冷冻机（每分钟降温20摄氏度）进行冷冻。尽管随着死后储存时间的增加精子质量显著下降，但在动物死亡后长达4小时的预冻和解冻后精子样本中仍显示有活精子。在24小时和48小时后，储存在5摄氏度的附睾精子比储存在室温下的附睾精子表现出更好的活力和更低的异常形态百分比。在动物死亡后0小时和24小时获得的附睾尾部公羊精子的受精能力与射出精子的受精能力相似。因此，当不能立即采集附睾精子时，公羊死后精液采集的一个良好方案是将附睾保存在5摄氏度，并在动物死亡后的头24小时内处理样本。
