Santiago-Moreno J, Coloma M A, Dorado J, Pulido-Pastor A, Gómez-Guillamon F, Salas-Vega R, Gómez-Brunet A, López-Sebastián A
Departamento de Reproducción Animal, Avda. Puerta de Hierro Km 5.9, INIA, 28040 Madrid, Spain.
Theriogenology. 2009 May;71(8):1253-60. doi: 10.1016/j.theriogenology.2008.12.019. Epub 2009 Feb 23.
The effectiveness of electroejaculation for obtaining Spanish ibex sperm samples for freeze preserving outside the rutting season was evaluated-the aim being to optimise biological resources for the establishment of germplasm banks. The effect of different egg yolk concentrations (6% or 12%, v/v) in diluents of different buffer composition (Tris-citric acid buffer or Tes-Tris buffer) on frozen-thawed samples of the above also investigated. Experiments were undertaken with six ibex males in February-May, and involved four different semen samples from each animal with four combination of extender, respectively: Tes-Tris-glucose (TTG)-6% egg yolk, TTG-12% egg yolk, Tris-citric acid-glucose (TCG)-12% egg yolk, TCG-6% egg yolk. The results show that electroejaculation is a useful way of obtaining sperm samples from Spanish ibex outside the rutting season (i.e., at a time coinciding with plasma testosterone levels of <0.4ng/ml). According to the results of the eosin-nigrosin staining and the hypo-osmotic swelling test, the freezing-thawing process significantly reduced the viability and membrane integrity of the spermatozoa extended with TTG-6% egg yolk, TTG-12% egg yolk, and TCG-12% egg yolk, but did not affect these variables in spermatozoa extended with TCG-6% egg yolk. Therefore, the use of Tris-citric acid-based extenders containing low concentrations of egg yolk is recommended for cryopreserving Spanish ibex spermatozoa obtained by electroejaculation outside the rutting season.
评估了电刺激采精法在非发情季节获取西班牙羱羊精子样本用于冷冻保存的有效性,目的是优化生物资源以建立种质库。还研究了不同缓冲液组成(Tris - 柠檬酸缓冲液或Tes - Tris缓冲液)的稀释液中不同蛋黄浓度(6%或12%,v/v)对上述冷冻解冻样本的影响。在2月至5月期间对6只雄性羱羊进行了实验,每只动物分别有4种不同的精液样本,分别与4种稀释剂组合:Tes - Tris - 葡萄糖(TTG) - 6%蛋黄、TTG - 12%蛋黄、Tris - 柠檬酸 - 葡萄糖(TCG) - 12%蛋黄、TCG - 6%蛋黄。结果表明,电刺激采精法是在非发情季节(即血浆睾酮水平<0.4ng/ml时)从西班牙羱羊获取精子样本的有效方法。根据伊红 - 黑色素染色和低渗肿胀试验结果,冻融过程显著降低了用TTG - 6%蛋黄、TTG - 12%蛋黄和TCG - 12%蛋黄稀释的精子的活力和膜完整性,但对用TCG - 6%蛋黄稀释的精子的这些变量没有影响。因此,建议使用含低浓度蛋黄的基于Tris - 柠檬酸的稀释剂来冷冻保存通过电刺激采精法在非发情季节获取的西班牙羱羊精子。