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恢复方法和微生物污染对盘羊(Capra pyrenaica)附睾精子冷冻解冻反应的影响。

Influence of recovery method and microbial contamination on the response to freezing-thawing in ibex (Capra pyrenaica) epididymal spermatozoa.

机构信息

Departamento de Reproducción Animal, Avda. Puerta de Hierro Km 5.9, INIA, 28040 Madrid, Spain.

出版信息

Cryobiology. 2009 Dec;59(3):357-62. doi: 10.1016/j.cryobiol.2009.09.012. Epub 2009 Sep 26.

Abstract

The method of sperm recovery may influence the initial quality of sperm samples and their response to freezing-thawing. The aim of the present work was to compare two methods for collecting epididymal spermatozoa in order to improve the quality of recovered sperm and reduce possible contamination. Testes were obtained from 23 legally hunted, adult ibex males. The sperm mass of the right epididymis was collected by small longitudinal and transverse cuts made in the cauda epididymidis. The sperm mass of the left epididymis was collected by retrograde flushing from the vas deferens to the cauda epididymidis (using a cannula), employing a Tris, citric acid, glucose, egg yolk-based medium. The flushing method recovered more spermatozoa (P<0.001) than the cutting method. After freezing-thawing, greater acrosomes damage (P<0.001) and more morphological abnormalities (P<0.05) were seen among the sperm cells recovered by the cutting method than among those obtained by retrograde flushing. The method of sperm recovery did not, however, influence the microbial contamination rate. In frozen-thawed samples that were microbially contaminated, motility was significantly reduced (P<0.05) and membrane integrity tended to be poorer (P=0.06). In conclusion, retrograde flushing is recommended for ibex sperm collection since it would appear that microbial contamination is no more of a problem than that encountered with the cutting method, while a larger number of sperm cells more resistant to freezing-thawing can be obtained.

摘要

精子回收方法可能会影响精子样本的初始质量及其对冷冻-解冻的反应。本研究的目的是比较两种收集附睾精子的方法,以提高回收精子的质量并减少可能的污染。从 23 只合法猎捕的成年雄性野山羊睾丸中获得精子。通过在附睾尾部进行小的纵向和横向切口收集右侧附睾的精子质量,通过从输精管逆行冲洗到附睾尾部(使用套管)收集左侧附睾的精子质量,采用 Tris、柠檬酸、葡萄糖、蛋黄基培养基。冲洗法比切割法回收更多的精子(P<0.001)。冷冻-解冻后,切割法回收的精子中顶体损伤(P<0.001)和形态异常(P<0.05)比逆行冲洗法更多。然而,精子回收方法并未影响微生物污染率。在微生物污染的冷冻-解冻样本中,运动能力显著降低(P<0.05),膜完整性趋于较差(P=0.06)。总之,推荐使用逆行冲洗法收集野山羊精子,因为与切割法相比,微生物污染似乎不是问题,而且可以获得更多对冷冻-解冻更具抵抗力的精子。

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