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影响公猪精子冷冻存活的因素。

Factors influencing boar sperm cryosurvival.

作者信息

Roca J, Hernández M, Carvajal G, Vázquez J M, Martínez E A

机构信息

Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, Campus de Espinardo, University of Murcia, E-30071 Murcia, Spain.

出版信息

J Anim Sci. 2006 Oct;84(10):2692-9. doi: 10.2527/jas.2006-094.

Abstract

Optimal sperm cryopreservation is a prerequisite for the sustainable commercial application of frozen-thawed boar semen for AI. Three experiments were performed to identify factors influencing variability of postthaw sperm survival among 464 boar ejaculates. Sperm-rich ejaculate fractions were cryopre-served using a standard freezing-thawing procedure for 0.5-mL plastic straws and computer-controlled freezing equipment. Postthaw sperm motility (assessed with a computer-assisted semen analysis system) and viability (simultaneously probed by flow cytometry analysis after triple-fluorescent stain), evaluated 30 and 150 min postthaw, were used to estimate the success of cryopreservation. In the first experiment, 168 unselected ejaculates (1 ejaculate/boar), from boars of 6 breeds with a wide age range (8 to 48 mo), were cryopreserved over a 12-mo period to evaluate the predictive value of boar (breed and age), semen collection, transport variables (season of ejaculate collection, interval between collections, and ejaculate temperature exposure), initial semen traits, and sperm quality before freezing on sperm survival after freezing-thawing. In Exp. 2, 4 ejaculates from each of 29 boars, preselected according to their initial semen traits and sperm quality before freezing, were collected and frozen over a 6-mo period to evaluate the influence of interboar and intraboar ejaculate variability in the survival of sperm after cryopreservation. In Exp. 3, 12 ejaculates preselected as for Exp. 2, from each of 15 boars with known good sperm cryosurvival, were collected and frozen over a 12-mo period to estimate the sustainability of sperm cryosurvival between ejaculates over time. Boar and semen collection and transport variables were not predictive of sperm cryosurvival among ejaculates. Initial semen traits and sperm quality variables observed before freezing explained 23.2 and 10.9%, respectively, of the variation in postthaw sperm motility and viability. However, more that 70% of total variance observed in postthaw sperm quality variables among ejaculates was explained by boar. This indicates that boar is the most important (P < 0.001) factor explaining the variability among ejaculates in sperm cryosurvival, with most (14 of the 15 boars in Exp. 3) showing consistent (P > 0.05) sperm cryosurvival over time.

摘要

优化猪精液冷冻保存是冷冻解冻后的公猪精液用于人工授精可持续商业应用的前提条件。进行了三项试验,以确定影响464份公猪射精样本解冻后精子存活变异性的因素。富含精子的射精部分采用标准冷冻解冻程序,在0.5毫升塑料细管中冷冻,并使用计算机控制的冷冻设备。解冻后30分钟和150分钟时,用计算机辅助精液分析系统评估解冻后精子活力,并用三荧光染色后通过流式细胞术分析同时检测精子活力,以此评估冷冻保存的成功率。在第一个试验中,在12个月期间对168份未筛选的射精样本(每头公猪1份射精样本)进行冷冻保存,这些样本来自6个品种、年龄范围广泛(8至48月龄)的公猪,以评估公猪(品种和年龄)、精液采集、运输变量(射精采集季节、采集间隔和射精温度暴露)、冷冻前初始精液特征和精子质量对冷冻解冻后精子存活的预测价值。在试验2中,根据冷冻前初始精液特征和精子质量预先选择了29头公猪,在6个月期间采集每头公猪的4份射精样本并进行冷冻,以评估公猪间和公猪内射精样本变异性对冷冻保存后精子存活的影响。在试验3中,在12个月期间采集预先选择的如试验2中的15头已知精子冷冻存活率良好的公猪的每份12份射精样本并进行冷冻,以估计射精样本间精子冷冻存活随时间的可持续性。公猪以及精液采集和运输变量不能预测射精样本间的精子冷冻存活情况。冷冻前观察到的初始精液特征和精子质量变量分别解释了解冻后精子活力和存活率变异的23.2%和10.9%。然而,射精样本间解冻后精子质量变量中观察到的总变异的70%以上可由公猪来解释。这表明公猪是解释射精样本间精子冷冻存活变异性的最重要因素(P<0.001),大多数公猪(试验3中的15头公猪中有14头)随时间显示出一致的(P>0.05)精子冷冻存活率。

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