Kulíková Barbora, Oravcová Marta, Baláži Andrej, Supuka Peter, Chrenek Peter
Research Institute for Animal Production,National Agricultural and Food Centre,Nitra,Hlohovecka 2,951 41 Lužianky,Slovak Republic.
University of Veterinary Medicine and Pharmacy in Košice,Komenského 73,041 81 Košice,Slovak Republic.
Zygote. 2017 Oct;25(5):592-600. doi: 10.1017/S0967199417000454. Epub 2017 Aug 24.
In this study, fresh and frozen-thawed semen of Nitra and Zobor rabbit breeds were evaluated for potential inter-breed or inter-male differences in sperm quality traits. Individual male semen from four rabbits of each breed were diluted (v:v; 1:1) in a freezing medium composed of a commercial diluent, 16% of dimethyl sulphoxide (DMSO), 4% of Ficoll 70 and 2% of sucrose and frozen in liquid nitrogen vapours before being plunged into liquid nitrogen. Different motility traits, viability and plasma membrane integrity of fresh and frozen-thawed semen were evaluated in vitro using computer-assisted sperm analysis and flow cytometry. To evaluate the sperm fertilization ability, artificial insemination of fresh and frozen-thawed sperm was performed. Our results showed the effect of breed (P ≤ 0.05) on frozen-thawed sperm viability and plasma membrane integrity. Moreover, individual variability in semen quality among the rabbits was revealed (0.31 to 0.71 among quality traits). Our results thereby confirmed that the cryopreservation procedure could not ensure comparable sperm post-thaw survival for different breeds or males. Nevertheless, correlations between numbers of fresh total motile and progressively moving sperm and several quality parameters measured post thawing were revealed. Therefore, we suggest that the objective assessment of fresh rabbit sperm motility may be an effective indicator of frozen-thawed semen quality. Consequently, regular semen assessment is required in order to preserve good-quality insemination doses from native breeds.
在本研究中，对尼特拉兔和佐博尔兔品种的新鲜精液及冻融精液进行了评估，以探究品种间或个体间精子质量特征的潜在差异。将每个品种四只兔子的精液个体样本，用由商业稀释剂、16%二甲亚砜（DMSO）、4%聚蔗糖70和2%蔗糖组成的冷冻培养基按1:1（体积比）稀释，并在液氮蒸汽中冷冻，然后投入液氮。使用计算机辅助精子分析和流式细胞术在体外评估新鲜精液及冻融精液的不同活力特征、存活率和质膜完整性。为评估精子的受精能力，对新鲜精液及冻融精液进行了人工授精。我们的结果显示品种对冻融精子的存活率和质膜完整性有影响（P≤0.05）。此外，还揭示了兔子精液质量的个体差异（质量特征之间的差异为0.31至0.71）。我们的结果因此证实，冷冻保存程序无法确保不同品种或个体的精子解冻后有可比的存活率。然而，新鲜精液中总活动精子数和渐进性活动精子数与解冻后测量的几个质量参数之间存在相关性。因此，我们建议对新鲜兔精子活力进行客观评估可能是冻融精液质量的有效指标。因此，为了保存来自本地品种的高质量授精剂量，需要定期进行精液评估。
