Pugliesi G, Fürst R, Carvalho G R
Department of Animal Reproduction, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, Pirassununga, SP, Brazil.
Andrologia. 2014;46(9):1055-62. doi: 10.1111/and.12205. Epub 2013 Dec 8.
The effects of freezing technique and thawing protocol on thawed semen viability and fertility were studied. Ejaculates from 5 stallions (n = 25) were frozen by conventional or a fast-freezing technique. Frozen semen was thawed by two thawing protocols (37 °C 30 s(-1) or 75 °C 7 s(-1) ). Thawed semen was evaluated by progressive motility, vigour, morphology and plasma membrane integrity. Mares (n = 25) were inseminated with 300 (n = 11) or 150 (n = 14) million spermatozoa. A greater (P < 0.05) vigour and progressively motile spermatozoa were detected, respectively, at thawing and after 20 min post-thawing in the fast-freezing technique than in the conventional one. Plasma membrane integrity was also greater (P < 0.05) in semen frozen with the fast-freezing technique. Semen viability was not affected by thawing protocol. Pregnancy rate using the fast-freezing technique was 76% (19/25), and did not differ (P > 0.05) between insemination doses. We concluded that the 150 million progressively motile spermatozoa per dose using a deep-horn insemination maximises the use of equine semen. The fast-freezing technique, as compared to the conventional one, efficiently preserves the viability and fertilising capacity of spermatozoa, indicating a new method to improve the fertility of frozen equine semen.
研究了冷冻技术和解冻方案对解冻后精液活力和受精能力的影响。采集了5匹种马的射精样本（n = 25），分别采用传统冷冻技术或快速冷冻技术进行冷冻。冷冻精液通过两种解冻方案进行解冻（37°C 30秒(-1)或75°C 7秒(-1)）。解冻后的精液通过精子的前向运动能力、活力、形态和质膜完整性进行评估。对25匹母马分别输入3亿（n = 11）或1.5亿（n = 14）个精子进行授精。与传统冷冻技术相比，快速冷冻技术在解冻时以及解冻后20分钟检测到的精子活力和前向运动精子数量分别更高（P < 0.05）。采用快速冷冻技术冷冻的精液质膜完整性也更高（P < 0.05）。精液活力不受解冻方案的影响。采用快速冷冻技术的妊娠率为76%（19/25），不同授精剂量之间无显著差异（P > 0.05）。我们得出结论，每剂量1.5亿个前向运动精子采用子宫角深部授精可最大限度地利用马精液。与传统冷冻技术相比，快速冷冻技术能有效保存精子的活力和受精能力，表明这是一种提高冷冻马精液受精能力的新方法。
