Borg K, Colenbrander B, Fazeli A, Parlevliet J, Malmgren L
Department of Obstetrics and Gynaecology Swedish University of Agricultural Sciences Uppsala, Sweden.
Theriogenology. 1997 Sep;48(4):531-6. doi: 10.1016/s0093-691x(97)00269-0.
Different thawing methods are used for stallion semen, however, it is unclear which method is the optimal one. To determine if the thawing temperature has an effect on semen quality, we compared 2 thawing temperatures, 75 degrees C and 37 degrees C. The following parameters were used to measure sperm quality: sperm motility, sperm viability, plasma membrane integrity and sperm morphology. Twenty-three ejaculates from 10 Dutch Warmblood stallions were thawed either at 37 degrees C for 30 sec or at 75 degrees C for 7 sec. Sperm motility was evaluated by a Hamilton Thorn Motility Analyser. Plasma membrane integrity and sperm viability were evaluated by using a live/dead fluorescein stain containing a calcein AM probe and ethidium homodimer-1 probe. The eosinaniline blue staining method was used to evaluate the percentage of live and dead cells, as well as sperm morphology. There was no significant difference (P = 0.84) between sperm motility after thawing at 37 degrees C and 75 degrees C. There was also no significant difference (P = 0.053) between the percentage of live spermatozoa using the calcein AM/ethidium homodimer stain after thawing at 37 degrees C and 75 degrees C. There was, however, a significant difference (P = 0.032) between the percentage of live spermatozoa using the eosin-aniline blue stain after thawing at 37 degrees C compared with that at 75 degrees C. In conclusion, our laboratory results indicated that stud farms using frozen semen should thaw the straws at 37 degrees C instead of 75 degrees C. The lower temperature is easier to work with, as thawing at the higher temperature requires special equipment and has to be timed very carefully to avoid damage to the spermatozoa.
种公马精液采用不同的解冻方法，然而，尚不清楚哪种方法是最佳方法。为了确定解冻温度是否对精液质量有影响，我们比较了75摄氏度和37摄氏度这两种解冻温度。使用以下参数来衡量精子质量：精子活力、精子存活率、质膜完整性和精子形态。来自10匹荷兰温血种公马的23份射精样本分别在37摄氏度下解冻30秒或在75摄氏度下解冻7秒。精子活力通过汉密尔顿·桑恩活力分析仪进行评估。质膜完整性和精子存活率通过使用含有钙黄绿素乙酰甲酯探针和乙锭同二聚体-1探针的活/死荧光素染色来评估。采用伊红苯胺蓝染色法评估活细胞和死细胞的百分比以及精子形态。在37摄氏度和75摄氏度下解冻后的精子活力之间没有显著差异（P = 0.84）。在37摄氏度和75摄氏度下解冻后使用钙黄绿素乙酰甲酯/乙锭同二聚体染色的活精子百分比之间也没有显著差异（P = 0.053）。然而，在37摄氏度下解冻后与75摄氏度下解冻后使用伊红-苯胺蓝染色的活精子百分比之间存在显著差异（P = 0.032）。总之，我们的实验室结果表明，使用冷冻精液的种马场应在37摄氏度而不是75摄氏度下解冻细管。较低的温度更易于操作，因为在较高温度下解冻需要特殊设备，并且必须非常精确地计时以避免对精子造成损害。
