Williamson R G, Etches R J, Reinhart B S, MacPherson J W
Reprod Nutr Dev (1980). 1981;21(6B):1033-42. doi: 10.1051/rnd:19810801.
Semen was collected from Single Comb White Leghorn roosters, diluted 1:4 with Beltsville Poultry Semen Extender at 35 degrees C and cooled at various rates to 5 degrees C. DMSO was added to the semen between 10 and 120 min after ejaculation at temperatures between 15 and 5 degrees C, respectively. Following the addition of DMSO, the semen was allowed to equilibrate for 2 h in a 5 degrees C environment. The semen was then frozen at 1 degree C per min from 5 degrees C to - 20 degrees C, transferred into liquid nitrogen vapour for 4 to 10 min and then immersed in liquid nitrogen for 4 to 60 days. The thawed semen was inseminated on two consecutive days and fertility was calculated during 5 and 7 days commencing on the second day after the last insemination. In general, fertility was unaffected by the rate of cooling and the temperature at which the DMSO was added. In one trial, however, the fertilizing capacity was significantly greater if the DMSO was added within 45 min when the temperature of the semen was 15 degrees C. Approximately 42 to 71 p. 100 of eggs laid by hens during the 5 day period after the second insemination were fertile regardless of the method of cooling or the temperature of the semen when the DMSO was added. The motility of the ejaculates after thawing and before insemination varied between 15 and 55 p. 100 and this measure of physiological quality was a poor indicator of fertilizing capacity. Embryonic mortality was unaffected by the cooling and freezing procedure and hatchability of fertile eggs was not significantly different for the hens inseminated with either fresh or frozen semen.
精液采自单冠白来航公鸡,在35摄氏度下用贝尔茨维尔家禽精液稀释剂按1:4稀释,并以不同速率冷却至5摄氏度。分别在射精后10至120分钟、精液温度在15至5摄氏度之间时向精液中添加二甲基亚砜(DMSO)。添加DMSO后,精液在5摄氏度环境中平衡2小时。然后精液从5摄氏度以每分钟1摄氏度的速度冷冻至零下20摄氏度,转移到液氮蒸气中4至10分钟,然后浸入液氮中4至60天。解冻后的精液连续两天进行输精,并在最后一次输精后第二天开始的第5天和第7天计算受精率。一般来说,冷却速率和添加DMSO的温度对受精率没有影响。然而,在一项试验中,如果在精液温度为15摄氏度时45分钟内添加DMSO,受精能力会显著提高。第二次输精后5天内母鸡所产鸡蛋中,约42%至71%受精,无论冷却方法或添加DMSO时精液的温度如何。解冻后输精前射精精液的活力在15%至55%之间变化,这种生理质量指标对受精能力的指示作用较差。胚胎死亡率不受冷却和冷冻程序的影响,用新鲜或冷冻精液输精的母鸡所产受精卵的孵化率没有显著差异。