鸡细管冻精去甘油技术研究

Study on the Glycerol Remove Methods from Chicken Straw Frozen Semen

甘油细管冻精是鸡精液冷冻最有效的方案之一,但即使是迹量甘油也会影响受精率。本试验采用11%甘油浓度细管冷冻技术,自制便携式程序冷冻装置,在鸡冻精解冻后用清洗液分步稀释清洗、离心、重悬的方法去甘油,按解冻后精液与清洗液体积设置4种比例清洗(不清洗、1∶1、1∶2与1∶4),检测活力,并进行深阴部人工输精,捡蛋孵化并统计受精率与出雏率。结果表明:①4个组精子活力依次为1∶1组(0.4±0.05)>1∶2组(0.36±0.05)>不清洗组(0.32±0.08)>1∶4组(0.27±0.04),前3组均没有显著差异,只有1∶4组显著低于1∶1组(P<0.05);②4组受精率与出雏率依次为1∶2组(61.01%±13.35%、45.9%±10.74%)>1∶1组(52.37%±10.01%、45.9%±10.74%)>1∶4组(41.48%±11.17%、33.35%±10.41%)>未清洗组(6.29%±5.21%和5.03%±4.48%);③1∶2组显著高于1∶1组(P<0.05),极显著高于1∶4组(P<0.01)与未清洗组;④未清洗直接离心组极显著低于其它各组(P<0.01)。因此,采用分步稀释离心去甘油,解冻精液和稀释液比例为1∶2时,在5℃以600 g离心10 min,再人工输精,能够获得较好的受精率和出雏率。

It was one of the most effective methods to freeze rooster semen using glycerol and straw（SGS）.However,the fertility would be reduced by little amount of glycerol inside the semen.The SGS was made using the self-made portable freezing device.The glycerol（11%） was removed by stepwise dilution,centrifugating and resuspending after semen thawed.The four diluted ratios were made to remove the glycerol from the thawed semen,such as directly centrifugated,washed at the ratio of 1∶1,1∶2 and 1∶4（v/v） respectively.The sperm motility and intravaginal artificial insemination was made.It was indicated that：①The sperm motility order of the 4 groups was group 1∶1（0.4±0.05）〉group 1∶2（0.36±0.05）〉non-washed group（0.32±0.08）〉group 1∶4（0.27±0.04）,and there was no significant difference among the first 3 groups,only the group 1∶4 was significantly lower than that of the group 1∶1（P〈0.05）.②The fertility and hatchability（FH）order of the 4 groups was group 1∶2（61.01%±13.35%,45.9%±10.74%）〉group 1∶1（52.37%±10.01%,45.9%±10.74%）〉group 1∶4（41.48%±11.17%,33.35%±10.41%）〉non-washed group（6.29% ±5.21%and 5.03%±4.48%）.③The FH of group 1∶2 was significantly higher than that of group 1∶1（P〈0.05）,and it was extremely higher than that of group 1∶4（P〈0.01）and non-washed group.④The FH of non-washed group was extremely lower than that of any other groups（P〈0.01）.In conclusion,it could get good fertility and hatchability if the thawed SGS was washed at a ratio of 1∶2,then centrifuged for 10 min with a speed of 600 gat 5 ℃ and artificial inseminated.