不同脱甘油方法对鸡冷冻精液人工授精效果的影响

doi:10.16431/j.cnki.1671-7236.2025.08.019

摘要/Abstract

摘要： 【目的】探究不同甘油脱除方法对冻融后鸡精液人工授精效果的影响，以简化鸡精液的脱甘油程序。【方法】通过腹背联合按摩法采集文昌公鸡精液，冷冻保存后解冻，并用稀释液按1∶2、1∶4、1∶6、1∶8(V/V)的比例稀释去除甘油，通过检测脱甘油后的精子活力确定最佳稀释比。按最适比例分别采用一步、两步和六步稀释法进行脱甘油处理，通过检测精子活力、精子回收率、精清和精子中的甘油含量评价不同方法的脱甘油效果；将3种不同方法去除甘油的精液进行人工授精试验，检测受精率和孵化率。【结果】在不同稀释比例中，按1∶4六步稀释法去甘油，脱甘油后及脱甘油后30 min的精子活力相较于1∶2、1∶6、1∶8六步稀释法维持较好，但差异不显著(P＞0.05)。在总稀释比为1∶4的不同脱甘油方法中，六步稀释法脱甘油后精子活力最高，达到了68.34%，但3种方法间差异并不显著(P＞0.05)；两步法的鸡精液中精清和精子的甘油去除率极显著高其他两种方法(P＜0.01)；六步稀释法的精子回收率较高，但3种方法间无显著差异(P＞0.05)。在人工授精效果方面，不脱甘油组的受精率、活胚率和入孵蛋孵化率均为2.25%，极显著低于新鲜精液组和3种方法脱甘油组(P＜0.01)；3种脱甘油方法中六步稀释法的受精率、活胚率和入孵蛋孵化率相对较高，但与两步稀释法间无显著差异(P＞0.05)；两步稀释法的受精蛋孵化率近似新鲜精液，且活胚出苗率最高，达到了95.71%，但整体差异并不显著(P＞0.05)。【结论】使用1∶4两步稀释法能够高效去除鸡冷冻精液中的甘油，并获得良好的精子回收率、精子活力和人工授精效果。同时，相较于传统的六步稀释法，1∶4两步稀释法的操作流程更为简便，更适合在种鸡生产中应用。

关键词: 鸡冷冻精液; 脱甘油; 精子活力; 精子回收率; 受精率; 孵化率

Abstract: 【Objective】 This study was aimed to explored the influence of different glycerol removal methods on the artificial insemination efficacy of thawed chicken semen,aiming to simplify the glycerol removal procedure for chicken semen. 【Method】 Semen was collected from Wenchang roosters using the abdominal-dorsal combined massage method,frozen,thawed,and then diluted with extender at ratios of 1∶2,1∶4,1∶6,and 1∶8 (V/V) to remove glycerol.The optimal dilution ratio was determined by assessing sperm motility after deglycerolization.The optimal proportions were used for one-step,two-step,and six-step dilution methods for deglycerolization.The effectiveness of each method was evaluated by assessing sperm motility,sperm recovery rate,and glycerol content in both seminal plasma and sperm.Subsequently,the deglycerolized semen from the three methods was used for artificial insemination trials to measure fertilization and hatching rates. 【Result】 Among various dilution ratios,the 1∶4 six-step dilution method for deglycerolization demonstrated superior sperm motility both post-deglycerolization and at 30 minutes post-deglycerolization compared with 1∶2,1∶6,and 1∶8 six-step dilution methods,although the differences were not statistically significant (P＞0.05).Among different deglycerolization methods with a total dilution ratio of 1∶4,the six-step dilution method achieved the highest post-deglycerolization sperm motility at 68.34%,but the differences among the three methods were not significant (P＞0.05).The glycerol removal rates of both seminal plasma and sperm in the two-step dilution method were extremely significantly higher than those of the other two methods (P＜0.01).The sperm recovery rate was higher with the six-step dilution method,but no significant differences were found among the three methods(P＞0.05).In terms of artificial insemination outcomes,the fertilization rate,live embryo rate,and hatching rate of hatching eggs in non-glycerol-removed group were all 2.25%,which were extremely significantly lower than those of the fresh semen group and the three glycerol-removed method groups (P＜0.01).Among the three glycerol removal methods,the six-step dilution method showed relatively higher fertilization rate,live embryo rate,and hatching rate of hatching eggs,but no significant differences were observed compared with the two-step dilution method (P＞0.05).The hatching rate of fertilized eggs in the two-step dilution method was similar to that of fresh semen,and the live embryo emergence rate was the highest,reaching 95.71%,but the overall differences were not significant (P＞0.05). 【Conclusion】 The 1∶4 two-step dilution method efficiently removed glycerol from frozen chicken semen and achieved favorable sperm recovery rate,sperm motility,and artificial insemination outcomes.Meanwhile,compared with the traditional six-step dilution method,the 1∶4 two-step dilution method had a simpler operational process,making it more suitable for application in chicken breeding production.

Key words: chicken frozen semen; deglycerin removal; sperm vitality; sperm recovery rate; fertilization rate; hatching rate

中图分类号:

S814.3

何强, 蒋小香, 方婷婷, 李莉, 张守全, 卫恒习. 不同脱甘油方法对鸡冷冻精液人工授精效果的影响[J]. 中国畜牧兽医, 2025, 52(8): 3707-3714.

HE Qiang, JIANG Xiaoxiang, FANG Tingting, LI Li, ZHANG Shouquan, WEI Hengxi. Effect of Different Glycerol Removal Methods on Artificial Insemination of Cryopreserved Chicken Semen[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3707-3714.

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