秦岭大熊猫精液的细管冷冻保存

Straw Cryopreservation of Semen of Giant Panda from Qinling Mountains

2008年3月1日至4月27日和2009年3月3日至5月1日,在陕西省珍稀野生动物抢救饲养研究中心对处于繁殖期内的4只雄性秦岭大熊猫(Ailuropoda melanoleuca qinlingensis)精液进行了细管冻精实验。比较组成不同的4种稀释液:葡萄糖-果糖-柠檬酸三钠-卵黄-甘油-双抗(稀释液1)、葡萄糖-蔗糖-柠檬酸三钠-卵黄-甘油-双抗(稀释液2)、葡萄糖-柠檬酸三钠-卵黄-甘油-双抗(稀释液3)和美国进口的TEST(加入3.5%甘油),以及直接降温平衡法(方法 1)与逐级降温平衡法(方法 2)2种冷冻保存操作方法,对秦岭大熊猫精液进行细管冷冻保存后精子活力和顶体完整率的影响。结果表明:稀释液1的精子活力为46.25%±11.67%,顶体完整率为80.75%±7.89%,TEST的精子活力为48.75%±8.54%,顶体完整率为84.50%±7.59%,两者的精子活力和顶体完整率均无明显差异(P>0.05),但是都明显高于稀释液2(P﹤0.01)和稀释液3(P﹤0.01);采用方法 1冷冻保存秦岭大熊猫精液,解冻后精子的活力和顶体完整率分别为45.67%±10.54%和81.37%±8.42%,都显著高于方法 2(P﹤0.01);方法 1解冻后畸形率为23.50%±3.51%,明显低于方法 2(P﹤0.01)。经比较确定,方法 1(用稀释液1)是一种较好的细管冷冻保存秦岭大熊猫精液的方法。

From March 1 st to April 27th, 2008 and from March 3rd to May 1 st, 2009, the semen samples from four breeding Giant pandas which distributed in Qinling Mountains （Ailuropoda melanoleuca qinlingensis） were used for semen cryopreservation experiments in Shaanxi Rare Wildlife Reseue and Breeding Research Center. The sperm motility and acrosome integrity were tested after straw cryopreservation of semen with 4 diluents, glucose-fructose-trisodium citrate-yolk-glycerin-antibiotics, glucose-sucrose-trisodium citrate-yolk-glycerin- antibiotics, glucose-trisodium citrate-yolk-glycerin-antibiotics, and TEST imported form the USA （added with 3.5% glycerin）, and with 2 methods, direct cooling balance method, and gradual cooling balance method. The results showed that the percentages of sperm motility and intact aerosome were 46.25% ± 11.67% and 80.75% ±7.89% respectively in diluent 1 ; 48.75% ±8.54% and 84.50% ±7.59% in diluent TEST. There was no significant difference between these two diluents （ P 〉 0.05 ）, but both were better than diluent 2 and diluent 3 （P 〈0.01 ）. After freezing and thawing, the percentages of sperm motility and intact acrosome were 45.67% ± 10.54% and 81.37% ± 8.42% respectively in cryopreservation method 1, significantly higher than those of method 2 （ P 〈 0.01 ）. The percentage of sperm abnormality was 23.50% ± 3.51% , in method 1, significantly lower than that of method 2 （P 〈 0.01 ）. Thus, we conclude that method 1 （ with diluent 1 ） is good for semen straw cryopreservation in Giant Panda.