白鹤冷冻精液人工授精实验  

Artificial Insemination with Cryopreserved Semen in Siberian Crane (Grus leucogeranus)

作　　者：张敬[1] 周军英[1,2] 赵锡森[1] 刘玉平[1] 

机构地区：[1]北京动物园,北京100044 [2]中国动物园协会,北京100037

出　　处：《动物学杂志》2014年第6期925-929,共5页Chinese Journal of Zoology

摘　　要：为了探索鹤类精液冷冻保存和使用技术,2003~2005年,进行了白鹤（Grus leucogeranus）的冷冻精液保存及人工授精实验。使用Beltsville家禽精液稀释液作为白鹤精液稀释液,12%的二甲基亚砜（DMSO）为冷冻液。精液样本冷冻经过三个阶段的降温,最后保存在液氮中。成功保存了编号93001雄性白鹤精液36支（0.2 ml/支）。冷冻精液在0~4℃冰水中解冻3~5 min,解冻后白鹤精液精子活率为29.3%±15.5%（n=16）,2004和2005年分别为92101号雌鹤产的两窝卵进行人工授精实验,2年共产卵5枚,其中1枚卵受精并成功孵化。实验发现在雌鹤产卵前一周和产卵期间每天输精,并增加每次输精量,同时在每产完1枚卵后4 h内完成一次输精,效果最佳。Cryopreservation of semen is a useful technology for maximizing fertility and genetic diversity when breeding endangered species in captivity. In order to explore the technology of artificial insemination of rare crane in captivity,a semen cryopreservation test in Siberian Crane（ Grus leucogeranus） was conducted at Beijing Zoo from 2003 to 2005. The semen of Siberian Crane was collected and preserved by freezing,and the frozen semen was used subsequently for insemination. From fifteen semen collections,thirty-six 0. 2 ml samples were frozen. Frozen semen was thawed for AI sixteen times. The Beltsville Poultry Semen Extender was used.We adjusted p H to 7. 8,and the osmotic pressure to 300 mosm / L（ 773 k Pa） with distilled water,and used12% dimethylsulfoxide（ DMSO） as the freezing solution to protect the spermatozoa from damage during cryopreservation. Semen was diluted 3. 2 ± 1. 1 times of the volume of crane extender when collected. 12%DMSO was added,in half volume of the diluted ejaculate,and DMSO and diluted ejaculate were at the same temperature（ 0- 4℃） when they were mixed. The ejaculate was equilibrated with DMSO for 15 min at 0- 4℃in the freezer. The samples were put into 0. 2 ml each in freezing tubes for cryopreservation. Frozen semen was kept in the liquid nitrogen following three temperature-fall steps： 4-- 20℃,- 20-- 80℃,- 80-- 196℃. Frozen semen was thawed in 0- 4℃ water for 3- 5 min,and the semen was used for insemination immediately after thawing. The rate of living spermatozoa after thawing was 29. 3% ± 15. 5%（ n = 16）.Insemination of the Siberian Crane with frozen semen was conducted each day during egg laying period,withinfour hours after egg laying,using the equivalent of two to four ejaculates to make up the loss of spermatozoa during freezing. Artificial insemination by frozen semen was conducted for two clutches of eggs in 2004 and2005. One egg was fertilized among five eggs laid in two years,and one chick was successfully bred from frozen semen.

关 键 词：冷冻精液 稀释液 冷冻液 精子活率 白鹤 

分 类 号：Q492.6[生物学—生理学]