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张敬,周军英,赵锡森,刘玉平.2014.白鹤冷冻精液人工授精实验.动物学杂志,49(6):925-929.
白鹤冷冻精液人工授精实验
Artificial Insemination with Cryopreserved Semen in Siberian Crane (Grus leucogeranus)
投稿时间:2013-12-06  修订日期:2014-10-15
DOI:DOI: 10.13859/j.cjz.201406017
中文关键词:  冷冻精液  稀释液  冷冻液  精子活率  白鹤
英文关键词:Frozen semen  Extender  Freezing solution  Rate of living sperm  Siberian crane (Grus leucogeranus)
基金项目:
作者单位E-mail
张敬 北京动物园 zhangjing0469@126.com 
周军英 北京动物园
中国动物园协会 
 
赵锡森 北京动物园  
刘玉平 北京动物园  
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中文摘要:
      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,同时在产完1枚卵后4 h内完成一次输精,效果最佳。
英文摘要:
      Cryogenic preservation 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 semen cryopreservation and the way to use it for artificial insemination of rare crane in captivity, a semen cryopreservation research on Siberian Crane (Grus leucogeranus) was undertaken at Beijing Zoo from 2003 to 2005. The semen of Siberian Crane was collected then preserved by freezing, the frozen semen was used subsequently for insemination. From fifteen times of 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, adjust pH to 7.8, and the osmotic pressure to 300 mosm/L (773 kPa) with distilled water, with 12% dimethylsulfoxide (DMSO) as the freezing solution to protection the sperms during cryopreservation. Semen is diluted with (3.2±1.1) times the volume of crane extender when it was collected. DMSO was added in the lab, the quantity of 12%DMSO was half volume of the diluted ejaculate, DMSO and diluted ejaculate were be at the same temperature (0-4℃) when they 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 into 0-4℃ water for 3-5 min, and the semen was inseminated immediately after thawing. The rate of living sperms after thawing was (29.3±15.5)%(n=16). Insemination of the Siberian crane with frozen semen was undertaken each day during egg laying period, and insemination within four hours after each laying egg as soon as possible, using the equivalent of two to four ejaculates for each insemination to make up the loss of sperms during freezing. Experament of Artificial insemination by frozen semen was undertook for two eggs of each cluth in 2004 and 2005, One egg was fertile from five eggs laid in two years, with one chick was bred successfully from frozen semen.
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