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雪貂(水貂)精液冷冻保存的比较以及经腹腔镜子宫内授精冻融精子后的妊娠情况

Comparative semen cryopreservation in ferrets (Mustela putorius furo) and pregnancies after laparoscopic intrauterine insemination with frozen-thawed spermatozoa.

作者信息

Howard J G, Bush M, Morton C, Morton F, Wentzel K, Wildt D E

机构信息

Department of Animal Health, National Zoological Park, Smithsonian Institution, Washington, DC 20008.

出版信息

J Reprod Fertil. 1991 May;92(1):109-18. doi: 10.1530/jrf.0.0920109.

DOI:10.1530/jrf.0.0920109
PMID:2056481
Abstract

A study was conducted to determine an optimum technique for semen cryopreservation and the biological competence of frozen-thawed ferret spermatozoa. Fifty-two fresh electroejaculates from 4 males were evaluated for sperm percentage motility, forward progressive motility, motility index (SMI) and acrosomal integrity. To determine the optimum temperature for maintaining sperm motility in vitro and the influence of glycerol on sperm motility, seminal aliquants were diluted in TEST diluent (containing either 0 or 4% glycerol) and maintained at 25 degrees or 37 degrees C. For cryopreservation, semen was diluted in each of 3 cryodiluents (TEST, PDV, BF5F), cooled for 30 min at 5 degrees C and pelleted on solid CO2 or frozen in 0.25 ml straws (20 degrees C/min to -100 degrees C). Following thawing, SMI and acrosomal integrity were determined. Ten females with maximum vulval swelling were given 90 i.u. human chorionic gonadotrophin and laparoscopically inseminated in utero with spermatozoa previously frozen using the optimum diluent and freeze-thaw method. The maintenance temperature of 25 degrees C was superior (P less than 0.05) to 37 degrees C for sustaining sperm motility, and glycerol did not influence (P greater than 0.05) motility for up to 11 h of culture. After thawing, motile spermatozoa were recovered in all treatment groups, but sperm motility and normal acrosomal ratings were highest using the PDV diluent, the pelleting method and thawing at 37 degrees C (P less than 0.05). Seven of the 10 ferrets (70%) inseminated with spermatozoa frozen by this approach became pregnant and produced 31 kits (mean litter size 4.4; range 1-9 kits).(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

开展了一项研究,以确定雪貂精液冷冻保存的最佳技术以及冻融后精子的生物学活性。对来自4只雄性雪貂的52份新鲜电刺激采得的精液进行了评估,测定精子的活率百分比、前向运动率、运动指数(SMI)和顶体完整性。为确定体外维持精子活力的最佳温度以及甘油对精子活力的影响,将精液样本用TEST稀释液(含0%或4%甘油)稀释,并分别置于25℃或37℃保存。进行冷冻保存时,精液分别用3种冷冻稀释液(TEST、PDV、BF5F)稀释,在5℃下冷却30分钟,然后置于固体二氧化碳上制成颗粒,或在0.25毫升细管中冷冻(以每分钟20℃的速度降至-100℃)。解冻后,测定SMI和顶体完整性。对10只外阴肿胀最大的雌性雪貂注射90国际单位人绒毛膜促性腺激素,然后通过腹腔镜将先前使用最佳稀释液和冻融方法冷冻的精子输入子宫。在维持精子活力方面,25℃优于37℃(P<0.05),甘油在长达11小时的培养过程中对精子活力无影响(P>0.05)。解冻后,所有处理组均能回收活动精子,但使用PDV稀释液、颗粒法和37℃解冻时,精子活力和正常顶体评级最高(P<0.05)。用这种方法冷冻的精子对10只雪貂中的7只(70%)进行授精,其中7只怀孕并产下31只幼崽(平均窝产仔数4.4只;范围为1-9只)。(摘要截短于250字)

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