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渔猫(Prionailurus viverrinus)基本精液特征、精子冷冻保存及异种体外受精的评估

Assessment of basic seminal characteristics, sperm cryopreservation and heterologous in vitro fertilisation in the fishing cat (Prionailurus viverrinus).

作者信息

Thiangtum Khongsak, Swanson William F, Howard JoGayle, Tunwattana Wanchai, Tongthainan Dakara, Wichasilpa Wisid, Patumrattanathan Pornchai, Pinyopoommintr Tanu

机构信息

Center of Agricultural Biotechnology, Kasetsart University, Kamphaengsaen Campus, Nakhon Pathom 73140, Thailand.

出版信息

Reprod Fertil Dev. 2006;18(3):373-82. doi: 10.1071/rd05098.

DOI:10.1071/rd05098
PMID:16554013
Abstract

Conservation of the fishing cat, a threatened south-east Asian felid, could benefit from effective ex situ genetic management and breeding programmes, including the use of assisted reproduction. The aims of the present study were to: (1) characterise basal seminal traits of fishing cats in Thailand zoos; and (2) investigate the effect of cryopreservation on sperm motility, acrosomal integrity and in vitro function. Seminal traits were evaluated in electroejaculates collected from eight males. Spermatozoa were diluted in n-tris(hydroxymethyl)-methyl-2-aminoethanesulfonic acid Tris (TEST)-yolk buffer (TYB) without glycerol, then diluted further with TYB with glycerol (4% final concentration) at either 25 degrees C or after slow cooling to 5 degrees C and frozen in straws over liquid nitrogen vapour. After thawing, sperm function was assessed by insemination of viable domestic cat oocytes. Fishing cat ejaculates averaged (+/- s.e.m.) 43.6 +/- 14.2 x 10(6) motile spermatozoa with 33.5 +/- 6.8% normal sperm morphology. Semen processing had a negligible effect (P > 0.05) on sperm motility and acrosomal integrity, but values were reduced (P < 0.05) after thawing. All thawed samples fertilised domestic cat oocytes, with 62.1% (36/58) of mature oocytes cleaving. Glycerol addition at 5 degrees C resulted in higher (P < 0.05) post-thaw motility and intact acrosomes than glycerol addition at 25 degrees C. In conclusion, good-quality ejaculates can be obtained from Thai fishing cats and their spermatozoa exhibit adequate function after cryopreservation for in vitro fertilisation procedures.

摘要

渔猫是一种濒危的东南亚猫科动物,对其进行保护可受益于有效的迁地遗传管理和繁殖计划,包括使用辅助生殖技术。本研究的目的是:(1)描述泰国动物园中渔猫的基础精液特征;(2)研究冷冻保存对精子活力、顶体完整性和体外功能的影响。对从8只雄性渔猫采集的电刺激射精精液的特征进行了评估。精子在不含甘油的n-三(羟甲基)甲基-2-氨基乙磺酸三羟甲基氨基甲烷(TEST)-卵黄缓冲液(TYB)中稀释,然后在25℃或缓慢冷却至5℃后,用含甘油(终浓度4%)的TYB进一步稀释,并在液氮蒸汽上冷冻于细管中。解冻后,通过对有活力的家猫卵母细胞进行授精来评估精子功能。渔猫射精平均(±标准误)有43.6±14.2×10⁶条活动精子,正常精子形态占33.5±6.8%。精液处理对精子活力和顶体完整性的影响可忽略不计(P>0.05),但解冻后这些值有所降低(P<0.05)。所有解冻样本均使家猫卵母细胞受精,62.1%(36/58)的成熟卵母细胞发生分裂。在5℃添加甘油比在25℃添加甘油解冻后的活力和完整顶体率更高(P<0.05)。总之,可从泰国渔猫获得高质量的射精精液,其精子在冷冻保存后表现出足够的功能用于体外受精程序。

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