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中国中部河南省奶牛场自然感染犬新孢子虫的发生情况及首次多位点微卫星基因分型

Occurrence and first multilocus microsatellite genotyping of Neospora caninum from naturally infected dogs in dairy farms in Henan, Central China.

作者信息

Qian Weifeng, Wang Tianqi, Yan Wenchao, Han Lifang, Zhai Kai, Duan Baoqing, Lv Chaochao

机构信息

College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, 471003, China.

Pulike Biological Engineering Co., Ltd., Luoyang, 471003, China.

出版信息

Parasitol Res. 2016 Aug;115(8):3267-73. doi: 10.1007/s00436-016-5142-y. Epub 2016 May 26.

Abstract

Neospora caninum is one of the important causes of abortion in dairy cattle worldwide. The dog is known as a definitive host of N. caninum and can transmit the parasite to cattle by shedding oocysts. The aim of the present study is to detect the presence of N. caninum in feces of dairy farm dogs and determine the genetic characteristics of N. caninum in Central China. A total of 78 fecal samples were collected from dogs in dairy farms from May to November 2014 and examined by microscopy and nested PCR based on Nc5 gene. Neospora-like oocysts were microscopically detected in two fecal samples, of which only one (Nc-LY1) was confirmed to be N. caninum by nested PCR. Seven out of 78 fecal samples (9.0 %) were N. caninum DNA positive, of which Neospora-like oocysts were simultaneously microscopically detected only in one sample (Nc-LY1). No statistical associations were found between the positive rates and age or sex of dogs (P > 0.05). The N. caninum-positive DNA samples were further analyzed by multilocus microsatellite (MS) genotyping for MS4, MS5, MS6A, MS7, MS8, MS10, MS12, and Cont-14. Only the fecal sample in which oocysts were detected was successfully genotyped at all genetic loci, and a new genotype was identified. To our knowledge, this study is the first report of genetic characterization of N. caninum isolates from naturally infected dogs based on multilocus microsatellites in China.

摘要

犬新孢子虫是全球范围内奶牛流产的重要病因之一。犬被认为是犬新孢子虫的终末宿主,可通过排出卵囊将该寄生虫传播给牛。本研究的目的是检测奶牛场犬粪便中犬新孢子虫的存在情况,并确定中国中部地区犬新孢子虫的遗传特征。2014年5月至11月,从奶牛场的犬只中总共采集了78份粪便样本,通过显微镜检查和基于Nc5基因的巢式PCR进行检测。在两份粪便样本中通过显微镜检测到类新孢子虫卵囊,其中只有一份(Nc-LY1)通过巢式PCR被确认为犬新孢子虫。78份粪便样本中有7份(9.0%)犬新孢子虫DNA呈阳性,其中仅在一份样本(Nc-LY1)中同时通过显微镜检测到类新孢子虫卵囊。在阳性率与犬的年龄或性别之间未发现统计学关联(P>0.05)。对犬新孢子虫DNA阳性样本进一步通过多位点微卫星(MS)基因分型检测MS4、MS5、MS6A、MS7、MS8、MS10、MS12和Cont-14。仅在检测到卵囊的粪便样本中所有基因位点成功进行了基因分型,并鉴定出一种新的基因型。据我们所知,本研究是中国基于多位点微卫星对自然感染犬的犬新孢子虫分离株进行遗传特征分析的首次报道。

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