河南郑州地区奶牛源安氏隐孢子虫虫种鉴定及多位点序列分型研究  

Identification and multilocus sequence typing of Cryptosporidium andersoni from dairy cows in Zhengzhou,Henan Province

作　　者：郑双健 李东方 陈远才 曹乐天 王璐阳 黄建营 刘东海 李豪 张龙现 ZHENG Shuang-jian;LI Dong-fang;CHEN Yuan-cai;CAO Le-tian;WANG Lu-yang;HUANG Jian-ying;LIU Dong-hai;LI Hao;ZHANG Long-xian(International Joint Research Laboratory for Zoonotic Diseases of Henan Province,College of Animal Science and Veterinary Medicine,Henan Agricultural University,Zhengzhou 450002,China;Henan Dairy Production Performance Testing Co.,Ltd.,Zhengzhou 450046,China)

机构地区：[1]河南农业大学牧医工程学院,河南省人兽共患病国际联合实验室,河南郑州450046 [2]河南省奶牛生产性能测定有限公司,河南郑州450046

出　　处：《中国兽医学报》2020年第9期1783-1790,共8页Chinese Journal of Veterinary Science

基　　金：河南省自然科学基金重点资助项目(162300410129)。

摘　　要：为了解河南省郑州地区奶牛隐孢子虫及安氏隐孢子虫(Cryptosporidium andersoni,C.andersoni)的亚型分布情况,本研究共采集3个奶牛场共973份奶牛新鲜粪便样品,采用显微镜检测方法进行隐孢子虫检测,对形态学鉴定的8份C.andersoni阳性样品分别基于SSU rRNA、Actin、HSP70和COWP基因位点进行虫种鉴定,基于CM-MS1、CM-MS2、CM-MS3和CM-MS16基因位点进行多位点序列分型(multilocus sequence typing,MLST)分析。结果显示,隐孢子虫总感染率为2.77%(27/973),而且以断奶前犊牛感染率最高7.10%(12/169)。8份样品在不同基因位点均成功鉴定为C.andersoni分离株,并形成4个MLST亚型,其中A4、A4、A4、A1为优势亚型,A6、A4、A2、A1为新亚型,且不同亚型的卵囊大小具有显著差异。研究表明,河南省郑州地区奶牛隐孢子虫总体感染水平较低,但是对奶牛养殖业和牛奶产量仍然有较大影响,此外,C.andersoni存在多个MLST亚型,表明具有遗传多样性。To understand the distribution of Cryptosporidium and Cryptosporidium andersoni subtypes in dairy cattles in Zhengzhou,Henan Province,a total of 973 fresh dairy cattle feces were collected from three dairy farms in this study.All samples were examined by microscopy to detect Cryptosporidium oocysts,and 8 C.andersoni positive samples identified by morphology were identified based on SSU rRNA,Actin,HSP70 and COWP loci,and then genotyped by multilocus sequence typing based on CM-MS1,CM-MS2,CM-MS3,and CM-MS16 loci.The results showed that the total infection rate of Cryptosporidium was 2.77%(27/973),and preweaned calves infection was most serious at 7.10%(12/169).Eight samples were all successfully identified as C.andersoni positive and genotyped at all loci,and classified into 4 MLST subtypes.Among them,A4,A4,A4,A1 were dominant MLST subtype,and A6,A4,A2,A1 were new MLST subtype,and there were significant differences in oocyst size between different subtypes.This study showed that the overall infection level of Cryptosporidium in dairy cattles in Zhengzhou,Henan Province was low,but it still had a great impact on dairy farming and milk yield.In addition,C.andersoni had several different MLST subtypes,indicating genetic diversity.

关 键 词：河南 奶牛 安氏隐孢子虫 多位点序列分型 

分 类 号：S852.7[农业科学—基础兽医学] R535[农业科学—兽医学]