甘肃某羊场绵羊流产衣原体的分离鉴定  

Isolation and identification of Chlamydia abortus in a sheep farm in Gansu

作　　者：李赟辉 李韦 张淮瑜 金有顺 赵鹏善 王彦俨 王一晗 肖乾 李兆才[2] 许信刚[1] 周继章[2] LI Yunhui;LI Wei;ZHANG Huaiyu;JIN Youshun;ZHAO Pengshan;WANG Yanyan;WANG Yihan;XIAO Qian;LI Zhaocai;XU Xingang;ZHOU Jizhang(College of Veterinary Medicine,Northwest A&F University,Yangling 712100,China;State Key Laboratory for Animal Disease Control and Prevention,College of Veterinary Medicine,Lanzhou University,Lanzhou Veterinary Research Institute,Chinese Academy of A gricultural Sciences,Lanzhou 730000,China;Gansu Yuansheng Farm Co.,Ltd.,Jingchang 737000,China)

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]中国农业科学院兰州兽医研究所动物疫病防控全国重点实验室,兰州大学动物医学与生物安全学院,甘肃兰州730000 [3]甘肃元生农牧科技有限公司,甘肃金昌737000

出　　处：《中国兽医科学》2024年第12期1601-1608,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2022YFC2304001);中国农业科学院科技创新工程项目(CAAS-ASTIP-2021-LVRI)。

摘　　要：为确定导致甘肃省某绵羊养殖场妊娠母羊流产的病原,笔者采集流产母羊的血清和胎水样本,采用ELISA、荧光PCR以及16S rRNA和ompA基因扩增测序,进行流产相关病原检测,并对阳性样本进行病原分离鉴定。结果显示,20份血清流产衣原体抗体均为阳性,而布鲁氏菌抗体和贝氏柯克斯体抗体均为阴性。5份胎水中仅检出流产衣原体特异性核酸。流产衣原体阳性胎水接种SPF鸡胚,成功分离到1株衣原体,经16S rRNA和ompA基因测序、包涵体观察和免疫荧光染色,鉴定分离株为流产衣原体。鸡胚半数致死量试验显示,0.4 mL流产衣原体分离菌株的ELD_(50)为10^(-7.55),表明分离株对鸡胚的毒力较强。结果表明,引起该绵羊养殖场妊娠绵羊流产的病原为流产衣原体,并成功分离到流产衣原体流行菌株,为绵羊流产衣原体病的流行病学和防制提供了参考。In order to identify the pathogen responsible for abortions in sheep in Gansu,serum and amniotic fluid samples were collected from aborted ewes and investigated.The abortion-related pathogens were detected using ELISA,fluorescent PCR,16S rRNA and omp A gene amplification followed by sequencing.The pathogen was further isolated and identified from positive samples.The results showed that all the 20 sera were positive for Chlamydia abortus antibody,while negative for Brucella and Coxiella burneti antibodies.In addition,Chlamydia abortus other than Brucella or Coxiella burneti specific nucleic acid was detected from the 5 amniotic fluid samples.A chlamydial strain was successfully isolated when the positive amniotic fluids were inoculated in SPF embryotic chicken eggs.The isolated strain was identified as Chlamydia abortus using 16S r RNA and omp A genes amplification and sequencing,inclusion body observation and immunofluorescence staining.Furthermore,the Chlamydia abortus isolate showed lethality to embryotic chicken egg with a ELD_(50)of 10^(-7.55)per 0.4 m L.Taken together,our results suggested that Chlamydia abortus may be the major pathogen responsible for the abortion storm in the sheep farm.The isolation and identification of the Chlamydia abortus endemic strain provided useful data and material for further study in epidemiology,prevention and controlling of abortions in sheep.

关 键 词：流产衣原体 分离鉴定 PCR 免疫荧光染色 绵羊 

分 类 号：S852.67[农业科学—基础兽医学]