甘肃某羊场绵羊流产衣原体的分离鉴定OA北大核心CSTPCD
Isolation and identification of Chlamydia abortus in a sheep farm in Gansu
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16SrRNA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16SrRNA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ELD50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 ompA 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 Cox-iella burneti antibodies.In addition,Chlamydia abortus other than Brucella or Coxiella burneti spe-cific 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 rRNA and ompA genes amplification and sequencing,inclusion body observation and immunofluorescence staining.Furthermore,the Chlamydia abortus iso-late showed lethality to embryotic chicken egg with a ELD50 of 10-7 55 per 0.4 mL.Taken together,our re-sults 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 use-ful data and material for further study in epidemiology,prevention and controlling of abortions in sheep.
李赟辉;许信刚;周继章;李韦;张淮瑜;金有顺;赵鹏善;王彦俨;王一晗;肖乾;李兆才
西北农林科技大学动物医学院,陕西杨凌 712100||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西北农林科技大学动物医学院,陕西杨凌 712100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甘肃元生农牧科技有限公司,甘肃金昌 737000
畜牧业
流产衣原体分离鉴定PCR免疫荧光染色绵羊
Chlamydia abortusisolation and identificationPCRimmunofluorescence stainingsheep
《中国兽医科学》 2024 (012)
1601-1608 / 8
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)
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