摘要
旨在从广东某规模化鸡场死鸡胚进行鸡毒支原体(Mycoplasma gallisepticum,MG)的分离,并进行遗传进化分析、致病性及药物敏感性研究。本研究从广东某规模化鸡场的带菌死胚卵黄组织中分离禽支原体,通过菌落观察、血清学试验、16S rRNA支原体通用引物序列鉴定等方法,对分离的菌株进行种属鉴定,同时将分离的毒株对鸡胚和SPF鸡进行攻毒试验及抗菌药物敏感性试验。结果显示:显微镜下菌落呈典型的“荷包蛋”状。平板凝集试验显示其与MG阳性血清有凝集反应,与MS阳性血清不反应;16S rRNA序列测序发现各分离株与MG相似性达99.9%,因此确定分离株为MG。将各分离株感染7日龄SPF鸡胚,结果显示感染鸡胚大多于临近出壳时死亡;感染3周龄的SPF鸡,3周后解剖发现鸡气囊发生显著病理变化,说明其具有较强致病力;药物敏感性试验结果显示,5株分离株对盐酸沃尼妙林、多西环素、泰万菌素及泰妙菌素有较高敏感性,对替米考星、泰乐菌素、恩诺沙星、红霉素、吉他霉素、林可霉素呈现出不同程度耐药性。综上,从广东某规模化鸡场死鸡胚中成功分离到5株MG分离株,各分离株均可引起红细胞凝集,导致鸡胚死亡,对SPF鸡可引起典型的气囊炎症状,且对多种药物存在不同程度耐药性。本研究为临床MG的分离鉴定及用药选择提供了可参考的技术方法及指导,为MG攻毒模型建立提供了研究基础。
The aim of this study was to isolate Mycoplasma gallisepticum(MG)from dead chicken embryos in a large-scale chicken farm in Guangdong,and genetic evolution analysis,pathogenicity and drug susceptibility tests were carried out.Isolation of MG were conducted,then the isolated strains were identified by colony observation,serological test and 16S rRNA sequencing.At the same time,the isolated strains were used to infect chicken embryos and SPF chickens,and were subjected to antimicrobial susceptibility tests.Under the microscope,the colonies showed typical'poached egg'.Plate agglutination test showed that it had agglutination reaction with MG positive serum and did not react with MS positive serum.The 16S rRNA sequencing showed that the homology of each isolate with MG was 99.9%,so the isolates were identified as MG strains.Each isolate was used to infect 7-day-old SPF chicken embryos,and the results showed that most of the infected chicken embryos died near hatching.After 3 weeks of infection in 3-week-old SPF chickens,significant pathological changes were found in the air sac of chickens,indicating that it had strong pathogenicity.The drug sensitivity test showed that the 5 isolates were highly sensitive to warnimulin hydrochloride,doxycycline,tywanmycin and tymectin,and were resistant to tilmicosin,tylosin,enrofloxacin,erythromycin,kitasamycin and lincomycin to varying degrees.Five MG isolates were successfully isolated from dead chicken embryos in a large-scale chicken farm in Guangdong province.All of the isolates could cause chicken embryo death and cause typical air sac inflammatory symptoms to SPF chickens,and they had varying degrees of resistance to multiple drugs.This study provides reference technical methods and guidance for the isolation,identification and drug selection for clinical MG,and also provides a research basis for the establishment of MG challenge model.
作者
陈玥彤
刘晓涵
王芷洋
赵宇馨
周铁忠
胡增金
朱悦
王少辉
田明星
丁思羽
祁晶晶
于圣青
CHEN Yuetong;LIU Xiaohan;WANG Zhiyang;ZHAO Yuxin;ZHOU Tiezhong;HU Zengjin;ZHU Yue;WANG Shaohui;TIAN Mingxing;DING Siyu;QI Jingjing;YU Shengqing(Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200241,China;College of Animal Husbandry and Veterinary Medicine,Jinzhou Medicine University,Jinzhou 121000,China;Shandong Vocational Animal Science and Veterinary College,Weifang 261000,China;Yangzhou Youjiachuang Biotechnology Co.,Ltd,Yangzhou 225261,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2024年第1期290-299,共10页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(31902244)
中央级公益性科研院所基本科研业务费专项资金(2022JB05)
辽宁省教育厅基本科研项目(LJKMZ20221254)
扬州市市级计划-现代农业项目(YZ2022046)。
关键词
鸡毒支原体
死鸡胚
分离鉴定
致病性
药物敏感性
Mycoplasma galliscepticum
dead chicken embryos
isolation and identification
pathogenicity
drug susceptibility
