河北省5株牛支原体的分离鉴定和药敏试验  

Isolation, Identification, and Drug Sensitivity Test of Five Mycoplasma bovis Strains in Hebei Province

作　　者：张锦程 白和平 赵桂新 王少华[3] 郭建军 吴同垒[2] 史秋梅[2] ZHANG Jincheng;BAI Heping;ZHAO Guixin;WANG Shaohua;GUO Jianjun;WU Tonglei;SHI Qiumei(Donggang Animal Disease Prevention and Supervision Station,Harbour District,Qinhuangdao City,Qinhuangdao 066000,China;Key Laboratory of Preventive Veterinary Medicine in Hebei Province,College of Animal Science and Technology,Hebei Normal University of Science and Technology,Qinhuangdao 066604,China;College of Animal Science and Technology,HeBei North University,Zhangjiakou 075131,China;Chengde Academy of Agriculture and Forestry,Chengde 067500,China)

机构地区：[1]秦皇岛市海港区东港动物防疫监督站,河北秦皇岛066000 [2]河北科技师范学院动物科技学院河北省预防兽医学重点实验室,河北秦皇岛066604 [3]河北北方学院动物科技学院,河北张家口075131 [4]承德市农林科学院,河北承德067500

出　　处：《中国兽医杂志》2024年第6期64-69,共6页Chinese Journal of Veterinary Medicine

基　　金：河北省农业厅项目(HBCT2024240201);中央引导地方科技发展资金资助项目(236Z6604G);河北省教育厅青年基金项目(QN2017305);围场御道口智慧、循环农牧业示范建设区项目(202202F016)。

摘　　要：为了确定肉牛场犊牛患呼吸道疾病死亡的致病原,本试验采集河北省秦皇岛地区5个肉牛养殖场病牛的肺脏组织进行病原的分离培养,采用染色镜检、培养特性观察、支原体特异性oppD/F基因鉴定、16S rRNA序列比对和同源性分析鉴定分离菌株,并通过药敏试验筛选敏感药物,检测中西药对分离菌株的抑菌效果。结果显示,分离菌株在PPLO肉汤固体基础培养基上为针尖大小白色菌落,瑞氏染色菌体呈球状;oppD/F基因和16S rRNA基因扩增均可得预期目的条带,16S rRNA基因序列与NCBI中牛支原体参考序列同源性达99%,在遗传进化树中处于同一分支,共从病牛肺脏分离鉴定出5株牛支原体,分别命名为ZYT-1~ZYT-5;药敏试验结果显示,5株牛支原体分离菌株对恩诺沙星、环丙沙星、土霉素和氟苯尼考极度敏感,对中药苏木、五味子、白头翁和乌梅极度敏感。结果表明,肉牛场患病犊牛的致病原为牛支原体,本试验结果可为当地牛支原体感染临床病例的治疗用药提供参考依据。To identify the pathogen responsible for respiratory disease deaths in calves at beef cattle farms,lung tissue samples were collected from five diseased cattle at beef cattle farms in the Qinhuangdao of Hebei Province.Pathogens were isolated and cultured,and identified through staining microscopy,observation of cultural characteristics,Mycoplasma specific oppD/F gene identification,16S rRNA sequence alignment,and homology analysis.Sensitivity tests were conducted to screen for effective antibiotics,assessing the antibacterial effects of both Western and traditional Chinese medicines on the isolated strains.The results showed that the isolated strains formed white pinpoint-sized colonies on PPLO solid medium and appeared spherical under Wright's staining.Both oppD/F gene and 16S rRNA gene amplifications yielded expected bands,and the 16S rRNA gene sequence showed 99%homology with the Mycoplasma bovis reference sequences in NCBI.Five Mycoplasma bovis strains were identified and named ZYT-1 to ZYT-5.Antibiotic sensitivity tests indicated that the five Mycoplasma bovis isolates were highly sensitive to enrofloxacin,ciprofloxacin,oxytetracycline,and florfenicol,as well as to traditional Chinese medicines such as Sappanwood,Schisandra,Pulsatilla,and Mume.These findings suggest that the pathogen causing the disease in calves at beef cattle farms is Mycoplasma bovis.The results of this study can provide a reference for the treatment of clinical cases of Mycoplasma bovis infection in the region.

关 键 词：牛支原体 瑞氏染色 16S rRNA序列比对 特异基因检测 药敏试验 

分 类 号：S858.23[农业科学—临床兽医学]