山羊肺源肠外致病型大肠杆菌的分离鉴定、耐药性测定和毒力基因检测  

Isolation,Identification,Drug Resistance and Virulence Gene Detection of Extraintestinal Pathogenic Escherichia coli from Goat Lung

作　　者：徐海军[1] 李小红 爨淑楠 张娅菲 贾哲 王书明 张圣尧 XU Haijun;LI Xiaohong;CUAN Shunan;ZHANG Yafei;JIA Zhe;WANG Shuming;ZHANG Shengyao(College of Biological and Pharmaceutical Engineering,West Anhui University,Lu’an Anhuui 237012,China;Anhui Jinfengyuan Animal Husbandry Technology Co.,Ltd,Lu’an Anhui 237000,China)

机构地区：[1]皖西学院生物与制药工程学院,安徽六安237012 [2]安徽金丰源畜牧科技有限公司,安徽六安237000

出　　处：《西北农业学报》2023年第2期187-197,共11页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：安徽省高校优秀拔尖人才培育项目(gxgnfx2019026);皖西学院高层次人才科研启动资助项目(WGKQ201802001);安徽省本科质量工程项目(2020szsfkc0908);皖西学院校级本科质量工程项目(wxxy2020056)。

摘　　要：为鉴定一例山羊肺炎的细菌性病原,对病羊进行病理剖检观察,并从肺脏中分离鉴定细菌,对分离株进行溶血试验、系统进化群分析、致病性测定、毒力基因检测、耐药性检测,根据药敏试验结果选择敏感药物对患病山羊进行治疗,对全群山羊给药预防。结果发现,病羊肺上半部分散在不规则红色肝变病灶、心肌柔软;肺病变组织接种后分离到40个灰白色菌落,结合革兰氏染色镜检形态特点、选择性培养基上菌落形态特征及16S rRNA基因序列分析,鉴定为大肠杆菌;系统进化群分析结果表明,该菌属于B1群;致病性测定结果表明,该菌对小鼠的致死率为80%;毒力基因检测结果表明,该菌携带8种毒力基因(ituA、afa、tsh、ompA、fimH、gafD、iucD、traT);药敏试验结果显示,该菌对头孢曲松、氟苯尼考、恩诺沙星和阿米卡星中度敏感,对卡那霉素等9种抗菌药耐药。经用头孢噻呋钠和麻杏石甘散治疗后,羊群病情得到控制。To find out a bacterial pathogen of goat pneumonia,a pathological observationon the diseased goats was carried out and the bacteria were isolated from the lungs and identified by 16S rRNA sequencing.The isolated strains were characterized by assays related to hemolysis,phylogenetic tree,pathogenicity,virulence gene and drug resistance.Based on the antibiotic sensitive result,the effective antibiotic was selected to treat the diseased goats,and the whole group of goats was administrated to prevent.Pathological examination showed that irregular red liver-like lesions scattered in the upper part of the lung and myocardium was soft.Forty gray white colonies were isolated from the diseased lung tissue after inoculation.Combined with Gram staining characterization,colonial morphologyidentified by selection medium and 16S rRNA sequencing,the isolates were identified as Escherichia coli.Phylogenetic analysis showed that the strain belonged to group B1.Pathogenicity test showed that the lethal rate of the strain to mice was 80%.Virulence gene test showed that the strain carried eight virulence genes including ituA,afa,tsh,ompA,fimH,gafD,iucD,traT.Drug sensitivity test showed that the strain was moderately sensitive to ceftriaxone,florfenicol,enrofloxacin and amikacin,and resistant to other 9 kinds of antibiotics such as kanamycin.After treatment with ceftiofur sodium and Maxing Shigan powder,the goat’s epidemic situation had been controlled.These results can provide a reference for the clinical diagnosis and treatment of goat pneumonia caused by extraintestinal pathogenic E.coli.

关 键 词：山羊 肺炎 大肠杆菌 药敏试验 致病力试验 毒力基因 

分 类 号：S852.61[农业科学—基础兽医学]