一株猪源A型多杀性巴氏杆菌的分离鉴定  

Isolation and identification of a strain of Pasteurella multocida type A from swine

作　　者：胡璇 蔡平 李红婷 温贵兰[1,2] 刘蔓蔓 孙芸 张小波 HU Xuan;CAI Ping;LI Hong-ting;WEN Gui-lan;LIU Man-man;SUN Yun;ZHANG Xiao-bo(College of Animal Science,Guizhou University,Guiyang 550025,China;Guizhou Engineering Research Center of Animal Biological Products,Guiyang 550016,China;Agriculture and Rural Affairs Bureau of Qianxi County,Guiyang 551500,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物生物制品工程技术研究中心,贵州贵阳550016 [3]贵州省黔西县农业农村局,贵州贵阳551500

出　　处：《中国兽医科学》2021年第12期1568-1578,共11页Chinese Veterinary Science

基　　金：国家自然科学基金/地区科学基金项目(32060794);贵州大学2020年省级教学内容和课程体系改革项目(GZJG-20200035);贵州省科技平台及人才团队计划项目(黔科合平台人才[2018]5253)。

摘　　要：2020年12月贵州毕节某猪场大批育肥猪发病死亡,为了鉴定导致其发病死亡的致病菌,无菌采集病死育肥猪病变内脏组织,采用细菌分离培养、16S rRNA PCR测序、荚膜分型、毒力基因检测、耐药基因检测、药敏试验、动物毒力试验等方法对分离菌株进行了病原学和分子生物学系统鉴定。细菌分离培养结果表明,分离菌在血平板上形成圆形、微突起、表面光滑、奶油状的菌落;分离菌16S rRNA核苷酸序列与NCBI中公布的多杀性巴氏杆菌的同源性达到99.9%以上,将分离菌命名为GZBJ2101;荚膜分型结果显示,GZBJ-2101株为荚膜A型多杀性巴氏杆菌;毒力基因检测结果显示,GZBJ2101株携带有hsf-1、pfhA、sodC等15种毒力基因;耐药基因检测结果显示,GZBJ2101株携带有β-内酰胺类、喹诺酮类、氯霉素类耐药基因;药敏试验结果显示,GZBJ2101株对米诺环素等10种药物敏感,对万古霉素等4种药物中度敏感,对多西环素等5种药物耐药。小鼠毒力试验显示GZBJ2101株具有较强的毒力。本试验成功从病死保育猪病变肺中分离到1株A型多杀性巴氏杆菌,研究结果可为养殖场多杀性巴氏杆菌的防控提供参考。In December 2020,a large number of fattening swine from certain swine farm died of a disease in Bijie,Guizhou Province.In order to identify the pathogenic bacteria that caused the disease and death,pathological splanchnic tissues of finishing swine were aseptically taken.Bacterial isolation and culture,16S rRNA PCR sequencing,capsular typing,virulence gene detection,drug resistance gene detection,drug sensitivity test,animal virulence test and other methods were used to systematically identify the pathogen and molecular biology.Isolation and culture results showed that the isolated bacteria formed round,micro-protruding,smooth and creamy colonies on the blood plate.The 16S rRNA nucleotide sequence of the isolated strain was more than 99.9%homology with Pasteurella multocida published by NCBI,and the isolated strain was named GZBJ2101.The results of capsular typing showed that the GZBJ2101 strain was P.multocida capsule type A.The results of virulence gene test showed that GZBJ2101 strain carried 15 virulence genes,including hsf-1,pfhA and sodC.The results of drug resistance gene test showed that GZBJ2101 strain carriedβ-lactams,quinolones and chloramphenicols resistance genes.The results of drug sensitivity test showed that GZBJ2101 strain was sensitive to 10 kinds of antibiotics such as minocycline,was moderately sensitive to 4 kinds of antibiotics such as vancomycin,and was resistant to 5 kinds of antibiotics such as doxycycline.Virulence test in mice showed that GZBJ2101 strain had strong virulence.A strain of P.multocida type A was successfully isolated from the diseased lungs of neonatal swine.The results can provide reference for the prevention and control of P.multocida in breeding farms.

关 键 词：多杀性巴氏杆菌 分离鉴定 毒力基因 

分 类 号：S852.612[农业科学—基础兽医学]