鸭源多杀性巴氏杆菌的分离鉴定及耐药性分析  

Isolation,Identification and Drug Resistance Analysis of Pasteurella multocida from Ducks

作　　者：陈国权 张旭[1,2] 阎朝华 周碧君 王开功[1,2,3] 程振涛 文明[1,2,3] CHEN Guoquan;ZHANG Xu;YAN Chaohua;ZHOU Bijun;WANG Kaigong;CHENG Zhentao;WEN Ming(College of Animal Science,Guizhou University,Guiyang 550025,China;Institute of Animal Disease,Guizhou University,Guiyang 550025,China;Key Laboratoryof Animal Diseases and Veterinary Public Health of Guizhou Province,Guiyang 550025,China)

机构地区：[1]贵州大学动物科学学院,贵阳550025 [2]贵州大学动物疫病研究所,贵阳550025 [3]贵州省动物疫病与兽医公共卫生重点实验室,贵阳550025

出　　处：《中国畜牧兽医》2020年第2期620-628,共9页China Animal Husbandry & Veterinary Medicine

基　　金：贵州省百层次创新型人才项目(黔科合人才[2016]4009号);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号);三穗鸭工程技术研究中心建设项目(黔科合平台人才[2019]5203号)

摘　　要：为鉴定临床疑似鸭多杀性巴氏杆菌感染肉鸭的病原菌,本试验通过细菌分离培养、菌体形态观察、细菌生化鉴定、16S rRNA基因测序分析、细菌种特异性鉴定、荚膜分型鉴定和动物回归试验进行鉴定,并通过药敏试验和耐药基因检测进行耐药性分析。结果显示,从患病鸭肝脏组织分离到的细菌在鲜血琼脂培养基中呈现表面光滑凸起、灰白色菌落,为革兰氏阴性短小杆菌,瑞氏染色呈两极浓染;生化鉴定结果显示,分离菌能发酵葡萄糖、蔗糖和甘露醇,硫化氢、氧化酶和吲哚等试验阳性;16S rRNA基因序列系统进化树分析显示,该分离菌与多杀性巴氏杆菌聚为一支,同源性>99%;细菌种特异性鉴定结果与多杀性巴氏杆菌相符;荚膜分型鉴定结果仅扩增到约为1050 bp的目的基因片段,与荚膜血清A型相符;动物回归试验显示,该分离菌有较强的致病性;药敏试验结果显示,该分离菌对羧苄西林、氨苄西林、复方新诺明和四环素等12种药物耐药;经耐药基因PCR检测显示,该分离菌携带Sul1、Sul3、tet(X)和Intl 14种耐药基因,与药敏表型相符。本试验成功分离到1株鸭源荚膜血清A型多杀性巴氏杆菌,为鸭多杀性巴氏杆菌病的防治提供参考依据。In order to identify the pathogens of clinically suspected duck Pasteurella multocida infection,in this experiment,bacteria isolated and cultured,morphological observation,biochemical identification,16S rRNA gene sequencing analysis,bacterial species-specific identification,capsule type identification and animal regression test were carried out,drug resistance was analyzed by drug sensitivity test and drug resistance gene detection.The results showed that the bacteria isolated from the liver tissue of the diseased duck showed smooth bulging and gray-white colonies on the blood agar medium,which was Gram-negative bacillus,and the Wright’s staining showed two-level thick staining;Biochemical identification showed that the isolated strain could ferment glucose,sucrose and mannitol,and the test results of hydrogen sulfide,oxidase and hydrazine were positive.The phylogenetic tree analysis of the 16S rRNA gene sequence showed that the isolated strain was clustered with the genus Pasteurella,homology>99%.The results of bacterial species specific identification were consistent with Pasteurella multocida.The results of capsule typing only amplified the target gene fragment of about 1050 bp,which was consistent with capsule serotype A.Animal regression test showed that the isolated strain had strong pathogenicity.The drug sensitivity test showed that the isolated strain was resistant to 12 drugs such as carbenicillin,ampicillin,cotrimoxazole and tetracycline.PCR detection of drug resistance gene showed that the isolate carried Sul1,Sul3,tet(X)and Intl 1 four drug resistance genes,consistent with the drug sensitivity phenotype.In this experiment,a duck capsular serotype A Pasteurella multocida was successfully isolated,which provided a reference for the prevention and treatment of duck Pasteurella multocida infection.

关 键 词：鸭 多杀性巴氏杆菌 分离鉴定 耐药性 

分 类 号：S852.61[农业科学—基础兽医学]