1株山鸡源致病性大肠杆菌的分离鉴定  

Isolation and identification of pathogenic Escherichia coli from pheasant

作　　者：张春晓 赵玉林 杨富琳 王利丽 赵奇 侯冠欣 孙欣艺 张志强[1] Zhang Chunxiao;Zhao Yulin;Yang Fulin;Wang Lili;Zhao Qi;Hou Guanxin;Sun Xinyi;Zhang Zhiqiang(Animal Science and Technology College,Hebei Normal University of Science&Technology,Hebei Key Laboratory of Preventive Veterinary Medicine,Hebei Qinhuangdao 066000;Innovation Technology Center of HAID Research Institute,Guangdong HAID Group Co.,Ltd.,Guangdong Guangzhou 510660)

机构地区：[1]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066000 [2]海大研究院创新技术中心,广东海大集团股份有限公司,广东广州510660

出　　处：《现代畜牧兽医》2024年第1期1-7,共7页Modern Journal of Animal Husbandry and Veterinary Medicine

基　　金：国家自然科学基金资助项目(31802186);河北省高等学校科学技术研究项目(BJ2021055);河北省重点研发计划项目(19226629D)。

摘　　要：为查明秦皇岛市某山鸡养殖场山鸡大批量死亡病因,试验对分离到的致病优势菌株进行生化鉴定和16S rRNA基因序列分析,并测试分离菌对21种抗生素的敏感性,进一步采用PCR方法检测分离菌的耐药基因。利用清洁级昆明小鼠和SPF鸡胚分别对分离菌进行致病性试验,并采用PCR方法检测分离菌的毒力基因。结果显示:分离菌为革兰氏阴性杆菌,在伊红美蓝培养基上生长出有金属光泽的菌落,生化特性与大肠杆菌生化特性符合率高达99%。BLAST分析发现,分离菌16S rRNA序列与大肠杆菌的基因相似性高达99.8%。分离菌对恩诺沙星和阿米卡星高度敏感,对青霉素、头孢曲松等15种药物耐药。分离菌的磺胺类、β-内酰胺类、四环素类的耐药基因与耐药表型基本相符,其余耐药基因与耐药表型不符,提示该分离菌可能存在其他的耐药机制。致病性试验结果显示,分离菌具有较强致病性,共检测到12种大肠杆菌主要毒力基因。研究表明,分离菌为山鸡源致病性大肠杆菌,且耐药情况复杂,具有较强的毒力和致病性。In order to investigate the cause of mass deaths in a pheasant farm in Qinhuangdao City,the isolated dominant pathogenic strains were subjected to biochemical identification and 16S rRNA gene sequence analysis.Additionally,the sensitivity of the isolated bacteria to 21 antibiotics was tested,and PCR was employed to detect antibiotic resistance genes.Pathogenicity tests were conducted using clean Kunming mice and SPF chicken embryos,and PCR was used to detect virulence genes of the isolated bacteria.The results revealed that the isolated bacteria were Gram-negative rods,forming colonies with a metallic sheen on eosin methylene blue agar,and their biochemical characteristics matched those of Escherichia coli with a high agreement rate of 99%.BLAST analysis indicated a 99.8%similarity between the 16S rRNA sequence of the isolated bacteria and that of Escherichia coli.The isolated bacteria exhibited high sensitivity to enrofloxacin and amikacin but resistance to 15 other drugs,including penicillin and ceftriaxone.The resistance genes of the isolated bacteria related to sulfonamides,β-lactams,and tetracyclines were consistent with the resistance phenotype,while other resistance genes did not align with the resistance phenotype,suggesting the possible existence of other resistance mechanisms.Pathogenicity tests showed that the isolated bacteria had strong pathogenicity,and 12 major virulence genes of Escherichia coli were detected.The study indicates that the isolated bacteria are pathogenic Escherichia coli from pheasant,exhibiting complex antibiotic resistance and strong virulence.

关 键 词：山鸡源致病性大肠杆菌 药敏试验 毒力基因 耐药基因 致病性试验 

分 类 号：S852.61[农业科学—基础兽医学]