鸽源ST443肺炎克雷伯菌的分离鉴定和生物学特性分析  

Isolation,Identification and Biological Characteristics Analysis of Pigeon Derived Klebsiella pneumoniae Strain ST443

作　　者：陈瑞格 李赟辉 项维 汪厚强 樊大艳[3] 李华明 雷连成 张付贤 CHEN Ruige;LI Yunhui;XIANG Wei;WANG Houqiang;FAN Dayan;LI Huaming;LEI Liancheng;ZHANG Fuxian(College of Animal Science and Technology,Yangtze University,Jingzhou 434023,China;Qianjiang Animal Husbandry Technology Promotion Station,Qianjiang 433136,China;Qianjiang Center for Animal Diseases Prevention and Control,Qianjiang 433100,China;College of Veterinary Medicine,Jilin University,Changchun 130062,China)

机构地区：[1]长江大学动物科学技术学院,湖北荆州434023 [2]潜江市畜牧技术推广站,湖北潜江433136 [3]潜江市动物疫病预防控制中心,湖北潜江433100 [4]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医杂志》2024年第5期46-55,共10页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金项目(32273006)。

摘　　要：为查明引起江苏省某养殖场赛鸽肺炎的病原及其生物学特性,本试验从患有呼吸道疾病的赛鸽肺组织中分离到1株肺炎克雷伯菌,通过形态学、生化鉴定、PCR扩增和16S rRNA基因测序对分离菌株进行鉴定,并采用多位点序列分型(MLST)、毒力基因检测、荚膜血清型分型、wzi基因测序、致病性分析、生物被膜形成能力测定、耐药基因检测、药敏试验和中药体外抑菌试验对分离菌株的生物学特性进行分析。结果显示,分离菌株为革兰阴性杆菌,显微镜下可见两端钝圆、杆状菌体,生化鉴定、khe基因PCR扩增和16S rRNA基因进化发育树分析确定其为肺炎克雷伯菌;MLST分型为ST443型;该菌株携带fimH、uge和wabG三种毒力基因;wzi基因测序鉴定分离菌株荚膜血清型为K16型;该菌株人工感染健康雏鸡可导致雏鸡多个器官出现不同程度的病变,半数致死量(LD_(50))为4.68×10^(9) CFU/mL;分离菌株具有中等生物被膜形成能力,携带bla SHV、bla TEM、tetM和sul 1四种耐药基因,对头孢曲松、头孢美唑、丁胺卡那、复方新诺明和多黏菌素敏感;中药明雄黄在体外对分离菌株有明显抑制作用。本试验为鸽源肺炎克雷伯菌感染的防控提供了参考。In order to identify the pathogen causing pneumonic disease in racing pigeons at a breeding farm in Jiangsu Province and elucidate its biological characteristics,one strain of Klebsiella pneumoniae was isolated from the lung tissue of pigeons suffering from respiratory diseases.The isolated strain was identified through morphological examination,biochemical tests,PCR amplification,and 16S rRNA gene sequencing.Further analysis of the biological characteristics of the isolated strain was conducted using multilocus sequence typing(MLST),virulence gene detection,capsular serotyping,wzi gene sequencing,pathogenicity analysis,biofilm formation ability determination,antibiotic resistance gene detection,drug sensitivity tests,and traditional Chinese medicine in vitro inhibition assays.The results showed that the isolated strain was a Gram-negative bacillus with blunt-rounded ends under the microscope,identified as Klebsiella pneumoniae through biochemical tests,khe gene PCR amplification,and 16S rRNA gene phylogenetic analysis.MLST typing revealed it belonged to ST443.The strain carried three virulence genes:fimH,uge,and wabG.Capsular serotyping based on wzi gene sequencing identified the strain as serotype K16.Inoculation of healthy chicks with the strain caused varying degrees of lesions in multiple organs,with a median lethal dose(LD_(50))of 4.68×10^(9) CFU/mL.The strain exhibited moderate biofilm formation ability and carried four antibiotic resistance genes:bla SHV,bla TEM,tetM,and sul 1.It was sensitive to ceftriaxone,cefmetazole,amikacin,compound sulfamethoxazole and polymyxin.In vitro,the traditional Chinese medicine realgar demonstrated significant inhibitory effects on the isolated strain.This study provides insights into the prevention and control of pigeon derived Klebsiella pneumoniae infection.

关 键 词：赛鸽 肺炎克雷伯菌 分离鉴定 ST443型 耐药性 

分 类 号：S836[农业科学—畜牧学]