盘羊源多杀性巴氏杆菌与溶血性曼氏杆菌的分离与鉴定

Isolation and identification of Pasteurella multocida and Mannerella hemolytica from argali

为探究导致甘肃省某动物园盘羊死亡的病原菌及其生物学特性,笔者采集1只出现剧烈咳嗽、呼吸困难,经救治无效死亡的盘羊的肺组织进行细菌分离培养,通过细菌形态学观察和16S rRNA基因测序鉴定细菌种类,并对分离菌株进行荚膜血清型鉴定、毒力基因检测、药敏试验以及对小鼠的致病性试验。结果显示,从盘羊肺中分离鉴定出1株强致病性的D型多杀性巴氏杆菌和1株强致病性的A2型溶血性曼氏杆菌。D型多杀性巴氏杆菌分离株携带exbB、exbD、fur、tonB、hgBA、plp B和oma87共7种毒力基因,A2型溶血性曼氏杆菌分离株携带irp、ompA、plpE、tbpA、tbpB、fhaC、lkt、lpsA、nanH和gcp共10种毒力基因。2株分离菌对阿卡米星、妥布霉素、红霉素和诺氟沙星均高度敏感,且均能导致小鼠死亡。结果表明,导致甘肃省某动物园盘羊死亡的病原菌可能为携带多种毒力基因的D型多杀性巴氏杆菌和A2型溶血性曼氏杆菌。本结果为该动物园盘羊呼吸道疾病的预防提供了参考,并为发病动物提供了适合的治疗方案。

To investigate and characterize the pathogenic bacteria responsible for the death of argali in a zoo in Gansu Province,the lung tissue was collected from one argali dying of respiratory disorders with cough,dyspnea and other clinical symptoms.Bacteria were isolated and cultured from the collected samples and identified by morphological observation as well as 16S rRNA gene sequencing.The isolated strains were further characterized by capsular serotype identification,virulence gene de-tection,drug sensitivity test and pathogenicity test in mice.The results showed that a highly pathogenic Pasteurella multocida strain of serotype D and Mannerella hemolytica strain of serotype A2 was isolated and identified from the lung tissue of the argali.The isolated strain of Pasteurella multocida serotype D carries 7 virulence genes including exbB,exbD,fur,tonB,hgBA,plpB and oma87,while the isolated strain Mannerella hemolytica serotype A2 carries 10 virulence genes including irp,ompA,plpE,tbpA,tbpB,fhaC,lkt,lpsA,nanH and gcp.The two isolates exhibited high susceptibility to amikacin,tobramycin,erythromycin,and norfloxacin.In addition,both of the two isolates were fatal to mice when inoculated.These results suggested that Pasteurella multocida serotype D and Mannerella hemolytica serotype A2 are the probable pathogens responsible for the death of the zoo argali in Gansu.The results also provided basis to carrying out measurements for prevention and treatment of the bacteria caused respiratory diseases in argali in the zoo.