貉源肺炎克雷伯菌的分离鉴定及药敏试验

doi:10.16431/j.cnki.1671-7236.2019.04.030

摘要/Abstract

摘要：

为确定河北昌黎地区某养殖场貉呼吸道疾病引起死亡的病因，本研究从送检的貉病料肺脏中分离到1株革兰氏阴性短杆菌，并对分离株进行形态学观察、16S rRNA序列测序及全自动细菌生化鉴定仪鉴定，在此基础上完成了分离株的分型、人工感染小鼠试验及药敏试验等。结果显示，该分离株为非K1/K2型肺炎克雷伯菌，并将其命名为CLKp18；该分离株的16S rRNA基因序列与GenBank中登录的多株肺炎克雷伯菌参考株同源性均高达99%。Vitek全自动细菌鉴定仪结果表明，该菌株与肺炎克雷伯菌鉴定标准相似度为99%。人工感染小鼠试验结果显示，试验组小鼠在攻毒后6 h开始出现临床症状。无菌取出死亡小鼠的肺脏、肝脏、脾脏及肾脏，划线涂板，均可分离到该菌，表明该分离株感染小鼠后，能侵入到小鼠体内多个组织器官，引起不同程度的病变，尤以肝脏和肺脏病变较为明显，对小鼠的半数致死量（LD₅₀）为2.17×10⁷ CFU。药敏试验结果表明，该菌对单环内酰胺类、氯霉素类、氨基糖苷类、四环素类、磺胺类及大部分头孢类耐药，但对阿米卡星、阿莫西林/克拉维酸、头孢他啶和亚胺培南敏感。本研究结果确定了引起本次貉呼吸道症状疾病的致病菌为非K1/K2型肺炎克雷伯菌，且该分离株有较强的致病性和多重耐药性。本研究为临床治疗由该菌引起的貉肺炎提供了参考依据。

关键词: 貉; 肺炎克雷伯菌; 分离鉴定; 药敏试验

Abstract:

To determine the pathogen caused a serious mortality of racoon dog in Hebei province,and one Gram-negative bacterium was isolated from the the lung samples of diseased racoon dog.The isolate received morphological obsrvation,16S rRNA sequence analysis and biochemical test.Basis on this,the serotype characteristics,pathogenicity tests,drug sensitivity test were completed.The isolated strain was confirmed as not K1/K2 Klebsiella pneumoniae,named as CLKp18.16S rRNA gene sequence analysis results showed that the bacterium was 99% homologous with many strains of Klebsiella accessed in GenBank.The Vitek automatic bacterial identification results showed that the similarity between the strain and Klebsiella pneumoniae standard was 99%.Pathogenicity test results showed that the mice in the experimental group began to develop clinical symptoms 6 h after the challenge.The lung,liver,spleen and kidney of the dead mice were aseptically removed,and the bacteria were isolated by scribing and plating.It indicated that the infected strain could invade many tissues and organs in mice after infection,which caused different degrees of lesions,especially liver and lung lesions,the LD₅₀ in mice was 2.17×10⁷ CFU.Drug susceptibility test results showed that the strain resistant to monocyclic lactam,chloramphenicol,aminoglycosides,tetracyclines,sulfonamides and most cephalosporins,and sensitive to amikacin,amoxycillin/clavulanic acid,imipenem and ceftazidime.This study confirmed that K1/K2 Klebsiella pneumoniae didn't cause the pathogens,and the strain with strong pathogenicity in mice and multi-drug resistance.The results provided a reference for clinical treatment of pneumonia caused by the strain.

Key words: racoon dog; Klebsiella pneumonia; isolation and identification; drug susceptibility test

中图分类号:

S852.61⁺2

韩坤, 吕娇, 于莹, 王洋, 王雅文, 许皓, 陈立志, 冯二凯, 白雪, 闫喜军. 貉源肺炎克雷伯菌的分离鉴定及药敏试验[J]. 《中国畜牧兽医》, 2019, 46(4): 1209-1216.

HAN Kun, LYU Jiao, YU Ying, WANG Yang, WANG Yawen, XU Hao, CHEN Lizhi, FENG Erkai, BAI Xue, YAN Xijun. Isolation,Identification and Drug Susceptibility of Klebsiella pneumoniae Isolated from Racoon Dog[J]. , 2019, 46(4): 1209-1216.

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