野生鸟类分离的多重耐药肺炎克雷伯氏菌(Klebsiella pneumoniae)耐药基因分子进化特征研究  

Molecular evolutionary characterization of multidrug-resistant Klebsiella pneumoniae genes isolated from wild birds

作　　者：王雪 王猛 赵佳男 季芳 武斌 王学静[3] 秦建华[2] 王承民 WANG Xue;WANG Meng;ZHAO Jia՚nan;JI Fang;WU Bin;WANG Xuejing;QIN Jianhua;WANG Chengmin(Guangdong Key Laboratory of Animal Conservation and Resource Utilization,Guangdong Public Laboratory of Wild Animal Conservation and Utilization,Institute of Zoology,Guangdong Academy of Sciences,Guangzhou 510260,Guangdong,China;College of Veterinary Medicine,Hebei Agricultural University,Baoding 071000,Hebei,China;Hebei Institute of Animal Husbandry and Veterinary Medicine,Baoding 071000,Hebei,China)

机构地区：[1]广东省科学院动物研究所,广东省动物保护与资源利用重点实验室,广东省野生动物保护与利用公共实验室,广东广州510260 [2]河北农业大学动物医学院,河北保定071000 [3]河北省畜牧兽医研究所,河北保定071000

出　　处：《微生物学报》2022年第11期4477-4493,共17页Acta Microbiologica Sinica

基　　金：广东省科学院人才专项(2016GDASRC-0205);北京动物园北京市重点实验室开放课题(ZDK202105)。

摘　　要：【目的】调查野生鸟类携带菌的耐药状况,探索其在细菌耐药性传播过程中的作用。【方法】从野生鸟类石鸡、绯胸鹦鹉、太阳锥尾鹦鹉和黑领椋鸟的新鲜粪便分离4株Klebsiella pneumoniae,采用微量肉汤稀释法评估其多重耐药表型,并利用全基因组测序技术和细菌全因组关联分析、比较基因组学方法对分离株进行分子溯源,系统解析其携带的多重耐药质粒或基因与其宿主、同源质粒间的关联。【结果】4株肺炎克雷伯菌的耐药谱各不相同,来自石鸡样本的分离株S90-2对9种药物耐受,绯胸鹦鹉样本分离株S141对3种药物耐受,太阳锥尾鹦鹉分离株M911-1仅耐受氨苄西林,黑领椋鸟的样本分离株S130-1对所使用的14种药物完全敏感。S90-2属于ST629型,携带bla、fosA6、aac(3)-Iid和bla为主的30个耐药基因和携带1个耐药性质粒pS90-2.3(IncR型)。S141属于ST1662型,携带fosA5、bla等27个耐药基因,1个质粒pS141.1[IncFIB(K)(p CAV1099-114)/rep B型]仅携带耐药基因ade F。M911-1为新ST类型,携带blaSHV-1、fosA6等共计27个耐药基因,其质粒pM911-1.1携带了3个耐药基因。S130-1属于ST3753型,携带bla、fosA6等27个耐药基因,pS130-1[IncFIB(K)型]则仅携带一个耐药基因tet(A)。质粒比对表明,质粒pS90-2.3携带的耐药基因片段源自不同的肠杆菌科菌株染色体或质粒。pS90-2.3的同源质粒主要来自人类宿主菌,且主要在中国分布,这些质粒主要细菌宿主为K. pneumoniae和Escherichia coli,且ST11型K. pneumoniae分离株为重要宿主菌。【结论】本研究中来自野生鸟类的多重耐药K.pneumoniae,其耐药基因主要来自质粒,质粒耐药基因主要由转座子、插入序列、整合子和前噬菌体等可移动元件介导,这些多重耐药质粒与人类的宿主菌密切相关。[Objective]To investigate the drug resistance status of wild birds carrying bacteria and to explore their role in the transmission of bacterial drug resistance. [Methods]Four Klebsiella pneumoniae were isolated from fresh feces of captured Alectoris chukar, Psittacula alexandri, Aratinga solstitialis and Sturnus nigricollis, and were assessed for multidrug resistance phenotypes by micro-broth dilution method. Bacterial genome-wide association analysis and comparative genomics were used to trace the isolates and systematically analyze the association between the multidrug resistance plasmids/genes and their hosts/homologous plasmids. [Results]Four strains of K. pneumoniae showed different drug resistance phenotypes. Specifically, S90-2 from A. chukar was resistant to nine drugs including ampicillin, cefuroxime, cefazolin, ceftriaxone and cefepime;S141 from P. alexandri was resistant to ampicillin, cefuroxime and cefazolin;M911-1 from A. solstitialis was resistant to ampicillin only;S130-1 from S. nigricollis was sensitive to all of the 14 drugs. S90-2 belonged to ST629 type and carried 30 resistance genes including bla, fosA6, aac(3)-Iid and bla, and its plasmid pS90-2.3(IncR) carried resistance genes of mph A, dfrA12, aadA2, qacEdelta1, sul1, tet(A), aph(3’)-Ia, sul2 and aph(3’’)-Ib. S141 belonged to ST1662 type and carried 27 resistance genes including fosA5 and bla, and only plasmid pS141.1 [IncFIB(K)(pCAV1099-114)/repB]carried one resistance gene ade F. M911-1 was a new ST type, carrying 27 resistance genes such as blaSHV-1 and fosA6, and its plasmid pM911-1.1(novel) carried three resistance genes qnrS1, blaLAP-2 and tet(A). S130-1 belonged to ST3753 type, carrying 27 resistance genes such as blaSHV-11 and fosA6, and its plasmid pS130-1[IncFIB(K)]carried only one resistance gene tet(A). The plasmids pM911-1.1 and pS90-2.3 failed to perform conjugative transfer, but their resistance gene fragments were derived from multiple homologous chromosomes or plasmids of Enterobacteriaceae strains, and the formatio

关 键 词：野生鸟类 多重耐药菌 肺炎克雷伯氏菌 耐药质粒 

分 类 号：S852.61[农业科学—基础兽医学]