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意大利伴侣动物中克雷伯菌属的超广谱β-内酰胺酶、AmpCβ-内酰胺酶及质粒介导的喹诺酮耐药性

Extended-spectrum-beta-lactamases, AmpC beta-lactamases and plasmid mediated quinolone resistance in klebsiella spp. from companion animals in Italy.

作者信息

Donati Valentina, Feltrin Fabiola, Hendriksen Rene S, Svendsen Christina Aaby, Cordaro Gessica, García-Fernández Aurora, Lorenzetti Serena, Lorenzetti Raniero, Battisti Antonio, Franco Alessia

机构信息

Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana, Rome, Italy.

Technical University of Denmark, National Food Institute (DTU-Food), Kongens Lyngby, Denmark.

出版信息

PLoS One. 2014 Mar 4;9(3):e90564. doi: 10.1371/journal.pone.0090564. eCollection 2014.

Abstract

We report the genetic characterization of 15 Klebsiella pneumoniae (KP) and 4 isolates of K. oxytoca (KO) from clinical cases in dogs and cats and showing extended-spectrum cephalosporin (ESC) resistance. Extended spectrum beta-lactamase (ESBL) and AmpC genes, plasmid-mediated quinolone resistance (PMQR) and co-resistances were investigated. Among KP isolates, ST101 clone was predominant (8/15, 53%), followed by ST15 (4/15, 27%). ST11 and ST340, belonging to Clonal Complex (CC)11, were detected in 2012 (3/15, 20%). MLST on KP isolates corresponded well with PFGE results, with 11 different PFGE patterns observed, including two clusters of two (ST340) and four (ST101) indistinguishable isolates, respectively. All isolates harbored at least one ESBL or AmpC gene, all carried on transferable plasmids (IncR, IncFII, IncI1, IncN), and 16/19 were positive for PMQR genes (qnr family or aac(6')-Ib-cr). The most frequent ESBL was CTX-M-15 (11/19, 58%), detected in all KP ST101, in one KP ST15 and in both KP ST340. blaCTX-M-15 was carried on IncR plasmids in all but one KP isolate. All KP ST15 isolates harbored different ESC resistance genes and different plasmids, and presented the non-transferable blaSHV-28 gene, in association with blaCTX-M-15, blaCTX-M-1 (on IncR, or on IncN), blaSHV-2a (on IncR) or blaCMY-2 genes (on IncI1). KO isolates were positive for blaCTX-M-9 gene (on IncHI2), or for the blaSHV-12 and blaDHA-1 genes (on IncL/M). They were all positive for qnr genes, and one also for the aac(6')-Ib-cr gene. All Klebsiella isolates showed multiresistance towards aminoglycosides, sulfonamides, tetracyclines, trimethoprim and amphenicols, mediated by strA/B, aadA2, aadB, ant (2")-Ia, aac(6')-Ib, sul, tet, dfr and cat genes in various combinations. The emergence in pets of multidrug-resistant Klebsiella with ESBL, AmpC and PMQR determinants, poses further and serious challenges in companion animal therapy and raise concerns for possible bi-directional transmission between pets and humans, especially at household level.

摘要

我们报告了从犬猫临床病例中分离出的15株肺炎克雷伯菌(KP)和4株产酸克雷伯菌(KO)的基因特征,这些菌株表现出对超广谱头孢菌素(ESC)耐药。对超广谱β-内酰胺酶(ESBL)和AmpC基因、质粒介导的喹诺酮耐药性(PMQR)及共耐药性进行了研究。在KP分离株中,ST101克隆占主导地位(8/15,53%),其次是ST15(4/15,27%)。属于克隆复合体(CC)11的ST11和ST340在2012年被检测到(3/15,20%)。对KP分离株进行的多位点序列分型(MLST)结果与脉冲场凝胶电泳(PFGE)结果吻合良好,共观察到11种不同的PFGE模式,包括分别由两个(ST340)和四个(ST101)难以区分的分离株组成的两个聚类。所有分离株均携带至少一个ESBL或AmpC基因,且均携带于可转移质粒(IncR、IncFII、IncI1、IncN)上,16/19株对PMQR基因(qnr家族或aac(6')-Ib-cr)呈阳性。最常见的ESBL是CTX-M-15(11/19,58%),在所有KP ST101、一株KP ST15和两株KP ST340中均有检测到。除一株KP分离株外,blaCTX-M-15均携带于IncR质粒上。所有KP ST15分离株均携带不同的ESC耐药基因和不同质粒,并呈现与blaCTX-M-15、blaCTX-M-1(位于IncR或IncN上)、blaSHV-2a(位于IncR上)或blaCMY-2基因(位于IncI1上)相关的不可转移的blaSHV-28基因。KO分离株对blaCTX-M-9基因(位于IncHI2上)或blaSHV-12和blaDHA-1基因(位于IncL/M上)呈阳性。它们对qnr基因均呈阳性,一株还对aac(6')-Ib-cr基因呈阳性。所有克雷伯菌分离株均对氨基糖苷类、磺胺类、四环素类、甲氧苄啶和氯霉素表现出多重耐药性,由strA/B、aadA2、aadB、ant(2")-Ia、aac(6')-Ib、sul、tet、dfr和cat基因以各种组合介导。携带ESBL、AmpC和PMQR决定簇的多重耐药克雷伯菌在宠物中的出现,给伴侣动物治疗带来了进一步的严峻挑战,并引发了对宠物与人类之间可能双向传播的担忧,尤其是在家庭层面。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/403e/3942433/20149d9f31f1/pone.0090564.g001.jpg

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