| Congresso Brasileiro de Microbiologia 2023 | Resumo: 378-2 | ||||
Resumo:The frequency of carbapenem resistance rate has increased over the years. In aquatic ecosystems, the blaKPC gene has been associated with different clones of Klebsiella species, including the international high-risk clones of Klebsiella pneumoniae. In this regard, water samples are important carriers of potentially pathogenic strains and carbapenemase-encoding genes. In this study, we characterized carbapenem-resistant Klebsiella sp. isolates recovered from different Brazilian aquatic ecosystems. In 2021, 86 water samples from 58 cities were collected from rivers, streams, and lakes. The samples were filtered by the membrane filtration technique and the membranes were placed on plates of MacConkey agar supplemented with 4 mg/L of meropenem. Then, random colonies were selected and stored. Molecular identification was performed by conventional polymerase chain reactions (PCRs) using species-specific targets. In this sense, 12 K. pneumoniae and five Klebsiella quasipneumoniae were obtained. Disk diffusion, broth microdilution, and E-test® methods were performed to determine the antimicrobial susceptibility. The isolates were resistant to carbapenems, aminoglycosides, fluoroquinolones, folate pathway antagonists, tetracyclines, phenicols, polymyxins, and nitrofurans, being classified as multidrug- or extensively drug-resistant. The antimicrobial resistance genes (ARGs), virulence genes, plasmid replicons, and multilocus sequence typing (MLST) were determined by PCRs followed by Sanger sequencing. Several ARGs were observed, spotlighting the presence of blaKPC gene in all isolates, as well as the detection of blaGES and blaCTX-M in some isolates. Furthermore, the virulence genes entB, mrkD, ybtS, and iutA were found, while the most prevalent plasmid families were IncFIB and IncN. MLST analysis revealed a great diversity of sequence types (ST), highlighting the international high-risk clones (i.e., ST11 and ST340). A K. pneumoniae isolate, named EW1538, was selected for whole-genome sequencing using the Illumina MiSeq platform. Genomic analysis showed that EW1538 isolate belonged to ST11 O1/O2v1:KL64 and harbored the blaKPC-2 gene associated with Tn4401i isoform on an IncN-pST15 plasmid. Therefore, this study highlights the distribution of blaKPC-harboring Klebsiella species in water sources, alerting for the spread of multidrug resistance in the environment. Palavras-chave: Brazil, International high-risk clones, Multidrug resistance, water samples Agência de fomento:FAPESP (2021/01655-7), CAPES (88887.519091/2020-00, 8887.824722/2023-00, and Finance Code 001), and CNPq (304905/2022-4, 308914/2019-8, 130086/2021-5, 141016/2021-3, and 150712/2022-7) | |||||