| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1198-1 | ||||
Resumo:In 2019, the CDC listed Carbapenem-resistant Enterobacterales (CRE) as one of the most urgent threats to public health. Resistance to carbapenems is mainly due to the production of carbapenemases, and carbapenemase genes, such as blaNDM-1, are often harbored by mobile genetic elements that can promote the rapid spread of resistance. In this study, we reported the spread of an InA/C plasmid harbored blaNDM-1 among different bacterial species into a hospital in southern Brazil. E. coli was isolated from a blood sample collected in November 2022 from a patient hospitalized at Hospital de Clínicas de Porto Alegre (HCPA). The species identification was performed using VITEK®MALDI-TOF MS (bioMérieux, France) and the antimicrobial susceptibility profile was determined by disk diffusion according to BRCAST guidelines. Genomic DNA was extracted using a QIAamp DNA Mini Extraction Kit (QIAGEN®) and the concentration and quality were determined by Qubit (Thermo Fisher Scientific) and NanoDrop™ respectively. The paired-end library was constructed with the Nextera XT DNA Library Prep Kit (Illumina) to sequence the whole genome by Illumina MiSeq (2x250 bp paired-end reads). The raw short reads were assembled using CLC Genomic Workbench 23 and antimicrobial resistance genes were identified in silico using the QIAGEN Microbial Insight-Antimicrobial Resistance database (QMI-AR). PubMLST, ISFinder, PlasmidFinder and MobileElementFinder were used to characterize the ST and mobile genetic elements. Comparative genomic analysis was performed using Geneious Prime 9.0.5 and BRIG. E. coli isolate was classified as multidrug-resistant due a phenotype of resistance to all beta-lactams and also quinolones, sulfonamides, and tetracyclines. The draft genome consists of 4,790,398 bp, and de novo assembly resulted in 128 contigs (N50 120,012 bp; G+C content 50.6%). E. coli belongs to ST744, a high-risk international clone, especially associated with resistance to carbapenems and polymyxins. The erm(B), aadA5, aph(3’)-VI, aph(6)-lb, aph(3’’)-lb, aph(3’)-la, aac(6’)-lb-cr, tet(A), tet(B), sul, dfrA17, mph(A), catB3, flor, ARR-3, blaOXA-1, blaTEM-1B and blaNDM-1 resistance genes were identified encoding resistance to macrolides, aminoglycosides, tetracyclines, sulfonamides, amphenicols, quinolones, and beta-lactams. In particular, blaNDM-1 encodes resistance to carbapenems which are the main antimicrobials used in the treatment of serious infections caused by gram-negative bacteria. The blaNDM-1 gene was carried in an IncA/C plasmid-type that showed 98.8% similarity with a plasmid from the same incompatibility group, initially identified in Klebsiella pneumoniae (pLB_10067) isolated in 2020 at the same institution, evidencing the plasmid spread among different species and also suggesting its persistence in the institution over time. The blaNDM-1 was embedded in the classic genetic context of the IS26 flanked pseudo composite transposon containing ISCR1, usually associated with the mobilization of this gene. In addition, IncA/C plasmid co-harbored blaTEM-1B, blaOXA-1 and aac(6’)-lb-cr resistance genes that are spreading together with blaNDM-1. These findings highlight the hospital environment as a reservoir of mobile genetic elements that may be contributing to the intra- and inter-species spread of carbapenem resistance by horizontal gene transfer into the institution. Palavras-chave: blaNDM-1, Escherichia coli, IncA/C plasmid, inter-species spread Agência de fomento:CAPES, CNPQ | |||||