Congresso Brasileiro de Microbiologia 2023

Congresso Brasileiro de Microbiologia 2023

Resumo: 569-2

569-2

blaNDM-1 plasmid replacement in Klebsiella pneumoniae during the SARS-CoV-2 pandemic period.

Autores:

Mayana Kieling Hernandez Berdichevski (UFRGS - UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL , HCPA - HOSPITAL DE CLINICAS DE PORTO ALEGRE) ; Rafaela Ramalho Guerra (HCPA - HOSPITAL DE CLINICAS DE PORTO ALEGRE) ; Fabiana Caroline Zempulski Volpato (HCPA - HOSPITAL DE CLINICAS DE PORTO ALEGRE) ; Camila Mörschbächer Wilhelm (HCPA - HOSPITAL DE CLINICAS DE PORTO ALEGRE) ; Dariane Castro Pereira (HCPA - HOSPITAL DE CLINICAS DE PORTO ALEGRE) ; Andreza Fransciso Martins (UFRGS - UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL , HCPA - HOSPITAL DE CLINICAS DE PORTO ALEGRE) ; Afonso Luis Barth (HCPA - HOSPITAL DE CLINICAS DE PORTO ALEGRE, UFRGS - UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL )

Resumo:

Carbapenem Resistant Enterobacterales (CRE) is a global health threat due to the lack of effective treatments, especially among CRE isolates harboring more than one carbapenemase gene, such as blaKPC-2 and blaNDM-1. The spread of these genes occur by plasmid and transposon and thus, it is important to evaluate these genetic mobile elements to better understand their dissemination. In this study, a total of 362 carbapenem-resistance Gram-negative bacilli isolates were obtained from 2016 to 2023 to a surveillance study previously by Maldi-TOF and disc-diffusion methods. To evaluate the genetic environment, 20 isolates of K. pneumoniae were randomly selected by year, pre (before 2020) and post pandemic period (after 2020) to whole genome sequencing (WGS) that was performed by MiSeq (Illumina) s and by MinIon (Oxford Nanopore). Contig assembling was performed in QIAGEN CLC Genomics Workbench and plasmid assembling was analyzed in Geneious Prime using Genbank and Bowtie2. The annotation was performed by Prokka and curated manually in Geneious using BLAST. Antimicrobial resistance genes were identified in silico using QMI-AR. PlasmidFinder was used for predicting incompatibility group. BRIG was used for circular comparisons among plasmids. Data from WGS evidenced that 70% (14/20) of the isolates belong to ST16, 25% (5/20) to ST11 and 5% (1/20) to ST307 and all isolates carried blaKPC-2 and blaNDM-1 by distinct plasmids which also carried other resistance genes. The blaNDM-1 gene was identified in 90% (18/20) of the isolates (all obtained after 2020) in a plasmid of 183,109 bp (pLB_10067) belonging to IncA/C group, added to genes that confers resistance to sulfonamides, aminoglycosides, macrolides, quinolones, amphenicols, tetracyclines and chlorhexidine. The blaNDM-1 was inserted between two IS26, an ISCR1 and an ISAba125 upstream of blaNDM-1, resulting in an IS26-flanked pseudo-composite transposon of 39,177bp. In isolates (2/20; 10%) recovered from 2016, the blaNDM-1 gene was carried by pLB_5236 (255,578 pb) plasmid that belongs to Inc group an IncHI1B/IncFIB, between two IS3000, in a Tn3000 of 11,824 bp that also harbors resistance genes to quinolones, aminoglycosides and macrolides. Our findings suggest a replacement of IncHI1B/IncFIB blaNDM-1 co-integrated plasmid by IncA/C during the pandemic period. It is known that smaller size plasmids, such as IncA/C have a higher capacity for horizontal transfer and lower fitness became its presence more favorable to bacteria. In conclusion, it is important to highlight that genomic surveillance programs are essential to improve the knowledge about the mobile genetic elements associated with antimicrobial resistance spread.

Palavras-chave:

co-integrated plasmids, blaKPC-2, blaNDM-1, K. pneumoniae co-harboring, spread of blaNDM-1

Agência de fomento:

Fundo de Incentivo a Pesquisa e Eventos do Hospital de Clínicas de Porto Alegre (FIPE/HCPA) and Instituto Nacional de Pesquisa em Resistência aos Antimicrobianos (INPRA/INCT).