Characterization of NDM-5 Carbapenemase-Encoding Gene (blaNDM-5) – Positive Multidrug Resistant Commensal Escherichia coli from Diarrheal Patients

Abstract

Purpose The multidrug resistance Enterobacteriaceae cause many serious infections resulting in prolonged hospitalization, increased treatment charges and mortality rate. In this study, we characterized bla NDM-5 -positive multidrug resistance commensal Escherichia coli (CE) isolated from diarrheal patients in Kolkata, India. Methods Three CE strains were isolated from diarrheal stools, which were negative for different pathogroups of diarrheagenic E. coli (DEC). The presence of carbapenemases encoding genes and other antimicrobial resistance genes (ARGs) was detected using PCR. The genetic arrangement adjoining bla NDM-5 was investigated by plasmid genome sequencing. The genetic relatedness of the strains was determined by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) methods. Results In addition to colistin, the bla NDM-5 -positive CE strains showed resistance to most of the antibiotics. Higher MICs were detected for ciprofloxacin (>32 mg/L) and imipenem (8 mg/L). Molecular typing revealed that three CE strains belonged to two different STs (ST 101 and ST 648) but they were 95% similar in the PFGE analysis. Screening for ARGs revealed that CE strains harbored Int-1, bla TEM , bla C TX-M3 , bla OXA-1 , bla OXA-7 , bla OXA-9 , tetA, strA, aadA1, aadB, sul2, floR, mph(A) , and aac(6´)-Ib-cr . In conjugation experiment, transfer frequencies ranged from 2.5×10 −3 to 8.4x10 −5 . The bla NDM-5 gene was located on a 94-kb pNDM-TC-CE-89 type plasmid, which is highly similar to the IncFII plasmid harboring an IS26-IS30- bla NDM-5 - ble MBL - trpF-dsbd -IS 91-dhps structure. Conclusion To the best of our knowledge, this is the first report on carbapenem resistance involving the bla NDM-5 gene in CE from diarrheal patients. The circulation of bla NDM-5 gene in CE is worrisome, since it has the potential to transfer bla NDM-5 gene to other enteric pathogens.

Coexistence of blaNDM–5 and tet(X4) in international high-risk Escherichia coli clone ST648 of human origin in China

“…The ECCL209 isolate was assigned as serotype O83:H42 using SerotypeFinder 2.0, 5 which is an extraintestinal pathogenic E. coli (ExPEC) primarily found in samples from animals, indicating their possible transmission from animal to humans (Abreu-Salinas et al, 2020;Shafiq et al, 2021aShafiq et al, ,b, 2022. MLST analysis revealed that E. coli isolate ECCL209 in this study belonged to sequence type (ST648), which had been previously reported to carry bla CTX−M− , bla CMY−2− , bla NDM− , bla OXA−48− , and mcr-1 encoding genes and caused a significant proportion of infections in humans (Hornsey et al, 2011;Poirel et al, 2018;Chowdhury et al, 2022). This clonal lineage has emerged as a pandemic high-risk clone, being globally reported in humans, animals, and the environment (Hornsey et al, 2011;Fernandes et al, 2018;Furlan et al, 2020;Chowdhury et al, 2022;Landolsi et al, 2022).…”

“…MLST analysis revealed that E. coli isolate ECCL209 in this study belonged to sequence type (ST648), which had been previously reported to carry bla CTX−M− , bla CMY−2− , bla NDM− , bla OXA−48− , and mcr-1 encoding genes and caused a significant proportion of infections in humans (Hornsey et al, 2011;Poirel et al, 2018;Chowdhury et al, 2022). This clonal lineage has emerged as a pandemic high-risk clone, being globally reported in humans, animals, and the environment (Hornsey et al, 2011;Fernandes et al, 2018;Furlan et al, 2020;Chowdhury et al, 2022;Landolsi et al, 2022). To the best of our knowledge, this is the first report of ST648, with bla NDM and tet(X4).…”

Whole genome-based genetic insights of bla NDM producing clinical E. coli isolates in hospital settings of Pakistant

“…Conjugation experiments were carried out to determine the transferability of plasmid-mediated bla NDM genes using sodium azide-resistant E. coli J53 as the recipient strain, as previously described ( 28 ). Briefly, isolates were grown on chromogenic media plates with 1 µg/mL meropenem (AstraZeneca, London, UK) and E. coli J53 on chromogenic media with 200 µg/mL sodium azide.…”

Genomic Insights into the First Emergence of blaNDM-5-Carrying Carbapenem-Resistant Salmonella enterica Serovar London Strain in China

“…These evidence collectively suggest that the bla NDM-5 -carrying cassette is frequently transferred as an integral complex with class 1 integron rather than as a standalone gene cassette, as also implicated in previous studies with E. coli. 47,48 Nevertheless, future research is necessary to validate the role of class 1 integron and ISCR1 element in the dissemination of bla NDM-5 gene. Using cgSNP-based phylogenetic analysis, we found that A132 shares a close genomic relationship with numerous Chinese S. London ST155 strains retrieved from EnteroBase.…”