Characterization of a blaNDM-1-carrying IncHI5 plasmid from Enterobacter cloacae complex of food-producing animal origin

Abstract

Objectives To characterize an NDM-1-encoding multiresistance IncHI5 plasmid from Enterobacter cloacae complex of chicken origin. Methods Carbapenemase genes were detected by PCR and Sanger sequencing. The MICs for the E. cloacae complex isolate and its transformant were determined by the agar dilution and broth microdilution methods. Conjugation and electrotransformation were performed to assess the horizontal transferability of the carbapenemase plasmid. Plasmid DNA was isolated from the transformant and fully sequenced using Illumina HiSeq and PacBio platforms. Plasmid stability was investigated by sequential passages on antibiotic-free medium. A circular intermediate was detected by inverse PCR and Sanger sequencing. Results Plasmid pNDM-1-EC12 carried a conserved IncHI5 backbone and exhibited an MDR phenotype. All antimicrobial resistance genes were clustered in a single MDR region. Genetic environment analysis revealed that the blaNDM-1 gene was in a novel complex integron, In469. Based on sequence analysis, the blaNDM-1-carrying region was thought to be inserted by homologous recombination. Inverse PCR indicated that an ISCR1-mediated circular intermediate can be formed. Plasmid pNDM-1-EC12 was stably maintained both in the parental strain and the transformant without selective pressure. Comprehensive analysis of IncHI5-type plasmids suggested that they may become another key vehicle for rapid transmission of carbapenemase genes. Conclusions To the best of our knowledge, this is the first report of a fully sequenced IncHI5 plasmid recovered from an E. cloacae complex strain of food-producing animal origin. Co-occurrence of blaNDM-1 with genes encoding resistance to other antimicrobial agents on the same IncHI5 plasmid may result in the co-selection of blaNDM-1 and facilitates its persistence and rapid dissemination.

Wide Distribution and Specific Resistance Pattern to Third-Generation Cephalosporins of Enterobacter cloacae Complex Members in Humans and in the Environment in Guadeloupe (French West Indies)

“…In accordance with our observations, they have rarely been observed in samples of other origins, except from companion and wild animals (Haenni et al, 2016;Harada et al, 2017;Goldberg et al, 2019). Different sample types shared a few lineages, one being ST90 (n = 5), which is also involved in human and animals infections (Peirano et al, 2018;Zhu et al, 2020). Moreover, two ST90 strains isolated from an anole and a raw water sample harbored genes encoding for the yersiniabactin system.…”

Characterization of a blaNDM-1-Bearing IncHI5-Like Plasmid From Klebsiella pneumoniae of Infant Origin

“…Online BLAST against the NCBI nr database revealed that it shared an overall similar backbone with plasmid pNDM-1-EC12 (MN598004), pKP-13-14-NDM-9 (MN175386), p104922-NDM (MT062912), pKP19-3023-374kb (CP063748) and pKP19-3088-375kb (CP063149), including genes essential for replication (repHI5B and repFIB-like), partition (parAB) and conjugal transfer (tra1 and tra2) (Figure 1). As previously reported, pNDM-1-EC12 was identified as an IncHI5 plasmid (Zhu et al, 2020), whereas pKP19-3023-374kb and pKP19-3088-375kb belonged to IncHI5-like plasmids (Qin et al, 2021). The replication genes of IncHI5-like plasmids shared 97.06% (repHI5B) and 97.99% (repFIB-like) nucleotide identity with those of IncHI5 plasmids, while the repHI5B protein and repFIB-like protein of IncHI5-like plasmids shared 97.97% and 97.27% amino acid identity with those of IncHI5 plasmids.…”

“…Members of the IncHI group contained five types of plasmids (Liang et al, 2018), of which considerable genetic conservation was detected within subgroups, but the conserved sequences of each subgroup were dramatically distinguished from each other (Liang et al, 2017). IncHI5 plasmids are associated with various carbapenemase genes (Zhu et al, 2020), implying that investigations are needed to continuously monitor the prevalence of IncHI5 plasmids in different sources, especially among clinical settings. More seriously, a recent study reported two IncHI5-like plasmids co-harboring carbapenem resistance genes and tigecycline resistance module tmexCD1-toprJ1 (Qin et al, 2021), indicating that the IncHI5 plasmid has evolved and may develop as key vectors to carry important and novel antimicrobial resistance determinants.…”

Dietary Factors of blaNDM Carriage in Health Community Population: A Cross-Sectional Study

“…The emergence and spread of CRE carrier and infections are serious threats to public health worldwide. Carbapenem-resistant gene bla NDM was frequently found from non-human sources [8][9][10][11][12][13][14][15]. There is an ongoing debate as to what extent antimicrobial resistance (AMR) can be transmitted from animals to humans via the consumption of animal products.…”

“…While a growing number of carbapenemases found from non-human sources have been reported [4][5][6][7]. Despite the expansion of the New Delhi metallo-β-lactamase-1 (NDM-1) worldwide, recent years NDM has been frequently found from non-human sources such as food animals, vegetables and water in China [8][9][10][11][12][13][14][15]. Since the food chain has serve as a reservoir for resistance genes, due to the lack of epidemiological data on the role of diet in resistance genes in humans, there is an ongoing debate as to what extent AMR such as carbapenem resistance can be transmitted from dietary intake to humans via the consumption of food products [16].…”