Abstract
ObjectiveTo investigate the molecular characteristics of extended-spectrum cephalosporin (ESC)-resistant Enterobacteriaceae collected during a cross-sectional study examining the prevalence and risk factors for faecal carriage of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae in humans living in areas with high or low broiler density.MethodsESC-resistant Enterobacteriaceae were identified by combination disc-diffusion test. ESBL/AmpC/carbapenemase genes were analysed using PCR and sequencing. For E. coli, phylogenetic groups and MLST were determined. Plasmids were characterized by transformation and PCR-based replicon typing. Subtyping of plasmids was done by plasmid multilocus sequence typing.Results175 ESC-resistant Enterobacteriaceae were cultured from 165/1,033 individuals. The isolates were Escherichia coli(n=65), Citrobacter freundii (n=52), Enterobacter cloacae (n=38), Morganella morganii (n=5), Enterobacter aerogenes (n=4), Klebsiella pneumoniae (n=3), Hafnia alvei (n=2), Shigella spp. (n=2), Citrobacter amalonaticus (n=1), Escherichia hermannii (n=1), Kluyvera cryocrescens (n=1), and Pantoea agglomerans (n=1). The following ESBL genes were recovered in 55 isolates originating from 49 of 1,033 (4.7 %) persons: bla CTX-M-1 (n=17), bla CTX-M-15 (n=16), bla CTX-M-14 (n=9), bla CTX-M-2 (n=3), bla CTX-M-3 (n=2), bla CTX-M-24 (n=2), bla CTX-M-27 (n=1), bla CTX-M-32 (n=1), bla SHV-12 (n=2), bla SHV-65 (n=1) and bla TEM-52 (n=1). Plasmidic AmpC (pAmpC) genes were discovered in 6 out of 1,033 (0.6 %) persons. One person carried two different E. coli isolates, one with bla CTX-M-1 and the other with bla CMY-2 and therefore the prevalence of persons carrying Enterobacteriaceae harboring ESBL and/or pAmpC genes was 5.2 %. In eight E. coli isolates the AmpC phenotype was caused by mutations in the AmpC promoter region. No carbapenemase genes were identified. A large variety of E. coli genotypes was found, ST131 and ST10 being most common.ConclusionsESBL/pAmpC genes resembled those from patients in Dutch hospitals, indicating that healthy humans form a reservoir for transmission of these determinants to vulnerable people. The role of poultry in the transmission to humans in the community remains to be elucidated.
Highlights
Extended-spectrum-β-lactamase/AmpC producing Enterobacteriaceae have been found among humans worldwide
175 extended spectrum cephalosporins (ESC)-resistant Enterobacteriaceae were cultured from 165/1,033 individuals
The following ESBL genes were recovered in 55 isolates originating from 49 of 1,033 (4.7 %) persons: blaCTX-M-1 (n=17), blaCTX-M-15 (n=16), blaCTX-M-14 (n=9), blaCTX-M-2 (n=3), blaCTX-M-3 (n=2), blaCTX-M-24 (n=2), blaCTX-M-27 (n=1), blaCTX-M-32 (n=1), blaSHV-12 (n=2), blaSHV-65 (n=1) and blaTEM-52 (n=1)
Summary
Extended-spectrum-β-lactamase/AmpC producing Enterobacteriaceae have been found among humans worldwide. Most large-scale studies in humans, report data of patients or travelers and/or focus on ESBL-producing bacteria and/or certain bacterial species only (e.g. Escherichia coli or Klebsiella pneumoniae) [1,2,3,4]. Data on the prevalence of fecal carriage of ESBL/AmpC/carbapenemase producing Enterobacteriaceae in healthy humans in the community are scarce. In 2011, a cross-sectional study was performed to determine the prevalence of, and identify risk factors for, carriage of ESBL-producing Enterobacteriaceae in people living in municipalities with either high or low broiler densities [14]. The prevalence of carriage of ESBL-producing bacteria was 5.1% and this percentage was lower in municipalities with high broiler densities (3.6%) compared to municipalities with low broiler densities (6.7%) [14]. The aim of the present study was to analyse the isolates from this cross-sectional study, including isolates with an AmpC phenotype, with respect to molecular characteristics and compare them to published data on isolates from patients, broilers, and persons in contact with broilers
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