Prevalence and Genetic Relatedness of Extended Spectrum-β-Lactamase-Producing Escherichia coli Among Humans, Cattle, and Poultry in Pakistan

Microb Drug Resist. 2019 Nov;25(9):1374-1381. doi: 10.1089/mdr.2018.0450. Epub 2019 Jul 3.

Abstract

Objective: To determine the prevalence and genetic relatedness of blaCTX-M-type extended spectrum-β-lactamase (ESBL)-producing Escherichia coli at the human-animal interface in Pakistan. Materials and Methods: A total of 150 human, cattle, and poultry fecal samples (50 each) were screened for ESBL-producing E. coli using ESBL CHROMagar®. Bacterial species confirmation as well as determination of minimum inhibitory concentrations (μg/mL) to different antibiotics was performed using the automated VITEK®-2 compact system. Phenotypic confirmation of ESBL production was performed according to the Clinical Laboratory Standards Institute (CLSI) guidelines. Genetic analysis of blaCTX-M was carried out by PCR and DNA sequencing. Plasmids and clonal similarity of the E. coli strains were determined by PCR-based replicon typing and pulsed-field gel electrophoresis (PFGE), respectively. Results: Of 150 samples, 29 (19.3%) ESBL-producing E. coli were recovered, and majority of them originated from human (n = 16; 55%), followed by cattle (n = 9; 31%) and poultry (n = 4; 13.7%). blaCTX-M-15 was predominant ESBL genotype (n = 25; 86.2%), mainly identified from human (n = 15) and cattle (n = 9). This is also the first report of the occurrence of CTX-M-15 and CTX-M-55 in cattle and poultry E. coli isolates of Pakistan, respectively. The majority of the ESBL-producing E. coli (96.5%) showed a multidrug resistance phenotype. All isolates carried IncFII or IncFIA plasmids, and the phylogroup B1 was dominant (44.8%) followed by phylogroups A (31%), D (17.2%), and B2 (6.8%). PFGE revealed that isolates from different hosts were genetically unrelated. Conclusion: Presence of CTX-M-15-type ESBL-producing E. coli in different reservoirs is alarming and has the potential to impact both veterinary and human therapeutic treatment options.

Keywords: Escherichia coli; MIC; blaCTX-M; genetic relatedness; replicon typing.

MeSH terms

  • Adult
  • Animals
  • Anti-Bacterial Agents / pharmacology*
  • Cattle / microbiology
  • Electrophoresis, Gel, Pulsed-Field
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • Escherichia coli / isolation & purification*
  • Escherichia coli Infections / epidemiology*
  • Escherichia coli Infections / microbiology
  • Escherichia coli Infections / veterinary
  • Escherichia coli Proteins / genetics
  • Humans
  • Microbial Sensitivity Tests
  • Middle Aged
  • Pakistan / epidemiology
  • Plasmids / genetics
  • Poultry / microbiology
  • Prevalence
  • beta-Lactamases / genetics*

Substances

  • Anti-Bacterial Agents
  • Escherichia coli Proteins
  • beta-lactamase CTX-M-15
  • beta-Lactamases
  • beta-lactamase CTX-M-55, E coli