Analysis of extended-spectrum-β-lactamase-producing Escherichia coli isolates collected in the GERM-Vet monitoring programme

J Antimicrob Chemother. 2013 Aug;68(8):1741-9. doi: 10.1093/jac/dkt123. Epub 2013 Apr 18.

Abstract

Objectives: The aims of this study were (i) to detect extended-spectrum β-lactamase (ESBL) genes among 1378 Escherichia coli isolates from defined disease conditions of companion and farm animals and (ii) to determine the localization and organization of ESBL genes.

Methods: E. coli isolates from the German resistance monitoring programme GERM-Vet were included in the study. Plasmids were transferred by conjugation or transformation and typed by PCR-based replicon typing. ESBL genes were detected by PCR; the complete ESBL genes and their flanking regions were sequenced by primer walking. Phylogenetic grouping and multilocus sequence typing (MLST) were performed for all ESBL-producing E. coli isolates.

Results: Of the 27 ESBL-producing E. coli isolates detected, 22 carried blaCTX-M-1 genes on IncN (n = 16), IncF (n = 3), IncI1 (n = 2) or multireplicon (n = 1) plasmids. A blaCTX-M-3 gene was located on an IncN plasmid and a blaCTX-M-15 gene was located on an IncF plasmid. A multireplicon plasmid and an IncHI1 plasmid harboured blaCTX-M-2. A blaTEM-52c gene was identified within Tn2 on an IncI1 plasmid. The blaCTX-M genes located within the same or related genetic contexts showed differences due to the integration of insertion sequences. Various MLST types were detected, with ST10 (n = 7), ST167 (n = 4) and ST100 (n = 3) being the most common.

Conclusions: This study showed that the blaCTX-M-1 gene is the predominant ESBL gene among E. coli isolates from diseased animals in Germany and a considerable structural heterogeneity was found in the regions flanking the blaCTX-M-1 gene. Insertion sequences, transposons and recombination events are likely to be involved in alterations of the ESBL gene regions.

Keywords: ESBLs; MLST; diseased animals; plasmids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Domestic
  • Cluster Analysis
  • Conjugation, Genetic
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Escherichia coli / enzymology*
  • Escherichia coli / isolation & purification
  • Escherichia coli Infections / microbiology
  • Escherichia coli Infections / veterinary*
  • Gene Transfer, Horizontal
  • Germany
  • Molecular Sequence Data
  • Multilocus Sequence Typing
  • Pets
  • Phylogeny
  • Plasmids
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sequence Homology
  • Transformation, Bacterial
  • beta-Lactamases / genetics*

Substances

  • DNA, Bacterial
  • beta-Lactamases

Associated data

  • GENBANK/HF549088
  • GENBANK/HF549089
  • GENBANK/HF549090
  • GENBANK/HF549091
  • GENBANK/HF549092
  • GENBANK/HF549093
  • GENBANK/HF549094
  • GENBANK/HF549095