Diversity of Multi-Drug Resistant Avian Pathogenic Escherichia coli (APEC) Causing Outbreaks of Colibacillosis in Broilers during 2012 in Spain

PLoS One. 2015 Nov 23;10(11):e0143191. doi: 10.1371/journal.pone.0143191. eCollection 2015.

Abstract

Avian pathogenic Escherichia coli (APEC) are the major cause of colibacillosis in poultry production. In this study, a total of 22 E. coli isolated from colibacillosis field cases and 10 avian faecal E. coli (AFEC) were analysed. All strains were characterised phenotypically by susceptibility testing and molecular typing methods such as pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). The presence of 29 virulence genes associated to APEC and human extraintestinal pathogenic E. coli (ExPEC) was also evaluated. For cephalosporin resistant isolates, cephalosporin resistance genes, plasmid location and replicon typing was assessed. Avian isolates belonged to 26 O:H serotypes and 24 sequence types. Out of 22 APEC isolates, 91% contained the virulence genes predictors of APEC; iutA, hlyF, iss, iroN and ompT. Of all strains, 34% were considered ExPEC. PFGE analysis demonstrated a high degree of genetic polymorphism. All strains were multi-resistant, including those isolated from healthy animals. Eleven strains were resistant to cephalosporins; six contained blaCTX-M-14, two blaSHV-12, two blaCMY-2 and one blaSHV-2. Two strains harboured qnrA, and two qnrA together with aac(6')-Ib-cr. Additionally, the emergent clone O25b:H4-B2-ST131 was isolated from a healthy animal which harboured blaCMY-2 and qnrS genes. Cephalosporin resistant genes were mainly associated to the presence of IncK replicons. This study demonstrates a very diverse population of multi-drug resistant E. coli containing a high number of virulent genes. The E. coli population among broilers is a reservoir of resistance and virulence-associated genes that could be transmitted into the community through the food chain. More epidemiological studies are necessary to identify clonal groups and resistance mechanisms with potential relevance to public health.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biodiversity*
  • Chickens / microbiology*
  • Disease Outbreaks / statistics & numerical data*
  • Drug Resistance, Multiple, Bacterial* / genetics
  • Electrophoresis, Gel, Pulsed-Field
  • Escherichia coli / classification
  • Escherichia coli / genetics*
  • Escherichia coli / pathogenicity
  • Escherichia coli Infections / epidemiology
  • Escherichia coli Infections / microbiology
  • Escherichia coli Infections / veterinary*
  • Genes, Bacterial
  • Genotype
  • Microbial Sensitivity Tests
  • Multilocus Sequence Typing
  • Phenotype
  • Phylogeny
  • Plasmids / metabolism
  • Poultry Diseases / epidemiology*
  • Poultry Diseases / microbiology
  • Serotyping
  • Spain / epidemiology
  • Virulence / genetics

Grants and funding

This work was partially supported by the grants AGL2011- 28836 and AGL2013-47852-R from the Ministerio de Economía y Competitividad (España) and grants CN2012/303 and EM2014/001 (Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia and the European Regional Development Fund, ERDF). Work from LMG is supported by the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) and the European Social Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.