Selection and transfer of an IncI1‐tet(A) plasmid of Escherichia coli in an ex vivo model of the porcine caecum at doxycycline concentrations caused by crosscontaminated feed

Aims: The aim of this study was to investigate the effect of subtherapeutic intestinal doxycycline (DOX) concentrations (4 and 1mgl(-1)), caused by cross-contamination of feed, on the enrichment of a DOX-resistant commensal Escherichia coli and its resistance plasmid in an exvivo model of the porcine caecum. Methods and Results: A DOX-resistant, tet(A)-carrying, porcine commensal E.coli strain (EC 682) was cultivated for 6days in the porcine caecum model under different conditions (0, 1 and 4mgl(-1) DOX). EC 682, other coliforms and anaerobic bacteria were enumerated daily. A selection of isolated DOX-resistant coliforms (n=454) was characterized by rep-PCR clustering, PCR assays (Inc1 and tet(A)) and micro broth dilution susceptibility tests (Sensititre). Both 1 and 4mgl(-1) DOX-enriched medium had a significantly higher selective effect on EC 682 and other resistant coliforms than medium without DOX. Transconjugants of EC 682 were isolated more frequently in the presence of 1 and 4mgl(-1) DOX compared to medium without DOX. Conclusions: Subtherapeutic intestinal DOX concentrations have the potential to select for DOX-resistant E.coli, and promote the selection of transconjugants in a porcine caecum model. Significance and Impact of the Study: Cross-contamination of feed with antimicrobials such as DOX likely promotes the spread of antimicrobial resistance. Therefore, it is important to develop or fine-tune guidelines for the safe use of antimicrobials in animal feed and its storage.