Conjugation frequency of ESBL- and pAmpC- E. coli in broiler chickens in vivo and in vitro

Background Plasmid-mediated conjugation is a major form of horizontal gene transfer (HGT), facilitating dissemination of antimicrobial resistance (AMR) and the emergence of multi-drug-resistant (MDR) strains. In poultry, Escherichia coli producing extended-spectrum β-lactamases (ESBL) and plasmid-mediated AmpC β-lactamase (pAmpC) enzymes are common and contribute to antibiotic resistance. Additionally, plasmid-mediated colistin resistance gene mcr-1 in poultry requires attention, as it is a last-resort antibiotic in human medicine. Although plasmid-mediated conjugation is known to play a role in spreading antimicrobial resistance, its specific impact on resistance transmission within the broiler microbiota is still not well understood. We assessed conjugation dynamics of the mcr-1 gene from a pAmpC- E. coli to an ESBL- E. coli observed in an in vivo broiler chicken trial and compared them to conjugation frequencies under different in vitro conditions (LB broth, intestinal chicken cells in DMEM/F-12 medium, and DMEM/F-12 medium alone), with two initial bacterial loads. Results From in vivo trial, among 138 broiler chickens sampled after a 49-day fattening period, transconjugants were detected in the cecal content of 35 broilers. The median conjugation frequency observed was of -5.02 log10 transconjugants/donor. Median conjugation frequencies across all in vitro conditions varied by less than one log unit (between − 6.8 and − 6.0 log10 transconjugants/donor), and no significant differences in conjugation efficiency were observed between initial bacterial concentrations. Conclusions We confirmed bacterial conjugation between pAmpC-producing E. coli carrying the mcr-1 gene and ESBL-producing E. coli both in vitro and in vivo. The similar conjugation efficiencies observed across different in vitro methods suggest that experimental conditions have minimal influence under controlled settings. In contrast, the in vivo results underscore the significance of the host’s physiological environment in HGT. The presence of transconjugants after a 49-day fattening period indicates that intestinal bacteria function as reservoirs for resistance plasmids and could facilitate their spread throughout the broiler production chain. However, limitations like the possibility of plasmid transfer to other bacteria, unknown persistence of the plasmid in the gut, and potential modulations of transfer efficiency under antibiotic selection must be considered when interpreting the results.