Examination of the horizontal gene transfer dynamics of an integrative and conjugative element encoding multidrug resistance in Histophilus somni

Abstract

Integrative and conjugative elements (ICEs) are self-transferable mobile genetic elements that play a significant role in disseminating antimicrobial resistance between bacteria via horizontal gene transfer (HGT). A recently identified ICE in a clinical isolate of Histophilus somni (ICEHs02) is 72,914 base-pairs in length and harbours seven predicted antimicrobial resistance genes (ARG) conferring resistance to tetracycline (tetR-tet(H)), florfenicol (floR), sulfonamide (Sul2), aminoglycoside (APH(3'')-Ib), (APH(6)-Id), (APH(3')-Ia), and copper (mco). This study investigated ICEHs02 host range, assessed effects of antimicrobial stressors on transfer frequency, and examined effects of ICEHs02 acquisition on hosts. Conjugation assays examined transfer frequency of ICEHs02 to Histophilus somni and Pasteurella multocida strains. PCR assays confirmed the presence of a circular intermediate, ICE-associated core genes, and cargo genes in recipient strains. Susceptibility testing examined ICEHs02-associated resistance phenotypes in recipient strains. Tetracycline and ciprofloxacin induction significantly increased the transfer rates of ICEHs02 in vitro. The copy numbers of the circular intermediate of ICEHs02 per chromosome exhibited significant increases of ~ 37-fold after tetracycline exposure, and ~ 4-fold after ciprofloxacin treatment. Acquisition of ICEHs02 reduced relative fitness of H. somni transconjugants by 28% (w = 0.72 ± 0.04) and P. multocida TG relative fitness was decreased 15% (w = 0.85 ± 0.01).