Novel insertion sequence ISChh1-like mediating acquisition of optrA gene in foodborne pathogen Campylobacter coli of swine origin

Abstract

The optrA gene encodes an ABC-F protein which confers cross-resistance to oxazolidinones and phenicols, and its transmission has so far been associated with multiple transposable elements, including IS1216E, prophages, ICEs and Tn558. Here, we identified an optrA gene flanked by two copies of a novel insertion sequence ISChh1-like in the same direction in Campylobacter coli. Seven optrA-positive C. coli were identified from 263 Campylobacter isolates obtained from one swine farm and two slaughterhouses in Sichuan province of China. The optrA genes in 6 isolates were directly flanked by two copies of ISChh1-like elements in the same orientation, in the remaining one isolate, optrA was co-located with fexA and flanked by two copies of IS1216E and inserted in the downstream of the aadE-sat4-aphA3 gene cluster on chromosome. Cloning of optrA into C. jejuni NCTC 11,168 confirmed its role in elevated MICs to oxazolidinones and phenicols. Translocatable units (TUs) and natural transformants were only detected from the isolate with optrA bracketed by IS1216E, not from these with optrA bracketed by ISChh1-like. ISChh1-like in all isolates inserts specifically either next to or between adjacent GG nucleotides, neither have terminal inverted repeats nor generate target site duplications. Interestingly, ISChh1-like were also found mediating integration of other antibiotic resistance genes in Campylobacter, including tet(O), aphA3 and aadE-sat4-aphA3 gene cluster. Taken together, these results identify ISChh1-like as a novel transposon mediating acquisition of multiple antibiotic resistance genes in Campylobacter, including the very important optrA gene, suggesting that it plays an essential role in the transmission of antibiotic resistance genes to Campylobacter.