Escherichia coli belonging to ST131 rarely transfers blactx-m-15 to fecal Escherichia coli

Abstract

Karen Rønø Thingholm,1 Frederik Boëtius Hertz,1,2 Anders Løbner-Olesen,3 Niels Frimodt-Møller,1 Karen Leth Nielsen11Department of Clinical Microbiology, Rigshospitalet, Copenhagen 2100, Denmark; 2Department of Clinical Microbiology, Herlev and Gentofte Hospital, Herlev 2730, Denmark; 3Department of Biology, Section for Functional Genomics and Center for Bacterial Stress Response and Persistence, University of Copenhagen, Copenhagen, DenmarkBackground: Extended spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli) causing urinary tract infections often belong to sequence type 131 (ST131), serotype O25, carrying blaCTX-M-15.Aim: The main aim of this study was to examine the conjugational frequencies of E. coli with plasmids carrying blaCTX-M-15 to E. coli isolates from the fecal flora of healthy humans to determine whether ST131 is more likely to uptake or donate ESBL resistance compared to other E. coli clones.Methods:  Donors and recipients were all clinical isolates and did not harbor plasmids with identical incompatibility groups (Inc-groups) based on in silico analyses of Inc-groups and restriction/modification systems (R/M-systems). The in vitro conjugation experiments were performed as filter conjugation with verification of transconjugants by random amplified polymorphic DNA (RAPD) PCR and blaCTX-M-15 PCR.Results: The frequencies of conjugation with blaCTX-M-15-carrying plasmids were found to be very rare with detectable conjugation frequencies in the range of 4x10−9–7x10−7 transconjugants/recipient. Recipients of O25/ST131 type yielded significantly lower conjugation frequencies compared to recipients of other O-types (P=0.004). The applied ST131/O25 donors did not yield detectable levels of transconjugants regardless of the applied recipient. Presence of sub-MIC levels of ampicillin increased plasmid transfer frequencies x100 fold (P=0.07).Conclusion: The results indicate that blaCTX-M-15 is rarely transferred by conjugation to E. coli isolates of the intestinal flora, even when the gene is plasmid-borne.Keywords: horizontal gene transfer, antimicrobial resistance, Escherichia coli, molecular typing, ST131, faecal isolates

Keywords

• Horizontal gene transfer
• antimicrobial resistance
• Escherichia coli
• molecular typing
• ST131
• faecal isolates