Prevalence and Epidemiological Relationship of CMY-2 AmpC β-Lactamase and CTX-M Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolates from Broiler Farms in Japan

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To evaluate the prevalence of extended-spectrum cephalosporin (ESC)-resistant Enterobacteriaceae in broiler chickens, 41 rectal samples taken from four commercial farms were examined. Desoxycholate hydrogen sulfide lactose agars,supplemented with either 4 μg/ml cefotaxime or 16 μg/ml ceftazidime, were used to screen ESC-resistant bacteria. ESC-resistant bacteria were isolated from all samples. Of the 164 ESC-resistant bacteria　(included four isolates per a sample), 163 were Escherichia coli, while one isolate was identified as Enterobacter cloacae. Extended-spectrum β-lactamase (ESBL) genes and plasmid-mediated AmpC β-lactamase genes in the isolates were determined by PCR and sequencing. One AmpC β-lactamase gene, blaCMY-2 (66%), and four ESBL genes, blaCTX-M-1 (26%), blaCTX-M-55 (10%), blaSHV-5 (4%) and blaCTX-M-2 (3%), were detected in the E. coli isolates.The epidemiological relationship of the CMY-2 and CTX-M β-lactamase-producing isolates among the farms was analyzed by pulsed-field gel electrophoresis using the XbaI restriction enzyme. Forty-one (Y1–Y41) and 14 (X1–X14) clusters were found in the CMY-2 and CTX-M-carrying E. coli isolates, respectively. Some clusters included isolates derived from more than one farm, indicating some cross-contamination of clonal strains, and spread of CMY-2 AmpC β-lactamase or CTX-M ESBL among the farms.