Selection of CMY-2 producing Escherichia coli in the faecal flora of dogs treated with cephalexin

Cephalexin is a first generation cephalosporin commonly used in dogs for treatment of pyoderma. The objective of this study was to evaluate the in vivo effects of cephalexin on selection of Escherichia coli resistant to extended-spectrum cephalosporins. A cohort study was conducted on 13 dogs presenting clinical signs of pyoderma and treated with cephalexin and 22 healthy dogs that had not been treated with antibiotics during the previous six months. Selective plating of faeces on MacConkey agar plates containing cefotaxime (CTX) yielded growth of CTX-resistant E. coli for eight of the 13 treated dogs (62%), whereas no growth was observed for any of the control dogs (Fisher exact test, P < 0.001). PCR and sequence analysis identified blaCMY-2 in all eight dogs. PCR-based replicon typing and restriction fragment length polymorphism (RFLP) of E. coli transformants revealed location of blaCMY-2 on indistinguishable IncI1 plasmids in five of the eight dogs. One representative of these five epidemiologically related IncI1 plasmids was further characterized as sequence type (ST2) by plasmid multilocus sequence typing (pMLST). E. coli from the remaining three dogs harboured blaCMY-2 on distinct plasmids with non-typeable replicons. A single isolate was classified as an extraintestinal pathogenic E. coli (ExPEC) due to the presence of iutA, papC and sfa/foc. The results provide a strong indication that cephalexin selects for E. coli producing plasmid-borne CMY-2 β-lactamase. The isolation of a specific IncI1 plasmid carrying blaCMY-2 from five epidemiologically unrelated dogs suggests that cephalexin use may contribute to the spread of this plasmid lineage among Danish dogs.