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J Med Microbiol 56 (2007), 426-434; DOI: 10.1099/jmm.0.46888-0
© 2007 Society for General Microbiology
ISSN 1473-5644

Identification of plasmid-mediated extended-spectrum and AmpC ß-lactamases in Enterobacter spp. isolated from dogs

Hanna E. Sidjabat1,2, Nancy D. Hanson3, Ellen Smith-Moland3, Jan M. Bell4, Justine S. Gibson1, Lucio J. Filippich1 and Darren J. Trott1

1 School of Veterinary Science, The University of Queensland, Brisbane, QLD 4072, Australia

2 Medical Faculty of the Christian University of Indonesia (FK-UKI), Cawang Atas, Jakarta, Indonesia

3 Center for Research in Anti-Infectives and Biotechnology, Department of Medical Microbiology and Immunology, School of Medicine, Creighton University, Omaha, Nebraska, USA

4 Microbiology and Infectious Diseases, Women's and Children's Hospital, North Adelaide, SA 5006, Australia

Correspondence
Darren J. Trott
d.trott{at}uq.edu.au

Received 14 August 2006
Accepted 27 October 2006

The genetic determinants involved in reduced susceptibility to third-generation cephalosporins and aztreonam were identified in ten canine Enterobacter isolates associated with opportunistic infections in three veterinary hospitals in Brisbane, Australia. All isolates were evaluated by a combination of phenotypic (broth microdilution and disc susceptibility, modified disc diffusion and IEF) and genotypic (PFGE, plasmid analysis, Southern blot hybridization, bacterial conjugation, PCR and sequencing) methods to investigate genetic relatedness and to identify plasmid-mediated resistance genes, in particular ß-lactamase genes responsible for extended-spectrum cephalosporin resistance. The ten canine isolates were genotypically diverse based on PFGE and belonged to either Enterobacter cloacae or Enterobacter hormaechei on the basis of 16S rRNA gene sequence analysis. Plasmid profiles were also diverse. Nine isolates contained a transmissible blaSHV-12-carrying plasmid (~140 kb) that also conferred resistance to chloramphenicol, gentamicin, spectinomycin, tetracycline, trimethoprim and sulfonamides. In all plasmid-mediated extended-spectrum ß-lactamase (ESBL)-producing isolates including transconjugants, blaSHV-12 was shown to reside in a ~6.5 kb plasmid fragment. The remaining isolate that was not an ESBL producer possessed an AmpC ß-lactamase gene (blaCMY-2) on a ~93 kb transmissible plasmid. This plasmid did not contain any other antimicrobial resistance genes. Additional plasmid-mediated ß-lactamases identified in some isolates included blaTEM and blaOXA-10. This is the first report of canine Enterobacter isolates containing transmissible plasmid-mediated blaSHV-12 and blaCMY-2 resistance genes. Therefore, Enterobacter isolated from opportunistic infections in dogs may be an important reservoir of plasmid-mediated resistance genes, which could potentially be spread to other members of the Enterobacteriaceae.

Abbreviations: ESBL, extended-spectrum ß-lactamase; MDR, multidrug-resistant; MDREC, MDR Escherichia coli.






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