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JAC Advance Access published online on March 8, 2006
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkl057
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© The Author 2006. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received November 15, 2005
Revised February 8, 2006
Accepted February 10, 2006

Original article

Identification of blaCMY-7 and associated plasmid-mediated resistance genes in multidrug-resistant Escherichia coli isolated from dogs at a veterinary teaching hospital in Australia

Hanna E. Sidjabat 1, Kirsty M. Townsend 2, Nancy D. Hanson 3, Jan M. Bell 4, H. W. Stokes 5, Kari S. Gobius 6, Susan M. Moss 7, and Darren J. Trott 7 *

1 School of Veterinary Science, The University of Queensland, Brisbane, QLD 4072, Australia; Medical Faculty of the Christian University of Indonesia (FK-UKI), Cawang Atas, Jakarta, Indonesia
2 School of Veterinary Science, The University of Queensland, Brisbane, QLD 4072, Australia; Present address. Division of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, WA 6150, Australia
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
5 Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, NSW 2109, Australia
6 Food Science Australia, PO Box 3312, Tingalpa DC, QLD 4173, Australia
7 School of Veterinary Science, The University of Queensland, Brisbane, QLD 4072, Australia

* To whom correspondence should be addressed.
Darren J. Trott, E-mail: d.trott{at}uq.edu.au


  Abstract

Objectives: To determine clonality and identify plasmid-mediated resistance genes in 11 multidrug-resistant Escherichia coli (MDREC) isolates associated with opportunistic infections in hospitalized dogs in Australia.

Methods: Phenotypic (MIC determinations, modified double-disc diffusion and isoelectric focusing) and genotypic methods (PFGE, plasmid analysis, PCR, sequencing, Southern hybridization, bacterial conjugation and transformation) were used to characterize, investigate the genetic relatedness of, and identify selected plasmid-mediated antimicrobial resistance genes, in the canine MDREC.

Results: Canine MDRECs were divided into two clonal groups (CG 1 and 2) with distinct restriction endonuclease digestion and plasmid profiles. All isolates possessed blaCMY-7 on an ~93 kb plasmid. In CG 1 isolates, blaTEM, catA1 and class 1 integron-associated dfrA17-aadA5 genes were located on an ~170 kb plasmid. In CG 2 isolates, a second ~93 kb plasmid contained blaTEM and unidentified class 1 integron genes, although a single CG 2 strain carried dfrA5. Antimicrobial susceptibility profiling of E. coli K12 transformed with CG 2 large plasmids confirmed that the blaCMY-7-carrying plasmid did not carry any other antimicrobial resistance genes, whereas the blaTEM/class 1 integron-carrying plasmid carried genes conferring resistance to tetracycline and streptomycin also.

Conclusions: This is the first report on the detection of plasmid-mediated blaCMY-7 in animal isolates in Australia. MDREC isolated from extraintestinal infections in dogs may be an important reservoir of plasmid-mediated resistance genes.

Keywords: multidrug-resistant E. coli; blaCMY-7; blaTEM; class 1 integrons.
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