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Antimicrobial Agents and Chemotherapy, March 2004, p. 838-842, Vol. 48, No. 3
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.3.838-842.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Isolation and Characterization of Integron-Containing Bacteria without Antibiotic Selection

Robert S. Barlow,1,2 John M. Pemberton,2 Patricia M. Desmarchelier,1 and Kari S. Gobius1*

Food Science Australia, Tingalpa DC, Queensland 4173,1 Department of Microbiology and Parasitology, University of Queensland, St. Lucia, Queensland 4072, Australia2

Received 15 April 2003/ Returned for modification 27 July 2003/ Accepted 18 November 2003

The emergence of antibiotic resistance among pathogenic and commensal bacteria has become a serious problem worldwide. The use and overuse of antibiotics in a number of settings are contributing to the development of antibiotic-resistant microorganisms. The class 1 and 2 integrase genes (intI1 and intI2, respectively) were identified in mixed bacterial cultures enriched from bovine feces by growth in buffered peptone water (BPW) followed by integrase-specific PCR. Integrase-positive bacterial colonies from the enrichment cultures were then isolated by using hydrophobic grid membrane filters and integrase-specific gene probes. Bacterial clones isolated by this technique were then confirmed to carry integrons by further testing by PCR and DNA sequencing. Integron-associated antibiotic resistance genes were detected in bacteria such as Escherichia coli, Aeromonas spp., Proteus spp., Morganella morganii, Shewanella spp., and urea-positive Providencia stuartii isolates from bovine fecal samples without the use of selective enrichment media containing antibiotics. Streptomycin and trimethoprim resistance were commonly associated with integrons. The advantages conferred by this methodology are that a wide variety of integron-containing bacteria may be simultaneously cultured in BPW enrichments and culture biases due to antibiotic selection can be avoided. Rapid and efficient identification, isolation, and characterization of antibiotic resistance-associated integrons are possible by this protocol. These methods will facilitate greater understanding of the factors that contribute to the presence and transfer of integron-associated antibiotic resistance genes in bacterial isolates from red meat production animals.

* Corresponding author. Mailing address: Food Science Australia, Cnr Wynnum and Creek Roads, Cannon Hill 4170, Australia. Phone: 61-7-3214-2036. Fax: 61-7-3214-2062. E-mail: Kari.Gobius{at}csiro.au.

Antimicrobial Agents and Chemotherapy, March 2004, p. 838-842, Vol. 48, No. 3
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.3.838-842.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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