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Mechanisms of Resistance

Efflux-Mediated Aminoglycoside and Macrolide Resistance in Burkholderia pseudomallei

Richard A. Moore, David DeShazer, Shauna Reckseidler, Ania Weissman, Donald E. Woods
Richard A. Moore
Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, Calgary, Alberta, Canada T2N 4N1
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David DeShazer
Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, Calgary, Alberta, Canada T2N 4N1
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Shauna Reckseidler
Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, Calgary, Alberta, Canada T2N 4N1
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Ania Weissman
Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, Calgary, Alberta, Canada T2N 4N1
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Donald E. Woods
Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, Calgary, Alberta, Canada T2N 4N1
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DOI: 10.1128/AAC.43.3.465
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ABSTRACT

Burkholderia pseudomallei, the causative agent of melioidosis, is intrinsically resistant to a wide range of antimicrobial agents including β-lactams, aminoglycosides, macrolides, and polymyxins. We used Tn5-OT182 to mutagenizeB. pseudomallei to identify the genes involved in aminoglycoside resistance. We report here on the identification of AmrAB-OprA, a multidrug efflux system inB. pseudomallei which is specific for both aminoglycoside and macrolide antibiotics. We isolated two transposon mutants, RM101 and RM102, which had 8- to 128-fold increases in their susceptibilities to the aminoglycosides streptomycin, gentamicin, neomycin, tobramycin, kanamycin, and spectinomycin. In addition, both mutants, in contrast to the parent, were susceptible to the macrolides erythromycin and clarithromycin but not to the lincosamide clindamycin. Sequencing of the DNA flanking the transposon insertions revealed a putative operon consisting of a resistance, nodulation, division-type transporter, a membrane fusion protein, an outer membrane protein, and a divergently transcribed regulator protein. Consistent with the presence of an efflux system, both mutants accumulated [3H]dihydrostreptomycin, whereas the parent strain did not. We constructed an amr deletion strain,B. pseudomallei DD503, which was hypersusceptible to aminoglycosides and macrolides and which was used successfully in allelic exchange experiments. These results suggest that an efflux system is a major contributor to the inherent high-level aminoglycoside and macrolide resistance found inB. pseudomallei.

  • Copyright © 1999 American Society for Microbiology
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Efflux-Mediated Aminoglycoside and Macrolide Resistance in Burkholderia pseudomallei
Richard A. Moore, David DeShazer, Shauna Reckseidler, Ania Weissman, Donald E. Woods
Antimicrobial Agents and Chemotherapy Mar 1999, 43 (3) 465-470; DOI: 10.1128/AAC.43.3.465

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Efflux-Mediated Aminoglycoside and Macrolide Resistance in Burkholderia pseudomallei
Richard A. Moore, David DeShazer, Shauna Reckseidler, Ania Weissman, Donald E. Woods
Antimicrobial Agents and Chemotherapy Mar 1999, 43 (3) 465-470; DOI: 10.1128/AAC.43.3.465
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