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Antimicrobial Agents and Chemotherapy, August 2004, p. 2999-3005, Vol. 48, No. 8
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.8.2999-3005.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Comparison of the Susceptibilities of Burkholderia pseudomallei to Meropenem and Ceftazidime by Conventional and Intracellular Methods

T. J. J. Inglis,1* F. Rodrigues,1 P. Rigby,2 R. Norton,3 and B. J. Currie4

The Division of Microbiology and Infectious Diseases, Western Australian Centre for Pathology and Medical Research, Nedlands,1 Lotteries Confocal Microscopy Centre, Department of Pharmacology, Faculty of Medicine and Dentistry, University of Western Australia, Western Australia,2 QHPS, Townsville General Hospital, Townsville, Queensland,3 The Menzies School of Health Research and Northern Territory Clinical School, Flinders University, Darwin, Northern Territory, Australia4

Received 25 September 2003/ Returned for modification 22 December 2003/ Accepted 23 March 2004

The effect of the two antibiotics ceftazidime and meropenem on a collection of 46 Burkholderia pseudomallei isolates representing clinical and environmental sources across northern Australia was investigated by using a series of in vitro test methods. The susceptibility testing methods used included Kirby-Bauer disk diffusion, Etest MIC, broth microdilution MIC, and a modification of the microdilution method in which Acanthamoeba cells were added to simulate the effect of a professional phagocytic cell on test outcome. In a semiquantitative validation coculture series, the majority of bacteria were intracellular up to a multiplicity of infection of 10 bacteria to one ameba. The optical density and bacterial count (log10 CFU/ml) correlated across the range tested (r2 = 0.77; P < 0.0001). Susceptibility test results were compared against clinical outcomes. The MICs of ceftazidime were consistently higher than those of meropenem by all three methods. The MICs of both agents were significantly higher when Acanthamoeba trophozoites were added to the broth microdilution method. Conventional and intracellular MIC results were consistent for clinical isolates from the Western Australian outbreak cluster despite the wide variety of clinical outcomes. Further development of the intracellular MIC method is expected to help assess the efficacy of antimicrobial agents on this bacterial species in an intracellular setting.

* Corresponding author. Mailing address: The Division of Microbiology and Infectious Diseases, Western Australian Centre for Pathology and Medical Research, Locked Bag 2009, Nedlands, WA 6009, Australia. Phone: 618 9346 3461. Fax: 618 3481 7169. E-mail: tim.inglis{at}health.wa.gov.au.

Antimicrobial Agents and Chemotherapy, August 2004, p. 2999-3005, Vol. 48, No. 8
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.8.2999-3005.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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