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Infection and Immunity, April 2003, p. 1622-1629, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.1622-1629.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Flagella Are Virulence Determinants of Burkholderia pseudomallei

K. L. Chua,* Y. Y. Chan, and Y. H. Gan

Department of Biochemistry, National University of Singapore, Faculty of Medicine, Singapore 119260

Received 11 June 2002/ Returned for modification 8 October 2002/ Accepted 23 December 2002

Burkholderia pseudomallei, a facultatively intracellular pathogen, is a flagellated and motile gram-negative bacterium and is the causative agent of melioidosis in humans. Flagella are commonly recognized as important virulence determinants expressed by bacterial pathogens since the motility phenotype imparted by these organelles often correlates with the ability of an organism to cause disease. We used a virulent isolate of B. pseudomallei, KHW, to construct an isogenic deletion mutant with a mutation in the flagellin gene (fliC) by gene replacement transposon mutagenesis. The KHW{Delta}fliCKm mutant was aflagellate and nonmotile in semisolid agar. The isogenic KHW{Delta}fliCKm mutant was not impaired in terms of the ability to invade and replicate in cultured human lung cells compared with the wild type. It was also equally virulent in slow-killing assays involving Caenorhabditis elegans, but it was avirulent during intranasal infection of BALB/c mice. Very few bacteria, if any, were isolated from the lungs and spleens of KHW{Delta}fliCKm-infected mice. In contrast, the bacterial loads in the lungs and spleens were similar in mice infected with KHW and in mice infected with the complemented mutant, KHW{Delta}fliCKm/pUCP28TfliC. Unlike the Syrian hamster or diabetic rat models of infection, the B. pseudomallei flagellin was also a virulence factor during intraperitoneal infection of BALB/c mice. In this study, all animals infected with KHW{Delta}fliCKm remained healthy and did not succumb to disease regardless of the route of infection. The flagellum is therefore an important and necessary virulence determinant of B. pseudomallei during intranasal and intraperitoneal infection of mice.

* Corresponding author. Mailing address: Department of Biochemistry, National University of Singapore, Faculty of Medicine, 10 Kent Ridge Crescent, Singapore 119260. Phone: 65-8743684. Fax: 65-7791453. E-mail: bchckl{at}nus.edu.sg.

Editor: V. J. DiRita

Infection and Immunity, April 2003, p. 1622-1629, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.1622-1629.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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