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Journal of Bacteriology, August 2001, p. 4451-4458, Vol. 183, No. 15
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.15.4451-4458.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

The Type IV Fimbrial Subunit Gene (fimA) of Dichelobacter nodosus Is Essential for Virulence, Protease Secretion, and Natural Competence

Ruth M. Kennan,1,* Om P. Dhungyel,2 Richard J. Whittington,3 John R. Egerton,2 and Julian I. Rood1

Bacterial Pathogenesis Research Group, Department of Microbiology, Monash University, Victoria 3800,1 Department of Veterinary Clinical Sciences, The University of Sydney, Camden, New South Wales 2570,2 and Microbiology and Immunology Section, Elizabeth Macarthur Agricultural Institute, NSW Agriculture, Menangle, New South Wales 2568,3 Australia

Received 20 March 2001/Accepted 15 May 2001

Dichelobacter nodosus is the essential causative agent of footrot in sheep. The major D. nodosus-encoded virulence factors that have been implicated in the disease are type IV fimbriae and extracellular proteases. To examine the role of the fimbriae in virulence, allelic exchange was used to insertionally inactivate the fimA gene, which encodes the fimbrial subunit protein, from the virulent type G D. nodosus strain VCS1703A. Detailed analysis of two independently derived fimA mutants revealed that they no longer produced the fimbrial subunit protein or intact fimbriae and did not exhibit twitching motility. In addition, these mutants were no longer capable of undergoing natural transformation and did not secrete wild-type levels of extracellular proteases. These effects were not due to polar effects on the downstream fimB gene because insertionally inactivated fimB mutants were not defective in any of these phenotypic tests. Virulence testing of the mutants in a sheep pen trial conducted under controlled environmental conditions showed that the fimA mutants were avirulent, providing evidence that the fimA gene is an essential D. nodosus virulence gene. These studies represent the first time that molecular genetics has been used to determine the role of virulence genes in this slow growing anaerobic bacterium.

* Corresponding author. Mailing address: Bacterial Pathogenesis Research Group, Department of Microbiology, Monash University, Victoria 3800, Australia. Phone: 613 9905 4808. Fax: 613 9905 4811. E-mail:ruth.kennan{at}med.monash.edu.au.

Journal of Bacteriology, August 2001, p. 4451-4458, Vol. 183, No. 15
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.15.4451-4458.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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