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JB Accepts, published online ahead of print on 17 November 2006
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J. Bacteriol. doi:10.1128/JB.00953-06
Copyright (c) 2006, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Role of a Putative Polysaccharide Locus in Bordetella Biofilm Development

Gina Parise, Meenu Mishra, Yoshikane Itoh, Tony Romeo, and Rajendar Deora*

Program in Molecular Genetics, Department of Microbiology and Immunology, Wake Forest University Health Sciences, Medical Center Blvd., Winston-Salem, NC. 27157.; Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia

* To whom correspondence should be addressed. Email: rdeora{at}wfubmc.edu.


  Abstract

Bordetellae are Gram-negative bacteria which colonize the respiratory tract of animals and humans. We and others have recently shown that these bacteria are capable of living as sessile communities known as biofilms on a number of abiotic surfaces. During the biofilm mode of existence, bacteria produce one or more extracellular polymeric substances which function in part, to hold the cells together and to a surface. There is little information on either the constituents of the biofilm matrix or the genetic basis of biofilm development by Bordetella. By utilizing immunoblot assays and by enzymatic hydrolysis using Dispersin B (DspB), a glycosyl hydrolase that specifically cleaves the polysaccharide, poly-{beta}-1,6-N-acetyl-D-glucosamine (GlcNAc), we provide evidence for the production of poly-{beta}-1,6-GlcNAc by various Bordetella species (B. bronchiseptica, B. pertussis and B. parapertussis) and its role in their biofilm development. We have investigated the role of a Bordetella locus designated herein as bpsABCD (Bordetella polysaccharide) in biofilm formation. The bps locus is homologous to several bacterial loci that are required for the production of poly-{beta}-1,6-GlcNAc and have been implicated in bacterial biofilm formation. By utilizing multiple microscopic techniques to analyze biofilm formation under both static and hydrodynamic conditions, we demonstrate that the bps locus although not essential at initial stages of biofilm formation, contributes to the stability and the maintenance of the complex architecture of Bordetella biofilms.


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