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Infection and Immunity, January 2004, p. 133-144, Vol. 72, No. 1
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.1.133-144.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Cross-Sectional Analysis of Clinical and Environmental Isolates of Pseudomonas aeruginosa: Biofilm Formation, Virulence, and Genome Diversity

Nathan E. Head and Hongwei Yu*

Department of Microbiology, Immunology and Molecular Genetics, Joan C. Edwards School of Medicine, Marshall University, Huntington, West Virginia 25704-9330

Received 4 April 2003/ Returned for modification 14 July 2003/ Accepted 23 September 2003

Chronic lung infections with Pseudomonas aeruginosa biofilms are associated with refractory and fatal pneumonia in cystic fibrosis (CF). In this study, a group of genomically diverse P. aeruginosa isolates were compared with the reference strain PAO1 to assess the roles of motility, twitching, growth rate, and overproduction of a capsular polysaccharide (alginate) in biofilm formation. In an in vitro biofilm assay system, P. aeruginosa displayed strain-specific biofilm formation that was not solely dependent on these parameters. Compared with non-CF isolates, CF isolates expressed two opposing growth modes: reduced planktonic growth versus efficient biofilm formation. Planktonic cells of CF isolates showed elevated sensitivity to hydrogen peroxide, a reactive oxygen intermediate, and decreased lung colonization in an aerosol infection mouse model. Despite having identical genomic profiles, CF sequential isolates produced different amounts of biofilm. While P. aeruginosa isolates exhibited genomic diversity, the genome size of these isolates was estimated to be 0.4 to 19% (27 to 1,184 kb) larger than that of PAO1. To identify these extra genetic materials, random amplification of polymorphic DNA was coupled with PAO1-subtractive hybridization. Three loci were found within the genomes of two CF isolates encoding one novel homolog involved in retaining a Shigella virulence plasmid (mvpTA) and two divergent genes that function in removing negative supercoiling (topA) and biosynthesis of pyoverdine (PA2402). Together, P. aeruginosa biodiversity could provide one cause for the variation of morbidity and mortality in CF. P. aeruginosa may possess undefined biofilm adhesins that are important to the development of an antibiofilm therapeutic target.

* Corresponding author: Mailing address: Department of Microbiology, Immunology and Molecular Genetics, Marshall University School of Medicine, 1542 Spring Valley Dr., Huntington, WV 25704-9330. Phone: (304) 696-7356. Fax: (304) 696-7207. E-mail: yuh{at}marshall.edu.

Editor: V. J. DiRita

Infection and Immunity, January 2004, p. 133-144, Vol. 72, No. 1
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.1.133-144.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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