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Open Access Highly Accessed Research article

Pseudomonas aeruginosa Cystic Fibrosis isolates of similar RAPD genotype exhibit diversity in biofilm forming ability in vitro

Elena Deligianni1, Sally Pattison2, Daniel Berrar3, Nigel G Ternan1*, Richard W Haylock1, John E Moore4, Stuart J Elborn2 and James SG Dooley1

Author Affiliations

1 Infection and Immunity Research Group, School of Biomedical Sciences, University of Ulster, Cromore Road, Coleraine, BT52 1SA, Northern Ireland, UK

2 School of Medicine, Dentistry and Biomedical Sciences, Queen's University of Belfast, 73 University Road, Belfast, BT7 1NN, Northern Ireland, UK

3 Systems Biology Research Group, School of Biomedical Sciences, University of Ulster, Road, Coleraine, BT52 1SA, Northern Ireland, UK

4 Northern Ireland Public Health Laboratory Service, Department of Bacteriology, Belfast City Hospital, Belfast, BT9 7AD, Northern Ireland, UK

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BMC Microbiology 2010, 10:38  doi:10.1186/1471-2180-10-38

Published: 8 February 2010

Abstract

Background

Pseudomonas aeruginosa is considered to grow in a biofilm in cystic fibrosis (CF) chronic lung infections. Bacterial cell motility is one of the main factors that have been connected with P. aeruginosa adherence to both biotic and abiotic surfaces. In this investigation, we employed molecular and microscopic methods to determine the presence or absence of motility in P. aeruginosa CF isolates, and statistically correlated this with their biofilm forming ability in vitro.

Results

Our investigations revealed a wide diversity in the production, architecture and control of biofilm formation. Of 96 isolates, 49% possessed swimming motility, 27% twitching and 52% swarming motility, while 47% were non-motile. Microtitre plate assays for biofilm formation showed a range of biofilm formation ability from biofilm deficient phenotypes to those that formed very thick biofilms. A comparison of the motility and adherence properties of individual strains demonstrated that the presence of swimming and twitching motility positively affected biofilm biomass. Crucially, however, motility was not an absolute requirement for biofilm formation, as 30 non-motile isolates actually formed thick biofilms, and three motile isolates that had both flagella and type IV pili attached only weakly. In addition, CLSM analysis showed that biofilm-forming strains of P. aeruginosa were in fact capable of entrapping non-biofilm forming strains, such that these 'non-biofilm forming' cells could be observed as part of the mature biofilm architecture.

Conclusions

Clinical isolates that do not produce biofilms in the laboratory must have the ability to survive in the patient lung. We propose that a synergy exists between isolates in vivo, which allows "non biofilm-forming" isolates to be incorporated into the biofilm. Therefore, there is the potential for strains that are apparently non-biofilm forming in vitro to participate in biofilm-mediated pathogenesis in the CF lung.