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Journal of Clinical Microbiology, October 2005, p. 5247-5255, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.5247-5255.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Heterogeneity of Biofilms Formed by Nonmucoid Pseudomonas aeruginosa Isolates from Patients with Cystic Fibrosis

Department of Clinical Microbiology, Rigshospitalet, and Department of Bacteriology, Institute for Medical Microbiology and Immunology, Panum Institute, University of Copenhagen, DK-2100 Copenhagen, Denmark,¹ Center for Biomedical Microbiology, BioCentrum-DTU, Building 301, Technical University of Denmark, DK-2800 Lyngby, Denmark,² Department of Microbiological Food Safety, Danish Institute for Food and Veterinary Research, Moerkhoej Bygade 19, 2860 Soeborg, Denmark³

Received 27 February 2005/ Returned for modification 17 April 2005/ Accepted 5 July 2005

Biofilms are thought to play a key role in the occurrence of lung infections by Pseudomonas aeruginosa in patients with cystic fibrosis (CF). In this study, 20 nonmucoid P. aeruginosa isolates collected during different periods of chronic infection from eight CF patients were assessed with respect to phenotypic changes and in vitro biofilm formation. The physiological alterations were associated with a loss of motility (35% were nonmotile) and with decreased production of virulence factors (pyocyanin, proteases) and quorum-sensing molecules (45% of the isolates were unable to produce 3-O-C₁₂-homoserine lactone quorum-sensing molecules). Compared with wild-type strain PAO1, most P. aeruginosa isolates demonstrated different degrees of reduction of adherence on polystyrene surfaces. The in vitro biofilm formation of isolates was investigated in a hydrodynamic flow system. Confocal laser scanning microscope analysis showed that the biofilm structures of the P. aeruginosa isolates were highly variable in biomass and morphology. Biofilm development of six genotypically identical sequential isolates recovered from a particular patient at different time points of chronic infection (20 years) and after lung transplantation demonstrated significant changes in biofilm architectures. P. aeruginosa biofilm formation followed a trend of decreased adherence with progression of the chronic lung infection. The results suggest that the adherent characteristic of in vitro biofilm development was not essential for the longitudinal survival of nonmucoid P. aeruginosa during chronic lung colonization.

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