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Journal of Bacteriology, November 2000, p. 6482-6489, Vol. 182, No. 22
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Development and Dynamics of Pseudomonas sp. Biofilms

Tim Tolker-Nielsen,1 Ulla C. Brinch,2 Paula C. Ragas,1 Jens Bo Andersen,1 Carsten Suhr Jacobsen,2 and Søren Molin1,*

Molecular Microbial Ecology Group, Department of Microbiology, The Technical University of Denmark, DK-2800 Lyngby,1 and Department of Geochemistry, Geological Survey of Denmark and Greenland, DK-2400 Copenhagen,2 Denmark

Received 13 April 2000/Accepted 22 August 2000

Pseudomonas sp. strain B13 and Pseudomonas putida OUS82 were genetically tagged with the green fluorescent protein and the Discosoma sp. red fluorescent protein, and the development and dynamics occurring in flow chamber-grown two-colored monospecies or mixed-species biofilms were investigated by the use of confocal scanning laser microscopy. Separate red or green fluorescent microcolonies were formed initially, suggesting that the initial small microcolonies were formed simply by growth of substratum attached cells and not by cell aggregation. Red fluorescent microcolonies containing a few green fluorescent cells and green fluorescent microcolonies containing a few red fluorescent cells were frequently observed in both monospecies and two-species biofilms, suggesting that the bacteria moved between the microcolonies. Rapid movement of P. putida OUS82 bacteria inside microcolonies was observed before a transition from compact microcolonies to loose irregularly shaped protruding structures occurred. Experiments involving a nonflagellated P. putida OUS82 mutant suggested that the movements between and inside microcolonies were flagellum driven. The results are discussed in relation to the prevailing hypothesis that biofilm bacteria are in a physiological state different from planktonic bacteria.

* Corresponding author. Mailing address: Department of Microbiology, Building 301, The Technical University of Denmark, DK-2800 Lyngby, Denmark. Phone: 45 45 25 25 13. Fax: 45 45 88 73 28. E-mail: imsm{at}pop.dtu.dk.

Journal of Bacteriology, November 2000, p. 6482-6489, Vol. 182, No. 22
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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