Microbial Ecology

, Volume 18, Issue 1, pp 1–19

Effect of laminar flow velocity on the kinetics of surface recolonization by Mot+ and MotPseudomonas fluorescens

  • Darren R. Korber
  • John R. Lawrence
  • Ben Sutton
  • Douglas E. Caldwell


Computer-enhanced microscopy (CEM) was used to monitor bacteria colonizing the inner surfaces of a 1×3 mm glass flow cell. Image analysis provided a rapid and reliable means of measuring microcolony count, microcolony area, and cell motility. The kinetics of motile and nonmotilePseudomonas fluorescens surface colonization were compared at flow velocities above (120μm sec−1) and below (8μm sec−1) the strain's maximum motility rate (85μm sec−1). A direct attachment assay confirmed that flagellated cells undergo initial attachment more rapidly than nonflagellated cells at high and low flow. During continuous-flow slide culture, neither the rate of growth nor the timing of recolonization (cell redistribution within surface microenvironments) were influenced by flow rate or motility. However, the amount of reattachment of recolonizing cells was both flow and motility dependent. At 8μm sec−1 flow, motility increased reattachment sixfold, whereas at 120μm sec−1 flow, motility increased reattachment fourfold. The spatial distribution of recolonizing cells was also influenced by motility. Motile cells dispersed over surfaces more uniformly (mean distance to the nearest neighbor was 47.0μm) than nonmotile cells (mean distance was 14.2μm) allowing uniform biofilm development through more effective redistribution of cells over the surface during recolonization. In addition, motile cell backgrowth (where cells colonize against laminar flow) occurred four times more rapidly than nonmotile cell backgrowth at low flow (where rate of motility exceeded flow), and twice as rapidly at high flow (where flow exceeded the rate of motility). The observed backgrowth of Mot+ cells against high flow could only have occurred as the result of motile attachment behavior. These results confirm the importance of motility as a behavioral mechanism in colonization and provides an explanation for enhanced colonization by motile cells in environments lacking concentration gradients necessary for chemotactic behavior.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Attridge SR, Rowley D (1983) The role of the flagellum in the adherence ofVibrio cholerae. J Infect Dis 147:864–872PubMedGoogle Scholar
  2. 2.
    Belas MR, Colwell RR (1982) Adsorption kinetics of laterally and polarly flagellatedVibrio. J Bacteriol 151:1568–1580PubMedGoogle Scholar
  3. 3.
    Block SM, Segall JE, Berg HC (1983) Adaptation kinetics in bacterial chemotaxis. J Bacteriol 154:312–324PubMedGoogle Scholar
  4. 4.
    Caldwell DE, Germida JJ (1985) Evaluation of difference imagery for visualizing and quantitating microbial growth. Can J Microbiol 31:35–44Google Scholar
  5. 5.
    Caldwell DE, Lawrence JR (1986) Growth kinetics ofPseudomonas fluorescens microcolonies within the hydrodynamic boundary layers of surface microenvironments. Microb Ecol 12:299–312Google Scholar
  6. 6.
    Davidson AM, Fry JC (1987) A mathematical model for the growth of bacterial microcolonies on marine sediment. Microb Ecol 13:31–45Google Scholar
  7. 7.
    De Weger LD, Van Der Vlugt CIM, Wijfjes AHM, Bakker PAHM, Schippers B, Lugtenberg B (1987) Flagella of a plant-growth-stimulatingPseudomonas fluorescens strain are required for colonization of potato roots. J Bacteriol 169:2769–2773PubMedGoogle Scholar
  8. 8.
    Guentzel MN, Berry LJ (1983) Motility as a virulence factor forVibrio cholerae. Infect Immun 11:890–897Google Scholar
  9. 9.
    Haefele DM, Lindow SE (1987) Flagellar motility confers epiphytic fitness advantages uponPseudomonas syringae. Appl Environ Microbiol 53:2528–2533Google Scholar
  10. 10.
    Ho CS (1986) An understanding of the forces in the adhesion of micro-organisms to surfaces. Process Biochemistry 21:148–152Google Scholar
  11. 11.
    Kefford B, Marshall KC (1986) The role of bacterial surface and substratum hydrophobicity in adhesion ofLeptospira biflexa serovar patoc 1 to inert surfaces. Microb Ecol 12:315–322Google Scholar
  12. 12.
    Kieft TL, Caldwell DE (1983) A computer simulation of surface microcolony formation during microbial colonization. Microb Ecol 9:7–13Google Scholar
  13. 13.
    Kirsop BE, Snell JJS (1984) Maintenance of microorganisms: a manual of laboratory methods. Academic Press, London, pp 35–40Google Scholar
  14. 14.
    Lawrence JR, Caldwell DE (1987) Behavior of bacterial stream populations within the hydrodynamic boundary layers of surface microenvironments. Microb Ecol 14:15–27Google Scholar
  15. 15.
    Lawrence JR, Delaquis PJ, Korber DR, Caldwell DE (1987) Behavior ofPseudomonas fluorescens within the hydrodynamic boundary layers of surface microenvironments. Microb Ecol 14:1–14Google Scholar
  16. 16.
    Marshall KC, Stout R, Mitchell R (1971) Mechanisms of the initial events in the sorption of marine bacteria to solid surfaces. J Gen Microbiol 68:337–348Google Scholar
  17. 17.
    Mayfield CI, Inniss WE (1977) A rapid, simple method for staining bacterial flagella. Can J Microbiol 23:1311–1313PubMedGoogle Scholar
  18. 18.
    Meadows PS (1971) The attachment of bacteria to solid surfaces. Arch Microbiol 75:374–381Google Scholar
  19. 19.
    Savage DC, Fletcher M (eds) (1985) Bacterial adhesion: mechanisms and physiological significance. Plenum Press, New YorkGoogle Scholar
  20. 20.
    Scher FM, Kloepper W, Singleton CA (1985) Chemotaxis of fluorescentPseudomonas spp. to soybean seed exudatesin vitro and in soil. Can J Microbiol 31:570–574Google Scholar
  21. 21.
    Simon R, Priefer U, Puhler A (1983) A broad host range mobilization system forin vivo genetic engineering: transposon mutagenesis in gram-negative bacteria. Bio/Technology 1:784–791Google Scholar
  22. 22.
    Van Loosdrecht MCM, Norde W, Zehnder AJB (1987) Influence of cell surface characteristics on bacterial adhesion to solid supports. Proc 4th Eur Congress on Biotechnology 4:575–580Google Scholar
  23. 23.
    Vesper SJ, Bauer WD (1986) Role of pili (fimbriae) in attachment ofBradyrhizobium japonicum to soybean roots. Appl Environ Microbiol 52:134–141Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1989

Authors and Affiliations

  • Darren R. Korber
    • 1
  • John R. Lawrence
    • 1
  • Ben Sutton
    • 2
  • Douglas E. Caldwell
    • 1
  1. 1.Department of Applied Microbiology and Food ScienceUniversity of SaskatchewanSaskatoonCanada
  2. 2.Forest Biotechnology CentreB.C. Research CorporationVancouverCanada

Personalised recommendations