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Effect of oxygen and growth medium on in vitro biofilm formation by Escherichia coli

Published online by Cambridge University Press:  01 January 2006

L. A. Bjergbæk
Section of Environmental Engineering, Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University, Sohngaardsholmsvej 57, DK-9000 Aalborg, Denmark
J. A. J. Haagensen
Center for Biomedical Microbiology, BioCentrum-DTU, Technical University of Denmark, Soltofts Plads 221, DK-2800 Kgs. Lyngby, Denmark
A. Reisner
Institut für Molekulare Biowissenschaften, Karl-Franzens-Universität Graz, Universitätsplatz 2, A-8010 Graz, Austria
S. Molin
Center for Biomedical Microbiology, BioCentrum-DTU, Technical University of Denmark, Soltofts Plads 221, DK-2800 Kgs. Lyngby, Denmark
P. Roslev*
Section of Environmental Engineering, Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University, Sohngaardsholmsvej 57, DK-9000 Aalborg, Denmark
*Corresponding author: Dr P. Roslev Section of Environmental Engineering Department of Biotechnology Chemistry and Environmental Engineering Aalborg University Sohngaardsholmsvej 57 DK-9000 Aalborg DenmarkT 45 9635 8505 F 45 9814 2555


The effects of oxygen availability on in vitro biofilm formation by an Escherichia coli K-12 strain and 13 clinical E. coli strains were compared. All E. coli strains were capable of forming monospecies biofilm on polystyrene in aerobic media. The K-12 strain produced biofilm in both aerobic glucose minimal medium (ABTG), and aerobic trypticase soy broth (TSB) whereas the majority of the clinical strains produced significant biofilm only in aerobic TSB (9 of 13). In anaerobic media, E. coli K-12 and 9 of the 13 clinical strains were capable of forming biofilm in vitro. Only three clinical strains formed biofilm in anaerobic TSB whereas six clinical strains produced detectable biofilm in anaerobic ABTG. None of the strains tested were capable of forming biofilm in both anaerobic ABTG and anaerobic TSB. Strains that were good biofilm formers in aerobic ABTG also produced the highest amount of biofilm in anaerobic ABTG (R2 = 0.90). Image analysis revealed notable differences in architecture for biofilms grown in the presence and in the absence of oxygen. In aerobic ABTG, the biofilm was dominated by tall, mushroom-shaped microcolonies with pores and channels whereas biofilm in anaerobic ABTG was thinner and less heterogeneous, resulting in reduced maximum thickness and biovolume. Analysis of phospholipid fatty acid (PLFA) profiles from E. coli K-12 and three clinical strains did not reveal a specific pattern associated with the biofilm phenotypes. Interestingly, the clinical E. coli strains adjusted their PLFA composition much more than did E. coli K-12 in response to changes in growth regimens. Collectively, the results indicate that oxygen availability may affect E. coli biofilm formation in minimal and complex media. The results confirm that E. coli K-12 and some clinical E. coli strains are capable of forming in vitro biofilm under anaerobic conditions. However, the data also suggest that this attribute is highly strain dependent and may vary significantly among clinical isolates.

Copyright © Cambridge University Press 2007

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Arneborg, N., Salskov-Iversen, A. S. & Mathiasen, T. E. (1993) The effect of growth rate and other growth conditions on the lipid composition of Escherichia coli. Applied Microbiology and Biotechnology 39, 353357CrossRefGoogle Scholar
Blattner, F. R., Plunkett, G., III, Bloch, C. A., Perna, N. T., Burland, V., Riley, M., et al. (1997) The complete genome sequence of Escherichia coli K-12. Science 277, 14531474CrossRefGoogle ScholarPubMed
Castonguay, M. H., van der Schaaf, S., Koester, W., Krooneman, J., van der Meer, W., Harmsen, H. & Landini, P. (2006) Biofilm formation by Escherichia coli is stimulated by synergistic interactions and co-adhesion mechanisms with adherence- proficient bacteria. Research in Microbiology 157, 471478CrossRefGoogle ScholarPubMed
Christensen, B. B., Sternberg, C., Andersen, J. B., Palmer, R. J. J., Nielsen, A. T., Givskov, M. & Molin, S. (1999) Molecular tools for study of biofilm physiology. Methods in Enzymolology 310, 2042CrossRefGoogle ScholarPubMed
Clark, D. J. & Maaløe, Z. (1967) DNA replication and the division cycle in Escherichia coli. Journal of Molecular Biology 23, 99112CrossRefGoogle Scholar
Colon-Gonzalez, M., Mendez-Ortiz, M. M. & Membrillo-Hernandez, J. (2004) Anaerobic growth does not support biofilm formation in Escherichia coli K-12. Research in Microbiology 155, 514521CrossRefGoogle Scholar
Corona-Izquierdo, F. P. & Membrillo-Hernandez, J. (2002) Biofilm formation in Escherichia coli is affected by 3-(N-morpholino)propane sulfonate (MOPS). Research in Microbiology 153, 181185CrossRefGoogle Scholar
Dewanti, R. & Wong, A. C. L. (1995) Influence of culture conditions on biofilm formation by Escherichia coli O157-H7. International Journal of Food Microbiology 26, 147164CrossRefGoogle ScholarPubMed
Hansen, M. C., Palmer, R. J., Udsen, C., White, D. C. & Molin, S. (2001) Assessment of GFP fluorescence in cells of Streptococcus gordonii under conditions of low pH and low oxygen concentration. Microbiology-Sgm 147, 13831391CrossRefGoogle Scholar
James, B. W. & Keevil, C. W. (1999) Influence of oxygen availability on physiology, verocytotoxin expression and adherence of Escherichia coli O157. Journal of Applied Microbiology 86, 117124CrossRefGoogle ScholarPubMed
Knivett, V. A. & Cullen, J. (1965) Some factors affecting cyclopropane acid formation in Escherichia coli. Journal of Biochemistry 96, 771776CrossRefGoogle ScholarPubMed
Landini, P. & Zehnder, A. J. B. (2002) The global regulatory hns gene negatively affects adhesion to solid surfaces by anaerobically grown Escherichia coli by modulating expression of flagellar genes and lipopolysaccharide production. Journal of Bacteriology 184, 15221529CrossRefGoogle ScholarPubMed
Pratt, L. A. & Kolter, R. (1998) Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type I pili. Molecular Microbiology 30, 285293CrossRefGoogle ScholarPubMed
Reisner, A., Krogfelt, K. A., Klein, B. M., Zechner, E. L. & Molin, S. (2006) In vitro biofilm formation of commensal and pathogenic E. coli strains: impact of environmental and genetic factors. Journal of Bacteriology 188, 35723581CrossRefGoogle ScholarPubMed
Roslev, P., Iversen, N. & Henriksen, K. (1998) Direct fingerprinting of metabolically active bacteria in environmental samples by substrate specific radiolabelling and lipid analysis. Journal of Microbiological Methods 31, 99111CrossRefGoogle Scholar
Salmon, K., Hung, S. P., Mekjian, K., Baldi, P., Hatfield, G. W. & Gunsalus, R. P. (2003) Global gene expression profiling in Escherichia coli K-12. The effects of oxygen availability and FNR. Journal of Biological Chemistry 278, 2983729855CrossRefGoogle Scholar
Valeur, A., Tunlid, A. & Odham, G. (1988) Differences in lipid-composition between free-living and initially adhered cells of a Gram-negative bacterium. Archives of Microbiology 149, 521526CrossRefGoogle Scholar
Van Houdt, R. & Michiels, C. W. (2005) Role of bacterial cell surface structures in Escherichia coli biofilm formation. Research in Microbiology 156, 626633CrossRefGoogle ScholarPubMed
Wimpenny, J. (2000) Structural determinants in biofilm formation. In Biofilms: Recent Advances in their Study and Control, pp. 3549. Edited by Evans, L. V.. Amsterdam: Harwood Academic PublishersGoogle Scholar
Wolfaardt, G. M., Lawrence, J. R., Robarts, R. D., Caldwell, S. J. & Caldwell, D. E. (1994) Multicellular organization in a degradative biofilm community. Applied and Environmental Microbiology 60, 434446Google Scholar
Yuk, H. G. & Marshall, D. L. (2003) Heat adaptation alters Escherichia coli O157:H7 membrane lipid composition and verotoxin production. Applied and Environmental Microbiology 69, 51155119CrossRefGoogle ScholarPubMed