Skip to main content

Conditions for Optimal Candida Biofilm Development in Microtiter Plates

  • Protocol
Candida albicans

Part of the book series: Methods in Molecular Biology ((MIMB,volume 499))

Abstract

Development of Candida spp. biofilms on medical devices such as catheters and voice prosthesis has been recognized as an increasing clinical problem. Simple device removal is often impossible, while in addition, resulting candidal infections are difficult to resolve due to their increased resistance to many antifungal agents. Susceptibility studies of clinical isolates are generally performed according to the CLSI standard, which measures planktonic cell susceptibility, but similar standards have not been designed or applied to testing of cells growing within a biofilm. As consistent biofilms from many strains are more difficult to simultaneously obtain and analyze than are independent planktonic cultures, any standard assay must address these concerns. In the present chapter, optimized conditions that promote biofilm formation within individual wells of microtiter plates are described. In addition, the method has proven useful in preparing C. albicans biofilms for investigation by a variety of microscopic and molecular techniques.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Douglas, L. J. (2002) Medical importance of biofilms in Candida infections. Rev. Iberoam. Micol. 19, 139–143.

    PubMed  Google Scholar 

  2. Kojic, E. M., and Darouiche R. O. (2004) Candida infections of medical devices. Clin. Microbiol. Rev. 17, 255–267.

    Article  PubMed  Google Scholar 

  3. Krom, B. P., Cohen, J. B., McElhaney Feser, G. E., and Cihlar, R. L. (2007) Optimized candidal biofilm microtiter assay. J. Microbiol. Methods. 68(2), 421–423.

    Article  PubMed  CAS  Google Scholar 

  4. Fonzi, W. A., and Irwin, M. Y. (1993) Isogenic strain construction and gene mapping in Candida albicans. Genetics 134, 717–728.

    PubMed  CAS  Google Scholar 

  5. Staab, J. F., Bahn, Y. S., and Sundstrom, P. (2003) Integrative, multifunctional plasmids for hypha-specific or constitutive expression of green fluorescent protein in Candida albicans. Microbiology 149, 2977–2986.

    Article  PubMed  CAS  Google Scholar 

  6. NCCLS. (2002) Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts: Approved Standard—Second Edition. NCCLS document M27-A2. NCCLS, Wayne, Pennsylvania, USA.

    Google Scholar 

  7. Chandra, J., Kuhn, D. M., Mukherjee, P. K., Hoyer, L. L., McCormick, T., and Ghannoum, M. A. (2001). Biofilm formation by the fungal pathogen Candida albicans: development, architecture, and drug resistance. J. Bacteriol. 183, 5385–5394.

    Article  PubMed  CAS  Google Scholar 

  8. Al Fattani, M. A., and Douglas, L. J. (2006) Biofilm matrix of Candida albicans and Candida tropicalis: chemical composition and role in drug resistance. J. Med. Microbiol. 55, 999–1008.

    Article  PubMed  CAS  Google Scholar 

  9. Hawser, S. P., Norris, H., Jessup, C. J., and Ghannoum, M. A. (1998). Comparison of a 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-t etrazolium hydroxide (XTT) colorimetric method with the standardized National Committee for Clinical Laboratory Standards method of testing clinical yeast isolates for susceptibility to antifungal agents. J. Clin. Microbiol. 36, 1450–1452.

    PubMed  CAS  Google Scholar 

  10. Kuhn, D. M., Balkis, M., Chandra, J., Mukherjee, P. K., and Ghannoum, M. A. (2003). Uses and limitations of the XTT assay in studies of Candida growth and metabolism. J. Clin. Microbiol. 41, 506–508.

    Article  PubMed  CAS  Google Scholar 

  11. Hawser, S. P., Baillie, G. S., and Douglas, L. J. (1998) Production of extracellular matrix by Candida albicans biofilms. J. Med. Microbiol. 47, 253–256.

    Article  PubMed  CAS  Google Scholar 

  12. Baillie, G. S., and Douglas, L. J. (2000). Matrix polymers of Candida biofilms and their possible role in biofilm resistance to antifungal agents. J. Antimicrob. Chemother. 46, 397–403.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2009 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Krom, B.P., Cohen, J.B., McElhaney-Feser, G., Busscher, H.J., van der Mei, H.C., Cihlar, R.L. (2009). Conditions for Optimal Candida Biofilm Development in Microtiter Plates. In: Cihlar, R.L., Calderone, R.A. (eds) Candida albicans. Methods in Molecular Biology, vol 499. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-151-6_7

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-151-6_7

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-58829-760-0

  • Online ISBN: 978-1-60327-151-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics