Abstract
Although Candida biofilms have been clearly identified as playing an increasingly important role in human disease, their biology and the reason for their poor susceptibility to antifungal agents remain largely unknown. Over recent years, various models have been developed in order to better characterize Candida biofilms. Here, we describe a number of rapid, inexpensive microtiter-format techniques and strategies which can be used for large-scale screening procedures aimed at identifying genes involved in Candida biofilm formation and/or potential antifungal agents with activity against pathogen cells growing under these conditions. The procedures could also be easily adapted for studying biofilm structures with a range of microscopy techniques.
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References
Donlan R.M. and Costerton J.W. (2002) Biofilms: survival mechanisms of clinical relevant microorganisms. Clin Microbiol Rev 15, 167–193.
Costerton J.W., Stewart P.S., and Greenberg E.P. (1999) Bacterial biofilms: a common cause of persistent infections. Science 284, 1318–1322.
Goldmann D.A. and Pier G.B. (1993) Pathogenesis of infections related to intravascular catheterization. Clin Microbiol Rev 6, 176–192.
Douglas L.J. (2003) Candida biofilm and their role infection. Trends Microbiol 11, 30–36.
García-Sanchez S., Aubert S., Iraqui I., Janbon G., Ghigo J.M., and d’Enfert C. (2004) Transcriptome invariance in biofilm populations of Candida albicans. Eukaryot Cell 3, 536–545.
Ramage G., VandeWalle K., Wickes B.L., and Lopez-Ribot J.L. (2001) Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms. Antimicrob Agents Chemother 45, 2475–2479.
Andes D., Nett J., Oschel P., Albrecht R., Marchillo K., and Pitula A. (2004) Development and characterization of an in vivo central venous catheter Candida albicans biofilm model. Infect Immun 72, 6023–6031.
Ricicova M., Kucharikova S., Tournu H., Hendrix J., Bujdakova H., Van Eldere J., Lagrou K., and Van Dijck P. (2010) Candida albicans biofilm formation in a new in vivo rat model. Microbiology 156, 909–919.
Schinabeck M.K., Long L.A., Hossain M.A., Chandra J., Mukherjee P.K., Mohamed S., and Ghannoum M.A. (2004) Rabbit model of Candida albicans biofilm infection: liposomal amphotericin B antifungal lock therapy. Antimicrob Agents Chemother 48, 1727–1732.
Chandra J., Patel J.D., Li J., Zhou G., Mukherjee P.K., McCormick T.S., Anderson J.M., and Ghannoum M.A. (2005) Modification of surface properties of biomaterials influences the ability of Candida albicans to form biofilms. Appl Environ Microbiol 71, 8795–8801.
Nobile C.J. and Mitchell A.P. (2005) Regulation of cell-surface genes and biofilm formation by the C. albicans transcription factor Bcr1p. Curr Biol 15, 1150–1155.
Nobile C.J., Nett J.E., Hernday A.D., Homann O.R., Deneault J.S., Nantel A., Andes D.R., Johnson A.D., and Mitchell A.P. (2009) Biofilm matrix regulation by Candida albicans Zap1. PLoS biology 7, e1000133.
Iraqui I., Garcia-Sanchez S., Aubert S., Dromer F., Ghigo J.-M., d’Enfert C., and Janbon G. (2005) The Yak1p kinase controls expression of adhesins and biofilm formation in Candida glabrata in a Sir4p dependent pathway. Mol Microbiol 55, 1259–1271.
Kumamoto C.A. (2005) In contact-activated kinase signals Candida albicans invasive growth and biofilm development. Proc Natl Acad Sci USA 102, 5576–5581.
Bachmann S.P., VandeWalle K., Ramage G., Patterson T.F., Wickes B.L., Graybill J.R., and Lopez-Ribot J.L. (2002) In vitro activity of caspofungin against Candida albicans biofilms. Antimicrob Agents Chemother 46, 3591–3596.
Kuhn D.M., George T., Chandra J., Mukherjee P.K., and Ghannoum M.A. (2002) Antifungal susceptibility of Candida biofilms: unique efficacy of amphotericin B lipid formulations and echinocandins. Antimicrob Agents Chemother 46, 1773–1780.
Ramage G. and Lopez-Ribot J.L. (2005) Techniques and antifungal susceptibility testing of Candida albicans biofilms. Methods Mol Med 118, 71–79.
Hawser S.P. and Douglas L.J. (1994) Biofilm formation by Candida species on the surface of catheter materials in vitro. Infect Immun 62, 915–921.
Ramage G., Wickes B.L., and Lopez-Ribot J.L. (2001) Biofilms of Candida albicans and their associated resistance to antifungal agent. Am Clin Lab 20, 42–44.
Cagle G.D. (1974) Critical-Point Drying: Rapid method for the determination of bacterial extracellular polymer and surface structures. Appl Microbiol 28, 312–316.
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Riera, M., Moreno-Ruiz, E., Goyard, S., d’Enfert, C., Janbon, G. (2012). Biofilm Formation Studies in Microtiter Plate Format. In: Brand, A., MacCallum, D. (eds) Host-Fungus Interactions. Methods in Molecular Biology, vol 845. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-61779-539-8_25
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DOI: https://doi.org/10.1007/978-1-61779-539-8_25
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