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Antimicrobial Agents and Chemotherapy, November 2002, p. 3522-3531, Vol. 46, No. 11
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.11.3522-3531.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Action of Chlorhexidine Digluconate against Yeast and Filamentous Forms in an Early-Stage Candida albicans Biofilm

Peter A. Suci1* and Bonnie J. Tyler2

Center for Biofilm Engineering and Microbiology Department, Montana State University, Bozeman, Montana 59717-3980,1 Chemical and Fuels Engineering, University of Utah, Salt Lake City, Utah 841122

Received 17 June 2002/ Returned for modification 28 July 2002/ Accepted 14 August 2002

An in situ method for sensitive detection of differences in the action of chlorhexidine against subpopulations of cells in Candida albicans biofilms is described. Detection relies on monitoring the kinetics of propidium iodide (PI) penetration into the cytoplasm of individual cells during dosing with chlorhexidine. Accurate estimation of the time for delivery of the dosing concentration to the substratum was facilitated by using a flow cell system for which transport to the interfacial region was previously characterized. A model was developed to quantify rates of PI penetration based on the shape of the kinetic data curves. Yeast were seeded onto the substratum, and biofilm formation was monitored microscopically for 3 h. During this period a portion of the yeast germinated, producing filamentous forms (both hyphae and pseudohyphae). When the population was subdivided on the basis of cell morphology, rates of PI penetration into filamentous forms appeared to be substantially higher than for yeast forms. Based on the model, rates of penetration were assigned to individual cells. These data indicated that the difference in rates between the two subpopulations was statistically significant (unpaired t test, P < 0.0001). A histogram of rates and analysis of variance indicated that rates were approximately equally distributed among different filamentous forms and between apical and subapical segments of filamentous forms.

* Corresponding author. Mailing address: Center for Biofilm Engineering and Microbiology Department, Montana State University, Bozeman, MT 59717-3980. Phone: (406) 994-4564. Fax: (406) 994-6098. E-mail: peter_s{at}erc.Montana.edu.

Antimicrobial Agents and Chemotherapy, November 2002, p. 3522-3531, Vol. 46, No. 11
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.11.3522-3531.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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