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Antimicrobial Agents and Chemotherapy, May 1999, p. 1034-1041, Vol. 43, No. 5
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Assessment of the Effect of Amphotericin B on the Vitality of Candida albicans

Robert S. Liao, Robert P. Rennie,* and James A. Talbot

Department of Medical Microbiology and Immunology, University of Alberta, Walter MacKenzie Centre, Edmonton, Alberta T6G 2J2, Canada

Received 15 October 1998/Returned for modification 7 January 1999/Accepted 3 March 1999

The processes involved in cell death are complex, and individual techniques measure specific fractions of the total population. The interaction of Candida albicans with amphotericin B was measured with fluorescent probes with different cellular affinities. These were used to provide qualitative and quantitative information of physiological parameters which contribute to fungal cell viability. SYBR Green I and 5,(6)-carboxyfluorescein were used to assess membrane integrity, and bis-(1,3-dibutylbarbituric acid)trimethine oxonol and 3,3-dihexyloxacarbocyanine iodide were used to evaluate alterations in membrane potential. The fluorescent indicators were compared with replication competency, the conventional indicator of viability. By using these tools, the evaluation of the response of C. albicans to amphotericin B time-kill curves delineated four categories which may represent a continuum between alive and dead. The data showed that replication competency (CFU per milliliter) as determined by conventional antifungal susceptibility techniques provided only an estimate of inhibition. Interpretation of fluorescent staining characteristics indicated that C. albicans cells which were replication incompetent after exposure to greater than 0.5 µg of amphotericin B per ml still maintained degrees of physiological function.

* Corresponding author. Mailing address: Department of Microbiology and Public Health, 2B3.08 Walter MacKenzie Centre, University of Alberta Hospital, 8440-112 St., Edmonton, Alberta, Canada T6G 2J2. Phone: (780) 407-4461. Fax: (780) 407-3864. E-mail: rpr{at}bugs.uah.ualberta.ca.

Antimicrobial Agents and Chemotherapy, May 1999, p. 1034-1041, Vol. 43, No. 5
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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