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Journal of Clinical Microbiology, September 2001, p. 3234-3240, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3234-3240.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Biofilm Formation by Candida dubliniensis

Gordon Ramage,1 Kacy Vande Walle,2 Brian L. Wickes,1 and José L. López-Ribot2,*

Department of Microbiology,1 and Department of Medicine, Division of Infectious Diseases,2 The University of Texas Health Science Center at San Antonio, San Antonio, Texas

Received 23 March 2001/Accepted 14 June 2001

Candida dubliniensis is an opportunistic yeast closely related to Candida albicans that has been recently implicated in oropharyngeal candidiasis in human immunodeficiency virus-infected patients. Most manifestations of candidiasis are associated with biofilm formation, with cells in biofilms displaying properties dramatically different from free-living cells grown under normal laboratory conditions. Here, we report on the development of in vitro models of C. dubliniensis biofilms on the surfaces of biomaterials (polystyrene and acrylic) and on the characteristics associated with biofilm formation by this newly described species. Time course analysis using a formazan salt reduction assay to monitor metabolic activities of cells within the biofilm, together with microscopy studies, revealed that biofilm formation by C. dubliniensis occurred after initial focal adherence, followed by growth, proliferation, and maturation over 24 to 48 h. Serum and saliva preconditioning films enhanced the initial attachment of C. dubliniensis and subsequent biofilm formation. Scanning electron microscopy and confocal scanning laser microscopy were used to further characterize C. dubliniensis biofilms. Mature C. dubliniensis biofilms consisted of a dense network of yeasts cells and hyphal elements embedded within exopolymeric material. C. dubliniensis biofilms displayed spatial heterogeneity and an architecture showing microcolonies with ramifying water channels. Antifungal susceptibility testing demonstrated the increased resistance of sessile C. dubliniensis cells, including the type strain and eight different clinical isolates, against fluconazole and amphotericin B compared to their planktonic counterparts. C. dubliniensis biofilm formation may allow this species to maintain its ecological niche as a commensal and during infection with important clinical repercussions.

* Corresponding author. Mailing address: Department of Medicine/Div. Infectious Diseases, The University of Texas Health Science Center at San Antonio. South Texas Centers for Biology in Medicine, Texas Research Park, 15355 Lambda Dr., San Antonio, TX 78245. Phone: (210) 562-5017. Fax: (210) 562-5016. E-mail: ribot{at}uthscsa.edu.

Journal of Clinical Microbiology, September 2001, p. 3234-3240, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3234-3240.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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