Real-time Assessment of Interspecies Candida Biofilm Formation

Pedro M.D.S. Abrantes1, Kauthar Behardien1, Charlene W.J. Africa1, *
1 Maternal Endogenous Infections Studies (MEnIS) Research Laboratories, Department of Medical Biosciences, University of the Western Cape, Bellville, South Africa

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© 2022 Abrantes et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Department of Medical Biosciences, University of the Western Cape, Robert Sobukwe Road | Private Bag X17, Bellville 7535, South Africa; Tel: +27 21 9592341; Fax: +27 21 9593125; E-mail:



Candida infections are responsible for increased morbidity and mortality in immunocompromised patients, with Candida having the ability to form highly structured biofilms that protect them from the external environment and the action of antifungals. Few studies have reported on real-time interactions between Candida species.


This study aimed to investigate the interspecies biofilm changes of oral Candida species using the xCELLigence system and the CV staining technique.


This study demonstrated the mixed biofilm interactions of Candida species using an impedance-based biofilm monitoring system and crystal violet staining.


When using the xCELLigence system, the maximum cell index increased in most mixed biofilms, except for the C. glabrata/C. parapsilosis and C. albicans combinations. Bulk biofilm formation measured by CV staining was the highest in C. albicans and C. tropicalis combinations and the lowest in the C. glabrata/C. parapsilosis combination. Pseudohyphae were observed in C. albicans and C. glabrata in combination with C. tropicalis or C. parapsilosis.


This study is the first to report on the real-time interactions of Candida species using the xCELLigence system and suggests that the presence of specific species influences the biofilm formation of commonly isolated Candida species.

Keywords: Candida, Biofilms, Cell adhesion, CV staining, xCELLigence system, Species interactions.