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Updated: Mar 6, 2026
A Soluble Tetrazolium-Based Reduction Assay to Evaluate the Effect of Antibodies on Candida tropicalis Biofilms
Published on: September 16, 2022
Biofilm formation directly correlates with cell viability in Candida tropicalis on polypropylene
Kavyasree Marabanahalli Yogendraiah1, Bindu Sadanandan2, Lokesh Kyathsandra Natraj1
1Department of Biotechnology, M S Ramaiah Institute of Technology, Bengaluru, 560054, Karnataka, India.
Abstract:
Candida tropicalis, the most prevalent non-Candida albicans Candida species, is an emerging pathogen forming robust biofilms on medical devices, contributing to biofouling, virulence, and antifungal resistance. In this study, growth conditions for six C. tropicalis clinical isolates (C4, U873, U951, U1179, U1309, U1360) and a standard strain (MTCC-184) were optimized on polypropylene using central composite design-based response surface methodology. The parameters tested included temperature, pH, shaker speed, inoculum size, and incubation time, with biofilm formation quantified by crystal violet, cell viability by MTT, biomass by calcofluor white, and wet/dry weight measurements. Notably, C. tropicalis forms biofilm on polypropylene surfaces, resembling extracellular polymeric substance-rich matrices. Among the isolates, C4, U873, U951, and U1179 fit the CCD model, whereas for MTCC-184, U1309, and U1360, the Johnson Transformation was required to obtain unified optimal conditions. Temperature and pH were the major factors influencing biofilm formation in C4 and U1179, while temperature and incubation time were significant for U873 and U951. A direct correlation was observed between cell viability and biofilm formation, though biomass varied, indicating strain-specific virulence. This high-throughput optimization strategy establishes a platform for antifungal screening, biofilm-material interaction studies, and the development of medical devices resistant to fungal colonization. KEY POINTS: • Optimized growth conditions of Candida tropicalis biofilm on polypropylene material by RSM • Four C. tropicalis isolates fit the CCD model; the other three isolates were modelled using CCD-JT • A direct correlation was observed between cell viability and biofilm with variations in cell mass.
Insights
Optimized growth conditions for Candida tropicalis biofilms on polypropylene were determined using response surface methodology. This research aids in developing antifungal strategies and medical devices resistant to fungal colonization.
Area of Science:
- Microbiology
- Biomaterials Science
- Medical Mycology
Background:
- Candida tropicalis is a significant non-Candida albicans pathogen causing medical device-associated infections.
- Biofilm formation by C. tropicalis contributes to virulence, antifungal resistance, and medical device biofouling.
- Understanding C. tropicalis biofilm formation is crucial for developing effective prevention and treatment strategies.
Purpose of the Study:
- To optimize growth conditions for Candida tropicalis biofilm formation on polypropylene surfaces.
- To investigate the influence of various parameters on biofilm development across different clinical isolates.
- To establish a high-throughput platform for antifungal screening and medical device development.
Main Methods:
- Response surface methodology (RSM) with central composite design (CCD) was employed to optimize growth parameters.
- Parameters optimized included temperature, pH, shaker speed, inoculum size, and incubation time.
- Biofilm formation was quantified using crystal violet, MTT assay for cell viability, calcofluor white for biomass, and wet/dry weight measurements.
Main Results:
- Optimal growth conditions for C. tropicalis biofilm formation on polypropylene were successfully determined.
- Four isolates (C4, U873, U951, U1179) fit the CCD model; others required Johnson Transformation (CCD-JT) for unified conditions.
- Temperature and pH were key factors for some isolates, while temperature and incubation time were significant for others, showing strain-specific responses.
Conclusions:
- The study successfully optimized C. tropicalis biofilm growth conditions on polypropylene, identifying key influencing factors.
- A direct correlation between cell viability and biofilm formation was observed, with variations in biomass indicating strain-specific virulence.
- The developed optimization strategy serves as a valuable platform for antifungal research and designing resistant medical devices.
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