Standardized Method for In Vitro Antifungal Susceptibility Testing of Candida albicans Biofilms
Open accessAbstract
Candida albicans is implicated in many biomaterial-related infections. Typically, these infections are associated with biofilm formation. Cells in biofilms display phenotypic traits that are dramatically different from those of their free-floating planktonic counterparts and are notoriously resistant to antimicrobial agents. Consequently, biofilm-related infections are inherently difficult to treat and to fully eradicate with normal treatment regimens. Here, we report a rapid and highly reproducible microtiter-based colorimetric assay for the susceptibility testing of fungal biofilms, based on the measurement of metabolic activities of the sessile cells by using a formazan salt reduction assay. The assay was used for in vitro antifungal susceptibility testing of several C. albicans strains grown as biofilms against amphotericin B and fluconazole and the increased resistance of C. albicans biofilms against these antifungal agents was demonstrated. Because of its simplicity, compatibility with a widely available 96-well microplate platform, high throughput, and automation potential, we believe this assay represents a promising tool for the standardization of in vitro antifungal susceptibility testing of fungal biofilms.
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Sessile Candida cells show high levels of tolerance to antifungal therapy as compared to their planktonic counterparts.
“sessile cells show high levels of tolerance to antifungal therapy as compared to their planktonic counterparts [21,22].”
From Antifungal therapy of Candida biofilms: Past, present and future (2023) · cites this paper as [22]
Cells within Candida biofilms show increased resistance against polyenes.
“From the very early reports on biofilm activity, it was already demonstrated that cells within Candida biofilms show increased resistance against polyenes [22].”
From Antifungal therapy of Candida biofilms: Past, present and future (2023) · cites this paper as [22]
Amphotericin B was approximately 10 times less potent under biofilm versus planktonic growing conditions, but the high concentrations needed to effectively kill biofilms are considered toxic and unsafe, restricting its use.
“For example, amphotericin B was approximately 10 times less potent when tested under biofilm-versus planktonic-growing conditions, but these high concentrations needed to effectively kill biofilms are considered toxic and unsafe, effectively restricting its use [22].”
From Antifungal therapy of Candida biofilms: Past, present and future (2023) · cites this paper as [22]
Biofilms needed about 10 to 100 times more antifungal drug to be eradicated compared with planktonic cells.
“These results are consistent with reports for C. albicans biofilms (Baillie and Douglas, 1998; Chandra et al., 2001a,b; Ramage et al., 2001), showing that biofilms needed about 10 to 100 times more quantity of antifungal drug to be eradicated when compared to planktonic cells.”
From Candida glabrata susceptibility to antifungals and phagocytosis is modulated by acetate (2015) · cites this paper as Ramage et al., 2001
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- Authors
- Ramage, G., Vande Walle, K., Wickes, B. L., & López-Ribot, J. L.
- Affiliations
- The University of Texas at San Antonio Health Science Center (US)
- <!--label omitted: 2-->Department of Medicine, Division of Infectious Diseases,2The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78245
- <!--label omitted: 1-->Department of Microbiology1 and
- Venue
- Antimicrobial Agents and Chemotherapy
- Publisher
- American Society for Microbiology
- Published
- 2001-09
- Volume
- 45
- Issue
- 9
- Pages
- 2475-2479
- ISSN
- 0066-4804 (ISSN-L)
- 1098-6596
- PMC
- PMC90680
- OpenAlex
- W2135536120
- Cited by
- 753
- References
- 1
- Citations by year
- 2026: 25 · 2025: 23 · 2024: 21 · 2023: 38 · 2022: 32 · 2021: 40 · 2020: 30 · 2019: 37 · 2018: 31 · 2017: 63 · 2016: 30 · 2015: 40 · 2014: 51 · 2013: 45 · 2012: 31
- FWCI
- 10.17
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Antimicrobial Agents and Chemotherapy (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- ENLIGHTEN (Jurnal Bimbingan dan Konseling Islam) (submittedVersion)
- Funders
- Burroughs Wellcome Fund
- Topics
- Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bacterial biofilms and quorum sensing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Keywords
- Candida albicans, fungal biofilms, antifungal susceptibility testing, colorimetric assay, microplates, amphotericin B, fluconazole, antifungal resistance
- MeSH terms
- Amphotericin B, Antifungal Agents, Candida albicans, Cell Count, Colorimetry, Humans, Microbial Sensitivity Tests, Reproducibility of Results, Fluconazole, Biofilms
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