Biofilm development of <i>Candida boidinii</i> and the effect of tyrosol on biofilm formation

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Abstract

Objectives

The applicability of a simple and high-throughput method for quantitative characterization of biofilm formation by Candida boidinii was tested in order to evaluate the effects of exogenous tyrosol on yeast growth and biofilm formation capacity.

Results

Significant concentration-, temperature and time-dependent effect of tyrosol (2-(4-hydroxyphenyl)ethanol) was demonstrated, but it differentially affected the growth and biofilm formation (characterized by crystal violet staining and XTT-reduction assay) of Candida boidinii. Testing biofilm based on metabolic activity displayed sensitively the differences in the intensity of biofilm in terms of temperature, tyrosol concentration, and exposure time. At 22 °C after 24 h none of the tyrosol concentrations had significant effect, while at 30 °C tyrosol-mediated inhibition was observed at 50 mM and 100 mM concentration. After 48 h and 72 h at 22 °C, biofilm formation was stimulated at 6.25–25 mM concentrations, meanwhile at 30 °C tyrosol decreased the biofilm metabolic activity proportionally with the concentration.

Conclusions

The research concludes that exogenous tyrosol exerts unusual effects on Candida boidinii growth and biofilm formation ability and predicts its potential application as a regulating factor of various fermentations by Candida boidinii.

Supplementary Information

The online version contains supplementary material available at 10.1007/s10529-023-03432-5.

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Most cited references 39

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Microtiter dish biofilm formation assay.

George A O'Toole (2011)

Biofilms are communities of microbes attached to surfaces, which can be found in medical, industrial and natural settings. In fact, life in a biofilm probably represents the predominate mode of growth for microbes in most environments. Mature biofilms have a few distinct characteristics. Biofilm microbes are typically surrounded by an extracellular matrix that provides structure and protection to the community. Microbes growing in a biofilm also have a characteristic architecture generally comprised of macrocolonies (containing thousands of cells) surrounded by fluid-filled channels. Biofilm-grown microbes are also notorious for their resistance to a range of antimicrobial agents including clinically relevant antibiotics. The microtiter dish assay is an important tool for the study of the early stages in biofilm formation, and has been applied primarily for the study of bacterial biofilms, although this assay has also been used to study fungal biofilm formation. Because this assay uses static, batch-growth conditions, it does not allow for the formation of the mature biofilms typically associated with flow cell systems. However, the assay has been effective at identifying many factors required for initiation of biofilm formation (i.e, flagella, pili, adhesins, enzymes involved in cyclic-di-GMP binding and metabolism) and well as genes involved in extracellular polysaccharide production. Furthermore, published work indicates that biofilms grown in microtiter dishes do develop some properties of mature biofilms, such a antibiotic tolerance and resistance to immune system effectors. This simple microtiter dish assay allows for the formation of a biofilm on the wall and/or bottom of a microtiter dish. The high throughput nature of the assay makes it useful for genetic screens, as well as testing biofilm formation by multiple strains under various growth conditions. Variants of this assay have been used to assess early biofilm formation for a wide variety of microbes, including but not limited to, pseudomonads, Vibrio cholerae, Escherichia coli, staphylococci, enterococci, mycobacteria and fungi. In the protocol described here, we will focus on the use of this assay to study biofilm formation by the model organism Pseudomonas aeruginosa. In this assay, the extent of biofilm formation is measured using the dye crystal violet (CV). However, a number of other colorimetric and metabolic stains have been reported for the quantification of biofilm formation using the microtiter plate assay. The ease, low cost and flexibility of the microtiter plate assay has made it a critical tool for the study of biofilms.

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Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates.

Elke Peeters, Hans Nelis, Tom Coenye (2008)

In the present study six assays for the quantification of biofilms formed in 96-well microtiter plates were optimised and evaluated: the crystal violet (CV) assay, the Syto9 assay, the fluorescein diacetate (FDA) assay, the resazurin assay, the XTT assay and the dimethyl methylene blue (DMMB) assay. Pseudomonas aeruginosa, Burkholderia cenocepacia, Staphylococcus aureus, Propionibacterium acnes and Candida albicans were used as test organisms. In general, these assays showed a broad applicability and a high repeatability for most isolates. In addition, the estimated numbers of CFUs present in the biofilms show limited variations between the different assays. Nevertheless, our data show that some assays are less suitable for the quantification of biofilms of particular isolates (e.g. the CV assay for P. aeruginosa).

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Critical review on biofilm methods.

Joana Azeredo, Nuno F Azevedo, Romain Briandet … (2017)

Biofilms are widespread in nature and constitute an important strategy implemented by microorganisms to survive in sometimes harsh environmental conditions. They can be beneficial or have a negative impact particularly when formed in industrial settings or on medical devices. As such, research into the formation and elimination of biofilms is important for many disciplines. Several new methodologies have been recently developed for, or adapted to, biofilm studies that have contributed to deeper knowledge on biofilm physiology, structure and composition. In this review, traditional and cutting-edge methods to study biofilm biomass, viability, structure, composition and physiology are addressed. Moreover, as there is a lack of consensus among the diversity of techniques used to grow and study biofilms. This review intends to remedy this, by giving a critical perspective, highlighting the advantages and limitations of several methods. Accordingly, this review aims at helping scientists in finding the most appropriate and up-to-date methods to study their biofilms.

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Author and article information

Contributors

Mónika Molnár:

ORCID: http://orcid.org/0000-0001-5296-7924

molnar.monika@vbk.bme.hu

Journal

Journal ID (nlm-ta): Biotechnol Lett

Journal ID (iso-abbrev): Biotechnol Lett

Title: Biotechnology Letters

Publisher: Springer Netherlands (Dordrecht )

ISSN (Print): 0141-5492

ISSN (Electronic): 1573-6776

Publication date (Electronic): 13 October 2023

Publication date PMC-release: 13 October 2023

Publication date (Print): 2023

Volume: 45

Issue: 11-12

Pages: 1541-1554

Affiliations

Budapest University of Technology and Economics Department of Applied Biotechnology and Food Science, ( https://ror.org/02w42ss30) Műegyetem rkp. 3, 1111 Budapest, Hungary

Author information

Mónika Molnár http://orcid.org/0000-0001-5296-7924

Article

Publisher ID: 3432

DOI: 10.1007/s10529-023-03432-5

PMC ID: 10635961

PubMed ID: 37831285

SO-VID: 4f7a3719-55f5-4a6a-aca4-c75904912897

License:

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

History

Date received : 29 November 2022

Date revision received : 26 July 2023

Date accepted : 6 September 2023

Funding

Funded by: Richter Gedeon Talentum Foundation

Award ID: Richter Gedeon Talentum Foundation

Award Recipient : Rita Márton

Funded by: FundRef http://dx.doi.org/10.13039/501100012550, Nemzeti Kutatási, Fejlesztési és Innovaciós Alap;

Award ID: TKP2021-EGA-02

Award Recipient : Mónika Molnár

Funded by: Budapest University of Technology and Economics

Open Access :

Open access funding provided by Budapest University of Technology and Economics.

Custom metadata

ScienceOpen disciplines: Biotechnology

Keywords: biofilm formation,candida boidinii,quorum sensing,tyrosol

Data availability:

ScienceOpen disciplines: Biotechnology

Keywords: biofilm formation, candida boidinii, quorum sensing, tyrosol

Biofilm development of Candida boidinii and the effect of tyrosol on biofilm formation

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Abstract

Objectives

Results

Conclusions

Supplementary Information

Related collections

Bacterial vaginosis and biofilm

Most cited references 39

Microtiter dish biofilm formation assay.

Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates.

Critical review on biofilm methods.

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Contributors

Journal

Affiliations

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History

Funding

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