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Resazurin to determine the minimum inhibitory concentration on antifungal susceptibility assays for Fonsecaea sp. using a modified EUCAST protocol

  • Clinical Microbiology - Research Paper
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Abstract

Chromoblastomycosis is a fungal chronic disease, which affects humans, especially in cutaneous and subcutaneous tissues. There is no standard treatment for Chromoblastomycosis, and it is a therapeutic challenge, due natural resistance of their causative agents, inadequate response of patients and common cases of relapse. Protocols for determination of antifungal drugs susceptibility are not standardized for chromoblastomycosis agents and endpoint definition is usually based on visual inspection, which depends on the analyst, making it sometimes inaccurate. We presented a colorimetric and quantitative methodology based on resazurin reduction to resofurin to determine the metabolic status of viable cells of Fonsecaea sp. Performing antifungal susceptibility assay by a modified EUCAST protocol allied to resazurin, we validated the method to identify the minimum inhibitory concentrations of itraconazole, fluconazole, amphotericin B, and terbinafine for eight Fonsecaea clinical isolates. According to our data, resazurin is a good indicator of metabolic status of viable cells, including those exposed to antifungal drugs.

Lay abstract

This work aimed to test resazurin as an indicator of the metabolic activity of Fonsecaea species in susceptibility assays to antifungal drugs. Species of this genus are the main causative agents of Chromoblastomycosis, which affects humans.

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Acknowledgements

We thank Nucleus of Mycology from Adolfo Lutz Institute, Oswaldo Cruz Foundation (Fiocruz) and University of Paraná (Paraná Network of Biological Collections) for kindly providing Fonsecaea isolates used in this study.

Funding

The research was funded by Fundação de Apoio à Pesquisa do Distrito Federal (FAP-DF) [DEMANDA ESPONTÂNEA 00193–00000180/2019–91 to LF and FAP-DF-DEMANDA ESPONTÂNEA 193.000.805/2015 to ALB]; Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) [PhD Scholarship to TSH].

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Authors and Affiliations

  1. Postgraduate Program in Molecular Pathology, Faculty of Medicine, University of Brasília, UnB, Brasília, DF, Brazil

    Tatiana Sobianski Herman & Anamelia Lorenzetti Bocca

  2. Laboratory of Applied Immunology, Department of Cell Biology, Institute of Biological Sciences, University of Brasília, UnB, Room J1 28/8, Brasília, DF, 70910-900, Brazil

    Tatiana Sobianski Herman, Camila da Silva Goersch, Anamelia Lorenzetti Bocca & Larissa Fernandes

  3. Postgraduate Program in Microbial Biology, Institute of Biological Science, University of Brasília, UnB, Brasília, DF, Brazil

    Camila da Silva Goersch & Larissa Fernandes

  4. Faculty of Ceilândia, University of Brasília, UnB, Brasília, DF, Brazil

    Larissa Fernandes

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Herman, T.S., da Silva Goersch, C., Bocca, A.L. et al. Resazurin to determine the minimum inhibitory concentration on antifungal susceptibility assays for Fonsecaea sp. using a modified EUCAST protocol. Braz J Microbiol (2024). https://doi.org/10.1007/s42770-024-01293-2

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