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

Tatiana Sobianski Herman, Camila da Silva Goersch, Anamelia Lorenzetti Bocca, Larissa Fernandes
Author Information
  1. Tatiana Sobianski Herman: Postgraduate Program in Molecular Pathology, Faculty of Medicine, University of Brasília, UnB, Brasília, DF, Brazil.
  2. Camila da Silva Goersch: 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.
  3. Anamelia Lorenzetti Bocca: Postgraduate Program in Molecular Pathology, Faculty of Medicine, University of Brasília, UnB, Brasília, DF, Brazil.
  4. Larissa Fernandes: 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. larissaf@unb.br. ORCID

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. 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.

Keywords

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Grants

  1. 00193-00000180/2019-91/Fundação de Apoio à Pesquisa do Distrito Federal
  2. 193.000.805/2015/Fundação de Apoio à Pesquisa do Distrito Federal
  3. Scholarship/Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

MeSH Term

Xanthenes
Oxazines
Antifungal Agents
Microbial Sensitivity Tests
Humans
Fonsecaea
Chromoblastomycosis
Colorimetry

Chemicals

resazurin
Xanthenes
Oxazines
Antifungal Agents

Word Cloud

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