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Application of fluorescent probes to study structural changes in Aspergillus fumigatus exposed to amphotericin B, itraconazole, and voriconazole

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Abstract

The broad objective of this study was to document patterns of structural changes following antifungal treatment, and to determine any relationship with minimum inhibitory concentration (MIC) of an antifungal. Three clinical isolates of Aspergillus fumigatus, with high, intermediate, and low amphotericin B (AB), itraconazole (IZ), and voriconazole (VZ) MICs were studied in 24-well plates with cover slips. The fluorescent probes used were Calcofluor White (cell wall), propidium iodide (nucleus), and MitoTracker Green FM (mitochondria). Fluorescent microscopy as early as 3-h after exposure revealed that AB treated hyphae had intact cell wall with deformed mitochondria and nuclei while IZ and VZ treated hyphae revealed no intact cell wall, and deformation of mitochondria and nuclei. At 48 h, AB treated cells revealed rupture of hyphae and disintegration of mitochondria, and nuclei, IZ treated hyphae were swollen with disintegration of mitochondria, and nuclei while VZ treated hyphae showed rupture and disintegration of mitochondria and nuclei. The structural changes for the three strains studied were similar in fluorescent microscopy as long as the incubation time and their respective MICs were used. Thus, AB, IZ, and VZ induced gross organelle defects in A. fumigatus nuclei, mitochondria, and cell wall, which were consistent with respective MICs of antifungals used.

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Authors and Affiliations

  1. Mycology Laboratory, Wadsworth Center, NYSDOH, 120 New Scotland Ave., Albany, NY, 12208 - 2002, USA

    Madhurama Gangwar, Rama Ramani & Vishnu Chaturvedi

  2. Advanced Light Microscopy Core, New York State Department of Health, Wadsworth Center, New York, USA

    Richard Cole

  3. Department of Biomedical Sciences, School of Public Health, State University of New York at Albany, Albany, NY, USA

    Vishnu Chaturvedi

  4. Pfizer Inc., New York, NY, USA

    Daniel J. Sheehan

  5. Department of Microbiology, Punjab Agricultural University, Ludhiana, India

    Madhurama Gangwar

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  1. Madhurama Gangwar
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Correspondence to Vishnu Chaturvedi.

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Gangwar, M., Cole, R., Ramani, R. et al. Application of fluorescent probes to study structural changes in Aspergillus fumigatus exposed to amphotericin B, itraconazole, and voriconazole. Mycopathologia 162, 103–109 (2006). https://doi.org/10.1007/s11046-006-0040-y

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  • DOI: https://doi.org/10.1007/s11046-006-0040-y

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