Biofilm characterization of Fusarium solani keratitis isolate: increased resistance to antifungals and UV light

Itzel Margarita Córdova-Alcántara; Diana Laura Venegas-Cortés; María Ángeles Martínez-Rivera; Néstor Octavio Pérez; Aida Verónica Rodriguez-Tovar

doi:10.1007/s12275-019-8637-2

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Biofilm characterization of Fusarium solani keratitis isolate: increased resistance to antifungals and UV light: Itzel Margarita Córdova-Alcántara ¹, Diana Laura Venegas-Cortés ¹, María Ángeles Martínez-Rivera ¹, Néstor Octavio Pérez ², Aida Verónica Rodriguez-Tovar ¹; Journal of Microbiology 2019;57(6):485-497.
DOI: https://doi.org/10.1007/s12275-019-8637-2
Published online: May 27, 2019

¹Medical Micology Laboratory, Microbiology Department National School of Biological Sciences, National Polytechnique Institute (IPN), C.P. 11340 Ciudad de México, México, ²Research and Development Department, Probiomed S.A. de C.V. Cruce de Carreteras Acatzingo-Zumpahuacan S/N C.P. 52400 Tenancingo, Estado de México, México¹Medical Micology Laboratory, Microbiology Department National School of Biological Sciences, National Polytechnique Institute (IPN), C.P. 11340 Ciudad de México, México, ²Research and Development Department, Probiomed S.A. de C.V. Cruce de Carreteras Acatzingo-Zumpahuacan S/N C.P. 52400 Tenancingo, Estado de México, México

Corresponding author: Néstor Octavio Pérez , Tel: +52-55-11662305,
Aida Verónica Rodriguez-Tovar , Tel: +52-55-11662305,

Received: 21 November 2018 • Revised: 23 January 2019 • Accepted: 28 January 2019

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Abstract

Fusarium solani has drawn phytopathogenic, biotechnological, and medical interest. In humans, it is associated with localized infections, such as onychomycosis and keratomycosis, as well as invasive infections in immunocompromised patients. One pathogenicity factor of filamentous fungi is biofilm formation. There is still only scarce information about the in vitro mechanism of the formation and composition of F. solani biofilm. In this work, we describe the biofilm formed by a clinical keratomycosis isolate in terms of its development, composition and susceptibility to different antifungals and ultraviolet light (UV) at different biofilm formation stages. We found five biofilm formation stages using scanning electron microscopy: adherence, germination, hyphal development, maturation, and cell detachment. Using epifluorescence microscopy with specific fluorochromes, it was elucidated that the extracellular matrix consists of carbohydrates, proteins, and extracellular DNA. Specific inhibitors for these molecules showed significant biofilm reductions. The antifungal susceptibility against natamycin, voriconazole, caspofungin, and amphotericin B was evaluated by metabolic activity and crystal violet assay, with the F. solani biofilm preformation to 24 h increased in resistance to natamycin, voriconazole, and caspofungin, while the biofilm preformation to 48 h increased in resistance to amphotericin B. The preformed biofilm at 24 h protected and reduced UV light mortality. F. solani isolate could produce a highly structured extra biofilm; its cellular matrix consists of carbohydrate polymers, proteins, and eDNA. Biofilm confers antifungal resistance and decreases its susceptibility to UV light. The fungal biofilm functions as a survival strategy against antifungals and environmental factors.

Keywords: Fusarium solani;biofilm;scanning electron microscopy;antifungal susceptibility;epifluorescence microscopy;extracellular matrix;ultraviolet light

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Biofilm characterization of Fusarium solani keratitis isolate: increased resistance to antifungals and UV light

J. Microbiol. 2019;57(6):485-497. Published online May 27, 2019

DOI: https://doi.org/10.1007/s12275-019-8637-2

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