2.5. Determination of Minimum Fungicide Concentration (MFC)

The minimum fungicidal concentration was performed as described by Costa-Orlandi et al. [35]. To be precise, 100 µL aliquots of the contents of the wells were removed and plated in specific media for each fungus. Candida spp. and C. neoformas were plated on Sabouraud dextrose agar (BD Difco™) and incubated at 37 °C for 24 h and 48 h, respectively. H. capsulatum was plated on BHI agar supplemented with 0.1% L-cysteine and 1% glucose and was subsequently incubated at 37 °C for 96 h; P. brasiliensis was plated on Fava-Neto medium at 37 °C for 72 h and dermatophytes on Sabouraud agar at 28 °C for 96 h. Concentrations greater than or equal to MIC₉₀ were used. The minimum fungicidal concentration is the lowest concentration of the compound or drug where the development of 99.9% of microorganisms does not occur [35].