Determination of minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC)

Minimal inhibitory concentration (MIC) of the most effective extract and fluconazole against the four selected strains was performed using 96-well microtiter plates. Selected strains were added in SDB supplemented with varying concentrations of acetone extract of C. spongiosus (AECS) and fluconazole, then incubated at 37 °C for 48 h. After incubation, the fungal growth was assayed at 600 nm using a Biotek plate reader. The MIC was recorded as the lowest concentration that produced complete suppression of visible growth⁴⁵.

The MFC of AECS and fluconazole was determined according to Borman⁴⁵. Briefly, (10 µg/ml) from MIC to last concentration wells of AECS and fluconazole were transferred separately to SDA plates, which were then incubated at 37 °C for 48 h. The MFC was recorded as the lowest drug concentration at which fungal growth was completely inhibited after 48 h of incubation.