Evaluation of antifungal activity-agar diffusion method

The antifungal activity against the fungal strains was done according to the CLSI guidelines M44 A, 2008.[20] Wells of standard diameter (5 mm) were bored in the modified MHA using sterile well puncher. A lawn culture was done with inoculum already adjusted to 0.5 McFarland standard (1.5 × 10⁸ colony-forming unit/mL). Ten microliters, 20 μL, and 50 μL of Ag-NPs were added to the respective wells. The concentration of C-Ag-NPs in each well was 10 μg, 20 μg, and 50 μg, respectively. Fifty microliters of curcumin and 0.1 M AgNO₃ were included as controls. The concentration of curcumin in 50 μL of curcumin solution was 250 μg. The plates were then incubated at 37°C for approximately 18–24 h. The diameter of zones of inhibition produced by varied concentrations of C-Ag-NPs against the test organisms was measured after overnight incubation. Next day, the diameter of zones of inhibition produced by varied concentrations of the extracts against the test organisms was measured.