2.5.1. Disc Diffusion Assay

Antimicrobial susceptibility assay was carried out by disc diffusion method against methicillin-resistance *Staphylococcus aureus* (MRSA ATCC 33591, MRSA ATCC NR-46071 and MRSA ATCC 46171), Gram-negative bacteria (*Salmonella typhi* (ATCC 25241) and *Escherichia coli* (ATCC 11775)) and fungus (*Fusarium moniliforme* (MTCC 6576)). Gentamicin and Nystatin discs were used as positive control. The sterile discs (6mm, Himedia) were impregnated with 30 μL of crude extract dissolved in 0.5% DMSO. Discs impregnated with 0.5% DMSO (Qualigens Fine Chemicals Pvt. Ltd., San Diego, USA) were used as solvent control. The plates were left for 30 min at 4 ◦C to allow the diffusion of extracts before they were incubated for 24–48 h at 37 ◦C. The clear zones of inhibition observed around discs suggested antagonistic activity against test organisms and diameter of inhibition zones were measured subsequently. The test was performed in triplicate.
