2.5.1. Disc Diffusion Method

Antimicrobial activity of active fraction was assessed by disk diffusion method against *Staphylococcus epidermidis* (MTCC 435), *Staphylococcus aureus* (MTCC 740), *Bacillus cereus* (MTCC 1272), *Escherichia coli* (MTCC 40), *Klebsiella pneumoniae* (MTCC 661), *Micrococcus luteus* (MTCC 7950) *Aspergillus flavus* (MTCC 2590), *Fusarium moniliforme* (MTCC 6576), *Bipolaris maydis* and *Alternaria alternata* (MTCC 1362). The sterile discs (6 mm, Himedia) were impregnated with 30 μL of crude extract. The pathogens were inoculated in Mueller-Hinton broth (24 h for bacteria, Himedia) and Sabouraud Dextrose both (72 h for fungi, Himedia). The well-grown bacterial and fungal cultures were plated on Mueller-Hinton agar and Potato Dextrose agar respectively (Himedia). Sterile discs loaded with extract were placed on the plate. Chloramphenicol discs (Himedia) were used as positive control for antibacterial assay, while nystatin discs (Himedia) were used for the antifungal assay. Discs impregnated with dimethyl sulfoxide (DMSO, Himedia) were used as the solvent control. The plates were incubated at 37 ◦C and room temperature (for test bacteria and fungi respectively) and the zone of inhibition was measured.
