|Isolation of Bacillus: samples were suspended in saline and vortexed; supernatants were heated to 80°C for 15 minutes to kill all vegetative, non-endospore bearing cells. Supernatants were used for direct isolation on solid media. |
bSolid media (without glutamic acid supplementation) for isolation: Luria agar (pH 7–11, NaCl 0–100 g/l) and synthetic medium containing 20 g citrate/l and 40 g glycerol/l (pH 7).
cBacterial isolates were inoculated into Luria broth and incubated for 18 to 48 h. Gram staining, capsule staining (Maneval’s method), and endospore (Schaeffer and Fulton method) staining done at various intervals. All isolates showed presence of Gram positive rods in chains or single cells with capsule and endospores at different times and were further identified.