SPORULATION BROTH

Description

Sporulation Broth was originally developed by Arret and Kirshbaum for the detection of antibiotic residues in milk and milk products. The medium is specified for the preparation of spores, by APHA, for disc assay procedure for the detection of sulfa drugs and antibiotics in milk. A fresh slant culture of Bacillus subtilis is washed with sterile physiological saline onto the surface of Roux bottles containing 300 ml sterile medium. The bottles are incubated at 35?C for 5 days and the resulting growth is suspended into 50 ml of sterile physiological saline. The growth is washed by centrifuging the suspension and discarding the supernatant. The sediment obtained is re-suspended in fresh sterile saline and heated at 70?C for 30 minutes to kill vegetative cells and obtain the spore suspension. This spore suspension can be stored for months for use in detection of penicillin/ antibiotic residues in milk and dairy products.