Add all ingredients except Mg SO4 to 1 liter distilled water. Heat to boiling with stirring. Add Mg SO4 and adjust pH. Dispense 8 ml portions into 16 x 150 mm tubes. Autoclave 15 min at 121°C. Incline tubes to obtain 5 cm slant. Final pH, 6.7
M3. Acid Broth Proteose peptone 5 g Yeast extract 5 g Dextrose 5 g K2HPO4 4 g Distilled water l liter
Dissolve ingredients and dispense 12-15 ml portions into 20 x 150 mm tubes. Autoclave 15 min at 121°C. Final pH, 5.0.
M4. AE Sporulation Medium, Modified (for C. perfringens) (AE base is available commercially) Polypeptone 10.0 g Yeast extract 10.0 g Na2HPO4 4.36 g KH2PO4 0.25 g Ammonium acetate1.5 g Mg SO4·7H2O 0.2 g Distilled water 1 liter
Dissolve ingredients and adjust to pH 7.5 ± 0.1, using 2 M sodium carbonate. Dispense 15 ml into 20 x 150-mm screw-cap tubes and sterilize by autoclaving for 15 min at 121°C. After sterilization, add