After 24 h of incubation at 42uC, an isolated black colony was selected, picked, and transferred to 9 ml of BPW in a dilution tube with cap and then incubated at 22uC for 72 h. Aliquots (0.1 ml) of the 72-h culture were then transferred to storage vials that contained 0.9 ml of brain heart infusion broth with 15% (vol/vol) glycerol.