B. indicus HU36 was supplied from EU 7th Framework acronymed Project “COLORSPORE” (Project number: 207948). B. indicus HU36 spores were obtained by exhaustion method described by Nicholson and Setlow (1990). According to the method, Difco Sporulation Medium (DSM) was streaked with colony of B. indicus HU 36 and incubated overnight at 37 C. Then, a fresh colony was inoculated into Luria Bertoni (LB) Broth and incubated at 30 C on a rolling drum overnight. DSM agar and LB broth were supplied from Oxoid (Basingstoke, UK). Afterward, the culture was grown in an orbital shaking water bath at 37 C until OD600 of the culture was reached approximately 1.00. Then culture was spread plated on DSM agar and incubated at 30 C for 24 h. Afterward, the level of visible sporulation of the culture was checked frequently. When
more than 90% of visible sporulation was detected, spores were
harvested with water after removal of vegetative cells by washing
with distilled water. Finally, harvested spores were lyophilized by
using Christ Alpha 1-2 LD Plus Freeze Dryer (Martin Christ GMBH,
Osterede am Harz, Germany).