2.4. Live culture maintenance
As a control, cultures (100 mL) were maintained by weekly subculturing.
Flasks containing 90 mL of fresh growth medium (MBSM) were inoculated with 10 mL of the previous culture (10% v/v) and incubated at temperatures specified in Table 1 for 5 weeks, with the subculturing process repeated every 7 days.
Bioleaching tests were carried out to determine leaching activity of the live cultures in duplicate after a total of 5 weeks of weekly subculturing.
Cell survival was determined after 1 week and 5 weeks of weekly subculturing as described previously.