Bacterial Growth and Incubation Conditions With Additional of 5% NaCl
Overnight cultures of S. aureus were grown in Tryptic Soy Broth (TSB) at 37°C with constant agitation (120 rpm). Eight flasks containing 95 ml TSB culture media were inoculated with 5 ml of overnight culture in 500 ml conical flasks which were incubated at 37°C with constant agitation (120 rpm) for 3 h. Replicate cultures were harvested and washed three times using phosphate buffer saline (PBS) to ensure the elimination of TSB medium. Washed cells were then incubated in a defined minimal medium consisting of PBS with trace as described in Alreshidi et al. (2019). The incubation medium did not allow the cells replication but provided required elements and nutrients bases of energy and nitrogen that help bacterial cells to have active metabolism. The number of cells was assessed by the plating techniques to determine colony forming units (CFU) at initial and final times of incubation for both control and NaCl stressed samples.