Working culture was prepared with 1% inoculation level in MRS broth (50 mL) and then incubated at the same conditions for 14 h.
Duplicate Samples of each culture broth were taken every 2 h and the optical density (OD) at 560 nm was measured in a spectrophotometer (2100-UV, Unico, NJ).
A growth curve was plotted from average data as OD versus time of incubation and the time of early stage stationary phase was determined. This information was used in the subsequent studies to obtain cells at the early stationary phase for inoculation in sterile orange juice.