Barley seeds were cleaned thoroughly and half of the raw seeds were ground in an electric grinding mill using a 1.5 mm sieve size and the rest of the seeds were soaked in distilled water for 12 h at room temperature. A seed to water ratio of 1:5 (w/v) was used. The unimbibed water was discarded. The soaked seeds were germinated in sterile petri dishes lined with wet filter paper for 24 h at 37 °C with frequent spraying of water. After 24 h, the sprouts were rinsed in distilled water and then dried at 55–60 °C. The dried samples of germinated seeds were ground to fine powder in an electric grinder and then stored in plastic containers for further use.
Two types of food mixtures were formulated from non-germinated and germinated barley flour along with whey powder and tomato pulp in the ratio of 2:1:1 w/w. Addition of tomato pulp and whey powder in food mixtures not only added nutrients, but also provided an optimum medium for growth of L. acidophilus.