Production of Lactobacillus salivarius i 24, a probiotic strain for chicken, was studied in batch fermentation using 500 mL Erlenmeyer flask. Response surface method (RSM) was used to optimize the medium for efficient cultivation of the bacterium. The factors investigated were yeast extract, glucose and initial culture pH. A polynomial regression model with cubic and quartic terms was used for the analysis of the experimental data. Estimated optimal conditions of the factors for growth of L. salivarius i 24 were; 3.32 % (w/v) glucose, 4.31 % (w/v) yeast extract and initial culture pH of 6.10.