The use of ram horn hydrolysate (RHH) as a substrate for lactic acid production was investigated using Lactobacillus casei. For this purpose, first RHH was produced. Ram horns were hydrolyzed by treating with acids (6N-H2SO4 and 6N-HCl) and neutralizing the solutions. The amounts of protein, nitrogen, ash, some minerals, total sugars, total lipids and amino acids of the RHH were determined. The effect of different concentrations (1-10% v/v) of RHH on the production of biomass, lactic acid concentration and sugar consumption was investigated, and a concentration of 6% RHH was found to be optimal. The content of lactic acid in the culture broth containing 6% RHH (44 g x l(-1)) for 26 h was 22% higher than that of the control culture broth (36 g x l(-1)). From this result, RHH was demonstrated to be a suitable supplement for lactic acid production, a use that would resolve a local environmental problem.