Mycoplasma agalactia was cultured in PPLO broth medium supplemented with sterilized normal horse serum in fermentor. Cultivation process continued at 37 ºC under aerobic conditions and growth was terminated after 26 hours cultured period with 2.2×109 Mycoplasma per ml. Samples were taken at different stages of growth. Culture was inactivated with 0.04 %v/v formaldehyde and the vaccine was adjuvanted with saponin with final concentration of 1/1000 w/v. Results indicated that the yield of Mycoplasma was higher than the vaccine produced in large flasks using the same culture media. The produced vaccine in the fermentor was safe in guinea pig, sheep and goat according to OIE instructions. Based on the present findings, Mycoplasma agalatia can easily grow in fermentor for the purpose of vaccine production instead of its conventional production in large flasks.