Three experiments were carried out to find the optimal concentration of DMSO and glycerol in BFS diluent for freezing rabbit spermatozoa. Semen was diluted 1:1 with diluent A (BF5 + DMSO) at 25°C and diluted further 1:1 with diluent B (diluent A + glycerol) after cooling down to 5°C. Diluted semen was frozen immediately and stored in liquid nitrogen. Maximum percentages for motility and normal acrosomes were obtained in the presence of 12% DMSO (as expressed in diluent A) and 3% glycerol (final concentration) after thawing.