This study was conducted to evaluate the effect of replacing glycerol with erythritol on cryopreservation of ram spermatozoa. Semen samples (n=24) were collected from four rams in six times. In each session, the collected ejaculates (n=4) were pooled and split into 12 equal parts. The amount of 0. 032 M glycerol (G32E0, equal to 3% glycerol), 0. 016 M glycerol and 0. 016 M erythritol (G16E16), 0. 008 M glycerol and 0. 024 M erythritol (G8E24), 0. 032 M erythritol (G0E32), 0. 054 M glycerol (G54E0, equal to 5% glycerol), 0. 027 M glycerol and 0. 027 M erythritol (G27E27), 0. 013 M glycerol and 0. 041 M erythritol (G13E41), 0. 054 M erythritol (G0E54), 0. 076 M glycerol (G76E0, equal to 7% glycerol ), 0. 038 M glycerol and 0. 038 M erythritol (G38E38), 0. 019 M glycerol and 0. 057 M erythritol (G19E57) and 0. 076 M erythritol (G0E76) were added. The diluted samples were frozen using standard protocol. After thawing, the samples were incubated at 37° C for 6 h. Results showed that progressive sperm motility and acrosome integrity were higher in G13E41 (18. 85 % and 27. 41 %, respectively) than treatments that contained only glycerol at 6 h (p < 0. 05). At the level of 0. 032 and 0. 054 M cryoprotectant, the highest of total sperm motility was observed in G8E24 (19. 16 %) and G13E41 (18. 85 %) at 6 h, respectively (p < 0. 05). Therefore, the quality of frozen-thawed ram spermatozoa can be improved by using the mixture of 0. 013 M glycerol plus 0. 041 M erythritol or 0. 008 M glycerol plus 0. 024 M erythritol...