Conventional methods for the propagation and preservation of parasites in vivo or in vitro have limitations, which include the need for labor, the initial isolation and loss of strains, bacterial and fungal contamination, and changes in the original biological and metabolic characteristics of the organism. All of these disadvantages are reduced considerably by cryopreservation. In this study, we examined the effects of different concentrations of dimethyl sulfoxide (DMSO) and glycerol on the survival rate of Trichomonasgallinae. Both of the two cryoprotectants examined at different concentrations showed a protective effect of T. gallinae. When DMSO was used as the cryoprotectant, a maximum survival rate (70%) was achieved at a concentration of 10% over a short-term duration of freezing (24 h). Glycerol at a concentration of 10% had the highest effect on cryoprotectivity after 70 days, while DMSO 20% showed the lowest cryopreservation for T. gallinae. In conclusion, DMSO and glycerol are appropriate protective materials for the cryopreservation of T. gallinae. The solutions of DMSO 10% and glycerol 10% could be the best choice of cryoprotectant for short-term (24 hours) and long-term (70 days) protection, respectively.