In the present study, morphological and physiological observations were conducted to examine the effects of glycerol concentration and to determine the optimum concentration for frozen-thawed fowl spermatozoa, especially focussed on the injuries of the acrosome and midpiece of spermatozoa. The highest motility was obtained at 10% glycerol, and the secondary was at 7%. The incidence of crooked-necked spermatozoa (CNS) and acrosomal deterioration in frozen-thawed semen samples were greater in higher glycerol concentrations. Both incidence of CNS and acrosomal deterioration in frozen-thawed semen diluted with the solution containing 7% glycerol were relatively low in all the concentrations of glycerol. These results may suggest that the morphological injuries increase with the increase of glycerol concentration, and that the optimum glycerol concentration for frozenthawed fowl spermatozoa is about 7%.