The key role of a cryoprotectant is to minimize the chemical and physical stress which occurs during cooling, freezing and thawing of semen. The difference between the cryoprotectant (CPA) occurs in their permeability coefficient and the structural model of the cryogenic agent. The beneficial effect of dimethylacetamide (DMA) as a cryoprotectant especially for sperms had been observed in several studies. The aim of the study was to study the cryoprotective effect of DMA in freezing the Malabari buck semen compared to glycerol. Ten ejaculates were taken from fourMalabaribucks . After preliminary evaluation sample split technique was followed with Tris based extender containing glycerol (6.7 per cent) as cryoprotectant (control) and Trisextender containing DMA (3 per cent) as cryoprotectant (treatment group). The semen straws (0.25mL) after filling were subjected for equilibration and manual freezing. Sperm kinetics was studied using computer-aided sperm analyzer. Pre-freeze and post-thaw evaluation included sperm viability, sperm abnormality, hypo osmotic test, acrosome integrity test and DNA fragmentation. Results indicated that inclusion of 6.7 per cent glycerol had significantly higher (p<0.05) post-thaw values than DMA. From our study we conclude that 6.7 per cent glycerol was better than 3 per cent DMA in cryopreservation of Malabari buck semen.