The purpose of the current investigation was to explore the impact of the three extenders Tris-royal jelly (TRJ), Tris-royal jelly Dimethyl Sulphoxide (TRJ DMSO) and Tris-royal jelly Ethylene Glycol (TRJ EG) on freezability of buffalo bull semen. Semen from five mature buffalo bulls was gathered. The main extender was Tris-citric acid- fructose egg yolk (TCFY) . Tris diluent having zero RJ, zero DMSO and zero EG was considered to be the control. The other extenders were Tris containing Royal Jelly TRJ(0.05%RJ), Tris containing Royal Jelly and Dimethyl Sulfoxyde TRJ DMSO(0.05%RJ+1.5%DMSO) and Tris containing Royal Jelly and Ethylene glycol TRJ EG(0.05%RJ+1.5%EG). The semen specimens were supplementary and neat concentration of spermatooa 60 × 10⁶/mL was obtained. Diluted semen was frozen. The frozen post-thawed extended buffalo semen revealed significant amelioration in all the studied parameters of the three extenders (TRJ, TRJ DMSO and TRJ EG) If compared to the control. TRJ DMSO was the best ameliorating of sperm cryosurvivability followed by TRJ and TRJ EG. Significant decrease in post-thawing sperm motility with the advance of time in all the extenders used. Significant elevation of the total antioxidants (TAC) and non- significant decrease of malondialdehyde (MDA) of the three extenders relative to the control. The superior conception rate(CR) was obvious in TRJ DMSO(64.6%) followed by TRJ(60%) and TRJ EG(57%) if compared to the control(40%). It can be fulfilled that, Royal Jelly DMSO is the best ameliorating for sperm cryosurvivability and fertility followed by Royal Jelly and Royal Jelly Ethylene glycol.