Artificial insemination (AI) is the best to tool to improve the reproductive and productive performance of large/ small ruminants. Semen cryopreservation, an important step in AI, may result into damages to spermatozoa due to oxidation process. Different antioxidants have been used successfully in the extenders of buck semen. Royal jelly is also reported to be a good antioxidant for ram and buffalo bull semen. No prior study is available on the use of royal jelly in the extenders of buck semen. The objectives of this study were to evaluate and optimize the different concentration of royal jelly in semen extender and its effect on post thaw quality of Beetal buck sperm. A total of 20 one ejaculates were collected from 3 mature Beetal bucks during 7 successive weeks. At each collection, the ejaculates from 3 bucks passing the minimum criteria (with motility ≥60%, concentration >2 × 10⁹ sperm/mL and abnormality rate < 10%) were pooled and divided into 5 aliquots. The aliquots were extended (60 × 10⁶/0.5mL straw) at 37°C with Tris-Citric acid Egg Yolk (TCEY) extender with different concentrations of Royal Jelly (0, 0.5, 1.0, 1.5 or 2.0 mM). The extended semen samples were cooled to 4°C for 90 min, equilibrated at 4°C for 2 h, frozen in liquid nitrogen (LN₂) vapor (4 cm above the LN₂, for 7 min) and plunged into LN₂ for storage. Post thaw semen evaluation assays, e.g., motility, plasma membrane integrity (PMI) using HOST, normal apical ridge (NAR) and sperm viability using FITC-PNA/PI and DNA integrity using acridine orange were evaluated using phase contrast and fluorescent microscopy. Data were analyzed through ANOVA under CRD and significant differences (P < 0.05%) were compared by using Duncan’s Multiple Range Test. Royal Jelly, at an inclusion level of 1.0 mM significantly (P < 0.05) improved post thaw motility, PMI, NAR, live percentage and DNA integrity as compared with control and other treatment groups. It is concluded that 1.0 mM concentration of royal jelly in TCEY semen extender may prove beneficial for buck semen.