Objective: To investigate the effect of extender supplementation with different concentrations of a honey bee on post-thawed sperm motility, viability index, membrane and acrosome integrities in Arab stallion. Methods: Five ejaculates from each of four Arabian stallions were subjected to cryopreservation with a modified INRA-82, without any supplementation (control) or supplemented with 1%, 2%, 3%, 4% and 5% honey bee. After thawing, all samples were maintained at 37 °C, while analyses were performed at 0, 1, and 2 and 3 h. Sperm motility percentage, viability index percentage, membrane integrity percentage and acrosome integrity percentage of each sample were determined by conventional laboratory methods. Results: Relative to the control group, supplementation with honey (2%, 3% and 4% significantly improved (P < 0.01 at least) post-thaw sperm motility, viability index (P < 0.001 at least) and had a positive effect on membrane integrity and intact acrosome percentage (P < 0.001 at least) at 0, 1, 2 and 3 h post-thawing. For all semen parameters, the lower concentration of honey (1%) and higher concentration (5%) did not show significant differences (P > 0.05) compared with the control. Conclusion: Honey bee supplementation in Arab stallion semen extenders provided a better protection of sperm parameters against cryopreservation injury, in comparison to the control groups