Honey serves as a medicinal food that is utilized for both preventative care and the treatment of various ailments. Amidst the contemporary challenge of antibiotic resistance, honey emerges as a promising natural antimicrobial solution. The efficacy of honey in therapy hinges on its mechanisms of antimicrobial action. Thus, this study investigated the non-peroxide antibacterial properties of honey sources from a stingless bee species, Meliponula (Axestotrigona) ferruginea, that is commonly managed in Tanzania. The findings reveal that honey from stingless bees exhibits remarkable antibacterial efficacy against both resistant and susceptible bacterial strains. Notably, the studied honey samples retained a substantial portion of their antibacterial potency (89.9 - 98.7%) even after the removal of hydrogen peroxide. Interestingly, the antibacterial activity of honey did not correlate with its total phenolic and flavonoid content, suggesting the influence of specific bioactive compounds rather than overall phytochemical content. Stingless bee honey was most effective against gram-positive bacterial strains, particularly Staphylococcus aureus. These results underscore the therapeutic potential of stingless bee honey for the management of pathogenic bacteria. Future investigations should focus on elucidating the specific bioactive compounds present in stingless bee honey to bolster its clinical applications.