This study aimed to evaluate the antibacterial activity of Eucalyptus, Citrus, and Sider honey against Staphylococcus aureus and Escherichia coli, to determine minimum inhibitory concentration (MIC) and synergistic effects of honey and antibiotics. Honey samples were collected directly from beekeepers. The molecularly identified Gram-negative E. coli and Gram-positive S. aureus bacterial strains used in this study were acquired from the lab of the National University of Medical Sciences (NUMS), Rawalpindi. Assessment of antimicrobial activity of honey was done by agar well diffusion method, determination of pH and electrical potential of honey, and minimum inhibitory concentration (MIC) were also done. The evaluation of the synergistic effect of various honey concentrations with the antibiotic, Augmentin (Amoxicillin/clavulanic acid 20/10µg), was performed. The study revealed that the selected honey types shown inhibition zones against both selected strains. The zone of inhibition values of gram-positive bacteria were significantly higher than those of gram-negative bacteria, indicating that the former were more vulnerable to all investigated honey types. The disc diffusion method demonstrated the remarkable synergistic effects of antibiotic (Augmentin) with all honey types against S. aureus at 50%, 75% and 100% of honey concentrations, however it did not exhibit any synergistic effects with all honey types except with Citrus honey at 75% concentration against E. coli. The conclusion is that the Sider, Citrus and Eucalyptus honey possesses remarkable antimicrobial activity against both pathogenic bacteria; S. aureus and (anti-biotic resistant) E. coli. However, the Eucalyptus honey showed the maximum antibacterial activity than the other two honey types.