OBJECTIVES: The purpose of this experiment was to evaluate the antibacterial efficacy and safety profile of sequential extracts of an indigenous plant Gymnema sylvestre R.Br. (Gurmar) leaves. METHODS: This study was conducted against 4 bacteria including Staphylococcus aureus, Salmonella enterica, Clostridium perfringens type-A, Haemophillus paragallinarum which are common causatives of various infections. In order to assess antibacterial potential of Chloroform, Hexane, Ethanolic and Aqueous extracts of Gymnema sylvestre R.Br. leaves, agar well diffusion method was used. RESULTS: The results exhibited that ethanolic and chloroformic extracts have more antibacterial activity against all four microorganisms under study. Hexane, chloroform and ethanolic extracts of G. sylvestre R. Br. showed antibacterial activity against all isolates of selected 4 microorganisms (Salmonella enterica, Staphylococcus aureus, Haemophillus paragallinarum and Clostridium perfringens type-A) under study. On the other hand aqueous extract showed antibacterial activity only against Clostridium perfringens type-A while no antibacterial activity was observed against remaining three bacteria under study. MTT Assay was performed using vero cell lines. The results showed that 2900µg/ml, 3612.50µg/ml, 4075µg/ml and 1562.50µg/ml concentrations of Hexane, Chloroform, Ethanol and Aqueous extracts respectively were found safe as Cell Survival Percentage (CSP) at these concentrations was above 50%. CONCLUSIONS: The antibacterial and MTT Assay results suggest that sequential extracts of dried leaves of Gymnema sylvestre R. Br. can be used as a safer antibacterial against tested resistant bacteria. All results were compared statistically using DMR (Duncan’s Multiple Range) Post hoc test at P ≤ 0.05.