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Tea tree oil is a yellow liquid extracted from Melaleuca alternifolia. It has an excellent antibacterial, antifungal, anti-inflammatory activity, which is used topically in niosomal gel for bacterial infection. Tea Tree oil-loaded noisome was prepared by thin-film hydration technique. Nine different batches are prepared by changing cholesterol: span 60 ratios, hydrating speed, and hydrating time. All batches were evaluated for entrapment efficiency; batch F8 had the highest entrapment efficiency of 73%. An optimized batch was selected to prepare niosomal gel by dispersion method using carbopol, propylene glycol. Different batches of the gel prepared by changing the concentration of carbopol and formulated gel were evaluated for various parameters like homogeneity, grittiness, pH, drug content, spreadability, In-vitro drug release. The cup plate method was used to check the antibacterial activity against different microorganisms. The drug and formulation were tested against Staphylococcus aureus and Candida albicans. Out of all batches, TTNG6 was found more effective with drug content 75.34%, Spreadability 58.12, pH 7.01, viscosity 10324 cps. Tea tree oil optimized niosomal gel was found more stable with good antibacterial properties than conventional gel.