Madhuca longifolia, commonly known as Mahua, is a revered tree indigenous to the Indian subcontinent, celebrated for its versatile uses in traditional medicine and cultural practices. In this study, the antibiotic effects of ethanolic extract of Madhuca longifolia leaves was evaluated against six MDR and ATCC bacterial strains. The results showed that the leaf extract of Madhuca longifolia exhibited significant antimicrobial activity against all tested MDR bacterial strains. The plant has enormous bioactive compounds, which can be potentially responsible for the observed antimicrobial activities. Also, the synergistic effect of the leaf extract gave promising results when applied along with Amikacin and Ceftazidime-Avibactam. The acquired results suggest that Madhuca longifolia may be a potential source of natural antibacterial agents that can be used in the development of new drugs to treat resistant bacterial infections. Further studies are required to identify and isolate the active ingredients responsible for the observed activity and to evaluate their efficacy and safety in vivo.