ABSTRACT Bakreshwar hot springs have geological significance, and are known to have curative properties for any diseases. In this study, antimicrobial activities of the hot spring Bakreshwar water were assessed against MDR and ATCC microorganisms. A range of bacterial strains, including Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), Escherichia coli (MDR), Klebsiella sp. (MDR), Acinetobacter sp. (MDR) and Pseudomonas sp. (MDR) were used to evaluate the antimicrobial activity of the water. The results showed that the hot spring water expressed significant antimicrobial activity against all tested MDR and ATCC microorganisms. High temperature and strong alkaline pH (9.2) of the hot spring water, the presence of minerals and the physical and chemical agents in the water are probably responsible for the antimicrobial activities that were observed. The findings lead us to believe that the hot spring water from Bakreshwar might be a source of natural antimicrobial properties, which could be utilized for the development of new drugs for treating infections caused by resistant bacteria. Further studies are required to confirm the specific mineral, chemical or physical agent that are responsible for the observed activity. Keywords: Bakreshwar’s hot spring water; antimicrobial activity; Staphylococcus aureus; Escherichia coli; Escherichia coli (MDR), Klebsiella sp.; Acinetobacter sp.; Pseudomonas sp.