Kuwano river, a vital lifeline for Basti District, Uttar Pradesh, India, has faced increasing pressures due to rapid urbanization, industrialization and population growth. These anthropogenic activities have significantly impacted the river’s water quality, raising concerns about public health. To comprehensively evaluate the current state of the Kuwano River, this study employed the Most Probable Number (MPN) test to assess indicator bacteria levels in water samples collected from Ahmat Ghat in Basti City. The results revealed alarming levels of microbial contamination, rendering the river water unsuitable for both drinking and bathing. The findings from this research provide valuable baseline data for scientists, professors, policymakers and engineers to develop effective strategies for protecting and restoring the Kuwano River’s water quality. By understanding the relationship between indicator bacteria and activities, informed decisions can be made to safeguard this nationally and internally water resource.