Antibiotic resistance in coliform organisms isolated from the Ganga River waters across its Upper, Middle, and Lower stretches was assessed. The study revealed varying levels of antibiotic resistance among the coliform isolates, with differences observed across the different stretches of the river. In the Upper stretch, sites like Devprayag and Gangotri exhibited high levels of E. coli contamination, with corresponding moderate antibiotic resistance (MAR) values. In the Middle stretch, sites like Farrukhabad and Bithoor showed lower E. coli contamination compared to the Upper stretch but had higher MAR values. The Lower stretch, particularly sites like Revelganj and Bahachoki in Bihar, displayed significantly elevated E. coli contamination levels and MAR values. These findings underscore the presence of antibiotic-resistant coliform organisms in different segments of the Ganga River, emphasizing the need for targeted interventions to mitigate antibiotic resistance spread in this vital water resource. Understanding the antibiotic resistance profiles at specific sites along the Ganga River can inform strategies to preserve water quality and protect public health in regions dependent on this iconic river.