In the Indian state of Uttar Pradesh, the Gomti River receives tributaries from the Sai River (Sai Setu), also known as the Aadi Ganga. The river Sai originates in the hamlet of Bijgwan, close to Pihani in the district of Hardoi. It flows for around 600 kilometres, forming the district border between Lucknow and Unnao. It then travels through the districts of Jaunpur, Raebareli, and Hardoi until joining the Gomati River in Rajepur (25°39'8.63"N 82°48'5.00"E) in Jaunpur district of Uttar Pradesh, India. The overall water quality index of the samples was ascertained by microbiological tests of twelve water samples collected from the Raebareli area: Rajghat Raebareli, Shaheed Smarak Raebareli, Behta Bridge Raebareli, and Bhanvareshvar Temple Raebareli. Here, the important and observational method is called MPN (Most Probable Number). The present study will center on the quantity of Total Coliform (TC), Faecal Coliform (FC), and Faecal Streptococci found in the Holy Water Sai. Additionally, a thorough discussion of water quality, antibiotic resistance, and sensitivity will be included. Since water is essential to all living things, including people, it must be pure and of high quality for human health, particularly in light of microbial contamination.