Microbial activity in water can lead to various challenges, including the contamination of drinking water, deterioration of overall water quality, and the proliferation of harmful pathogens. In Egypt, this issue is particularly significant due to the country's heavy reliance on the Nile River, which is potentially contaminated with bacteria, parasites, and other pollutants. In the present study, thirty-five bacterial isolates were isolated from untreated water samples collected from four water stations (El-Filal Station, El-Haras station, Bata station and Qalyub Station). Distinct colonies obtained from these samples were subjected to preliminary identification based on morphological and biochemical characteristics Among the isolates, four bacterial strains demonstrated resistance to 100% of the tested antibiotics, qualifying them as multi-drug resistant (MDR) bacteria. Advanced identification using the VITEK 2 compact system classified these MDR isolates as Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and Enterobacter aerogenes.