Water quality assessment of River Nile has been studied in the Upper Egypt region between April 2011 and March 2012 to identify the relationship between the physicochemical parameters and microbiological characteristics. Thirty six water samples were collected during hot and cold seasons along the area extending from Tima to Dar-Elsalam cities, Sohag governorate, Egypt. Results indicated that the physicochemical parameters in all samples increased significantly in the hot season than cold season. In addition, the bacteriological assessment for water samples indicated that most of locations were polluted with faecal coliform and pathogenic bacteria which were identified as Escherichia coli, Salmonella spp., Pseudomonas aeruginosa and Shigella spp.