One of the most prominent concerns for the water consumers is microbial contamination, which is linked to various sources and health related problems, mainly to immunocompromised individual. Therefore the present study was focused on screening of drinking water mycobiota. One hundred and eighty samples were collected from 30 sites in Sohag Governorate, Egypt. Temperature, pH, total dissolved solids (TDS) and electric conductivity (EC) were investigated for each tested sample. The population density of yeast and filamentous fungi were detected and identified using YPDA And PDA media at 28±2° C. Candida glabrata was the most common yeast species in surface water represented 60% and 88% in winter and 51% and 100% in summer. Whereas in groundwater occurred in 43% and 32% in winter and 31% and 41% in summer of total yeast count and total tested samples, respectively. On the other hand, Aspergilllus species were the most widespread filamentous fungi in tested water samples. It was isolated from surface water samples in 84% and 100% in winter and 79% and 100% in summer, while in groundwater they were recorded in 95% and 100% in winter and 93% and 100% in summer of total fungal count and total tested samples, respectively. A. flavus was represented by 43% and 100% in winter and 38% and 100% in summer from surface water and 47% and 82% in winter and 43% and 82% in summer from groundwater. A. niger ranked the second place with 16% and 63% in winter and 15% and 75% in summer from surface water and 30% and 91% in winter and 28% and 91% in summer from groundwater.