The present study is destined to measure the nitrate levels of groundwater in Sohag Governorate aiming to determine their possible sources and evaluate their environmental and health impact. 17% of the analyzed samples have NO3-levels go over the MAL. Potentially harmful levels of NO3-exceeding nearly four times higher than the maximum allowable limit (MAL) for drinking water (45 mg/l) were reported. About 32% of the examined samples displaying nitrate levels more than 13 mg/l (pollution indicator). Nitrate is converted to nitrite in the infant’s stomach causing transformation of hemoglobin (the oxygen carrier) to methemoglobin that can’t carry oxygen. Therefore, oxygen starvation might occur causing serious disorders or even death (methemoglobinemia). Moreover, nitrate is a precursor in the formation of the carcinogenic N-nitroso compounds (NOC). Also, the augmentation of nitrate in groundwater is of vital concern because they indicate the groundwater pollution by anthropogenic contaminants (chemical and/or biological). Quantification of the distribution pattern of the environmental diseases (eg methemoglobinemia, cancer, kidney failure, neurological disorders) in Sohag Governorate and its interrelation with the drinking water pollution is strongly recommended; the convenient regulations can be then taken.