This study was carried out to assess the fluoride concentration in groundwater in some villages of southeastern part of Anantapur district Andhra Pradesh, India, where groundwater is the main source of drinking water. The main objective of this study was to assess the association between water fluoride levels and prevalence of dental fluorosis among school children of Anantapur District, India. Fluoride in presently studied sites was recorded in the ranges of 0.5 and 5.7 mg/L. The fluoride concentration in the Mudigubba Mandal (1.7-5.7 mg/L) and in the Nallamada Mandal (0.5-4.8 mg/L), Kadiri Mandal (1.5-5.1 mg/L). The percentage of school children suffering from dental fluorosis are: 56.3% in Mudigubba; 38.4% in Nallamada and 52.3% in Kadiri Mandals. From this study, it has been found that Rall Ananathapuram, Sankepalli, Uppalapadu, Jonnalakothapalli, Mudigubba, Vankarakunta, Donnikota, Sanevaripalli, Vellamaddi, Nallamada, Pulagampalle, Patnam, Eguvapalli, Kadirikuntlapalli, Yerradoddi, Kutagulla and Alampur are the fluoride endemic villages, where the fluoride level in drinking water is above 1 mg/L. The results of the present study revealed that there exists a significant positive correlation between fluoride concentration in drinking water and dental fluorosis in high-fluoride villages. As per the desirable and maximum permissible limit for fluoride in drinking water, determined by WHO or by Bureau of Indian Standards, the groundwater of about 95% of the studied sites is not consumable for drinking purposes. Due to the higher fluoride level in drinking water several cases of dental fluorosis have appeared at alarming rate in this region. The result suggests that the groundwater should be used by the residents only after defluoridation