Thyroid volume (ThV) and echogenicity by ultrasound were estimated in 324 schoolchildren (aged between 10 and 13-years) from high nitrate area (HNA) located in agricultural lowland with high nitrate drinking water supply (51–274 mg/l) from shallow wells. The data were compared to children of the same age from low nitrate area (LNA) consisting of 168 children from the neighboring area with very low nitrate (<2 mg/l) drinking water and of 596 children from the city of Košice located in a vicinity of LNA and also supplied by low nitrate water. Blood samples were obtained from 315 willing children from HNA and 109 children from LNA and the levels of thyrotropin (TSH), total thyroxine (TT4), free triiodothyronine (FT3) and thyroperoxidase antibodies (anti-TPO) in serum were determined. ThV (mean ± SE) in 10-year (5.10 ± 0.14 ml) and 13-year (5.97 ± 0.11 ml) old children from HNA was significantly higher than that in two groups of respective age from LNA, 4.58 ± 0.17 ( p < 0.02) and 5.23 ± 0.15 ml ( p < 0.05), and from the city of Košice, 4.77 ± 0.10 ml ( p < 0.05) and 4.87 ± 0.10 ml ( p < 0.0001). The frequency of hypoechogenicity in HNA was also significantly higher than that in pooled LNA plus Košice, 13.7% vs. 4.7% ( p < 0.01) in 10-year and 10.6% vs. 5.7% ( p < 0.03) in 13-year, respectively. The frequency of TSH level in the range of subclinical hypothyroidism (>4.0 mU/l) in pooled age groups from HNA was 13/324 (4.0%) and that of positive anti-TPO was 8/324 (2.5%), while no case of either increased TSH or positive anti-TPO was found in 109 children from LNA. Finally, no differences in the levels of TT4 and FT3 were found between HNA and LNA. It was concluded that long-term exposure to high nitrate intake by drinking water and home made meals from local products results in increased thyroid volume and increased frequency of signs of subclinical thyroid disorders (thyroid hypoechogenicity by ultrasound, increased TSH level and positive anti-TPO).