This study investigated the seroprevalence of Toxoplasma gondii in cattle in Gyeongbuk province, the largest producer of cattle in Korea. Of blood samples obtained from 568 animals, 0.5% (3 of 568) tested seropositive for T. gondii by using a commercial ELISA kit. Among young individuals (≤1 year old), adult individuals (2 years old), and old individuals (≥3 years old), 0 (0%) of 37, 3 (0.6%) of 474, and 0 (0%) of 57 were seropositive, respectively. Among male, female, and castrated individuals, 0 (0%) of 22, 0 (0%) of 74, and 3 (0.6%) of 472 were seropositive, respectively. Among individuals from east, south, and northwest regions of Gyeongbuk province, 0 (0%) of 155, 2 (0.7%) of 288, and 1 (0.8%) of 125 were seropositive, respectively. No statistical differences were observed among the groups. Although a low T. gondii seroprevalence was detected in cattle raised in Gyeongbuk province, toxoplasmosis is still a concern owing to the Korean habit of eating raw or undercooked meat and the consequent risk of Toxoplasma transmission to animals and humans.