Abstract Background The expectancy of Toxoplasma gondii transmitted from livestock and raw meat to humans is a public health problem and is an example of the One Health theory. Objectives This survey aimed to determine the seroprevalence and risk factors related to this common infection in individuals occupationally exposed (IOE) to livestock, raw meat and viscera in industrial slaughterhouses and livestock fields in Isfahan province, central Iran. Methods This study is a case–control survey carried out on the 401 serum samples of IOE (including slaughterhouse workers, butchers, veterinarians, veterinary technicians, livestock farmers and farm workers) compared to 401 archived samples of the general population (that all matched with cases by region, age and gender). All 802 samples were investigated for anti‐T. gondii IgM and anti‐T. gondii IgG using enzyme‐linked immunosorbent assay. Results A statistically significant higher anti‐T. gondii IgG occurrence (p