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Blood donors (n = 663) from the Nový Jičín district, Czech Republic, were examined for the presence of antibodies to Toxoplasma gondii. The indirect fluorescent antibody test was used to simultaneously detect IgM and IgG antibodies. Titres ≥ 20 were considered positive. The seroprevalence of IgM and IgG antibodies was 2.4% and 32.1%, respectively. Periods, for how long the blood donors were infected, are discussed.