OBJECTIVE The cosmopolite protozoan, Toxoplasma gondii, has a significant economic and medical impact. Cats traditionally play a predominant role in the disease maintenance cycle; however, humans can be infected as a result of milk and meat consumption of Toxoplasma-infected livestock. In addition, infected pregnant women, even symptomless, can pass the disease to their unborn fetus. The limited clinical records and absence of specific national educational programs in countries like Egypt underscore the need for periodic toxoplasmosis disease evaluation. Here, we identified T. gondii seroprevalence among sheep and pregnant women and the associated risk factors in El-Minya Governorate, Egypt. METHODS Using peripheral blood, we detected T. gondii-specific antibodies in 151 sheep and 96 pregnant women sera from El-Minya Governorate using latex agglutination and indirect enzyme-linked immunosorbent assay. The impact of different environmental and behavioral risk factors identified with in-person interviews and serology results on acquiring toxoplasmosis was statistically analyzed. RESULTS The overall toxoplasmosis seroprevalence was 39.1% and 22.9% in sheep and pregnant women, respectively. Significantly higher seroprevalence was correlated with increasing sheep age and geographical location. Nonetheless, no statistical significance was found based on abortion history and pregnancy status of the examined sheep. Exposure factors important for pregnant women included pregnancy trimester, contact with cats, and the habit of eating undercooked sheep meat, which all had a statistically significant association with Toxoplasma seropositivity. CONCLUSIONS The current study confirms increased antibodies against toxoplasmosis in both sheep and pregnant women in El-Minya Governorate and a clear association between women's age, contact with cats, and the habit of eating undercooked sheep meat and seroreactivity to T. gondii. These results strongly suggest the need for a more comprehensive epidemiological study and public health awareness education for toxoplasmosis.