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This study assessed the prevalence of co-infections (human immunodeficiency virus, hepatitis B, and syphilis) and associated risk factors for Toxoplasma gondii infection among pregnant women in Mampong Municipality, Ghana. A cross-sectional design was used to recruit 201 pregnant women from six health facilities conveniently. Participants' socio-demographics, clinical and environmental data were collected using a structured questionnaire. Using 2 ml of blood, T. gondii seroprevalence was determined by the TOXO IgG/IgM Rapid Test Cassette. Data was analyzed using descriptive and logistic regression analysis with SPSS version 27 to determine the prevalence and associations of T. gondii infection with other variables, respectively. The seroprevalence of T. gondii was 49.75%, of which 40.30%, 2.49%, and 6.97% tested positive for IgG, IgM, and IgG/IgM, respectively. Co-infection of toxoplasmosis with viral hepatitis B, human immunodeficiency virus (HIV), and syphilis rates were 15%, 1%, and 4%, respectively and were not risk factors for T. gondii transmission. Educational level and residential status were associated with toxoplasmosis [p < 0.05]. Participants with higher education had a reduced risk of T. gondii infections compared to a lower level of education [AOR = 0.39 (0.13, 0.99) p = 0.049]. Similarly, the risk of T. gondii infection was significantly lower among individuals residing in peri-urban [AOR = 0.13 (0.02-0.70), p = 0.02] and urban areas [AOR = 0.10 (0.02-0.78), p = 0.03] compared to those in rural areas. Backyard animals with extensive and semi-intensive systems, without veterinary care, and contact with animal droppings and water sources were significant risk factors for T. gondii infection [p < 0.05]. Miscarriage was associated with T. gondii infection [p < 0.05]. The burden of T. gondii infection was high among the study population, posing a risk of mother-to-child transmission. Key risk factors included low education, rural residence, backyard animal exposure, poor hygiene, and unsafe water sources. Toxoplasmosis was associated with miscarriage; thus, integrating it into routine antenatal screening could improve pregnancy outcomes. Health promotion interventions such as education on zoonotic risks, improved sanitation, safe water practices, and veterinary care for domestic animals are recommended to reduce infection risk among pregnant women.