Background Toxoplasmosis has public health importance, particularly in immunocompromised individuals such as pregnant women leading to congenital abnormalities and fetal losses. In this study, we aimed to determine the sero-prevalence of toxoplasmosis in sheep at Mandeeq Slaughter House in Hargiesa and pregnant women attending antenatal care at Hargiesa Group Hospital. Methods Cross-sectional study design with systematic random sampling method and Questionnaire surveys were used for the collection data from sheep and pregnant women. Sera were collected and examined for anti- Toxoplasma gondii antibodies using latex agglutination test. Results The overall prevalences of toxoplasmosis were 25.5% and 28% in sheep and human, respectively. Multivariable logistic regression analysis indicated that female (AOR = 2.18; 95% CI: 1.38 - 3.47; p = 0.001) and young age groups of sheep (AOR = 3.04; 95% CI: 1.04-8.86; p = 0.041) were significantly associated with Toxoplasma gondii sero-positivity. In pregnant women, age groups between 25-34 (AOR = 2. 76; 95% CI: 1.07 - 7.14; P = 0.037), pregnant women who have cats in their home (AOR = 6.45; 95% CI: 2.37-17.52; P = 0.000), women who have close contact with garden soil (AOR = 6.74; 95% CI: 2.55-17.81; P = 0.000), poor hand washing practices before food eating (AOR = 29.5; 95% CI: 5.41– 161.11; P= 0.000) and drinking tap water (AOR = 8.4; 95% CI: 2.54 – 28.08; P = 0.000) were significantly associated with Toxoplasma gondii sero-positivity. Conclusion This is the first report regarding to toxoplasmosis in Hargeisa, where pregnant women should avoid eating uncooked mutton and close contact with cats besides good hygienic practices related to food handling.