Toxoplasmosis has been associated with economic and public health concerns due to abortion in humans and animals. Detection for the presence of anti-Toxoplasma gondii antibodies in sera of 174 healthy sheep (IgG) and sera of 89 aborted women (IgG, IgM, and IgG avidity) using enzyme-linked immunosorbent assay and T. gondii in 59 sheep milk samples identified by PCR. The primary risk factors for sheep and women seropositive samples were also analyzed. Relatively higher seroprevalence of T. gondii, 62.6%, was recorded in sheep than 57.3% in aborted women. The IgM/IgG antibody responses in aborted women were 42.7%, 41.6%, 15.7%, and 0% for IgM -ve/IgG -ve (negative toxoplasmosis), IgM -ve/IgG +ve (chronic toxoplasmosis), IgM +ve/IgG +ve (maternal infection) and IgM +ve/IgG -ve (acute infection), respectively. IgM and IgG-positive samples showed a low avidity rate 14.3%, denoting acute maternal infection, while a much higher rate 78.6%, denoting chronic maternal infection. Molecular investigation of T. gondii in sheep milk samples revealed that 28.8% were positive by PCR at 470-bp. The respectable significant association between many risk factors and the seroprevalence of T. gondii in humans and sheep was recorded. The current study indicates elevated toxoplasmosis antibodies in sheep and aborted women from El-Beheira and Alexandria governorates, Egypt Additionally, it demonstrates a correlation between age, mutton intake, miscarriage, cat exposure, drinking water source, and breeding practices with seropositivity to T. gondii. These results largely confirm the need for greater in-depth toxoplasmosis epidemiological research and public health education initiatives.