Toxoplasma gondii is an important protozoan with serious obstetric impact in animals and women. The work aimed to relate public health significance of toxoplasmosis among some animals and women in Menoufia Governorate. A total 50 blood samples were collected from 50 adult sheep (30 from Kafr Tanbidi and 20 from Shebein El-Kom) and a total of 31 cattle milk samples were collected from different localities in Shebein El-Kom. Sheep sera were examined by ELISA IgG and recorded seropositivity of 60% (30/50) with 66.67% (20/30) were positive in Kafr Tanbidi and 50% (10/20) in Shebein El-Kom. PCR on sheep blood recorded 22% (11/50), of them 20% (6/30) were positive in Kafr Tanbidi and 25% (5/20) in Shebein El-Kom while PCR on cattle milk revealed 19.35% (6/31) positive for Toxoplasma DNA. On the other hand, Toxoplasma seroprevalence among 250 women was 54.4% (136/250) by ELISA IgG and 8.4% (21/250) by ELISA IgM. There was a significant association between IgG seroprevalence and age, increased gravidity, abortion, climate, cat contact, consumption of raw cow milk, undercooked meat and contaminated water from wells and between IgM seroprevalence and age, abortion and climatic conditions prevailed during sampling.