Introduction: Toxoplasmosis is a widespread foodborne zoonosis that is usually benign. In cases of primary infection during pregnancy, it can lead to fetal complications that are sometimes severe. The objectives of this study were to estimate trends in the seroprevalence of toxoplasmosis and its associated factors among pregnant women in France between 1995 and 2010, using data from the National Perinatal Surveys (ENP). Methods: The study focused on the ENP surveys conducted in 1995, 2003, and 2010. The seroprevalence of toxoplasmosis was estimated for each survey, and a multivariate analysis was performed to identify factors associated with toxoplasmosis. Results: The analyses included 42,916 women in late pregnancy for whom serological status regarding toxoplasmosis was recorded. The seroprevalence of toxoplasmosis among pregnant women decreased over time, from 54.3% in 1995 to 43.8% in 2003 and to 36.7% in 2010. It is significantly associated with age, nationality, and region of residence, with an east-west geographic gradient in seroprevalence. Discussion: The seroprevalence of toxoplasmosis among pregnant women has decreased significantly between 1995 and 2010. A reassessment of the current screening strategy for toxoplasmosis during pregnancy should be considered.