In Italy, serological screening is recommended to prevent congenital toxoplasmosis as part of the antenatal care protocol. Our study investigates (1) adherence to screening among Italian and migrant women and (2) specific T. gondii seroprevalence among hospitalized puerperas in Brescia and Udine, in northern Italy. All migrants (Group B) and a random Italian sample (Group A) filled in a questionnaire. Serological screening was rated as adequate when performed before conception or by the 12th week of gestation, and periodically repeated during pregnancy whenever negative. Nine hundred and twenty-two (922) puerperas were enrolled (Group A: 743; Group B: 179). Mean gestational age at first antenatal visit was 9.3 week, significantly more delayed in migrants (11.2w vs 8.9w; P < 0.0001). Toxoplasmosis was mentioned as a potential vertically transmitted infection by 380/922 (41.2%), but only by 13.4% of migrants (P < 0.0001). Anti-Toxoplasma IgG-Ab tested positive in 319/892 (35.8%), while the information was missing for 9 and 21 women resulted untested. Patients from northern Africa had an higher (AOR 3.63%; P = 0.002), while Asian patients a lower (AOR 0.33; P = 0.045) probability of being immune. A late screening was recorded in 115/848 (13.6%) women (Group A: 9.35%; Group B: 31.9%; P < 0.0001) and 82.1% of eligible migrants were not correctly monitored for toxoplasmosis during pregnancy. A late toxoplasma serological test in migrant women precludes the timely application of preventive measure and may represent an indicator of suboptimal antenatal care.