Background: Toxoplasmosis is a major opportunistic infection caused by single-cell protozoan from the Coccidia family. Up to 95% of infected individuals remain asymptomatic, nevertheless, it may be a leading cause of foodborne illness-related hospitalisation and death. Primo-infection can cross the placental barrier and cause life-threatening sequelae for the fetus leading to miscarriage, hydrocephalus and, retinochoroiditis. Moreover, the infection may cause infertility, endometritis, and uterine atrophy and, impaired folliculogenesis for the mother. Aim: To evaluate the effectiveness of an educational program on perception and anxiety level regarding toxoplasmosis among seropositive pregnant women in Beni-Suef in Egypt. Subjects and methods: A quasi-experimental design was used with 50 seropositive pregnant women attending the inpatient/outpatient antenatal care units. Tools: A structured knowledge questionnaire, pre-test/post-test/retained-test knowledge assessment sheet and Zung's self-rating anxiety scale. Results: Pre-program 96% of women scored poor knowledge and no participants (0.00%) reported severe anxiety; whereas, immediately post-program 70% of women scored good knowledge and 84% reported severe anxiety. After implementation of the program, all women (100%) gained a good knowledge score while 48% of them reported severe anxiety. Significant association between mean knowledge scores and demographic variables (p < .0001) was found. Conclusions: Although the program is effective to enhance women's perception regarding Toxoplasmosis, it exaggerated the anxiety level for pregnant women. A negative association was found between women's perception and anxiety level, however, this relation was not significant. Recommendations: Implementation of health education programs aimed at primary prevention of toxoplasmosis is recommended for all women of childbearing age.