Background: Knowledge of toxoplasmosis has been assessed among obstetricians, medical students, midwifes and pregnant women. The aims of the study were as follows: 1) evaluation of intra- and inter-group variation in correct answers to the questions included in the questionnaire used as a study tool; 2) evaluation of inter-group variation in answers to specialist and non-specialist questions; and 3) intra-group evaluation of self-assessed difficulty in completing the questionnaire. Materials and Methods: In the prospective study, 310 participants were included: 109 pregnant women, 116 midwifes 85 physicians (including 69 specialists in OB/GYN) and 16 medical students. The anonymous questionnaire consisted of 26 questions, with four answers proposed for each question of which one was correct. Results: There was no difference between correct answers among midwifes, physicians and medical students (p = 0.20), but the difference was found between the group of physicians and medical students and pregnant women as well as between midwifes and pregnant women (p < 0.001). Physicians, medical students and midwifes gave best answers to the non-specialist questions (p < 0.001), and midwifes obtained higher scores than pregnant women (p < 0.001). The mean percentage of correct answers to the specialist questions was among pregnant women (29%) showing a high level of randomness. The best answers to the non-specialist questions were found for physicians and midwifes (p = 0.93). However, there was a difference between the group of physicians and medical students and pregnant women and between midwifes and pregnant women in self-assessment of difficulty in completing the questionnaire (p < 0.001). Conclusions: Pregnant women assessed their knowledge of toxoplasmosis as least adequate, whereas midwifes. as most adequate. Pregnant women's knowledge about toxoplasmosis is poor. The group of physicians and medical students yielded evidently better score than pregnant women. There is a need for the improvement among prenatal care providers in both patient education and self-education. Med Pr 2010;61(3):271-276