Abstract: Background and Objective: Toxoplasma gondii is widely spread protozoan parasite transmitted to human through various routes including blood transfusion. There is scarce information about the epidemiology of T. gondii in blood donors in Egypt. Therefore, this study aimed to determine the prevalence of T. gondii and associated sociodemographic and behavioral risk factors in a population of healthy blood donors of Beni-Suef Governorate and detect the possible risk for transmitting toxoplasmosis to blood recipients in a cross-sectional study. Materials and Methods: Blood samples were collected within 7 months from a total of 276 blood donors after taking questionnaire sheet to explore different risk factors. All participants were tested for presence of T. gondii IgG and IgM immunoglobulins using ELISA. Also seropositives were tested for IgG avidity test for confirming acute infection. Results: Out of 276 participants, 150 (54.3%) and 17 (6.1%) tested positive for anti-Toxoplasma IgG and IgM, respectively. Low avidity antibodies were detected in 6 (4%) out of 150 seropositive samples by IgG avidity test suggesting recent infection. Toxoplasma seropositivity was statistically significant in male patients, those living in rural areas and those eating improperly cooked/processed meat. However, there were no significant differences with age, educational level, blood group type or Rh factor, eating raw vegetables, contact with soil or cats. Conclusion: Toxoplasma seropositivity in blood donors may be an indication for its general prevalence in Beni-Suef Governorate. Presence of low avidity antibodies drew attention to the possible transmission of toxoplasmosis through blood transfusion to seronegative recipients and highlights the importance of Toxoplasma screening as a preceding test before donation of blood.