This study aimed to determine the prevalence and risk factors associated with Toxoplasma gondii infection in blood donors from the Brazilian Semiarid region, and to explore its implications for transfusion safety. Samples were collected from 646 donors at blood donation centers in the states of Ceará and Paraíba. Serological diagnosis was performed using BIOLISA TOXOPLASMOSE ELISA kits for anti-T. gondii IgM and IgG antibodies, and molecular diagnosis was conducted by conventional PCR targeting a 529-bp noncoding repetitive fragment. Epidemiological questionnaires on variables associated with infection were administered, and statistical analysis was performed in univariate and multivariate stages, using multiple logistic regression. Among the 646 donors, 43.4% (281/646) were positive for anti-T. gondii IgG antibodies, 0.3% (2/646) for IgM antibodies, and none tested positive by PCR. In the univariate analysis, age, family income, educational level, salad washing practices, water source, raw milk consumption, and duration as a donor were significantly associated, whereas in the multivariate analysis only “age” and “salad washing practices” remained significant. A substantial IgG seroprevalence was observed among blood donors in the Brazilian Semiarid. The low IgM frequency, concurrent IgG positivity, and negative PCR results are consistent with a low transfusion risk in the region. However, these findings should be interpreted cautiously, as negative PCR results do not completely rule out the presence of circulating parasites. Age was identified as a risk factor, whereas proper salad washing showed a protective effect.