The purpose of our study was to assess whether Leishmania infantum parasitemia occurs in asymptomatic Leishmania-seropositive subjects. Samples from 500 blood donors were tested using an enzyme-linked immunosorbent assay (ELISA). Anti-Leishmania antibodies were not found in any sample. Our findings suggest that the risk of L. infantum transmission by blood transfusion in Sicily is very low.