Neutralizing antibodies to sandfly fever Naples, sandfly fever Sicilian and Toscana viruses were investigated among 479 sera collected from a normal human population in Cyprus. Antibody prevalence rates of 57%, 32% and 20% were found to Naples, Sicilian and Toscana viruses, respectively. The observed frequency of dual and triple infections was higher than would be expected with a random chance of infection. Antibody prevalence rates were similar for men and women for all three viruses tested, but one of two study sites had significantly higher antibody prevalence to Naples and Sicilian viruses than the other. Individuals with antibodies to both Naples and Toscana viruses had higher antibody levels to Naples virus than those with antibodies to Naples virus only. If the antibody prevalence rates found in this study reflect a history of clinical disease as described in the literature, sandfly fever poses a significant public health problem in Cyprus.