Seroprevalence of sand fly fever Sicilian virus in blood donors in mainland Portugal

Background: Sicilian virus (SFSV), a phlebovirus transmitted by sand flies, is an understudied arbovirus in the Mediterranean region, with limited data on its epidemiology and human health impact. This study aimed to estimate the seroprevalence of SFSV among blood donors in mainland Portugal and explore associations with sociodemographic factors and exposure to other sand-fly-borne pathogens. Methods: A cross-sectional study was conducted using serum samples from 800 blood donors collected between February and June 2022. The study sample was selected from a previously established cohort designed for Leishmania seroprevalence assessment. The microneutralization technique was employed to detect anti-SFSV antibodies. Sociodemographic data were obtained from self-administered questionnaires. Associations between SFSV seropositivity and Toscana virus (TOSV)/Leishmania seropositivity or sociodemographic variables were explored using univariate analysis and multivariate logistic regression. Results: Overall, the estimated national true seroprevalence of SFSV was 4.7% (95% CI 3.4–6.3%). Regional seroprevalence varied significantly, with the highest rates (up to 11.9%) observed in the Algarve, Alentejo, and Grande Lisboa regions, respectively. In univariate analysis, SFSV seropositivity was not significantly associated with sex, age, dog ownership, or positive serology for TOSV or Leishmania. In multivariate analysis, geographic area of residence was the only independent factor associated with seropositivity (adjusted odds ratio 3.05; 95% CI 1.85–5.02; p < 0.001). Discussion: TThis study represents the first nationwide SFSV seroprevalence estimate in Portugal, revealing wider circulation than previously recognized. The lack of association with TOSV or Leishmania seropositivity could suggest the involvement of distinct vector species. Conclusions: Given the observed geographic clustering, SFSV should be considered in the differential diagnosis of undifferentiated febrile syndromes, particularly in endemic regions during peak sand fly activity. Further research is needed to identify specific vectors, improve diagnostic capabilities, and assess the clinical impact of SFSV infections in Portugal.