Seroprevalence of Toscana virus in blood donors in mainland Portugal.

Background: Toscana virus (TOSV; Phlebovirus toscanaense), a phlebovirus transmitted by sand flies, is a growing public health concern in the Mediterranean region, with infections often being asymptomatic but potentially leading to neuroinvasive disease. Despite its presence in neighboring countries, data on TOSV seroprevalence in Portugal are limited. This study aimed to estimate the national seroprevalence of TOSV among blood donors in mainland Portugal and explore associations with sociodemographic factors and Leishmania infection.

Methods: A cross-sectional study was conducted using serum samples from 3593 blood donors across mainland Portugal, collected between February and June 2022. Anti-TOSV antibodies were detected via microneutralization assay, and anti-Leishmania antibodies had previously been tested using ELISA. Sociodemographic data were obtained from self-administered questionnaires. Seroprevalence was estimated by region, and multivariate logistic regression was used to identify factors associated with TOSV infection.

Results: Overall, the estimated national true seroprevalence of TOSV was 2.6% (95% CI 2.1-3.1%). Regional seroprevalence varied significantly, with the highest values (up to 14.8%) in Alto Alentejo, Baixo Alentejo, Douro, Alto Tâmega e Barroso and Oeste regions. Multivariate analysis showed that age ≥ 50 years (aOR 1.70, 95% CI 1.04-2.77), residing in the Alentejo region (aOR 3.05, 95% CI 1.85-5.02) and positive/borderline Leishmania serology (aOR 2.31, 95% CI 1.29-4.15) were significantly associated with TOSV infection.

Conclusions: This study highlights new areas of TOSV circulation in Portugal, particularly in regions with higher Leishmania seroprevalence and visceral leishmaniasis incidence, suggesting co-circulation of these pathogens. Although a lower seroprevalence was obtained compared to neighboring countries, TOSV should still be considered in the differential diagnosis of viral meningitis and encephalitis in Portugal, especially in potentially high-risk regions. Further research is needed to better understand the ecological drivers of TOSV distribution in Portugal.