The frequency of sandfly-host contacts can be measured by host antibody levels against sandfly salivary proteins. Recombinant salivary proteins are suggested to represent a valid replacement for salivary gland homogenate (SGH); however, it is necessary to prove that such antigens are recognized by antibodies against various populations of the same species. Phlebotomus perniciosus (Diptera: Psychodidae) is the main vector of Leishmania infantum (Trypanosomatida: Trypanosomatidae) in southwest Europe and is widespread from Portugal to Italy. In this study, sera were sampled from naturally exposed dogs from distant regions, including Campania (southern Italy), Umbria (central Italy) and the metropolitan Lisbon region (Portugal), where P. perniciosus is the unique or principal vector species. Sera were screened for anti-P. perniciosus antibodies using SGH and 43-kDa yellow-related recombinant protein (rSP03B). A robust correlation between antibodies recognizing SGH and rSP03B was detected in all regions, suggesting substantial antigenic cross-reactivity among different P. perniciosus populations. No significant differences in this relationship were detected between regions. Moreover, rSP03B and the native yellow-related protein were shown to share similar antigenic epitopes, as canine immunoglobulin G (IgG) binding to the native protein was inhibited by pre-incubation with the recombinant form. These findings suggest that rSP03B should be regarded as a universal marker of sandfly exposure throughout the geographical distribution of P. perniciosus.

Department of Biomedical and Medical Sciences University of Algarve Faro Portugal

Department of Parasitology Faculty of Science Charles University Prague Prague Czech Republic

Department of Veterinary Medicine and Animal Production University Federico 2 Naples Italy

Department of Veterinary Medicine University of Perugia Perugia Italy

Faculty of Veterinary Medicine Lusophone University of Humanities and Technologies Lisbon Portugal

Global Health and Tropical Medicine Medical Parasitology Unit Instituto de Higiene e Medicina Tropical New University of Lisbon Lisbon Portugal

Unit of Vector Borne Diseases and International Health Istituto Superiore di Sanità Rome Italy

Warwick Infectious Disease Epidemiology Research School of Life Sciences University of Warwick Coventry U K

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