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Background  Certain salivary proteins of phlebotomine sand flies injected into the host skin during blood-feeding are highly antigenic and elicit strong antibody-mediated immune responses in repeatedly-exposed hosts. These antibodies can be measured by enzyme-linked immuno sorbent assays (ELISAs) using salivary gland homogenates (SGHs) as the source of antigens and serve as a markers for exposure to biting sand flies. Large-scale screening for anti-sand fly saliva antibodies requires replacement of SGH with recombinant salivary proteins. In East Africa,  Phlebotomus orientalis  is the main vector of  Leishmania donovani , a trypanosomatid parasite causing visceral leishmaniasis. We tested recombinant salivary proteins derived from  Ph .  orientalis  saliva to study exposure of domestic animals to this sand fly species.    Methodology/Principal Findings  Antigenic salivary proteins from  Ph .  orientalis  were identified by immunoblot and mass spectrometry. Recombinant apyrase rPorSP15, yellow-related protein rPorSP24, ParSP25-like protein rPorSP65, D7-related protein rPorSP67, and antigen 5-related protein rPorSP76 were tested using ELISA with sera of domestic animals from  L .  donovani  foci in Ethiopia where  Ph .  orientalis  is present. Our results highlighted recombinant yellow-related protein rPorSP24 as the most promising antigen, displaying a high positive correlation coefficient as well as good sensitivity and specificity when compared to SGH. This recombinant protein was the most suitable one for testing sera of dogs, sheep, and goats. In addition, a different antigen, rPorSP65 was found efficacious for testing canine sera.    Conclusions/Significance  Recombinant salivary proteins of  Ph .  orientalis , specifically rPorSP24, were shown to successfully substitute SGH in serological experiments to measure exposure of domestic animals to  Ph .  orientalis , the vector of  L .  donovani . The results suggest that rPorSP24 might be a suitable antigen for detecting anti- Ph .  orientalis  antibody-mediated reactions also in other host species.

Recombinant Salivary Proteins of Phlebotomus orientalis are Suitable Antigens to Measure Exposure of Domestic Animals to Sand Fly Bites