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Background  Phlebotomine sand flies are blood-sucking insects that can transmit  Leishmania  parasites. Hosts bitten by sand flies develop an immune response against sand fly salivary antigens. Specific anti-saliva IgG indicate the exposure to the vector and may also help to estimate the risk of  Leishmania  spp. transmission. In this study, we examined the canine antibody response against the saliva of  Phlebotomus perniciosus , the main vector of  Leishmania infantum  in the Mediterranean Basin, and characterized salivary antigens of this sand fly species.    Methodology/Principal Findings  Sera of dogs bitten by  P. perniciosus  under experimental conditions and dogs naturally exposed to sand flies in a  L. infantum  focus were tested by ELISA for the presence of anti- P. perniciosus  antibodies. Antibody levels positively correlated with the number of blood-fed  P. perniciosus  females. In naturally exposed dogs the increase of specific IgG, IgG1 and IgG2 was observed during sand fly season. Importantly,  Leishmania -positive dogs revealed significantly lower anti- P. perniciosus  IgG2 compared to  Leishmania -negative ones. Major  P. perniciosus  antigens were identified by western blot and mass spectrometry as yellow proteins, apyrases and antigen 5-related proteins.    Conclusions  Results suggest that monitoring canine antibody response to sand fly saliva in endemic foci could estimate the risk of  L. infantum  transmission. It may also help to control canine leishmaniasis by evaluating the effectiveness of anti-vector campaigns. Data from the field study where dogs from the Italian focus of  L. infantum  were naturally exposed to  P. perniciosus  bites indicates that the levels of anti- P. perniciosus  saliva IgG2 negatively correlate with the risk of  Leishmania  transmission. Thus, specific IgG2 response is suggested as a risk marker of  L. infantum  transmission for dogs.

Canine Antibody Response to Phlebotomus perniciosus Bites Negatively Correlates with the Risk of Leishmania infantum Transmission