Phlebotomus perniciosus is a major vector of Leishmania infantum in the Mediterranean. While the seroprevalence of leishmaniosis in Spanish dogs and cats has been studied, data on the exposure of cats to P. perniciosus bites under natural conditions without repellents is limited. Stray cats could serve as sentinels for L. infantum and P. perniciosus exposure. This study analyzed sera from 204 apparently healthy stray cats, collected from January 2021 to January 2022, for antibodies against P. perniciosus saliva and L. infantum parasites. Anti-sand fly antibodies were detected in 40.69% of cats using an ELISA with the recombinant salivary protein SP03B of P. perniciosus. Seroprevalence of L. infantum infection was 23.52% by Western blot and 27.41% by ELISA, with an overall seroprevalence of 40.69% (95% CI 34.18-47.54%). This is the first assessment of antibody response to P. perniciosus saliva and L. infantum in naturally exposed stray cats in Spain. Further research is needed to examine the salivary antigens recognized by cats and to explore the relationship between P. perniciosus exposure and L. infantum infection severity in cats.

• Keywords
• Cat, ELISA, Leishmania infantum, Phlebotomus perniciosus, serology, western blotting,
• MeSH
• Enzyme-Linked Immunosorbent Assay veterinary   MeSH
• Insect Vectors parasitology   MeSH
• Cats MeSH
• Leishmania infantum * immunology   MeSH
• Leishmaniasis, Visceral * veterinary   epidemiology   immunology   MeSH
• Cat Diseases * epidemiology   parasitology   immunology   MeSH
• Phlebotomus * parasitology   immunology   MeSH
• Antibodies, Protozoan * blood   MeSH
• Seroepidemiologic Studies MeSH
• Animals MeSH
• Check Tag
• Cats MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Spain epidemiology   MeSH
• Names of Substances
• Antibodies, Protozoan * MeSH

Equids may be infected by zoonotic Leishmania spp., including Leishmania infantum, in regions where canine leishmaniasis (CanL) is endemic, and Leishmania martiniquensis, which has been reported in horses from Central Europe. This study was designed to evaluate the occurrence of both Leishmania spp. among equids living in CanL endemic areas of Italy, as well as to identify dipteran vectors from the same habitats. From March to October 2023, blood, serum and tissue samples from skin lesions were collected from equids (n = 98; n = 56 donkeys and n = 42 horses) living in Italy, as well as sand flies and biting midges. Blood samples (n = 98) and skin lesions (n = 56) were tested for Leishmania spp. by conventional and real time PCRs and sera were tested by immunofluorescence antibody tests (IFAT) for both L. infantum and L. martiniquensis. Insects were morphologically identified, and female specimens (n = 268 sand flies, n = 7 biting midges) analyzed for Leishmania DNA, as well as engorged sand flies (n = 16) for blood-meal detection. Two animals with skin lesions (i.e., one donkey and one horse) scored positive for Leishmania spp. DNA, and 19 animals (i.e., 19.4%; n = 13 donkeys and n = 6 horses) were seropositive for L. infantum, with five of them also for L. martiniquensis. Most seropositive animals had no dermatological lesions (i.e., 68.4%) while both animals molecularly positive for Leishmania spp. scored seronegative. Of the 356 sand flies collected, 12 females (i.e., n = 8 Sergentomyia minuta; n = 3 Phlebotomus perniciosus, n = 1 Phlebotomus perfiliewi) were positive for Leishmania spp. DNA, and one out of seven biting midges collected was DNA-positive for L. infantum. Moreover, engorged sand flies scored positive for human and equine DNA. Data suggest that equids living in CanL endemic areas are exposed to Leishmania spp., but their role in the circulation of the parasite needs further investigations.

• MeSH
• Ceratopogonidae parasitology   MeSH
• Endemic Diseases veterinary   MeSH
• Equidae * parasitology   MeSH
• Insect Vectors * parasitology   MeSH
• Horses parasitology   MeSH
• Leishmania infantum isolation & purification   genetics   MeSH
• Leishmania * isolation & purification   genetics   classification   MeSH
• Leishmaniasis * veterinary   epidemiology   parasitology   transmission   MeSH
• Horse Diseases parasitology   epidemiology   MeSH
• Dog Diseases * parasitology   epidemiology   transmission   MeSH
• Dogs MeSH
• Psychodidae parasitology   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Dogs MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Italy epidemiology   MeSH

Antibodies against Phlebotomus perniciosus sandfly salivary gland homogenate (SGH) and recombinant protein rSP03B, sandfly-borne Toscana virus (TOSV), Sandfly Fever Sicilian virus (SFSV) and Leishmania, as well as DNA of the latter parasite, were investigated in 670 blood samples from 575 human donors in Murcia Region, southeast Spain, in 2017 and 2018. The estimated SGH and rSP03B seroprevalences were 69% and 88%, respectively, although correlation between test results was relatively low (ρ = 0.39). Similarly, TOSV, SFSV and Leishmania seroprevalences were 26%, 0% and 1%, respectively, and Leishmania PCR prevalence was 2%. Prevalences were significantly greater in 2017, overdispersed and not spatially related to each other although both were positively associated with SGH but not to rSP03B antibody optical densities, questioning the value of the latter as a diagnostic marker for these infections in humans.

• Keywords
• Leishmania infantum, anti-saliva antibodies, blood donors, sandflies, sandfly fever sicilian virus, toscana virus,
• MeSH
• Blood Donors MeSH
• Leishmania infantum * MeSH
• Leishmaniasis * parasitology   veterinary   MeSH
• Humans MeSH
• Phlebotomus * parasitology   MeSH
• Antibodies MeSH
• Psychodidae * MeSH
• Recombinant Proteins MeSH
• Sandfly fever Naples virus * genetics   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Spain epidemiology   MeSH
• Names of Substances
• Antibodies MeSH
• Recombinant Proteins MeSH

BACKGROUND: In endemic areas of zoonotic leishmaniosis caused by L. infantum, early detection of Leishmania infection in dogs is essential to control the dissemination of the parasite to humans. The aim of this study was to evaluate the serological and/or molecular diagnostic performance of minimally and non-invasive samples (conjunctiva cells (CS) and peripheral blood (PB)) for monitoring Leishmania infection/exposure to Phlebotomus perniciosus salivary antigens in dogs at the beginning and the end of sand fly seasonal activity (May and October, respectively) and to assess associated risks factors. METHODS: A total of 208 sheltered dogs from endemic areas of leishmaniosis were screened. Leishmania DNA detection in PB on filter paper and CS was performed by nested-PCR (nPCR), while the detection of anti-Leishmania antibodies was performed using IFAT and ELISA. The exposure to P. perniciosus salivary antigens (SGH, rSP01 and rSP03B + rSP01) was measured by ELISA. RESULTS: Ninety-seven (46.6%) and 116 (55.8%) of the 208 dogs were positive to Leishmania antibodies or DNA by at least one test at the beginning and end of the sand fly season, respectively. IFAT and ELISA presented a substantial agreement in the serodiagnosis of leishmaniosis. Discrepant PB nPCR results were obtained between sampling points. Leishmania DNA was detected in CS of 72 dogs at the end of the phlebotomine season. The presence of antibodies to the parasite measured by ELISA was significantly higher in dogs presenting clinical signs compatible with leishmaniosis at both sampling points. Phlebotomus perniciosus salivary antibodies were detected in 179 (86.1%) and 198 (95.2%) of the screened dogs at the beginning and end of the phlebotomine season, respectively. CONCLUSIONS: The association between ELISA positivity and clinical signs suggests its usefulness to confirm a clinical suspicion. CS nPCR seems to be an effective and non-invasive method for assessing early exposure to the parasite. PB nPCR should not be used as the sole diagnostic tool to monitor Leishmania infection. The correlation between the levels of antibodies to P. perniciosus saliva and Leishmania antibodies suggests the use of a humoral response to sand fly salivary antigens as biomarkers of L. infantum infection.

• Keywords
• Blood, Conjunctival cells, Dog, Exposure, L. infantum, Phlebotomus pernicious, Saliva,
• MeSH
• Antigens, Protozoan immunology   MeSH
• Endemic Diseases prevention & control   MeSH
• Insect Vectors parasitology   MeSH
• Insect Proteins immunology   MeSH
• Immunoglobulin G blood   MeSH
• Conjunctiva cytology   parasitology   MeSH
• Insect Bites and Stings MeSH
• Leishmania infantum isolation & purification   MeSH
• Leishmaniasis blood   immunology   veterinary   MeSH
• Dog Diseases parasitology   prevention & control   transmission   MeSH
• Phlebotomus parasitology   MeSH
• Antibodies, Protozoan blood   MeSH
• Protozoan Proteins immunology   MeSH
• Dogs MeSH
• Risk Factors MeSH
• Serologic Tests MeSH
• Salivary Proteins and Peptides immunology   MeSH
• Animals MeSH
• Check Tag
• Dogs MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Antigens, Protozoan MeSH
• Insect Proteins MeSH
• Immunoglobulin G MeSH
• Antibodies, Protozoan MeSH
• Protozoan Proteins MeSH
• Salivary Proteins and Peptides MeSH

Phlebotomine sand fly-borne pathogens such as Leishmania spp. and phleboviruses are emerging threats to humans and animals worldwide. The aim of this work was to evaluate the exposure of cats from Portugal to Toscana virus (TOSV) and Sandfly Fever Sicilian virus (SFSV) and assess the associated risk factors. The possible association between exposure to Phlebotomus perniciosus saliva with TOSV and SFSV was also investigated. Out of 369 cats tested, 18 (4.9%, n = 365) were seropositive for TOSV, and eight (2.2%, n = 367) were seropositive for SFSV. Multivariate logistic regression analysis showed that cats presenting clinical signs that were compatible with leishmaniosis and antibodies to TOSV had a significantly higher risk of being SFSV seropositive. The presence of antibodies to sand fly-borne viruses in cats indicate that these animals are frequently exposed to sand flies and transmitted pathogens. Data suggest that cats can be used to qualitatively monitor human exposure to TOSV and SFSV in endemic areas. The clinical impact of SFSV in cats' health should be investigated. The identification of the sand fly species responsible for the circulation of TOSV and SFSV in nature and the evaluation of the vectorial competence of P. perniciosus to SFSV should also be addressed.

• Keywords
• Bunyavirales, Phenuiviridae, Phlebotomus perniciosus, Phlebovirus, Sandfly Fever Sicilian Virus, Toscana virus, arbovirus, cat, saliva,
• Publication type
• Journal Article MeSH