AbstractBackgroundLeishmania infantumis a vector-borne pathogen endemic in countries in the Mediterranean basin, including Italy. Dogs act as the primary reservoir for this parasite, but other animal species may also be infected. Low-to-moderate seroprevalence levels of infection have been reported in apparent healthy equine populations in southern Europe, reinforcing the importance of exploring those species, including horses, that act as a food source for vectors and may thus participate in the epizoological scenario of canine leishmaniosis (CanL) and zoonotic visceral leishmaniosis (ZVL). Since little is known regarding the exposure toL. infantumin horses in Italy, we assessed the seroprevalence in healthy equine populations from different CanL endemic areas.MethodsThe survey was conducted on 660 apparently healthy horses distributed throughout central and northern regions of Italy between 2016 and 2019. Blood samples were collected and the presence of anti-Leishmaniaantibodies (IgG) was investigated by the immunofluorescence antibody test. Information on the location and altitude of the stables, along with the horses’ breed, age, sex, and reproductive status was obtained by filling in a questionnaire. This was then used for statistical analysis by generalized linear models to explore risk factors associated with seroreactivity toL. infantum.ResultsAn average seroprevalence of 13.9% was detected forL. infantumin the equine populations investigated, with statistically significant associations between seroprevalence, geographical variables (northernvscentral Italy, origin and altitude) and individual factors (i.e. age and breed morphotype).ConclusionsOur results highlight that horses are frequently exposed toL. infantum. Further prevalence surveys in horses, also using direct methods (e.g. PCR), are warranted to clarify the role of these hosts in the epidemiology ofLeishmaniain Italy.