Wildlife as a sentinel for pathogen introduction in nonendemic areas: first detection ofLeishmania tropicain wildlife in Spain

Abstract

Background: Leishmaniasis is a global arthropod-borne zoonotic disease of chronic course and cutaneous, mucocutaneous, and visceral clinical manifestations. In the Iberian Peninsula, only Leishmania infantum has been reported, although other species of Leishmania, such as L. tropica and L. major, are present in surrounding countries. Aim: The aim of this work is to analyse the occurrence of Leishmania spp. infection in wildcats as sentinels, including their genotypic characterization. Methods: Necropsies of 18 roadkill wildcats were carried out. Samples from ear skin and spleen were taken, and DNA isolation and PCR analysis of the highly sensitive SSUrDNA target were conducted. Subsequent PCR tests were conducted using more specific targets for Leishmania species determination: hsp70 and ITS1Positive samples were sequenced, and phylogenetic trees were constructed.Results: Seven out of 18 wildcats were found positive to Leishmania spp. in ear skin; of them, only two were positive in spleen samples. . Based on the hsp70 and ITS1 sequences, one animal was found to be infected solely with L. tropica in ear skin samples, while two cats were found to be infected with L. infantum in both ear and spleen. Another animal was found to be co-infected with L. infantum and L. tropica in ear skin. The Leishmania species could not be determined in three cats as the targeted SSUrDNA is only specific to the genus. Conclusions: This is the first report of autochthonous L. tropica infection in the Iberian Peninsula. Health care professionals, including physicians, dermatologist and veterinarians must be aware of this for a correct diagnosis, treatment, and management of possible co-infections.