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

Abstract

AbstractLeishmaniasis is a chronic global arthropod-borne zoonotic disease produced by several species ofLeishmania, with cutaneous, mucocutaneous, and visceral clinical manifestations. In Spain, onlyLeishmania infantumhas been reported so far, although other species ofLeishmania, such asL. tropicaandL. major, are present in surrounding countries.The aim of this work is to analyze the occurrence ofLeishmaniaspp. infection in European wildcats (Felis silvestris) as sentinels, including their genotypic characterization. Necropsies of 18 road killed wildcats were conducted. Samples of ear skin and spleen were taken for DNA isolation and PCR of the highly sensitiveSSUrDNAtarget. Subsequent PCR tests were performed using more specific targets for the determination ofLeishmaniaspecies:hsp70andITS1. Positive samples were sequenced, and phylogenetic trees constructed. Seven wildcats were found positive forLeishmaniaspp.. Based on thehsp70andITS1sequences, an animal was found to be infected only withL. tropicain ear skin samples, while two cats were found to be infected withL. infantumin both the ear skin and the spleen. In one animal, a clear sequence ofL. infantumITS1 and a sequence ofL. tropica hsp70were obtained from the ear skin. Since hsp70 and ITS1 sequencing was not possible in three cats, the species ofLeishmaniainfecting them was not determined.This is the first report of autochthonous infection withL. tropicain the Iberian Peninsula. Health care professionals, including physicians, dermatologists, and veterinarians, must be aware of this for a correct diagnosis, treatment, and management of possible co-infections.