title: The quest for canine leishmaniasis in Romania: the presence of an autochthonous focus with subclinical infections in an area where disease occurred.
abstract: Canine leishmaniasis is a severe, potentially life-threatening, systemic vector-borne disease of dogs caused by protozoan parasites of the genus Leishmania. Romania has been traditionally regarded as a non-endemic country for leishmaniasis with sporadic human disease cases. However, the recent report of an autochthonous canine leishmaniasis case (the first in the last 80 years) suggested the presence of an infection focus in the area of Râmnicu Vâlcea. The present study describes a survey of canine leishmaniasis in this geographical area with comparison to a georeferenced dataset of sand fly distribution based on historical literature records.
  
  
  
  The study was carried out in Râmnicu Vâlcea and included samples (serum, blood and conjunctival swabs) collected from 80 dogs including client-owned dogs from two local practices and dogs from two public shelters. Serum anti-leishmanial antibodies were assessed by ELISA. All blood and conjunctival samples were assessed by real-time quantitative PCR, targeting the leishmanial kinetoplast minicircle DNA.
  
  
  
  Three dogs (3.7 %) were seropositive and another four (5.0 %) showed borderline results indicative of exposure or infection. TaqMan PCR was performed for all dogs, on both blood and conjunctival swabs. Seven dogs (8.7 %) were positive by conjunctival swab PCR and one dog (1.2 %) by blood PCR. None of the positive dogs presented clinical signs compatible with canine leishmaniasis.
  
  
  
  This is the first study evaluating canine leishmaniasis in a dog population in Romania by both highly sensitive PCR and serology. Although the prevalence was relatively low compared to other endemic regions, our results clearly demonstrate the presence of a canine leishmaniasis focus in Romania.
authors.affiliation: Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania. mirabela.dumitrache@usamvcluj.ro.
authors.forename: Mirabela Oana
authors.initials: MO
authors.lastname: Dumitrache
authors.affiliation: Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, Israel.
authors.forename: Yaarit
authors.initials: Y
authors.lastname: Nachum-Biala
authors.affiliation: Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, Israel.
authors.forename: Matan
authors.initials: M
authors.lastname: Gilad
authors.affiliation: Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania.
authors.forename: Viorica
authors.initials: V
authors.lastname: Mircean
authors.affiliation: Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania.
authors.forename: Cristina Daniela
authors.initials: CD
authors.lastname: Cazan
authors.affiliation: Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania.
authors.forename: Andrei Daniel
authors.initials: AD
authors.lastname: Mihalca
authors.affiliation: Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, Israel.
authors.forename: Gad
authors.initials: G
authors.lastname: Baneth
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D004283:Dog Diseases / Q000453:epidemiology / Q000469:parasitology*; D004285:Dogs; D007896:Leishmaniasis / Q000453:epidemiology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D015995:Prevalence; D012383:Romania / Q000453:epidemiology
chemical_list: D000913:Antibodies, Protozoan
keywords: Canine leishmaniasis; ELISA; Leishmania infantum; PCR; Romania