title: Detection of Leishmania infantum in animals and their ectoparasites by conventional PCR and real time PCR.
abstract: Visceral leishmaniosis (VL) is a parasitic disease caused by Leishmania infantum, which is primarily transmitted by phlebotomine sandflies. However, there has been much speculation on the role of other arthropods in the transmission of VL. Thus, the aim of this study was to assess the presence of L. infantum in cats, dogs and their ectoparasites in a VL-endemic area in northeastern Brazil. DNA was extracted from blood samples and ectoparasites, tested by conventional PCR (cPCR) and quantitative real time PCR (qPCR) targeting the L. infantum kinetoplast DNA. A total of 280 blood samples (from five cats and 275 dogs) and 117 ectoparasites from dogs were collected. Animals were apparently healthy and not previously tested by serological or molecular diagnostic methods. Overall, 213 (76.1 %) animals and 51 (43.6 %) ectoparasites were positive to L. infantum, with mean parasite loads of 795.2, 31.9 and 9.1 fg in dogs, cats and ectoparasites, respectively. Concerning the positivity between dogs and their ectoparasites, 32 (15.3 %) positive dogs were parasitized by positive ectoparasites. The overall concordance between the PCR protocols used was 59.2 %, with qPCR being more efficient than cPCR; 34.1 % of all positive samples were exclusively positive by qPCR. The high number of positive animals and ectoparasites also indicates that they could serve as sentinels or indicators of the circulation of L. infantum in risk areas.
authors.affiliation: Department of Immunology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation, Av. Prof. Moraes Rego s/n, Recife, PE 50670-420, Brazil.
authors.forename: Rayana Carla Silva
authors.initials: RC
authors.lastname: de Morais
authors.forename: Suênia da Cunha
authors.initials: Sda C
authors.lastname: Gonçalves
authors.forename: Pietra Lemos
authors.initials: PL
authors.lastname: Costa
authors.forename: Kamila Gaudêncio
authors.initials: KG
authors.lastname: da Silva
authors.forename: Fernando José
authors.initials: FJ
authors.lastname: da Silva
authors.forename: Rômulo Pessoa E
authors.initials: RP
authors.lastname: Silva
authors.forename: Maria Edileuza Felinto
authors.initials: ME
authors.lastname: de Brito
authors.forename: Sinval Pinto
authors.initials: SP
authors.lastname: Brandão-Filho
authors.forename: Filipe
authors.initials: F
authors.lastname: Dantas-Torres
authors.forename: Milena
authors.initials: M
authors.lastname: de Paiva-Cavalcanti
mesh_terms: D000818:Animals; D001938:Brazil; D002415:Cats; D058251:Ctenocephalides / Q000469:parasitology*; D004285:Dogs; D018314:Leishmania infantum / Q000302:isolation & purification*; D007898:Leishmaniasis, Visceral / Q000635:transmission*; D008003:Phthiraptera / Q000469:parasitology*; D060888:Real-Time Polymerase Chain Reaction; D048508:Rhipicephalus sanguineus / Q000469:parasitology*