title: Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs.
abstract: BACKGROUND
  The main transmission route of Leishmania infantum is through the bites of sand flies. However, alternative mechanisms are being investigated, such as through the bites of ticks, which could have epidemiological relevance. The objective of this work was to verify the presence of Leishmania spp. in Rhipicephalus sanguineus sensu lato collected from naturally infected dogs in the Federal District of Brazil.
  
  
  METHODS
  Ticks were dissected to remove their intestines and salivary glands for DNA extraction and the subsequent amplification of the conserved region of 120 bp of kDNA and 234 bp of the hsp70 gene of Leishmania spp. The amplified kDNA products were digested with endonucleases HaeIII and BstUI and were submitted to DNA sequencing. Isolated Leishmania parasites from these ticks were analyzed by multilocus enzyme electrophoresis, and the DNA obtained from this culture was subjected to microsatellite analyses.
  
  
  RESULTS
  Overall, 130 specimens of R. sanguineus were collected from 27 dogs. Leishmania spp. were successfully isolated in culture from five pools of salivary glands and the intestines of ticks collected from four dogs. The amplified kDNA products from the dog blood samples and from the tick cultures, when digested by HaeIII and BstUI, revealed the presence of L. braziliensis and L. infantum. One strain was cultivated and characterized as L. infantum by enzyme electrophoresis. The amplified kDNA products from the blood of one dog showed a sequence homology with L. braziliensis; however, the amplified kDNA from the ticks collected from this dog showed a sequence homology to L. infantum.
  
  
  CONCLUSIONS
  The results confirm that the specimens of R. sanguineus that feed on dogs naturally infected by L. infantum contain the parasite DNA in their intestines and salivary glands, and viable L. infantum can be successfully isolated from these ectoparasites.
authors.affiliation: Programa de Pós-Graduação em Medicina Tropical, Universidade de Brasília, Brasília, DF, Brazil. vivianemedeiros@gmail.com.
authors.forename: Viviane
authors.initials: V
authors.lastname: Medeiros-Silva
authors.affiliation: Programa de Pós-Graduação em Medicina Tropical, Universidade de Brasília, Brasília, DF, Brazil. rgurgel@unb.br.|Laboratório de Parasitologia Médica e Biologia de Vetores, Universidade de Brasília, Brasília, DF, Brazil. rgurgel@unb.br.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Gurgel-Gonçalves
authors.affiliation: Programa de Pós-Graduação em Medicina Tropical, Universidade de Brasília, Brasília, DF, Brazil. nnitz@unb.br.
authors.forename: Nadjar
authors.initials: N
authors.lastname: Nitz
authors.affiliation: Diretoria de Vigilância Ambiental do Distrito Federal, Brasília, DF, Brazil. laurycio@gmail.com.
authors.forename: Lucia Emilia D' Anduraim
authors.initials: LE
authors.lastname: Morales
authors.affiliation: Diretoria de Vigilância Ambiental do Distrito Federal, Brasília, DF, Brazil. laurycio@gmail.com.
authors.forename: Laurício Monteiro
authors.initials: LM
authors.lastname: Cruz
authors.affiliation: Diretoria de Vigilância Ambiental do Distrito Federal, Brasília, DF, Brazil. laurycio@gmail.com.
authors.forename: Isabele Gonçalves
authors.initials: IG
authors.lastname: Sobral
authors.affiliation: Laboratório de Pesquisa em Leishmaniose, Coleção de Leishmania, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brazil. marianacboite@gmail.com.
authors.forename: Mariana Côrtes
authors.initials: MC
authors.lastname: Boité
authors.affiliation: Laboratório de Pesquisa em Leishmaniose, Coleção de Leishmania, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brazil. ferreiragem@gmail.com.
authors.forename: Gabriel Eduardo Melim
authors.initials: GE
authors.lastname: Ferreira
authors.affiliation: Laboratório de Pesquisa em Leishmaniose, Coleção de Leishmania, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brazil. ecupoli@ioc.fiocruz.br.
authors.forename: Elisa
authors.initials: E
authors.lastname: Cupolillo
authors.affiliation: Programa de Pós-Graduação em Medicina Tropical, Universidade de Brasília, Brasília, DF, Brazil. gromero@unb.br.|Laboratório de Leishmanioses, Núcleo de Medicina Tropical, Universidade de Brasília, Brasília, DF, Brazil. gromero@unb.br.
authors.forename: Gustavo Adolfo Sierra
authors.initials: GA
authors.lastname: Romero
mesh_terms: D000818:Animals; D001096:Arachnid Vectors / Q000469:parasitology*; D004283:Dog Diseases / Q000453:epidemiology / Q000469:parasitology*; D004285:Dogs; D018314:Leishmania infantum / Q000302:isolation & purification*; D048508:Rhipicephalus sanguineus / Q000469:parasitology*; D013984:Tick Infestations / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary*