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Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs

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 Leishman...

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Autores principales: Medeiros-Silva, Viviane, Gurgel-Gonçalves, Rodrigo, Nitz, Nadjar, Morales, Lucia Emilia D’ Anduraim, Cruz, Laurício Monteiro, Sobral, Isabele Gonçalves, Boité, Mariana Côrtes, Ferreira, Gabriel Eduardo Melim, Cupolillo, Elisa, Romero, Gustavo Adolfo Sierra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4600268/
https://www.ncbi.nlm.nih.gov/pubmed/26452666
http://dx.doi.org/10.1186/s12917-015-0576-5
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author Medeiros-Silva, Viviane
Gurgel-Gonçalves, Rodrigo
Nitz, Nadjar
Morales, Lucia Emilia D’ Anduraim
Cruz, Laurício Monteiro
Sobral, Isabele Gonçalves
Boité, Mariana Côrtes
Ferreira, Gabriel Eduardo Melim
Cupolillo, Elisa
Romero, Gustavo Adolfo Sierra
author_facet Medeiros-Silva, Viviane
Gurgel-Gonçalves, Rodrigo
Nitz, Nadjar
Morales, Lucia Emilia D’ Anduraim
Cruz, Laurício Monteiro
Sobral, Isabele Gonçalves
Boité, Mariana Côrtes
Ferreira, Gabriel Eduardo Melim
Cupolillo, Elisa
Romero, Gustavo Adolfo Sierra
author_sort Medeiros-Silva, Viviane
collection PubMed
description 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. CONCLUSION: 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12917-015-0576-5) contains supplementary material, which is available to authorized users.
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spelling pubmed-46002682015-10-11 Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs Medeiros-Silva, Viviane Gurgel-Gonçalves, Rodrigo Nitz, Nadjar Morales, Lucia Emilia D’ Anduraim Cruz, Laurício Monteiro Sobral, Isabele Gonçalves Boité, Mariana Côrtes Ferreira, Gabriel Eduardo Melim Cupolillo, Elisa Romero, Gustavo Adolfo Sierra BMC Vet Res Research Article 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. CONCLUSION: 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12917-015-0576-5) contains supplementary material, which is available to authorized users. BioMed Central 2015-10-09 /pmc/articles/PMC4600268/ /pubmed/26452666 http://dx.doi.org/10.1186/s12917-015-0576-5 Text en © Medeiros-Silva et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Medeiros-Silva, Viviane
Gurgel-Gonçalves, Rodrigo
Nitz, Nadjar
Morales, Lucia Emilia D’ Anduraim
Cruz, Laurício Monteiro
Sobral, Isabele Gonçalves
Boité, Mariana Côrtes
Ferreira, Gabriel Eduardo Melim
Cupolillo, Elisa
Romero, Gustavo Adolfo Sierra
Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs
title Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs
title_full Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs
title_fullStr Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs
title_full_unstemmed Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs
title_short Successful isolation of Leishmania infantum from Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) collected from naturally infected dogs
title_sort successful isolation of leishmania infantum from rhipicephalus sanguineus sensu lato (acari: ixodidae) collected from naturally infected dogs
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4600268/
https://www.ncbi.nlm.nih.gov/pubmed/26452666
http://dx.doi.org/10.1186/s12917-015-0576-5
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