Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados

Detalhes bibliográficos
Autor(a) principal: Pires, Marize Quinhones
Data de Publicação: 2010
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRRJ
Texto Completo: https://tede.ufrrj.br/jspui/handle/jspui/4029
Resumo: Visceral leishmaniasis is a potentially zoonotic disease with a broad geographical distribution. Dogs play a crucial role in its epidemiology, being considered as important domestic reservoirs for Leishmania (Leishmania) chagasi in the Americas. In these dogs, L. (L.) chagasi presents an extensive tissue distribution, being found in organs such as the spleen, the liver, lymph nodes, skin and bone marrow. Despite diagnostic investigation is usually performed in the skin, the clonal variability of L. (L.) chagasi subpopulations in the skin (and other affected organs) is not well understood. The main objective of this study was to: i) perform specific molecular diagnosis of leishmaniasis through PCR, and ii) assess intraspecific genetic variability of distinct L. (L.) chagasi strains characterized in different host tissues by two molecular techniques (LSSP-PCR and RFLP-PCR). Fragments of normal skin, cutaneous lesions, spleen, liver and lymph nodes obtained from naturally infected dogs living in the state of Rio de Janeiro were analyzed. One hundred six tissue fragments obtained from 17 dogs were studied. Through PCR, 66 (62.26%) of the fragments proved positive utilizing specific primers D1/D2 to species belonging to the L. donovani complex. Cervical and popliteous lymph nodes presented the highest rate of positivity (100%, 17/17). Spleen and normal skin presented detection rates of 76.47% (13/17) and 70.59% (12/17), respectively. RFLP-PCR was performed in order to confirm the presence of L. (V.) braziliensis and L. (L.) chagasi DNA in cutaneous lesions and in internal organs, respectively, detecting two cases of co-infection with both parasites. Through LSSP-PCR it was possible to demonstrate that L. (L.) chagasi infection courses with parasitic spread and different tissue tropism of genetically distinct subpopulations of the parasite. Nevertheless, it was not possible to correlate specific genetic profiles with distinct clinical outcomes. The present study contributed to understand the dynamics of L. (L.) chagasi infection, concerning tissue tropism and the role of distinct parasitic subpopulations of L. (L.) chagasi. In addition, it was possible to assess the genetic variability of some strains circulating in dogs in the studied area.
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spelling Bittencourt, V?nia Rita Elias Pinheiro505.198.676-53http://lattes.cnpq.br/3888832724995864Pacheco, Raquel da SilvaMadeira, Maria de F?timaBonfim, Teresa Cristina Bergamo018.495.317-06http://lattes.cnpq.br/1972618154471192Pires, Marize Quinhones2020-10-20T22:16:36Z2010-06-28PIRES, Marize Quinhones. Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados. 2010. 46 f. Disserta??o (Programa Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.https://tede.ufrrj.br/jspui/handle/jspui/4029Visceral leishmaniasis is a potentially zoonotic disease with a broad geographical distribution. Dogs play a crucial role in its epidemiology, being considered as important domestic reservoirs for Leishmania (Leishmania) chagasi in the Americas. In these dogs, L. (L.) chagasi presents an extensive tissue distribution, being found in organs such as the spleen, the liver, lymph nodes, skin and bone marrow. Despite diagnostic investigation is usually performed in the skin, the clonal variability of L. (L.) chagasi subpopulations in the skin (and other affected organs) is not well understood. The main objective of this study was to: i) perform specific molecular diagnosis of leishmaniasis through PCR, and ii) assess intraspecific genetic variability of distinct L. (L.) chagasi strains characterized in different host tissues by two molecular techniques (LSSP-PCR and RFLP-PCR). Fragments of normal skin, cutaneous lesions, spleen, liver and lymph nodes obtained from naturally infected dogs living in the state of Rio de Janeiro were analyzed. One hundred six tissue fragments obtained from 17 dogs were studied. Through PCR, 66 (62.26%) of the fragments proved positive utilizing specific primers D1/D2 to species belonging to the L. donovani complex. Cervical and popliteous lymph nodes presented the highest rate of positivity (100%, 17/17). Spleen and normal skin presented detection rates of 76.47% (13/17) and 70.59% (12/17), respectively. RFLP-PCR was performed in order to confirm the presence of L. (V.) braziliensis and L. (L.) chagasi DNA in cutaneous lesions and in internal organs, respectively, detecting two cases of co-infection with both parasites. Through LSSP-PCR it was possible to demonstrate that L. (L.) chagasi infection courses with parasitic spread and different tissue tropism of genetically distinct subpopulations of the parasite. Nevertheless, it was not possible to correlate specific genetic profiles with distinct clinical outcomes. The present study contributed to understand the dynamics of L. (L.) chagasi infection, concerning tissue tropism and the role of distinct parasitic subpopulations of L. (L.) chagasi. In addition, it was possible to assess the genetic variability of some strains circulating in dogs in the studied area.Na leishmaniose visceral (LV), uma doen?a de potencial zoon?tico e de ampla distribui??o, o c?o possui um papel bem sedimentado, sendo considerado um importante reservat?rio dom?stico de Leishmania (Leishmania) chagasi nas Am?ricas. Nestes animais, a distribui??o do parasita pode ser extensa, alcan?ando ?rg?os como ba?o, f?gado, lifonodos, pele e medula ?ssea. ? o parasitismo de pele ?ntegra que faz do c?o um elo importante na cadeia epidemiol?gica da doen?a. Embora a pele seja um s?tio de investiga??o diagn?stica, n?o se conhece a variabilidade clonal da distribui??o de subpopula??es de L.(L.) chagasi neste e nos demais ?rg?os nos c?es infectados. O principal objetivo do presente trabalho foi utilizar o ensaio da PCR para o diagn?stico espec?fico da infec??o e, investigar atrav?s de t?cnicas moleculares (LSSP-PCR e RFLP-PCR) a variabilidade gen?tica intraespec?fica de L.(L.) chagasi presente em fragmentos de pele ?ntegra, les?o cut?nea, ba?o, f?gado e linfonodos de c?es naturalmente infectados do Munic?pio do Rio de Janeiro. De um total de 17 c?es, 106 fragmentos teciduais foram analisados. Destes, 66 (62,26%) foram PCR positivos com os iniciadores D1/D2 espec?ficos para as esp?cies do complexo L. donovani.. O maior percentual de detec??o foi observado em fragmentos de linfonodos cervical (17/17, 100%) e popl?teo (17/17, 100%), seguido do ba?o com o percentual de 76,47% (13/17) e, em pele ?ntegra com 12 biopsias positivas em 17 (70,59%). A t?cnica de RFLP-PCR, associada aos ensaios de PCR, foi escolhida para confirmar a presen?a de DNA de L. (V.) braziliensis em les?es cut?neas e de L. (L.) chagasi em fragmentos de ?rg?os internos de dois c?es, confirmando dois casos caninos de coinfec??o no Munic?pio do Rio de Janeiro. Atrav?s da t?cnica de LSSP-PCR foi poss?vel evidenciar que a infec??o canina por L. (L.) chagasi pode manisfestar-se com dissemina??o parasit?ria e tropismo tissular por popula??es do parasita geneticamente diferenciadas. N?o foi poss?vel, no entanto, observar associa??o entre determinado perfil gen?tico e o estado cl?nico do animal. O presente estudo contribuiu para um melhor entendimento da din?mica da infec??o na LV canina no que diz respeito ao tropismo diferenciado e ao envolvimento de subpopula??es parasit?rias de L. (L.) chagasi. Al?m disso, o estudo permitiu rastrear a variabilidade gen?tica de algumas cepas circulantes na popula??o canina das ?reas investigadas.Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2020-10-20T22:16:36Z No. of bitstreams: 1 2010 - Marize Quinhones Pires.pdf: 2876106 bytes, checksum: 2d526905fb58579977acc2c8b7937689 (MD5)Made available in DSpace on 2020-10-20T22:16:36Z (GMT). No. of bitstreams: 1 2010 - Marize Quinhones Pires.pdf: 2876106 bytes, checksum: 2d526905fb58579977acc2c8b7937689 (MD5) Previous issue date: 2010-06-28Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico, CNPq, BrasilFunda??o de Amparo a Pesquisa do Estado do Rio de Janeiroapplication/pdfhttps://tede.ufrrj.br/retrieve/62535/2010%20-%20Marize%20Quinhones%20Pires.pdf.jpgporUniversidade Federal Rural do Rio de JaneiroPrograma de P?s-Gradua??o em Ci?ncias Veterin?riasUFRRJBrasilInstituto de Veterin?riaABRANCHES, P.; SILVA PEREIRA, M. C. D.; CONCEI??O-SILVA, F. M.; SANTANAGOMES, G. M.; JANS, J. G. 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dc.title.por.fl_str_mv Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados
dc.title.alternative.eng.fl_str_mv PCR diagnosis and study of the genetic variability of Leishmania (Leishmania) chagasi in distinct anatomic sites of naturally infected dogs.
title Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados
spellingShingle Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados
Pires, Marize Quinhones
leishmaniose visceral canina
LSSP-PCR
RFLP-PCR
canine visceral leishmaniasis
LSSSP-PCR
RFLP-PCR
Medicina Veterin?ria
title_short Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados
title_full Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados
title_fullStr Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados
title_full_unstemmed Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados
title_sort Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados
author Pires, Marize Quinhones
author_facet Pires, Marize Quinhones
author_role author
dc.contributor.advisor1.fl_str_mv Bittencourt, V?nia Rita Elias Pinheiro
dc.contributor.advisor1ID.fl_str_mv 505.198.676-53
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/3888832724995864
dc.contributor.advisor-co1.fl_str_mv Pacheco, Raquel da Silva
dc.contributor.referee1.fl_str_mv Madeira, Maria de F?tima
dc.contributor.referee2.fl_str_mv Bonfim, Teresa Cristina Bergamo
dc.contributor.authorID.fl_str_mv 018.495.317-06
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/1972618154471192
dc.contributor.author.fl_str_mv Pires, Marize Quinhones
contributor_str_mv Bittencourt, V?nia Rita Elias Pinheiro
Pacheco, Raquel da Silva
Madeira, Maria de F?tima
Bonfim, Teresa Cristina Bergamo
dc.subject.por.fl_str_mv leishmaniose visceral canina
LSSP-PCR
RFLP-PCR
topic leishmaniose visceral canina
LSSP-PCR
RFLP-PCR
canine visceral leishmaniasis
LSSSP-PCR
RFLP-PCR
Medicina Veterin?ria
dc.subject.eng.fl_str_mv canine visceral leishmaniasis
LSSSP-PCR
RFLP-PCR
dc.subject.cnpq.fl_str_mv Medicina Veterin?ria
description Visceral leishmaniasis is a potentially zoonotic disease with a broad geographical distribution. Dogs play a crucial role in its epidemiology, being considered as important domestic reservoirs for Leishmania (Leishmania) chagasi in the Americas. In these dogs, L. (L.) chagasi presents an extensive tissue distribution, being found in organs such as the spleen, the liver, lymph nodes, skin and bone marrow. Despite diagnostic investigation is usually performed in the skin, the clonal variability of L. (L.) chagasi subpopulations in the skin (and other affected organs) is not well understood. The main objective of this study was to: i) perform specific molecular diagnosis of leishmaniasis through PCR, and ii) assess intraspecific genetic variability of distinct L. (L.) chagasi strains characterized in different host tissues by two molecular techniques (LSSP-PCR and RFLP-PCR). Fragments of normal skin, cutaneous lesions, spleen, liver and lymph nodes obtained from naturally infected dogs living in the state of Rio de Janeiro were analyzed. One hundred six tissue fragments obtained from 17 dogs were studied. Through PCR, 66 (62.26%) of the fragments proved positive utilizing specific primers D1/D2 to species belonging to the L. donovani complex. Cervical and popliteous lymph nodes presented the highest rate of positivity (100%, 17/17). Spleen and normal skin presented detection rates of 76.47% (13/17) and 70.59% (12/17), respectively. RFLP-PCR was performed in order to confirm the presence of L. (V.) braziliensis and L. (L.) chagasi DNA in cutaneous lesions and in internal organs, respectively, detecting two cases of co-infection with both parasites. Through LSSP-PCR it was possible to demonstrate that L. (L.) chagasi infection courses with parasitic spread and different tissue tropism of genetically distinct subpopulations of the parasite. Nevertheless, it was not possible to correlate specific genetic profiles with distinct clinical outcomes. The present study contributed to understand the dynamics of L. (L.) chagasi infection, concerning tissue tropism and the role of distinct parasitic subpopulations of L. (L.) chagasi. In addition, it was possible to assess the genetic variability of some strains circulating in dogs in the studied area.
publishDate 2010
dc.date.issued.fl_str_mv 2010-06-28
dc.date.accessioned.fl_str_mv 2020-10-20T22:16:36Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv PIRES, Marize Quinhones. Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados. 2010. 46 f. Disserta??o (Programa Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.
dc.identifier.uri.fl_str_mv https://tede.ufrrj.br/jspui/handle/jspui/4029
identifier_str_mv PIRES, Marize Quinhones. Diagn?stico por PCR e estudo da variabilidade gen?tica de Leishmania (Leishmania) chagasi em diferentes s?tios anat?micos de c?es naturalmente infectados. 2010. 46 f. Disserta??o (Programa Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.
url https://tede.ufrrj.br/jspui/handle/jspui/4029
dc.language.iso.fl_str_mv por
language por
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L.; TAFURI, W. L.; BARBOSA, A.J. A.; MICHALICK, M. S.; GENERO, O.; FRAN?A-SILVA, J. C., MAYRIK, W.; NASCIMENTO, E. Histopathology and immunocytochemical study of type 3 and type 4 complement receptors in the liver and splleen of dogs naturally and
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