Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2015 |
| Outros Autores: | , , , , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/12281 |
Resumo: | Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil. |
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Dias, Felipe de AlmeidaGuerra, BarbaraVieira, Larissa RezendePerdomo, Hugo DiegoGandara, Ana Caroline PaivaAmaral, Raquel Juliana Vionette doVollú, Renata EstebanezGomes, Suzete Araujo OliveiraLara, Flavio AlvesSorgine, Marcos Henrique FerreiraMedei, EmilianoOliveira, Pedro Lagerblad deSalmon, Didier2015-11-19T11:52:16Z2015-11-19T11:52:16Z2015DIAS, Felipe de ALmeida; et al. Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle. PLoS Negl Trop Dis, v.9, n.10, e0004186, 23p, Oct. 2015.1935-2727https://www.arca.fiocruz.br/handle/icict/1228110.1371/journal.pntd.0004186engPlos OneMonitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Centro Nacional de Biologia Estrutural e Bioimagem—CENABIO. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Professor Paulo de Góes. Rio de Janeiro, RJ, Brasil.Universidade Federal Fluminense. Departamento de Biologia Geral. Niterói, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM). Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Centro Nacional de Biologia Estrutural e Bioimagem—CENABIO. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM). Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil.Background Here we report the monitoring of the digestive tract colonization of Rhodnius prolixus by Trypanosoma cruzi using an accurate determination of the parasite load by qPCR coupled with fluorescence and bioluminescence imaging (BLI). These complementary methods revealed critical steps necessary for the parasite population to colonize the insect gut and establish vector infection. Methodology/Principal Findings qPCR analysis of the parasite load in the insect gut showed several limitations due mainly to the presence of digestive-derived products that are thought to degrade DNA and inhibit further the PCR reaction. We developed a real-time PCR strategy targeting the T. cruzi repetitive satellite DNA sequence using as internal standard for normalization, an exogenous heterologous DNA spiked into insect samples extract, to precisely quantify the parasite load in each segment of the insect gut (anterior midgut, AM, posterior midgut, PM, and hindgut, H). Using combined fluorescence microscopy and BLI imaging as well as qPCR analysis, we showed that during their journey through the insect digestive tract, most of the parasites are lysed in the AM during the first 24 hours independently of the gut microbiota.During this short period, live parasites move through the PM to establish the onset of infection. At days 3–4 post-infection (p.i.), the parasite population begins to colonize the H to reach a climax at day 7 p.i., which is maintained during the next two weeks. Remarkably, the fluctuation of the parasite number in H remains relatively stable over the two weeks after refeeding, while the populations residing in the AM and PM increases slightly and probably constitutes the reservoirs of dividing epimastigotes. Conclusions/Significance These data show that a tuned dynamic control of the population operates in the insect gut to maintain an equilibrium between non-dividing infective trypomastigote forms and dividing epimastigote forms of the parasite, which is crucial for vector competence.Rhodnius prolixusTrypanosome Life CycleParasite LoadDigestive TractReação em Cadeia da PolimeraseCarga ParasitáriaTrato GastrointestinalTrypanosoma cruziinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; 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| dc.title.pt_BR.fl_str_mv |
Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle |
| title |
Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle |
| spellingShingle |
Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle Dias, Felipe de Almeida Rhodnius prolixus Trypanosome Life Cycle Parasite Load Digestive Tract Reação em Cadeia da Polimerase Carga Parasitária Trato Gastrointestinal Trypanosoma cruzi |
| title_short |
Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle |
| title_full |
Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle |
| title_fullStr |
Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle |
| title_full_unstemmed |
Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle |
| title_sort |
Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle |
| author |
Dias, Felipe de Almeida |
| author_facet |
Dias, Felipe de Almeida Guerra, Barbara Vieira, Larissa Rezende Perdomo, Hugo Diego Gandara, Ana Caroline Paiva Amaral, Raquel Juliana Vionette do Vollú, Renata Estebanez Gomes, Suzete Araujo Oliveira Lara, Flavio Alves Sorgine, Marcos Henrique Ferreira Medei, Emiliano Oliveira, Pedro Lagerblad de Salmon, Didier |
| author_role |
author |
| author2 |
Guerra, Barbara Vieira, Larissa Rezende Perdomo, Hugo Diego Gandara, Ana Caroline Paiva Amaral, Raquel Juliana Vionette do Vollú, Renata Estebanez Gomes, Suzete Araujo Oliveira Lara, Flavio Alves Sorgine, Marcos Henrique Ferreira Medei, Emiliano Oliveira, Pedro Lagerblad de Salmon, Didier |
| author2_role |
author author author author author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Dias, Felipe de Almeida Guerra, Barbara Vieira, Larissa Rezende Perdomo, Hugo Diego Gandara, Ana Caroline Paiva Amaral, Raquel Juliana Vionette do Vollú, Renata Estebanez Gomes, Suzete Araujo Oliveira Lara, Flavio Alves Sorgine, Marcos Henrique Ferreira Medei, Emiliano Oliveira, Pedro Lagerblad de Salmon, Didier |
| dc.subject.en.pt_BR.fl_str_mv |
Rhodnius prolixus Trypanosome Life Cycle Parasite Load Digestive Tract |
| topic |
Rhodnius prolixus Trypanosome Life Cycle Parasite Load Digestive Tract Reação em Cadeia da Polimerase Carga Parasitária Trato Gastrointestinal Trypanosoma cruzi |
| dc.subject.decs.pt_BR.fl_str_mv |
Reação em Cadeia da Polimerase Carga Parasitária Trato Gastrointestinal Trypanosoma cruzi |
| description |
Universidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Rio de Janeiro, RJ, Brasil. |
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2015 |
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2015-11-19T11:52:16Z |
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2015-11-19T11:52:16Z |
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2015 |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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DIAS, Felipe de ALmeida; et al. Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle. PLoS Negl Trop Dis, v.9, n.10, e0004186, 23p, Oct. 2015. |
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https://www.arca.fiocruz.br/handle/icict/12281 |
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1935-2727 |
| dc.identifier.doi.none.fl_str_mv |
10.1371/journal.pntd.0004186 |
| identifier_str_mv |
DIAS, Felipe de ALmeida; et al. Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle. PLoS Negl Trop Dis, v.9, n.10, e0004186, 23p, Oct. 2015. 1935-2727 10.1371/journal.pntd.0004186 |
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eng |
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Plos One |
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Plos One |
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