Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicos
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/18028 |
Resumo: | Pythium insidiosum is an aquatic oomycete that causes pythiosis, a pyogranulomatous disease of difficult treatment that infects animals and humans. In Brazil, the disease is often described in horses in the regions of the Pantanal and Rio Grande do Sul. Thailand is an endemic country for human pythiosis, where the disease has important morbidity and mortality rates. Pythiosis presents a rapid progression, there is no standard treatment available and unresponsive cases to chemotherapy are observed. P. insidiosum has no ergosterol in the cellular membrane, and this component is the main target of antifungal drugs currently available. This study aimed to analyze the phylogenetic and phylogeographic characteristics of Brazilian and Thai isolates of P. insidiosum in order to better understand the evolution of this oomycete, as well as to characterize the susceptibility of P. insidiosum against metal compounds in search of new therapeutic alternatives. This study included 16 partial DNA sequences of Exo-1,3-ß-glucanase gene (Exo1) of Brazilian isolates of P. insidiosum, 23 sequences of Thai isolates and 2 standard strains. Phylogenetic analyses employed the methods of maximum parsimony (MP), neighbor-joining (NJ), maximum likelihood (ML), and Bayesian analysis (BA). The phylogeographic analyses involved evaluation of the nucleotide diversity, of polymorphism, of molecular variance (SAMOVA and AMOVA), the degree of genetic diversity (FST) and neutrality tests, which were performed with 31 exo1 the DNA sequences of P. insidiosum with known geographical origin. exo1 demonstrated to be a good marker for phylogenetic and phylogeographic studies. A tree consensus of the four methods was obtained, and observed three distinct groups consistent with previous analyzes with other molecular markers. In the phylogeographic analyses, the Thai isolates display a high diversity, confirming its earlier origin in relation to Brazilian isolates. Structuring patterns were observed among Brazilian isolates, suggesting a recent expansion of these and indicates the need for further studies with larger sample size of these regions. In the evaluation of in vitro susceptibility of P. insidiosum (n = 23) to metallic compounds containing cadmium, lead, copper, manganese, and zinc were performed using microdilution broth test based on document M38-A2. The minimum inhibitory and fungicidal concentrations were established for all isolates. Copper acetate and cadmium acetate showed minimal inhibitory concentration range of 4-64 mg/mL and 16-256 mg/mL, respectively. These results suggest that copper and cadmium can inhibit the growth of P. insidiosum, emphasizing the most inhibitory activity of copper acetate. Furthermore, this study suggests that compounds of copper and/or cadmium can be used in future research to develop effective drugs against P. insidiosum. |
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Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicosPhylogenetic and phylogeographic analysis of Pythium insidiosum isolates with the genetic marker Exo1 and susceptibility to metal compoundsOomicetoP. insidisoumFilogeniaFilogeografiaSusceptibilidadeMetaisOomycetePhylogenyPhylogeographySusceptibilityMetalsCNPQ::CIENCIAS DA SAUDE::FARMACIAPythium insidiosum is an aquatic oomycete that causes pythiosis, a pyogranulomatous disease of difficult treatment that infects animals and humans. In Brazil, the disease is often described in horses in the regions of the Pantanal and Rio Grande do Sul. Thailand is an endemic country for human pythiosis, where the disease has important morbidity and mortality rates. Pythiosis presents a rapid progression, there is no standard treatment available and unresponsive cases to chemotherapy are observed. P. insidiosum has no ergosterol in the cellular membrane, and this component is the main target of antifungal drugs currently available. This study aimed to analyze the phylogenetic and phylogeographic characteristics of Brazilian and Thai isolates of P. insidiosum in order to better understand the evolution of this oomycete, as well as to characterize the susceptibility of P. insidiosum against metal compounds in search of new therapeutic alternatives. This study included 16 partial DNA sequences of Exo-1,3-ß-glucanase gene (Exo1) of Brazilian isolates of P. insidiosum, 23 sequences of Thai isolates and 2 standard strains. Phylogenetic analyses employed the methods of maximum parsimony (MP), neighbor-joining (NJ), maximum likelihood (ML), and Bayesian analysis (BA). The phylogeographic analyses involved evaluation of the nucleotide diversity, of polymorphism, of molecular variance (SAMOVA and AMOVA), the degree of genetic diversity (FST) and neutrality tests, which were performed with 31 exo1 the DNA sequences of P. insidiosum with known geographical origin. exo1 demonstrated to be a good marker for phylogenetic and phylogeographic studies. A tree consensus of the four methods was obtained, and observed three distinct groups consistent with previous analyzes with other molecular markers. In the phylogeographic analyses, the Thai isolates display a high diversity, confirming its earlier origin in relation to Brazilian isolates. Structuring patterns were observed among Brazilian isolates, suggesting a recent expansion of these and indicates the need for further studies with larger sample size of these regions. In the evaluation of in vitro susceptibility of P. insidiosum (n = 23) to metallic compounds containing cadmium, lead, copper, manganese, and zinc were performed using microdilution broth test based on document M38-A2. The minimum inhibitory and fungicidal concentrations were established for all isolates. Copper acetate and cadmium acetate showed minimal inhibitory concentration range of 4-64 mg/mL and 16-256 mg/mL, respectively. These results suggest that copper and cadmium can inhibit the growth of P. insidiosum, emphasizing the most inhibitory activity of copper acetate. Furthermore, this study suggests that compounds of copper and/or cadmium can be used in future research to develop effective drugs against P. insidiosum.Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul, FAPERGS, Brasil.Pythium insidiosum é o oomiceto aquático causador da pitiose, uma doença piogranulomatosa de difícil tratamento que acomete os animais e humanos. No Brasil, a doença é frequentemente descrita em equinos nas regiões do Pantanal Mato-grossense e no Rio Grande do Sul. A Tailândia é um país endêmico para a pitiose humana, onde a doença apresenta importantes níveis de morbidade e mortalidade. A pitiose apresenta uma progressão rápida e não há um tratamento padrão disponível, ocorrendo casos não responsivos aos quimioterápicos. P. insidiosum não possui ergosterol na membrana celular, e este componente é o principal alvo dos antifúngicos disponíveis atualmente. Este trabalho buscou analisar as características filogenéticas e filogeográficas de isolados brasileiros e tailandeses de P. insidiosum a fim de melhor compreender a evolução deste oomiceto, bem como caracterizar a susceptibilidade de P. insidiosum frente a compostos metálicos em busca de novas alternativas terapêuticas. Este estudo incluiu 16 sequências parciais de DNA da região Exo-1,3-ß-Glucanase (exo1) de isolados brasileiros de P. insidiosum, 23 sequências de isolados tailandeses e 2 cepas-padrão. As análises filogenéticas empregaram as metodologias de máxima parcimônia (MP), Neighbor-joining (NJ), máxima verossimilhança (ML) e análise Bayesiana (BA). As análises filogeográficas envolveram a avaliação da diversidade nucleotídica, do polimorfismo, da variância molecular (SAMOVA e AMOVA), do grau de divergência genética (FST) e testes de neutralidade, os quais foram realizados com 31 das sequências de DNA exo1 de P. insidiosum com origem geográfica conhecida. exo1 provou ser um bom marcador para estudos filogenéticos e filogeográficos. Foi obtida uma árvore consenso dos quatro métodos utilizados, onde observou-se três grupos distintos compatíveis com análises anteriores com outros marcadores moleculares. Nas análises filogeográficas, os isolados tailandeses apresentaram grande diversidade, confirmando sua origem mais antiga em relação aos isolados brasileiros. Foram observados padrões de estruturação entre os isolados brasileiros, sugerindo uma expansão recente destes e apontando a necessidade de novos estudos com maior número de amostras destas regiões. Na avaliação da susceptibilidade in vitro de P. insidiosum (n=23) aos compostos metálicos contendo cádmio, chumbo, cobre, manganês e zinco foram realizados o teste de microdiluição em caldo com base no documento M38-A2. As concentrações inibitórias e fungicidas mínimas foram estabelecidas para todos os isolados. Acetato de cobre e acetato de cádmio apresentaram concentração inibitória mínima variando 4-64 μg/mL e 16-256 μg/mL, respectivamente. Estes resultados sugerem que o cobre e o cádmio podem inibir o crescimento de P. insidiosum, destacando a maior atividade inibidora de acetato de cobre. Além disso, este estudo sugere que compostos de cobre e/ou cádmio podem ser utilizados em futuras pesquisas para formular fármacos eficazes contra P. insidiosum.Universidade Federal de Santa MariaBrasilFarmacologiaUFSMPrograma de Pós-Graduação em FarmacologiaCentro de Ciências da SaúdeSanturio, Janio Moraishttp://lattes.cnpq.br/6316012260769979Zanette, Régis Adrielhttp://lattes.cnpq.br/3166371422900794Loreto, Érico Silva dehttp://lattes.cnpq.br/5475233864057995Ribeiro, Tatiana Corrêa2019-08-26T22:10:56Z2019-08-26T22:10:56Z2016-02-26info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/18028porAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2019-08-27T06:01:33Zoai:repositorio.ufsm.br:1/18028Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2019-08-27T06:01:33Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicos Phylogenetic and phylogeographic analysis of Pythium insidiosum isolates with the genetic marker Exo1 and susceptibility to metal compounds |
title |
Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicos |
spellingShingle |
Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicos Ribeiro, Tatiana Corrêa Oomiceto P. insidisoum Filogenia Filogeografia Susceptibilidade Metais Oomycete Phylogeny Phylogeography Susceptibility Metals CNPQ::CIENCIAS DA SAUDE::FARMACIA |
title_short |
Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicos |
title_full |
Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicos |
title_fullStr |
Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicos |
title_full_unstemmed |
Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicos |
title_sort |
Análise filogenética e filogeográfica de isolados de Pythium insidiosum através do marcador genético Exo1 e suscetibilidade aos compostos metálicos |
author |
Ribeiro, Tatiana Corrêa |
author_facet |
Ribeiro, Tatiana Corrêa |
author_role |
author |
dc.contributor.none.fl_str_mv |
Santurio, Janio Morais http://lattes.cnpq.br/6316012260769979 Zanette, Régis Adriel http://lattes.cnpq.br/3166371422900794 Loreto, Érico Silva de http://lattes.cnpq.br/5475233864057995 |
dc.contributor.author.fl_str_mv |
Ribeiro, Tatiana Corrêa |
dc.subject.por.fl_str_mv |
Oomiceto P. insidisoum Filogenia Filogeografia Susceptibilidade Metais Oomycete Phylogeny Phylogeography Susceptibility Metals CNPQ::CIENCIAS DA SAUDE::FARMACIA |
topic |
Oomiceto P. insidisoum Filogenia Filogeografia Susceptibilidade Metais Oomycete Phylogeny Phylogeography Susceptibility Metals CNPQ::CIENCIAS DA SAUDE::FARMACIA |
description |
Pythium insidiosum is an aquatic oomycete that causes pythiosis, a pyogranulomatous disease of difficult treatment that infects animals and humans. In Brazil, the disease is often described in horses in the regions of the Pantanal and Rio Grande do Sul. Thailand is an endemic country for human pythiosis, where the disease has important morbidity and mortality rates. Pythiosis presents a rapid progression, there is no standard treatment available and unresponsive cases to chemotherapy are observed. P. insidiosum has no ergosterol in the cellular membrane, and this component is the main target of antifungal drugs currently available. This study aimed to analyze the phylogenetic and phylogeographic characteristics of Brazilian and Thai isolates of P. insidiosum in order to better understand the evolution of this oomycete, as well as to characterize the susceptibility of P. insidiosum against metal compounds in search of new therapeutic alternatives. This study included 16 partial DNA sequences of Exo-1,3-ß-glucanase gene (Exo1) of Brazilian isolates of P. insidiosum, 23 sequences of Thai isolates and 2 standard strains. Phylogenetic analyses employed the methods of maximum parsimony (MP), neighbor-joining (NJ), maximum likelihood (ML), and Bayesian analysis (BA). The phylogeographic analyses involved evaluation of the nucleotide diversity, of polymorphism, of molecular variance (SAMOVA and AMOVA), the degree of genetic diversity (FST) and neutrality tests, which were performed with 31 exo1 the DNA sequences of P. insidiosum with known geographical origin. exo1 demonstrated to be a good marker for phylogenetic and phylogeographic studies. A tree consensus of the four methods was obtained, and observed three distinct groups consistent with previous analyzes with other molecular markers. In the phylogeographic analyses, the Thai isolates display a high diversity, confirming its earlier origin in relation to Brazilian isolates. Structuring patterns were observed among Brazilian isolates, suggesting a recent expansion of these and indicates the need for further studies with larger sample size of these regions. In the evaluation of in vitro susceptibility of P. insidiosum (n = 23) to metallic compounds containing cadmium, lead, copper, manganese, and zinc were performed using microdilution broth test based on document M38-A2. The minimum inhibitory and fungicidal concentrations were established for all isolates. Copper acetate and cadmium acetate showed minimal inhibitory concentration range of 4-64 mg/mL and 16-256 mg/mL, respectively. These results suggest that copper and cadmium can inhibit the growth of P. insidiosum, emphasizing the most inhibitory activity of copper acetate. Furthermore, this study suggests that compounds of copper and/or cadmium can be used in future research to develop effective drugs against P. insidiosum. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-02-26 2019-08-26T22:10:56Z 2019-08-26T22:10:56Z |
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.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/18028 |
url |
http://repositorio.ufsm.br/handle/1/18028 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Farmacologia UFSM Programa de Pós-Graduação em Farmacologia Centro de Ciências da Saúde |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Farmacologia UFSM Programa de Pós-Graduação em Farmacologia Centro de Ciências da Saúde |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
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1805922146119057408 |