Imunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosum
Autor(a) principal: | |
---|---|
Data de Publicação: | 2011 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional Manancial UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/8953 |
Resumo: | The Pythium insidiosum is an aquatic oomycete that is the causative agent of pythiosis, an important disease in humans and animals that is prevalent in tropical and subtropical areas. In Brazil it was first reported in 1974, then several cases this disease has been reported throughout the country. The failure in the antifungal therapy combined with cases unresponsive to immunotherapy existing, lead to molecular studies in order to investigate possible genetic variations among Brazilian isolates, and this may be able to contribute on the research for the improvement of immunotherapy available. Thus, this study aimed to evaluate the genetic diversity of thirty P. insidiosum isolates from different regions of Brazil, and compare all of them with isolates from Thailand, one isolate from Central America and another isolate from North America, by sequencing and the analysis of genomic DNA regions corresponding to cytochrome c oxidase (COX II) and ITS1, 5.8S rRNA and ITS2 rDNA (ITS). The analyses of nucleotide sequences of both regions were carried out, individually and in combination, using the following methods: Maximum parsimony (MP); Neighbor-joining (NJ); Maximum likelihood (ML); and Bayesian analysis (BA). Our data demonstrated all of P. insidiosum isolates as monophyletic in relation to the other Pythium species. Analises of COX II gene sequences subdivided P. insidiosum isolates into three groups, whose arrangement provides the Thai isolates as paraphyletic in relation to the Brazilian isolates. The molecular analyses performed in this study suggest an evolutionary proximity among all of American isolates, including the Brazilian, from Costa Rica and from the United States, that were grouped in a single group. COX II gene network analysis showed signs of a recent expansion of P. insidosum isolates, probably originated from the Asiatic to America continent. By analysis of COX II gene demonstrated the highest levels of genetic variability among P. insidiosum isolates studied in here, additionally, the highest levels of phylogenetic information were shown, when it was compared to the ITS region analysis. Nevertheless, both genetic markers selected for this study proved to be entirely congruent in the phylogenetic relations among Brazilian isolates of P. insidiosum, since clustered all of them into a single monophyletic group which did not shown to have genetic variability. The results indicate that the strain used in the production of the immunotherapic - Pitium-Vac ® - is representative of the Brazilian isolates of P. insidiosum. |
id |
UFSM-20_0d85a41a5cd6d51c2dd94658b918feda |
---|---|
oai_identifier_str |
oai:repositorio.ufsm.br:1/8953 |
network_acronym_str |
UFSM-20 |
network_name_str |
Repositório Institucional Manancial UFSM |
repository_id_str |
3913 |
spelling |
2012-05-082012-05-082011-07-15AZEVEDO, Maria Isabel de. IMMUNETHERAPY AGAINST PYTHIOSIS: MOLECULAR CHARACTERIZATION OF BRAZILIAN ISOLATES OF Pythium insidiosum. 2011. 87 f. Dissertação (Mestrado em Farmácia) - Universidade Federal de Santa Maria, Santa Maria, 2011.http://repositorio.ufsm.br/handle/1/8953The Pythium insidiosum is an aquatic oomycete that is the causative agent of pythiosis, an important disease in humans and animals that is prevalent in tropical and subtropical areas. In Brazil it was first reported in 1974, then several cases this disease has been reported throughout the country. The failure in the antifungal therapy combined with cases unresponsive to immunotherapy existing, lead to molecular studies in order to investigate possible genetic variations among Brazilian isolates, and this may be able to contribute on the research for the improvement of immunotherapy available. Thus, this study aimed to evaluate the genetic diversity of thirty P. insidiosum isolates from different regions of Brazil, and compare all of them with isolates from Thailand, one isolate from Central America and another isolate from North America, by sequencing and the analysis of genomic DNA regions corresponding to cytochrome c oxidase (COX II) and ITS1, 5.8S rRNA and ITS2 rDNA (ITS). The analyses of nucleotide sequences of both regions were carried out, individually and in combination, using the following methods: Maximum parsimony (MP); Neighbor-joining (NJ); Maximum likelihood (ML); and Bayesian analysis (BA). Our data demonstrated all of P. insidiosum isolates as monophyletic in relation to the other Pythium species. Analises of COX II gene sequences subdivided P. insidiosum isolates into three groups, whose arrangement provides the Thai isolates as paraphyletic in relation to the Brazilian isolates. The molecular analyses performed in this study suggest an evolutionary proximity among all of American isolates, including the Brazilian, from Costa Rica and from the United States, that were grouped in a single group. COX II gene network analysis showed signs of a recent expansion of P. insidosum isolates, probably originated from the Asiatic to America continent. By analysis of COX II gene demonstrated the highest levels of genetic variability among P. insidiosum isolates studied in here, additionally, the highest levels of phylogenetic information were shown, when it was compared to the ITS region analysis. Nevertheless, both genetic markers selected for this study proved to be entirely congruent in the phylogenetic relations among Brazilian isolates of P. insidiosum, since clustered all of them into a single monophyletic group which did not shown to have genetic variability. The results indicate that the strain used in the production of the immunotherapic - Pitium-Vac ® - is representative of the Brazilian isolates of P. insidiosum.O Pythium insidiosum é um oomiceto aquático causador da pitiose, uma importante enfermidade em humanos e animais, sendo prevalente em áreas tropicais e subtropicais. No Brasil foi relatada pela primeira vez em 1974. Desde então vários casos desta enfermidade tem sido descritos por todo o país. Os insucessos de terapias antifúngicas aliados aos casos não responsivos à imunoterapia existente impulsionam estudos moleculares a fim de investigar possíveis variações genéticas entre os isolados brasileiros de forma a contribuir nas pesquisas para a melhoria do imunoterápico disponível. Assim, o presente estudo teve por objetivo avaliar a diversidade genética de 30 isolados de P. insidiosum provenientes de diferentes regiões do Brasil, bem como compará-los com isolados tailandeses, um isolado da América Central e outro isolado da América do Norte através do sequenciamento e das análises das regiões do DNA genômico correspondentes à citocromo c oxidase (COX II) e ITS1, 5.8S rRNA e ITS2 do rDNA (ITS). As análises das sequências de nucleotídeos de ambas as regiões foram realizadas, individualmente e em combinação, utilizando as seguintes metodologias: máxima parcimônia (Maximum parsimony, MP); Neighbor-joining (NJ); máxima verossimilhança (Maximum likelihood, ML); e análise Bayesiana (Bayesian analysis, BA). Os dados demonstraram que todos os isolados de P. insidiosum são monofiléticos em relação às outras espécies de Pythium. A análise das sequências do gene COX II subdividiu os isolados de P. insidiosum em três grupos, cuja disposição demonstra os isolados tailandeses como parafilético em relação aos isolados brasileiros. As análises moleculares realizadas neste estudo sugerem uma proximidade evolutiva entre todos os isolados americanos, incluindo os brasileiros, da Costa Rica e dos Estados Unidos, os quais foram agrupados juntos em um único grupo. Na análise de network do gene COX II os resultados apresentaram sinais de uma recente expansão dos isolados de P. insidosum para a América, provavelmente oriundos do continente asiático. Pela análise do gene COX II foi possível evidenciar os maiores níveis de variabilidade genética entre os isolados de P. insidiosum avaliados, além disso, foram demonstrados os maiores níveis de informação filogenética, quando comparada à análise da região ITS. Contudo, os dois marcadores genéticos selecionados para este estudo revelaram-se inteiramente congruentes nas relações filogenéticas entre os isolados brasileiros de P. insidiosum, reunindo-os em um único grupo monofilético o qual não demonstrou haver variabilidade genética. Os resultados obtidos indicam que a cepa utilizada na produção do imunoterápico Pitium-Vac ® é representativa dos isolados brasileiros de P. insidiosum.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de Santa MariaPrograma de Pós-Graduação em FarmacologiaUFSMBRFarmacologiaPitioseCaracterização molecularFilogeniaImunoterapiaPythium insidiosumPythiosisPhylogenyMolecular characterizationImmunotherapyCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIAImunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosumImmunetherapy against pythiosis: molecular characterization of brazilian isolates of Pythium insidiosuminfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisSanturio, Janio Moraishttp://lattes.cnpq.br/6316012260769979Leal, Daniela Bitencourt Rosahttp://lattes.cnpq.br/3639683273462361Loreto, érico Silva dehttp://lattes.cnpq.br/5475233864057995http://lattes.cnpq.br/7516243030317280Azevedo, Maria Isabel de20100000000040050030050050006a1b29c-fde0-4a69-8097-2a9d6a9d964981087730-2239-428b-ad21-591665a7aecf2dbfa8b1-3511-410e-8203-4bb0cf639ac8ea251119-5531-432f-bf54-8c8111248727info:eu-repo/semantics/openAccessreponame:Repositório Institucional Manancial UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALAZEVEDO, MARIA ISABEL DE.pdfapplication/pdf657625http://repositorio.ufsm.br/bitstream/1/8953/1/AZEVEDO%2c%20MARIA%20ISABEL%20DE.pdf82fc06a0117d4629043b9d8ba576cd27MD51TEXTAZEVEDO, MARIA ISABEL DE.pdf.txtAZEVEDO, MARIA ISABEL DE.pdf.txtExtracted texttext/plain148254http://repositorio.ufsm.br/bitstream/1/8953/2/AZEVEDO%2c%20MARIA%20ISABEL%20DE.pdf.txta7895a29b572186209b75fb5bb8b2da3MD52THUMBNAILAZEVEDO, MARIA ISABEL DE.pdf.jpgAZEVEDO, MARIA ISABEL DE.pdf.jpgIM Thumbnailimage/jpeg4702http://repositorio.ufsm.br/bitstream/1/8953/3/AZEVEDO%2c%20MARIA%20ISABEL%20DE.pdf.jpgd3be45d5629d80e99de9ea1446cc92a2MD531/89532021-10-22 10:57:30.753oai:repositorio.ufsm.br:1/8953Repositório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestouvidoria@ufsm.bropendoar:39132021-10-22T13:57:30Repositório Institucional Manancial UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.por.fl_str_mv |
Imunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosum |
dc.title.alternative.eng.fl_str_mv |
Immunetherapy against pythiosis: molecular characterization of brazilian isolates of Pythium insidiosum |
title |
Imunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosum |
spellingShingle |
Imunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosum Azevedo, Maria Isabel de Pitiose Caracterização molecular Filogenia Imunoterapia Pythium insidiosum Pythiosis Phylogeny Molecular characterization Immunotherapy CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
title_short |
Imunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosum |
title_full |
Imunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosum |
title_fullStr |
Imunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosum |
title_full_unstemmed |
Imunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosum |
title_sort |
Imunoterapia contra pitiose: caracterização molecular de isolados brasileiros de Pythium insidiosum |
author |
Azevedo, Maria Isabel de |
author_facet |
Azevedo, Maria Isabel de |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Santurio, Janio Morais |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/6316012260769979 |
dc.contributor.referee1.fl_str_mv |
Leal, Daniela Bitencourt Rosa |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/3639683273462361 |
dc.contributor.referee2.fl_str_mv |
Loreto, érico Silva de |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/5475233864057995 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/7516243030317280 |
dc.contributor.author.fl_str_mv |
Azevedo, Maria Isabel de |
contributor_str_mv |
Santurio, Janio Morais Leal, Daniela Bitencourt Rosa Loreto, érico Silva de |
dc.subject.por.fl_str_mv |
Pitiose Caracterização molecular Filogenia Imunoterapia |
topic |
Pitiose Caracterização molecular Filogenia Imunoterapia Pythium insidiosum Pythiosis Phylogeny Molecular characterization Immunotherapy CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
dc.subject.eng.fl_str_mv |
Pythium insidiosum Pythiosis Phylogeny Molecular characterization Immunotherapy |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
description |
The Pythium insidiosum is an aquatic oomycete that is the causative agent of pythiosis, an important disease in humans and animals that is prevalent in tropical and subtropical areas. In Brazil it was first reported in 1974, then several cases this disease has been reported throughout the country. The failure in the antifungal therapy combined with cases unresponsive to immunotherapy existing, lead to molecular studies in order to investigate possible genetic variations among Brazilian isolates, and this may be able to contribute on the research for the improvement of immunotherapy available. Thus, this study aimed to evaluate the genetic diversity of thirty P. insidiosum isolates from different regions of Brazil, and compare all of them with isolates from Thailand, one isolate from Central America and another isolate from North America, by sequencing and the analysis of genomic DNA regions corresponding to cytochrome c oxidase (COX II) and ITS1, 5.8S rRNA and ITS2 rDNA (ITS). The analyses of nucleotide sequences of both regions were carried out, individually and in combination, using the following methods: Maximum parsimony (MP); Neighbor-joining (NJ); Maximum likelihood (ML); and Bayesian analysis (BA). Our data demonstrated all of P. insidiosum isolates as monophyletic in relation to the other Pythium species. Analises of COX II gene sequences subdivided P. insidiosum isolates into three groups, whose arrangement provides the Thai isolates as paraphyletic in relation to the Brazilian isolates. The molecular analyses performed in this study suggest an evolutionary proximity among all of American isolates, including the Brazilian, from Costa Rica and from the United States, that were grouped in a single group. COX II gene network analysis showed signs of a recent expansion of P. insidosum isolates, probably originated from the Asiatic to America continent. By analysis of COX II gene demonstrated the highest levels of genetic variability among P. insidiosum isolates studied in here, additionally, the highest levels of phylogenetic information were shown, when it was compared to the ITS region analysis. Nevertheless, both genetic markers selected for this study proved to be entirely congruent in the phylogenetic relations among Brazilian isolates of P. insidiosum, since clustered all of them into a single monophyletic group which did not shown to have genetic variability. The results indicate that the strain used in the production of the immunotherapic - Pitium-Vac ® - is representative of the Brazilian isolates of P. insidiosum. |
publishDate |
2011 |
dc.date.issued.fl_str_mv |
2011-07-15 |
dc.date.accessioned.fl_str_mv |
2012-05-08 |
dc.date.available.fl_str_mv |
2012-05-08 |
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 |
AZEVEDO, Maria Isabel de. IMMUNETHERAPY AGAINST PYTHIOSIS: MOLECULAR CHARACTERIZATION OF BRAZILIAN ISOLATES OF Pythium insidiosum. 2011. 87 f. Dissertação (Mestrado em Farmácia) - Universidade Federal de Santa Maria, Santa Maria, 2011. |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/8953 |
identifier_str_mv |
AZEVEDO, Maria Isabel de. IMMUNETHERAPY AGAINST PYTHIOSIS: MOLECULAR CHARACTERIZATION OF BRAZILIAN ISOLATES OF Pythium insidiosum. 2011. 87 f. Dissertação (Mestrado em Farmácia) - Universidade Federal de Santa Maria, Santa Maria, 2011. |
url |
http://repositorio.ufsm.br/handle/1/8953 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
201000000000 |
dc.relation.confidence.fl_str_mv |
400 500 300 500 500 |
dc.relation.authority.fl_str_mv |
06a1b29c-fde0-4a69-8097-2a9d6a9d9649 81087730-2239-428b-ad21-591665a7aecf 2dbfa8b1-3511-410e-8203-4bb0cf639ac8 ea251119-5531-432f-bf54-8c8111248727 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Farmacologia |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Farmacologia |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional Manancial 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 |
Repositório Institucional Manancial UFSM |
collection |
Repositório Institucional Manancial UFSM |
bitstream.url.fl_str_mv |
http://repositorio.ufsm.br/bitstream/1/8953/1/AZEVEDO%2c%20MARIA%20ISABEL%20DE.pdf http://repositorio.ufsm.br/bitstream/1/8953/2/AZEVEDO%2c%20MARIA%20ISABEL%20DE.pdf.txt http://repositorio.ufsm.br/bitstream/1/8953/3/AZEVEDO%2c%20MARIA%20ISABEL%20DE.pdf.jpg |
bitstream.checksum.fl_str_mv |
82fc06a0117d4629043b9d8ba576cd27 a7895a29b572186209b75fb5bb8b2da3 d3be45d5629d80e99de9ea1446cc92a2 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional Manancial UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
ouvidoria@ufsm.br |
_version_ |
1808854704187244544 |