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dc.contributor.authorSoleiro, Carla Alves
dc.date.accessioned2023-12-21T18:45:46Z-
dc.date.available2023-12-21T18:45:46Z-
dc.date.issued2012-12-10
dc.identifier.citationSOLEIRO, Carla Alves. Caracterização de Aspergillus fumigatus isolados de diferentes origens quanto ao perfil enzimático, genético e a produção de gliotoxina. 2012. 52 f. Tese (Doutorado em Ciência, Tecnologia e Inovação em Agropecuária) - Pró-Reitoria de Pesquisa e Pós-Graduação, Universidade Federal Rural do Rio de Janeiro, Seropédica, 2012.por
dc.identifier.urihttps://rima.ufrrj.br/jspui/handle/20.500.14407/9820-
dc.description.abstractAlgumas espécies do gênero Aspergillus podem causar doenças em humanos e animais como agente etiológico ou pela produção de metabólitos secudários, as micotoxinas. A contaminação por essas toxinas é um problema na produção de alimentos e no armazenamento destes. A gliotoxina possui vários papéis imunossupressivos além de poder estar envolvida no início do proceso infeccioso causado por A. fumigatus. Na natureza, parte da atividade enzimática necessária para o aproveitamento da matéria orgânica é realizada por fungos filamentosos, e elas têm grande importância fisiológica. Aspergillus fumigatus é o agente etiológico mais implicado na Aspergilose Invasiva (AI) humana, no entanto a identificação dessa espécie tem sido baseada nas características morfológicas, muitas vezes de forma errônea. Ultimamente, têm sido desenvolvidos métodos de identificação de fungos toxígenos baseados em técnicas moleculares. Os objetivos desse estudo foram: avaliar a capacidade de cepas de A. fumigatus isoladas de diferentes origens produzirem gliotoxina; estabelecer as diferenças enzimáticas entre elas, e identificar geneticamente essas cepas, além de estabelecer possíveis influências que as diferentes origens pudessem exercer sobre essas cepas. Foram utilizadas 53 cepas identificadas morfologicamente por A. fumigatus pertencentes ao Núcleo de Pesquisas Micológicas e Mitoxicológicas, isoladas de ração para consumo animal, cereais, silagens, amostras clínicas humana e animal. Para detecção e quantificação da capacidade de produção de gliotoxina foi utilizada a Cromatografia Líquida de Alta Eficiência, as análises enzimáticas foram qualitativas e a caracterização genética foi realizada através da técnica Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP). Foi utilizada a Análise em Componentes Principais expressa em gráficos Biplot. Foi possível detectar a produção de gliotoxina em todas as cepas isoladas de silagem de milho, amostras clínicas humana e animal. As cepas isoladas de amostras clínicas humanas e de silagem de milho foram as que mais produziram gliotoxina. A presença de açúcar redutor na hidrólise do amido sofreu influência do tempo, já que este foi constatado aos 14 dias em 86 % das cepas de silagem de sorgo, 100 % das amostras clínica humana e 75 % das amostras clínica animal. Houve uma diferença nos resultados quanto a produção de celulase, já que no papel filtro apenas uma cepa foi negativa, enquanto dez cepas a produziram no agar carboximetilcelulose. Quanto a produção de caseinase, 23 % produziram essa enzima. As cepas isoladas de origem clínica (animal e humana) foram as que mais apresentaram capacidade para hidrolisar a gelatina. As cepas de origem clínica animal (isolada do úbere da vaca com mastite bovina) foram melhores caracterizadas pela variável produção de caseinase, enquanto as cepas de amostras clínica humana e as de silagem de milho foram melhores caracterizadas pela variável produção de gliotoxina. Todas as cepas morfologicamente identificadas como A. fumigatus produziram um padrão de bandas correspondente a identificação da espécie A. fumigatus strictu sensu pelo corte in silico e pela técnica de PCR-RFLP.por
dc.description.sponsorshipCAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorpor
dc.formatapplication/pdf*
dc.languageporpor
dc.publisherUniversidade Federal Rural do Rio de Janeiropor
dc.rightsAcesso Abertopor
dc.subjectAspergillus fumigatuspor
dc.subjectMicotoxinapor
dc.subjectAnálises enzimática e molecularpor
dc.subjectAspergillus fumigateseng
dc.subjectMycotoxineng
dc.subjectMolecular and enzimatic analysiseng
dc.titleCaracterização de Aspergillus fumigatus isolados de diferentes origens quanto ao perfil enzimático, genético e a produção de gliotoxinapor
dc.title.alternativeCharacterization of Aspergillus fumigatus isolates from different sources regarding enzymatic, genetic and production of gliotoxin profileseng
dc.typeTesepor
dc.description.abstractOtherThe genus Aspergillus is a filamentous fungus found in all parts of the world, some species can cause illness in humans and animals. Contamination with mycotoxins is a problem in food production and storage. The gliotoxin has several immunosuppressive roles. Recently, it was found in conidia of A. fumigatus, which may indicate their involvement at the beginning of infectious disease. In nature, part of the enzymatic activity required for the utilization of organic material is performed by filamentous fungi, and they have great physiological importance. Aspergillus fumigatus is the etiological agent most involved in human Invasive Aspergillosis (IA), however the identification of this specie has been based on morphological characteristics. That identification can be a problem, so the development of rapid and sensitive methods for the correct identificacion of the fungi, such as the molecular techniques is relevant. The objectives of this study were: to evaluate the ability of A. fumigatus strains isolates from different origins to produce gliotoxin, and to establish the physiological and genetic differences of these strains and how they are influenced by different sources. A total of 53 isolates identified morphologically by A. fumigatius, belonging to the Nucleus of Mycological and Mycotoxicological Researchs, were isolated from: feed for animal consumption, cereal grains, corn and sorghum silages, clinicals human and animals. For detection and quantification the production of gliotoxin HPLC was used. Genetic analysis was performed by PCR-RFLP. Principal Component Analysis (PCA) was used to obtain a smaller number of variables able to express the variability of the data. It was possible to detect the production of gliotoxin in all strains of corn silage, clinical human and animal. Among the seven strains of human clinical, six produced more than 20 μg/g of the toxin. The strains isolated from corn silage and human clinical were the most produced gliotoxin. The presence of reducing sugars was influenced by time, since it was found at 14 days in 86% of strains of sorghum silage, 100% of clinical human and 75% of bovine mastitis. Only one strain of animal feed did not present conidia in the filter paper, so it was negative for cellulose hydrolysis. However, in the CMC Agar, ten strains did not hydrolyzed cellulose and 70% of these were isolated from animal feed. Almost all the strains (98%) isolated of the studied produced lipase. As for the casein hydrolysis 23% strains were positive, and those isolated from bovine mastitis were the most positive. The strains of clinical origin (animal and human) were the ones that showed highest ability to hydrolyze gelatin. According to the PCA, the clinical animal strains were better characterized by variable hydrolysis of casein, and the human clinical strains and corn silage were best characterized by the variable production of gliotoxin. All strains morphologically identified as A. fumigatus have produced a pattern of bands corresponding to the identification of species A. fumigatus strictu sensu by in silico cut and by PCR-RFLP technique.eng
dc.contributor.advisor1Rosa, Carlos Alberto da Rocha
dc.contributor.advisor1ID362.637.537-49por
dc.contributor.advisor1Latteshttp://lattes.cnpq.br/2073046127303600por
dc.contributor.advisor-co1Cavaglieri, Lilia Reneé
dc.contributor.referee1Dalcero, Ana
dc.contributor.referee2Keller, Kelly Moura
dc.contributor.referee3McIntosh, Douglas
dc.contributor.referee4Coelho, Irene da Silva
dc.contributor.referee5Rubinstein, Héctor Ramón
dc.creator.ID095.455.507-40por
dc.creator.Latteshttp://lattes.cnpq.br/0310733047446961por
dc.publisher.countryBrasilpor
dc.publisher.departmentPró-Reitoria de Pesquisa e Pós-Graduaçãopor
dc.publisher.initialsUFRRJpor
dc.publisher.programPrograma de Pós-Graduação em Ciência, Tecnologia e Inovação em Agropecuáriapor
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