Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ

Detalhes bibliográficos
Ano de defesa: 2015
Autor(a) principal: Furtado, Tassia Torres lattes
Orientador(a): Lopes, Carlos Wilson Gomes lattes
Banca de defesa: Cardozo, Sergian Vianna, Almeida, Elan Cardozo Paes de
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal Rural do Rio de Janeiro
Programa de Pós-Graduação: Programa de P?s-Gradua??o em Ci?ncias Veterin?rias
Departamento: Instituto de Veterin?ria
País: Brasil
Palavras-chave em Português:
Cyniclomyces guttulatus
isolamento
caracteriza??o
c?es
Barra da Tijuca
Palavras-chave em Inglês:
Rio de Janeiro
isolation
characterization
dogs
Área do conhecimento CNPq:
Parasitologia
Link de acesso: https://tede.ufrrj.br/jspui/handle/jspui/1749
Resumo: The ascomycete yeast C.guttulatus, has long been recognized as a component of the normal microflora of rabbits and other herbivores. More recently, a possible association between this yeast and gastro-intestinal illness in dogs has been reported by researchers in Europe, EUA, Japan and Brazil. The current study combined morpho-phenotypic and molecular (PCR-RFLP and nucleotide sequencing), methods to examine C.guttulatus cultures recovered from Brazilian rabbits (15 isolates) and dogs (7 isolates), with the objectives of developing a means for the differential identification of C. guttulatus and to improve existing knowledge of the biology of this organism. Microscopic and macroscopic examination of isolated colonies in combination with zymogram analysis (carbohydrate fermentation tests), was unable to provide a definitive identification for the suspect cultures. In silico restriction fragment length polymorphism (RFLP) analysis of virtual PCR amplicons (corresponding to the D1-D2 domain of the gene encoding 26S ribosomal RNA of C.guttulatus and other yeast species known to be associated with dogs), indicated that digestion with the restriction enzymes DdeI, HaeIII and MspI would allow the differential identification of C. guttulatus. In vitro restriction analysis of PCR amplicons confirmed the predictions of the in silico analysis and demonstrated that differential identification of C.guttulatus could be a made using only two enzymes (DdeI and MspI). Sequencing of D1-D2 amplicons demonstrated the presence of substantial nucleotide variation within the cultures examined and divided them into three groups, denominated sequence types (ST?s). Two of these groups were closely related to each other. The third sequence type showed extensive nucleotide variation and may represent a novel species or subspecies within the genus Cyniclomyces
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spelling Lopes, Carlos Wilson Gomes334.954.837-72http://lattes.cnpq.br/8613149942003893McIntosh, Douglas054.046.627-19http://lattes.cnpq.br/5166697605343047Cardozo, Sergian ViannaAlmeida, Elan Cardozo Paes de108.783.337-03http://lattes.cnpq.br/1503080262726213Furtado, Tassia Torres2017-06-07T14:26:09Z2015-02-27FURTADO, Tassia Torres. Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ. 2015. 60 f. Disserta??o (Mestrado em Ci?ncias Veterin?rias) - Instituto de Veterin?ria, Universidade Federal Rural do Rio de Janeiro, Serop?dica, 2015.https://tede.ufrrj.br/jspui/handle/jspui/1749The ascomycete yeast C.guttulatus, has long been recognized as a component of the normal microflora of rabbits and other herbivores. More recently, a possible association between this yeast and gastro-intestinal illness in dogs has been reported by researchers in Europe, EUA, Japan and Brazil. The current study combined morpho-phenotypic and molecular (PCR-RFLP and nucleotide sequencing), methods to examine C.guttulatus cultures recovered from Brazilian rabbits (15 isolates) and dogs (7 isolates), with the objectives of developing a means for the differential identification of C. guttulatus and to improve existing knowledge of the biology of this organism. Microscopic and macroscopic examination of isolated colonies in combination with zymogram analysis (carbohydrate fermentation tests), was unable to provide a definitive identification for the suspect cultures. In silico restriction fragment length polymorphism (RFLP) analysis of virtual PCR amplicons (corresponding to the D1-D2 domain of the gene encoding 26S ribosomal RNA of C.guttulatus and other yeast species known to be associated with dogs), indicated that digestion with the restriction enzymes DdeI, HaeIII and MspI would allow the differential identification of C. guttulatus. In vitro restriction analysis of PCR amplicons confirmed the predictions of the in silico analysis and demonstrated that differential identification of C.guttulatus could be a made using only two enzymes (DdeI and MspI). Sequencing of D1-D2 amplicons demonstrated the presence of substantial nucleotide variation within the cultures examined and divided them into three groups, denominated sequence types (ST?s). Two of these groups were closely related to each other. The third sequence type showed extensive nucleotide variation and may represent a novel species or subspecies within the genus CyniclomycesA levedura, Cyniclomycesguttulatus ? um ascomiceto e tem sido reconhecida como um componente da microbiota natural de coelhos e outros herb?voros. Mais recentemente, uma poss?vel associa??o entre esta levedura e doen?as gastrintestinais em c?es tem sido relatada por pesquisadores na Europa, EUA, Jap?o e Brasil. O presente estudo combinou m?todos de an?lises morfol?gica e molecular (PCR-RFLP e sequenciamento de nucleot?deos), para examinar culturas de C. guttulatus recuperados de coelhos brasileiros (15 isolados) e c?es (7 isolados), com o objetivo de desenvolvimento de um m?todo para a identifica??o diferencial de C. guttulatus e melhorar o conhecimento existente sobre a biologia desse organismo. O exame microsc?pico e macrosc?pico das col?nias isoladas em combina??o com an?lise do zimograma (testes de fermenta??o de carboidratos), n?o foi capaz de fornecer uma identifica??o definitiva para as culturas suspeitas. As an?lises in silico de polimorfismo de comprimento de fragmentos de restri??o (RFLP) de produtos de PCR virtuais (correspondentes ao dom?nio D1/D2 do gene 26S RNA ribossomal de C.guttulatus e outras esp?cies de levedura conhecidas como associadas a c?es), indicou que a digest?o com a enzimas de restri??o Ddel, HaeIII e MspI permitiria a identifica??o diferencial de C. guttulatus. As an?lises das digest?es in vitro de produtos de PCR confirmaram as previs?es das an?lises in silico e demonstrou que a identifica??o diferencial de C.guttulatus poderia ser feita usando apenas duas enzimas (DdeI e MspI). O sequenciamento dos produtos de PCR do dom?nio D1/D2 demonstraram a presen?a de varia??es substanciais de nucleot?deos nas culturas analisadas e estas foram divididas em tr?s grupos, denominados de sequ?ncias tipos (ST's). Dois destes grupos estavam estreitamente relacionados uns aos outros. O terceiro tipo de sequ?ncia mostrou grande varia??o de nucleot?deos e pode representar uma nova esp?cie ou subesp?cie dentro do g?nero CyniclomycesSubmitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-06-07T14:26:09Z No. of bitstreams: 1 2015 - Tassia Torres Furtado.pdf: 14895763 bytes, checksum: 7d611b6398d83fa875abc58ca88acab3 (MD5)Made available in DSpace on 2017-06-07T14:26:09Z (GMT). No. of bitstreams: 1 2015 - Tassia Torres Furtado.pdf: 14895763 bytes, checksum: 7d611b6398d83fa875abc58ca88acab3 (MD5) Previous issue date: 2015-02-27application/pdfhttps://tede.ufrrj.br/retrieve/68358/2015%20-%20Tassia%20Torres%20Furtado%20%28parte%201%29.pdf.jpghttps://tede.ufrrj.br/retrieve/68360/2015%20-%20Tassia%20Torres%20Furtado%20%28parte%202%29.pdf.jpghttps://tede.ufrrj.br/retrieve/68362/2015%20-%20Tassia%20Torres%20Furtado%20%28parte%203%29.pdf.jpgporUniversidade Federal Rural do Rio de JaneiroPrograma de P?s-Gradua??o em Ci?ncias Veterin?riasUFRRJBrasilInstituto de Veterin?riaANVISA. Detec??o e Identifica??o dos Fungos de Import?ncia M?dica. Manual da Ag?ncia Nacional de Vigil?ncia Sanit?ria. M?dulo VII, 2004. Dispon?vel em: <www.anvisa.gov.br>. Acesso em: Abr 2013. ANVISA. Manual de Microbiologia Cl?nica para o Controle de Infec??o Relacionada ? Assist?ncia ? Sa?de. Ag?ncia Nacional de Vigil?ncia Sanit?ria, v.9, p.47 2010. Dispon?vel em: <www.anvisa.gov.br>. 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Culture, ultrastructure and physiology. 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dc.title.por.fl_str_mv Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ
dc.title.alternative.eng.fl_str_mv Isolation and characterization of Cyniclomyces guttulatus (Robin) Van Der Walt & Scott (1971) from dogs of Barra da Tijuca, Rio de Janeiro, RJ
title Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ
spellingShingle Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ
Furtado, Tassia Torres
Rio de Janeiro
isolation
characterization
dogs
Cyniclomyces guttulatus
isolamento
caracteriza??o
c?es
Barra da Tijuca
Parasitologia
title_short Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ
title_full Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ
title_fullStr Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ
title_full_unstemmed Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ
title_sort Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ
author Furtado, Tassia Torres
author_facet Furtado, Tassia Torres
author_role author
dc.contributor.advisor1.fl_str_mv Lopes, Carlos Wilson Gomes
dc.contributor.advisor1ID.fl_str_mv 334.954.837-72
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/8613149942003893
dc.contributor.advisor-co1.fl_str_mv McIntosh, Douglas
dc.contributor.advisor-co1ID.fl_str_mv 054.046.627-19
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/5166697605343047
dc.contributor.referee1.fl_str_mv Cardozo, Sergian Vianna
dc.contributor.referee2.fl_str_mv Almeida, Elan Cardozo Paes de
dc.contributor.authorID.fl_str_mv 108.783.337-03
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/1503080262726213
dc.contributor.author.fl_str_mv Furtado, Tassia Torres
contributor_str_mv Lopes, Carlos Wilson Gomes
McIntosh, Douglas
Cardozo, Sergian Vianna
Almeida, Elan Cardozo Paes de
dc.subject.eng.fl_str_mv Rio de Janeiro
isolation
characterization
dogs
topic Rio de Janeiro
isolation
characterization
dogs
Cyniclomyces guttulatus
isolamento
caracteriza??o
c?es
Barra da Tijuca
Parasitologia
dc.subject.por.fl_str_mv Cyniclomyces guttulatus
isolamento
caracteriza??o
c?es
Barra da Tijuca
dc.subject.cnpq.fl_str_mv Parasitologia
description The ascomycete yeast C.guttulatus, has long been recognized as a component of the normal microflora of rabbits and other herbivores. More recently, a possible association between this yeast and gastro-intestinal illness in dogs has been reported by researchers in Europe, EUA, Japan and Brazil. The current study combined morpho-phenotypic and molecular (PCR-RFLP and nucleotide sequencing), methods to examine C.guttulatus cultures recovered from Brazilian rabbits (15 isolates) and dogs (7 isolates), with the objectives of developing a means for the differential identification of C. guttulatus and to improve existing knowledge of the biology of this organism. Microscopic and macroscopic examination of isolated colonies in combination with zymogram analysis (carbohydrate fermentation tests), was unable to provide a definitive identification for the suspect cultures. In silico restriction fragment length polymorphism (RFLP) analysis of virtual PCR amplicons (corresponding to the D1-D2 domain of the gene encoding 26S ribosomal RNA of C.guttulatus and other yeast species known to be associated with dogs), indicated that digestion with the restriction enzymes DdeI, HaeIII and MspI would allow the differential identification of C. guttulatus. In vitro restriction analysis of PCR amplicons confirmed the predictions of the in silico analysis and demonstrated that differential identification of C.guttulatus could be a made using only two enzymes (DdeI and MspI). Sequencing of D1-D2 amplicons demonstrated the presence of substantial nucleotide variation within the cultures examined and divided them into three groups, denominated sequence types (ST?s). Two of these groups were closely related to each other. The third sequence type showed extensive nucleotide variation and may represent a novel species or subspecies within the genus Cyniclomyces
publishDate 2015
dc.date.issued.fl_str_mv 2015-02-27
dc.date.accessioned.fl_str_mv 2017-06-07T14:26:09Z
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 FURTADO, Tassia Torres. Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ. 2015. 60 f. Disserta??o (Mestrado em Ci?ncias Veterin?rias) - Instituto de Veterin?ria, Universidade Federal Rural do Rio de Janeiro, Serop?dica, 2015.
dc.identifier.uri.fl_str_mv https://tede.ufrrj.br/jspui/handle/jspui/1749
identifier_str_mv FURTADO, Tassia Torres. Isolamento e caracteriza??o de Cyniclomyces guttulatus (Robin) Van Der Walt e Scott (1971) de c?es procedentes da Barra da Tijuca, Rio de Janeiro, RJ. 2015. 60 f. Disserta??o (Mestrado em Ci?ncias Veterin?rias) - Instituto de Veterin?ria, Universidade Federal Rural do Rio de Janeiro, Serop?dica, 2015.
url https://tede.ufrrj.br/jspui/handle/jspui/1749
dc.language.iso.fl_str_mv por
language por
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