Please use this identifier to cite or link to this item: https://rima.ufrrj.br/jspui/handle/20.500.14407/9582
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dc.contributor.authorFurtado, Tassia Torres
dc.date.accessioned2023-12-21T18:41:48Z-
dc.date.available2023-12-21T18:41:48Z-
dc.date.issued2020-04-29
dc.identifier.citationFURTADO, Tassia Torres. Análise genética e desenvolvimento de um método de diagnóstico, baseado em PCR, para detecção de Cyniclomyces guttulatus em fezes de cães. 2020. 115 f. Tese (Doutorado em Ciências Veterinárias) - Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, 2020.por
dc.identifier.urihttps://rima.ufrrj.br/jspui/handle/20.500.14407/9582-
dc.description.abstractO significado das infecções fúngicas dos animais tem sido ignorado por muitos veterinários. No entanto, problemas recentes causados por doenças fúngicas de destaque, por exemplo, a esporotricose de felinos e humanos no Brasil e a criptococose zoonótica em todo o mundo, demonstraram a necessidade de reverter essa tendência negativa há muito estabelecida. A levedura ascomicética Cyniclomyces guttulatus é a única espécie atualmente incluída no gênero Cyniclomyces. Tradicionalmente, era vista como um componente comensal da microbiota intestinal de coelhos e outros herbívoros. O aumento do interesse em C. guttulatus se desenvolveu com base em relatos da Europa, Ásia, América do Norte e Brasil, sugerindo que C. guttulatus representava um patógeno emergente e / ou oportunista de cães, onde sua presença em números elevados estava ligada a sinais clínicos, incluindo diarreia, gastroenterite e colangiohepatite. No entanto, pesquisas recentes conduzidas na Holanda e nos Estados Unidos argumentaram que a presença dessa levedura em cães que sofrem de doença gastrintestinal é provavelmente circunstancial e não tem relevância clínica. O diagnóstico de C. guttulatus em fezes e / ou em material clínico é realizado principalmente usando análises microscópicas e microbiológicas baseadas em cultura. Em uma minoria de estudos, a identificação definitiva das colônias da levedura foi feita utilizando a amplificação por reação em cadeia da polimerase (PCR) e sequenciamento de um fragmento do gene 26S rDNA (codifica a subunidade maior do RNA ribossomal). O presente estudo relata o desenvolvimento de métodos moleculares inovadores para detecção e caracterização de culturas de leveduras e para a detecção e caracterização independente de cultura de C. guttulatus diretamente de fezes de cães. A disponibilidade dos ensaios servirá para reduzir as dúvidas sobre a validade dos diagnósticos morfológicos e pode reduzir o tempo para confirmação do diagnóstico molecular para até 48 horas, ao invés do prazo atual de 7 a 10 dias. A genotipagem de uma coleção de isolados de porquinho-da-índia (n = 1), coelhos (n = 20) e cães (n = 8) utilizou amplificação por PCR e sequenciamento de fragmentos das sequências que codificam as subunidades maior (26S) e menor (18S) do RNA ribossomal, as subunidades RPB1 e RPB2 da RNA polimerase II, a citocromo oxidase II e as regiões espaçadoras internas transcritas (ITS). Alinhamento de sequências e avaliação da similaridade entre os nucleotídeos para a maioria das sequências serviram para apoiar dados de estudos anteriores que sugeriram a possível existência de duas espécies de Cyniclomyces. Curiosamente, o uso da região ITS indicou que pode haver pelo menos três espécies de Cyniclomyces. Conclui-se que a identificação das novas espécies de Cyniclomyces questiona grande parte dos dados produzidos até o momento em relação à patogenicidade desse organismo. Prevê-se que os resultados deste trabalho sirvam para estimular uma expansão do estudo do gênero Cyniclomyces e, como tal, contribuam significativamente para a compreensão do seu papel na saúde dos cães.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.subjectlevedurapor
dc.subjectbiologia molecularpor
dc.subjectdoença gastrintestinalpor
dc.subjectcaninopor
dc.subjectyeasteng
dc.subjectmolecular biologyeng
dc.subjectgastrointestinal illnesseng
dc.subjectcanineeng
dc.titleAnálise genética e desenvolvimento de um método de diagnóstico, baseado em PCR, para detecção de Cyniclomyces guttulatus em fezes de cãespor
dc.title.alternativeGenetic analysis and development of a PCR based diagnostic method for detection of Cyniclomyces guttulatus in dogs feceseng
dc.typeTesepor
dc.description.abstractOtherMany veterinary scientists have largely overlooked the significance of fungal infections in animals. However, recent high-profile fungal disease problems e.g. sporotrichosis of felines and humans in Brazil and zoonotic cryptococcosis worldwide, have demonstrated the need to revert this long-established negative trend. The ascomycete yeast Cyniclomyces guttulatusis is the only species currently included in the genus Cyniclomyces. Traditionally, it was viewed as a commensal component of the gut microbiota of rabbits and other herbivores. Increased interest in C. guttulatus developed based on reports from Europe, Asia, North America and Brazil suggesting that C. guttulatus represented an emerging and/or opportunistic pathogen in dogs, where its presence, in large numbers, was linked to clinical signs including diarrhea, gastroenteritis and cholangiohepatitis. However, recent investigations conducted in the Netherlands and the United States have argued that the presence of this yeast in dogs suffering gastrointestinal disease is most likely circumstantial and is not of clinical relevance. Diagnosis of C. guttulatus in feces and/or in clinical material is conducted primarily using microscopic and microbiological culture-based analyses. In a minority of studies, definitive identification of yeast colonies was made using polymerase chain reaction (PCR) amplification and sequencing of a fragment of the universal yeast gene 26S (encodes for the large subunit of ribosomal RNA). The current study reports the development of innovative, molecular methods for the detection and characterization of yeast cultures and for the culture-independent detection and characterization of C. guttulatus directly from canine feces. The availability of the analyses will serve to reduce doubt over the validity of morphological diagnostics and can reduce the time for a confirmed molecular diagnosis to 48 hours, rather than the actual timescale of 7-10 days. Genotyping of a collection of isolates from guinea pigs (n=1), rabbits (n= 20) and dogs (n=8), used PCR-based amplification and sequencing of fragments of the sequences encoding the large (26S) and small (18S) subunits of ribosomal rDNA, the subunits RPB1 and RPB2 of the gene encoding RNA polymerase II, Cytochrome oxidase II and the internal transcribed spacer (ITS) regions of the nuclear ribosomal RNA gene cluster. Sequence alignments and evaluations of nucleotide similarity for most sequences, served to support data from earlier studies, which had suggested the possible existence of two species of Cyniclomyces. Intriguingly, the use of the ITS region indicated that there may exist at least three species of Cyniclomyces. The identification of the new species of Cyniclomyces calls into question much of the data produced until now in relation to the pathogenicity of this organism. It is envisaged that the findings of this research will serve to stimulate an expansion in the study of the genus Cyniclomyces, and as such will make meaningful contributions to our comprehension of its role in canine health.eng
dc.contributor.advisor1McIntosh, Douglas
dc.contributor.advisor1ID054.046.627-19por
dc.contributor.advisor1Latteshttp://lattes.cnpq.br/5166697605343047por
dc.contributor.advisor-co1Lopes, Carlos Wilson Gomes
dc.contributor.advisor-co1ID334.954.837-72por
dc.contributor.advisor-co1Latteshttp://lattes.cnpq.br/8613149942003893por
dc.contributor.referee1Mclntosh, Douglas
dc.contributor.referee2Baroni, Francisco de Assis
dc.contributor.referee3Santos, Leandro Azevedo
dc.contributor.referee4Santos, Fábio Brito dos
dc.contributor.referee5Schwab, Stefan
dc.creator.ID108.783.337-03por
dc.creator.IDhttps://orcid.org/0000-0003-4439-0218por
dc.creator.Latteshttp://lattes.cnpq.br/1503080262726213por
dc.publisher.countryBrasilpor
dc.publisher.departmentInstituto de Veterináriapor
dc.publisher.initialsUFRRJpor
dc.publisher.programPrograma de Pós-Graduação em Ciências Veterináriaspor
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