Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso
Autor(a) principal: | |
---|---|
Data de Publicação: | 2012 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFMT |
Texto Completo: | http://ri.ufmt.br/handle/1/1441 |
Resumo: | The conidiobolomycosis is a granulomatous disease associated by the fungus Conidiobolus spp. observed in humans and animals. The traditional techniques for the diagnosis of the disease are isolation of the agent associated with epidemiological aspects and the presence of typical clinical signs and pathological conditions. This work describe the development of Conidiobolus lamprauges specific PCR in order to detect the presence of fungus in clinical samples. Samples of tissue suspected of animals were collected and sent to fungal culture, histopathologic testing and PCR amplification. PCR reactions were performed using DNA extracted from fresh and paraffin embedded tissues with universal primers for fungi and amplifying fragment of the 18S region. rDNA region and specific primers were designed based on the same gene for C. lamprauges, which generated products with approximately 540 bp and 222 bp, respectively. The culture was positive in 26.6% of clinical samples. PCR tests for C. lamprauges showed DNA amplification in fresh tissue (80%) and paraffin (44.4%). Environmental samples were also collected in a property where there was an outbreak of disease, in order to carry out the study to detect possible sources of infection for sheep. During twelve months, plant and water samples were collected, their DNA extracted and analysed by to qPCR. In the analysis of environmental samples, six were positive for C. lamprauges in plant and three in water samples. The standard curve was generated resulting in a detection limit of 1 fg of DNA from C. lamprauges (3.4 x 102 molecules), ε = 0.98, R2= 0.98 and slope = -3.3. The results showed that there was no relationship between the presence of the fungus in plant material with the rainy season observed in the study. In conclusion, the PCR technique described here shows a high sensitivity and specificity for detection of C. lamprauges in clinical samples of fresh and paraffin, becoming a tool to a rapid, and the seasonality disease may not be closely related to the presence of the agent in the environment, for C. lamprauges was observed in different seasons, but were quantified at different levels during the period of study. |
id |
UFMT_85f0dbd40957ea2a1ceaa30f13af32be |
---|---|
oai_identifier_str |
oai:localhost:1/1441 |
network_acronym_str |
UFMT |
network_name_str |
Repositório Institucional da UFMT |
repository_id_str |
|
spelling |
Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato GrossoConidiobolomicosePCRqPCR18S rDNACNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAConidiobolomycosisPCRqPCR18S rDNAThe conidiobolomycosis is a granulomatous disease associated by the fungus Conidiobolus spp. observed in humans and animals. The traditional techniques for the diagnosis of the disease are isolation of the agent associated with epidemiological aspects and the presence of typical clinical signs and pathological conditions. This work describe the development of Conidiobolus lamprauges specific PCR in order to detect the presence of fungus in clinical samples. Samples of tissue suspected of animals were collected and sent to fungal culture, histopathologic testing and PCR amplification. PCR reactions were performed using DNA extracted from fresh and paraffin embedded tissues with universal primers for fungi and amplifying fragment of the 18S region. rDNA region and specific primers were designed based on the same gene for C. lamprauges, which generated products with approximately 540 bp and 222 bp, respectively. The culture was positive in 26.6% of clinical samples. PCR tests for C. lamprauges showed DNA amplification in fresh tissue (80%) and paraffin (44.4%). Environmental samples were also collected in a property where there was an outbreak of disease, in order to carry out the study to detect possible sources of infection for sheep. During twelve months, plant and water samples were collected, their DNA extracted and analysed by to qPCR. In the analysis of environmental samples, six were positive for C. lamprauges in plant and three in water samples. The standard curve was generated resulting in a detection limit of 1 fg of DNA from C. lamprauges (3.4 x 102 molecules), ε = 0.98, R2= 0.98 and slope = -3.3. The results showed that there was no relationship between the presence of the fungus in plant material with the rainy season observed in the study. In conclusion, the PCR technique described here shows a high sensitivity and specificity for detection of C. lamprauges in clinical samples of fresh and paraffin, becoming a tool to a rapid, and the seasonality disease may not be closely related to the presence of the agent in the environment, for C. lamprauges was observed in different seasons, but were quantified at different levels during the period of study.FAPEMATA conidiobolomicose é uma doença granulomatosa associada ao fungo Conidiobolus spp observada em humanos e animais. As técnicas tradicionais para o diagnóstico da doença são o isolamento do agente associado a aspectos epidemiológicos e a presença de sinais clínicos típicos e as condições patológicas. Esse trabalho descreve o desenvolvimento de PCR específico para Conidiobolus lamprauges a fim de detectar a presença do fungo em amostras clínicas. Foi realizada cultura fúngica a partir de amostras de tecidos frescos de ovinos suspeitos da doença (15), e testes histopatológicos e PCR. Amostras de tecidos parafinados (18) também foram submetidas aos testes histopatológicos e à tecnica de PCR. Foram realizadas reações de PCR a partir do DNA dos tecidos frescos e parafinados com primers universais para fungos, amplificando fragmento da região 18S rDNA e primers específicos foram desenhados com base no mesmo gene para C. lamprauges, que gerou produtos de aproximadamente 540 pb e 222 pb respectivamente. A cultura foi positiva em 26,6% das amostras clínicas. A técnica de PCR para C. lamprauges mostrou amplificação de DNA nos tecidos frescos (80%), e parafinados (44,4%). Amostras de pasto e água foram coletadas mensalmente em uma propriedade onde ocorreu surto da doença, a fim de realizar o estudo para detecção de prováveis fontes de infecção para os ovinos. Foram coletadas 12 de amostras de vegetal e água, extraido o DNA e submetidas ao qPCR. Nas analises das amostras ambientais, dos doze meses de coleta, seis mostraram-se positivos para C. lamprauges em vegetal, e três nas amostras de água. A curva-padrão foi gerada resultando em limite de detecção de 1 fg de DNA de C. lamprauges (3,4 x 102 moléculas), ε = 0.98, R2 = 0.98 e slope= -3.3. Os resultados evidenciaram que não houve relação da presença do fungo em material vegetal com o período chuvoso observado no estudo. Em conclusão, a técnica de PCR descrita aqui demonstra elevada sensibilidade e especificidade para a detecção de C. lamprauges em amostras clínicas de tecido fresco e parafinado, tornando-se uma ferramenta para um diagnóstico rápido, e que a sazonalidade da doença pode não estar estreitamente relacionada com a presença do agente no ambiente, pois C. lamprauges foi observado em diferentes épocas do ano, no entanto foram quantificados em diferentes níveis durante o periodo do estudo.Universidade Federal de Mato GrossoBrasilFaculdade de Agronomia, Medicina Veterinária e Zootecnia (FAMEVZ)UFMT CUC - CuiabáPrograma de Pós-Graduação em Ciências VeterináriasNakazato, LucianoColodel, Edson Moletahttp://lattes.cnpq.br/6150957874684255http://lattes.cnpq.br/3898850578198054Nakazato, Luciano638.389.071-91http://lattes.cnpq.br/3898850578198054Pescador, Caroline Argenta958.659.180-87http://lattes.cnpq.br/5754349416478829638.389.071-91766.442.439-91Botton, Sônia de Avila672.074.720-72http://lattes.cnpq.br/0814772095155945Silveira, Marcelo Marques da2019-09-16T09:56:03Z2012-05-032019-09-16T09:56:03Z2012-03-06info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisSILVEIRA, Marcelo Marques da. Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso. 2012. 73 f. Dissertação (Mestrado em Ciências Veterinárias) - Universidade Federal de Mato Grosso, Faculdade de Agronomia, Medicina Veterinária e Zootecnia, Cuiabá, 2012http://ri.ufmt.br/handle/1/1441porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMTinstname:Universidade Federal de Mato Grosso (UFMT)instacron:UFMT2019-09-21T07:01:54Zoai:localhost:1/1441Repositório InstitucionalPUBhttp://ri.ufmt.br/oai/requestjordanbiblio@gmail.comopendoar:2019-09-21T07:01:54Repositório Institucional da UFMT - Universidade Federal de Mato Grosso (UFMT)false |
dc.title.none.fl_str_mv |
Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso |
title |
Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso |
spellingShingle |
Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso Silveira, Marcelo Marques da Conidiobolomicose PCR qPCR 18S rDNA CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA Conidiobolomycosis PCR qPCR 18S rDNA |
title_short |
Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso |
title_full |
Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso |
title_fullStr |
Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso |
title_full_unstemmed |
Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso |
title_sort |
Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso |
author |
Silveira, Marcelo Marques da |
author_facet |
Silveira, Marcelo Marques da |
author_role |
author |
dc.contributor.none.fl_str_mv |
Nakazato, Luciano Colodel, Edson Moleta http://lattes.cnpq.br/6150957874684255 http://lattes.cnpq.br/3898850578198054 Nakazato, Luciano 638.389.071-91 http://lattes.cnpq.br/3898850578198054 Pescador, Caroline Argenta 958.659.180-87 http://lattes.cnpq.br/5754349416478829 638.389.071-91 766.442.439-91 Botton, Sônia de Avila 672.074.720-72 http://lattes.cnpq.br/0814772095155945 |
dc.contributor.author.fl_str_mv |
Silveira, Marcelo Marques da |
dc.subject.por.fl_str_mv |
Conidiobolomicose PCR qPCR 18S rDNA CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA Conidiobolomycosis PCR qPCR 18S rDNA |
topic |
Conidiobolomicose PCR qPCR 18S rDNA CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA Conidiobolomycosis PCR qPCR 18S rDNA |
description |
The conidiobolomycosis is a granulomatous disease associated by the fungus Conidiobolus spp. observed in humans and animals. The traditional techniques for the diagnosis of the disease are isolation of the agent associated with epidemiological aspects and the presence of typical clinical signs and pathological conditions. This work describe the development of Conidiobolus lamprauges specific PCR in order to detect the presence of fungus in clinical samples. Samples of tissue suspected of animals were collected and sent to fungal culture, histopathologic testing and PCR amplification. PCR reactions were performed using DNA extracted from fresh and paraffin embedded tissues with universal primers for fungi and amplifying fragment of the 18S region. rDNA region and specific primers were designed based on the same gene for C. lamprauges, which generated products with approximately 540 bp and 222 bp, respectively. The culture was positive in 26.6% of clinical samples. PCR tests for C. lamprauges showed DNA amplification in fresh tissue (80%) and paraffin (44.4%). Environmental samples were also collected in a property where there was an outbreak of disease, in order to carry out the study to detect possible sources of infection for sheep. During twelve months, plant and water samples were collected, their DNA extracted and analysed by to qPCR. In the analysis of environmental samples, six were positive for C. lamprauges in plant and three in water samples. The standard curve was generated resulting in a detection limit of 1 fg of DNA from C. lamprauges (3.4 x 102 molecules), ε = 0.98, R2= 0.98 and slope = -3.3. The results showed that there was no relationship between the presence of the fungus in plant material with the rainy season observed in the study. In conclusion, the PCR technique described here shows a high sensitivity and specificity for detection of C. lamprauges in clinical samples of fresh and paraffin, becoming a tool to a rapid, and the seasonality disease may not be closely related to the presence of the agent in the environment, for C. lamprauges was observed in different seasons, but were quantified at different levels during the period of study. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-05-03 2012-03-06 2019-09-16T09:56:03Z 2019-09-16T09:56:03Z |
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 |
SILVEIRA, Marcelo Marques da. Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso. 2012. 73 f. Dissertação (Mestrado em Ciências Veterinárias) - Universidade Federal de Mato Grosso, Faculdade de Agronomia, Medicina Veterinária e Zootecnia, Cuiabá, 2012 http://ri.ufmt.br/handle/1/1441 |
identifier_str_mv |
SILVEIRA, Marcelo Marques da. Desenvolvimento de reação em cadeia da polimerase e ecoepidemiologia de Conidiobolus lamprauges em ovinos de Alto Paraguai, Mato Grosso. 2012. 73 f. Dissertação (Mestrado em Ciências Veterinárias) - Universidade Federal de Mato Grosso, Faculdade de Agronomia, Medicina Veterinária e Zootecnia, Cuiabá, 2012 |
url |
http://ri.ufmt.br/handle/1/1441 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Mato Grosso Brasil Faculdade de Agronomia, Medicina Veterinária e Zootecnia (FAMEVZ) UFMT CUC - Cuiabá Programa de Pós-Graduação em Ciências Veterinárias |
publisher.none.fl_str_mv |
Universidade Federal de Mato Grosso Brasil Faculdade de Agronomia, Medicina Veterinária e Zootecnia (FAMEVZ) UFMT CUC - Cuiabá Programa de Pós-Graduação em Ciências Veterinárias |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFMT instname:Universidade Federal de Mato Grosso (UFMT) instacron:UFMT |
instname_str |
Universidade Federal de Mato Grosso (UFMT) |
instacron_str |
UFMT |
institution |
UFMT |
reponame_str |
Repositório Institucional da UFMT |
collection |
Repositório Institucional da UFMT |
repository.name.fl_str_mv |
Repositório Institucional da UFMT - Universidade Federal de Mato Grosso (UFMT) |
repository.mail.fl_str_mv |
jordanbiblio@gmail.com |
_version_ |
1804648497348083712 |