Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tede/6639 |
Resumo: | The involvement of Clostridium estertheticum as agents of deterioration of chilled beef packaged under vacuum from slaughterhouse brazilian, has been reported since 2007. Once detected, and presents the main sources of contamination in slaughter establishments, there was the need to advance the search for new knowledge about this Clostridia, however, the classical microbiological methods for their detection are considered difficult to implement because they require hard work and take a long time. PCR can be a quick and efficient detection of clostridia incriminated in the deterioration of beef chilled vacuum packaged and refrigerated environments slaughterhouses, without isolation of pure cultures. The objective of this study was to develop a test for realtime PCR for detection of Clostridium estertheticum in bovine meat and environments of slaughterhouse brazilian, as well as the molecular identification of isolates of Clostridium estertheticum by means of AFLP and RFLP techniques - PCR. The analytical results were compared with conventional PCR and confirmed by isolation. The Real-Time PCR assay for detection of Clostridium estertheticum in bovine meat and environments of slaughterhouse brazilian, was efficient. Evidence for the presence and distribution of C. estertheticum the sources studied and the technique of RT-PCR provided the highest percentage of positive samples from slaughterhouses that refrigerators Conventional PCR, and this is confirmed by isolation, which has become an important alternative for the detection of C. estertheticum in bovine meat and environments of slaughterhouse without isolation. The isolates used showed general similarity of 42.4%, suggesting high genetic diversity. In addition, the isolates obtained from samples of ambient and refrigerated meat tufted not show significant similarity, greater than 80%, used with any typical strain. The similarity found between the isolates may be indicative of possible similarity between those from meats and puffed from between those environments and equipment. The study revealed that additional steps necessary x for the characterization of Brazilian isolates, especially sequencing of other regions of the DNA so as to determine their taxonomic position. |
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Mesquita, Albenones Jose dehttp://lattes.cnpq.br/3180029815183858Mesquita, Albenones Jose deAndré, Maria Claudia Dantas Porfírio BorgesBataus, Luiz Artur MendesMazzoni, Rolando AlfredoSoares, Renata de Bastos Ascençohttp://lattes.cnpq.br/7984848783062844Marra, Kelly Nobre2016-12-26T14:21:56Z2012-05-09MARRA, K. N. Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros. 2012. 78 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás, Goiânia, 2012.http://repositorio.bc.ufg.br/tede/handle/tede/6639The involvement of Clostridium estertheticum as agents of deterioration of chilled beef packaged under vacuum from slaughterhouse brazilian, has been reported since 2007. Once detected, and presents the main sources of contamination in slaughter establishments, there was the need to advance the search for new knowledge about this Clostridia, however, the classical microbiological methods for their detection are considered difficult to implement because they require hard work and take a long time. PCR can be a quick and efficient detection of clostridia incriminated in the deterioration of beef chilled vacuum packaged and refrigerated environments slaughterhouses, without isolation of pure cultures. The objective of this study was to develop a test for realtime PCR for detection of Clostridium estertheticum in bovine meat and environments of slaughterhouse brazilian, as well as the molecular identification of isolates of Clostridium estertheticum by means of AFLP and RFLP techniques - PCR. The analytical results were compared with conventional PCR and confirmed by isolation. The Real-Time PCR assay for detection of Clostridium estertheticum in bovine meat and environments of slaughterhouse brazilian, was efficient. Evidence for the presence and distribution of C. estertheticum the sources studied and the technique of RT-PCR provided the highest percentage of positive samples from slaughterhouses that refrigerators Conventional PCR, and this is confirmed by isolation, which has become an important alternative for the detection of C. estertheticum in bovine meat and environments of slaughterhouse without isolation. The isolates used showed general similarity of 42.4%, suggesting high genetic diversity. In addition, the isolates obtained from samples of ambient and refrigerated meat tufted not show significant similarity, greater than 80%, used with any typical strain. The similarity found between the isolates may be indicative of possible similarity between those from meats and puffed from between those environments and equipment. The study revealed that additional steps necessary x for the characterization of Brazilian isolates, especially sequencing of other regions of the DNA so as to determine their taxonomic position.O envolvimento de Clostridium estertheticum como agente de deterioração de carnes bovinas refrigeradas embaladas a vácuo provenientes de ambientes de frigoríficos no Brasil, vem sendo relatado desde 2007. Uma vez detectados, bem como apontadas as principais fontes de contaminação nos estabelecimentos de abate, surgiu à necessidade de avançar na busca de novos conhecimentos sobre esses clostrídios. No entanto, os métodos microbiológicos clássicos para sua detecção são considerados de difícil execução, pois exigem muito trabalho e demandam muito tempo. A PCR pode ser uma alternativa rápida e eficiente para detecção de clostrídios incriminados na deterioração de carnes bovinas refrigeradas embaladas a vácuo e ambientes de matadouros-frigoríficos, sem a necessidade de isolamento em culturas puras. Objetivou-se com o presente estudo desenvolver um ensaio de PCR em Tempo Real para detecção de Clostridium estertheticum em carnes bovinas e superfícies diversas de matadouros-frigoríficos brasileiros, assim como a identificação molecular de isolados de Clostridium estertheticum, por meio das técnicas RFLP e AFLP – PCR. Os resultados analíticos foram comparados com PCR Convencional e confirmados pelo isolamento. O ensaio PCR em Tempo Real desenvolvido para detecção de Clostridium estertheticum em carnes bovinas e superfícies diversas de matadouros-frigoríficos brasileiros, mostrou-se eficiente. Evidencia-se a presença e disseminação de C. estertheticum nas fontes estudadas e a técnica de PCR em Tempo Real proporcionou maior percentual de positividade para amostras de matadouros- frigoríficos que a técnica de PCR Convencional, sendo este confirmado pelo isolamento, o que demonstra ser uma alternativa importante na detecção de C. estertheticum de carnes bovinas e superfícies diversas de matadouros-frigoríficos sem a necessidade de isolamento. Os isolados utilizados revelaram similaridade geral de 42,4%, sugerindo alta diversidade genética. Além viii disso, os isolados obtidos a partir de amostras de ambientes frigoríficos e carnes tufadas não apresentaram similaridade significativa, maior que 80%, com nenhuma cepa padrão utilizada. A similaridade encontrada entre os isolados pode ser indicativa de possível semelhança entre aqueles provenientes de carnes tufadas e entre aqueles provenientes de ambientes e equipamentos. O estudo mostrou serem necessárias etapas complementares de caracterização dos isolados brasileiros, principalmente sequenciamento de outras regiões do DNA para assim determinar sua posição taxonômica.Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2016-12-21T16:24:41Z No. of bitstreams: 2 Tese - Kelly Nobre Marra - 2012.pdf: 2565911 bytes, checksum: f0057800bcb5bba97865ae5ccc3e4507 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-12-26T14:21:56Z (GMT) No. of bitstreams: 2 Tese - Kelly Nobre Marra - 2012.pdf: 2565911 bytes, checksum: f0057800bcb5bba97865ae5ccc3e4507 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2016-12-26T14:21:56Z (GMT). No. of bitstreams: 2 Tese - Kelly Nobre Marra - 2012.pdf: 2565911 bytes, checksum: f0057800bcb5bba97865ae5ccc3e4507 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2012-05-09Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESapplication/pdfporUniversidade Federal de GoiásPrograma de Pós-graduação em Ciência Animal (EVZ)UFGBrasilEscola de Veterinária e Zootecnia - EVZ (RG)http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessCarneDeterioraçãoPCR em Tempo RealTufamentoBlown packDeteriorationMeatReal-Time PCRCIENCIA E TECNOLOGIA DE ALIMENTOS::TECNOLOGIA DE ALIMENTOSDetecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileirosDetection by PCR realtime and molecular identification of clostridium estertheticum in bovine meat and environments of slaughterhouse brazilianinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis4581960685150189167600600600600-6217552114249094582-76365128114794953382075167498588264571reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; 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dc.title.por.fl_str_mv |
Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros |
dc.title.alternative.eng.fl_str_mv |
Detection by PCR realtime and molecular identification of clostridium estertheticum in bovine meat and environments of slaughterhouse brazilian |
title |
Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros |
spellingShingle |
Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros Marra, Kelly Nobre Carne Deterioração PCR em Tempo Real Tufamento Blown pack Deterioration Meat Real-Time PCR CIENCIA E TECNOLOGIA DE ALIMENTOS::TECNOLOGIA DE ALIMENTOS |
title_short |
Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros |
title_full |
Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros |
title_fullStr |
Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros |
title_full_unstemmed |
Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros |
title_sort |
Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros |
author |
Marra, Kelly Nobre |
author_facet |
Marra, Kelly Nobre |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Mesquita, Albenones Jose de |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/3180029815183858 |
dc.contributor.referee1.fl_str_mv |
Mesquita, Albenones Jose de |
dc.contributor.referee2.fl_str_mv |
André, Maria Claudia Dantas Porfírio Borges |
dc.contributor.referee3.fl_str_mv |
Bataus, Luiz Artur Mendes |
dc.contributor.referee4.fl_str_mv |
Mazzoni, Rolando Alfredo |
dc.contributor.referee5.fl_str_mv |
Soares, Renata de Bastos Ascenço |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/7984848783062844 |
dc.contributor.author.fl_str_mv |
Marra, Kelly Nobre |
contributor_str_mv |
Mesquita, Albenones Jose de Mesquita, Albenones Jose de André, Maria Claudia Dantas Porfírio Borges Bataus, Luiz Artur Mendes Mazzoni, Rolando Alfredo Soares, Renata de Bastos Ascenço |
dc.subject.por.fl_str_mv |
Carne Deterioração PCR em Tempo Real Tufamento |
topic |
Carne Deterioração PCR em Tempo Real Tufamento Blown pack Deterioration Meat Real-Time PCR CIENCIA E TECNOLOGIA DE ALIMENTOS::TECNOLOGIA DE ALIMENTOS |
dc.subject.eng.fl_str_mv |
Blown pack Deterioration Meat Real-Time PCR |
dc.subject.cnpq.fl_str_mv |
CIENCIA E TECNOLOGIA DE ALIMENTOS::TECNOLOGIA DE ALIMENTOS |
description |
The involvement of Clostridium estertheticum as agents of deterioration of chilled beef packaged under vacuum from slaughterhouse brazilian, has been reported since 2007. Once detected, and presents the main sources of contamination in slaughter establishments, there was the need to advance the search for new knowledge about this Clostridia, however, the classical microbiological methods for their detection are considered difficult to implement because they require hard work and take a long time. PCR can be a quick and efficient detection of clostridia incriminated in the deterioration of beef chilled vacuum packaged and refrigerated environments slaughterhouses, without isolation of pure cultures. The objective of this study was to develop a test for realtime PCR for detection of Clostridium estertheticum in bovine meat and environments of slaughterhouse brazilian, as well as the molecular identification of isolates of Clostridium estertheticum by means of AFLP and RFLP techniques - PCR. The analytical results were compared with conventional PCR and confirmed by isolation. The Real-Time PCR assay for detection of Clostridium estertheticum in bovine meat and environments of slaughterhouse brazilian, was efficient. Evidence for the presence and distribution of C. estertheticum the sources studied and the technique of RT-PCR provided the highest percentage of positive samples from slaughterhouses that refrigerators Conventional PCR, and this is confirmed by isolation, which has become an important alternative for the detection of C. estertheticum in bovine meat and environments of slaughterhouse without isolation. The isolates used showed general similarity of 42.4%, suggesting high genetic diversity. In addition, the isolates obtained from samples of ambient and refrigerated meat tufted not show significant similarity, greater than 80%, used with any typical strain. The similarity found between the isolates may be indicative of possible similarity between those from meats and puffed from between those environments and equipment. The study revealed that additional steps necessary x for the characterization of Brazilian isolates, especially sequencing of other regions of the DNA so as to determine their taxonomic position. |
publishDate |
2012 |
dc.date.issued.fl_str_mv |
2012-05-09 |
dc.date.accessioned.fl_str_mv |
2016-12-26T14:21:56Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
MARRA, K. N. Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros. 2012. 78 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás, Goiânia, 2012. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/6639 |
identifier_str_mv |
MARRA, K. N. Detecção por PCR em tempo real e identificação molecular de clostridium estertheticum proveniente de carne bovina e ambientes de matadouros frigoríficos brasileiros. 2012. 78 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás, Goiânia, 2012. |
url |
http://repositorio.bc.ufg.br/tede/handle/tede/6639 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.program.fl_str_mv |
4581960685150189167 |
dc.relation.confidence.fl_str_mv |
600 600 600 600 |
dc.relation.department.fl_str_mv |
-6217552114249094582 |
dc.relation.cnpq.fl_str_mv |
-7636512811479495338 |
dc.relation.sponsorship.fl_str_mv |
2075167498588264571 |
dc.rights.driver.fl_str_mv |
http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.publisher.program.fl_str_mv |
Programa de Pós-graduação em Ciência Animal (EVZ) |
dc.publisher.initials.fl_str_mv |
UFG |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Escola de Veterinária e Zootecnia - EVZ (RG) |
publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFG instname:Universidade Federal de Goiás (UFG) instacron:UFG |
instname_str |
Universidade Federal de Goiás (UFG) |
instacron_str |
UFG |
institution |
UFG |
reponame_str |
Repositório Institucional da UFG |
collection |
Repositório Institucional da UFG |
bitstream.url.fl_str_mv |
http://repositorio.bc.ufg.br/tede/bitstreams/04423c7a-fe32-4101-8cfe-1aeabf980cea/download http://repositorio.bc.ufg.br/tede/bitstreams/47da0ebe-5051-4e6c-b2ff-0d6b0b499f52/download http://repositorio.bc.ufg.br/tede/bitstreams/64c4e37b-336a-4413-9d63-fbe9b79e4b7a/download http://repositorio.bc.ufg.br/tede/bitstreams/fbc42b65-f874-4c7c-8b52-c3e7e55a86e5/download http://repositorio.bc.ufg.br/tede/bitstreams/006ef212-4d67-4272-b08e-a5497ce90b56/download |
bitstream.checksum.fl_str_mv |
bd3efa91386c1718a7f26a329fdcb468 4afdbb8c545fd630ea7db775da747b2f d41d8cd98f00b204e9800998ecf8427e d41d8cd98f00b204e9800998ecf8427e f0057800bcb5bba97865ae5ccc3e4507 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional da UFG - Universidade Federal de Goiás (UFG) |
repository.mail.fl_str_mv |
tasesdissertacoes.bc@ufg.br |
_version_ |
1793965521523179520 |