Enterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da Canastra

Detalhes bibliográficos
Autor(a) principal: Dores, Milene Therezinha das
Data de Publicação: 2013
Tipo de documento: Tese
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/424
Resumo: The Minas artisanal cheese is, for centuries, the only link small producers with the market, so it is what gives them survival. Previous studies have shown that cheese Minas artisanal S. aureus is the species that define the minimum time needed to obtain ripening microbiological safety, since, in high counts remained for a long period during ripening. Considering that the presence of S. aureus is common in this type of cheese, the present study aimed to evaluate the presence of S. aureus enterotoxigenic endogenous starter in Minas artisanal cheeses and the Serra da Canastra. We collected 16 samples of endogenous starter and cheese in two seasons, rainy and dry. The counting and the isolation of S. aureus were performed on plate count PetrifilmTM - Rapids S. aureus. For detection of enterotoxins in cheese it was used the method in Optimal Sensitivity plates (OSP) for enterotoxinas SEA, SEB, SEC and SED and methods ELFA-VIDAS® Staph enterotoxina II for enterotoxins SEA, SEB, SEC1,2,3, SED and SEE. Strains S. aureus were tested for their ability to produce enterotoxins, using the OSP and by PCR analysis for the classical enterotoxin genes. The counts of S. aureus in cheese ripening eight days were above the limit required by Brazilian law -2.0 log CFU g-1, and ranged from 3.48 to 5.88 log CFU g-1 for rainy season and from 3.11 to 4.60 log CFU g-1 for the period of drought. Since counts of S. aureus in endogenous leaven were lower, ranging from <1 to 3.11 log CFU g-1. Only the enterotoxin SEA was detected by the OSP method, being present in 75% of samples of cheese. No toxin was detected with the method ELFA-VIDAS however false negative results were found, which may be due to the presence of possible interferences, such as substances not related to immune matrix. It was observed that 12.5% of the strains isolated produced SEA and SEC. When using the PCR assay, one isolate showed enterotoxin gene. When using the PCR assay, only two isolates enterotoxin genes (seb e seg). Although the results of the immunological and molecular methodology have been contradictory, S. aureus strains isolated from the endogenous starter and QMA showed low potential enterotoxinogênico. However, the findings in this study highlight the need for further studies to develop analytical methods that do not show interference in the results and include also the non-classical enterotoxins.
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spelling Dores, Milene Therezinha dashttp://lattes.cnpq.br/3561353982759785Furtado, Mauro Mansurhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783310Z7Pinto, Cláudia Lúcia de Oliveirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783521J6Ferreira, Célia Lúcia de Luces Forteshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4793681U9Cunha, Luciana Rodrigues dahttp://lattes.cnpq.br/3546793877060403Perrone, ítalo Tulerhttp://lattes.cnpq.br/86415123584092392015-03-26T12:24:58Z2013-05-152015-03-26T12:24:58Z2013-02-21DORES, Milene Therezinha das. Enterotoxigenicity of Staphylococcus aureus isolated of the Minas Artisanal Cheese from Canastra. 2013. 78 f. Tese (Doutorado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2013.http://locus.ufv.br/handle/123456789/424The Minas artisanal cheese is, for centuries, the only link small producers with the market, so it is what gives them survival. Previous studies have shown that cheese Minas artisanal S. aureus is the species that define the minimum time needed to obtain ripening microbiological safety, since, in high counts remained for a long period during ripening. Considering that the presence of S. aureus is common in this type of cheese, the present study aimed to evaluate the presence of S. aureus enterotoxigenic endogenous starter in Minas artisanal cheeses and the Serra da Canastra. We collected 16 samples of endogenous starter and cheese in two seasons, rainy and dry. The counting and the isolation of S. aureus were performed on plate count PetrifilmTM - Rapids S. aureus. For detection of enterotoxins in cheese it was used the method in Optimal Sensitivity plates (OSP) for enterotoxinas SEA, SEB, SEC and SED and methods ELFA-VIDAS® Staph enterotoxina II for enterotoxins SEA, SEB, SEC1,2,3, SED and SEE. Strains S. aureus were tested for their ability to produce enterotoxins, using the OSP and by PCR analysis for the classical enterotoxin genes. The counts of S. aureus in cheese ripening eight days were above the limit required by Brazilian law -2.0 log CFU g-1, and ranged from 3.48 to 5.88 log CFU g-1 for rainy season and from 3.11 to 4.60 log CFU g-1 for the period of drought. Since counts of S. aureus in endogenous leaven were lower, ranging from <1 to 3.11 log CFU g-1. Only the enterotoxin SEA was detected by the OSP method, being present in 75% of samples of cheese. No toxin was detected with the method ELFA-VIDAS however false negative results were found, which may be due to the presence of possible interferences, such as substances not related to immune matrix. It was observed that 12.5% of the strains isolated produced SEA and SEC. When using the PCR assay, one isolate showed enterotoxin gene. When using the PCR assay, only two isolates enterotoxin genes (seb e seg). Although the results of the immunological and molecular methodology have been contradictory, S. aureus strains isolated from the endogenous starter and QMA showed low potential enterotoxinogênico. However, the findings in this study highlight the need for further studies to develop analytical methods that do not show interference in the results and include also the non-classical enterotoxins.O queijo Minas artesanal é, há séculos, o único vínculo de pequenos produtores mineiros com o mercado, portanto é o que lhes garante o sustento. Estudos prévios com queijo Minas artesanais demonstraram que o S. aureus é a espécie que definiu o tempo mínimo de maturação necessário para se obter segurança microbiológica, uma vez que, permaneceu em altas contagens por um longo período durante a maturação. Considerando-se que a presença de S. aureus é comum nesse tipo de queijo, o presente estudo objetivou-se avaliar a presença de S. aureus enterotoxigênico no fermento endógeno e em queijos Minas artesanal da região da Serra da Canastra. Foram coletadas 16 amostras de fermento endógeno e de queijo em dois períodos do ano, chuvoso e seca. A contagem e o isolamento de S. aureus foram realizados em placas PetrifilmTM - Rapid S. aureus. Para a detecção de enterotoxinas em queijo utilizou-se o método de Sensibilidade Ótima em Placas (OSP) para as enterotoxinas SEA, SEB, SEC e SED e o método ELFA-VIDAS® Staph enterotoxina II, para as enterotoxinas SEA, SEB, SEC1,2,3, SED e SEE. Estirpes de S. aureus foram testadas quanto à capacidade de produzir enterotoxinas, usando o método de OSP e por meio da análise de PCR para os genes de enterotoxinas clássicas. As contagens de S. aureus nos queijos com oito dias de maturação foram superiores ao limite exigido pela legislação brasileira (2,0 log UFC.g-1) e variaram de 3,48 a 5,88 log UFC.g-1 para o período chuvoso e de 3,11 a 4,60 log UFC.g-1 para o período da seca. Já as contagens de S. aureus no fermento endógeno foram menores, variando de < 1 a 3,11 log UFC.g-1. Apenas a enterotoxina SEA foi detectada pelo método OSP, estando presente em 75% das amostras de queijo. Nenhuma toxina foi detectada com o método ELFAVIDAS, no entanto, resultados falso-negativos foram encontrados, os quais podem ser devido à presença de possíveis interferentes, como substâncias não imunológicas ligadas à matriz. Observou-se que 12,5% das estirpes isoladas produziram SEA e SEC. Ao utilizar a técnica de PCR, apenas dois isolados genes de enterotoxinas (seb e seg). Embora aos resultados entre as metodológicas imunológicas e moleculares tenha sido contraditórias, as estirpes de S.aureus isoladas do fermento endógeno e do QMA apresentaram baixo potencial enterotoxinogênico. No entanto, os resultados encontrados neste trabalho evidência a necessidade de mais estudos para o desenvolvimento de métodos analíticos que não apresentem interferência nos resultados e que incluam, também, as enterotoxinas não-clássicas.application/pdfporUniversidade Federal de ViçosaDoutorado em Ciência e Tecnologia de AlimentosUFVBRCiência de Alimentos; Tecnologia de Alimentos; Engenharia de AlimentosSerra da CanastraPCRSensibilidade ótima em placasCanastraPCROptimal Sensitivity platesCNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOSEnterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da CanastraEnterotoxigenicity of Staphylococcus aureus isolated of the Minas Artisanal Cheese from Canastrainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf1371003https://locus.ufv.br//bitstream/123456789/424/1/texto%20completo.pdff99c4dd1fe5653f2c1b9c31773df53c4MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain146942https://locus.ufv.br//bitstream/123456789/424/2/texto%20completo.pdf.txt20eb7802e6a6a23662042dd98c79394bMD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3611https://locus.ufv.br//bitstream/123456789/424/3/texto%20completo.pdf.jpg2499aa0ec4d1e4c0698fb2d0a48fc7e9MD53123456789/4242016-04-06 23:06:09.004oai:locus.ufv.br:123456789/424Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-07T02:06:09LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Enterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da Canastra
dc.title.alternative.eng.fl_str_mv Enterotoxigenicity of Staphylococcus aureus isolated of the Minas Artisanal Cheese from Canastra
title Enterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da Canastra
spellingShingle Enterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da Canastra
Dores, Milene Therezinha das
Serra da Canastra
PCR
Sensibilidade ótima em placas
Canastra
PCR
Optimal Sensitivity plates
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS
title_short Enterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da Canastra
title_full Enterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da Canastra
title_fullStr Enterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da Canastra
title_full_unstemmed Enterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da Canastra
title_sort Enterotoxigenicidade de Staphylococcus aureus isolados de queijo Minas Artesanal da Canastra
author Dores, Milene Therezinha das
author_facet Dores, Milene Therezinha das
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/3561353982759785
dc.contributor.author.fl_str_mv Dores, Milene Therezinha das
dc.contributor.advisor-co1.fl_str_mv Furtado, Mauro Mansur
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783310Z7
dc.contributor.advisor-co2.fl_str_mv Pinto, Cláudia Lúcia de Oliveira
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783521J6
dc.contributor.advisor1.fl_str_mv Ferreira, Célia Lúcia de Luces Fortes
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4793681U9
dc.contributor.referee1.fl_str_mv Cunha, Luciana Rodrigues da
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/3546793877060403
dc.contributor.referee2.fl_str_mv Perrone, ítalo Tuler
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/8641512358409239
contributor_str_mv Furtado, Mauro Mansur
Pinto, Cláudia Lúcia de Oliveira
Ferreira, Célia Lúcia de Luces Fortes
Cunha, Luciana Rodrigues da
Perrone, ítalo Tuler
dc.subject.por.fl_str_mv Serra da Canastra
PCR
Sensibilidade ótima em placas
topic Serra da Canastra
PCR
Sensibilidade ótima em placas
Canastra
PCR
Optimal Sensitivity plates
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS
dc.subject.eng.fl_str_mv Canastra
PCR
Optimal Sensitivity plates
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS
description The Minas artisanal cheese is, for centuries, the only link small producers with the market, so it is what gives them survival. Previous studies have shown that cheese Minas artisanal S. aureus is the species that define the minimum time needed to obtain ripening microbiological safety, since, in high counts remained for a long period during ripening. Considering that the presence of S. aureus is common in this type of cheese, the present study aimed to evaluate the presence of S. aureus enterotoxigenic endogenous starter in Minas artisanal cheeses and the Serra da Canastra. We collected 16 samples of endogenous starter and cheese in two seasons, rainy and dry. The counting and the isolation of S. aureus were performed on plate count PetrifilmTM - Rapids S. aureus. For detection of enterotoxins in cheese it was used the method in Optimal Sensitivity plates (OSP) for enterotoxinas SEA, SEB, SEC and SED and methods ELFA-VIDAS® Staph enterotoxina II for enterotoxins SEA, SEB, SEC1,2,3, SED and SEE. Strains S. aureus were tested for their ability to produce enterotoxins, using the OSP and by PCR analysis for the classical enterotoxin genes. The counts of S. aureus in cheese ripening eight days were above the limit required by Brazilian law -2.0 log CFU g-1, and ranged from 3.48 to 5.88 log CFU g-1 for rainy season and from 3.11 to 4.60 log CFU g-1 for the period of drought. Since counts of S. aureus in endogenous leaven were lower, ranging from <1 to 3.11 log CFU g-1. Only the enterotoxin SEA was detected by the OSP method, being present in 75% of samples of cheese. No toxin was detected with the method ELFA-VIDAS however false negative results were found, which may be due to the presence of possible interferences, such as substances not related to immune matrix. It was observed that 12.5% of the strains isolated produced SEA and SEC. When using the PCR assay, one isolate showed enterotoxin gene. When using the PCR assay, only two isolates enterotoxin genes (seb e seg). Although the results of the immunological and molecular methodology have been contradictory, S. aureus strains isolated from the endogenous starter and QMA showed low potential enterotoxinogênico. However, the findings in this study highlight the need for further studies to develop analytical methods that do not show interference in the results and include also the non-classical enterotoxins.
publishDate 2013
dc.date.available.fl_str_mv 2013-05-15
2015-03-26T12:24:58Z
dc.date.issued.fl_str_mv 2013-02-21
dc.date.accessioned.fl_str_mv 2015-03-26T12:24:58Z
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dc.identifier.citation.fl_str_mv DORES, Milene Therezinha das. Enterotoxigenicity of Staphylococcus aureus isolated of the Minas Artisanal Cheese from Canastra. 2013. 78 f. Tese (Doutorado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2013.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/424
identifier_str_mv DORES, Milene Therezinha das. Enterotoxigenicity of Staphylococcus aureus isolated of the Minas Artisanal Cheese from Canastra. 2013. 78 f. Tese (Doutorado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2013.
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