Expressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianos

Detalhes bibliográficos
Autor(a) principal: Klein, Raphael Contelli
Data de Publicação: 2010
Tipo de documento: Dissertação
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/2426
Resumo: Staphylococcus aureus is one of the main micro-organisms causing bovine mastitis, a disease that causes the greatest losses in dairy farming worldwide. This pathogen has several virulence factors that contribute to the great genetic diversity observed among isolates and assist in the establishment of infections. In recent years, several studies have shown that the use of antibiotics in subinhibitory concentrations modulates gene expression, influencing the virulence of bacterial pathogens. This study aimed to investigate the effect of subinhibitory concentrations of ampicillin, gentamicin, oxacillin and tylosin, antibiotics used in veterinary formulations for the treatment of mastitis, in the expression of genes from two strains of S. aureus of bovine origin. Initially, we conducted a preliminary investigation of the presence of genes encoding some virulence factors in 85 bacteria isolated from animals with bovine mastitis outbreak. Genes clfB and sdrCDE were the most prevalent, detected in 83.5% and 75.3% of the isolates, respectively. The genetic diversity of isolates, assessed by multiplex PCR, was also high and allowed the discrimination of more than 60 groups. These results guided the choice of S. aureus 4006 and 4125, with genetic similarity of 80% for the later trials. Total RNA from two cultures grown in values equivalent to 0.5X, 0.25X, 0.125X and the minimum inhibitory concentration defined for the four antibiotics was extracted and used in the analysis of real time RT-PCR. The expression of genes spa, clfB, sdrC, fnBP, icaD, icaR, murf and sarA was normalized to the gyrB gene. All antibiotics tested caused alterations in the expression of genes evaluated. The same antibiotics used in different doses, as well as different antibiotics at the same dose, differentially modulate the expression of genes. Only for 4006, the SarA regulatory protein, which regulates the transcription of several virulence factors, was expressed in many different treatments. As for 4125 isolate, the icaD regulator gene was positively influenced when different conditions were tested. In short, the data confirm that genes of S. aureus are expressed differently in response to concentrations of antibiotics below the considered inhibitory concentration and that the variance between strains of S. aureus makes difficult a pattern of expression is extended to all species.
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spelling Klein, Raphael Contellihttp://lattes.cnpq.br/7185650565356570Bazzolli, Denise Mara Soareshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4761710D6Fietto, Luciano Gomeshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763824H8Ribon, Andréa de Oliveira Barroshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026E6Zerbini, Poliane Alfenashttp://lattes.cnpq.br/8632328159533071Fietto, Juliana Lopes Rangelhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790238D02015-03-26T13:07:31Z2011-10-212015-03-26T13:07:31Z2010-02-23KLEIN, Raphael Contelli. Gene expression of Staphylococcus aureus isolated from bovine mastitis in response to subinhibitory concentrations of antibiotics. 2010. 82 f. Dissertação (Mestrado em Bioquímica e Biologia molecular de plantas; Bioquímica e Biologia molecular animal) - Universidade Federal de Viçosa, Viçosa, 2010.http://locus.ufv.br/handle/123456789/2426Staphylococcus aureus is one of the main micro-organisms causing bovine mastitis, a disease that causes the greatest losses in dairy farming worldwide. This pathogen has several virulence factors that contribute to the great genetic diversity observed among isolates and assist in the establishment of infections. In recent years, several studies have shown that the use of antibiotics in subinhibitory concentrations modulates gene expression, influencing the virulence of bacterial pathogens. This study aimed to investigate the effect of subinhibitory concentrations of ampicillin, gentamicin, oxacillin and tylosin, antibiotics used in veterinary formulations for the treatment of mastitis, in the expression of genes from two strains of S. aureus of bovine origin. Initially, we conducted a preliminary investigation of the presence of genes encoding some virulence factors in 85 bacteria isolated from animals with bovine mastitis outbreak. Genes clfB and sdrCDE were the most prevalent, detected in 83.5% and 75.3% of the isolates, respectively. The genetic diversity of isolates, assessed by multiplex PCR, was also high and allowed the discrimination of more than 60 groups. These results guided the choice of S. aureus 4006 and 4125, with genetic similarity of 80% for the later trials. Total RNA from two cultures grown in values equivalent to 0.5X, 0.25X, 0.125X and the minimum inhibitory concentration defined for the four antibiotics was extracted and used in the analysis of real time RT-PCR. The expression of genes spa, clfB, sdrC, fnBP, icaD, icaR, murf and sarA was normalized to the gyrB gene. All antibiotics tested caused alterations in the expression of genes evaluated. The same antibiotics used in different doses, as well as different antibiotics at the same dose, differentially modulate the expression of genes. Only for 4006, the SarA regulatory protein, which regulates the transcription of several virulence factors, was expressed in many different treatments. As for 4125 isolate, the icaD regulator gene was positively influenced when different conditions were tested. In short, the data confirm that genes of S. aureus are expressed differently in response to concentrations of antibiotics below the considered inhibitory concentration and that the variance between strains of S. aureus makes difficult a pattern of expression is extended to all species.Staphylococcus aureus é um dos principais micro-organismos causadores da mastite bovina, doença que provoca as maiores perdas na pecuária leiteira mundial. Este patógeno possui diversos fatores de virulência que contribuem para a grande diversidade genética observada entre isolados e auxiliam no estabelecimento das infecções. Nos últimos anos, vários trabalhos têm demonstrado que o uso de antibióticos em concentrações subinibitórias modula a expressão gênica influenciando a virulência de patógenos bacterianos. Este trabalho teve por objetivo investigar o efeito de concentrações subinibitórias de ampicilina, gentamicina, oxacilina e tilosina, antibióticos usados em formulações veterinárias para o tratamento da mastite, na expressão de genes de duas cepas de S. aureus de origem bovina. Inicialmente, foi realizada uma investigação da presença de genes que codificam alguns fatores de virulência em 85 bactérias isoladas de animais com manifestação de mastite bovina. Os genes clfB e sdr foram os mais prevalentes, sendo detectados em 83,5% e 75,3% dos isolados, respectivamente. A diversidade genética dos isolados, avaliada por PCR multiplex, também foi alta e permitiu a discriminação de mais de 60 grupos. Esses resultados nortearam a escolha de S. aureus 4006 e 4125, com dissimilaridade genética de 80%, para os ensaios posteriores. RNA total das duas culturas crescidas em valores equivalentes a 0,5X, 0,25X e 0,125X da concentração inibitória mínima definida para cada antibióticos foi extraído e usado em análises de RT-PCR em tempo real. A expressão dos genes spa, clfB, sdrC, fnBP, icaD, icaR, murF e sarA foi normalizada para o gene gyrB. Todos os antibióticos testados causaram alteração na expressão dos genes avaliados. Um mesmo antibiótico usado em diferentes doses, assim como diferentes antibióticos em dose similar, modulou diferencialmente a expressão dos genes. Para o isolado 4006, a proteína regulatória SarA, que regula a transcrição de vários fatores de virulência, foi muita expressa em vários tratamentos. Já para o isolado 4125, o regulador icaD foi positivamente influenciado, quando diferentes condições foram testadas. Em suma, os dados confirmam que genes de S. aureus se expressam de forma diferenciada em resposta a concentrações de antibióticos abaixo das consideradas inibitórias e que a variação existente entre cepas de S. aureus dificulta que um padrão de expressão seja estendido a toda espécie.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaMestrado em Bioquímica AgrícolaUFVBRBioquímica e Biologia molecular de plantas; Bioquímica e Biologia molecular animalStaphylococcus aureusMastite bovinaStaphylococcus aureusBovine mastitisCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULARExpressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianosGene expression of Staphylococcus aureus isolated from bovine mastitis in response to subinhibitory concentrations of antibioticsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf3464377https://locus.ufv.br//bitstream/123456789/2426/1/texto%20completo.pdf051a57515bdffc736b40222c4b383acbMD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain137228https://locus.ufv.br//bitstream/123456789/2426/2/texto%20completo.pdf.txtf859836d5569e2e26a6af2c7a68286e8MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3567https://locus.ufv.br//bitstream/123456789/2426/3/texto%20completo.pdf.jpg9fea1719581381c039b2934cacddae02MD53123456789/24262016-04-08 23:02:52.53oai:locus.ufv.br:123456789/2426Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-09T02:02:52LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Expressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianos
dc.title.alternative.eng.fl_str_mv Gene expression of Staphylococcus aureus isolated from bovine mastitis in response to subinhibitory concentrations of antibiotics
title Expressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianos
spellingShingle Expressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianos
Klein, Raphael Contelli
Staphylococcus aureus
Mastite bovina
Staphylococcus aureus
Bovine mastitis
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR
title_short Expressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianos
title_full Expressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianos
title_fullStr Expressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianos
title_full_unstemmed Expressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianos
title_sort Expressão de genes de virulência de Staphylococcus aureus isolados de mastite bovina em resposta a concentrações subinibitórias de antimicrobianos
author Klein, Raphael Contelli
author_facet Klein, Raphael Contelli
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/7185650565356570
dc.contributor.author.fl_str_mv Klein, Raphael Contelli
dc.contributor.advisor-co1.fl_str_mv Bazzolli, Denise Mara Soares
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4761710D6
dc.contributor.advisor-co2.fl_str_mv Fietto, Luciano Gomes
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763824H8
dc.contributor.advisor1.fl_str_mv Ribon, Andréa de Oliveira Barros
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026E6
dc.contributor.referee1.fl_str_mv Zerbini, Poliane Alfenas
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/8632328159533071
dc.contributor.referee2.fl_str_mv Fietto, Juliana Lopes Rangel
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790238D0
contributor_str_mv Bazzolli, Denise Mara Soares
Fietto, Luciano Gomes
Ribon, Andréa de Oliveira Barros
Zerbini, Poliane Alfenas
Fietto, Juliana Lopes Rangel
dc.subject.por.fl_str_mv Staphylococcus aureus
Mastite bovina
topic Staphylococcus aureus
Mastite bovina
Staphylococcus aureus
Bovine mastitis
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR
dc.subject.eng.fl_str_mv Staphylococcus aureus
Bovine mastitis
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR
description Staphylococcus aureus is one of the main micro-organisms causing bovine mastitis, a disease that causes the greatest losses in dairy farming worldwide. This pathogen has several virulence factors that contribute to the great genetic diversity observed among isolates and assist in the establishment of infections. In recent years, several studies have shown that the use of antibiotics in subinhibitory concentrations modulates gene expression, influencing the virulence of bacterial pathogens. This study aimed to investigate the effect of subinhibitory concentrations of ampicillin, gentamicin, oxacillin and tylosin, antibiotics used in veterinary formulations for the treatment of mastitis, in the expression of genes from two strains of S. aureus of bovine origin. Initially, we conducted a preliminary investigation of the presence of genes encoding some virulence factors in 85 bacteria isolated from animals with bovine mastitis outbreak. Genes clfB and sdrCDE were the most prevalent, detected in 83.5% and 75.3% of the isolates, respectively. The genetic diversity of isolates, assessed by multiplex PCR, was also high and allowed the discrimination of more than 60 groups. These results guided the choice of S. aureus 4006 and 4125, with genetic similarity of 80% for the later trials. Total RNA from two cultures grown in values equivalent to 0.5X, 0.25X, 0.125X and the minimum inhibitory concentration defined for the four antibiotics was extracted and used in the analysis of real time RT-PCR. The expression of genes spa, clfB, sdrC, fnBP, icaD, icaR, murf and sarA was normalized to the gyrB gene. All antibiotics tested caused alterations in the expression of genes evaluated. The same antibiotics used in different doses, as well as different antibiotics at the same dose, differentially modulate the expression of genes. Only for 4006, the SarA regulatory protein, which regulates the transcription of several virulence factors, was expressed in many different treatments. As for 4125 isolate, the icaD regulator gene was positively influenced when different conditions were tested. In short, the data confirm that genes of S. aureus are expressed differently in response to concentrations of antibiotics below the considered inhibitory concentration and that the variance between strains of S. aureus makes difficult a pattern of expression is extended to all species.
publishDate 2010
dc.date.issued.fl_str_mv 2010-02-23
dc.date.available.fl_str_mv 2011-10-21
2015-03-26T13:07:31Z
dc.date.accessioned.fl_str_mv 2015-03-26T13:07:31Z
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dc.identifier.citation.fl_str_mv KLEIN, Raphael Contelli. Gene expression of Staphylococcus aureus isolated from bovine mastitis in response to subinhibitory concentrations of antibiotics. 2010. 82 f. Dissertação (Mestrado em Bioquímica e Biologia molecular de plantas; Bioquímica e Biologia molecular animal) - Universidade Federal de Viçosa, Viçosa, 2010.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/2426
identifier_str_mv KLEIN, Raphael Contelli. Gene expression of Staphylococcus aureus isolated from bovine mastitis in response to subinhibitory concentrations of antibiotics. 2010. 82 f. Dissertação (Mestrado em Bioquímica e Biologia molecular de plantas; Bioquímica e Biologia molecular animal) - Universidade Federal de Viçosa, Viçosa, 2010.
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