Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.

Detalhes bibliográficos
Autor(a) principal: Soares, Bianca da Silva
Data de Publicação: 2013
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRRJ
Texto Completo: https://tede.ufrrj.br/jspui/handle/jspui/3470
Resumo: Staphylococcus aureus is the most important etiologic agent of bovine mastitis. This bacterial species presents genetic heterogeneity and has a characterized population by genetically diverse strains. The analysis of genetic variation is an important tool for epidemiological studies. In this study, we assessed the genetic diversity of 56 S.aureus isolated from bovine mastitic milk in seven properties in south region the state of Rio de Janeiro by ERICPCR and(GTG)5-PCR techniques. 12 protocols were tested by ERIC-PCR, selected three with higher discriminatory power. With protocol 01, was obtained 6 cluster with discriminatory power of 0,863. Observed the dispersion of isolates in different properties with genetic similarity and the presence of clonal strains circulating within the same state and properties indifferent cities. The protocol 02 was generated in order to improve the performance of the technique and provided to obtain 15 cluster with a discriminatory power of 0,961, where it was observed genetic diversity and variability between isolates. However, the protocol 02 showed no reproducibility. Thus, we tested a third protocol, obtained 16 cluster with a discriminatory power of 0,96. From this, also observe the dispersion of isolates in different properties with genetic similarity and the presence of clonal strains circulating in properties indifferent cities. To implement the technique (GTG)5-PCR were tested eight protocols, electing only one of higher discriminatory power. This protocol generated 15 clusters with a discriminatory power of 0.957.Observed the dispersion of isolates in different properties with genetic similarity and the presence of clonal strains circulating in properties indifferent cities. But subsequent attempts to apply the technique were not successful which indicates the need for adjustments in their standard in our laboratory in order to ensure that, once optimized, can generate reliable and reproducible results. The results were compared to prior a molecular typing by PFGE that generated six distinct genetic profiles in 19 isolates tested. Despite producing a high number of genetic profiles and good discriminatory power, these techniques showed low reproducibility, especially when compared with the PFGE technique.
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spelling Souza, Miliane Moreira Soares de010.761.987-32http://lattes.cnpq.br/0865211214618618Coelho, Shana de Mattos de OliveiraCoelho, Irene da SilvaPereira, Ingrid Annes060.162.377-01http://lattes.cnpq.br/7572115655465997Soares, Bianca da Silva2020-04-29T14:31:06Z2013-02-27SOARES, Bianca da Silva. Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.. 2013. 46 f. Disserta??o (Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.https://tede.ufrrj.br/jspui/handle/jspui/3470Staphylococcus aureus is the most important etiologic agent of bovine mastitis. This bacterial species presents genetic heterogeneity and has a characterized population by genetically diverse strains. The analysis of genetic variation is an important tool for epidemiological studies. In this study, we assessed the genetic diversity of 56 S.aureus isolated from bovine mastitic milk in seven properties in south region the state of Rio de Janeiro by ERICPCR and(GTG)5-PCR techniques. 12 protocols were tested by ERIC-PCR, selected three with higher discriminatory power. With protocol 01, was obtained 6 cluster with discriminatory power of 0,863. Observed the dispersion of isolates in different properties with genetic similarity and the presence of clonal strains circulating within the same state and properties indifferent cities. The protocol 02 was generated in order to improve the performance of the technique and provided to obtain 15 cluster with a discriminatory power of 0,961, where it was observed genetic diversity and variability between isolates. However, the protocol 02 showed no reproducibility. Thus, we tested a third protocol, obtained 16 cluster with a discriminatory power of 0,96. From this, also observe the dispersion of isolates in different properties with genetic similarity and the presence of clonal strains circulating in properties indifferent cities. To implement the technique (GTG)5-PCR were tested eight protocols, electing only one of higher discriminatory power. This protocol generated 15 clusters with a discriminatory power of 0.957.Observed the dispersion of isolates in different properties with genetic similarity and the presence of clonal strains circulating in properties indifferent cities. But subsequent attempts to apply the technique were not successful which indicates the need for adjustments in their standard in our laboratory in order to ensure that, once optimized, can generate reliable and reproducible results. The results were compared to prior a molecular typing by PFGE that generated six distinct genetic profiles in 19 isolates tested. Despite producing a high number of genetic profiles and good discriminatory power, these techniques showed low reproducibility, especially when compared with the PFGE technique.O Staphylococcus aureus ? um importante agente causador de mastite bovina. Est? esp?cie bacteriana possui heterogeneidade gen?tica e uma popula??o caracterizada por estirpes geneticamente diversas. A an?lise da varia??o gen?tica ? uma importante ferramenta para fins de estudos epidemiol?gicos. No presente estudo, foi avaliada a diversidade gen?tica de 56 Staphylococcus aureus isolados de leite bovino mast?tico em sete propriedades da regi?o Sul- Fluminense do Rio de Janeiro atrav?s das t?cnicas de ERIC e (GTG)5-PCR. Foram testados 12 protocolos de ERIC-PCR, sendo selecionados tr?s com maior poder discriminat?rio. Com o protocolo 01, obteve-se 6 agrupamentos com um poder discriminat?rio de 0,863. Observou-se a dispers?o de isolados com similaridade gen?tica em diferentes propriedades e presen?a de cepas clonais circulantes dentro da mesma propriedade e em propriedades de munic?pios diferentes. O protocolo 02 foi gerado afim de melhorar a performance da t?cnica e propiciou a obten??o de15 agrupamentos com um poder discriminat?rio de 0,961, onde foi observada grande diversidade g?nica e variabilidade entre os isolados. No entanto, o protocolo 02 n?o apresentou reprodutibilidade. Desse modo, foi testado um terceiro protocolo, obtendo-se 16 agrupamentos com um poder discriminat?rio de 0,96. A partir deste, tamb?m observou-se a dispers?o de isolados com similaridade gen?tica em diferentes propriedades e presen?a de cepas clonais circulantes em propriedades de munic?pios diferentes. Para a execu??o da t?cnica de (GTG)5- PCR foram testados oito protocolos,elegendo-se apenas um de maior poder discriminat?rio.Este protocolo gerou 15 agrupamentos com um poder discriminat?rio de 0,957. Observou-se a dispers?o de isolados com similaridade gen?tica em diferentes propriedades e presen?a de cepas clonais circulantes em propriedades de munic?pios diferentes. Por?m as tentativas subsequentes de execu??o da t?cnica n?o foram bem sucedidas o que indica a necessidade de ajustes em sua padroniza??o em nosso laborat?rio de maneira a garantir que, uma vez otimizada, possa gerar resultados confi?veis e reprodut?veis. Os resultados obtidos foram comparados com uma tipagem molecular pr?via atrav?s do PFGE que gerou 6 perfis gen?ticos distintos em 19 isolados testados. Apesar de produzirem um n?mero elevado de perfis gen?ticos e um bom poder discriminat?rio, estas t?cnicas apresentaram pouca reprodutibilidade, especialmente quando comparadas com a t?cnica de PFGE.Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2020-04-29T14:31:06Z No. of bitstreams: 1 2013 - Bianca da Silva Soares.pdf: 964225 bytes, checksum: f4b16ad54dce12d47503fa9161391eff (MD5)Made available in DSpace on 2020-04-29T14:31:06Z (GMT). No. of bitstreams: 1 2013 - Bianca da Silva Soares.pdf: 964225 bytes, checksum: f4b16ad54dce12d47503fa9161391eff (MD5) Previous issue date: 2013-02-27Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior, CAPES, Brasil.application/pdfhttps://tede.ufrrj.br/retrieve/14486/2013%20-%20Bianca%20da%20Silva%20Soares.pdf.jpghttps://tede.ufrrj.br/retrieve/17500/2013%20-%20Bianca%20da%20Silva%20Soares.pdf.jpghttps://tede.ufrrj.br/retrieve/23820/2013%20-%20Bianca%20da%20Silva%20Soares.pdf.jpghttps://tede.ufrrj.br/retrieve/30204/2013%20-%20Bianca%20da%20Silva%20Soares.pdf.jpghttps://tede.ufrrj.br/retrieve/36500/2013%20-%20Bianca%20da%20Silva%20Soares.pdf.jpghttps://tede.ufrrj.br/retrieve/42868/2013%20-%20Bianca%20da%20Silva%20Soares.pdf.jpghttps://tede.ufrrj.br/retrieve/49326/2013%20-%20Bianca%20da%20Silva%20Soares.pdf.jpghttps://tede.ufrrj.br/retrieve/55742/2013%20-%20Bianca%20da%20Silva%20Soares.pdf.jpgporUniversidade Federal Rural do Rio de JaneiroPrograma de P?s-Gradua??o em Ci?ncias Veterin?riasUFRRJBrasilInstituto de Veterin?riaAARESTRUP F. 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dc.title.por.fl_str_mv Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.
dc.title.alternative.eng.fl_str_mv Use of REP-PCR for genetic diversity analyses of Staphylococcus aureus isolated from mastitis bovine.
title Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.
spellingShingle Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.
Soares, Bianca da Silva
mastite
Staphylococcus aureus
tipagem molecular
Mastitis,
Staphylococcus aureus
typing molecular
Ci?ncias Agr?rias
title_short Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.
title_full Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.
title_fullStr Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.
title_full_unstemmed Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.
title_sort Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.
author Soares, Bianca da Silva
author_facet Soares, Bianca da Silva
author_role author
dc.contributor.advisor1.fl_str_mv Souza, Miliane Moreira Soares de
dc.contributor.advisor1ID.fl_str_mv 010.761.987-32
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/0865211214618618
dc.contributor.advisor-co1.fl_str_mv Coelho, Shana de Mattos de Oliveira
dc.contributor.referee1.fl_str_mv Coelho, Irene da Silva
dc.contributor.referee2.fl_str_mv Pereira, Ingrid Annes
dc.contributor.authorID.fl_str_mv 060.162.377-01
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/7572115655465997
dc.contributor.author.fl_str_mv Soares, Bianca da Silva
contributor_str_mv Souza, Miliane Moreira Soares de
Coelho, Shana de Mattos de Oliveira
Coelho, Irene da Silva
Pereira, Ingrid Annes
dc.subject.por.fl_str_mv mastite
Staphylococcus aureus
tipagem molecular
topic mastite
Staphylococcus aureus
tipagem molecular
Mastitis,
Staphylococcus aureus
typing molecular
Ci?ncias Agr?rias
dc.subject.eng.fl_str_mv Mastitis,
Staphylococcus aureus
typing molecular
dc.subject.cnpq.fl_str_mv Ci?ncias Agr?rias
description Staphylococcus aureus is the most important etiologic agent of bovine mastitis. This bacterial species presents genetic heterogeneity and has a characterized population by genetically diverse strains. The analysis of genetic variation is an important tool for epidemiological studies. In this study, we assessed the genetic diversity of 56 S.aureus isolated from bovine mastitic milk in seven properties in south region the state of Rio de Janeiro by ERICPCR and(GTG)5-PCR techniques. 12 protocols were tested by ERIC-PCR, selected three with higher discriminatory power. With protocol 01, was obtained 6 cluster with discriminatory power of 0,863. Observed the dispersion of isolates in different properties with genetic similarity and the presence of clonal strains circulating within the same state and properties indifferent cities. The protocol 02 was generated in order to improve the performance of the technique and provided to obtain 15 cluster with a discriminatory power of 0,961, where it was observed genetic diversity and variability between isolates. However, the protocol 02 showed no reproducibility. Thus, we tested a third protocol, obtained 16 cluster with a discriminatory power of 0,96. From this, also observe the dispersion of isolates in different properties with genetic similarity and the presence of clonal strains circulating in properties indifferent cities. To implement the technique (GTG)5-PCR were tested eight protocols, electing only one of higher discriminatory power. This protocol generated 15 clusters with a discriminatory power of 0.957.Observed the dispersion of isolates in different properties with genetic similarity and the presence of clonal strains circulating in properties indifferent cities. But subsequent attempts to apply the technique were not successful which indicates the need for adjustments in their standard in our laboratory in order to ensure that, once optimized, can generate reliable and reproducible results. The results were compared to prior a molecular typing by PFGE that generated six distinct genetic profiles in 19 isolates tested. Despite producing a high number of genetic profiles and good discriminatory power, these techniques showed low reproducibility, especially when compared with the PFGE technique.
publishDate 2013
dc.date.issued.fl_str_mv 2013-02-27
dc.date.accessioned.fl_str_mv 2020-04-29T14:31:06Z
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.citation.fl_str_mv SOARES, Bianca da Silva. Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.. 2013. 46 f. Disserta??o (Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.
dc.identifier.uri.fl_str_mv https://tede.ufrrj.br/jspui/handle/jspui/3470
identifier_str_mv SOARES, Bianca da Silva. Aplica??o de T?cnicas de REP-PCR na An?lise de Isolados de Staphylococcus aureus em amostras de Mastite Bovina.. 2013. 46 f. Disserta??o (Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.
url https://tede.ufrrj.br/jspui/handle/jspui/3470
dc.language.iso.fl_str_mv por
language por
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