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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Emprego das técnicas de pcr e vidas na determinação da escherichia coli o157:H7 em queijos minas frescal em feiras livres e estabelecimentos sob inspeção federal / The use of PCR and VIDAS tchniques of Escherichia coli O157:H7 determination in Minas Frescal cheeses coleted from open air markets and establishments under Federal Inspection

CARVALHO, Rosangela Nunes 31 August 2009 (has links)
Made available in DSpace on 2014-07-29T15:07:52Z (GMT). No. of bitstreams: 1 rosangela.pdf: 1093995 bytes, checksum: 8b8d62975e344cdccd0d1777e48758b1 (MD5) Previous issue date: 2009-08-31 / Escherichia coli O157:H7 is a serotype that belongs to Shiga toxin-producing Escherichia coli group and it has been incriminated in outbreaks of food poisoning and also as cause of sporadic cases of hemorrhagic colitis and hemolytic uremic syndrome in many countries. In Brazil, there is no data available about outbreaks caused by E. coli O157:H7, but this pathogen has been found often in cattle feces. Taking into consideration the high susceptibility of Minas Frescal cheese to the contamination by E. coli O157:H7 and also considering that milk is frequently associated with outbreaks, the aim of this research was to determinate the occurrence of the pathogen in Minas Frescal cheese, and also to compare techniques for this bacterium detection, such as PCR (Polymerase Chain Reaction) and VIDAS ECO O157® (bioMérieux, Lyon, France) test. Thirty cheese samples were collected from open-air markets in Goiânia and thirty samples were collected from establishments under Federal Inspection located in Goiás that trade their products in supermarkets in Goiânia. The MPN (More probable number) of Thermo-tolerant Coliforms and MPN of E. coli were also determined from the samples. From those samples collected in open air markets, E. coli O157:H7 was detected in 6,67% and 23,33% when the technique used was VIDAS ECO O157® and PCR, respectively. From those samples collected in establishments under Federal Inspection, the bacterium was not detected. There was no significant difference (p>0,05) between VIDAS ECO O157® and PCR for pathogen detection in samples from open air market and establishments under Federal Inspection. No significant difference (p> 0,05) was observed in samples collected from open air market and industries under Federal Inspection when VIDAS ECO O157® was used to E.coli O157:H7 detection. When PCR technique was applied, there was significant difference (p< 0,05) between samples origin. In 86,67% of samples collected from open air markets, Thermo-tolerant Coliforms were detected over the level established in Brazilian legislation, but in samples collected from industries under Federal Inspection, this value was 10%. The Minas Frescal cheese samples collected from open air market were more contaminated by E.coli than those collected from establishments under Federal Inspection and commercialized in supermarkets. / Escherichia coli O157:H7 é um sorotipo de E.coli pertencente ao grupo STEC - E.coli produtora de toxina Shiga - que tem sido incriminado em surtos de toxinfecção alimentar bem como em casos esporádicos de colite hemorrágica e síndrome urêmica hemolítica em vários países. No Brasil não há dados sobre possíveis surtos causados por E. coli O157:H7, mas este patógeno vem sendo frequentemente encontrado em fezes bovinas. Tendo-se em vista a suscetibilidade do queijo Minas Frescal à contaminação por E.coli O157:H7 e que o leite é um dos alimentos mais envolvidos em surtos, objetivou com o presente trabalho determinar a ocorrência deste microrganismo em queijos Minas Frescal, além de comparar as técnicas de reação em cadeia da polimerase (PCR) e teste VIDAS ECO O157®(bioMérieux, Lyon, France) na detecção do patógeno. Para tanto, analisou-se 30 amostras oriundas de feiras livres de Goiânia e 30 de estabelecimentos sob Inspeção Federal localizados em Goiás e que comercializam seus produtos em supermercados de Goiânia. Também foram determinados o Número Mais Provável (NMP) de Coliformes Termotolerantes e NMP de E. coli nestas amostras. Nas amostras oriundas de feiras livres detectouse E.coli O157:H7 em 6,67% e 23,33% quando utilizou-se o VIDAS ECO O157® e PCR, espectivamente. Nas amostras provenientes de estabelecimentos sob Inspeção Federal o microrganismo não foi detectado. Não houve diferença significativa (p>0,05) entre VIDAS ECO O157® e PCR na detecção do patógeno em amostras oriundas de feiras livres e estabelecimentos sob Inspeção Federal. Não observou-se diferença significativa (p> 0,05) entre amostras oriundas de feiras livres e indústrias sob Inspeção Federal quando utilizou-se o VIDAS ECO O157® na detecção de E.coli O157:H7. Quando empregou-se a técnica de PCR, constatou-se diferença (p< 0,05) entre as origens das amostras. Em 86,67% das amostras oriundas de feiras livres detectou-se a presença de Coliformes Termotolerantes acima do nível estabelecido pela legislação enquanto que, nas amostras provenientes de estabelecimentos sob Inspeção Federal esse percentual foi de 10%. As amostras de queijo Minas Frescal provenientes de feiras livres revelaram-se mais contaminadas por E.coli do que as oriundas de estabelecimentos sob Inspeção Federal e comercializadas em supermercados.

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