Caracterização molecular e isolamentode clostrídios psicrofílicos e psicrotróficos associados à deterioração de carnes refrigeradas embaladas a vácuo

Detalhes bibliográficos
Ano de defesa: 2009
Autor(a) principal: BUENO, Cláudia Peixoto lattes
Orientador(a): MESQUITA, Albenones José de lattes
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Goiás
Programa de Pós-Graduação: Doutorado em Ciência Animal
Departamento: Ciências Agrárias
País: BR
Palavras-chave em Português:
PCR
Palavras-chave em Inglês:
PCR
Área do conhecimento CNPq:
Link de acesso: http://repositorio.bc.ufg.br/tede/handle/tde/1091
Resumo: The deterioration of vacuum packed refrigerated meat accompanied by large gas production - a phenomenon called blown pack - is considered a major cause of economic losses of the meat industry in several regions of Brazil and the world. Several psychrophilic and psychrotrophic microorganisms may be involved, especially species of Clostridium. The objective of the present study was to perform the molecular characterization, through the use of the PCR technique, and the molecular isolation by conventional bacteriology, of the main microorganisms that cause blown pack in refrigerated meat from Brazil, New Zealand and the United Kingdom. Thus, typing techniques were used to differentiate the species and subspecies involved in this type of deterioration. Tirty-six samples of Brazilian blown pack meat, 6 samples from the UK and 12 experimental blown pack samples of venison from the North Island of New Zealand were analyzed. Three pairs of primers, the RFP / RRP, the 16SEF/16SER and the pair EISRF / EISRR were used for C. estertheticum estertheticum and Clostridium estertheticum like, and one for C. gasigenes (16DBF/16DBR). The samples with the PCR results were sent to a microbiology laboratory for conventional isolation of Clostridium estertheticum. It was concluded that Clostridium estertheticum estertheticum is responsible for the deterioration of meat and hence the blown pack in the UK. Samples of blown pack Brazilian meat have Clostridium estertheticum like as primary causal agent. The typing was carried out in isolates and strains - donated by Mirinz Center / Ruakura Agresearch / Hamilton / New Zealand - together with two isolates from Brazil, involved in this type of deterioration. The selected techniques AFLP and RFLP - PCR were able to distinguish species and subspecies of psychrophilic and psychrotrophic clostridia. However, the AFLP showed the highest discriminatory power, being able to distinguish 100% of the species - C. estertheticum, C. frigoris, C. bowmani, C. lacusfryxellense and C. psychrophylum - and also the subspecies C. estertheticum estertheticum, C. estertheticum laramiense, C. estertheticum like k21 and k24. Through the technique of RFLP, it was possible to differentiate the species of clostridia psychrotrophic, psichrophilic and also the subspecies C. estertheticum estertheticum and Clostridium estertheticum like, along with the use of four restriction endonucleases - AluI, CfoI, TaqI and HaeIII. The HaeIII provided greater variety of fragments and the ability to differentiate the species of clostridia psychrophilic and psychrotrophic, whereas TaqI was the only enzyme capable of differentiating the subspecies of C. estertheticum estertheticum and C. estertheticum laramiense of C. estertheticum like. The Brazilian samples isolated fit into the group of Clostridium estertheticum like, although there is no confirmation of the absence of Clostridium estertheticum estertheticum in the country.
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spelling MESQUITA, Albenones José dehttp://lattes.cnpq.br/3180029815183858PRADO, Cristiano Saleshttp://lattes.cnpq.br/1661902818272650http://lattes.cnpq.br/0790654390056879BUENO, Cláudia Peixoto2014-07-29T15:13:39Z2010-10-252009-02-20BUENO, Cláudia Peixoto. Molecular characterization and psychrophilic and psychrotrophic clostridia isolamentode associated with deterioration of chilled vacuum packed. 2009. 103 f. Tese (Doutorado em Ciências Agrárias) - Universidade Federal de Goiás, Goiânia, 2009.http://repositorio.bc.ufg.br/tede/handle/tde/1091The deterioration of vacuum packed refrigerated meat accompanied by large gas production - a phenomenon called blown pack - is considered a major cause of economic losses of the meat industry in several regions of Brazil and the world. Several psychrophilic and psychrotrophic microorganisms may be involved, especially species of Clostridium. The objective of the present study was to perform the molecular characterization, through the use of the PCR technique, and the molecular isolation by conventional bacteriology, of the main microorganisms that cause blown pack in refrigerated meat from Brazil, New Zealand and the United Kingdom. Thus, typing techniques were used to differentiate the species and subspecies involved in this type of deterioration. Tirty-six samples of Brazilian blown pack meat, 6 samples from the UK and 12 experimental blown pack samples of venison from the North Island of New Zealand were analyzed. Three pairs of primers, the RFP / RRP, the 16SEF/16SER and the pair EISRF / EISRR were used for C. estertheticum estertheticum and Clostridium estertheticum like, and one for C. gasigenes (16DBF/16DBR). The samples with the PCR results were sent to a microbiology laboratory for conventional isolation of Clostridium estertheticum. It was concluded that Clostridium estertheticum estertheticum is responsible for the deterioration of meat and hence the blown pack in the UK. Samples of blown pack Brazilian meat have Clostridium estertheticum like as primary causal agent. The typing was carried out in isolates and strains - donated by Mirinz Center / Ruakura Agresearch / Hamilton / New Zealand - together with two isolates from Brazil, involved in this type of deterioration. The selected techniques AFLP and RFLP - PCR were able to distinguish species and subspecies of psychrophilic and psychrotrophic clostridia. However, the AFLP showed the highest discriminatory power, being able to distinguish 100% of the species - C. estertheticum, C. frigoris, C. bowmani, C. lacusfryxellense and C. psychrophylum - and also the subspecies C. estertheticum estertheticum, C. estertheticum laramiense, C. estertheticum like k21 and k24. Through the technique of RFLP, it was possible to differentiate the species of clostridia psychrotrophic, psichrophilic and also the subspecies C. estertheticum estertheticum and Clostridium estertheticum like, along with the use of four restriction endonucleases - AluI, CfoI, TaqI and HaeIII. The HaeIII provided greater variety of fragments and the ability to differentiate the species of clostridia psychrophilic and psychrotrophic, whereas TaqI was the only enzyme capable of differentiating the subspecies of C. estertheticum estertheticum and C. estertheticum laramiense of C. estertheticum like. The Brazilian samples isolated fit into the group of Clostridium estertheticum like, although there is no confirmation of the absence of Clostridium estertheticum estertheticum in the country.A deterioração de carnes refrigeradas embaladas a vácuo acompanhada de grande produção de gás - fenômeno denominado tufamento de embalagens - é considerada uma das principais causas de perdas econômicas da indústria cárnea, em várias regiões do Brasil e do mundo. Diversos microrganismos psicrofílicos e psicrotróficos podem estar envolvidos, destacando-se espécies do gênero Clostridium. Objetivou-se com o presente estudo a caracterização molecular, por meio do emprego da técnica de PCR e o isolamento pela bacteriologia convencional, dos principais microrganismos causadores do tufamento de embalagens de carnes refrigeradas procedentes do Brasil, Nova Zelândia e Reino Unido. Para tanto, foram empregadas técnicas de tipagem visando a diferenciação das espécies e subespécies envolvidas neste tipo de deterioração. Foram analisadas 36 amostras tufadas de carnes brasileiras, 6 amostras tufadas oriundas do Reino Unido e 12 amostras experimentais tufadas de carne de cervo provenientes da Ilha Norte da Nova Zelândia. Foram utilizados três pares de primers, o RFP/RRP, o 16SEF/16SER e o par EISRF/EISRR para C. estertheticum estertheticum e Clostridium estertheticum like e um para C. gasigenes (16DBF/16DBR). As amostras com os resultados positivos ao PCR foram enviadas para a microbiologia convencional para isolamento do Clostridium estertheticum. Concluiu-se que o Clostridium estertheticum estertheticum é responsável pela deterioração das carnes e, consequentemente, pelo tufamento das embalagens no Reino Unido. As amostras tufadas de embalagens de carnes brasileiras têm como principal agente causador o Clostridium estertheticum like. A tipagem foi relizada em cepas e isolados doados pelo Mirinz Centre/ Ruakura AgResearch/ Hamilton/NZ - juntamente a dois isolados brasileiros, envolvidos nesse tipo de deterioração . As técnicas eleitas AFLP e RFLP PCR foram capazes de distinguir as espécies e subespécies de clostrídios psicrofílicos e psicrotróficos, porém, o AFLP apresentou maior poder discriminatório, sendo capaz de diferenciar 100% das espécies C. estertheticum, C. frigoris, C. bowmani, C. lacusfryxellense e C psychrophylum - e também as subespécies - Clostridium estertheticum estertheticum, Clostridium estertheticum laramiense, o Clostridium estertheticum like K21 e Clostridium estertheticum like K24. Por meio da técnica de RFLP foi possível diferenciar as espécies do clostrídio psicrofílicos e psicrotróficos e também as subespécies C. estertheticum estertheticum e Clostridium estertheticum like, utilizando em conjunto as quatro endonucleases de restrição AluI, CfoI, TaqI e HaeIII. A HaeIII proporciona maior variedade de fragmentos e capacidade de diferenciar as espécies de clostrídios psicrofílicos e psicrotróficos. Já a TaqI foi a única enzima capaz de diferenciar as subespécies C. estertheticum estertheticum e C. estertheticum laramiense do C. estertheticum like. As amostras brasileiras isoladas se enquadraram no grupo do Clostridium estertheticum like.Made available in DSpace on 2014-07-29T15:13:39Z (GMT). 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dc.title.por.fl_str_mv Caracterização molecular e isolamentode clostrídios psicrofílicos e psicrotróficos associados à deterioração de carnes refrigeradas embaladas a vácuo
dc.title.alternative.eng.fl_str_mv Molecular characterization and psychrophilic and psychrotrophic clostridia isolamentode associated with deterioration of chilled vacuum packed
title Caracterização molecular e isolamentode clostrídios psicrofílicos e psicrotróficos associados à deterioração de carnes refrigeradas embaladas a vácuo
spellingShingle Caracterização molecular e isolamentode clostrídios psicrofílicos e psicrotróficos associados à deterioração de carnes refrigeradas embaladas a vácuo
BUENO, Cláudia Peixoto
bacteriologia
carne
contaminação bacteriana
PCR
tipagem
tufamento de embalagem
vácuo
1.Clostrídios 2.Carnes-Refrigeração 3.Carne Qualidade
bacterial contamination
bacteriology
blown pack
meat
PCR
typing
vacuum
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::TECNOLOGIA DE ALIMENTOS
title_short Caracterização molecular e isolamentode clostrídios psicrofílicos e psicrotróficos associados à deterioração de carnes refrigeradas embaladas a vácuo
title_full Caracterização molecular e isolamentode clostrídios psicrofílicos e psicrotróficos associados à deterioração de carnes refrigeradas embaladas a vácuo
title_fullStr Caracterização molecular e isolamentode clostrídios psicrofílicos e psicrotróficos associados à deterioração de carnes refrigeradas embaladas a vácuo
title_full_unstemmed Caracterização molecular e isolamentode clostrídios psicrofílicos e psicrotróficos associados à deterioração de carnes refrigeradas embaladas a vácuo
title_sort Caracterização molecular e isolamentode clostrídios psicrofílicos e psicrotróficos associados à deterioração de carnes refrigeradas embaladas a vácuo
author BUENO, Cláudia Peixoto
author_facet BUENO, Cláudia Peixoto
author_role author
dc.contributor.advisor1.fl_str_mv MESQUITA, Albenones José de
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/3180029815183858
dc.contributor.advisor-co1.fl_str_mv PRADO, Cristiano Sales
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/1661902818272650
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/0790654390056879
dc.contributor.author.fl_str_mv BUENO, Cláudia Peixoto
contributor_str_mv MESQUITA, Albenones José de
PRADO, Cristiano Sales
dc.subject.por.fl_str_mv bacteriologia
carne
contaminação bacteriana
PCR
tipagem
tufamento de embalagem
vácuo
1.Clostrídios 2.Carnes-Refrigeração 3.Carne Qualidade
topic bacteriologia
carne
contaminação bacteriana
PCR
tipagem
tufamento de embalagem
vácuo
1.Clostrídios 2.Carnes-Refrigeração 3.Carne Qualidade
bacterial contamination
bacteriology
blown pack
meat
PCR
typing
vacuum
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::TECNOLOGIA DE ALIMENTOS
dc.subject.eng.fl_str_mv bacterial contamination
bacteriology
blown pack
meat
PCR
typing
vacuum
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::TECNOLOGIA DE ALIMENTOS
description The deterioration of vacuum packed refrigerated meat accompanied by large gas production - a phenomenon called blown pack - is considered a major cause of economic losses of the meat industry in several regions of Brazil and the world. Several psychrophilic and psychrotrophic microorganisms may be involved, especially species of Clostridium. The objective of the present study was to perform the molecular characterization, through the use of the PCR technique, and the molecular isolation by conventional bacteriology, of the main microorganisms that cause blown pack in refrigerated meat from Brazil, New Zealand and the United Kingdom. Thus, typing techniques were used to differentiate the species and subspecies involved in this type of deterioration. Tirty-six samples of Brazilian blown pack meat, 6 samples from the UK and 12 experimental blown pack samples of venison from the North Island of New Zealand were analyzed. Three pairs of primers, the RFP / RRP, the 16SEF/16SER and the pair EISRF / EISRR were used for C. estertheticum estertheticum and Clostridium estertheticum like, and one for C. gasigenes (16DBF/16DBR). The samples with the PCR results were sent to a microbiology laboratory for conventional isolation of Clostridium estertheticum. It was concluded that Clostridium estertheticum estertheticum is responsible for the deterioration of meat and hence the blown pack in the UK. Samples of blown pack Brazilian meat have Clostridium estertheticum like as primary causal agent. The typing was carried out in isolates and strains - donated by Mirinz Center / Ruakura Agresearch / Hamilton / New Zealand - together with two isolates from Brazil, involved in this type of deterioration. The selected techniques AFLP and RFLP - PCR were able to distinguish species and subspecies of psychrophilic and psychrotrophic clostridia. However, the AFLP showed the highest discriminatory power, being able to distinguish 100% of the species - C. estertheticum, C. frigoris, C. bowmani, C. lacusfryxellense and C. psychrophylum - and also the subspecies C. estertheticum estertheticum, C. estertheticum laramiense, C. estertheticum like k21 and k24. Through the technique of RFLP, it was possible to differentiate the species of clostridia psychrotrophic, psichrophilic and also the subspecies C. estertheticum estertheticum and Clostridium estertheticum like, along with the use of four restriction endonucleases - AluI, CfoI, TaqI and HaeIII. The HaeIII provided greater variety of fragments and the ability to differentiate the species of clostridia psychrophilic and psychrotrophic, whereas TaqI was the only enzyme capable of differentiating the subspecies of C. estertheticum estertheticum and C. estertheticum laramiense of C. estertheticum like. The Brazilian samples isolated fit into the group of Clostridium estertheticum like, although there is no confirmation of the absence of Clostridium estertheticum estertheticum in the country.
publishDate 2009
dc.date.issued.fl_str_mv 2009-02-20
dc.date.available.fl_str_mv 2010-10-25
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dc.identifier.citation.fl_str_mv BUENO, Cláudia Peixoto. Molecular characterization and psychrophilic and psychrotrophic clostridia isolamentode associated with deterioration of chilled vacuum packed. 2009. 103 f. Tese (Doutorado em Ciências Agrárias) - Universidade Federal de Goiás, Goiânia, 2009.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tde/1091
identifier_str_mv BUENO, Cláudia Peixoto. Molecular characterization and psychrophilic and psychrotrophic clostridia isolamentode associated with deterioration of chilled vacuum packed. 2009. 103 f. Tese (Doutorado em Ciências Agrárias) - Universidade Federal de Goiás, Goiânia, 2009.
url http://repositorio.bc.ufg.br/tede/handle/tde/1091
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