Moléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leite

Detalhes bibliográficos
Autor(a) principal: Viana, Eliseth de Souza
Data de Publicação: 2006
Tipo de documento: Tese
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/1555
Resumo: This study aimed to evaluate the effect of different culture media on biofilm formation and proteolytic activity of sessile and planktonic cells of psychrotrophic bacteria isolated from cooled milk; to detect the presence of quorum sensing signaling molecules produced by these isolates and to evaluate the effects of furanones and acyl homoserines lactones (AHLs) on biofilm formation. The growth of Pseudomonas fluorescens was evaluated in polystyrene microplates containg different culture media. The proteolytic activity was determined using azocasein as substrate. The monitor strains Chromobacterium violaceum CV026, Agrobacterium tumefaciens A136, WCF47 and KYC55 and Escherichia coli pSB403 were used to study the production of quorum sensing signaling molecules produced by sessile and planktonic cells in agar diffusion assays and in thin layer chromatography (TLC). The effects of furanones and AHLs in biofilm formation were evaluated by spectrophotometry and scanning electronic microscopy. It was verified that complex media favored growth, but not biofilm formation by P. fluorescens strains. The salts minimal media MMS containing glycerol as carbon source enhanced biofilm formation by P. fluorescens in microplates and in stainless steel coupons and the strains 097, 0109, 0137 had higher numbers of sessile cells. Hafnia alvei 067, 071 and Aeromonas hydrophila 099 also adhered to coupons of stainless steel immersed in MMS, indicating the potential for these isolates to form biofilms. In MMS, the proteolytic activity of P. fluorescens 097 increased with the increase in viable cell number. The proteolytic activity was favored in tryptone yeast extract and phosphate media TYEP added with CaCl2. The evidence for quorum sensing signaling molecules production in P. fluorescens 097 was obtained by reduction of violacein production by C. violaceum and β-gal production by A. tumefaciens WCF47 in the presence of sessile cells and extracts from these cells. However, the presence of signaling molecules was not confirmed by TLC. The production of AHL was detected in biofims of H. alvei 059, 067, 071 and A. hydrophila 099, being A. tumefaciens WCF47 the monitor strain most sensible to AHLs. The extracts obtained from planktonic cells supernatants of H. alvei 071 induced positive responses in the monitors E. coli pSB403 and A. tumefaciens WCF47 in agar diffusion assays and in TLC. A significant difference was verified in the inhibition of biofilm formation by H. alvei 071 in the presence of furanones and scanning electronic microscopy images confirmed this result. The addition of AHLs did not affect the biofilm formation in the strains evaluated. The detection of signaling molecules produced by sessile cells of H. alvei strains and the inhibitory effect of furanones suggest that quorum sensing influences the regulation of these structures. These results contributed to better understand the mechanisms involved in the regulation of biofilm formation by bacteria isolated from milk and it can also contribute to propose new strategies to prevent and control biofilm formation.
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spelling Viana, Eliseth de Souzahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4778765P8Mantovani, Hilário Cuquettohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026Z7Tótola, Marcos Rogériohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727020U4Vanetti, Maria Cristina Dantashttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783874H3Alves, Virgínia Maria Chaveshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781053Z0Pinto, Cláudia Lúcia de Oliveirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783521J62015-03-26T12:50:58Z2007-02-062015-03-26T12:50:58Z2006-12-12VIANA, Eliseth de Souza. Quorum sensing signaling molecules in biofilms formed by psychrotrophic bacteria isolated from milk. 2006. 176 f. Tese (Doutorado em Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse) - Universidade Federal de Viçosa, Viçosa, 2006.http://locus.ufv.br/handle/123456789/1555This study aimed to evaluate the effect of different culture media on biofilm formation and proteolytic activity of sessile and planktonic cells of psychrotrophic bacteria isolated from cooled milk; to detect the presence of quorum sensing signaling molecules produced by these isolates and to evaluate the effects of furanones and acyl homoserines lactones (AHLs) on biofilm formation. The growth of Pseudomonas fluorescens was evaluated in polystyrene microplates containg different culture media. The proteolytic activity was determined using azocasein as substrate. The monitor strains Chromobacterium violaceum CV026, Agrobacterium tumefaciens A136, WCF47 and KYC55 and Escherichia coli pSB403 were used to study the production of quorum sensing signaling molecules produced by sessile and planktonic cells in agar diffusion assays and in thin layer chromatography (TLC). The effects of furanones and AHLs in biofilm formation were evaluated by spectrophotometry and scanning electronic microscopy. It was verified that complex media favored growth, but not biofilm formation by P. fluorescens strains. The salts minimal media MMS containing glycerol as carbon source enhanced biofilm formation by P. fluorescens in microplates and in stainless steel coupons and the strains 097, 0109, 0137 had higher numbers of sessile cells. Hafnia alvei 067, 071 and Aeromonas hydrophila 099 also adhered to coupons of stainless steel immersed in MMS, indicating the potential for these isolates to form biofilms. In MMS, the proteolytic activity of P. fluorescens 097 increased with the increase in viable cell number. The proteolytic activity was favored in tryptone yeast extract and phosphate media TYEP added with CaCl2. The evidence for quorum sensing signaling molecules production in P. fluorescens 097 was obtained by reduction of violacein production by C. violaceum and β-gal production by A. tumefaciens WCF47 in the presence of sessile cells and extracts from these cells. However, the presence of signaling molecules was not confirmed by TLC. The production of AHL was detected in biofims of H. alvei 059, 067, 071 and A. hydrophila 099, being A. tumefaciens WCF47 the monitor strain most sensible to AHLs. The extracts obtained from planktonic cells supernatants of H. alvei 071 induced positive responses in the monitors E. coli pSB403 and A. tumefaciens WCF47 in agar diffusion assays and in TLC. A significant difference was verified in the inhibition of biofilm formation by H. alvei 071 in the presence of furanones and scanning electronic microscopy images confirmed this result. The addition of AHLs did not affect the biofilm formation in the strains evaluated. The detection of signaling molecules produced by sessile cells of H. alvei strains and the inhibitory effect of furanones suggest that quorum sensing influences the regulation of these structures. These results contributed to better understand the mechanisms involved in the regulation of biofilm formation by bacteria isolated from milk and it can also contribute to propose new strategies to prevent and control biofilm formation.Os objetivos deste trabalho foram avaliar o efeito de diferentes meios de cultivo na formação de biofilmes e na atividade proteolítica de células sésseis e planctônicas de bactérias psicrotróficas isoladas de leite refrigerado; detectar a presença de moléculas sinalizadoras de quorum sensing produzidas por esses isolados e avaliar o efeito de furanonas e de acil homoserinas lactonas (AHLs) na formação de biofilmes. O crescimento de Pseudomonas fluorescens foi acompanhado em microplacas de poliestireno contendo diferentes meios de cultivo. A atividade proteolítica foi determinada, utilizando-se a azocaseína como substrato. As estirpes monitoras Chromobacterium violaceum CV026, Agrobacterium tumefaciens A136, WCF47 e KYC55 e Escherichia coli pSB403 foram utilizadas para estudar a produção de moléculas sinalizadoras de quorum sensing em células sésseis e planctônicas em ensaios de difusão em meios de cultura e em cromatografia em camada delgada (CCD). O efeito de furanonas e de AHLs na formação de biofilmes foi avaliado por espectrofotometria e por microscopia eletrônica de varredura. Verificou-se que meios complexos favoreceram o crescimento, mas não a formação dos biofilmes por estirpes de P. fluorescens. O cultivo em meio mínimo de sais MMS, contendo glicerol como fonte de carbono, aumentou a formação de biofilmes de P. fluorescens em microplacas e em cupons de aço inoxidável e os isolados 097, 0109, 0137 e 0141 destacaram-se por apresentar maior número de células sésseis. Isolados de Hafnia alvei 067, 071 e Aeromonas hydrophila 099 também aderiram aos cupons de aço inoxidável imersos nos meios MMS, indicando o potencial destes isolados para formar biofilme. Em MMS, a atividade proteolítica da estirpe 097 de P. fluorescens aumentou com o aumento do número de células viáveis e foi favorecida em meio triptona, extrato de levedura e fosfato - TYEP adicionado de CaCl2. A indicação da produção de moléculas sinalizadoras de quorum sensing em biofilmes de P. fluorescens 097 foi obtida pela redução da produção de violaceína pela estirpe C. violaceum e pelo estímulo da produção de ß-gal por A. tumefaciens WCF47 na presença de células sésseis e de extratos dessas células. Entretanto, a presença de moléculas sinalizadoras em extratos de células sésseis e planctônicas de P. fluorescens não foi confirmada pela técnica de CCD. A produção de AHLs foi detectada nos biofilmes de H. alvei 059, 067, 071 e A. hydrophila 099, sendo A. tumefaciens WCF47 a estirpe monitora mais sensível. Os extratos obtidos a partir de células sésseis e planctônicas de H. alvei 071 induziram resposta positiva nas monitoras E. coli pSB403 e A. tumefaciens WCF47 nos ensaios de difusão em ágar e em cromatografia em camada delgada. Verificou-se diferença significativa na inibição da formação de biofilmes de H. alvei 071 pelas furanonas, e este resultado foi confirmado por imagens obtidas por microscopia eletrônica de varredura. A adição de AHLs não afetou a formação de biofilmes nas estirpes avaliadas. A detecção de moléculas sinalizadoras produzidas pelas células sésseis das estirpes de H. alvei avaliadas e o efeito inibidor da adição de furanonas na formação dos biofilmes sugeriram a participação do sistema quorum sensing na regulação da formação dessas estruturas. Os resultados obtidos contribuem para melhor entendimento de mecanismos envolvidos na regulação da formação de biofilmes por bactérias isoladas de leite e servirão para a elaboração de estratégias de prevenção e controle de sua formação.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaDoutorado em Microbiologia AgrícolaUFVBRAssociações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesseBactérias psicrotróficasBiofilmeQuorum sensingLeitePsychrotrophic bacteriaBiofilmQuorum sensingMilkCNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS::MICROBIOLOGIA DE ALIMENTOSMoléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leiteQuorum sensing signaling molecules in biofilms formed by psychrotrophic bacteria isolated from milkinfo: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/pdf3528301https://locus.ufv.br//bitstream/123456789/1555/1/texto%20completo.pdfed004aa5c4b14da113d412ade0d695aeMD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain335627https://locus.ufv.br//bitstream/123456789/1555/2/texto%20completo.pdf.txtc5559240055dcdc715ce0c3014ab0ac3MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3814https://locus.ufv.br//bitstream/123456789/1555/3/texto%20completo.pdf.jpg80c3543d8a519855e6b00de20acdd8f8MD53123456789/15552016-04-07 23:05:06.894oai:locus.ufv.br:123456789/1555Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-08T02:05:06LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Moléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leite
dc.title.alternative.eng.fl_str_mv Quorum sensing signaling molecules in biofilms formed by psychrotrophic bacteria isolated from milk
title Moléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leite
spellingShingle Moléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leite
Viana, Eliseth de Souza
Bactérias psicrotróficas
Biofilme
Quorum sensing
Leite
Psychrotrophic bacteria
Biofilm
Quorum sensing
Milk
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS::MICROBIOLOGIA DE ALIMENTOS
title_short Moléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leite
title_full Moléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leite
title_fullStr Moléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leite
title_full_unstemmed Moléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leite
title_sort Moléculas sinalizadoras de quorum sensing em biofilmes formados por bactérias psicrotróficas isoladas de leite
author Viana, Eliseth de Souza
author_facet Viana, Eliseth de Souza
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4778765P8
dc.contributor.author.fl_str_mv Viana, Eliseth de Souza
dc.contributor.advisor-co1.fl_str_mv Mantovani, Hilário Cuquetto
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026Z7
dc.contributor.advisor-co2.fl_str_mv Tótola, Marcos Rogério
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727020U4
dc.contributor.advisor1.fl_str_mv Vanetti, Maria Cristina Dantas
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783874H3
dc.contributor.referee1.fl_str_mv Alves, Virgínia Maria Chaves
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781053Z0
dc.contributor.referee2.fl_str_mv Pinto, Cláudia Lúcia de Oliveira
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783521J6
contributor_str_mv Mantovani, Hilário Cuquetto
Tótola, Marcos Rogério
Vanetti, Maria Cristina Dantas
Alves, Virgínia Maria Chaves
Pinto, Cláudia Lúcia de Oliveira
dc.subject.por.fl_str_mv Bactérias psicrotróficas
Biofilme
Quorum sensing
Leite
topic Bactérias psicrotróficas
Biofilme
Quorum sensing
Leite
Psychrotrophic bacteria
Biofilm
Quorum sensing
Milk
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS::MICROBIOLOGIA DE ALIMENTOS
dc.subject.eng.fl_str_mv Psychrotrophic bacteria
Biofilm
Quorum sensing
Milk
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS::MICROBIOLOGIA DE ALIMENTOS
description This study aimed to evaluate the effect of different culture media on biofilm formation and proteolytic activity of sessile and planktonic cells of psychrotrophic bacteria isolated from cooled milk; to detect the presence of quorum sensing signaling molecules produced by these isolates and to evaluate the effects of furanones and acyl homoserines lactones (AHLs) on biofilm formation. The growth of Pseudomonas fluorescens was evaluated in polystyrene microplates containg different culture media. The proteolytic activity was determined using azocasein as substrate. The monitor strains Chromobacterium violaceum CV026, Agrobacterium tumefaciens A136, WCF47 and KYC55 and Escherichia coli pSB403 were used to study the production of quorum sensing signaling molecules produced by sessile and planktonic cells in agar diffusion assays and in thin layer chromatography (TLC). The effects of furanones and AHLs in biofilm formation were evaluated by spectrophotometry and scanning electronic microscopy. It was verified that complex media favored growth, but not biofilm formation by P. fluorescens strains. The salts minimal media MMS containing glycerol as carbon source enhanced biofilm formation by P. fluorescens in microplates and in stainless steel coupons and the strains 097, 0109, 0137 had higher numbers of sessile cells. Hafnia alvei 067, 071 and Aeromonas hydrophila 099 also adhered to coupons of stainless steel immersed in MMS, indicating the potential for these isolates to form biofilms. In MMS, the proteolytic activity of P. fluorescens 097 increased with the increase in viable cell number. The proteolytic activity was favored in tryptone yeast extract and phosphate media TYEP added with CaCl2. The evidence for quorum sensing signaling molecules production in P. fluorescens 097 was obtained by reduction of violacein production by C. violaceum and β-gal production by A. tumefaciens WCF47 in the presence of sessile cells and extracts from these cells. However, the presence of signaling molecules was not confirmed by TLC. The production of AHL was detected in biofims of H. alvei 059, 067, 071 and A. hydrophila 099, being A. tumefaciens WCF47 the monitor strain most sensible to AHLs. The extracts obtained from planktonic cells supernatants of H. alvei 071 induced positive responses in the monitors E. coli pSB403 and A. tumefaciens WCF47 in agar diffusion assays and in TLC. A significant difference was verified in the inhibition of biofilm formation by H. alvei 071 in the presence of furanones and scanning electronic microscopy images confirmed this result. The addition of AHLs did not affect the biofilm formation in the strains evaluated. The detection of signaling molecules produced by sessile cells of H. alvei strains and the inhibitory effect of furanones suggest that quorum sensing influences the regulation of these structures. These results contributed to better understand the mechanisms involved in the regulation of biofilm formation by bacteria isolated from milk and it can also contribute to propose new strategies to prevent and control biofilm formation.
publishDate 2006
dc.date.issued.fl_str_mv 2006-12-12
dc.date.available.fl_str_mv 2007-02-06
2015-03-26T12:50:58Z
dc.date.accessioned.fl_str_mv 2015-03-26T12:50:58Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
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dc.identifier.citation.fl_str_mv VIANA, Eliseth de Souza. Quorum sensing signaling molecules in biofilms formed by psychrotrophic bacteria isolated from milk. 2006. 176 f. Tese (Doutorado em Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse) - Universidade Federal de Viçosa, Viçosa, 2006.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/1555
identifier_str_mv VIANA, Eliseth de Souza. Quorum sensing signaling molecules in biofilms formed by psychrotrophic bacteria isolated from milk. 2006. 176 f. Tese (Doutorado em Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse) - Universidade Federal de Viçosa, Viçosa, 2006.
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dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse
publisher.none.fl_str_mv Universidade Federal de Viçosa
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