Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo

Detalhes bibliográficos
Autor(a) principal: Costa, Ana Carolina Cabral Carvalhaes
Data de Publicação: 2016
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFG
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/6628
Resumo: Due to their antagonistic activity against undesirable microorganisms, lactic acid bacteria (LAB) are a promising alternative for conservation and improvement of food safety. The aim of this work was isolate, from fresh Minas cheese, bacteriocin producing LAB with potential for use in biopreservation systems of dairy products. For this purpose, cheese samples were evaluated at the beginning and end of their shelf life and the isolation was performed by surface plating on MRS agar. The antimicrobial activity was verified by antagonism assay using Listeria monocytogenes ATCC 7644 and Staphylococcus aureus ATCC 25923 as indicators microorganisms. The selected LAB were submitted to the spot-on-the-lawn test, Gram stain, catalase and oxidase production and glucose metabolism in Hugh & Leifson broth. The occurrence of inhibition due to the production of organic acids and bacteriophages was discarded. The protein nature of antagonistic substances was confirmed by using proteases. After the well-diffusion assay, four LAB with satisfactory antagonistic activity against the indicators microorganisms were selected for identification by sequencing of the 16S RNA gene. All bacteria were identified as Lactococcus lactis. Based on antagonism assay, Lactococcus lactis QMF 11 was selected for use in co-inoculation studies with pathogens in pasteurized milk, kept at 8°C for 10 days. Lactobacillus sakei ATCC 15521 was used as negative control for bacteriocin production. After incubation period, monocultures of Listeria monocytogenes reached 8 log cfu mL-1 and in the presence of Lactobacillus sakei ATCC its population achieved 7.3 log cfu mL-1. However, when co-inoculated with Lactococcus lactis QMF 11, Listeria monocytogenes counts were maintained at the initial inoculum levels, not surpassing 2.3 log cfu mL-1 at the end of analysis. Regarding to Staphylococcus aureus, in the end of the experiment, cultures counts were 5.4 log cfu mL-1 (monocultures), 5.0 log cfu mL-1 (co-inoculation with Lactobacillus sakei) and 4.7 log cfu mL-1 (co-inoculation with Lactococcus lactis QMF 11). Despite the lower growth inhibition in comparison with Listeria monocytogenes, Staphylococcus aureus growth was significantly affect (p < 0.005) by the presence of Lactococcus lactis QMF 11, indicating that this strain has potential for use as biopreservative culture in dairy products.
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spelling Silva, Flávio Alves dahttp://lattes.cnpq.br/1713250447051507Alves, Virgínia Fariashttp://lattes.cnpq.br/5163169601493017Silva, Flávio Alves dahttp://lattes.cnpq.br/1713250447051507Campos, Maria Raquel HidalgoAndré, Maria Cláudia Dantas Porfirio BorgesSola , Marília CristinaDamiani , Clarissahttp://lattes.cnpq.br/0669403475574316Costa, Ana Carolina Cabral Carvalhaes2016-12-26T12:53:08Z2016-07-05COSTA, A. C. C. C. Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo. 2016. 58 f. Dissertação (Mestrado em Ciência e Tecnologia de Alimentos) - Universidade Federal de Goiás, Goiânia, 2016.http://repositorio.bc.ufg.br/tede/handle/tede/6628Due to their antagonistic activity against undesirable microorganisms, lactic acid bacteria (LAB) are a promising alternative for conservation and improvement of food safety. The aim of this work was isolate, from fresh Minas cheese, bacteriocin producing LAB with potential for use in biopreservation systems of dairy products. For this purpose, cheese samples were evaluated at the beginning and end of their shelf life and the isolation was performed by surface plating on MRS agar. The antimicrobial activity was verified by antagonism assay using Listeria monocytogenes ATCC 7644 and Staphylococcus aureus ATCC 25923 as indicators microorganisms. The selected LAB were submitted to the spot-on-the-lawn test, Gram stain, catalase and oxidase production and glucose metabolism in Hugh & Leifson broth. The occurrence of inhibition due to the production of organic acids and bacteriophages was discarded. The protein nature of antagonistic substances was confirmed by using proteases. After the well-diffusion assay, four LAB with satisfactory antagonistic activity against the indicators microorganisms were selected for identification by sequencing of the 16S RNA gene. All bacteria were identified as Lactococcus lactis. Based on antagonism assay, Lactococcus lactis QMF 11 was selected for use in co-inoculation studies with pathogens in pasteurized milk, kept at 8°C for 10 days. Lactobacillus sakei ATCC 15521 was used as negative control for bacteriocin production. After incubation period, monocultures of Listeria monocytogenes reached 8 log cfu mL-1 and in the presence of Lactobacillus sakei ATCC its population achieved 7.3 log cfu mL-1. However, when co-inoculated with Lactococcus lactis QMF 11, Listeria monocytogenes counts were maintained at the initial inoculum levels, not surpassing 2.3 log cfu mL-1 at the end of analysis. Regarding to Staphylococcus aureus, in the end of the experiment, cultures counts were 5.4 log cfu mL-1 (monocultures), 5.0 log cfu mL-1 (co-inoculation with Lactobacillus sakei) and 4.7 log cfu mL-1 (co-inoculation with Lactococcus lactis QMF 11). Despite the lower growth inhibition in comparison with Listeria monocytogenes, Staphylococcus aureus growth was significantly affect (p < 0.005) by the presence of Lactococcus lactis QMF 11, indicating that this strain has potential for use as biopreservative culture in dairy products.Devido à sua atividade antagonística contra micro-organismos indesejáveis, as bactérias ácido láticas (BAL) constituem uma alternativa promissora para a conservação e melhora da segurança de alimentos. O objetivo deste trabalho foi isolar, a partir de queijo Minas frescal, BAL produtoras de bacteriocinas com potencial para utilização em sistemas de bioconservação de produtos lácteos. Para tanto, amostras de queijo foram avaliadas no início e final de sua vida útil e o isolamento das BAL realizado por plaqueamento em ágar MRS. Verificou-se a atividade inibitória das bactérias pelo de ensaio de antagonismo em ágar usando Listeria monocytogenes ATCC 7644 e Staphylococcus aureus ATCC 25923 como micro-organismos indicadores. As BAL selecionadas nessa etapa foram submetidas aos testes spot-on-the-lawn, coloração de Gram, produção da catalase e oxidase e metabolismo da glicose em meio Hugh e Leifson. Descartou-se ocorrência de inibição pela produção de ácidos orgânicos e por bacteriófagos. A natureza proteica das substâncias antagonísticas foi confirmada utilizando-se proteases. Após realização do ensaio well diffusion, quatro BAL com atividade antagonística satisfatória frente aos micro-organismos indicadores utilizados foram selecionadas para identificação por sequenciamento do gene 16S do RNA. Todas foram identificadas como Lactococcus lactis. Com base nos testes de antagonismo, a cepa Lactococcus lactis QMF 11 foi selecionada para utilização no ensaio de co-inoculação com patógenos em leite pasteurizado mantido a 8oC por 10 dias. Lactobacillus sakei ATCC 15521 foi usado como controle negativo para produção de bacteriocinas. Após o período de incubação, Listeria monocytogenes em monocultura atingiu 8 log UFC/mL e na presença de Lactobacillus sakei chegou a 7,3 log UFC/mL. Entretanto, quando co-inoculada com Lactococcus lactis QMF11, as médias das contagens do patógeno ao final do experimento não ultrapassaram 2,3 log UFC/mL. Em relação a Staphylococcus aureus, as contagens finais foram: 5,4 log UFC/mL (monocultura), 5,0 log UFC/mL (co-inoculado com L. sakei) e 4,7 log UFC/mL (co-inoculado com Lactococcus lactis QMF 11). Apesar da menor inibição do crescimento de Staphylococcus aureus, em comparação à Listeria monocytogenes, sua multiplicação foi significantemente afetada (p<0.005) pela presença de Lactococcus lactis QMF 11, indicando que a cepa tem potencial para uso como cultura bioconservadora em produtos lácteos.Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2016-12-21T17:28:33Z No. of bitstreams: 2 Dissertação - Ana Carolina Cabral Carvalhaes Costa - 2016.pdf: 10341450 bytes, checksum: d69f0e1071d7458b683222711b4b5e4b (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-12-26T12:53:08Z (GMT) No. of bitstreams: 2 Dissertação - Ana Carolina Cabral Carvalhaes Costa - 2016.pdf: 10341450 bytes, checksum: d69f0e1071d7458b683222711b4b5e4b (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2016-12-26T12:53:08Z (GMT). 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dc.title.por.fl_str_mv Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo
dc.title.alternative.eng.fl_str_mv Isolation of bacteriocin - producing lactic acid bacteria and their activity against food pathogens in a biopreservation system of dairy products
title Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo
spellingShingle Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo
Costa, Ana Carolina Cabral Carvalhaes
Bactérias ácido láticas
Biopreservação
Lactococcus lactis
Listeria monocytogenes
Staphylococcus aureus
Lactic acid bacteria
Biopreservation
Lactococcus lactis
Listeria monocytogenes
Staphylococcus aureus
CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS
title_short Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo
title_full Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo
title_fullStr Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo
title_full_unstemmed Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo
title_sort Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo
author Costa, Ana Carolina Cabral Carvalhaes
author_facet Costa, Ana Carolina Cabral Carvalhaes
author_role author
dc.contributor.advisor1.fl_str_mv Silva, Flávio Alves da
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/1713250447051507
dc.contributor.advisor-co1.fl_str_mv Alves, Virgínia Farias
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/5163169601493017
dc.contributor.referee1.fl_str_mv Silva, Flávio Alves da
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/1713250447051507
dc.contributor.referee2.fl_str_mv Campos, Maria Raquel Hidalgo
dc.contributor.referee3.fl_str_mv André, Maria Cláudia Dantas Porfirio Borges
dc.contributor.referee4.fl_str_mv Sola , Marília Cristina
dc.contributor.referee5.fl_str_mv Damiani , Clarissa
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/0669403475574316
dc.contributor.author.fl_str_mv Costa, Ana Carolina Cabral Carvalhaes
contributor_str_mv Silva, Flávio Alves da
Alves, Virgínia Farias
Silva, Flávio Alves da
Campos, Maria Raquel Hidalgo
André, Maria Cláudia Dantas Porfirio Borges
Sola , Marília Cristina
Damiani , Clarissa
dc.subject.por.fl_str_mv Bactérias ácido láticas
Biopreservação
Lactococcus lactis
Listeria monocytogenes
Staphylococcus aureus
topic Bactérias ácido láticas
Biopreservação
Lactococcus lactis
Listeria monocytogenes
Staphylococcus aureus
Lactic acid bacteria
Biopreservation
Lactococcus lactis
Listeria monocytogenes
Staphylococcus aureus
CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS
dc.subject.eng.fl_str_mv Lactic acid bacteria
Biopreservation
Lactococcus lactis
Listeria monocytogenes
Staphylococcus aureus
dc.subject.cnpq.fl_str_mv CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS
description Due to their antagonistic activity against undesirable microorganisms, lactic acid bacteria (LAB) are a promising alternative for conservation and improvement of food safety. The aim of this work was isolate, from fresh Minas cheese, bacteriocin producing LAB with potential for use in biopreservation systems of dairy products. For this purpose, cheese samples were evaluated at the beginning and end of their shelf life and the isolation was performed by surface plating on MRS agar. The antimicrobial activity was verified by antagonism assay using Listeria monocytogenes ATCC 7644 and Staphylococcus aureus ATCC 25923 as indicators microorganisms. The selected LAB were submitted to the spot-on-the-lawn test, Gram stain, catalase and oxidase production and glucose metabolism in Hugh & Leifson broth. The occurrence of inhibition due to the production of organic acids and bacteriophages was discarded. The protein nature of antagonistic substances was confirmed by using proteases. After the well-diffusion assay, four LAB with satisfactory antagonistic activity against the indicators microorganisms were selected for identification by sequencing of the 16S RNA gene. All bacteria were identified as Lactococcus lactis. Based on antagonism assay, Lactococcus lactis QMF 11 was selected for use in co-inoculation studies with pathogens in pasteurized milk, kept at 8°C for 10 days. Lactobacillus sakei ATCC 15521 was used as negative control for bacteriocin production. After incubation period, monocultures of Listeria monocytogenes reached 8 log cfu mL-1 and in the presence of Lactobacillus sakei ATCC its population achieved 7.3 log cfu mL-1. However, when co-inoculated with Lactococcus lactis QMF 11, Listeria monocytogenes counts were maintained at the initial inoculum levels, not surpassing 2.3 log cfu mL-1 at the end of analysis. Regarding to Staphylococcus aureus, in the end of the experiment, cultures counts were 5.4 log cfu mL-1 (monocultures), 5.0 log cfu mL-1 (co-inoculation with Lactobacillus sakei) and 4.7 log cfu mL-1 (co-inoculation with Lactococcus lactis QMF 11). Despite the lower growth inhibition in comparison with Listeria monocytogenes, Staphylococcus aureus growth was significantly affect (p < 0.005) by the presence of Lactococcus lactis QMF 11, indicating that this strain has potential for use as biopreservative culture in dairy products.
publishDate 2016
dc.date.accessioned.fl_str_mv 2016-12-26T12:53:08Z
dc.date.issued.fl_str_mv 2016-07-05
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 COSTA, A. C. C. C. Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo. 2016. 58 f. Dissertação (Mestrado em Ciência e Tecnologia de Alimentos) - Universidade Federal de Goiás, Goiânia, 2016.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tede/6628
identifier_str_mv COSTA, A. C. C. C. Isolamento de bactérias láticas produtoras de bacteriocinas e avaliação de sua atividade frente a patógenos alimentares em sistema de bioconservação de produto lácteo. 2016. 58 f. Dissertação (Mestrado em Ciência e Tecnologia de Alimentos) - Universidade Federal de Goiás, Goiânia, 2016.
url http://repositorio.bc.ufg.br/tede/handle/tede/6628
dc.language.iso.fl_str_mv por
language por
dc.relation.program.fl_str_mv 4986785188683766460
dc.relation.confidence.fl_str_mv 600
600
600
600
dc.relation.department.fl_str_mv 4500684695727928426
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