Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes

Detalhes bibliográficos
Autor(a) principal: Fernandes, Mariane Silveira Magalhães
Data de Publicação: 2019
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
dARK ID: ark:/83112/0013000024bfk
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/40355
Resumo: The term biofilm describes the life form characterized by the microbial adhesion to a surface with the production of extracellular polymeric substances, constituting a gelatinous network that protects the cells. Biofilms formation is involved in the increasing incidence of important multidrug resistant bacterial pathogens to conventional antimicrobial therapies, being verified the presence of enterobacteria isolated from fresh foods resistant to multiple antibiotics. Given this context, it is necessary to search for new forms of microbial control. Lactic acid bacteria, including Lactobacillus species, have been associated with the prevention or dispersion of biofilms formed by pathogenic microorganisms. Therefore, the aim of this work was to evaluate the antimicrobial and antibiofilm activity of the cell-free supernatants produced by Lactobacillus strains on multi-drug resistant E. coli isolates. For this, the profile of antimicrobial susceptibility of E. coli strains was identified by the VITEK®2 automated system. For the evaluation of the antagonistic activity of probiotics, the Spot Overlay technique was used, with measurement of the inhibition halos. The cell-free supernatant produced by Lactobacillus was tested for its antimicrobial effect by plaque microdilution of the supernatant with the concentrations ranging 6,25 a 50%. The antibiofilm action was evaluated after treatment of preformed biofilm for 12 h, using a Crystal Violet staining technique to verify changes in biofilm biomass. As a result, the samples were found to be resistant to ampicillins, cephalothin and quinolones. As for the antagonism test, all probiotic strains demonstrated high inhibitory capacity (inhibition scores > 6 mm) against multiresistant E. coli isolates. The isolates were susceptible to antimicrobial activity of the supernatants produced, with Minimal Inhibitory and Bactericidal Concentrations ranging from 12.5 to 50%. However, no effect was observed after neutralization, which indicate that the inhibitory action may be related to the production of organic acids. Regarding the antibiofilm effect, all the analyzed probiotics were capable to reduce the preformed biofilm, standing out the commercial strain, that was capable to inhibit around 50% of the biomass. Therefore, the cellfree preparations produced by Lactobacillus from the present study may represent tools for a possible overcome of resistance mechanisms presented by E. coli isolates.
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spelling Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentesAntimicrobial activity and antibiofilm of the supernatant of lactobacillus cell-free strains on pharmacoresistant escherichia coli isolatesEnterobactériasProbióticosAntimicrobianosThe term biofilm describes the life form characterized by the microbial adhesion to a surface with the production of extracellular polymeric substances, constituting a gelatinous network that protects the cells. Biofilms formation is involved in the increasing incidence of important multidrug resistant bacterial pathogens to conventional antimicrobial therapies, being verified the presence of enterobacteria isolated from fresh foods resistant to multiple antibiotics. Given this context, it is necessary to search for new forms of microbial control. Lactic acid bacteria, including Lactobacillus species, have been associated with the prevention or dispersion of biofilms formed by pathogenic microorganisms. Therefore, the aim of this work was to evaluate the antimicrobial and antibiofilm activity of the cell-free supernatants produced by Lactobacillus strains on multi-drug resistant E. coli isolates. For this, the profile of antimicrobial susceptibility of E. coli strains was identified by the VITEK®2 automated system. For the evaluation of the antagonistic activity of probiotics, the Spot Overlay technique was used, with measurement of the inhibition halos. The cell-free supernatant produced by Lactobacillus was tested for its antimicrobial effect by plaque microdilution of the supernatant with the concentrations ranging 6,25 a 50%. The antibiofilm action was evaluated after treatment of preformed biofilm for 12 h, using a Crystal Violet staining technique to verify changes in biofilm biomass. As a result, the samples were found to be resistant to ampicillins, cephalothin and quinolones. As for the antagonism test, all probiotic strains demonstrated high inhibitory capacity (inhibition scores > 6 mm) against multiresistant E. coli isolates. The isolates were susceptible to antimicrobial activity of the supernatants produced, with Minimal Inhibitory and Bactericidal Concentrations ranging from 12.5 to 50%. However, no effect was observed after neutralization, which indicate that the inhibitory action may be related to the production of organic acids. Regarding the antibiofilm effect, all the analyzed probiotics were capable to reduce the preformed biofilm, standing out the commercial strain, that was capable to inhibit around 50% of the biomass. Therefore, the cellfree preparations produced by Lactobacillus from the present study may represent tools for a possible overcome of resistance mechanisms presented by E. coli isolates.A formação de biofilmes está envolvida com o aumento da incidência de importantes patógenos bacterianos multirresistentes à terapias antimicrobianas convencionais, sendo constatada a presença de enterobactérias resistentes a múltiplos antibióticos em alimentos in natura. Dessa forma, surge a necessidade da busca por novas formas de controle microbiano. Como possível estratégia de prevenção, espécies de Lactobacillus vêm sendo associadas à dispersão de biofilmes formados por micro-organismos patogênicos. Portanto, o objetivo deste trabalho foi analisar a atividade antimicrobiana e antibiofilme de sobrenadantes de Lactobacillus cell-free sobre isolados de Escherichia coli multirresistentes. Para isto, foram utilizadas quatro cepas de E. coli isoladas de filés de tilápia (P12, P25, P35 e P36) e três cepas probióticas (LA 14, L. acidophilus ATCC 4356 e L. rhamnosus ATCC 9595). O perfil de susceptibilidade antimicrobiana dos isolados de E. coli foi verificado por meio de sistema automatizado VITEK ® 2. Para avaliação da atividade antagônica dos probióticos foi utilizada a técnica de spot overlay, com medição do diâmetro de inibição. Os sobrenadantes cell-free produzidos foram testados quanto ao seu efeito antimicrobiano, por meio de microdiluição em placa entre as concentrações de 6,25 a 50%. A ação antibiofilme foi avaliada após tratamento de biofilme pré-formado por 12 h pelas cepas de E. coli, por meio da técnica de coloração por cristal violeta a fim de verificar alterações sobre a biomassa de biofilme. Como resultado, verificou-se que as amostras apresentam um perfil de resistência a ampicilinas, cefalotina e quinolonas. Quanto ao teste de antagonismo, todas as cepas probióticas demonstraram alta capacidade de inibição (escore de inibição > 6 mm) sobre os isolados de E. coli multirresistentes. As cepas patogênicas foram susceptíveis a atividade antimicrobiana dos sobrenadantes produzidos, com Concentração Inibitória e Bactericida Mínima variarando entre 12,5 a 50%. Entretanto, os mesmos não mantiveram seu efeito após neutralização, o que pode sinalizar que o resultado inibitório esteja relacionado com a produção de ácidos orgânicos. Com relação ao efeito antibiofilme, todos os probióticos analisados foram capazes de reduzir o biofilme pré-formado, com destaque para a cepa comercial que apresentou percentuais de inibição por volta de 50% da biomassa. Portanto, as preparações cell-free produzidas pelos Lactobacillus do presente estudo podem representar ferramentas para uma possível superação dos mecanismos de resistência apresentados por isolados E. coli.Carneiro, Victor AlvesFernandes, Mariane Silveira Magalhães2019-03-20T18:13:48Z2019-03-20T18:13:48Z2019-02-25info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfFERNANDES, M.S.M. Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes. 2019 63 f. Dissertação (Mestrado em Biotecnologia) - Campus de Sobral, Universidade Federal do Ceará, Sobral, 2019.http://www.repositorio.ufc.br/handle/riufc/40355ark:/83112/0013000024bfkporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2019-03-20T18:13:48Zoai:repositorio.ufc.br:riufc/40355Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:59:54.110868Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes
Antimicrobial activity and antibiofilm of the supernatant of lactobacillus cell-free strains on pharmacoresistant escherichia coli isolates
title Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes
spellingShingle Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes
Fernandes, Mariane Silveira Magalhães
Enterobactérias
Probióticos
Antimicrobianos
title_short Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes
title_full Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes
title_fullStr Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes
title_full_unstemmed Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes
title_sort Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes
author Fernandes, Mariane Silveira Magalhães
author_facet Fernandes, Mariane Silveira Magalhães
author_role author
dc.contributor.none.fl_str_mv Carneiro, Victor Alves
dc.contributor.author.fl_str_mv Fernandes, Mariane Silveira Magalhães
dc.subject.por.fl_str_mv Enterobactérias
Probióticos
Antimicrobianos
topic Enterobactérias
Probióticos
Antimicrobianos
description The term biofilm describes the life form characterized by the microbial adhesion to a surface with the production of extracellular polymeric substances, constituting a gelatinous network that protects the cells. Biofilms formation is involved in the increasing incidence of important multidrug resistant bacterial pathogens to conventional antimicrobial therapies, being verified the presence of enterobacteria isolated from fresh foods resistant to multiple antibiotics. Given this context, it is necessary to search for new forms of microbial control. Lactic acid bacteria, including Lactobacillus species, have been associated with the prevention or dispersion of biofilms formed by pathogenic microorganisms. Therefore, the aim of this work was to evaluate the antimicrobial and antibiofilm activity of the cell-free supernatants produced by Lactobacillus strains on multi-drug resistant E. coli isolates. For this, the profile of antimicrobial susceptibility of E. coli strains was identified by the VITEK®2 automated system. For the evaluation of the antagonistic activity of probiotics, the Spot Overlay technique was used, with measurement of the inhibition halos. The cell-free supernatant produced by Lactobacillus was tested for its antimicrobial effect by plaque microdilution of the supernatant with the concentrations ranging 6,25 a 50%. The antibiofilm action was evaluated after treatment of preformed biofilm for 12 h, using a Crystal Violet staining technique to verify changes in biofilm biomass. As a result, the samples were found to be resistant to ampicillins, cephalothin and quinolones. As for the antagonism test, all probiotic strains demonstrated high inhibitory capacity (inhibition scores > 6 mm) against multiresistant E. coli isolates. The isolates were susceptible to antimicrobial activity of the supernatants produced, with Minimal Inhibitory and Bactericidal Concentrations ranging from 12.5 to 50%. However, no effect was observed after neutralization, which indicate that the inhibitory action may be related to the production of organic acids. Regarding the antibiofilm effect, all the analyzed probiotics were capable to reduce the preformed biofilm, standing out the commercial strain, that was capable to inhibit around 50% of the biomass. Therefore, the cellfree preparations produced by Lactobacillus from the present study may represent tools for a possible overcome of resistance mechanisms presented by E. coli isolates.
publishDate 2019
dc.date.none.fl_str_mv 2019-03-20T18:13:48Z
2019-03-20T18:13:48Z
2019-02-25
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.uri.fl_str_mv FERNANDES, M.S.M. Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes. 2019 63 f. Dissertação (Mestrado em Biotecnologia) - Campus de Sobral, Universidade Federal do Ceará, Sobral, 2019.
http://www.repositorio.ufc.br/handle/riufc/40355
dc.identifier.dark.fl_str_mv ark:/83112/0013000024bfk
identifier_str_mv FERNANDES, M.S.M. Atividade antimicrobiana e antibiofilme do sobrenadante de cepas de lactobacillus cell-free sobre isolados de escherichia coli farmacoresistentes. 2019 63 f. Dissertação (Mestrado em Biotecnologia) - Campus de Sobral, Universidade Federal do Ceará, Sobral, 2019.
ark:/83112/0013000024bfk
url http://www.repositorio.ufc.br/handle/riufc/40355
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
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institution UFC
reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
repository.mail.fl_str_mv bu@ufc.br || repositorio@ufc.br
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