Orientadores: Gláucia Maria Pastore, Rosângela dos Santos

Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos

Resumo: Estudos sao realizados com oligossacarideos, probioticos e a combinação deles; sao varios os fatores que envolvem os beneficios que estes em combinacao podem agregar para a saude dos consumidores. Esta pesquisa teve como objetivo avaliar o comportamento simbiotico in vitro das linhagens...

Resumo: Estudos sao realizados com oligossacarideos, probioticos e a combinação deles; sao varios os fatores que envolvem os beneficios que estes em combinacao podem agregar para a saude dos consumidores. Esta pesquisa teve como objetivo avaliar o comportamento simbiotico in vitro das linhagens probioticas Bifidobacterium animalis (Bb-12) e Lactobacillus acidophillus (LA-05), em substratos enriquecidos com oligossacarideos, ou seja, avaliar se esses potencializavam a capacidade probiotica das linhagens estudadas. Os oligossacarideos utilizados no estudo foram o galactooligossacarideo (GOS) sintetizado pela ?-galactosidase de Scopulariops sp., frutooligossacarideo (FOSOrafti) e extrato bruto de Yacon. Avaliou-se o efeito bifidogenico de diferentes concentracoes de GOS, FOS e Yacon utilizando as culturas probioticas e sua capacidade de acidificacao do meio. Foi avaliado o perfil hidrofobico e acido da membrana celular dos probioticos usando meio enriquecido com GOS, FOS e controle meio MRS (Man Rogosa e Sharpe) sem fonte de dextrose; pelo método MATH (aderencia microbiana de hidrocarbonetos). Realizou-se tambem a producao, extracao e quantificacao de exopolissacarideos (EPS) por Lactobacillus acidophillus (LA-05) em meios enriquecidos com FOS e GOS, a quantificacao foi pelo metodo fenol-sulfurico seguida de leitura de absorbancia a 490nm, os resultados foram submetidos a uma curva padrao de glicose. Verificou-se ainda o antagonismo das linhagens probioticas cultivadas em meios suplementados com GOS, FOS, Yacon, MRS sem fonte de dextrose e controle MRS adicionado de NaOH, sobre linhagens patogenicas; o antagonismo foi avaliado atraves do metodo de difusao em agar. Avaliou tambem a capacidade de producao de acido latico. Na avaliacao bifidogenica os resultados revelaram que os substratos estimularam o metabolismo dos probioticos estudados. A melhor atividade para as linhagem de Lactobacillus acidophillus (LA-05) foi com o substrato GOS com atividade de 11,56 LogUFC (na concentracao de 200mg de GOS, em 12 horas de incubacao), e para e Bifidobacterium animalis (Bb12) foi com o substrato FOS com atividade de 11,3 LogUFC (na concentracao de 100mg FOS, em 12 horas de incubacao), na medida que se aumentou a concentracao dos substratos, houve a reducao da atividade bifidogenica. Os resultados revelaram que os prebioticos GOS e FOS estimularam o perfil de adesao e acidez da membrana celular das linhagens probioticas estudadas, quando comparado com o controle meio MRS sem fonte de dextrose. Quanto a producao de EPS, o maior valor (13,53 \g.mL) foi obtido apos 48 horas de cultivo em caldo MRS suplementado com GOS (25% v/v), enquanto que o FOS (25% v/v) tambem estimulou a producao (7,68 \g.mL) em 12 horas, seguida da queda de producao apos este periodo. Verificou-se que os oligossacarideos estimularam a acao antagonica dos probioticos sobre os micro-organismos patogenicos, e que esta inibicao do crescimento pode estar ligada com a producao de acidos pelos probioticos em conjunto com os oligossacarideos. Em sintese, os resultados revelaram que os oligossacarideos estimularam o potencial probiotico das linhagens estudadas

Abstract: Several studies have been conducted with oligosaccharides in general and also with probiotics, or combination of them, due to factors involving the combined benefits they can add to the health of consumers. This research aimed to evaluate in vitro the behavior of symbiotic probiotic...

Abstract: Several studies have been conducted with oligosaccharides in general and also with probiotics, or combination of them, due to factors involving the combined benefits they can add to the health of consumers. This research aimed to evaluate in vitro the behavior of symbiotic probiotic strains Bifidobacterium animalis (Bb-12) and Lactobacillus acidophilus (LA-05), on substrates enriched with oligosaccharides in the case to assess whether the oligosaccharides intensified the ability of probiotic strains studied. The oligosaccharides used in this study were synthesized by the galactooligosaccharide ?-galactosidase Scopulariops sp., fructooligosaccharide and crude extract of yacon. We evaluated the bifidogenic effect of different concentrations of GOS, FOS and Yacon using probiotic cultures and their ability to acidification of the medium. Was evaluated the profile acid and hydrophobic cellular membrane of probiotics using medium supplemented with GOS, FOS and control MRS medium without added dextrose; MATH method (microbial adherence to hydrocarbons). Was also conducted production, extraction and quantification of exopolysaccharides (EPS) by Lactobacillus acidophilus (LA-05) media supplemented with FOS and GOS, quantification was by the phenolsulfuric method followed by absorbance reading at 490nm, the results were subjected to a standard curve of glucose. It was further antagonism of probiotic strains grown in media supplemented with GOS, FOS, yacon, MRS without dextrose and source control MRS added NaOH on pathogenic strains; antagonism was assessed by the agar diffusion method; also evaluated the ability to produce lactic acid. In evaluating the results revealed that bifidogenic substrates stimulated metabolism studied probiotics, the best activity for the strain of Lactobacillus acidophilus (LA-05) was the substrate with GOS activity of 11.56 LogUFC (200mg in 12 hours incubation), and for and Bifidobacterium animalis (Bb-12) was with the FOS substrate with activity of 11.3 LogUFC (100mg in 12 hours incubation), as it increased the concentration of the substrates decreased the activity of microorganisms. The results revealed that prebiotic GOS and FOS stimulated acid profile of adhesion and cell membrane of probiotic strains studied, as compared to the MRS control medium without added dextrose. As for the highest production of EPS production (13.53 ?g.ml) was after 48 hours of culture in MRS broth supplemented with GOS, while also stimulated FOS production (7.68 ?g.ml) for 12 hours followed by production decrease thereafter. It was found that the oligosaccharides stimulated antagonistic action of probiotics on pathogenic microorganisms and that this growth inhibition is connected with acid production by probiotics together with the oligosaccharides. In summary, the results revealed that the oligosaccharides stimulated the potential probiotic strains studied