Browsing by Subject "Lactic acid bacteria"

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Molekulare Interaktionen von Milchsäurebakterien mit enterohämorrhagischen Escherichia coli und humanen Darmepithelzellen
(2011) Stöber, Helen; Schmidt, Herbert
The interactions of 19 benign strains of lactic acid bacteria, bifidobacteria and staphylococci with five enterohemorrhagic Escherichia coli (EHEC) strains of different serotypes and virulence gene spectrum were investigated using a HT29 cell culture infection model. As a parameter for the infection the secretion of Interleukin 8 (IL-8) of the infected cells was analyzed by ELISA. None of the used benign strains induced an IL-8 secretion, whereas the infection with the EHEC strains leads ? independent of their virulence profile - to high amounts of IL-8. In coinfection assays with the pathogen EDL933 (O157:H7) and different test strains the secretion of IL-8 of the cultured cells was decreased by a few strains. With 12 of 19 tested strains, a weak reduction < 30 % of IL-8 secretion of HT29 cells after coinfection with EHEC O157:H7 strain EDL933 was observed. Six strains reduced the IL-8 secretion up to 60 % and the strain B. breve DSMZ 20083 decreased the IL-8 production about 73 %. Coinfection assays with different strains of one species (B. adolescentis DSMZ 20083 and DSMZ 20086 as well as L. johnsonii BFE 633 and DSMZ 10533) showed the strain specificity of the observed anti-inflammatory effect, due to different capabilities of IL-8 reduction. In further coinfection assays with different EHEC strains of the serotypes O103:H2, O26:H-, 0157:H- and O113:H21 different abilities of the benign strains to influence the infection with the different pathogen strains were noted. Therefore the protective anti-inflammatory effect is strain specific for the tested benign bacteria and also depends on the application of EHEC strains with different sero- and virulence types. Further investigations indicated the imperative of living bacteria for the observed protective effect; neither culture supernatant nor inactivated bacteria showed an effect on the IL-8 secretion of the EDL933 infected HT29 cells. The analysis of the cell culture supernatants 6 h after infection with different bacteria detected the production of lactic and acetic acid. The application of these acids in infection assays with EDL933 did not lead to an reduced IL-8 secretion of the infected cells. Therefore the production of organic acids did not explain the protective effect. The induction of IL-8 could not be traced back to the influence of a single virulence factor. Four PMK5 strains with deletions in different virulence genes induced similar IL-8 secretions in comparison to cells infected with the wild-type strain. Coinfection assays with the mutants and S. pasteuri LTH 5211 showed also similar IL-8 reductions than coinfection assays with the wild-type strain. It is to suppose that the anti-inflammatory effects of the benign bacteria do not influence a single virulence factor of the tested EHEC strains. As a second parameter the activation of the transcription factor ?Nuclear Factor kappa B? (NF-κB) of coinfected HT29 cells was monitored using a reporter-genassay. In comparison to the single EHEC-infection, the NF-κB activation was reduced by all tested lactic acid bacteria, bifidobacteria and S. pasteuri LTH 5211 in coinfection trials significantly. No strain-specificity and no pathogen-specificity could be observed. Interestingly, stimulation of the HT29 cells with benign bacteria led to inhibition of NF-κB activity, the measured values were less than the values of the negative control PBS. A gene expression analysis of toll-like receptors (TLRs), recognizing bacteria on cell surfaces and initiating the immune response, showed no regulation for TLR2. Infection with EDL933 led to down regulation of TLR4 and to up regulation of TLR9. Stimulation with L. rhamnosus GG, L. johnsonii DSMZ 10533 or L. fermentum DSMZ 20052 led neither to regulation of TLR4 nor TLR9. The benign bacteria did not influence the EHEC-induced TLR4 regulation in coinfection trials; in contrast the regulation of TLR9 was reduced significantly. The model described here is useful for screening basic effects of protective bacteria that are able to counteract EHEC-mediated effects on human cells and to study the molecular interaction between bacteria as well as between bacteria and human cultured cells.
Structure formation in fruit preparations by fruit fermentates produced with exopolysaccharide-forming lactic acid bacteria
(2025) Festini, Silvan; Zipori, Dor; Wallisch, Marc; Weiss, Agnes; Neidhart, Sybille; Schmidt, Herbert; Jekle, Mario
Fruit preparations are intermediate food products that are primarily used in the dairy industry for the production of fruit yogurt or frozen desserts. Typically, they are stabilized by added hydrocolloids like pectins. The objective of this study was to investigate the potential replacement of conventional stabilizers by structure-forming fermentates produced by exopolysaccharides (EPS)-forming lactic acid bacteria (LAB). Peach puree was selected as fermentation matrix. Prior to 72 h of incubation, it was inoculated with either the heterofermentative LAB strain Levilactobacillus brevis TMW 1.2112 or the homofermentative LAB strain Pediococcus parvulus strain LTH 1110, both being known to produce EPS in form of β-D-glucan. The lyophilized fermentates were applied as stabilizers to produce strawberry fruit preparations. Flow curves, viscoelastic behaviour and shear stability were measured to investigate the effect of fermentate incorporation on the rheological properties of the products. A fermentatively induced effect was observed in terms of a 1.3-fold increase in viscosity of strawberry model fruit preparations with 10 % fermentate of Lv. brevis TMW 1.2112 compared to the addition of the same dose of fermentate blank. Further, increasing the fermentate blank dose from 10 % to 15 % resulted in a 2.4-fold viscosity increase of the model fruit preparations. High shear stability was found in all model strawberry fruit preparations. However, fermentation had no clear benefit in terms of viscoelastic behaviour and shear stability of the fruit preparations. Although the fermentatively induced thickening potential was limited, production of viscosity-increasing peach fermentate with minor changes in the sugar and amino acid profiles of the fruit proved to be feasible.