Typhimurium N-15 in presence of a complex intestinal microbiota and to assess the host-protection properties of E. coli L1000 and B. thermophilum RBL67 sequentially inoculated in the infection model, as well as the protective effect of inulin. Effluent samples were produced in two three-stage
continuous colonic models, mimicking the proximal, transverse and distal colon regions and inoculated with immobilized child fecal microbiota and Salmonella, and used to test the effects of probiotics and inulin on gut microbiota composition and metabolism, and on Salmonella growth [15]. Effluents collected from different fermentation periods were directly applied to HT29-MTX cells to measure Salmonella invasion and monitor changes in cellular integrity through both measurement of transepithelial electrical resistance (TER) and confocal microscopy. Data from VS-4718 complex effluents were compared with pure Salmonella cultures. Results Complex reactor effluents were collected during pseudo-steady states (last 3 days) of different experimental periods from two continuous three-stage colonic fermentation models as indicated in GDC-0994 Figure 1 and applied directly onto confluent mucus-secreting
HT29-MTX cells. Temporal and environmental factors affecting bacterial growth, Salmonella invasion and TER across cell monolayers Akt inhibitor are summarized in Figure 2 and Table 1. TER across cell monolayers after incubation with simple and complex fermentation samples are compared in Figure 3 and the effects on epithelial integrity upon effluent application are shown in Figure 4. Figure 1 Experimental design of continuous three-stage colonic fermentations.
Two three-stage continuous fermentation models (F1 and F2) simulating (R1) proximal, (R2) transverse and (R3) distal colonic sections were inoculated with the same immobilized child fecal microbiota, infected with Salmonella beads and operated in parallel for a total of 65 days divided into different experimental periods as described previously [15]. For this study, reactor effluents collected Gemcitabine molecular weight during the last 3 days of each experimental period were directly applied onto confluent mucus-secreting HT29-MTX cell layers to detect host-protection properties of different experimental treatments. Data obtained during similar treatments in models F1 and F2 (highlighted in the same color) were not significantly different and therefore used as repetitions: (Stab) initial system stabilization periods, (Sal) Salmonella infection periods, (Ecol) E. coli L1000 wt treatments (microcin B17-producing wild-type strain), (Ecol*) E. coli L1000 MccB17- treatments (microcin B17-negative mutant strain), (Bif) B. thermophilum RBL67 treatments, (Inulin) prebiotic inulin treatment. Figure 2 Bacterial growth, Salmonella invasion and TER across HT29-MTX monolayers are affected by experimental and environmental factors.