Mean TER values did not differ after 1-3 h of incubation (P > 0.05), but significantly decreased after 24 h of incubation (Figure 3). In contrast, TER measured for pure cultures of S. Typhimurium N-15 in buffered DMEM showed a continuous and pronounced decrease in TER (Figure 3). Compared to initial model stabilization periods (Stab),
mean TER measured 1-3 h after incubation with effluents of all reactors from Salmonella infection periods (Sal) AUY-922 research buy were significantly lower (P < 0.0001, Table 1), with a mean decrease of 40 ± 4% (Figure 2D). This effect on cell integrity was confirmed by confocal microscopy analysis which demonstrated highly disrupted tight junctions after Salmonella infection for distal reactor (R3) effluents of F1 (Figure 4B) compared to initial
model stabilization periods (Figure 4A). E. coli L1000 stimulates Salmonella growth yet reduces invasion in the distal colon region E. coli L1000 established itself in the three-stage model at low levels with slightly but non-significantly higher numbers measured in R3 (4.9 ± 0.9 log10 MCN/ml) compared to R1 (4.5 ± 0.6 Midostaurin purchase log10 MCN/ml) and R2 (4.3 ± 0.6 log10 MCN/ml; Figure 2A). As shown previously [15], the addition of E. coli L1000 beads to the intestinal fermentation model enhanced Salmonella growth in all colon reactors compared to initial Salmonella infection periods (Sal; Figure 2A). However, significantly lower Salmonella invasion ratios were measured many with transverse and distal reactor effluents (Figure 2B) in comparison with initial Salmonella stabilization periods (Sal). Concomitantly, Salmonella adhesion ratios remained stable in R3 (Figure 2B), however the efficiency of cell-associated Salmonella to invade HT29-MTX
cells (Figure 2C) decreased significantly. The second addition of E. coli L1000 (Ecol II) had no further effects on Salmonella adhesion and invasion ratios in R1 and R3. However, a significantly enhanced (P = 0.0004) Salmonella invasion ratio was measured with transverse reactor effluents (Figure 2B) compared to the first E. coli L1000 period (Ecol I), which was accompanied by a significant increase in invasion efficiency (Figure 2C). Similar mean TER values were measured with effluents from first E. coli L1000 (Ecol I) and Salmonella colonization (Sal) periods for all reactors (Table 1, Figure 2D), despite significantly higher Salmonella counts (P < 0.01) after the addition of E. coli L1000 (Figure 2A). TER significantly (P > 0.05) decreased by 19% and 26% with transverse and distal reactor effluents respectively (Figure 2D) after the second addition of E. coli L1000 (Ecol II) compared to the previous period (Ecol I) while Salmonella counts did not change for the two E. coli periods (Figure 2A). B. thermophilum RBL67 exerts a protective effect on epithelial integrity in highly infected environments B.
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