Fluctuations in butyrate-producing bacteria in ulcerative colitis patients of North India

Figure 1 Flow cytometric analysis of fecal microflora using 16S rRNA targeted probes. A: The region R1 corresponding to relative size (the forward angle light scatter)/granularity (the side angle light scatter) of the bacteria was delineated. This region R1 was gated for further dot plots; B: Bacteria from fecal samples were hybridized with NONEUB338 probe; C: EUB338 and Erec 482; D: EUB338 and Clep 1156. A shift in fluorescence to higher intensities was obtained upon hybridization with positive control or group-specific probe: right lower and upper quadrant compared to the lower left quadrant. The signal in lower left quadrant represents debris. The events in upper right quadrant represent the proportion of bacterial cells hybridized with the group-specific probe within the total bacterial cells hybridized with the universal bacterial probe EUB338-fluorescein isothiocyanate (FITC). The enumeration of targeted cells was corrected by subtracting the background fluorescence, which was measured using the negative control NONEUB338 probe. Fluorescent in situ hybridization (FISH)-flow cytometry data were expressed as the mean % group-specific probe/EUB338 ± SE as enumerated by FISH-flow cytometry in control, moderate, severe and remission samples of ulcerative colitis; E: Erec 482; F: Clep 1156.

Figure 2 Quantitative polymerase chain reaction data showing the numbers of bacteria (± SE) in fecal samples of control vs ulcerative colitis patients. A: Clostridium coccoides; B: Clostridium leptum. The Y axis represents number of bacteria and X axis represents the sample category.

Figure 3 Short chain fatty acids in fecal samples of ulcerative colitis patients vs control samples, analyzed by gas chromatography. A: Acetate; B: Iso-butyrate; C: Butyrate. The Y axis represents concentration of respective short chain fatty acid and X axis represents the sample category.

Figure 4 Analysis of predominant butyrate-producing bacteria. A-C: Fecalibacterium prausnitzii (A), Roseburia intestinalis (B), and Eubacterium rectale (C) by fluorescent in situ hybridization-flow cytometry; D-F: Fecalibacterium prausnitzii (D), Roseburia intestinalis (E), and Eubacterium hallii (F) by quantitative polymerase chain reaction. The Y axis represents number of bacteria and X axis represents the status of patients. Horizontal bars with asterisks represent comparison between ulcerative colitis and control conditions.