Role of Intestinal Microbiota in Ulcerative Colitis

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Introduction and objectives Introduction and objectives Since the beginning of the last century from the early work of Elie Metchnikoff, it is believed that a synergistic interaction exists between intestinal microbes and their host. Metchnikoff observed that: “… different susceptibilities of people to the harmful action of microbes and their products. Some can swallow without any evil result a quantity of microbes which in the case of other individuals would produce a fatal attack of cholera. Everything depends upon the resistance offered to the microbes by the invaded organism" (Metchnikoff, 1908). Hence, an unfavorable composition of the gut microbiota could play a role in intestinal infections and diseases. Consumption of indigestible carbohydrates or dietary fibers is suggested to improve gut health by modulating the gut environment through selective stimulation of bacteria who are able to promote beneficial events in the host by inhibition of pathogenic bacteria, and induction of immune regulatory functions (Vernazza et al., 2006). The objectives of this thesis were to investigate, if the gut microbiota and its ability to colonize mucus are different in patients suffering from the intestinal bowel disease, ulcerative colitis (UC) than in healthy subjects, hence playing a role in colonic inflammation. Additionally, investigate the ability of novel indigestible carbohydrates or fibers from sugar beet and potato pulp to induce beneficial changes in fecal microbiota communities in vitro obtained from human volunteers both healthy and UC patients. The first part of this thesis gives a theoretical introduction to the phylogenetic composition of the human intestinal microbiota, the location of the microbiota within the colon and their interactions with the host. Additionally, UC and the colonic microbiota composition observed in UC patients will be described. Finally, modulation of the intestinal microbiota by prebiotics will be addressed together with a description of the production of prebiotics and novel carbohydrates. The second part contains a description of the methodologies used in the studies of this thesis and six papers that demonstrate the experimental work. The last part is discussion and conclusion based on the theoretical and experimental work of this thesis. X
List of manuscripts Paper 1 List of Manuscripts Vigsnæs, L.K., Brynskov, J., Wilcks, A., and Licht, T.R. Gram‐negative bacteria account for differences in fecal microbiota between patients with ulcerative colitis and healthy controls. (2011) Submitted to BMC Microbiology, under review. Paper 2 Vigsnaes, L.K., Van den Abbeele, P., Van de Wiele, T. and Licht, T.R. Fecal lactobacilli, and bifidobacteria from ulcerative colitis patients display reduced ability to colonize mucus in the M‐ SHIME. (2011) Paper in preparation. Paper 3 Vigsnæs, L.K., Holck, J., Meyer, A.S, and Licht, T.R. (2011) In vitro Fermentation of Sugar Beet Arabino‐Oligosaccharides by Fecal Microbiota Obtained from Patients with Ulcerative Colitis Selectively Stimulates the Growth of Bifidobacterium spp. and Lactobacillus spp. Applied and Environmental Microbiology. 77:8336‐8344 Paper 4 Holck, J., Lorentzen, A., Vigsnæs, L.K., Licht, T.R, Mikkelsen, J.D. and Meyer, A.S. (2011) Feruloylated and non‐feruloylated arabino‐oligosaccharides from sugar beet pectin selectively stimulates the growth of Bifidobacterium spp. in human fecal in vitro fermentation. Journal of Agricultural and Food Chemistry. 59:6511‐6519. Paper 5 Thomassen, L.V., Vigsnæs, L.K., Licht, T.R., Mikkelsen, J.D., and Meyer, A.S. (2011) Maximal release of highly bifidogenic soluble dietary fibers from industrial potato pulp by minimal enzymatic treatment. Applied Microbiology and Biotechnology. 90:873‐884. Paper 6 Holck, J., Hjernø, K., Lorentzen, A., Vigsnæs, L.K., Hemmingsen.L., Licht, T.R., Mikkelsen, J.D., Meyer, A.S. (2011) Tailored enzymatic production of oligosaccharides from sugar beet pectin and evidence of differential effects of a single DP chain length difference human fecal microbiota composition after in vitro fermentation. Process Biochemistry. 46 :1039‐1049. XI
Page 1: Role of Intestinal Microbiota in Ul
Page 4 and 5: Role of Intestinal Microbiota in Ul
Page 6 and 7: Preface Preface This thesis present
Page 8 and 9: Summary Summary The microbiota of t
Page 10 and 11: Dansk sammendrag Dansk sammendrag M
Page 14 and 15: List of Manuscripts Not included in
Page 16 and 17: List of contents List of Centents P
Page 18 and 19: List of Centents Methodology append
Page 21 and 22: 1. The intestinal environment Theor
Page 23 and 24: Theoretical part 5 1. The intestina
Page 25 and 26: 2. The colonic environment Theoreti
Page 27 and 28: Theoretical part 9 2. The colonic e
Page 29 and 30: Table 1: The presence of glycoside
Page 31 and 32: Theoretical part Figure 3: The colo
Page 33 and 34: 3. Inflammatory Bowel disease Theor
Page 35 and 36: Theoretical part 17 3. Inflammatory
Page 37 and 38: Theoretical part 19 4. Modulation o
Page 43 and 44: Table 4: Clinical trials on the pre
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Abstract Background Detailed knowle
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depending the level of disease acti
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in 1 x TAE at 60 °C for 16 h at 36
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Statistics PCA were generated by SA
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The PCA of the Gram‐positive bact
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layer of UC patients and found that
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Acknowledgements The authors thank
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Table 2 ‐ 16S rRNA gene and 16S
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1. Firmicutes phylum 2. Bacteroidet
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Supplementary Figure S1. Dice clust
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Reference List 1. Ahmed S, Macfarla
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32. Matsuki T, Watanabe K, Fujimoto
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Methodology part Paper 2 Fecal lact
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Fecal lactobacilli and bifidobacter
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Introduction The mucus layer lining
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efore enrolment and there was no si
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(Bio‐Rad Labs, Hercules, Californ
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Microbial community analysis using
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difference from the luminal microbi
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that C. coccoides group and C. lept
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Table 1 ‐ 16S rRNA gene of phylum
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Table 2 ‐ Preference of bacterial
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Figure 1. A) Schematic overview of
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A. B. Figure 3. Principal component
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15. Fooks LJ, Gibson GR. (2002) In
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47. Ouwehand AC, Suomalainen T, Tol
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Methodology part Paper 3 Paper 3 In
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APPLIED AND ENVIRONMENTAL MICROBIOL
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8338 VIGSNÆS ET AL. APPL. ENVIRON.
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Methodology part Introduction The a
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Journal of Agricultural and Food Ch
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Methodology part Paper 5 Paper 5 Ma
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Appl Microbiol Biotechnol (2011) 90
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Methodology part Paper 6 Tailored e
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Process Biochemistry 46 (2011) 1039
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Table 1 List of enzymes. Enzyme Sou
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J. Holck et al. / Process Biochemis
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Intensity 50 0 C1 175.05 J. Holck e
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J. Holck et al. / Process Biochemis
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[29] Bauer S, Vasu P, Persson S, Mo
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Methodology part 150
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Methodology part 152 Appendix 1 hor
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Appendix 3 Methodology part Appendi
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Methodology part 156 Appendix 3
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7. Discussion and perspectives Disc
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Discussion and conclusion 161 7. Di
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References References Abreu,M.T., V
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References Derrien,M., Vaughan,E.E.
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References Haarman,M. and Knol,J. (
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References Lantz,P.G., Matsson,M.,
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References Matsuki,T., Watanabe,K.,
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References Pullan,R.D., Thomas,G.A.
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References Tannock,G.W. (2010). Ana
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National Food Institute Technical U
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Role of Intestinal Microbiota in Ulcerative Colitis

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