Figure 4 Lack of protective effects of dietary supplementation with non-aggregating mutant L. crispatus MU5 on DSS colitis. DSS colitis was induced as described in Section 2, 24 h after the induction of colitis animals were randomly divided to receive once a day (for 3 days) either 10⁸L. crispatus M247, 10⁸L. crispatus MU5 in MRS medium or 10⁸L. crispatus MU5 in a 30% sucrose solution. Severity of colitis was determined by body weight change (panel a), results are shown as percent change over the initial body weight, and MPO activity in full thickness sections of proximal colon, expressed as U/mg wet tissue (panel b). As shown in panels a and b administration of non-aggregating mutant L. crispatus MU5 resulted in the complete loss of the protective effects, whereas administration of the adhesion deficient strain in a 30% sucrose solution restore the protective effect on colitis. Values are means ± SEM. ^*P < 0.05 vs control; ^**P < 0.01 vs control, ⁺P < 0.05 vs DSS alone; ⁺⁺P < 0.01 vs DSS alone. From: Beneficial effect of auto-aggregating Lactobacillus crispatus on experimentally induced colitis in mice FEMS Immunol Med Microbiol. 2005;43(2):197-204. doi:10.1016/j.femsim.2004.08.011 FEMS Immunol Med Microbiol | © 2004 Federation of European Microbiological Societies.

Figure 3 Effect of conditioning the colonic lumen with L Figure 3 Effect of conditioning the colonic lumen with L. crispatus M247 before the onset of DSS colitis. L. crispatus M247 (10⁸ bacteria) were administered daily for 5 consecutive days and then stopped at the time of DSS administration. Colitis induced body weight loss and colonic mucosal MPO levels were determined after 5 days. Panel a shows the absence of any protective effect of pre-conditioning of colonic lumen by L. crispatus M247 on DSS-induced weight loss. Panel b shows that pre-conditioning of colonic lumen with L. crispatus M247 reduced by 30% mucosal MPO activity (P < 0.05) as compared to DSS treated mice. Values are means ± SEM. ^*P < 0.05 vs control; ^**P < 0.01 vs control. From: Beneficial effect of auto-aggregating Lactobacillus crispatus on experimentally induced colitis in mice FEMS Immunol Med Microbiol. 2005;43(2):197-204. doi:10.1016/j.femsim.2004.08.011 FEMS Immunol Med Microbiol | © 2004 Federation of European Microbiological Societies.

Figure 2 Protective effects of dietary supplementation with L Figure 2 Protective effects of dietary supplementation with L. crispatus M247 during DSS colitis. To induce colitis mice received 5% DSS in their drinking water for 5 days. After 24 h of DSS administration animals were randomly divided to receive daily (for 3 days) either live L. crispatus M247 (10⁴, 10⁶ or 10⁸ bacteria), 10⁸B. clausii spores or 10⁸ heat killed L. crispatus M247. Severity of colitis was monitored by measure of the body weight and results are shown as percent change over initial body weight (panel a). At the end of the experiment animals were killed, and full-thickness samples of the proximal colon were collected to determine MPO activity, expressed as U/mg wet tissue (panel b) or fixed in 10% PFA, paraffin embedded and sections stained with H & E were analyzed by a pathologist in a blinded fashion. Original magnification was 40× (panel c). Panel c shows a representative colonic section from a control mouse (left), a mouse receiving DSS alone (middle) or treated with 10⁸L. crispatus M247 (right). Mucosal ulceration and inflammatory cell infiltrate are drastically reduced in L. crispatus M247 — treated mice (compare II and III). Results are expressed as means ± SEM. ^*P < 0.05 vs control; ^**P < 0.01 vs control, ⁺P < 0.05 vs DSS alone; ⁺⁺P < 0.01 vs DSS alone. From: Beneficial effect of auto-aggregating Lactobacillus crispatus on experimentally induced colitis in mice FEMS Immunol Med Microbiol. 2005;43(2):197-204. doi:10.1016/j.femsim.2004.08.011 FEMS Immunol Med Microbiol | © 2004 Federation of European Microbiological Societies.

Figure 1 L. crispatus M247 adhering to colon mucosal surface by scanning electron microscopy. Full thickness colonic segments from normal mice were incubated 10 min with 10⁸ CFU/ml L. crispatus M247, then thoroughly washed with PBS and fixed with 2.5% glutaraldheyde. Sections were examined by a scanning electron microscope XL30 ESEM (Philips). Panel a shows a representative image of M247 cell adhering to colon mucosal surface and to mucus fibers (7000×), panel b shows clusters of M247 cells adhering to the mucosal surface (5000×). From: Beneficial effect of auto-aggregating Lactobacillus crispatus on experimentally induced colitis in mice FEMS Immunol Med Microbiol. 2005;43(2):197-204. doi:10.1016/j.femsim.2004.08.011 FEMS Immunol Med Microbiol | © 2004 Federation of European Microbiological Societies.