Microbial distribution at family level.

<p>Family level microbial distribution in all data sets at intervention week 2 and week 8. The pie charts represent total percentage read number for the corresponding colour coded family (<i>n</i> = 9–10 per group).</p

Delta weight gain over the eight week intervention period.

<p>(A) Bac⁺ intervention, when compared to Bac^- intervention, causes a significant reduction in weight gain in diet induced obese mice at weeks 2–4 (early intervention period) but this does not persist with time. (B) Vancomycin treatment results in a two phase reduction in weight gain in diet induced obese mice. In phase one (early; weeks 1–4) a significant reduction in weight gain relative to the initial start weight is observed. In the second phase, diet induced obese mice receiving vancomycin gain weight relative to the initial start weight but weight change continues to be significantly less than that in diet induced obese controls. Data represented as mean SEM n = 9–10 *p<0.05.</p

Microbial distribution at phylum level.

<p>Phylum level microbial distribution in all data sets at intervention week 2 and week 8. The pie charts represent total percentage read number for the corresponding colour coded phylum (<i>n</i> = 9–10 per group).</p

Total bacterial number observed in all treatment groups at both time points.

<p>Quantitative PCR reveals that the changes occurring are qualitative not quantitative as no significant difference is observed between time points. Total bacterial numbers calculated as copies of 16 S rRNA/g wet stool. Statistical significant difference between treatment groups is denoted by ***. p value based on kruskal wallis analysis with statistical significant determined as p≤0.05. Error bars represent the standard error of the mean.</p

Extraction of <i>Lactobacillus</i> flagellin proteins and analysis on 10% SDS-PAGE gels.

<p>A: Pilot extraction of surface proteins from motile and non-motile <i>L. ruminis</i> strains. The arrow points to the ATCC27782 flagellin protein. The larger protein at ∼34 kDa is GAPDH. B: Flagellin proteins extracted from various <i>Lactobacillus</i> species as indicated.</p

Flagellate <i>L. ruminis</i> cells or <i>Lactobacillus</i> flagellin stimulate IL8 production by intestinal epithelial cells in a TLR5-dependent manner.

<p>A: Flagellate <i>L. ruminis</i> ATCC27782 cells induce significant IL8 secretion from human intestinal epithelial cell lines. T84, n = 9; HT-29, n = 7; Caco-2, n = 6. The increments on the Y-axis follow a square-root scale. B: Following transfection with siRNA directed against TLR5, IL8 secretion from HT-29 cells was significantly lower in response to ATCC27782 whole bacteria. Boxplots are based on six experimental replicates. Box-plots show the median and the interquartile range. Solid black dots represent outlier values. Statistical significance was determined using a one-tailed Mann-Whitney U test. Horizontal black bars indicate significant differences between samples. The level of statistical significance is given by P-values over these bars, or by the following symbols: P<0.001 = ***; P<0.05 = *; P<0.1 = ▪.</p

Transmission electron micrographs of <i>L. ruminis</i> whole cells and flagella.

<p>A: <i>L. ruminis</i> ATCC27782; 1% ammonium molybdate, 6000× magnification. B: <i>L. ruminis</i> ATCC27782 flagellum filament with visible attached hook structure; 2% ammonium molybdate, 250000× magnification. C: <i>L. ruminis</i> ATCC25644; 0.25% ammonium molybdate, 15000× magnification.</p

Aggregate microbiota composition at phylum level in faecal samples from infants (panel a), mothers (panel b), at order/family level in infants (panel c) and mothers (panel d), and at <i>Bifidobacterium</i> genus level in the same sets of individuals (panels e and f), as indicated.

<p>In panels a and b only major taxonomic groups are shown.</p

Rarefaction curves generated for 16 S rRNA gene sequences obtained from stool samples of infants and faecal samples of mothers.

<p>Panel a represents the rarefaction curves using the Chao index. Panel b displays rarefaction curves using the Shannon index.</p

Number of sequences per phylotype for each sample.

<p>The y axis is a neighbour-joining phylogenetic tree containing one representative of each of the OTUs detected in this study; each row represents a different OTU. Increasing darkness of the grayscale corresponds to higher estimated relative abundance. The most conserved OTUs between samples are marked with an arrow. Arrow 1 represents the OTUs corresponding to <i>B. longum</i>, arrow 2 indicates the OTUs of <i>B. pseudocatenulatum</i>, arrow 3 highlights the phylotypes belonging to <i>Streptococcus thermophilus</i>. The boxed area includes those phylotypes belonging to <i>Ruminococcaceae</i> and <i>Clostridiales</i> taxa. The feeding method is indicated for each subject; Br, breast-fed; Bo, bottle fed.</p