Figure 3
Eugenol treatment when combined with metronidazole prevents mucus thickening and highlights the importance of microbial factors.
(a), AB/PAS stained methanol-Carnoy's fixed distal colon sections showing the inner mucin layer (white arrowheads). i = inner mucus layer. Original magnification = 400×. (b), Quantification of inner mucus layer thickness. Distal colon sections stained with AB/PAS to visualize and quantify the inner mucus layer. n = 5–10 mice per group. (*p = 0.038, ***p = 0.0003, ***p = 0.0001) U = untreated mice. Significance determined using two-tailed Student's t-test and expressed as the mean ± SEM. (c), Simpson's Reciprocal Index of diversity was used to determine diversity of fecal communities after treatment with eugenol, metronidazole, metronidazole and eugenol or untreated mice. n = 4–8. U = untreated mice. (**p = 0.0048). (d), Similarity between the microbiota of untreated and eugenol treated feces was measured by Bray-Curtis index and depicted in a dendogram. U = untreated, E = eugenol treated. (e), Family level phylogenetic classification of 16S rRNA gene frequencies in feces collected from untreated, eugenol, metronidazole or combined treatment. Those indicated with a classification level other than family level could only be identified confidently to the level indicated. Classification scheme: k, kingdom; p, phylum; c, class; o, order; f, family. Representative data is shown for each group, n = 4–8. Stars indicate significant family changes between eugenol and untreated mice (p < 0.05).
