Figure 1.

Methodology to isolate extracellular vesicles, EVs, and characterization of the bacterial community. Diagram showing the methodology used to isolate EVs in vitro, based on sequential centrifugation and density gradient separation (A). Growth curve of fecal inoculum cultivated in vitro in the presence of β-mannan or without any carbohydrate (control) (B). pH measurement of in vitro cultures either with or without the presence of β-mannan (C). Microbial diversity and composition in the cultures containing either 2% β-mannan or no carbohydrate addition. Samples were collected at 0, 6, and 24 h and subjected to amplicon sequencing of the 16S rRNA gene. The relative abundances of the dominant phyla (D), orders (E), and genera (F) that resulted from in vitro cultivation in the absence (0% mannan) or presence (2% mannan) of β-mannan are shown as stacked bar plots. The black bars in D–F indicate the summed abundance of different phyla, orders, or genera that were individually present at less than 1% abundance.