Figure 1.

Experimental Set-Up. Microbiotas were harvested from the feces of five participants (3 men and 2 women) as proxies of their gut microbiota. Samples were derived from homogenized human feces stabilized in 20% (w/v) anaerobic, pre-reduced 1× PBS/10% (v/v) glycerol/0.1% (w/v) l-cysteine. Samples were inoculated to 2% (w/v) in 1 mL of Mi-Pro optimized culture media and incubated under anaerobic conditions at 37 °C for 18 h. To evaluate the potential effects of BAFASAL^® on human gut microbiota, we compared (1) BAFASAL^®-treated microbiota (BAF) to those microbiotas exposed to (2) heat-inactivated BAFASAL^® (IBAF), (3) fructo-oligosaccharides as a positive control (FOS), and (4) phosphate-buffered saline (PBS; pH 7.6) as a negative control in triplicate and at two concentrations, as defined in materials and methods. Following culturing, microbial cells were pelleted and frozen until further analysis. DNA was extracted for metagenomic analyses using 16S rRNA gene-based sequencing and proteins extracted and digested for metaproteomic analysis, as described in materials and methods. Bioinformatic analysis was used to determine gut microbiome compositional and functional changes in response to treatment.