Synergistic Effect of a Novel Combination of Pre-, Pro-, and Postbiotics on Microbial Fermentation and Metabolite Production In Vitro

Abstract

Abstract Objectives To test the effect of probiotic strains (Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12; PS), a prebiotic (bacteriophage blend; BB), a postbiotic (tributyrin; TB), or a combination of all three (full mix; FM) on microbial fermentation and metabolite production in vitro. Methods Short term colonic incubation was performed under conditions representative of the proximal large intestine, using the fecal microbiota of a single healthy human donor. At the start of the experiment, treatments and a blank control (BC), were added to the reactors. Each reactor also received a carbohydrate-supplemented nutritional medium and freshly prepared fecal inoculum. Reactors were closed with a rubber septum, made anaerobic through flushing with nitrogen, and incubated for 48 hours. Each condition was run in triplicate. Overall fermentative activity (pH and gas production), changes in microbial metabolite production (short chain fatty acid, lactate, and ammonium analysis) were assessed. Results In the first 6 hours, microbial fermentation occurred in all treatments, indicated by pH shifts and production of gas and lactate. FM, PS, and BB stimulated production of lactate compared to BC (p < 0.05). After 24 hours, FM, PS and TB stimulated production of butyrate compared to BC (p < 0.05). FM and PS yielded more lactate than either BB or TB (p < 0.05). After 48 hours, FM, PS, BB, and TB each stimulated production of butyrate compared to BC (p < 0.05). Additionally, FM yielded significantly more butyrate than PS, TB, or BB (p < 0.05). Conclusions All treatments resulted in similar pH profiles and were representative of ideal conditions for bacterial growth and metabolism. The increased production of lactate in FM and PS was attributed to the fermentative activities of probiotics LGG and BB-12. Each treatment stimulated production of butyrate compared to BC but the largest increase in butyrate production was seen in FM, indicating a synergistic relationship between pre-, pro-, and postbiotics. Funding Sources The study was conducted by an independent lab, ProDigest, and funded and sponsored by Ritual (Natals, Inc.). Compound Solutions, Inc. also financially supported the study.