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Formation of Urolithins in Normal Volunteers Following Pomegranate Extract Consumption for 28 Days is Associated with Differences in the Gut Microflora Population

Although pomegranate ellagitannins are highly bioactive in vitro, they are not absorbed intact but are hydrolyzed in the intestine to ellagic acid and urolithins. There are known interindividual differences in the intestinal formation and urinary excretion of urolithin A (UA). To investigate the difference in gut microbiota between UA producers and non‐producers 20 healthy volunteers consumed pomegranate extract for 4 weeks. Urine and stool samples were collected at baseline and 4 weeks for urolithin analysis by HPLC and DNA sequencing with Illumina MiSeq analysis. The following differences (percent of total bacteria) in baseline gut microflora phyla were seen between UA producers (n=14) and non‐producers (n=6): Verrucomicrobia (genus Akkermansia) (0.2% vs.6.6%); Actinobacteria (5.2 %vs.1.8%); Firmicutes (68% vs.57%); Bacteroidetes (23%vs.26%) and the Firm/Bact ratio (2.96 vs. 2.19). The abundance of Proteobacteria was significantly increased (3.1% vs. 5.5%) and Actinobacteria decreased (1.8% vs 1.4%) comparing baseline to 4 weeks in producers. The production of urolithins was positively associated with the abundance of Akkermansia, a bacteria previously shown to be inversely correlated with body weight in rodents and humans. Probiotic bifidobacteria were generally not affected by pomegranate extract. Pomegranate extract consumption over 28 days induced changes in the microbiome to enhance production of urolithins but the profile of microflora was stable and different between producers and non‐producers. Future clinical studies of pomegranate juice and extract may benefit from stratifying between producers and non‐producers.