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Background/Objectives : Diet significantly influences gut microbiota (GM), with variations in GM responses linked to the type and quantity of food consumed. These variations underscore the need for personalized nutrition. The Stance4Health (S4H) project developed the S4H Food Frequency Questionnaire (S4H-FFQ) and the i-Diet S4H app to assess dietary intake of foods affecting GM. This study aimed to validate the S4H-FFQ against the validated I.Family-FFQ and the i-Diet S4H app; Methods: The S4H-FFQ, with 200 food items across 14 food groups, evaluates dietary intake over the past month. Qualitative validation compared food group consumption frequencies from the S4H-FFQ and the I.Family-FFQ, while quantitative validation assessed nutrient and energy intake using the i-Diet S4H app. The S4H-GM score, a measure of GM-relevant food consumption, was evaluated through the S4H-FFQ and i-Diet S4H app; Results : Pearson correlations between the S4H-FFQ and the I.Family-FFQ ranged from 0.3 to 0.7 and were statistically significant across all the food groups. Quantitative validation showed lower but consistent correlations, comparable with other studies, confirming the S4H-FFQ's ability to estimate food intake. A positive correlation was also found between the S4H-GM scores from the S4H-FFQ and the i-Diet S4H app ( p < 0.001); Conclusions : The S4H-FFQ is a reliable tool for assessing dietary patterns that influence GM. Its application in nutritional studies can enhance personalized nutrition and support future research aimed at optimizing GM and improving health outcomes.

The gastrointestinal digestion of food and further gut microbial activity render a myriad of different molecules that could be responsible for the biological activities that are classically assigned to their parent compounds. This has been previously shown for some phytochemicals whose antioxidant capacity was either increased or decreased after being metabolized by gut microbes. Whether a global antioxidant capacity that is extracted from food is determined by the gut microbial community structure is still not well described. In the present study, we in vitro digested and fermented 48 different foods that were submitted to different culinary treatments using the stools of lean children, obese children, celiac children and children with an allergy to cow's milk proteins. Their antioxidant capacities were assessed with the DPPH and FRAP assays, and the percentage that each food contributed to their daily antioxidant intake as well as their antioxidant capacity by portion size was inferred. Overall, cereals, fruits and vegetables displayed a higher contribution to their daily antioxidant intake, while tubers, fish and meat exhibited a higher antioxidant capacity by serving size. The food that was fermented in the lean children's and those children that were allergic to cow's milk protein's fecal material, showed a higher antioxidant capacity, which could imply that there is a larger role of the gut microbiota in this area.

The prevalence of obesity has been increasing in children over the last few decades, becoming a concern for health professionals and governments. Gut microbial community structure in obese people have been found to differ from that of lean subjects for some taxa which could result in different production of microbial metabolites. The aim of the present work was to study whether the gut microbiota from obese children extracts a different concentration of antioxidant capacity than the gut microbiota from lean children. For this purpose, different foods were in vitro digested and in vitro fermented using fecal material from obese and lean children. FRAP, DPPH and Folin-Ciocalteu methods were used to measure the antioxidant capacity released during digestion and fermentation. Overall, when using lean gut microbiota, antioxidant capacity released was higher when measured via DPPH and FRAP. Moreover, according to DPPH results, lean gut microbiota could potentially release more antioxidant power from vegetables than from animal products, while obese gut microbiota did the opposite. On the contrary, with the FRAP method obese gut microbiota released higher levels of antioxidant power from plant products than from animal products, but the final antioxidant capacity was still lower than that released by lean gut microbiota. Therefore, these results reflect that the total antioxidant capacity of foods is influenced by the gut microbiota, although whether that antioxidant capacity is released from plant or animal products can be slightly influenced by the method used for analysis.

The gut microbiota has a profound effect on human health and is modulated by food and bioactive compounds. To study such interaction, in vitro batch fermentations are performed with fecal material, and some experimental designs may require that such fermentations be performed with previously frozen stools. Although it is known that freezing fecal material does not alter the composition of the microbial community in 16S rRNA gene amplicon and metagenomic sequencing studies, it is not known whether the microbial community in frozen samples could still be used for in vitro fermentations. To explore this, we undertook a pilot study in which in vitro fermentations were performed with fecal material from celiac, cow's milk allergic, obese, or lean children that was frozen (or not) with 20% glycerol. Before fermentation, the fecal material was incubated in a nutritious medium for 6 days, with the aim of giving the microbial community time to recover from the effects of freezing. An aliquot was taken daily from the stabilization vessel and used for the in vitro batch fermentation of lentils. The microbial community structure was significantly different between fresh and frozen samples, but the variation introduced by freezing a sample was always smaller than the variation among individuals, both before and after fermentation. Moreover, the potential functionality (as determined in silico by a genome-scaled metabolic reconstruction) did not differ significantly, possibly due to functional redundancy. The most affected genus was Bacteroides , a fiber degrader. In conclusion, if frozen fecal material is to be used for in vitro fermentation purposes, our preliminary analyses indicate that the functionality of microbial communities can be preserved after stabilization.