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This study evaluated the effects of simulated gastrointestinal conditions (SGIC) on combined potentially probiotic Limosilactobacillus fermentum 296 (~ 10 log CFU/mL), quercetin (QUE, 160 mg), and/or resveratrol (RES, 150 mg) as the bioactive components of novel nutraceuticals. Four different nutraceuticals were evaluated during exposure to SGIC and analyzed the plate counts and physiological status of L. fermentum 296, contents and bioaccessibility of QUE and RES, and antioxidant capacity. Nutraceuticals with QUE and RES had the highest plate counts (4.94 ± 0.32 log CFU/mL) and sizes of live cell subpopulations (28.40 ± 0.28%) of L. fermentum 296 after SGIC exposure. An index of injured cells (Gmean index, arbitrary unit defined as above 0.5) indicated that part of L. fermentum 296 cells could be entered the viable but nonculturable state when the nutraceuticals were exposed to gastric and intestinal conditions while maintaining vitality. The nutraceuticals maintained high contents (QUE ~ 29.17 ± 0.62 and RES ~ 23.05 mg/100 g) and bioaccessibility (QUE ~ 41.0 ± 0.09% and RES ~ 67.4 ± 0.17%) of QUE and RES, as well as high antioxidant capacity (ABTS assay ~ 88.18 ± 1.16% and DPPH assay 75.54 ± 0.65%) during SGIC exposure, which could be linked to the protective effects on L. fermentum 296 cells. The developed nutraceuticals could cross along the gastrointestinal tract with high concentrations of functioning potentially probiotic cells and bioavailable phenolic compounds to exert their beneficial impacts on consumer health, being an innovative strategy for the co-ingestion of these bioactive components.

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This study evaluated the stability of a novel nutraceutical formulation composed of the probiotic Limosilactobacillus fermentum 296, quercetin (QUE), and resveratrol (RES) (LFQR) under different storage conditions. The effects of different relative humidities (RH; 11, 22, and 33%) and storage temperatures (refrigeration temperature -4 °C and room temperature -25 °C) on the stability of LFQR were evaluated through the determination of thermal stability, viable cell counts, bacterial physiological status, antioxidant capacity, and contents of QUE and RES during long-term storage. RH did not affect endothermic reactions and mass reduction in LFQR. After a 15-day-humidification period, L. fermentum 296 had higher viable cell counts in LFQR under refrigeration temperature storage when compared to room temperature storage regardless of the RH. The physiological status of L. fermentum 296 in LFQR was overall similar during 90 days of storage (11% RH) under refrigeration and room temperature. L. fermentum 296 had the highest viable cell counts (> 6 log CFU/g) in LFQR up to day 90 of refrigeration storage (11% RH). LFQR kept high contents of QUE and RES and maintained antioxidant capacity during 90 days of storage under refrigeration and room temperature. The results showed that the higher stability and functionality of LFQR during long-term storage should be guaranteed under 11% RH and refrigeration temperature.

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This study evaluated the impacts of different nutraceutical formulations combining Limosilactobacillus fermentum 296 (∼10 log CFU/mL), quercetin (QUE, 160 mg), and or resveratrol (RES, 150 mg) on the relative abundance of various intestinal bacterial populations, production of microbial metabolites, and antioxidant capacity during 48 h of in vitro colonic fermentation. The nutraceutical formulations increased the relative abundance of Lactobacillus spp./Enterococcus spp. and Bifidobacterium spp. and decreased the relative abundance of Bacteroides spp./Prevotella spp., Clostridium histolyticum, and E. rectale/C. coccoides during the colonic fermentation. Medium with the formulation containing L. fermentum, QUE, and RES had the highest prebiotic indexes, indicating synergistic or additive interaction between QUE and RES to modulate the intestinal microbiota. The nutraceutical formulations increased the production of bioactive metabolites and antioxidant capacity in the colonic fermentation media. The results indicate the capability of the tested nutraceutical formulations to beneficially modulate the composition and metabolite production of human intestinal microbiota and increase the antioxidant capacity in the intestinal environment.

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The increasing interest in the effects of the gut microbiota on host health has stimulated the investigation of the composition of this microbial community and the factors affecting these microorganisms. This review discusses the recent advances and progress applications in the use of the fluorescent in situ hybridization (FISH) coupled to flow cytometry (FC) technique (FISH-FC) in studies evaluating the gut microbiota published in the last 10 years, with particular emphasis on the effects of foods and dietary interventions. These studies have shown that FISH-FC technique is capable of detecting and quantifying several groups of bacteria found as part of the gut microbiota. FISH-FC can be considered an effective, versatile, and rapid technique to evaluate alterations in gut microbiota composition caused by different foods as assessed in studies in vitro, in vivo, and in clinical trials. Some specific probes have been most used to represent the general gut microbiota, such as those specific to Lactobacillus spp./Enterococcus spp., Bacteroidaceae/Prevotellaceae, Clostridium histolyticum, and Bifidobacterium spp. FISH-FC technique could have an important opportunity for application in studies with next-generation probiotics belonging to the gut microbiota. Optimizations of FISH-FC protocols could allow more discoveries about the gut microbiota, including the development of new probes targeting microorganisms still not explored, the analysis of individual portions of the intestine, and the proposition of novel quantitative approaches.

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