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Coffee husks are the main by-product of the coffee industry and have been traditionally discarded in the environment or used as fertilizers. However, recent studies have shown that coffee husks have bioactive compounds, such as phenolics and fiber-bound macro antioxidants, offering a range of potential health benefits. This study evaluated the antioxidant capacity, cytoprotective/cytotoxic properties, and stimulatory effects on the relative abundance of selected intestinal bacterial populations of individuals with diabetes of organic coffee husks. Organic coffee husk had good antioxidant capacity, maintained under simulated gastric conditions, with more than 50% of antioxidant capacity remaining. Organic coffee husk exerted cytoprotective properties in Caco-2 cells, indicating that cellular functions were not disturbed, besides not inducing oxidation. Overall, organic coffee husk promoted positive effects on the abundance of distinct intestinal bacterial groups of individuals with diabetes during in vitro colonic fermentation, with a higher relative abundance of Bifidobacterium spp., indicating the availability of components able to reach the colon to be fermented by intestinal microbiota. Organic coffee husk could be a circular material to develop new safe and pesticide-free functional ingredients with antioxidant and potential beneficial effects on human intestinal microbiota.

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This study developed and evaluated chitosan-sodium alginate capsules containing the probiotic Lacticaseibacillus rhamnosus GG using extrusion and emulsification techniques. The encapsulated L. rhamnosus GG cells were also evaluated for technological and probiotic-related physiological functionalities, as well as when incorporated in UHT and powdered milk. Extrusion (86.01 ± 1.26%) and emulsification (74.43 ± 1.41%) encapsulation techniques showed high encapsulation efficiency and high survival rates of L. rhamnosus GG during 28 days of refrigeration and room temperature storage, especially emulsification capsules (> 81%). The encapsulated L. rhamnosus GG cells showed high survival rates during exposure to simulated gastrointestinal conditions (72.65 ± 1.09-114.15 ± 0.44%). L. rhamnosus GG encapsulated by extrusion and emulsification performed satisfactorily in probiotic-related physiological (pH and bile salts tolerance) and technological properties (positive proteolytic activity, diacetyl and exopolysaccharides production, high NaCl tolerance (> 91%), besides having high heat tolerance (> 76%)). L. rhamnosus GG in extrusion and emulsification capsules had high survival rates (> 89%) and did not significantly affect physicochemical parameters in Ultra-High Temperature (UHT) and powdered milk during storage. The results demonstrate that L. rhamnosus GG can be successfully encapsulated with alginate-chitosan as a protective material through extrusion and emulsification techniques. UHT and powdered milk could serve as appropriate delivery systems to increase the intake of this encapsulated probiotic by consumers.

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This study evaluated the use of acerola (Malpighia glabra L., CACE), cashew (Anacardium occidentale L., CCAS), and guava (Psidium guayaba L., CGUA) fruit processing coproducts as substrates to promote the growth, metabolite production, and maintenance of the viability/metabolic activity of the probiotics Lactobacillus acidophilus LA-05 and Lacticaseibacillus paracasei L-10 during cultivation, freeze-drying, storage, and exposure to simulated gastrointestinal digestion. Probiotic lactobacilli presented high viable counts (≥8.8 log colony-forming units (CFU)/mL) and a short lag phase during 24 h of cultivation in CACE, CCAS, and CGUA. Cultivation of probiotic lactobacilli in fruit coproducts promoted sugar consumption, medium acidification, and production of organic acids over time, besides increasing the of several phenolic compounds and antioxidant activity. Probiotic lactobacilli cultivated in fruit coproducts had increased survival percentages after freeze-drying and during 120 days of refrigerated storage. Moreover, probiotic lactobacilli cultivated and freeze-dried in fruit coproducts had larger subpopulations of live and metabolically active cells when exposed to simulated gastrointestinal digestion. The results showed that fruit coproducts not only improved the growth and helped to maintain the viability and metabolic activity of probiotic strains but also enriched the final fermented products with bioactive compounds, being an innovative circular strategy for producing high-quality probiotic cultures.

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Multi-strain Limosilactobacillus (L.) fermentum is a potential probiotic with reported immunomodulatory properties. This study aimed to evaluate the composition, richness, and diversity of the gut microbiota in male and female rats after treatment with a multi-strain of L. fermentum at different doses. Thirty rats (fifteen male and fifteen female) were allocated into a control group (CTL), a group receiving L. fermentum at a dose of 108 CFU (Lf-108), and a group receiving L. fermentum at a dose of 1010 CFU (Lf-1010) for 13 weeks. Gut microbiota and serum cytokine levels were evaluated after L. fermentum treatment. Male CTL rats had a lower relative abundance of Bifidobacteriaceae and Prevotella and a lower alpha diversity than their female CTL counterparts (p < 0.05). In addition, male CTL rats had a higher Firmicutes/Bacteroidetes (F/B) ratio than female CTL rats (p < 0.05). In female rats, the administration of L. fermentum at 108 CFU decreased the relative abundance of Bifidobacteriaceae and Anaerobiospirillum and increased Lactobacillus (p < 0.05). In male rats, the administration of L. fermentum at 1010 CFU decreased the F/B ratio and increased Lachnospiraceae and the diversity of the gut microbiota (p < 0.05). The relative abundance of Lachnospiraceae and the alpha-diversity of gut microbiota were negatively correlated with serum levels of IL1ß (r = -0.44) and TNFα (r = -0.39), respectively. This study identified important changes in gut microbiota between male and female rats and showed that a lower dose of L. fermentum may have more beneficial effects on gut microbiota in females, while a higher dose may result in more beneficial effects on gut microbiota in male rats.

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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 investigated the potential impacts of the flour from Cereus jamacaru cactus cladodes (CJF), a cactus native to the Brazilian Caatinga biome, on the growth and metabolism of different potentially probiotic strains, as well as on the abundance of selected intestinal bacterial populations and microbial metabolic activity during in vitro colonic fermentation with a pooled human fecal inoculum. Cultivation of the probiotics in a medium with C. jamacaru cladodes flour (20 g/L) resulted in viable cell counts of up to 9.8 log CFU/mL, positive prebiotic activity scores (0.73-0.91), decreased pH and sugar contents, and increased lactic, acetic, and propionic acid production over time, indicating enhanced probiotic growth and metabolic activity. CJF overall increased the relative abundance of Lactobacillus spp./Enterococcus spp. (2.12-3.29%) and Bifidobacterium spp. (4.08-4.32%) and decreased the relative abundance of Bacteroides spp./Prevotella spp. (8.35-6.81%), Clostridium histolyticum (6.91-3.59%), and Eubacterium rectale/Clostridium coccoides (7.70-3.95%) during 48 h of an in vitro colonic fermentation using a pooled human fecal inoculum. CJF stimulated the microbial metabolic activity, with decreased pH, sugar consumption, lactic and short-chain fatty acid production, alterations in overall metabolic profiling and phenolic compound contents, and maintenance of high antioxidant capacity during colonic fermentation. These results show that CJF stimulated the growth and metabolic activity of distinct potential probiotics, increased the relative abundance of beneficial intestinal bacterial groups, and stimulated microbial metabolism during in vitro colonic fermentation. Further studies using advanced molecular technologies and in vivo experimental models could forward the investigation of the potential prebiotic properties of CJF.

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This study formulated sweet potato chips with powdered potentially probiotic Levilactobacillus brevis (SPLB) and Lactiplantibacillus plantarum (SPLP) and evaluated their impacts on human intestinal microbiota during 48 h of in vitro colonic fermentation. L. brevis and L. plantarum kept high viable cell counts (> 6 log CFU/g) on sweet potato chips after freeze-drying and during 60 days of storage. SPLB and SPLP had satisfactory quality parameters during 60 days of storage. SPLB and SPLP increased the relative abundance of Lactobacillus ssp./Enterococcus spp. (3.84-10.22%) and Bifidobacterium spp. (3.25-12.45%) and decreased the relative abundance of Bacteroides spp./Prevotella spp. (8.56-2.16%), Clostridium histolyticum (8.23-2.33%), and Eubacterium rectale/Clostridium coccoides (8.07-1.33%) during 48 h of in vitro colonic fermentation. SPLB and SPLP achieved high positive prebiotic indexes (> 8.24), decreased pH values and sugar contents, and increased lactic acid and short-chain fatty acid production, proving selective stimulatory effects on beneficial bacterial groups forming the intestinal microbiota. The results showed that SPLB and SPLP have good stability and high viable cell counts of L. brevis and L. plantarum when stored under room temperature and caused positive impacts on human intestinal microbiota, making them potentially probiotic non-dairy snack options.

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This study evaluated the impacts of novel nutraceuticals formulated with freeze-dried jabuticaba peel (FJP) and three potentially probiotic Limosilactobacillus fermentum strains on the abundance of bacterial groups forming the human intestinal microbiota, metabolite production, and antioxidant capacity during in vitro colonic fermentation. The nutraceuticals had high viable counts of L. fermentum after freeze-drying (≥ 9.57 ± 0.09 log CFU/g). The nutraceuticals increased the abundance of Lactobacillus ssp./Enterococcus spp. (2.46-3.94%), Bifidobacterium spp. (2.28-3.02%), and Ruminococcus albus/R. flavefaciens (0.63-4.03%), while decreasing the abundance of Bacteroides spp./Prevotella spp. (3.91-2.02%), Clostridium histolyticum (1.69-0.40%), and Eubacterium rectale/C. coccoides (3.32-1.08%), which were linked to positive prebiotic indices (> 1.75). The nutraceuticals reduced the pH and increased the sugar consumption, short-chain fatty acid production, phenolic acid content, and antioxidant capacity, besides altering the metabolic profile during colonic fermentation. The combination of FJP and probiotic L. fermentum is a promising strategy to produce nutraceuticals targeting intestinal microbiota.

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This study investigated the effects of freeze-dried red beet root (FDBR) and freeze-dried red beet stem and leaves (FDBSL) on target bacterial groups and metabolic activity of human colonic microbiota in vitro. The capability of FDBR and FDBSL to cause alterations in the relative abundance of different selected bacterial groups found as part of human intestinal microbiota, as well as in pH values, sugar, short-chain fatty acid, phenolic compounds, and antioxidant capacity were evaluated during 48 h of in vitro colonic fermentation. FDBR and FDBSL were submitted to simulated gastrointestinal digestion and freeze-dried prior to use in colonic fermentation. FDBR and FDBSL overall increased the relative abundance of Lactobacillus spp./Enterococcus spp. (3.64-7.60%) and Bifidobacterium spp. (2.76-5.78%) and decreased the relative abundance of Bacteroides spp./Prevotella spp. (9.56-4.18%), Clostridium histolyticum (1.62-1.15%), and Eubacterium rectale/Clostridium coccoides (2.33-1.49%) during 48 h of colonic fermentation. FDBR and FDBSL had high positive prebiotic indexes (>3.61) during colonic fermentation, indicating selective stimulatory effects on beneficial intestinal bacterial groups. FDBR and FDBSL increased the metabolic activity of human colonic microbiota, evidenced by decreased pH, sugar consumption, short-chain fatty acid production, alterations in phenolic compound contents, and maintenance of high antioxidant capacity during colonic fermentation. The results indicate that FDBR and FDBSL could induce beneficial alterations in the composition and metabolic activity of human intestinal microbiota, as well as that conventional and unconventional red beet edible parts are candidates to use as novel and sustainable prebiotic ingredients.

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Limosilactobacillus (L) fermentum (strains 139, 263, 296) is a novel probiotic mixture isolated from fruit processing by-products. The use of this formulation has been associated with improvements in cardiometabolic, inflammatory, and oxidative stress parameters. The present study evaluated the safety of a potential multi-strain probiotic by genotoxicity (micronucleus assay) and subchronic toxicity study (13-week repeated dose). In the genotoxicity evaluation, L. fermentum 139, 263, 296 did not increase the frequency of micronuclei in erythrocytes of rats of both sexes at doses up to 1010 CFU/mL. In the subchronic toxicity study, the administration of L. fermentum did not promote adverse health effects, such as behavioral changes, appearance of tumors, changes in hematological and biochemical parameters. In addition, higher doses of L. fermentum 139, 263, 296 have been shown to reduce the levels of pro-inflammatory cytokines. Administration of potentially probiotic L. fermentum did not promote adverse health effects in rats and could be evaluated as a potential probiotic for humans.