Biology and Life Sciences

Abstract: Background: Polyphenols interact bidirectionally with the gut microbiota and may enhance short‑chain fatty acid (SCFA) production, yet evidence from human randomized controlled trials (RCTs) has not been comprehensively synthesized. Objectives: This systematic review and meta‑analysis evaluated the effects of polyphenol supplementation on gut microbiota composition, microbial diversity, and fecal SCFA concentrations in adults, and examined moderators of these effects. Methods: Five databases were searched through October 2023 for RCTs assessing oral polyphenol supplementation in adults. Eligible studies reported outcomes related to gut microbiota composition or fecal SCFAs. Random‑effects meta‑analyses were conducted for SCFA outcomes, and subgroup analyses examined effects by polyphenol class, dose, duration, health status, and analytical methods. Risk of bias was assessed using the Cochrane RoB 2 tool, and certainty of evidence using GRADE. Results: Fifty RCTs (n = 2,042 participants) were included. Polyphenol supplementation increased total SCFAs in 70.6% of studies and significantly increased butyrate concentrations (pooled SMD = 0.48; 95% CI: 0.32–0.64; I² = 58%). Acetate and propionate increased in 75% and 71.4% of studies, respectively. A consistent shift toward a more butyrogenic fermentation profile was observed. Polyphenols enriched beneficial genera, including Bifidobacterium (81.8%), Akkermansia muciniphila (50%), and Faecalibacterium prausnitzii (45.5%), while reducing potentially pathogenic taxa such as Enterobacteriaceae and Clostridium spp. Improvements in alpha diversity were reported in 66.7% of studies, and beta diversity separation in 87.5%. Effects were stronger in individuals with metabolic disorders and in interventions lasting ≥12 weeks. Conclusions: Polyphenol supplementation consistently enhances beneficial gut bacteria, increases SCFA production, particularly butyrate, and improves microbial diversity in adults. These findings support classifying polyphenols as bioactive prebiotics that meet ISAPP criteria. This term underscores their distinction from traditional prebiotics within our proposed five-phase model: whereas fibers are characterized by their fermentability, polyphenols integrate direct antimicrobial activity against pathogenic species (e.g., Staphylococcus aureus, Enterobacteriaceae) with their role as selective substrates for beneficial microbes. This dual mechanism of action suggests that polyphenols do not merely supplement the microbiota but actively reshape it by pruning harmful taxa while fertilizing beneficial ones. Polyphenol-rich strategies represent promising microbiota-focused approaches; however, while shifts in microbial profiles coincide with better metabolic health, further research is required to bridge the correlation-causation gap and confirm whether these microbial changes directly drive clinical outcomes.

Abstract: The study examined whether cow's milk and beef differ from plant-based alternatives based on emotional profiling. It also sought to determine whether two panels of trained observers with different levels of preparation and experience would reach consistent results. Cow’s milk was compared with almond drink and oat drink, and beef with tofu and seitan. All products were organic. Two panels of trained observers (n=8/10, n=9/11) evaluated the products. The EmpathicFoodTest® (EFT) was used for emotional profiling. Cow’s milk, almond drink, and oat drink could be clearly distinguished from one another. Significant effects were observed in all 12 items of the EFT. Beef also differed significantly from the plant-based alternatives tofu and seitan, specifically in 8 and 9 individual characteristics, respectively. However, the differences between tofu and seitan were not significant. Milk and meat differed primarily in the items “warm/cold” and those related to emotional well-being compared to plant-based alternatives. The two independent panels, one with many years of experience, the other with brief, intensive preparation, achieved largely consistent results. Very good consistency was found for the warm/cold item, the duration item, and the items of emotional well-being. Emotional profiling can describe meat, milk, and plant-based alternatives. Intensive training over a short period enables a panel to achieve results comparable to those of a panel with several years of experience.

Abstract: Using a viability-resolved metagenomics protocol coupled with copy-number-corrected absolute quantification, we report three findings from a longitudinal study of eight ready-to-eat (RTE) seafood matrices subjected to combined pasteurization and high-pressure processing (HPP). First, we demonstrate that 16S amplicon relative abundances can be converted into absolute cell counts concordant with ISO culture methods (Lin's CCC = 0.94; Bland-Altman bias = −0.25 log₁₀, non-significant; 93% within ±1 log₁₀). This establishes that the three principal barriers to quantitative amplicon metagenomics — compositional bias, copy-number distortion, and dead-cell DNA contamination — can be jointly overcome within a single analytical workflow. Second, we provide the first community-scale quantification of viable but non-culturable (VBNC) populations in processed foods: 60% of indicator groups with negative culture harbored molecular loads of 10³–10⁷ cells/g. The discrepancy scaled with predicted membrane resilience — maximal for sporulating clostridia (+5.9 log₁₀), intermediate for Gram-negative Enterobacteriaceae (+4.0 log₁₀), minimal for lactic acid bacteria — implicating differential DNase impermeability as a biological mechanism rather than an algorithmic artifact. Third, we uncovered a post-treatment microbial succession qualitatively distinct from that described for HPP alone. Whereas HPP-only products are typically dominated by mesophilic Leuconostoc and Lactobacillus, the combined thermal-barometric treatment selected spore-formers (Clostridium spp., Sporosarcina), the thermo-barotolerant lactic acid bacterium Carnobacterium as dominant LAB, and post-process recontamination psychrotrophs (Psychrobacter, Acinetobacter). Combining lethal barriers thus reconfigures surviving community structure rather than merely reducing it. These findings were enabled by DELOREAN (Deep Estimation of Live Organisms by Read Equivalence to Agar Numbers), a framework that integrates an original sample preparation protocol — Viable Target Metagenomics (vtMG) — with a quantitative conversion algorithm. vtMG enriches DNA from membrane-intact cells through differential lysis and DNase treatment as an enzymatic alternative to PMA; the algorithm then anchors copy-number-corrected relative abundances to fluorimetric total DNA quantification via empirical scaling factors. Calibration factors were derived in a prior, independent pilot study and applied without modification to the validation dataset, constituting a legitimate out-of-distribution test. The framework is classifier-agnostic, platform-independent, and designed for progressive refinement through collaborative calibration.

Abstract: To investigated the effect of exogenous selenium on selenium enrichment and antioxidant activity of germinated chestnut. We treated ‘Zaofeng’ chestnuts with 0, 20, 40, 60 and 80 mg/L concentration of Na₂SeO₃, and the analyses focused on total Se, SeCys₂, MeSecys, Se^IV, SeMet, Se^VI, γ -aminobutyric acid (GABA), antioxidant enzyme (PAL, SOD, GPX, CAT) activity, non-enzymatic antioxidant substances (total polyphenols and flavonoids) content and antioxidant capacity (DPPH, ABTS) during germination. The results indicated that low concentration of selenium (20-40 mg/L) significantly promoted the organic transformation of selenium, increased antioxidant enzyme activity and phenol accumulation, and enhanced antioxidant capacity of chestnut. High concentration of selenium (80 mg/L) induced oxidative stress, inhibited enzyme activity and reduces antioxidant capacity of chestnut. During the germination of chestnut, selenite was absorbed by embryo and subsequently transformed into organic Se in vivo, ultimately being stored in the form of SeCys₂. The selenium enrichment rate decreased significantly with the increase of Na₂SeO₃ treatment concentration. Furthermore, the treatment with 40 mg/L Na₂SeO₃ led to a significant increase in GABA content of germinated chestnut. Overall, 20-40 mg/L Na₂SeO₃ were identified as the suitable concentration for germinated chestnut. This study provides a theoretical basis for developing functional food of selenium-enriched germinated chestnut.

Abstract: Fermentation quality is shaped not just by what microbes produce but by how they communicate. Cocoa pulp juice fermentation is governed by quorum sensing (QS) — bacterial autoinducers (AHLs, AI-2, peptides) and fungal mediators (farnesol, tyrosol) — that coordinate microbial succession and flavour precursor formation. Yet QS states are absent from operational fermentation control: conventional bioreactors act only on macroscopic variables (pH, temperature, dissolved oxygen) and cannot read the molecular language coordinating quality-determining transitions. This translational perspective proposes that a digital twin, implemented as a bioprocess decision suite, provides the missing intelligence layer. It is not a vessel replacement but a QS-aware reasoning system that translates communication states into explainable, audit-ready decisions and enables in silico sensory predesign before a run begins. A key design constraint is polyphenol-mediated AHL quenching and acid-accelerated AI-2 degradation in the cocoa matrix, which we formalise as an architectural specification. Time-series transformer models with attention-based explainability anchor the near-term AI layer, with graph neural networks, Bayesian uncertainty quantification, and reinforcement learning forming a progressive maturation roadmap. A three-tier framework scales deployment from artisanal to biomanufacturing operations.

Abstract: Natural α-amylase inhibitors from plant sources remain underexplored, whereas raisin polyphenols exhibit considerable inhibitory potential. In this study, raisin polyphenols were purified with D101 macroporous resin, and the adsorption process followed the pseudo-first-order kinetic model and the Langmuir isotherm model, indicating monolayer, exothermic adsorption. UHPLC-MS/MS analysis of the purified raisin polyphenols (PRP) identified 10 phenolic compounds, with ferulic acid, quercetin, and isoquercetin as the major constituents. The inhibitory mechanism of PRP against porcine pancreatic α-amylase (PPA) was systematically investigated using inhibition kinetics, fluorescence spectroscopy, circular dichroism (CD) spectroscopy, molecular docking, and molecular dynamics (MD) simulations. PRP exhibited strong inhibitory activity (IC50=3.15±0.16 mg/mL) and acted as a mixed-type inhibitor, whereas the major monomers showed distinct inhibition behaviors. Fluorescence and CD analyses indicated that PRP and its monomers altered the microenvironment around tryptophan and tyrosine residues, induced secondary-structure rearrangement in PPA, and reduced enzymatic activity. MD simulations confirmed the stability of the polyphenol-PPA complexes. Binding free-energy calculations showed that quercetin had the most favorable overall binding affinity and complex stability among the tested monomers, driven mainly by hydrogen bonding and hydrophobic interactions. These findings clarify the α-amylase inhibitory mechanisms of PRP and its key constituents and provide a theoretical basis for developing natural hypoglycemic functional ingredients and plant-derived α-amylase inhibitors.

Abstract: 4-Ethylguaiacol (4-EG) is an important volatile phenolic compound that contributes characteristic smoky, woody, and spicy notes to fermented foods and beverages, particularly Baijiu. In this study, a 4-EG-producing strain, designated JN11, was obtained by screening isolates from Baijiu pit mud and identified as Bacillus coagulans based on morphological, physiological, biochemical, and 16S rRNA analyses. In sorghum juice medium, strain JN11 produced 271.57 ± 2.73 μg/L 4-EG. To elucidate its role in volatile phenol formation, the phenolic acid decarboxylase gene, BcPAD, was cloned and heterologously expressed in Escherichia coli BL21(DE3). The BcPAD gene comprises 504 bp and encodes a 167-amino-acid protein. Recombinant BcPAD exhibited maximal activity at pH 6.0 and 50 °C and retained more than 60% residual activity after 5 h at 30–40 °C. Fe³⁺ increased enzyme activity to 115.36% of the control, whereas Zn²⁺ markedly inhibited enzyme activity and SDS completely inactivated the enzyme. BcPAD showed the highest activity toward p-coumaric acid, with a specific activity of 460.55 ± 18.30 U/mg and a catalytic efficiency (K_cat/K_m) of 6.26 ± 0.75 mM^-1 • s^-1, while lower activities were observed toward caffeic acid and ferulic acid, and no activity was detected toward sinapic acid. Homology modeling and molecular docking further indicated that the superior catalytic performance toward p-coumaric acid was associated with favorable hydrogen-bonding interactions and a productive binding orientation within the active site. These findings provide new insights into phenolic acid decarboxylases from B. coagulans and support the potential applications of strain JN11 and BcPAD in volatile phenol biosynthesis and flavor modulation during Baijiu production.

Abstract: Spent mushroom substrate (SMS), the main by-product of mushroom production, is rich in valuable compounds that could be recovered by ultrasound-assisted extraction (UAE) and exploited as fat-mimetic functional ingredients in food formulations. In this study, low-fat cookies prototypes were developed by incorporating a dietary fiber extract obtained from SMS using UAE. The extraction process was optimized following a Box–Behnken experimental design, identifying optimal conditions at a specific energy input of 200 J/mL, a particle size of 2 mm, and a solute-to-solvent ratio of 1:27, yielding a dietary fiber recovery of 30.82%. The optimized SMS extract exhibited high oil-holding capacity (1.39 g/g), emulsion stability (80%), and foaming capacity (83.55%). Four cookie formulations were evaluated, among which G1 (50% fat replacement) showed the best balance between consumer acceptability and an improved nutritional profile, characterized by higher protein (8.4 g/100 g), total dietary fiber (7.10 g/100 g), and mineral contents. Notably, G1 cookies displayed a significant reduction in predicted glycemic index, decreasing from 83.84 in the control to 69.65. Overall, these results demonstrate that optimized SMS-derived dietary fiber is an effective functional ingredient for the development of low-fat, high-fiber, and reduced-glycemic cookies, contributing to the valorization of agro-industrial by-products within a circular economy framework.

Abstract: Given the increasing relevance of sustainability certification in food supply chains and, at the same time, rising confusion among consumers about the multitude of labels on food products, concerns about the value of sustainability certification occur frequently. This paper aims to investigate consumers’ evaluation and purchase intentions, and willingness-to-pay (WtP) for blockchain-enabled sustainability certification in coffee. Utilizing a questionnaire guided by an extended model of Ajzen’s theory of planned behavior (TPB), an online survey was conducted with n = 400 German consumers. Data were analyzed using structural equation modeling and cluster analysis. The results revealed perceived behavioral control (PBC) and SN as the most influential factors on WtP, whereas intention to buy is shaped by PBC and environmental concerns. Three distinct clusters were identified with concise preference, intention, and WtP profiles, highlighting heterogeneous consumer motivations. The study is the first one exploring the topic in the German context. It provides novel insights for academics and industry stakeholders on sustainable coffee, blockchain technology, and factors influencing evaluation and purchase intentions for coffee with blockchain-enabled sustainability certifications. It also paves the way for future research on blockchain-enabled sustainability certification in other product groups. Moreover, the paper contributes to a more nuanced theoretical understanding of the TPB and its applications.

Abstract: Fresh-cut vegetables are perishable products, which means they are prone to rapid physiological and microbiological degradation, leading to a quick deterioration in appearance, a decline in the organoleptic and nutritional properties, and increased losses. Therefore, new solutions are being sought to ensure safety and extend their shelf life. This review focuses on the quality and shelf-life stability of fresh-cut vegetables that have undergone heat shock treatment before or after minimal processing. The aim was to collect and present studies showing the beneficial effects of thermal shock on fresh-cut vegetables. Most studies have been conducted using hot water as a heat carrier, but hot air and steam have also been tested. Experimental data showing the combined effects of thermal shock and other physical or chemical methods are also presented. The collected data serve as guidelines for future research aiming to optimize methods and for use in the minimal processing of vegetables on an industrial scale.

Abstract: Ancient leavened bread is said to have been developed in Egypt. We hypothesized that microorganisms inherent in wheat flour were involved in fermentation of bread. The dough, made only with wheat flour and water, was kept warm for a day before being baked. The dough began to ferment after 12 hours, and when baked after 24 hours, it yielded bread with a specific volume of 1.7-2.0 cm3/g. Microorganisms were isolated from the dough before baking and identified using a rapid microbial identification mass spectrometry system. In spelt flour, Kosakonia cowanii was the dominant species. Numerous Pantoea agglomerans were isolated from strong flour B, followed by the detection of Moraxella osloensis. A considerable amount of the Gram-positive bacterium Bacillus cereus was detected from medium flour. These bacteria can be harmful to the human body. However, the high temperatures involved in the bread-baking process can potentially reduce the number of live bacteria.

Abstract: The growing consumption of plant-based meat alternatives (PBMAs) has increased attention to their microbiological safety, particularly under refrigerated storage conditions. Although the PBMA market has expanded rapidly, data on the microbiological status of industrially produced, heat-treated products remain limited. The present study aimed to evaluate, within a descriptive framework, the microbiological safety of industrially produced, heat-treated PBMAs during refrigerated storage. A total of 100 PBMA formulations, including salami-type and frankfurter-type ready-to-eat products, were manufactured under standardized industrial conditions and subjected to validated thermal processing (core temperature ≥ 92 °C for varying durations). Microbiological analyses were conducted at four predefined storage intervals (day 0, day 15, day 35, and day 60 at 0˗4 °C) to assess the presence of selected foodborne pathogens (Salmonella spp. and Listeria monocytogenes) and hygiene indicator microorganisms (generic Escherichia coli, Enterobacteriaceae, coagulase-positive Staphylococcus aureus, and Bacillus cereus). Intrinsic physicochemical parameters relevant to microbial survival and growth, including pH, water activity, moisture content were also determined. Salmonella spp. and Listeria monocytogenes were not detected (absence in 10 g) in any sample at any storage time point. Hygiene indicator microorganisms were not detected during early storage (day 0-15), while limited occurrence was observed at extended storage (day 60), including Escherichia coli (3%), coagulase-positive Staphylococcus aureus (20%), and Bacillus cereus (15%). Detected Staphylococcus aureus levels ranged between 103 and 105 CFU/g. These findings indicate strong microbiological stability during early refrigerated storage, with limited microbial occurrence at extended storage intervals (day 60). Overall, the evaluated products demonstrated a favorable microbiological safety profile under the applied processing and storage conditions. Given formulation heterogeneity and the absence of biological replication, findings are interpreted descriptively and provide an industrially relevant safety overview rather than inferential conclusions.

Abstract: This study reports on the design and characterisation of thermo-reversible gelatine hydrogels incorporating beeswax-structured oil-in-water emulsions as novel 3D-printable food inks. Beeswax oil-in-gel emulsions (BOGEs) were prepared by varying the sunflower oil to beeswax (SFO:BW) mass fraction (1:0, 3:1, 1:1, 1:3 and 0:1) at a fixed lipid loading (10% wt.) within a 4% wt. gelatine matrix. The BOGEs were evaluated in terms of microstructure, thermophysical properties, small and large amplitude oscillatory shear rheological behaviour, instrumental hardness, and 3D printability. CLSM images revealed a progressive transition from an emulsion-filled to a bigel-like microstructure with increasing beeswax content, driven by partial crystallisation and percolation of lipid droplets. Differential scanning calorimetry confirmed that beeswax incorporation progressively suppressed the gelatine hydrogel fusion enthalpy, indicating that wax crystal lattices govern the supramolecular organisation of the gelatine network. SAOS tests showed that BW enhanced the elastic modulus, with a critical solid fat content threshold (Φc = 0.294) above which lipid droplet percolation provided an additional structural reinforcement. LAOS characterisation revealed a type III nonlinear viscoelastic response, with delayed yielding and enhanced structural integrity at higher BW fractions. Instrumental hardness measurements confirmed the active filler role of BW at mass fractions ≥0.5. 3D printing assessment demonstrated that intermediate SFO:BW ratios (3:1 and 1:1) afforded the highest printing fidelity, combining favourable extrusion flow with adequate post-deposition shape retention. Overall, the results demonstrate that beeswax-structured emulsions can effectively tailor the structure–function properties of gelatine hydrogels, enabling the development of clean-label, multiphase food inks suitable for extrusion-based 3D printing applications.

Abstract: Food fermentation is a widely used processing technique that enhances sensory properties, shelf life, and nutritional value, partly through the activity of beneficial microorganisms. This study investigated the microbial communities associated with traditional pearl millet fermentation and their potential nutritional contributions. Pearl millet (TiftLHB open-pollinated variety) was obtained from USDA-ARS and subjected to spontaneous fermentation in sterilized water at 28 ± 2 °C for 72 hours, followed by wet milling and an additional 72-hour fermentation. Microbial DNA was extracted, and 16S rRNA amplicon sequencing was performed after PCR amplification and quality control. Sequence data were analyzed using DADA2 and PICRUSt2 pipelines for taxonomic and functional prediction. The dominant bacterial genera identified were Weissella and Lactobacillus, both commonly associated with cereal fermentations. Weissella is known for reducing antinutrients and contributing to folate production, while the overall microbial profile was consistent with reports from other regions, including the presence of lactic acid bacteria such as Leuconostoc. These findings suggest that spontaneous fermentation of pearl millet supports microbial communities with potential nutritional and functional benefits. Metagenomic approaches may provide an effective strategy for identifying and optimizing beneficial microorganisms to enhance the nutritional quality and health-promoting properties of fermented cereal-based foods.

Abstract: This study evaluates animal welfare as a factor influencing economic losses and re-source use efficiency in the global livestock sector for 2025-2026. Using analytical as-sessment and conceptual modeling (TEL, WFW, and LDI models) based on FAO and USDA data, the study estimates the economic and environmental implications of pre-slaughter stress. The results suggest that global meat losses may reach approximately 19.4 million tons annually, with associated economic losses potentially exceeding USD 90 billion. These losses correspond to an estimated 184.2 km³ of freshwater and approximately 138.5 million tons of feed resources, indicating reduced efficiency in resource utilization. The findings highlight that improving animal welfare may represent not only an ethi-cal consideration but also a potential approach to enhancing resource efficiency and sustainability in livestock production systems. The study supports the integration of welfare-related parameters into agricultural and food system policies, particularly in regions affected by logistical disruptions.

Abstract: S. Tomé and Principe (STP) islands have been studied in recent years for their wide range of medicinal plants which exhibit several biological activities of great medicinal interest for some diseases. Experimental planning for optimization of several parameters was carried out by a full factorial of two levels of three factors for secondary metabolite extraction from Tithonia diversifolia leaves by using water and hexane at 25 and 40 ºC and 150 rpm for 0 and 5 days of incubation. The best conditions for highest extraction of phenolic compounds (i.e 72.16 mmoles gallic acid equivalent/g leaves)) was obtained at 40ºC, in H₂0 and 5 days of incubation. Several phytochemical assays were performed for characterization of these plant extracts and the highest levels for extraction of reducing power, ABTS and DPPH were obtained at 25 ºC, H₂0 and 5 days of incubation whereas highest levels of SOD activity were extracted at 40ºC, H₂0 and 5 days of incubation. The present report consists of a novel and intrinsic synchronous fluorescence and phosphorescence characterization of secondary metabolites from this plant extract. Intrinsic and non-destructive synchronous fluorescence was carried out in the range of 250 to 750 nm with a Δλ range of 5–30 nm which exhibited peaks at 290, 320, 345, 400, 490 and 675 nm in hexane plant extracts whereas aqueous extracts revealed only peaks at 490, 560 and 675 nm. On the other hand, intrinsic and non-destructive synchronous phosphorescence was also performed which exhibited peaks at 325, 400, 490, 550, 675 nm and 500, 560 nm, respectively. 3-D spectra of secondary metabolites confirmed the peaks at 290, 320, 345, 400, 490 and 675 nm in plant extracts. FTIR spectroscopy was selected to investigate the structural properties of secondary metabolites in these plant extracts. Therefore, the present work describes a novel characterization of secondary metabolites by a non-destructive and intrinsic synchronous fluorescence techniques for plant extracts.

Abstract: Cachaça is a sugarcane spirit produced exclusively in Brazil and is the second most consumed alcoholic beverage by the Brazilian population. During the maturation process, carried out either in wooden barrels or with addition of wood chips, compounds such as organic acids and phenolic substances are extracted into the beverage, providing specific characteristics to the matured spirit. The present study objectivated to evaluate the physicochemical properties, phenolic composition, sensory profile, consumer acceptability and ranking preference of blended cachaças produced with cachaça aging with jackfruit wood chips (Artocarpus heterophyllus), with or without aeration, with white cachaça. The results showed that samples blended with a cachaça stored in stainless steel tanks containing wooden chips (SSW) presented higher values of total acidity, volatile acidity, total esters, dry extract, and total phenolic compounds when compared with cachaça stored under conventional conditions (CWB). Experts described the cachaça blends as having a balanced/harmonious flavor, moderate woody, alcohol and acidity, and a mild spicy. No significant differences were observed in consumer acceptability between treatments. However, the blended cachaças prepared with samples stored in stainless steel tanks containing wooden chips, showed greater preference among consumers. Therefore, blending unaged cachaça with cachaça aged with wooden chips appears to be a promising and advantageous alternative for the cachaça production process, since the physicochemical and sensory characteristics are preserved.

Abstract: Metabolic dysfunction-associated steatotic liver disease (MASLD) affects 25%–30% of adults globally. Dietary sugar reduction is one of the key therapeutic targets, but elimination of sugar-sweetened foods may compromise adherence to calorie-restricted diets. Brazzein, a natural sweet protein that is 500–2000 times sweeter than sucrose, offers a promising substitute, yet clinical data in patients with MASLD are lacking. In a double-blind, randomized, two-period crossover trial, 103 adults with MASLD tasted iso-sweet vanilla ice cream sweetened with either brazzein or sucrose on two consecutive days. Overall impression and sensory attributes (appearance, color, aroma, flavor, and texture) were rated on 5-point hedonic scales, and the percentage of the 100 g portion consumed was recorded. Brazzein-sweetened ice cream met the prespecified criteria for both non-inferiority and equivalence versus sucrose for overall impression. Top-2 box acceptance (ratings ≥4) was extremely high and nearly identical (96.1% for brazzein and 98.1% for sucrose). Mean consumption exceeded 98% of the portion for both products, with no significant difference between sweeteners. Secondary sensory ratings were closely similar, and multivariate analyses indicated highly overlapping sensory profiles. Exploratory subgroup analyses suggested consistent findings across most demographic and clinical characteristics, although participants with advanced liver fibrosis (LSM ≥9.6 kPa) showed numerically higher ratings for sucrose. Liver stiffness was the only significant predictor of consumption, with slightly lower intake at higher stiffness values. This study provides the first evidence that brazzein-sweetened ice cream maintains consumer acceptability comparable to a conventional sucrose-sweetened product in a clinically relevant population, supporting its potential integration into dietary management strategies for MASLD.

Abstract: This study aimed to evaluate how specific quantities of green tea and sugar, as well as fermentation temperature, impact variations in kombucha quality parameters. The study used tea quantities ranging from 0.5 to 3.0% w/v, sugar from 3.0 to 6.0% w/v, and fermentation temperatures of 20°C and 26°C. Beverage quality parameters such as pH, volatile acidity, alcohol content, and SCOBY growth were evaluated. At a temperature of 20 ± 2°C, formulations with 6.0% w/v sugar showed no SCOBY growth, and two for-mulations showed volatile acidity above the established maximum limit of 130 mEq L⁻¹. Most formulations had an alcohol content below 0.5% v/v and were classified as non-alcoholic. At 26 ± 2°C, the greatest SCOBY growth occurred, with the highest rec-orded value of 203%. Only two formulations showed an alcohol content above 0.5% v/v, but with values close to the limit. High amounts of sugar do not favor SCOBY growth at either mild or higher temperatures (26°C). Variations in temperature and ingredient quantities influence the production of green tea kombucha that meets safety and quality requirements. These data show the variations in ingredients and temperatures that favor kombucha production, considering its quality and classification.

Abstract: The growing demand for functional foods has stimulated the development of chocolate matrices enriched with bioactive ingredients. This study aimed to optimize the extraction of phenolic compounds using hydro-organic solvents of different polarities, characterize the phenolic and alkaloid profile of distinct chocolate formulations by HPLC-DAD, and evaluate whether fortification with microencapsulated vitamins E and D, zinc, and omega-3 fatty acids (EPA and DHA), either alone or combined with partial replacement of cocoa butter by a structuring oil, enhances the extractable bioactive compounds and in vitro antioxidant activity. Five formulations were evaluated: control chocolate (C), choc-olate containing vitamin microcapsules (T1), chocolate with DHA/EPA microcapsules (T2), lipid-modified chocolate with structuring oil (T3), and chocolate combining microcapsules with lipid modification (T4). Phenolic compounds were extracted using 50% ethanol, 70% methanol, and 70% acetone. Among the tested solvents, 70% methanol showed the highest extraction efficiency, enabling broader detection of bioactive compounds. HPLC-DAD analysis revealed compounds characteristic of cocoa-based matrices, including epicatechin, gallic acid, vanillin, and traces of procyanidins, as well as the methylxanthine alkaloids theobromine and caffeine. The T4 formulation showed a greater abundance of extractable compounds compared to the other formulations. Antioxidant activity was evaluated using DPPH radical scavenging and β-carotene/linoleic acid bleaching assays. T4 exhibited the highest antioxidant performance, with 21.8% DPPH inhibition (EC₅₀ = 2.5 mg/mL) and the highest optical density in the β-carotene assay, indicating enhanced protection against lipid peroxidation. These findings demonstrate that the combination of microencapsula-tion and lipid phase modification improves antioxidant functionality and supports the development of value-added functional chocolates.