Food & Function

Traditional Mongolian fermented foods have been extensively utilized for dietary regulation and the promotion of gastrointestinal health. However, spontaneous fermentation remains inherently unpredictable, leading to significant variations in microbial community dynamics, metabolite accumulation, and the consistency and quality of the final product. Drawing on the traditional preparation of Mongolian acidic foods, this study established a controlled production strategy for whole-wheat probiotic fermented soup (WWPFS) by combining enzymatic pretreatment with probiotic-directed fermentation. Physicochemical characterization, 16S rRNA gene-based microbial community profiling, LC-MS/MS-based untargeted metabolomics, safety evaluation, and an Escherichia coli-induced gut microbiota dysbiosis model were employed to optimize and comprehensively characterize the fermentation process of WWPFS. The optimized process established a reproducible fermentation system consistently dominated by Lactobacillus and Bacillus across independent fermentation batches. Compared with traditional spontaneous fermentation, probiotic-directed fermentation remodeled the physicochemical properties of the whole-wheat matrix, including carbon, nitrogen, phosphorus, sulfur, and mineral composition, and facilitated the accumulation of putatively annotated LC-MS/MS features, including DL-lactate, 1,4-D-xylobiose, diacetyl, and phenyllactic-acid-related features derivatives. Acute oral and 28-day repeated-dose toxicity evaluations showed no treatment-related adverse effects within the tested dose range and study duration. In the Escherichia coli-induced gut microbiota dysbiosis mouse model, microbial richness, diversity, and community structure differed among the experimental groups, and both low- and high-dose WWPFS groups showed significant shifts in overall gut microbial community composition relative to the model group after multiple-testing correction, together with directional recovery of selected model-responsive bacterial genera. Cross-system integration identified coordinated response patterns between fermentation-derived metabolite features and model-responsive gut bacterial taxa, supporting a potential metabolite-microbiota link in WWPFS-mediated gut microbiota modulation. In summary, probiotic-directed fermentation improved the controllability of the traditional Mongolian fermented food production process, reshaped its metabolic profile, and enhanced its potential to modulate the gut microbiota. These findings provide experimental evidence supporting the modernization of traditional Mongolian fermented foods and the development of probiotic-based functional foods.

Nisin is a promising antimicrobial peptide widely used in food preservation due to its efficacy against Gram-positive spoilage and pathogenic bacteria. Although Nisin is increasingly applied in the food sector, the biopeptide suffers from instability within food matrixes and can rapidly lose its antimicrobial potential following interaction with food biomolecules. Thus, it is necessary to investigate approaches that can be employed to extend the stability and activity of Nisin. Hence, the aim of this study was to develop and characterise a chitosan-alginate polyelectrolyte microencapsulation system capable of enhancing Nisin stability while retaining antimicrobial activity. The microencapsulation of Nisin was achieved by pre-gelation of alginate using calcium chloride and subsequent direct electrostatic interaction between cationic Nisin and chitosan with pre-gelled anionic alginate at pH 5.0. Following microcapsule formation, physicochemical and structural characterisation was performed using Zeta potential determination and measurement of the polydispersity index (PDI) via dynamic light scattering. SEM micrographs were used to confirm morphology, while Fourier-transform infrared (FTIR) spectroscopy and high-performance liquid chromatography (HPLC) were utilised to assess chemical integrity and functional group preservation of encapsulated Nisin. Following this, stable microcapsules with diameters ranging from 150-200 nm and smooth surface morphology were obtained. Microcapsule formation was strongly influenced by formulation parameters, particularly pH, calcium ion concentration, and chitosan content, with deviations from optimal acidic conditions (< pH 5.0) resulting in aggregation, increased polydispersity, and reduced encapsulation efficiency. The microcapsules were monodispersed (PDI {approx} 0.30) and electrostatically stable, exhibiting a Zeta potential of approximately +36 mV. These microcapsules remained stable over a prolonged storage period of 21 days under refrigerated conditions while retaining antimicrobial activity against Bacillus cereus. Encapsulation efficiency reached approximately 65%, confirming effective retention of Nisin within the polymer matrix. Overall, the findings demonstrate that chitosan-alginate ionic gelation is a non-denaturing and effective encapsulation strategy for extending the functional stability of Nisin. These microcapsules show strong potential as natural antimicrobial delivery systems for food and beverage applications, particularly in acidic food matrices, with implications for improved food safety and shelf-life extension.

Biopreservatives including nisin and its derivatives are becoming more desirable in the food processing industry because of the growing demand for naturally preserved and minimally processed foods free from artificial preservatives. However, ensuring microbiological safety while meeting these consumer preferences remains a major challenge. This has necessitated the continuous investigation of potential new antimicrobial agents produced by naturally occurring microorganisms. Hence, this study explored the synthesis, characterisation, and optimisation of a bacteriocinogenic lactic acid bacterium and its antimicrobial product, possibly novel bacteriocin (Nisin 2A) from Lactococcus lactis isolated from commercial brined cheese. The isolation was achieved by screening for wild-type bacteriocin-producing lactic acid bacteria from dairy products using MRS media. Screening was performed using antagonism assays, yielding five producer organisms. Of these, the isolate whose metabolites exhibited the most potent antimicrobial activity was identified as Lactococcus lactis, which synthesised an active antimicrobial peptide designated as Nisin 2A, with a molecular mass of approximately 3.3 kDa as determined by UHPLC-MS and SDS-PAGE. Production of Nisin 2A was scaled up through fed-batch fermentation of Lactococcus lactis in modified MRS broth following process optimisation using a Plackett-Burman experimental design and purified by ammonium sulphate precipitation and solid-phase extraction (SPE). Furthermore, the antimicrobial potential of the bacteriocin was evaluated by the agar well diffusion assay and quantified using the tube dilution method. The purified peptide demonstrated broad-spectrum antimicrobial activity, particularly against the test Gram-positive bacteria Bacillus cereus and retained its bioactivity across a wide pH range (3-9) and high thermal conditions (up to 100 {degrees}C). Furthermore, it had high sensitivity to proteolytic enzymes (Proteinase K and Trypsin). Notably, the peptide was thermostable and retained up to 90% of its initial activity after thermal treatment and maintained consistent inhibitory performance after extended storage. These findings highlight the potential application of Nisin 2A as a natural biopreservative in food systems.

BackgroundEmerging evidence suggests that PFAS can cross blood-brain barrier and lead to neurotoxicity. Recent evidence also suggest that PFAS can bioaccumulate in gut microbiota resident in the gut. However, how gut microbes influence PFAS-driven reorganization of metabolic homeostasis in the brain is poorly understood. MethodsTo address this gap, we investigated how gut microbiota influences brain metabolomic and lipidomic responses to PFAS exposure. Specific pathogen-free (SPF) and germ-free (GF) mice were fed an obesogenic diet for 8 weeks to promote metabolic disturbance. After 1 week of acclimation, half received control water and half received water containing a PFAS mixture (PFHxS, GenX, PFOA, PFOS, and FTOH mixture). Plasma and brain samples (cortex, subcortex, cerebellum, olfactory bulb, and brainstem) were collected after 8 weeks. Untargeted analyses were performed for lipidomic, metabolomic and PFAS using high resolution liquid chromatography tandem mass spectrometry (LC-MS/MS). Data was processed using MassCube with open-sources libraries. ResultsPFHxS, GenX, PFOA, PFOS, PFDA, and PFDS were detected in plasma. PFHxS, PFOA, PFOS, and PFDS were detected across all five brain regions, with PFOS as the predominant brain-enriched species. Pathway analysis identified nicotinate and nicotinamide metabolism as the most consistently PFAS-altered pathway in both SPF and GF mice. PFAS exposure induced region-specific metabolic remodeling, with gut microbiota differentially modulating responses in the cortex, cerebellum, and brainstem, whereas the olfactory bulb showed a largely microbiota-independent response. In addition to local effects within individual brain regions, plasma-brain analysis suggested systemic metabolic responses across tissues, with association strength varying by brain region and microbiome status. Gut microbiota also shaped PFAS-induced lipid dysregulation in the brain, and methylnicotinamide and delta-valerobetaine were among the most responsive metabolites. ConclusionThis study is the first to demonstrate that resident microbiota impact PFAS-associated metabolic remodeling across the gut-plasma-brain axis. HighlightsO_LIPFAS-induced metabolic remodeling in the brain is modified by gut microbiota. C_LIO_LIPFAS exposure alters nicotinate and nicotinamide metabolism throughout the brain. C_LIO_LIPFAS-induced brain metabolic responses are region specific and microbiota dependent. C_LIO_LIPlasma-brain analysis suggests potential systemic metabolic disruption by PFAS. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=104 SRC="FIGDIR/small/743341v1_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@15301deorg.highwire.dtl.DTLVardef@9fac0aorg.highwire.dtl.DTLVardef@d7f0f4org.highwire.dtl.DTLVardef@10c29c2_HPS_FORMAT_FIGEXP M_FIG C_FIG

Kakadu plum (Terminalia ferdinandiana), an Australian native fruit, is among the richest known dietary sources of vitamin C and hydrolysable tannins, yet its capacity to protect the intestinal epithelium against oxidative stress remains largely unexplored. This study optimised the extraction of bioactive compounds from freeze-dried Kakadu plum powder and evaluated their antioxidant activity using both chemical and cellular antioxidant in vitro assay. Phenolic compounds were extracted using three solvents (water, 80% ethanol, and 80% methanol) combined with shaking, ultrasound, or microwave assistance. Solvent, rather than processing technique, was the dominant determinant of antioxidant capacity: ethanol and methanol maximised total phenolic content, total flavonoid content, and DPPH radical-scavenging activity, whereas water extracts showed the highest ferric-reducing antioxidant power. Twenty-four phenolic compounds identified by HPLC-ESI-QTOF-MS/MS were mapped by network pharmacology to nine core oxidative-stress targets, and cross-species molecular docking predicted conserved binding of key phenolics to canine orthologs of PTGS2 and MMP2. In an H2O2-induced oxidative-stress in vitro cell model using canine small intestinal epithelial cells, both water (less than 25 ug/mL) and ethanol (less than 250 ug/mL) extracts significantly suppressed intracellular reactive oxygen species (ROS) in a dose-dependent manner, with the ethanol extract effective across a wider concentration range. This work demonstrated that Kakadu plum extract could be a promising natural, multi-target antioxidant ingredient for canine intestinal health, and provided a reference for future in vivo research.

Kinema is a traditional fermented soybean food indigenous to the eastern Himalayan regions of Nepal and India. The fermentation process is primarily mediated by the bacterium Bacillus subtilis, which produces several bioactive compounds and enzymes with potential therapeutic applications. Considering the growing burden of cardiovascular diseases and the need for effective fibrinolytic agents for thrombolytic therapy, this study aimed to extract, partially purify, and evaluate the thrombolytic potential of kinemakinase derived from kinema prepared from white soybeans. Partial purification of the enzyme was achieved using ammonium sulfate precipitation. Thrombolytic activity was assessed in vitro using human blood clots, where three enzyme dilutions demonstrated clot lysis ranging from 66% to 68%, indicating considerable fibrinolytic potential. In silico analyses were also performed to investigate the structural and functional characteristics of the enzyme. The tertiary structure obtained from UniProt was modeled using the Robetta server and refined with GalaxyRefine. Docking with fibrin using ClusPro 2.0 and molecular dynamics simulations using iMODS confirmed favorable interaction and structural stability, while disulfide engineering enhanced protein stability. The findings suggest that kinema-derived kinemakinase may serve as a promising alternative thrombolytic agent, warranting further biochemical characterization and dosage optimization.

Plant-based diets may benefit planetary and human health. However, the molecular pathways linking plant-based diet quality to chronic diseases remain unclear. In 4,372 participants from the European Prospective Investigation into Cancer and Nutrition, we assessed 7,285 SomaScan-measured aptamers to identify circulating proteins associated with healthful (hPDI) and unhealthful (uPDI) plant-based dietary patterns using complementary statistical and machine-learning approaches. We then applied cis-pQTL Mendelian randomization (MR) and colocalization to prioritize diet-associated proteins with genetic evidence for associations with overall and site-specific cancers, type 2 diabetes (T2D), and cardiovascular disease (CVD). Among diet-associated proteins with MR evidence, 8 hPDI- and 12 uPDI-related protein-disease associations showed strong colocalization, including EGFR-breast cancer, MMP10-endometrial cancer, NCAN-T2D, and PCSK9-CVD. These findings identify candidate proteins that may link plant-based diet quality to chronic diseases and provide biological insights into the potential health benefits of healthful plant-based diets, which are increasingly relevant to public and planetary health.

Mycosporine-like amino acids (MAAs) are among nature's most effective sunscreen compounds, capable of converting harmful ultraviolet radiation into harmless heat, and are widely distributed in marine organisms such as red macroalgae. Although decades of research have led to numerous discoveries, the rate of new MAA identifications has declined. To address this, we considerably expanded our previously developed combinatorial MAA database, increasing the number of covered structures tenfold. Following a comprehensive literature search for plausible but undescribed building blocks, the database now incorporates an extensive set of proteinogenic and non-proteinogenic amino acids, as well as other marine organic osmolytes, in combination with all (currently) known MAA scaffolds. This expanded resource was integrated into our identification platform, which combines UHPLC-VWD-HRMS2 analysis, feature-based molecular networking, and bioinformatics-driven annotation. Application of this updated workflow enabled the isolation and structural elucidation of a novel MAA, mycosporine-cysteinolic acid, from the red marine macroalga Vertebrata lanosa. Altogether, this study provides a valuable extension of the bioinformatics-based MAA screening pipeline, enhancing the annotation and discovery of novel MAAs in natural matrices.

High dietary fructose consumption is a major contributor to the development of obesity and related cardiometabolic diseases, highlighting the need for accurate assessment of fructose metabolism in humans. Stable isotope tracer approaches, such as 13C6-fructose, require highly sensitive and specific analytical methods to quantify both concentrations and isotopic enrichments. In this study, we developed and validated a robust gas chromatography-triple quadrupole mass spectrometry (GC-QQQ)-based method for the simultaneous measurement of unlabeled and 13C6-fructose in human plasma. The method employs oximation and per-acetate derivatization, and demonstrates high specificity and accuracy. Intra- and inter-assay precision were below 10%, with no detectable carry-over, and a lower limit of quantification (LLOQ) of 0.1 nmol/mL for concentration and 0.02 molar percent excess (MPE%) for enrichment and no interference from glucose. We further compared data acquisition using multiple reaction monitoring (MRM) and selected ion monitoring (SIM). MRM showed superior performance at the low concentrations and enrichment levels characteristic of clinical plasma samples, resulting in improved sensitivity and lower LLOQs compared to SIM. Overall, this validated method provides a sensitive and reliable approach for fructose tracer studies in humans. Its application will facilitate robust investigations into fructose metabolism, and its role in metabolic dysregulation and obesity-related disease.

Green synthesis of zinc oxide nanoparticles (ZnO NPs) offers a sustainable strategy for developing multifunctional antimicrobial nanomaterials. In this study, ZnO NPs were synthesized using Azadirachta indica leaf extract and characterized by UV-vis spectroscopy, FTIR, XRD, SEM, and GC-MS. The nanoparticles exhibited a characteristic absorption peak at 352 nm, a direct band gap of 3.07 eV, and hexagonal wurtzite crystallinity with an average crystallite size of approximately 32 nm. The biosynthesized ZnO NPs showed concentration-dependent antibacterial activity against Erwinia carotovora, producing inhibition zones of up to 25.9 mm. Mechanistic studies revealed significant membrane damage, evidenced by 4.77-fold and 5.62-fold increases in extracellular protein and amino acid leakage, respectively, with marked alterations in bacterial protein profiles detected by SDS-PAGE. The nanoparticles also exhibited strong antioxidant activity, achieving 89.4% DPPH radical scavenging, and induced dose-dependent cytotoxicity in HepG2 cells with an estimated IC50 of 124.8 g/mL. These findings demonstrate that neem-mediated ZnO nanoparticles possess potent antibacterial activity through membrane disruption while exhibiting promising antioxidant properties, highlighting their potential as eco-friendly nanomaterials for the management of bacterial soft rot and other phytopathogenic diseases.

Importance: Food and non-alcoholic beverage marketing drives children's dietary intake, yet updated evidence quantifying effects by marketing medium and sociodemographic factors is needed to inform policy. Objective: To quantify the effect of food marketing on dietary intake among children and adolescents (0-19 years) and examine variations by age, sex, socioeconomic position (SEP), weight status, marketing medium, and exposure duration. Data Sources: Nineteen electronic databases were searched for articles published from April 2020 to February 2026, complemented by World Health Organization-commissioned reviews covering 1970 to March 2020. Study Selection: Two reviewers independently selected peer-reviewed primary studies that assessed the association between food marketing and dietary intake, following PRISMA guidelines, with no language restrictions. Data Extraction and Synthesis: Two reviewers independently extracted data and assessed the risk of bias. Random-effects meta-analyses were conducted. The certainty of evidence was assessed using GRADE. Main Outcomes and Measures: Dietary intake (energy, quantity, or number of items consumed). Results: A total of 55 studies (N = 6,877; range 2-18 years) were included. Food marketing was associated with higher dietary intake (mean difference [MD], 34.8 kcal; 95% CI, 20.2-49.4) compared with no or less marketing. Unhealthy marketing via television (20 studies; MD, 44.5 kcal; 95% CI, 11.2-77.8), digital media (11 studies; MD, 37.5 kcal; 95% CI, 20.1-54.9), and packaging (11 studies; MD, 20.5 kcal; 95% CI, 0.7-40.3) all increased intake; the difference across media was significant (p < .001). Higher intake was observed in males (3 studies; MD, 51.9 kcal; 95% CI, 45.4-58.3) but not in females (MD, -6.8 kcal; 95% CI, -60.3-46.6); difference was not significant (p = .082). Differences by weight status (p = .012) were seen (5 studies; normal weight: MD, 55.6 kcal; 95% CI, -51.3-162.5; overweight/obese: 146.9 kcal; 95% CI, 34.1-259.7). Effects varied by age (p = .003) and by digital media exposure duration (p = .044). One study examined ethnicity; none studied SEP. Conclusions and Relevance: Food marketing is associated with increased dietary intake, with low certainty of evidence. Variations were observed across age, sex, weight status, and marketing medium. Further research is needed for adolescents and the role of SEP.

Dairy-free foods must reproduce diverse functions of dairy, from foam stabilization in a latte to acid-gel structure in yogurt and melt-stretch behavior in mozzarella. Public consumer benchmarks rarely compare these categories under a common protocol. Here we analyze data from the NECTAR Taste of the Industry 2026 study, a blinded, randomized, in-person evaluation of commercial dairy-free products and dairy benchmarks. The full study included 2,183 consumers and 112 products across ten categories, with 3,220 product evaluations and 79,027 recorded survey responses, including 35,762 product-linked responses. The ten categories included barista milk, butter, cheddar, cream cheese, creamer, ice cream, fluid milk, mozzarella, sour cream, and Greek style yogurt. Within each category, we compare a dairy benchmark with the dairy-free product selected post hoc as the category leader. Participants rated overall liking, flavor, texture and mouthfeel, appearance, similarity to dairy, and purchase intent on seven-point scales and completed check-all-that-apply questions. In exploratory category-specific equivalence tests using a chosen margin of {+/-}0.05 points, barista milk, creamer, and fluid milk meet this margin. Mozzarella has the largest paired deficit of -1.46 points (unadjusted, p < 0.001), followed by Greek style yogurt and butter; all three remain significant after Holm correction across the ten primary comparisons. Descriptive penalty analysis associates off-flavor, artificial or chemical notes, stale or cardboard notes, bitter taste, and lingering aftertaste with the largest reductions in liking. Together, these results establish the first open sensory benchmark for dairy and dairy-free products, demonstrate that sensory parity is already achievable in several beverage categories, and identify the product categories and sensory attributes that offer the greatest opportunities for future innovation.

BackgroundAtherosclerosis is driven by macrophage foam cell formation resulting from excessive oxidized low-density lipoprotein (oxLDL) accumulation and chronic vascular inflammation. This study evaluated the therapeutic potential of Aegeline, Atorvastatin, and their combined in mitigating oxLDL-induced inflammatory responses, cholesterol accumulation, and oxLDL uptake in human THP-1 macrophages. MethodsTHP-1 monocytes were differentiated into macrophages using a 72-hour differentiation protocol followed by a 48-hour resting period, confirmed via CD14 surface marker characterization. Macrophages were exposed to DiI-oxLDL and treated with Aegeline, Atorvastatin, or their combination. Key inflammatory cytokines and chemokines (CRP, TNF-, IL-6, and IL-8) were measured using ELISA. Cholesterol efflux capacity and cellular oxLDL uptake were quantitatively assessed using fluorescence retention assays and immunofluorescence imaging. ResultsDifferentiation of THP-1 monocytes to macrophages resulted in marked down-regulation of CD14 expression. DiI-oxLDL exposure triggered significant pro-inflammatory mediator secretion (p<0.001) and excessive intracellular cholesterol accumulation. Single-agent treatment with Aegeline or Atorvastatin significantly attenuated oxLDL-induced elevations of CRP, TNF-, IL-6, and IL-8. Atorvastatin alone strongly suppressed CRP expression back to physiological baseline levels (p=ns vs. control). Notably, the combination of Aegeline and Atorvastatin demonstrated enhanced, broad-spectrum anti-inflammatory efficacy, achieving superior suppression of TNF- (p=ns vs. control), IL-6, and IL-8 compared to monotherapies. Furthermore, both agents promoted cholesterol efflux and suppressed oxLDL uptake, with the combination treatment producing the lowest residual intracellular cholesterol levels (p<0.001). ConclusionAegeline and Atorvastatin effectively suppress oxLDL-induced macrophage inflammatory cascades and intracellular lipid overload. While Atorvastatin monotherapy exerts robust control over CRP and oxLDL loading, combining Aegeline with Atorvastatin provides synergistic efficacy, enhancing cholesterol efflux and restoring pro-inflammatory cytokine expression toward physiological levels. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=132 SRC="FIGDIR/small/744794v1_ufig1.gif" ALT="Figure 1"> View larger version (50K): org.highwire.dtl.DTLVardef@1d90d88org.highwire.dtl.DTLVardef@1079202org.highwire.dtl.DTLVardef@2d659org.highwire.dtl.DTLVardef@4685af_HPS_FORMAT_FIGEXP M_FIG C_FIG

Copper is an essential redox-active micronutrient, but agricultural soils are increasingly contaminated by copper from mining, industrial discharge, and intensive agrochemical use, pushing concentrations beyond levels plants can tolerate. Excess copper triggers Fenton-like reactive oxygen species (ROS) generation, mitochondrial dysfunction, and impaired growth. Existing mitigation strategies, such as soil amendments, phytoremediation, antioxidants, and different chelators, have been explored to reduce copper toxicity, but their effectiveness can be limited by immobilization, poor specificity, and environmental persistence. The present work introduces a nanoparticle-based strategy for the direct sequestration of excess copper coupled with protection against the oxidative damage caused by copper stress. Here, we report MPA-iron oxide nanoparticles (MIONPs), sequentially functionalized with chitosan, glutathione, and 3-mercaptopropionic acid, designed to simultaneously scavenge ROS, restore redox homeostasis, and chelate copper via surface thiol groups. MIONPs showed a significant increase in copper binding capacity over bare iron oxide nanoparticles (BIONPs) and, in copper-stressed Solanum lycopersicum seedlings, significantly improved germination and root/shoot growth, reduced intracellular ROS, restored mitochondrial membrane potential, and preserved nuclear integrity. This integrated design establishes MIONPs as a promising, dual-function nanoplatform for sustainable copper stress management in agriculture.

Edible mushrooms have been reported to have antimicrobial properties and other health benefits. This study aims to test the antimicrobial activities of several edible mushrooms from markets and test if co-culturing them with bacteria could induce stronger anti-bacterial properties. Commercial mushrooms, Hericium erinaceus (lions mane), Pleurotus ostreatus (oyster mushroom), Lentinula edodes (Shiitake) and Agaricus bisporus (button mushroom), were grown from strictly controlled/sterile substrates. Ethanol and water extracts from the mushrooms were prepared and tested against the bacteria Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis, and the fungus Candida albicans for antimicrobial activities. Shiitake water extract (SWE) showed strong antibacterial effects against all tested bacterial species, inhibitory effects on their biofilms, and antifungal activity. The antimicrobials in SWE seem to damage the cell wall and cell membrane of the bacteria, prefer weak acidic conditions, and are heat labile. Some antimicrobials are likely proteins and polysaccharides. In contrast, 3 other mushrooms displayed only weak antimicrobial effects. The fast-growing lions mane and oyster mushroom were co-cultured with different bacteria. The co-cultivation promoted the fruiting body development of lions mane. Co-culturing with S. aureus increased the anti-bacterial effects of lions mane against S. aureus, E. coli and particularly B. subtilis. Co-culturing the oyster mushroom with bacteria, especially B. subtilis and P. aeruginosa, boosted the mushroom growth. All tested bacteria, especially S. aureus, increased oyster mushroom anti-bacterial effect against E. coli and B. subtilis. The findings indicate that mushroom-bacteria co-culturing could have benefits both agriculturally and medicinally.

Purpose: Iron deficiency impairs sports performance, and female athletes are particularly vulnerable. High-dose iron supplements, commonly used to prevent iron depletion and performance impairments, may cause gastrointestinal side effects and disrupt the gut microbiota. Whether lower doses can improve iron status without adverse effects remains unclear. The aim of this study was to characterize iron intake and iron status in female soccer players during the competitive season and investigate effects of low-dose iron supplementation on iron status, safety-related outcomes and gut microbiota. Methods: In a two-arm parallel randomized controlled trial, female soccer players (median age 21) were randomized to an intervention group (n=12) receiving 3-month low-dose iron supplementation (27 mg elemental iron/day) or a control group (n=11) without supplementation. Blood/fecal samples were collected at baseline and 3-month follow-up. Dietary intake was estimated using 7-day food diaries. Between-group differences were analyzed per protocol (n=18) using ANCOVA with baseline adjustment. Results: The players had inadequate baseline iron intake (median 11.2 mg/day) and 43% had serum ferritin indicating iron depletion (<35 g/L). At follow-up, no significant between-group difference was found for serum ferritin, but fewer athletes in the intervention group experienced decreases from baseline to follow-up (P<0.05). Moreover, serum iron was higher in the intervention group (Pgroup=0.05). No between-group differences were observed for gastrointestinal symptoms or liver damage biomarkers. On the contrary, the intervention led to lower IL-6 (Pgroup=0.04) and higher gut microbial -diversity (Pgroup=0.01) compared to controls. Conclusions: The low-dose iron supplementation was well tolerated, attenuated decreases in iron stores, increased gut microbial diversity and attenuated systemic inflammation in female soccer players with suboptimal dietary iron intake. However, potential adverse effects of long-term exposure cannot be excluded.

Romanian bors, a traditional fermented wheat bran beverage, is produced through spontaneous fermentation and represents a complex microbial ecosystem. Despite its cultural importance and presumed health benefits, its microbial ecology and functional potential remain poorly characterized. The present study aimed to elucidate the microbial community structure of bors and link it to functional traits relevant to fermentation performance and food functionality by integrating culture-independent sequencing with culture-dependent isolation and functional characterization. A total of 32 bors samples (12 commercial and 20 homemade) were analyzed. Amplicon-based sequencing revealed a microbiome dominated by lactic acid bacteria (LAB), with lactobacilli accounting for the majority of the bacterial communities and Lactobacillus amylolyticus being identified as the most prevalent and abundant species. The yeast communities were mainly composed of fermentative taxa, including Pichia kudriavzevii and Kluyveromyces marxianus. Lactobacillus amylolyticus and P. kudriavzevii were also the most frequently isolated species among bacteria and yeasts, respectively. These results highlighted a strong adaptation of the microbial isolates to starch-rich cereal substrates and underscored the central role of these microorganisms in wheat bran fermentation for bors production. Whereas the sequencing-based analyses showed no significant differences in overall diversity between the commercial and homemade bors samples, the cultivation-based results indicated a higher bacterial richness in the commercial products. Notably, the culture-dependent method captured substantially fewer taxa, highlighting the complementary nature of the two approaches. Of a total of 101 bacterial strains (88 LAB and 13 acetic acid bacteria) isolated, many exhibited rapid growth and strong acidification capacity, reaching pH values below 4.5 within 12 h. A functional screening revealed that 21 % of these strains displayed -amylase activity, 65 % phytase activity, and 50 % {beta}-glucosidase activity, highlighting their capacity to metabolize cereal substrates and enhance the nutrient availability of bors. All strains showed antibacterial activity against at least one indicator bacterium tested, with a universal inhibition of Listeria monocytogenes. Overall, Romanian bors harbored a lactic acid bacteria-dominated core microbiome with a significant functional diversity. These findings underscored its potential as a rich source of functional and technologically important strains for application in starter and protective culture development.

Volatile organic compounds (VOCs) define the distinctive aroma of cannabis and critically influence consumer preference, cultivar authentication, and breeding programs. However, systematic characterization of VOC diversity across commercial drug-type cultivars remains limited. This study presents a comprehensive volatilomics-based phenotypic characterization of 165 commercial drug-type cannabis accessions using gas chromatography with flame ionization detection and mass spectrometry (GC-FID/MS). We identified 61 high-confidence VOCs assigned to three biosynthetic classes: terpenoids (n = 45), fatty acid-derived volatiles (n = 12) and amino acid-derived volatiles (n = 4), resolved into 12 subclasses. Analysis of variance revealed highly significant among-accession differences for all compounds (p < 0.001; 2 = 0.67-0.97), with repeatability estimates averaging 0.81 (range 0.50-0.95). Unsupervised clustering partitioned accessions into three distinct chemotypes (n = 90, 53, and 22), supported by principal component and t-SNE analyses. Machine learning-based feature selection identified a consensus panel of 12 discriminative compounds (camphene, -fenchene, sabinene, -terpinene, ({+/-})-limonene, -humulene, linalool, endo-fenchol, {Delta}3-carene, -thujene, {gamma}-terpinene and -phellandrene) that recovered the chemotype assignment of 32 of 33 held-out accessions. Estimated odor-activity screening ranked prenylthiol, -pinene, ({+/-})-limonene, linalool and myrcene highest among the odor-active compounds. All three chemotypes shared a prenylthiol-dominated core (67-77% of summed OAV) and were distinguished by the extent and nature of terpenoid modulation of that core: minimally modulated (Cluster ZERO), citrus-floral modulated (Cluster ONE) and pine-terpenic modulated (Cluster TWO). These findings indicate that volatile diversity in this panel can be summarized by three reproducible chemotypes, providing a quantitative basis for accession characterization and a foundation for future breeding and quality-assessment studies.

Micronutrient deficiencies are prevalent in Madagascar, where diets rely heavily on starchy staples and access to animal-source foods is limited. Small dried fish (SDF) are widely available, yet their nutritional value and health risks remain poorly documented. We combined market surveys, taxonomic identification, and micronutrient and heavy metal analyses of nine SDF types collected along National Road 7. The samples encompassed 33 fish families, were dominated by small pelagic species (Clupeidae and Engraulidae), and were appreciated by consumers. A daily portion (5 g for infants; 10 g for young children and women of childbearing age) contributed substantially to Recommended Nutrient Intakes (RNIs). Across samples and groups, SDF were rich (>30% of RNI) in selenium and, for infants and young children, in calcium. All samples were a source of (>15% of RNI), or rich in, phosphorus, whereas iron contributions were more variable but often substantial. Several samples exceeded 100% of RNIs for selenium, calcium, iron, or manganese in infants and young children, and some were also sources of magnesium and, less frequently, zinc. Vitamin A was absent from sun-dried samples but detected in a smoked freshwater type. Heavy metal concentrations varied markedly, and portions of several types led to estimated exposures to inorganic arsenic or cadmium exceeding reference values, whereas freshwater species and some pelagic types showed a more favorable nutrition-risk balance. Overall, SDF are affordable, nutrient-dense foods with strong potential to alleviate micronutrient deficiencies in Madagascar, while highlighting the need for type-specific guidance to balance nutritional benefits and contamination risks.

Type 2 diabetes (T2D) is a prevalent metabolic disorder affecting millions worldwide, characterized by insulin resistance and impaired glucose homeostasis. While mammalian models are widely used, Drosophila melanogaster provides a powerful alternative due to its conserved insulin signaling pathways, genetic tractability, and suitability for high throughput studies. In addition to glucose dysregulation, lipid metabolism plays a crucial role in T2D pathophysiology, as alterations in lipid composition contribute to insulin resistance and metabolic dysfunction. Lipidomic studies have emerged as an essential approach to identify metabolic signatures and potential biomarkers for disease progression and therapeutic targeting. In this study, T2D like model was established by inducing insulin resistance through knockdown of the insulin receptor in brain insulin-producing cells using the dilp2-Gal4>UAS-InRRNAi system. This genetic manipulation resulted in significant metabolic dysregulation, including elevated glucose, trehalose, and triacylglyceride levels, along with increased oxidative stress indicators. Additionally, mRNA expression analysis of key insulin signaling components, including insulin receptor substrate 1, dilp2, dilp3, dilp5, and phosphorylated Akt, further validated the model. To further investigate metabolic alterations, Lipid profiling was performed using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) in non targeted LC-MS-based metabolomics approach to identify lipid biomarkers associated with T2D. Multivariate statistical analyses, including PCA and PLS-DA, revealed distinct lipid signatures between wild-type and T2D flies. Notably, specific phosphatidylglycerol species PG 34:0, PG 34:4, PA 38:3, PIP 38:1, PIP2 38:6, and LPS 24:0 demonstrated an area under the curve (AUC) of 1, indicating their strong reliability as lipid biomarkers for T2D diagnosis.