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Food Research International

○ Elsevier BV

Preprints posted in the last 90 days, ranked by how well they match Food Research International's content profile, based on 11 papers previously published here. The average preprint has a 0.01% match score for this journal, so anything above that is already an above-average fit.

1
Impact of Adipogenic Differentiation on the Nutrition and Sensory Profile of Cultivated Pork Fat Tissue

Lew, E. T.; Sugama, N.; Magrath, S. S.; Frost, S. C.; Kaplan, D. L.

2026-09-18 bioengineering 10.64898/2026.09.16.749166 medRxiv
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Adipose tissue is a principal source of texture, flavor, and aroma in meat products. Higher fat content corresponds to higher juiciness, tenderness and overall palatability of meat products, making it a key component of cultivated meat. Our previous study investigated the aromatic characteristics of cultivated pork fat tissue, finding comparable consumer panel hedonic scores between conventional livestock derived and cultivated pork fats. Further research was pursued to characterize if adipogenic differentiation of the cells is required to enhance the flavor of cultivated fat. While differentiation of cultivated cells results in tissues that more closely resemble the morphology of livestock-derived tissue, the process substantially increases production timelines and material costs associated with cultivated meat production. As a result, we observed changes in organoleptic and nutritional properties of cultivated pork fat cells associated with differentiation of porcine dedifferentiated fat (PDFAT) cells. We assessed the effect of varying degrees of adipogenesis with soybean oil supplementation on fat content, composition, and volatile compound formation. Notably, we observed the concentrations of 2-pentylfuran, 2-heptanone, hexanal and (E)-2-octenal increased compared to the undifferentiated preadipocytes. These compounds, along with other ketones, aldehydes, alcohol and lactone molecules, contribute to fruity, herbal, and fatty aromas. Additionally, these results were achieved without requiring the maximum differentiation time frame of 12 days in culture. These findings emphasize the importance of adipocyte differentiation in cultivated fat production and highlight its potential as a strategy for optimizing both flavor and nutritional attributes of cultivated meat.

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Open-source benchmarking of dairy and dairy-free products

Koosis, A. O.; Cotto, C.; Kuhl, E.

2026-08-19 bioengineering 10.64898/2026.08.14.744929 medRxiv
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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.

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Decoding the microbiota of bors: multifunctional potential of a traditional Romanian beverage fermentation

Grosu-Tudor, S.-S.; Meyer, A.; Angelescu, I. R.; Ionetic, E.-C.; Chirea, E.-T.; Bokulich, N.; Weckx, S.; De Vuyst, L.; Zamfir, M.

2026-08-13 microbiology 10.64898/2026.08.13.744363 medRxiv
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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.

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Pasting behaviour of Australian native grain flours: effects of particle size, flour blending, and temperature

Abedi, F.; Keitel, C.; Khoddami, A.; Roberts, T. H.

2026-09-30 plant biology 10.64898/2026.09.29.755466 medRxiv
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Knowledge of the influence of particle size, blending, and processing temperature on flour functionality is critical for the utilisation of emerging grains. This study investigated the flour pasting behaviour of four Australian native grains - Button Grass, Curly Mitchell Grass, Native Millet, and Weeping Grass - in relation to particle size, wheat flour substitution at various levels (0-30%), and elevated RVA temperatures (95-140 degrees Celcius). The pasting properties of flour fractions (fine, medium, and wholegrain) were analysed using a Rapid Visco Analyser, and water absorption capacity was also determined. Particle size effects were species-specific. Native Millet and wheat had higher viscosity in the finer fractions, whereas Weeping Grass exhibited maximum viscosity in the medium fraction. Button Grass and Curly Mitchell Grass showed limited sensitivity to particle size, indicating a stronger influence of intrinsic structure. Increasing substitution with native flours generally reduced viscosity, except for Weeping Grass, which was less affected and maintained stable viscosity. High-temperature rapid visco analysis revealed distinct thermal responses: most grains showed reduced final viscosity at elevated temperatures, whereas Weeping Grass maintained relatively high final viscosity across most temperatures. These findings demonstrate the diverse functional behaviour of Australian native grains and provide insights into their potential application in gluten-free and composite flour systems.

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Postprandial amino acid profiles and gastric behavior after consumption of milk and plant-based alternatives: a randomized cross-over study in healthy males

Leenders, L. M.; Casagranda, S.; Balvers, M. G. J.; de Gier, G.; Lambers, T. T.; Smeets, P. A. M.

2026-09-25 nutrition 10.64898/2026.09.23.26363226 medRxiv
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Background: There is a growing availability of plant-based alternatives for bovine milk. However, plant-based drinks differ e.g. in protein quantity, protein quality and carbohydrate composition. Knowledge on the impact this has on digestion-related outcomes such as postprandial amino acid (AA) availability and gastric behavior is still limited. Additionally, data on milk alternatives with higher overall protein concentration are lacking. Objective: We aimed to compare the differences in postprandial AA profiles, glycemic response, and gastric behavior between bovine milk and plant-based drinks with or without added protein using representative commercial drinks. Methods: 12 healthy males participated in a randomized crossover study with three treatments. After an overnight fast, participants drank 750 ml of semi-skimmed milk (Milk), a standard oat-based drink (OBD) or an oat-based drink enriched with pea protein (OBD+Pea). Blood samples were taken for measurement of plasma AA, glucose, insulin and FGF21 over 5 hours. Gastric content changes were assessed using magnetic resonance imaging scans for Milk and OBD+Pea over 2 hours. Results: Compared with Milk, both oat-based drinks elicited lower 5-h total AA iAUC (OBD -169706; OBD+pea -139552 microM*min; both P<0.001) and EAA iAUC (OBD -91748; OBD+pea -78199; both P<0.001), with no differences between oat-based drinks. Gastric content volume was lower for OBD+Pea versus Milk from 60-120 min (range -48 - -84 ml, all P<0.001). Compared to Milk, OBD and OBD+Pea increased glucose more from 15-45 min (range 0.86 - 1.73 mmol/l, all P<0.001) and insulin from 30-60 min (1.84 - 2.14-fold, all P<0.001). Conclusions: Protein enrichment in plant-based milk alternatives can be a valuable strategy to increase protein content and improve amino acid composition. However, such products still show faster gastric emptying, lower postprandial essential amino acid and higher glycemic and insulinemic responses compared to bovine milk because of their different carbohydrate and protein content and quality.

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Probiotic-Directed Fermentation Reprograms the Metabolic Profile of a Traditional Mongolian Whole-Wheat Diet and Modulates Escherichia coli-Induced Gut Microbiota Dysbiosis

duleng, E.; Ling, Q.; Bao, J.; Gaga, S.; gexi, T.; dien, N.; dan, S.; ruhan, A.; Bai, Y.; A, L.; Gong, C.; batu, B.; Ni, S.; Ping, W.

2026-08-11 microbiology 10.64898/2026.08.08.743650 medRxiv
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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.

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Development of a comprehensive database of markers of ultra-processing to identify ultra-processed food in the UK according to the Nova classification

Dicken, S. J.; van Tulleken, D.

2026-09-25 nutrition 10.64898/2026.09.24.26363859 medRxiv
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Growing evidence links ultra-processed food intake according to the Nova classification with poor health, yet no comprehensive database of additives and food substances of non-culinary use that are markers of ultra-processing exists for the UK. Building the database involved seven steps: (1) obtaining all approved additives in the UK, (2) retrieving all of their technical functions, (3) coding each technical function as cosmetic (marker of ultra-processing) or non-cosmetic (not a marker of ultra-processing), (4) adding 'flavour extracts', 'natural flavourings', and 'flavourings', (5) excluding additives as ultra-processing markers if they served fortification purposes, or were a herb/spice, and (6) adding food substances of non-culinary use defined as markers of ultra-processing in the Nova classification. Lastly, (7) two UPF definitions were constructed based on the ambiguity/overlap of additive technical functions from previous approaches. The database is freely accessible for researchers to improve consistency in using the Nova classification in the UK.

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Electro-Fermentation of Grape Must via Candida tropicalis SY005: Accelerating Kinetics, Modulating Biochemical Pathway, and Improving Bio-active Content

Sharma, S.; Gautam, S.; Gaidher, M.

2026-07-25 microbiology 10.64898/2026.07.25.740694 medRxiv
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This study investigates electro-fermentation candida Tropicalis SY005 to address fermentation kinetics limitation during grape must fermentation. In comparison with non-stimulated control sample, EF substantially enhanced sugar depletion, TSS drop by day 3 and generated a strongly reduced state (ORP -100 to -143mV). The oxidation-reduction shift enhanced cellular NAD+ regeneration, reducing total fermentation duration from 264 h to 72 h. GC-MS analysis showed pronounced major characteristic volatile compound confirming substantial metabolic pathway shifts in flavor of glycolytic flux. Moreover moderate electric field promoted cellular membrane electropermeabilization substantially promoting bioactive extraction.

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Characterization of Stress Tolerance and Probiotic Traits of Streptococcus salivarius Strains Isolated from Wildflower Honey

Lee, W.; Oh, Y.-S.; Ock, C.-S.; Oh, Y.-E.; Kim, H.-y.; Do, H.-K.; Hwang, C.-W.

2026-09-28 microbiology 10.64898/2026.09.27.754824 medRxiv
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Aims Honey represents a distinct microbial habitat, imposing physicochemical stresses on its resident microbiota while also serving as a potential source of beneficial microorganisms. This study aimed to isolate lactic acid bacteria (LAB) from Korean wildflower honey and characterize their gastrointestinal (GI) and physiological stress tolerance, adhesion-related properties, functional characteristics, and safety to evaluate their potential as probiotic candidates. Methods and Results Two Streptococcus salivarius strains, DOW-5A and DOW-10, were isolated from Korean wildflower honey and identified by phenotypic and molecular analyses. Both strains survived in sequential exposure to simulated GI conditions and showed tolerance to physiological stresses. DOW-5A maintained higher viability during the simulated gastric phase, whereas DOW-10 showed an initial decrease in viable counts followed by an increase during the simulated intestinal phase. The two strains also exhibited various probiotic characteristics, including mucin adhesion and antioxidant activity. Atypical, crystalline colony morphologies associated with EPS production were observed when sucrose was supplied as a carbon source. Both strains formed biofilms, while DOW-5A showed a marked increase in biofilm formation upon sucrose supplementation. Neither strain showed gelatinase or hemolytic activity or significant cytotoxicity toward Caco-2 cells. Impact Statement In this study, we isolated two honey-derived S. salivarius strains with high probiotic potential. Our findings highlight wildflower honey as a potential reservoir of LAB with traits relevant to survival under GI and physiological stresses, as well as various probiotic traits.

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Genome-Resolved Characterization of Stage-Associated Microbial Composition and Functional Potential During Spontaneous Ugba Fermentation

DIKE, K.; Ojukwu, C. C.; Oladele, F. T.; Ezeh, C. F.; Nwachukwu, M. I.

2026-09-26 microbiology 10.64898/2026.09.26.754590 medRxiv
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Ugba, a traditional alkaline-fermented condiment produced from African oil bean seeds (Pentaclethra macrophylla Benth.), remains poorly characterised at genome resolution. This study characterised a single spontaneous ugba fermentation using genome-resolved shotgun metagenomics across Early (0-24 h), Mid (48-96 h), and Late (120-144 h) composite stages. Thirty metagenome-assembled genomes (MAGs) were recovered and grouped according to their temporal abundance patterns. The Early-stage retained MAG population was dominated by Bacteroidota-associated MAGs, which exhibited the highest carbohydrate-active enzyme density and broad glycoside hydrolase repertoires, consistent with substantial genome-encoded potential for utilisation of the plant-derived seed matrix. The Mid stage showed marked restructuring of MAG composition, while Core and Late-associated MAGs increased towards fermentation maturation. Functional differentiation was also evident among Corynebacterium MAGs: C. nuruki encoded a complete urease system, whereas C. phoceense_A encoded glutamate dehydrogenase and nitrate-reduction genes. These stage-associated changes coincided with progressive alkalinisation from pH 7.15 to 8.01. Together, the results show that temporal restructuring of reconstructed microbial populations was accompanied by differentiation in genome-encoded functional potential. This study provides a genome-resolved characterisation of microbial composition and functional potential during spontaneous ugba fermentation and expands current understanding of the microbial organisation of this traditional African fermented food.

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Curcumin Enhances Alcoholic Fermentation in Yeast by Promoting Osmotic Stress Adaptation

Nakase, Y.; Kim, S.; Sakaue, K.; Sugimoto, Y.; Seike, T.; Watanabe, M.; Tohge, T.; Morozumi, Y.; Watanabeb, D.

2026-09-13 microbiology 10.64898/2026.09.10.750560 medRxiv
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Spices contain diverse plant secondary metabolites with antimicrobial and antioxidant activities that can influence plant-microbe interactions. Among these metabolites, curcumin and piperine are major bioactive constituents of turmeric and black pepper, respectively. In this study, we demonstrate that curcumin and piperine enhance alcoholic fermentation in the budding yeast Saccharomyces cerevisiae. Curcumin significantly promoted alcoholic fermentation under high-glucose conditions (20-40% glucose), which impose substantial osmotic stress on yeast cells. Under these conditions, curcumin enhanced yeast growth, improved osmotic stress tolerance, and increased resistance to cell wall digestion, suggesting enhanced stress adaptation during fermentation. Furthermore, deletion of HOG1, which encodes the central MAP kinase of the High-Osmolarity Glycerol (HOG) pathway responsible for the osmotic stress response, abolished the fermentation-enhancing effect of curcumin, indicating that Hog1 function is required for this effect. Consistent with this result, transcriptomic and RT-qPCR analyses revealed increased expression of stress adaptation-related genes, including known HOG pathway-responsive genes. In contrast, the expression of genes involved in amino acid catabolism was decreased at 12 h after the initiation of fermentation, while metabolomic analysis revealed increased intracellular levels of amino acids, including glutamate and asparagine, at 24 h. These coordinated transcriptional and metabolic changes are consistent with reduced amino acid catabolism and a shift toward a metabolic state favorable for sustained fermentation. Together, these findings suggest that curcumin enhances alcoholic fermentation under osmotic stress by promoting stress adaptation that supports yeast growth and sustained fermentation.

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Personalized nutrition products in a hospital setting: A prospective proof-of-concept study of acceptability in hospitalized patients and staff.

Melchers, M.; van der Haar, S.; Hoevenaars, F. P.; Wopereis, S.; Ummels, M.; van Bokhoven, K.; Hoppenbrouwers, M.; Rijfers, A.; Menkveld, M.; Noort, M. W.; van Zanten, A. R.; Kouw, I. W.

2026-10-05 nutrition 10.64898/2026.10.01.26364528 medRxiv
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Background & Aims Personalized nutrition has the potential to improve nutritional intake and clinical outcomes in hospitalized patients by adapting to individual dietary needs and preferences. Despite its promise, no personalized nutrition product (PNP) for hospitalized patients is currently available. This study aimed to assess the acceptability of PNPs among hospitalized patients and hospital staff. Methods A prospective, ten-days, proof-of-concept study was conducted among in-hospital adult patients ([&ge;]18y). Study patients received a freshly 3D printed PNP, customized to match individual taste preferences and health profiles. Patients could reorder the product daily if they remained hospitalized >24h after consumption. Consumption was assessed by weighing the PNP before and after consumption. Acceptability, sensory feedback, and overall experience were evaluated using questionnaires. Hospital staff also evaluated sensory feedback and acceptability. Factors influencing consumption and liking among patients were evaluated by testing within-group differences. Correlations between consumption rates, liking and patient characteristics were assessed. Results Eighty-seven patients received 137 PNPs during the study period. The average overall liking score was 5.7 ({+/-}2.1) on a 1-9 scale, and median consumption rate was 51% [Interquartile Range 19-100]. Consumption rates positively correlated with overall liking (R2 = 0.50, p < 0.001). Patients' willingness to reorder was 3.5 ({+/-}2.3) on a 1-7 scale, and reorder rate 63%. The perceived value of a PNP service in a hospital setting (scale 1-7) was scored 5.3 ({+/-}1.7) by patients and 4.5 ({+/-}1.1) by staff. Conclusions PNPs were evaluated as moderately positive in terms of acceptability and value by both patients and staff. However, further optimization of the product is needed before evaluating clinical efficacy.

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Isolation and Characterization of Bacteriocin-Producing Lactic Acid Bacteria from Cheese and Functional Evaluation of Their Synthesized Bioactive Peptides

Anumudu, C. K.; Miri, T.; Onyeaka, H.

2026-08-18 microbiology 10.64898/2026.08.14.744830 medRxiv
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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.

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β-lactoglobulin a new whey: Computational redesign improves stability and nutritional composition

Greis, M.; Castet, U.; Berlin, E.; Klangby, S.; Bancerz-Aleksiejczuk, O.; Vilaplana, F.; Keppler, J. K.; Hudson, E. P.

2026-08-18 bioengineering 10.64898/2026.08.17.745312 medRxiv
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Protein engineering and precision fermentation provide an opportunity to increase the value of food proteins by improving their solubility, stability, functionality, or nutritional composition. Here, we use {beta}-lactoglobulin ({beta}LG) as a model protein to investigate how state-of-the-art computational protein design approaches affect these properties. First, the deep learning-based design tool ProteinMPNN was used to alter up to 20% of {beta}LG residues for increased stability. Second, the physics-based modeling platform PyRosetta was used to find positions in {beta}LG accommodating increased branched-chain amino acid (BCAA) content and up to 10 residues were simultaneously exchanged. Experimental characterisation of ProteinMPNN and stabilised BCAA-enriched variants showed similar secondary structure and oligomeric state as native {beta}LG. ProteinMPNN variants gave increased titers and increased thermal stability up to 15 {degrees}C, and this correlated with changes in the rate of surface pressure in droplet tensiometry. Stabilized BCAA-enriched mutants had altered acid solubility. Correlations between computationally derived biophysical metrics and experimental properties are presented and suggest some predictive power for surface hydrophobicity on protein yield.

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Enabling predictive modeling of molecular connections between fermented foods and human inflammation through a paired dataset of cell-based models and multi-omics approaches

McDaniel, E. A.; Schertler, M.; Edillor, C.; Dutton, R. J.

2026-09-23 microbiology 10.64898/2026.09.22.753573 medRxiv
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Fermented foods are recognized for their rich microbial diversity and bioactive metabolites, which have recently been linked to anti-inflammatory effects and increased gut microbiome diversity. Despite extensive research on fermented foods including large-scale metagenomic surveys and metabolite characterization, a comprehensive mechanistic understanding of how diverse fermented foods, their microbes, and resulting metabolites interact with human biological pathways remains limited. Here, we systematically profiled over 100 commercially available fermented foods for their potential to prevent inflammation using a human cell-based model. We then generated bulk RNA sequencing of the human cells under these treatment conditions, along with metagenomic sequencing and metabolomics of the fermented foods used in the assays. By generating sample-matched multi-omics data, our work aims to lay the scientific groundwork that will enable scientists to generate predictive models and testable hypotheses about the molecular mechanisms underlying the anti-inflammatory effects of fermented foods. This open-source resource comprising all raw data and parsed intermediary files allows the community to begin elucidating the molecular mechanisms between fermented foods and human biology, paving the way for new, scientifically informed strategies for the user of fermented foods as functional foods.

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Microencapsulation of Nisin in Polyelectric complexes of alginate-chitosan for extended antimicrobial activity

Anumudu, C. K.; Miri, T.; Onyeaka, H.

2026-08-17 microbiology 10.64898/2026.08.14.744846 medRxiv
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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.

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Antibacterial Activity Potential of Lactic Acid Bacteria (LAB) Isolates from Palm Sap (Arenga pinnata) from the Wawo Plantation, Tomohon City, North Sulawesi

Pinaria, Y. W.; Pangkerego, N. P.; Kumolontang, G.

2026-08-24 microbiology 10.64898/2026.08.22.746455 medRxiv
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"Lactic acid bacteria (LAB) are one of the dominant groups of bacteria in the palm sap (Arenga pinnata) microbiome. Previous research in the sago palm sap production centers of Tomohon City (Kayawu, Pinaras, and Lahendong) has successfully identified various LAB species, including Lactobacillus casei, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus buchneri, Leuconostoc mesenteroides, and Leuconostoc sp. This study aims to identify LAB species in sago palm sap from a new location, namely the Wawo Plantation in Tomohon, and to evaluate their potential as natural antibacterial agents. Through 16S rDNA gene sequencing analysis, the isolates obtained were identified as belonging to the newly described genera Lacticaseibacillus and Lactiplantibacillus. Four promising isolates Lactiplantibacillus fabifermentans A1.4, Lacticaseibacillus casei B1.5, Lacticaseibacillus paracasei B1.6, and Lacticaseibacillus paracasei B3.5 were tested for their inhibitory activity against the enteric pathogens Salmonella sp. and Escherichia coli using the well diffusion method. The results showed that all isolates exhibited a strong spectrum of pathogen inhibition. The highest inhibitory activity against Salmonella sp. was demonstrated by the L. paracasei B1.6 isolate, with an inhibition zone of 21.25 mm, while optimal inhibition against E. coli was achieved by L. casei B1.5 at 11.0 mm. These findings confirm that the local BAL strain from Tomohon palm sap has great potential for large-scale development as a biopreservative in the food industry and as a functional probiotic agent"

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Challenges of Dry Sanitization to Control Salmonella Dry Surface Biofilms

Vaz, V.; Finger, J.; Pereira, R. F.; Santiago Silva, E.; Pimentel Maia, R.; Maillard, J.-Y.; Nascimento, M.

2026-08-06 microbiology 10.64898/2026.08.06.743265 medRxiv
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Salmonella is a pathogen linked to foodborne outbreaks, including low-moisture foods. Its ability to resist desiccation can contribute to the formation of dry surface biofilms (DSB). This study evaluated the impact of 3 DSB formation protocols (P1=48-h hydrated phase/48-h dry phase, P2=24-h/120-h and P3=8-h/48-h) on the resistance of Salmonella DSB to 70% alcohol, a commercial product (based on 0.015% quaternary ammonium and 25% isopropyl alcohol), gaseous ozone (45 ppm), hot air (90 {degrees}C) and UV-C light (254 nm). The type of DSB protocol impacted the efficacy of the sanitizers (p < 0.05). The biofilm with the shortest hydration phase showed the greatest susceptibility; three out of the five sanitizers evaluated (70% alcohol, commercial product, and UV-C) promoted significant reductions in P3, with counts below the detection limit (0.8 log CFU/cm{superscript 2}) after 5 to 15 min exposure. Regarding protocols P1 and P2, in general, the best performance was from UV-C, especially against DSB on polypropylene, where it achieved reductions of 1.3 log CFU/cm{superscript 2} for P1 and 2.9 log CFU/cm{superscript 2} for P2 after 15 to 30 min of exposure. In contrast, hot air and ozone showed less effectiveness, with reductions [&le;]1.2 log CFU/cm{superscript 2}. In most scenarios, confocal microscopy images corroborated the plate count results (log CFU/cm{superscript 2}). In summary, our data indicates limited action of dry sanitizers on Salmonella DSB, requiring validation and optimization of sanitization processes to ensure the microbiological safety of low-moisture products.

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Lipogenic gene expression and substrate sensitivity in the bovine mammary gland shape milk fat composition

Tzirkel-Hancock, N.; Rakover, M.; Tadmor-Levi, R.; Argov-Argaman, N.

2026-09-01 cell biology 10.64898/2026.08.31.748247 medRxiv
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2.1%
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Milk fat is produced by mammary epithelial cells (MEC) through a conserved mechanism shared among all fat-producing cells across biological kingdoms. Although highly conserved, different tissues and organisms produce distinctive fat compositions. Notably, ruminant milk fat is characterized by enrichment in short and medium chain fatty acids. We hypothesized that this unique profile is driven by MEC-specific metabolic characteristics related to their response to lipogenic substrates. To study this, we compared bovine MEC and udder-derived fibroblasts in terms of their lipogenic capacity and fatty acid composition when exposed to lipogenic building blocks. When exposed to acetate, MEC showed coordinated upregulation of acyl-CoA short-chain synthetase 1 (ACSS1) and diacylglycerol transferase (DGAT), while expression of acyl-CoA synthetase long-chain 1 (ACSL1) decreased. Medium chain fatty acids were also elevated in acetate-treated MEC and not in fibroblasts. The role of ACSS1 in the production of medium chain fatty acids in MEC was confirmed by knockdown experiments. Metabolomics analysis showed that in MEC acetate treatment triggered a broad metabolic response, primarily amino acids catabolism, energy and polar lipid metabolism. Collectively, these findings demonstrate effective utilization of acetate for de novo fatty acid synthesis in MEC with preferred tendency to produce medium chain fatty acids.

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Effect of Mushroom-Bacteria Co-culture on Mushroom Growth and Antimicrobial Properties

Wang, E.; Cavanaugh, N. T.; He, Y.; Chai, Y.

2026-08-31 microbiology 10.64898/2026.08.30.747672 medRxiv
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1.8%
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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.