Search Results (1,354)

The growing demand for plant-based functional foods has encouraged the development of non-dairy fermented beverages able to deliver viable microorganisms and improve the technological and nutritional value of plant matrices. This study aimed to develop a prototype fermented peanut beverage using Lactiplantibacillus plantarum strains and to evaluate their technological performance, stability, and functional potential. Two plant derived strains, L. plantarum F1B and LpAv, and the commercial probiotic L. plantarum 299v were assessed in animal-free culture media and used to ferment peanut extract supplemented with sucrose and yeast extract. Fermented beverages were characterized through microbiological, physicochemical, rheological, peptide profile, refrigerated storage, and simulated gastrointestinal digestion. The selected formulation allowed all strains to reach pH ≤ 4.5 after 6.5 h, with LAB counts of 8.6–9.0 log CFU/mL. Lactic acid was the main fermentation product, and oxalic acid levels were significantly reduced. Fermentation increased peptide signals and improved rheological behavior, generating pseudoplastic beverages with higher viscosity than the control. LAB counts remained ≥8 log CFU/mL after 28 days at 4 °C, although gastrointestinal resistance after storage was strain-dependent. Taken together, L. plantarum strains showed promising technological and functional properties for developing fermented peanut-based beverages. The nutritional properties of peanut, together with the fact that it supports the growth and survival of LAB in a fermented beverage, positions peanuts as a candidate which deserves further studies in the space of fermented functional foods. Full article

The growing global demand for ethical, resource-efficient protein sources has renewed serious interest in cultured meat as a viable alternative to conventional livestock production. Two revolutionary biotechnological systems, induced pluripotent stem cell (iPSC) reprogramming and CRISPR-Cas9-mediated genome editing, when combined, offer unparalleled accuracy and scalability for the biofabrication of avian flesh. In this review, we present a comprehensive pipeline that involves the ectopic expression of Yamanaka factors (Oct4, Sox2, Klf4, and c-Myc) to reprogram primary somatic cells derived from Gallus gallus into induced pluripotent stem cells (iPSCs). This process is subsequently followed by targeted genome editing to enhance myogenic potential, growth efficiency, nutritional composition, and disease resistance. iPSCs are cultivated in a xeno-free bioreactor following genome editing, and subsequently directed to develop into myoblasts and mature myotubes. Three-dimensional tissue biofabrication is realized by combining biomaterial scaffolds and perfusion bioreactor systems, structuring an authentic muscle tissue matrix. These engineering platforms enable precise control over microenvironmental parameters, including oxygenation and nutrient perfusion. The resulting biofabricated poultry product is compositionally optimized, free of antibiotic residues, and exhibits a significantly reduced environmental footprint than poultry that is grown in the traditional way. This all-in-one solution solves important problems in food security, animal welfare, land use efficiency, and greenhouse gas emissions while also setting up a scalable biomanufacturing framework for making proteins for the next generation. Full article

Background: Incorporating probiotic bacteria such as BC, a spore-forming probiotic bacterium known for its resistance, into extruded snacks has emerged as an innovative approach to developing functional foods. Little is known about how the addition influences metabolomic changes during storage. Methods: The mung bean snacks were produced by extrusion, followed by BC fortification. The control snack without seasoning and the two BC treatments (0.1% and 0.5% w/w) were stored at room temperature for 3 months. Culture-based methods and qPCR were performed for microbial analysis. Metabolites were extracted, and metabolite profiles were evaluated using an LC-MS-based approach. Results: 0.5% BC was the most stable in the snack, with viable counts of 7.10 log CFU/g. The PCA plot demonstrated distinct clustering patterns among groups, despite no significant difference by PERMANOVA. In negative mode, L-tyrosine showed higher accumulation in the 0.1 BC group, whereas Threonine, L-histidine, and fatty acid metabolites were highly abundant in the 0.5 BC treatment. The positive mode showed discriminant metabolites, with L-glutamic acid, Isoleucine, and L-phenylalanine enriched in the 0.5 BC treatment. Overall, BC altered citrate and phenylalanine metabolism, and each treatment affected aminoacyl-tRNA biosynthesis. Conclusions: These findings demonstrate the maintenance of culturable B. coagulans in fortified extruded mung bean snacks during storage and provide exploratory insights into metabolite changes associated with amino acid metabolism, lipid oxidation, and flavor development that may be relevant to product quality and storage stability. Full article

The intensification of the urban heat island effect poses a critical threat to the preservation and habitability of compact historic districts. The Alameda de Hércules in Seville exemplifies this vulnerability, where the intersection of heritage protection and extreme Mediterranean summers limits conventional climate adaptation. This study conducts a multi-temporal evaluation of the square’s climate resilience, spanning from its configuration prior to major 21st-century renovations to its current state and future projections, proposing future interventions. By integrating advanced microclimatic simulation and high-fidelity energy modeling, the research assesses a dual-function strategy: the improvement of the thermal environment while implementing non-intrusive photovoltaic pavements (PVPs) for energy generation. Environmental parameters, including air temperature, mean radiant temperature (MRT), and the universal thermal climate index (UTCI), were analyzed alongside the renewable energy potential of the site’s mobility infrastructure. Four heritage-sensitive interventions were tested: PV-integrated bicycle lanes, shading canopies, reflective pavement, and permeable paved grass. The results demonstrate that the canopies and paved grass zones can lower surface temperature up to 3.7–4.3 °C, reduce UTCI stress up to 2.3–3.0 °C, and decline MRT up to 10.6 °C. These values correspond to the maximum reductions achieved in specific zones. However, the PVP can locally increase surface temperature by about 4.7 °C and the reflective pavements increase MRT by around 10.4 °C, while generating an estimated annual energy yield of 174.19 MWh. The analysis under future climate projections suggests that these strategies remain equally effective under future scenarios. These findings confirm that PV-integrated urban surfaces offer a viable, reversible, and replicable approach to retrofitting historic public spaces, harmonizing climate-adaptive cooling with decentralized energy production without compromising the site’s cultural significance. Full article

This study investigated lactic fermentation of green buckwheat beverages formulated at 8% (A, AA) and 10% (B, BB) solids using single- and multiple-strain cultures of Lactiplantibacillus plantarum, Lactobacillus acidophilus, Lacticaseibacillus paracasei and Lacticaseibacillus rhamnosus. Fermentation at 37 °C rapidly reduced pH from slightly alkaline values (7.44–7.57) to approximately 4.2–4.5 within 3–8 h, while viable counts increased from near-zero to 8–9 log₁₀CFU mL⁻¹, confirming efficient lactic acid bacteria (LAB) proliferation in all substrates. A general trend was observed in the sugar consumption strategy of studied LAB: sucrose (after hydrolysis) and glucose were almost completely depleted within 6–8 h, fructose was consumed more slowly, and raffinose remained largely unchanged, with the 10% substrate mainly accelerating early sugar turnover without altering final cell densities or the qualitative utilisation pattern. Dry matter changed little during fermentation, whereas total phenolic content (TPC) and total tannin content (TTC) were strongly affected in a matrix- and strain-dependent manner. At 8% solids, fermentation promoted substantial TTC reduction and, for several cultures, a net decrease in extractable phenolics. In contrast, 10% formulations, particularly those inoculated with L. acidophilus and L. paracasei (alone or in combination), partially preserved or increased TPC while achieving more moderate tannin losses. Overall, green buckwheat proved to be a promising substrate for developing fermented beverages in which solids level and starter composition can be tuned to combine rapid acidification and high LAB viability with tailored sugar depletion and favourable modulation of phenolic and tannin fractions. Full article

Botrytis cinerea, the causal agent of gray mold disease, is a major phytopathogen responsible for substantial losses in horticultural crops. In this study, cultivation conditions for Bacillus velezensis R22 were optimized to maximize overall antifungal activity against B. cinerea. A Plackett–Burman design was used to identify medium components affecting antifungal activity in flask cultures, followed by response surface methodology based on a central composite design (CCD) to optimize sucrose concentration, temperature, and agitation speed in a stirred bioreactor. Maximum antifungal activity was obtained at 17.45 g/L initial sucrose, 31.8 °C, and 293 rpm. The biological relevance of the optimized culture was confirmed in a tomato infection model, in which gray mold severity was reduced by 85.3% relative to the untreated control and by 59.9% relative to the non-optimized R22 culture. The same CCD approach was subsequently applied to determine cultivation conditions that maximize the concentration of R22 viable cells. The optimal parameters for 24-h growth (35.46 g/L sucrose, 36.5 °C, and 454 rpm) differed markedly from those identified for maximal antifungal activity. When evaluated on uninfected tomato plants, cultures produced under conditions favoring higher cell density showed enhanced plant growth-promoting activity compared to the non-optimized culture. Mass spectrometric analysis of lipopeptide extracts revealed that the enhanced antifungal activity was accompanied by an increased abundance of long-chain homologs across all major lipopeptide families, particularly surfactins. Thus, our results indicate that maximizing overall antifungal activity may be of greater practical significance than optimization of the individual fungicidal agent. Full article

Elite swimmers are exposed to sustained high training loads, early-morning sessions, and restricted recovery opportunities, all of which may increase psychological strain and compromise sleep. This study examined the effects of an 8-week Mindfulness–Acceptance–Insight–Commitment (MAIC) program, embedded within routine high-load training, on athlete-specific psychological stress, subjective sleep quality, and mindfulness in elite swimmers. A randomized controlled mixed-methods design was used. Thirty elite swimmers from a provincial high-performance program in China were randomly assigned to an MAIC group or a usual-practice control group (n = 15 per group). Quantitative outcomes were assessed at baseline, post-intervention, and three-month follow-up using the Athlete Psychological Strain Questionnaire, salivary cortisol, the Pittsburgh Sleep Quality Index, and the Athlete Mindfulness Questionnaire. Semi-structured interviews were conducted with all athletes in the MAIC group after the intervention. Mixed-design ANOVAs revealed significant Group × Time interactions for athlete-specific psychological stress, salivary cortisol, sleep quality, and mindfulness. Compared with the control group, the MAIC group showed lower psychological strain and cortisol, better subjective sleep quality, and higher mindfulness at post-intervention. At follow-up, improvements in psychological stress and mindfulness remained evident relative to baseline, whereas lower salivary cortisol and more favorable self-reported sleep quality remained evident relative to the control group. Qualitative findings further showed that MAIC was experienced as feasible, low-burden, and readily integrated into the training context. Athletes described attentional resets, acceptance-based responses to discomfort, and brief post-session or pre-sleep practices as helpful for regulating cognitive reactivity and arousal. Overall, MAIC appears to be a culturally grounded and practically viable adjunct strategy for supporting psychological regulation and self-reported sleep quality in elite swimmers during demanding training periods. Full article

Dental caries is a biofilm-mediated dysbiotic disease characterized by ecological shifts within the oral microbiome and progressive demineralization of dental hard tissues. The microbiological composition of carious dentin and the impact of minimally invasive excavation techniques on residual microbial communities remain subjects of ongoing investigation due to methodological heterogeneity and inconsistencies among published studies. This narrative review aimed to summarize current evidence regarding the microbial ecology of carious dentin, compare culture-based and molecular methods for microbiological assessment, and evaluate the microbiological outcomes associated with contemporary approaches to managing minimally invasive caries. The relevant literature on dentinal caries microbiology, microbial detection methods, and excavation techniques was analyzed. The available evidence indicates that carious dentin contains a highly diverse polymicrobial community composed of acidogenic, aciduric, anaerobic, and proteolytic microorganisms. Culture-based methods primarily detect viable and cultivable taxa, whereas molecular approaches reveal substantially greater microbial diversity, including uncultivable and low-abundance species. Comparative studies demonstrate that minimally invasive excavation techniques significantly reduce microbial load but rarely achieve complete microbial elimination. The available evidence suggests that successful caries management is associated with a reduction in and ecological modulation of the residual microbiota within a sealed environment. The integration of culture-based and molecular findings provides a more comprehensive understanding of the microbiology of carious dentin and supports biologically oriented, minimally invasive strategies for caries management. Full article

Edible insects are increasingly recognized as a viable alternative protein source, offering a potentially sustainable approach to addressing global food security challenges. This narrative review critically examines the nutritional composition, environmental advantages, techno-functional attributes, and potential applications of insect-based proteins within human food systems. Edible insects are characterized by high protein content, favourable essential amino acid profiles, and appreciable levels of key micronutrients, rendering them nutritionally comparable to conventional livestock-derived proteins. Moreover, insect production systems generally require substantially lower inputs of land, water, and feed, resulting in comparatively lower greenhouse gas emissions and reduced overall environmental burden. Despite these advantages, broader adoption remains constrained by challenges related to regulatory heterogeneity, food safety concerns, and limited consumer acceptance. Overall, the available evidence suggests that edible insects can function as a nutritionally adequate and environmentally sustainable complementary protein source; however, significant variability in nutrient composition, limitations in standardized safety assessment, and socio-cultural barriers currently restrict their large-scale integration into mainstream food systems. In addition, inconsistencies in analytical methodologies and reliance on in vitro data further complicate cross-study comparisons and translational relevance. Future research should focus on standardization of rearing and processing conditions, harmonization of evaluation frameworks (e.g., protein quality indices), comprehensive safety assessments, and well-designed clinical studies to validate nutritional and functional benefits, alongside the development of effective strategies to improve consumer acceptance and support regulatory alignment across regions. Full article

The increasing generation of agro-industrial residues like nejayote (maize lime-cooking wastewater from the maize nixtamalization process) poses significant environmental challenges in Mexico due to its elevated chemical oxygen demand (COD) and organic load. This study evaluates the physical separation of nejayote via membranes and its use as a low-cost substrate for producing lipids and carotenoids using Rhodotorula glutinis. A batch culture followed by pulse-feeding achieved a COD removal efficiency of 53.6% (0.22 g COD/(L h)) and a biomass concentration of 3.72 ± 0.45 g COD/L within 48 h. The yeast demonstrated a high specific metabolic efficiency, yielding 0.457 g of lipids and 0.0049 g of carotenoids per gram of biomass, with an oleaginous fraction of 46.21% in dry weight. Experimental data calibrated a process model in SuperPro Designer, simulating full-scale processes treating 100, 1000, and 10,000 m³ of nejayote per batch, producing up to 2137.11 MT of lipids and 22.90 MT of carotenoids annually. A techno-economic analysis estimated the investment, operating costs, and financial indicators for all scenarios. Strategies like evaporation and reverse osmosis to concentrate nejayote significantly improved profitability by reducing equipment size. Additionally, a circular economy approach was modeled, recovering process water and nutrient-rich side streams. These findings confirm that integrated physical and biological treatment, coupled with resource recovery, transforms this particularly agro-industrial residue into a technically robust and economically viable biorefinery feedstock, aligning industrial production with sustainable waste management. Full article

Spoilage in vacuum-packaged chicken breast is driven by coupled microbial succession and physicochemical changes that cannot be adequately described by a single indicator. In this study, MALDI-TOF MS-based species-level identification of culturable isolates was integrated with microbiological counts (total viable count, lactic acid bacteria, yeasts and molds, and Enterobacteriaceae), physicochemical parameters (pH, water activity, CIE $L^{\ast }{a}^{\ast }{b}^{\ast }$, and total volatile basic nitrogen (TVB-N)), and sensory evaluation of odor, appearance/color, surface texture/slime and overall acceptability (trained panel, $n=8$) during 15 days of storage at 4 °C. Associations among variables were assessed using Spearman correlation analysis. MALDI-TOF MS identified 625 isolates belonging to 67 species across 19 families. The microbial community shifted from an initially diverse flora toward late-stage dominance by Latilactobacillus sakei, L. curvatus, Hafnia alvei, Serratia spp., Carnobacterium maltaromaticum and Brochothrix thermosphacta, while Candida zeylanoides persisted throughout storage. TVC exceeded 7 log CFU/g, and TVB-N increased from 10.65 to 23.20 mg N/100 g ($p<0.05$). TVB-N showed strong positive correlations with all microbial groups ($r_s\ge 0.90$, $p<0.01$) and with seven microbial families at the family level. Hafniaceae dominance coincided with a transient mid-storage decrease in pH, consistent with the deaminative activity of H. alvei. $b_{in}^{\ast }$ showed significant associations with four microbial families and with both microbial counts and TVB-N, supporting its value as a practical spoilage indicator. Sensory evaluation identified Day 13 as the rejection point, corresponding to TVC of 6.79 log CFU/g and TVB-N of 20.80 mg N/100 g, with simultaneous deterioration of odor and appearance, in contrast to the sequential pattern typically reported under aerobic conditions. To our knowledge, this is the first study to integrate time-resolved MALDI-TOF MS-based family-level profiling with physicochemical and sensory monitoring in vacuum-packaged chicken breast stored at 4 °C, offering a condition-specific framework for shelf-life assessment. Full article

Foodborne pathogens pose a major challenge for public health, food safety regulation, and industrial quality control. Effective surveillance, outbreak tracing, and early warning for foodborne microbial contamination require rapid, reliable detection methods. Conventional culture-based methods are still essential for regulatory confirmation since they recover viable isolates and support downstream verification. However, their long turnaround time, labor-intensive procedures, and limited throughput restrict their use in rapid screening and on-site testing. In recent years, immunological assays, nucleic acid amplification and recognition methods, biosensors, microfluidic systems, CRISPR-Cas platforms, mass spectrometry, sequencing technologies, and artificial intelligence-assisted analysis have expanded the detection toolbox. These methods improve speed, sensitivity, portability, and multiplexing capacity, but their performance still depends on food-matrix properties, sample pretreatment, and application conditions. This review compares representative methods in terms of analytical principle, sample pretreatment, sensitivity, specificity, assay time, viable-cell discrimination, field applicability, and standardization potential. In our opinion, culture-based methods are central for confirmation, while emerging technologies are better suited for rapid screening, integrated analysis, and point-of-need testing. Nevertheless, matrix interference, limited validation in naturally contaminated samples, insufficient viable/dead-cell discrimination, and weak cross-platform consistency remain key barriers to routine use. Full article

Background/Objectives: Probiotic feed additives are increasingly used in livestock production as antimicrobial-sparing tools, yet viable microbial products should not introduce clinically relevant antimicrobial resistance genes (ARGs) into the intestinal resistome. This study evaluated farm-animal probiotic products using an integrated phenotypic, metagenomic and mobilome-aware safety framework. Methods: Seven commercially available products intended for poultry, pigs, cattle or horses were assessed using product metadata, culture-based recovery, broth microdilution minimum inhibitory concentration (MIC) profiling and Illumina short-read sequencing as a screening-level resistome approach. Reads were quality controlled, assembled, screened using the Comprehensive Antibiotic Research Database (CARD)/Resistance Gene Identifier (RGI) workflow and interrogated for plasmid-, phage- and insertion sequence/mobile genetic element-associated genomic context. Results: MIC profiles were generated for viable bacterial isolates representing Enterococcus faecium, Pediococcus acidilactici, Pediococcus pentosaceus and Bacillus subtilis. One labelled Lactobacillus plantarum component was not recovered as viable culture, and one labelled P. acidilactici component was recorded as P. pentosaceus. Sequencing-based resistome screening identified 30 antimicrobial resistance (AMR)-associated CARD antibiotic-resistant organism (ARO) hits belonging to 13 determinants across six ARG-positive coded products, while one coded product had no retained CARD/RGI hit. Profiles were dominated by recurrent Enterococcus-associated background determinants, including aac(6′)-Ii, msrC and eatAv. Plasmid prediction was positive for five hits, whereas no iMGE- or phage-associated ARG context was detected. No vanA/vanB, mcr, optrA, poxtA, cfr, extended-spectrum β-lactamase (ESBL) or carbapenemase gene was detected. Conclusions: The investigated products did not show evidence of high-priority mobile ARG carriage. Nevertheless, AMR-associated determinants and occasional predicted mobile contexts support routine integrated MIC-sequencing-based resistome–mobilome assessment of veterinary probiotic products. Because short-read assemblies do not fully resolve plasmid architecture or transferability, mobile-context predictions should be considered screening-level indicators requiring confirmatory long-read or functional testing for higher-priority findings. Full article

Primary neuronal cultures from the brain are critical for investigating disease-specific cellular and molecular mechanisms in mouse models. Current methods for obtaining primary cultures require embryonic brains that are affected by embryonic lethality and genotypic characterization in severe disease models such as sickle cell disease (SCD). Furthermore, these neuronal cultures require about 14 days in vitro (DIVs) for neurite outgrowth to mature. We adapted and optimized a relatively simplified and reproducible method using brains from postnatal day 1 mouse pups for isolating and culturing hippocampal and cortical neurons. This approach produces viable neurons that attach, extend neurites, and express key synaptic markers by 7 DIV and also minimizes glial outgrowth. We successfully applied this approach to isolating and culturing hippocampal and cortical neurons from the brains of one-day-old (P1) pups of humanized transgenic homozygous BERK sickle cell and control mice. Morphological observations at 3, 7, and 14 DIVs demonstrated robust neuronal attachment, neurite outgrowth, and overall structural development in both male and female hippocampal and cortical neurons. Neurons in culture expressed key markers including neuronal nuclear protein (NeuN/Rbfox3), neurofilament 200 (NF200), microtubule-associated protein 2 (MAP2), vesicular glutamate transporter 1 (VGLUT1), postsynaptic density protein 95 (PSD 95), and glutamate N-methyl-D-aspartate receptor subunit 2B (GluN2B). Notably, male SCD hippocampal neurons evinced a higher density of PSD 95 puncta on dendritic spines compared to controls on 7 as well as 14 DIVs. Incubation of male hippocampal neurons in a sickle cell-like microenvironment with TNF-α and heme further increased the density of PSD 95 puncta and colocalization of GluN2B with PSD 95, supporting the utility of this culture system for examining disease-relevant structural and molecular responses. This optimized culture system provides a simplified and reproducible platform to investigate the mechanisms involving neuronal dysfunction in challenging mouse models of brain disorders. Full article

Efficient in vitro production of rabies virus is essential for vaccine development and quality control applications. High-density cultivation systems offer practical advantages for rabies virus production but also create culture conditions in which nutrient depletion, waste accumulation, and progressive deterioration of host-cell condition may limit infectious virus output. In this study, we evaluated the effects of sodium pyruvate supplementation on rabies virus CVS-11 production in Vero and BSR cells cultivated in a high-density macrocarrier-based tide-motion culture system under serum-containing and serum-free conditions, with complementary comparative observations in conventional monolayer cultures of BHK cells. Cultures were infected at a multiplicity of infection of 0.01, and infectious virus production was assessed over time, together with cell density, glucose consumption, and pH dynamics. Sodium pyruvate supplementation was associated with significantly higher infectious virus titers, delayed culture deterioration, prolonged maintenance of viable cell populations, and higher peak infectious titers in both Vero and BSR cultures. The highest infectious titers were observed under serum-free pyruvate-supplemented conditions, reaching 7.5 log₁₀ FFU/mL in Vero cells and 7.2 log₁₀ FFU/mL in BSR cells. Across the tested conditions, serum-free cultivation and pyruvate supplementation were both associated with significantly higher peak infectious titers. In contrast, exploratory correlation analysis based on condition-level summary values indicated an inverse association between minimum culture pH and peak infectious titer. Together, these findings show that sodium pyruvate supplementation can improve infectious rabies virus yield and prolong the productive phase in high-density macrocarrier-based cultures, supporting its use as a practical culture-modulation strategy for CVS-11 production in adherent cell systems. Full article