Pressed peanut cake (PPC) is a valuable plant-based protein source, but its complex matrix might lead to underestimation of aflatoxin B1 (AFB1). This study established a multi-technique diagnostic framework integrating UV-Vis, FTIR, LC-MS/MS, and SEM to systematically dissect matrix interference in different extraction solvents, ultimately correlating with AFB1 detection. It was found that, due to lipid interference, the use of nonpolar solvents during extraction not only significantly amplified detection errors but also severely compromised extraction efficiency, resulting in an analytical error exceeding 85%. Systematic profiling revealed that conventional methanol/water extraction introduced dual errors, attributable to either inefficient toxin extraction or excessive pigment co-extraction, leading to an AFB1 underestimation up to 50%. Acetonitrile/water was identified as the optimal extraction solvent, minimizing interferents and achieving consistent AFB1 recoveries >90%. This work not only provides a practically applicable detection protocol but also offers a clear mechanistic explanation for the observed errors.

Pressed peanut cake (PPC) is a valuable plant protein resource but is highly susceptible to aflatoxin B1 (AFB1) contamination. Its complex matrix can complicate the accurate analysis of AFB1. Digesting the samples in vitro prior to quantification resulted in a significant improvement in AFB1 detection value (25.81-39.40%) compared with untreated samples. Ultrafiltration analysis confirmed that conventional extraction solvents are susceptible to severe interference from the matrix. Molecular docking demonstrated that AFB1 can form complexes with the matrix. Protein hydrolysis leads to significantly higher detection values than starch or cellulose hydrolysis, a phenomenon attributed to the dominant role of protein in binding the toxin. Peanut protein entraps and locks AFB1 through synergistic hydrophobic and hydrogen bonding interactions, forming restricted matrix-bound AFB1. The study elucidated on the formation/ dissociation mechanism of matrix-bound AFB1 in PPC at the molecular level, emphasizing the underestimation of the actual exposure level of AFB1 in PPC.

Patulin, a toxic mycotoxin widely found in fruits and derived products, poses a serious threat to food safety. This study rapidly identified WaSDR, a novel short-chain dehydrogenase/reductase (SDR) from Wickerhamomyces anomalus XL1, using proteomic analysis and computational modeling. Molecular docking indicated that WaSDR had the strongest affinity for patulin with a binding energy of -6.9 kcal/mol among candidate proteins, and 100 ns molecular dynamics simulation verified the stability of the WaSDR-patulin complex. The WaSDR gene was cloned, expressed in Escherichia coli, and purified. At a concentration of 30 μg/mL, WaSDR completely degraded patulin into the isomers E-ascladiol and Z-ascladiol within 24 h. WaSDR showed optimal activity at pH 7.0-8.0 and 4-30 °C. In apple juice, WaSDR degraded 10 μg/mL of patulin within 48 h without affecting other quality parameters. This study provides an efficient strategy for identifying patulin-degrading enzymes and highlights WaSDR's potential for mycotoxin detoxification in the food industry.

Traditional Ras cheese from Egypt harbors complex microbial communities essential for flavor development and quality. This study combined metagenomic sequencing and culture-based isolation to identify key microbial species and explore their functional roles. From 11 Ras cheese samples collected in Alexandria and Cairo, five key species were identified, Enterococcus (E.) faecium, E. durans, Lacticaseibacillus rhamnosus, Limosilactobacillus fermentum, and Companilactobacillus versmoldensis, using causal inference and topological analysis of microbial interaction networks. Metagenome-assembled genomes revealed enrichment in amino acid, carbohydrate, and vitamin metabolism pathways linked to flavor formation. Whole-genome sequencing of 64 isolates enabled comparative analysis of E. faecium (n = 36) and E. durans (n = 28), revealing open pan-genomes, phylogenetic clades without geographic clustering, and significant intraspecific variation in genome size and GC content. Carbohydrate-active enzyme profiles showed clade-specific distributions, indicating functional diversification. Biosynthetic gene cluster prediction uncovered widespread potential for antimicrobial and quorum-sensing compound production. While four key species were successfully cultured, Companilactobacillus versmoldensis remained uncultured, highlighting limitations in current isolation methods. This integrative approach advances understanding of microbial ecology in traditional fermented dairy products and provides genomic resources for starter culture development.

Screening phage-resistant strains with desirable characteristics is an effective strategy to prevent phage contamination in industrial fermentation. In this study, a spontaneous phage-resistant mutant strain, IMAU10120-1, derived from Lactiplantibacillus plantarum (L. plantarum) IMAU10120, was obtained using secondary infection method. The biological characteristics and genomic mutations of the resistant strain were analyzed, and its impact on the flavor profile of fermented soymilk during storage were investigated. The results demonstrated that IMAU10120-1 exhibited strong phage resistance, stable passage inheritance, and no lysogenic properties. Comparative genomic analysis revealed multiple non-synonymous mutations in the resistant strain, affecting transcriptional regulation, energy coupling factors, and DNA-binding reactions. Notably, modifications in the shikimic acid metabolism pathway enhanced the strain's aroma-producing capacity, leading to the generation of more unique flavor compounds in fermented soymilk during storage. These findings provide valuable insights into the mechanisms of phage resistance and support the industrial application of phage-resistant L. plantarum strains.

This study investigated the efficiency of a novel polyvinyl alcohol-sodium alginate (PVA/SA) bead system for treating sesame butter wastewater (SBW) and concurrently producing bio-alkanes. The best two isolates were identified through 16S rRNA analysis and investigated as freely suspended individuals and in consortium treatments. The effect of bacterial immobilization on SBW detoxification was then investigated. Encapsulating Pseudomonas aeruginosa and Bacillus paramycoides within a sodium alginate (SA) matrix facilitated superior mass transfer of lipid globules (oleosomes), while ensuring high cell retention and protection against a harsh environment in the degradation system. Applying the PVA-SA composite yielded excellent results in successive degradation runs, i.e., up to 4 cycles. Analytical results from gas chromatography-mass spectrometry (GC-MS) justified the activation of the fatty aldehyde decarboxylase (FadD), fatty acyl-CoA reductase (Acr), and fatty aldehyde decarbonylase (CER1) enzymatic cascade. This activity resulted in the successful transformation of acyl groups into value-added hydrocarbons, specifically hexadecane and pentane. Detecting an m/z 115 peak (sodiated glycerol) served as a distinctive molecular marker for glycerol production. This SBW bacterial degradation achieved significant reductions in fat-oil-grease (FOG) = 95.18 f 3.81 %, chemical oxygen demand = 89.96 f 2.70 %, and total suspended solids = 86.15 f 3.45 %. The current results demonstrate the significance of coupling industrial wastewater remediation with the sustainable synthesis of bio-based chemicals (e.g., hexadecane, dodecane, and pentane). Additional studies should validate the Pseudomonas aeruginosa and Bacillus paramycoides performance for FOG degradation under industrial-scale applications, further encouraging the private sectors to invest in the simultaneous remediation of high-strength lipid-rich effluents and the recovery of high-value chemical precursors.

Aspergillus versicolor is a microorganism that widely contaminates cereals. It secretes a highly toxic substance, sterigmatocystin, and poses a great threat to the economy and human health. Biological antagonism by Bacillus subtilis HJ4 has been achieved in many ways, but the types of active antifungal substances it secretes during the process of biocontrol have not been completely analysed. Here, we analysed the metabolic pathways and products of B. subtilis HJ4 using multiomics technology and investigated the potential antifungal mechanism. The results showed that B. subtilis HJ4 exerted its biocontrol effect by secreting secondary metabolites with antifungal activity, including 4-phenylbutyric acid, isovaleric acid, and volatile organic compounds. In vitro experimental analysis showed that the antifungal compounds caused the leakage of intracellular nucleic acids and proteins by destroying the integrity of A. versicolor cells and spores. Furthermore, they inhibited the intracellular enzymatic activity of A. versicolor, caused metabolic disorders and oxidative damage to A. versicolor, and induced the apoptosis of A. versicolor cells, thus exerting antifungal effects. In summary, we explored new antifungal substances produced by B. subtilis HJ4 and the biological processes they affect. Moreover, we analysed the antifungal activities and potential applications of various compounds, providing a solid theoretical basis for the biological applications of B. subtilis HJ4.

Patulin (PAT) is a prevalent mycotoxin and environmental contaminant that poses serious threats to food safety, ecosystem stability, and human health. While previous studies have shown that intracellular enzymes from Geotrichum candidum XG1 can detoxify PAT, the specific enzyme responsible remained unidentified. In this study, proteomic analysis of Geotrichum candidum XG1 under PAT exposure revealed significant upregulation of redox-related proteins, implicating them in PAT detoxification. Molecular docking identified Geotrichum candidum XG1 aldo-keto reductase (GCAKR,initially annotated as YJR096W) as the top candidate enzyme, exhibiting the highest predicted binding affinity for PAT. Molecular dynamics simulations demonstrated the stability of the GCAKR-PAT complex, with a total binding energy of -16.57 kcal/mol. Key molecular components contributing to this stability included the ligand B:MOL-301, as well as protein residues TYR209 and PHE25. GCAKR was subsequently heterologously expressed in Escherichia coli, purified, and biochemically characterized. GCAKR exhibited optimal degradation activity at pH 8.0 and 30 °C, required NADPH as a coenzyme (≥1.0 mM), and achieved 100% degradation of 10 μg/mL PAT within 12 h at a concentration of 100 μg/mL. Its degradation efficiency was affected by enzyme/substrate concentration, with substrate inhibition observed at PAT concentrations ≥ 20 μg/mL. Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) analysis confirmed the conversion of PAT to E-ascladiol, a metabolite with low toxicity and non-carcinogenic properties. This study identifies GCAKR as the primary PAT-degrading enzyme in Geotrichum candidum XG1 and offers a mechanistic foundation for environmentally friendly mycotoxin detoxification strategies in biological systems.

Aflatoxin contamination caused by Aspergillus flavus in maize poses a major threat to food safety and public health. This study screened lactic acid bacteria (LAB) for anti-A. flavus activity and comprehensively evaluated their biocontrol efficacy, mechanisms, and application safety. Lacticaseibacillus paracasei F50 was isolated as a promising candidate, which exhibited pronounced condition-dependent and matrix-specific biocontrol performance: it strongly inhibited A. flavus in maize but showed minimal effects in peanuts. In maize, low LAB inoculation levels (1 × 106 cells/mL) reduced fungal biomass but increased aflatoxin accumulation, whereas high inoculation levels (1 × 109 cells/mL) suppressed both fungal growth and toxin production, achieving up to 90% inhibition of both fungal biomass and aflatoxin production. Mechanistic analysis revealed that L. paracasei F50 impeded spore germination and modulated the expression of key aflatoxin biosynthetic genes (aflR, aflS, aflD) and global regulators (veA, laeA), thereby interfering with the aflatoxin synthesis pathway. Genomic assessment and safety evaluation further confirmed its suitability as a biocontrol agent, and short-term application did not adversely affect maize quality (color, hardness, soluble solid content). Overall, L. paracasei F50 represents a viable microbial strategy for mitigating A. flavus contamination in maize, although long-term storage stability and large-scale field validation remain necessary for commercial application.

Patulin (PAT), a toxic mycotoxin from Penicillium species, contaminates fruits and fruit-based products, posing serious ecological and health risks. While enzymatic degradation offers an eco-friendly remediation approach, efficient degrading enzymes remain scarce. This study identified and characterized GCV2, a key PAT-degrading enzyme from Wickerhamomyces anomalus XL1, using transcriptomics, molecular docking, and molecular dynamics simulations. Transcriptomic analysis revealed oxidative stress response, detoxification, and glutathione metabolism as core PAT degradation pathways. Among 14 candidates, GCV2 showed the strongest PAT affinity (-6.31 kcal/mol). Molecular dynamics simulations confirmed GCV2-PAT complex stability, with electrostatic interactions driving binding and residues GLY607 and HIS738 providing stabilization. GCV2 was heterologously expressed and purified. Optimal conditions were 4.0 mM nicotinamide adenine dinucleotide phosphate (NADPH), pH 7.0-8.0, 30 °C, and 200 μg/mL enzyme, achieving complete degradation of 1-10 μg/mL PAT. Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) confirmed NADPH-dependent reduction of PAT to low-toxicity E-ascladiol and Z-ascladiol without toxic by-products. Notably, GCV2 efficiently degraded PAT in pear juice, achieving 85.72% degradation within 96 h. Physicochemical analysis, electronic nose, and gas chromatography-mass spectrometry (GC-MS) revealed that GCV2 preserved the inherent quality and flavor profile of the juice while significantly increasing total phenolics and vitamin C contents. Moreover, it enhanced key aroma components, including 4-Methyl-1-pentanol and nonanal, thereby enriching the fresh fruity and citrus-like aroma characteristics. This study identified GCV2 as an efficient PAT-degrading enzyme that preserves food quality, elevates nutrition and flavor, boasting great application potential in PAT detoxification of fruit juices and other fruit-based food products.

Traditional Egyptian cheese has a long history and holds an important place in the local culinary culture. However, systematic studies on the flavor compounds, microbial community structure, and their interrelationships in traditional Egyptian cheese remain limited. To ensure quality stability, this study systematically analyzed the characteristic flavor components and microbial community compositions of Mish and Karish cheeses by determining their physicochemical properties, organic acid content, and free amino acid content using headspace solid-phase microextraction gas chromatography-mass spectrometry and high-throughput sequencing techniques. The results showed that both were soft cheeses with high protein and low fat. Eight organic acids were detected, with lactic acid (70.54%) being the predominant component. Sixteen free amino acids were identified, of which essential amino acids accounted for 60.51%. Volatile compound analysis identified 86 volatile components, with 29 recognized as key aroma compounds (odor activity value ≥1). Among these, 2-nonanol, ethyl octanoate, ethyl decanoate, and ethyl hexanoate were common aroma compounds across all samples and formed the core aroma profile of traditional Egyptian cheeses. High-throughput sequencing revealed that the bacterial communities mainly consisted of Lactobacillus and Streptococcus, while the fungal communities were dominated by Kodamaea, Dipodascaceae, Torulaspora, and Metschnikowia, of which Torulaspora exhibited higher abundance in Karish cheese. Spearman correlation analysis indicated that Weissella and Kodamaea were closely associated with the development of cheese flavor, showing positive correlations with various amino acids, organic acids, and volatile compounds, and negative correlations with 2-dodecanol, 2-heptanol, isoamyl acetate, and ethyl acetate. Additionally, Streptococcus, Lactococcus, Candida, and Kluyveromyces also contributed significantly to cheese flavor development by participating in the synthesis of tartaric acid and certain ethyl esters. These findings provide novel insights into the microbial-driven flavor formation mechanism and a theoretical basis for screening industrial starter cultures and flavor regulation in traditional Egyptian cheese.

The increasing consumer demand for natural functional foods has prompted the development of fortified dairy products with improved nutritional and health benefits. This study designed and evaluated yoghurts fortified with cold-pressed moringa seed oil (MSO) and black seed oil (BSO) at 1.5 g /L, as well as their water-based nanoemulsions at 3 g Nanoemulsion powder/L, compared to a plain control. Phenolic profiles characterized by HPLC-MS revealed high levels of ellagic acid (23.5 mg/100 g), rutin (18.2 mg/100 g), and chlorogenic acid (21.4 mg/100 g) in MSO, and chlorogenic acid (19.8 mg/100 g), apigenin (16.7 mg/100 g), and naringenin (14.3 mg/100 g) in BSO. Nanoemulsions with 5% oil showed droplet sizes of 69.1 nm (MSO) and 38.1 nm (BSO) and zeta potentials above − 30 mV, confirming good colloidal stability over 7 days. Cytotoxicity assays indicated a safe dose up to 80 µg/mL. Yoghurt fortified with nanoemulsions exhibited a significant increase in total solids (+ 12%), unsaturated fatty acids (notably omega-3 increased by 25%), and antioxidant capacity (DPPH radical scavenging improved by 30%) while reducing acidity and syneresis relative to the control. Lactic acid bacteria viability remained unaffected. Sensory evaluation showed improved color, texture, and overall acceptance for yoghurts with 1.5 g nanoemulsion addition per liter. These findings demonstrate that incorporation of MSO and BSO nanoemulsions at this level effectively enhances yoghurt’s nutritional and functional properties without compromising microbial or sensory quality.

Abstract Up until now, most of the Ras cheese has been produced by small dairy sectors in Egypt using raw milk. This research aimed to determine the pathogens, and undesirable microorganisms in traditional Egyptian Ras cheese using classical methods and real-time PCR. Forty samples of Ras cheese were analysed for chemical and microbiological properties. The results revealed a wider range in the chemical composition of the collected samples. Most of the Ras cheese samples were within the Egyptian Standard for chemical compositions. There were only two samples of Ras cheese that had more moisture than the Egyptian Standard recommended.. The results of the microbiological analysis of the Ras cheese reveal that the coliform count ranged between 1 and 5.20 log CFU/g, with an average of 1.70 log CFU/g. Anaerobic spore-forming bacteria (Clostridium perfringens) were found in 29 out of 40 Ras cheese samples. All Egyptian cheese samples contained more yeast and mold than the Egyptian Standard recommends. All cheese samples were free of Listeria monocytogenes and Salmonella spp. The real-time PCR showed that 5% of samples were positive for methicillin-resistant Staphylococcus aureus, and all the analysed samples were free of E. coli O157:H7. According to this study's findings, most Ras cheese samples contained higher levels of Staphylococcus aureus, molds, and yeasts than what Egyptian standards recommended. The results of this study should be used to develop specific procedures for risk management along the milk production chain.

Correction for 'Screening of Lactococcus lactis strains with hypoglycemic effects and evaluation of their protective mechanisms against glucose-induced type 2 diabetes in zebrafish' by Huiying Li et al., Food Funct., 2025, 16, 6186-6202, https://doi.org/10.1039/D5FO01005E.

Hyperuricemia (HUA) is a major challenge in the field of public health, and long-term use of traditional drugs can easily lead to addiction and some side effects such as the hypersensitivity syndrome and liver and kidney damage. While several studies have proved that some probiotics are able to reduce serum uric acid (UA) levels, in this study, we found that Pediococcus acidilactici SWU-HX39 (HX39) has superior anti-HUA ability in vitro, and the therapeutic potential of HX39 in a HUA model of a high purine diet was also evaluated in vivo, focusing on its regulatory effects on liver xanthine oxidase (XOD) activity and serum proinflammatory cytokine and lipopolysaccharide (LPS) content. The results of the HUA mouse model showed a significant reduction of 49.8% in serum UA and 46.76% XOD activity by HX39 compared with the control group. Moreover, HX39 regulated the composition of the intestinal flora in mice, with significant decreases of the Bacteroides abundances and Parabacteroides, while Eubacterium_brachy_group and norank_f_Ruminococcaceae abundances increased. Further analysis showed that it helped alleviate inflammation and improve kidney metabolism. Significantly reduced kidney damage was observed in mice with HUA. These results may explain the potential mechanism of functional probiotics in alleviating HUA symptoms. Also, probiotic HX39 has promising applications in the treatment of HUA complications.

The contamination of spices with aflatoxin (AF) raises concerns regarding its health risks. In this study, 51 samples of dried red chili peppers (including 10 varieties collected from three regions) were used to investigate the AF contamination of dried red chili peppers sold in China, and the toxigenic characteristics of Aspergillus flavus strains isolated from chili peppers. AFs were identified using immunoaffinity column purification, derivatization, and high-performance liquid chromatography with fluorescence detection. The AF content in various types of dried red chili peppers sold in the Chinese market was generally low (≤7.19 µg/kg), below the limit of the relevant standard (10 µg/kg). The frequency of suspected A. flavus strains isolated from chili peppers was high (24/51), with a high toxigenic strain ratio of 29.17%. Further investigation revealed that 25-30 °C was the optimal temperature for toxin production by these toxigenic strains. This indicated that dried chilled peppers have a significant risk of AF contamination. Therefore, the findings of this study suggest that long-term storage of dried chili peppers should be conducted at temperatures below 25 °C or above 37 °C to mitigate the problem of AF contamination exceeding the levels allowed by the standards, providing new insights for the prevention and control of AF contamination of dried red chili peppers.

Aflatoxin contamination poses a significant food safety risk, particularly during the storage of dried chili peppers. This study evaluated the efficacy of formic acid treatment, ultraviolet (UV) treatment, and combined UV-formic acid treatment in both preventing and controlling Aspergillus flavus in dried red chili powder. Efficacy was assessed by measuring the growth diameter of A. flavus colonies on un-colonized and already colonized dried red chili powder. The optimal treatment conditions for the UV-formic acid combination were determined through single-factor experiments, orthogonal experiments, and quality assessment. Finally, the effects of the UV-formic acid combination on the cell membrane, antioxidant system, and energy metabolism of A. flavus were investigated. The results revealed that fumigation of un-colonized dried red chili powder with 5% formic acid for 24 h inhibited A. flavus growth by 93.29% and toxin synthesis by 99.41%. In contrast, treatment of already colonized chili powder with 10% formic acid inhibited A. flavus colony growth by 50%. Through a three-factor, three-level orthogonal experiment followed by quality testing, the optimal conditions were determined to be 8% formic acid concentration, a UV irradiation distance of 15 cm, and a treatment time of 75 min. This optimized combined treatment reduced the required fumigation time from 24 h to 1.25 h. This technique achieved complete suppression of aflatoxin B1 synthesis on un-colonized dried red chili powder. On already colonized chili powder, the mycelial growth inhibition rate was 48.05 ± 6.68%, and aflatoxin B1 synthesis was inhibited by 91.32 ± 3.15%. Quality assessment revealed that the UV-formic acid co-treatment parameters did not significantly affect key quality indicators including color, capsaicin content, total phenolic content (p > 0.05). Furthermore, UV-formic acid treatment disrupt the cell membrane structure of A. flavus, impairs its antioxidant and energy metabolism systems, and induces mitochondrial dysfunction. The study confirmed the synergistic antifungal effect of formic acid and UV, providing a potential industrialized solution for enhancing the safety and storage stability of dried chili products.

Background/Objectives: As probiotics gain prominence in the prevention and treatment of intestinal diseases, their protective effects against pathogens and influence on host health have drawn significant attention. This study investigates the genomic characteristics and functional potential of Pediococcus acidilactici XJ-24 (XJ-24) in the prevention of Listeria monocytogenes (LM) infection in mice. Methods/Results: Whole-genome analysis confirmed the safety and probiotic properties of XJ-24, including acid and bile salt tolerance, antimicrobial activity, and safety. In vivo, C57BL/6 mice challenges indicated that XJ-24 significantly reduced LM colonization, suppressed pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, IFN-γ), alleviated colon and spleen tissue damage, and maintained intestinal barrier integrity by upregulating tight junction proteins (Occludin, Claudin-1, ZO-1). Moreover, XJ-24 modulated gut microbiota composition by increasing beneficial taxa while reducing harmful bacteria. Correlation analysis highlighted a positive association between Lachnospiraceae and tight junction proteins. Conclusions: These findings demonstrate the potential of XJ-24 as a functional probiotic for preventing LM infection and provide a basis for further clinical exploration.

Patulin (PAT) is a neurotoxic fungal metabolite that commonly contaminates fruits and their derivatives, causing substantial economic loss and serious harm to human health. In this study, we screened a PAT-degrading yeast strain, Geotrichum candidum XG1, among 12 probiotic strains. It efficiently degraded 96.80 % of PAT (5 mu g/mL) within 48 h in potato dextrose broth and completely degraded PAT in fresh apple juice. G. candidum XG1 degraded PAT mainly by secreting intracellular enzymes, which were induced by PAT, and the degradation rate increased with increasing induction time. Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry identified ascladiol as a PAT degradation product that disappeared within 96 h. Notably, both G. candidum XG1 and its intracellular enzymes affected the quality of apple juice, and G. candidum XG1 enhanced the flavor of apple juice. Overall, these results highlight the potential of G. candidum XG1 to remove PAT from apple juice.