Search Results (1,799)

Aflatoxin B1 (AFB1) and patulin (PAT) are prevalent foodborne mycotoxins with hepatotoxic potential, but the hepatic effects of combined exposure remain largely unclear. This study investigated the hepatotoxic consequences of co-exposure to AFB1 and PAT using no-observed adverse effect levels (NOAELs) in C57BL/6 mice and low-cytotoxic concentrations in HepG2 cells selected by viability screening. Mice and cells were assigned to four groups: control, AFB1, PAT and AFB1 + PAT. Exposure to either toxin individually did not cause evident liver injury, whereas co-exposure significantly elevated alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, reduced liver index, and induced clear histopathological alterations. Co-exposure markedly aggravated oxidative stress, characterized by increased reactive oxygen species (ROS) and malondialdehyde (MDA) and decreased superoxide dismutase (SOD). In parallel, the levels of interleukin-6 (IL-6), interleukin-1 beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) were elevated, together with the early fibrosis-related markers alpha-smooth muscle actin (α-SMA) and vimentin. The apoptotic response was characterized by increased Bcl-2-associated X protein (Bax) and reduced B-cell lymphoma-2 (Bcl-2), together with cysteine-dependent aspartate-specific protease-3 (caspase-3) activation. These findings indicate that co-exposure to AFB1 and PAT elicits hepatotoxicity through amplified oxidative stress, inflammation, and caspase-dependent apoptosis, supporting the need to further consider mycotoxin co-exposure in toxicological evaluation. Full article

Aflatoxin B₁ (AFB₁) is a highly stable mycotoxin that can persist during conventional food processing and therefore poses a serious risk to food and feed safety. In this study, two enzymes (CotA and Prx) were heterologously expressed in Bacillus subtilis, purified by Ni–NTA affinity chromatography, and evaluated for their ability to degrade AFB₁. Both enzymes exhibited remarkable thermostability and distinct catalytic optima. CotA exhibited its highest activity at 80 °C with an AFB₁ removal of 38.4%, whereas Prx showed its highest activity at 90 °C with a removal of 82.6%. The optimal pH values were near neutral, with CotA performing best at pH 7.0 and Prx at pH 7.5, and both reactions approached maximal conversion within approximately 10 h. When the two enzymes were combined, a clear cooperative effect was observed. The mixed system achieved 91.0% AFB₁ removal at 80 °C after 10 h, with the best degradation activity occurring at a CotA to Prx ratio of 1:3. At 50 °C, neither enzyme alone caused appreciable AFB₁ degradation, but the addition of hydrogen peroxide markedly enhanced catalytic activity. Both enzymes also retained substantial activity after boiling and autoclaving. In a maize flour model, the mixed-enzyme system showed strong AFB₁ degradation capacity, and peroxide-assisted treatment further improved activity. These results establish a thermostable and peroxide-responsive enzymatic platform for AFB₁ degradation and support future development of enzyme-based detoxification strategies for food and feed applications. Product identification and toxicological validation will be needed to confirm the safety of the treated products. Full article

Aflatoxin B1 (AFB1) is a potent mycotoxin threatening food and feed safety. Here, we report the identification and characterization of a Bacillus safensis-derived multicopper oxidase (BsaMCO) capable of efficient AFB1 detoxification. Recombinant BsaMCO exhibited robust in vitro activity, achieving >78% degradation of AFB1 under 24 h incubation at 37 °C. Optimization experiments revealed that enzyme concentration, pH, temperature, metal ions, and electron acceptors significantly influenced degradation efficiency, defining an operational window suitable for practical applications. LC–MS profiling suggested the presence of transformation products tentatively consistent with oxidative demethylation to aflatoxin P1 (AFP1) and with the formation of AFG2a-like products through subsequent hydration- and oxidation-related transformations. Molecular docking and 100 ns all-atom molecular dynamics (MD) simulations demonstrated stable binding of AFB1 in the T1 copper pocket. Van der Waals and electrostatic interactions, together with a persistent hydrogen bond at Gly323, facilitated single-electron transfer through the intramolecular T2/T3 copper cluster. Principal component and Gibbs free energy analyses confirmed a low-energy, stable conformational ensemble. HepG2 cell assays indicated that BsaMCO-degraded products substantially reduced cytotoxicity and apoptosis compared with native AFB1. Simulated feed experiments further validated enzymatic AFB1 degradation, with approximately 53% reduction after 24 h. Collectively, these findings establish BsaMCO as a safe and effective biocatalyst for AFB1 detoxification, providing mechanistic, structural, and cellular evidence supporting its application in food and feed safety. Full article

Basil essential oil (BEO) has emerged as a promising natural alternative to antibiotic growth promoters in poultry production. BEO has shown antimicrobial, antifungal, anticoccidial, antioxidant, and insecticidal properties. BEO exhibits broad antimicrobial activity against Gram-positive and Gram-negative pathogens, and modulates gut microbiota by decreasing Escherichia coli and Staphylococcus spp. Anticoccidial effects include reduced oocyst shedding, improved intestinal morphology, and downregulation of pro-inflammatory cytokines. Antifungal activity reduces fungal load and inhibits Aspergillus spp., with implications for control spoilage and aflatoxin risk. BEO at a concentration of 40 ppm was effective in preventing E. tenella invasion, showing an average reduction in invasion by 36% in primary chicken epithelial cells. Antioxidant benefits include enhanced intestinal and systemic antioxidant status. Advanced nanoformulation technologies, particularly nano-encapsulation, have substantially overcome several limitations for BEO application in poultry. Further research is still required to assess the efficacy of nano-encapsulated BEO for enhancing overall poultry industry productivity. This review synthesizes current evidence on BEO integration in the poultry production sections, from nutrition and disease control to product preservation and farm hygiene, and evaluates technological solutions that address formulation barriers. Moreover, it discusses critical research gaps and proposes future directions for enhancing BEO applications in sustainable poultry production systems. Full article

Aspergillus flavus is a globally distributed filamentous fungus of major agricultural and medical importance, capable of producing carcinogenic aflatoxins and forming two specialized developmental structures, conidia and sclerotia. While the molecular framework governing conidiation has been well characterized in Aspergillus nidulans, the corresponding mechanisms in A. flavus remain somewhat unelucidated. In this study, we identified and functionally characterized AfldrnA, a gene encoding a basic helix–loop–helix (bHLH) transcription factor. Targeted deletion of AfldrnA resulted in an aconidial phenotype accompanied by a significant increase in sclerotia formation, whereas complementation with the intact AfldrnA gene restored conidiation and reduced sclerotia development. Phenotypic assays revealed that the ΔAfldrnA mutant exhibited normal vegetative growth, unchanged antifungal susceptibility, and unaffected aflatoxin B₁ production, indicating that AfldrnA primarily regulates developmental rather than metabolic differentiation. Additionally, observed differences between standard and dark incubation conditions suggest that AfldrnA may be involved in environmentally responsive regulation of fungal development. Overall, this study identifies AfldrnA as a pivotal transcriptional regulator essential for coordinating conidiation and sclerotia formation in A. flavus, providing new insights into the genetic and environmental regulation of fungal developmental programs. Full article

This study aimed to estimate dietary exposure to aflatoxins (AFs) and characterize its associated risks through the consumption of dried fruits and chocolates among the adult population of Yerevan, the capital city of Armenia. Asflatoxin B1 (AFB1) and total AFs were determined using HPLC in 10 composite samples of widely consumed dried fruits and chocolates, prepared by pooling 100 individual sub-samples into 5 dried fruits and 5 chocolate composites. Individual consumption data were obtained via food frequency questionnaires and were stratified by consumer groups and percentiles. Exposure scenarios (lower-, middle-, upper-bound and detected mean) were applied, and risk was assessed using the margin of exposure (MOE) approach with a BMDL₁₀ of 0.4 μg/kg bw/day. The study findings revealed that dried fruits had higher contamination levels (detected mean content of 10 μg/kg AFB1, 15 μg/kg total AFs) compared to chocolates (detected mean content of 0.5 μg/kg AFB1, and 0.9 μg/kg total AFs), resulting in lower MOE values despite smaller consumption quantities. Detectable AFs in dried fruits from open (street) markets exceeded the EU maximum limits, while Armenia currently lacks national regulatory limits for these products. MOEs were below 10,000 for most consumption groups, indicating a potential public health concern. This research emphasizes the urgent need for continuous monitoring and the establishment of harmonized national regulatory limits for AFs in dried fruits. Full article

Mycotoxin contamination remains a persistent threat to food safety in the Democratic Republic of the Congo (DRC) and neighboring countries, driven by conducive tropical agroecological conditions, inadequate post-harvest practices, and limited regulatory governance. This critical narrative review (2009–2024) synthesizes the occurrence data for major staple foods (maize, peanuts, cassava, sorghum, millet, and beans) and dairy products compiled from Google Scholar, ScienceDirect, MDPI and institutional sources. It examines the co-occurrence patterns, exposure pathways, and analytical and regulatory gaps. Warm, humid lowland environments favor Aspergillus and aflatoxins, whereas cooler, humid highland zones promote Fusarium, fumonisins, and deoxynivalenol. Across commodities, contamination intensifies along food value chains through inadequate drying, non-hermetic storage, insect damage, and prolonged handling, with processed products generally exhibiting the highest levels of mycotoxins. Regulated mycotoxins, including aflatoxins, fumonisins, trichothecenes, ochratoxins, and zearalenone, frequently exceed European Union (EU), East African Community (EAC), and Codex Alimentarius Commission (CAC) limits in staple foods. Their co-occurrence is widespread, including emerging mycotoxins such as beauvericin and enniatins, particularly in maize- and peanut-based products, raising concerns about potential additive or synergistic effects. Aflatoxin M1 in milk highlights plant–feed–animal–human transfer within a One Health framework. Despite increasing evidence, the available data remain fragmented and heterogeneous; rapid tests dominate, while few studies employ multi-mycotoxin LC-MS/MS methods. Cross-border trade between countries, such as Uganda, Tanzania, Zambia and Angola, facilitates the circulation of contaminated commodities in the absence of harmonized standards and risk-based controls. Priorities include harmonized regional surveillance, biomarker-based co-exposure assessment, cost-effectiveness evaluation of mitigation strategies, and regulatory alignment at borders. Coordinated, multisectoral action is essential to reduce chronic dietary exposure and improve food safety across the region. Full article

Aflatoxin B1 (AFB1) poses a serious threat to global food and feed safety. Laccase-based enzymatic degradation represents a promising green strategy for AFB1 removal; however, its industrial application is severely limited by the rapid thermal inactivation of wild-type enzymes under high-temperature processing conditions (>70 °C). Here, we engineered the thermal stability of a laccase from Bacillus amyloliquefaciens B10 through an integrated strategy combining computational structural biology with semi-rational design. By coupling molecular dynamics (MD) simulations with folding free-energy (ΔΔG) calculations, we identified key flexible regions associated with thermal instability and subsequently implemented iterative saturation mutagenesis. The best single mutant, R196C, retained more than 96% relative activity after heat treatment at 80 °C for 10 min. Further iterative mutational stacking progressively enhanced thermostability: the R90E/R196C double mutant showed 1.25-fold higher activity at 80 °C than R196C, and the R90E/R196C/H54F triple mutant showed a further 1.16-fold increase over the double mutant. The final quadruple mutant, R90E/R196C/H54F/R253I, achieved 86.9% AFB1 degradation at 80 °C after 24 h. High-temperature MD simulations (100 ns at 353.15 K) indicated that the enhanced thermostability was associated with reduced conformational flexibility, lower radius of gyration (Rg) and solvent-accessible surface area (SASA), and a coil-to-β-sheet transition that contributed to stabilization of the protein core. In addition, efficient secretory expression of the engineered enzyme was achieved in Pichia pastoris, reaching 3.0 U/mL, while the crude enzyme maintained more than 70% activity at 80 °C. Collectively, these results provide a practical basis for the rational engineering and scalable production of thermostable biocatalysts for AFB1 detoxification-related applications of AFB1 control, and offer broader insights into the targeted enhancement of thermal stability in industrial enzymes. Full article

The effects of broad-spectrum mycotoxin detoxifiers (BSMDs) on growth performance, liver histopathology, jejunal morphology, and oxidative stress were evaluated in broilers fed diets contaminated with multiple mycotoxins. A total of 800 one-day-old male Ross 308 broilers were randomly assigned to four treatments, with eight replicates of 25 birds each, and reared for 42 days. The treatments included a basal control diet (CON); a multi-mycotoxin-contaminated diet (MMT) containing aflatoxins (25 µg/kg), zearalenone (135 µg/kg), T2 toxin (85 µg/kg), fumonisin (1.90 mg/kg), and deoxynivalenol (0.70 mg/kg); and the MMT diet supplemented with either 1.0 kg/ton BSMD-1 or 1.5 kg/ton BSMD-2. MMT contamination did not affect growth performance, serum malondialdehyde, interleukin-6 levels, liver enzyme activities, or liver lesion scores. Nevertheless, interleukin-10 levels were lower in birds fed the MMT diet (p = 0.03). In birds fed MMT, there was a substantial decrease (p < 0.05) in the height of the jejunal villi, their surface area, and the ratio of their height to the depth of the crypt. While BSMD-supplemented groups displayed values similar to both CON and MMT, MMT birds had higher fatty liver scores than the control group. Overall, multi-mycotoxin contamination impaired gut morphology and immune balance. BSMD supplementation improved intestinal structure, enhanced immune response, and partially mitigated liver alterations. These findings indicate its potential as a dietary intervention to mitigate the detrimental effects of multi-mycotoxin contamination in broilers. Full article

Highly sensitive methods for trace-level aflatoxin determination are indispensable for cereal food safety and public health protection. This study developed a ZrO₂-assisted QuEChERS-UHPLC-MS/MS method for the simultaneous determination of AFB₁, AFB₂, AFG₁, and AFG₂ in maize, wheat, rice, and soybean. Systematic optimization identified acetonitrile as the optimal extraction solvent and 10 mg ZrO₂ in combination with PSA, C₁₈, and GCB as the optimal cleanup formulation, providing recoveries of 107.33–111.60%. Chromatographic baseline separation was achieved within 8.0 min using a moderate gradient program. The method exhibited excellent linearity (R² > 0.999) with LODs of 0.15–0.25 µg/kg and LOQs of 0.50–0.75 µg/kg. Negligible matrix effects (0.85–1.02) validated the efficacy of ZrO₂-assisted cleanup in eliminating co-extractive interferences in maize. Satisfactory accuracy (recoveries of 86.66–111.04%) and precision (RSDs < 14%) were obtained across all matrices. The method demonstrated consistent performance across diverse cereal and soybean matrices, fulfilling international regulatory requirements for routine aflatoxin monitoring in agricultural commodities. Full article

The use of agro-industrial waste to obtain biochar has emerged as an environmentally friendly, low-cost, effective, profitable, and sustainable strategy for the removal of aflatoxin B1 (AFB1), a highly toxic and carcinogenic mycotoxin of importance in poultry production systems because it can cause serious economic losses, affect hatchability, egg production, and the growth of birds, and can cause their death. In this sense, the objective of the present study was to obtain a sustainable and low-cost biochar derived from agro-industrial coconut shell waste (BCS) and evaluate its AFB1 adsorption capacity using a conventional method based on buffer solutions and an in vitro avian digestion model that simulates the conditions of the gastrointestinal tract of the broiler chicken. The results showed that the adsorption capacity of BCS on AFB1 (250 ng/mL) at both pH 5.0 and 1.2 was close to 100%, while at pH 6.8, the adsorption of AFB1 was 86.24%. However, in the in vitro avian digestibility model, the adsorption capacity of BSC on AFB1 was 32.96%, thus highlighting the importance of considering factors that can affect the adsorption capacity of materials before in vivo studies, as this can lead to overestimations of results and, therefore, ineffective treatments or unexpected results in animals. Full article

In the present study, an amperometric aflatoxin B1 sensor was constructed via modifying a nano-graphite carbon paste microelectrode with a cationic surfactant of cetyltrimethylammonium bromide (CTAB) and a perfluorosulfonic acid resin of Nafion through a simple and controllable electrochemical scanning method. The experiment results show that CTAB–Nafion composite film has a good catalytic effect on the electrochemical response of aflatoxin B1. The electrocatalytic mechanism was investigated with the aid of different analytical techniques, including square wave voltammetry, electrochemical impedance spectroscopy, chronocoulometry, energy-dispersive spectroscopy and scanning electron microscopy. Under the optimal conditions, the linear range of the sensor is from 0.1 nM to 100 nM, and its detection limit and sensitivity are 20 pM (S/N = 3) and (24.9 ± 1.51) μA/nM, respectively. The accurate and rapid detection of aflatoxin B1, which has strong carcinogenicity, is of great significance for food quality monitoring and the protection of human health. Therefore, finally, the sensor was used to detect the concentration of aflatoxin B1 in milk and soy sauce samples, and the favorable recovery results indicated its good application prospects. Full article

This study provides a combined profile of fungal isolates from fresh and dried chili peppers in markets in Guangzhou. Multilocus sequence analysis revealed a wide variety of species, seven of which were reported for the first time from chili pepper (F. annulatum, F. compactum, F. pernambucanum, F. ramsdenii, and F. tardichlamydosporum, P. citrinum and P. steckii). In this research work, quantitative determination using targeted LC–MS/MS of dried chili peppers showed a significantly higher frequency of contamination and higher toxin concentrations than fresh samples. The predominant mycotoxins in dried peppers were DON and FB₁, which were present in all the samples at mean levels of 0.56 µg/g and 0.067 µg/g, respectively. AFB₁ and OTA were present in all dried samples but were detected only occasionally in fresh peppers. ZEN and CIT were detected at lower concentrations, but more prevalent among dried products (63.6% and 81.8% of all samples, respectively). The aflatoxin B₁ (AFB₁) level of 180 µg/kg in dried chili samples was 36 times above the EU maximum limit (5 µg/kg), and the OTA level reached 54 µg/kg, exceeding the EU limit by a factor of 2.7 (20 µg/kg). Statistical analysis also showed that all six mycotoxins were statistically higher in dried pepper than in fresh pepper. In vitro evaluation demonstrated that certain Fusarium isolates synthesized FB₁. At the same time, Penicillium species, including P. citrinum and P. steckii, consistently produced citrinin, confirming the strong influence of growth substrate on toxin biosynthesis. The frequent occurrence and elevated levels of regulated mycotoxins highlight significant public health concerns and underscore the need for improved postharvest handling and drying practices. These findings provide critical baseline data linking fungal diversity with toxin production dynamics, developing essential guidance for targeted mitigation strategies. Full article

Human organoids and organ-on-chip/microphysiological systems (OoC/MPS) are increasingly used as new-approach methodologies for biotoxin assessment. They retain human-relevant tissue organization and enable interpretable analysis of exposure geometry, barrier transport, perfusion, and (when needed) multi-organ coupling. In this review, we synthesize primary evidence across major toxin classes, including bacterial enterotoxins (e.g., cholera toxin, heat-stable enterotoxins, Shiga toxins), mycotoxins (e.g., aflatoxin B1, ochratoxin A, deoxynivalenol), and algal/cyanobacterial toxins (e.g., saxitoxin, domoic acid, microcystins, biliatresone). We emphasize studies that clearly define toxin identity and exposure context and that demonstrate mechanism-critical model competencies under assay conditions. We highlight decision-informative functional endpoints that align with the dominant pathophysiology. These include cystic fibrosis transmembrane conductance regulator (CFTR)-dependent secretion in human enteroids/colonoids, transporter-linked proximal tubular injury in kidney MPS, gut–kidney axis injury from Shiga toxin-producing E. coli in microfluidic systems, and multi-electrode array (MEA) network readouts in human 3D neural tissues. We then summarize best practices that improve cross-study comparability. These include reporting delivered versus nominal exposure, assessing recovery/mass balance and device/material interactions, applying proportional biological qualification (polarity, transporter/enzymatic competence, functional stability), defining a minimal comparable endpoint core, and preserving QIVIVE readiness in reporting. Finally, we outline near-term priorities for the field, including chronic low-dose and mixture designs, harmonized reference panels and acceptance criteria, and fit-for-purpose escalation to coupled OoC/MPS only when perfusion or organ–organ coupling is expected to change the interpretation. Full article

The objective of this study was to evaluate the preharvest application of γ-aminobutyric acid (GABA), melatonin (MT), and oxalic acid (OA), at different concentrations and application frequencies, on the physicochemical and microbiological quality of dried figs (cv. Calabacita) at commercial harvest and after 3 and 6 months of refrigerated storage. A further aim was to determine their impact on fungal populations and mycotoxin production. The results showed that untreated dried figs had a higher frequency of Aspergillus welwitschiae, A. tubingensis, and Aspergillus section Flavi, whereas elicitor-treated figs exhibited a lower incidence of toxigenic fungi. A. welwitschiae was the main ochratoxin A (OTA)-associated species detected, although the proportion of OTA-positive figs was lower in elicitor-treated samples than in the control. Aflatoxins (AFs) were detected only sporadically in 2 mM OA treatments, consistent with the limited activity of A. flavus at low storage temperatures. Conversely, Penicillium spp. were widespread but were associated with citrinin (CIT) production only under 2 mM OA treatments. Among the Alternaria toxins, alternariol (AOH) was detected solely in dried figs treated with 1 mM OA. Notably, all investigated mycotoxins were below the limit of detection (<LOD) in dried figs treated with 0.5 mM MT. Moderate elicitor concentrations (e.g., 0.5 mM MT and 50 mM GABA) and multiple preharvest applications generally provided the best balance between fungal suppression and fruit quality, significantly reducing Aspergillus spp. occurrence without promoting the growth of undesirable species. Overall, elicitor treatments decreased the incidence of toxigenic fungi, most likely through direct antifungal effects in senescent dried fruit rather than by inducing host defences. The combined use of preharvest elicitors with appropriate drying and storage conditions is a promising strategy to control fungal contamination and mycotoxin accumulation in dried figs while maintaining quality from preharvest storage. Further research is needed to optimise elicitor concentrations and application timing. Full article