Search Results (1,109)

The growing global protein demand and environmental concerns from conventional animal agriculture have driven the exploration of sustainable alternative protein sources. Single-cell proteins (SCPs) from microbial fermentation offer a promising solution. This study comprehensively evaluated the nutritional value and safety profile of SCP produced from Ralstonia eutropha H16 through integrated in vitro and in vivo assessments. Nutritional analyses revealed a high crude protein content of 71.87 ± 5.05 g/100 g dry weight, with total amino acids of 53.67 ± 1.05 g/100 g. The essential amino acid content was 24.38 ± 0.51 g/100 g, accounting for 45% of the total amino acids. An essential amino acid index (EAAI) of 1.46 ± 0.04 and an amino acid score (AAS) of 0.83 ± 0.06 confirmed its classification as a high-quality protein source according to FAO/WHO standards. In vivo rat feeding trials demonstrated an adjusted protein efficiency ratio (PER) of 1.81, exceeding common plant proteins such as wheat (0.8–1.1). True digestibility (TD) reached 85.73%, with a biological value (BV) of 49.37%, net protein utilization (NPU) of 42.33%, and protein digestibility-corrected amino acid score (PDCAAS) of 0.71. Comprehensive safety assessments included chemical contaminant screening, acute oral toxicity studies in rats and mice, in vitro chromosome aberration tests, and erythrocyte micronucleus tests. Heavy metals and aflatoxin B₁ levels were below regulatory limits. Acute oral toxicity studies established LD₅₀ values exceeding 10,000 mg/kg body weight in both rodent species, classifying this protein source as practically non-toxic. The 28-day sub-acute toxicity study showed no significant adverse effects at low doses (6.25% protein replacement). Both genotoxicity assays (mammalian cell chromosome aberration assay and mammalian erythrocyte micronucleus test) returned negative results. These findings establish R. eutropha H16-derived SCP as a safe, nutritious, and sustainable protein source with considerable potential for feed and food applications, contributing to global food security and environmental sustainability. Full article

Aflatoxin B1 (AFB1) induces hepatocellular damage through its metabolite aflatoxin B1-8,9-epoxide (AFBO), which is produced in endoplasmic reticulum (ER) via cytochrome P450 (CYP450) enzymes. To investigate the effect of ferulic acid (FA) on AFB1-induced broiler liver damage, one-day-old Arbor Acres broilers were exposed to AFB1 (4 mg/kg) and treated with different doses of FA (60 mg/kg, 120 mg/kg, and 240 mg/kg) continuously for 28 days. The production performance, biochemical indicators, morphological changes, CYP450 enzymes’ expression in ER, interactions between small molecules and CYP450 enzymes, and CYP450 enzymes’ protein secondary structure were investigated. The results showed the following: (I) FA promoted broiler growth and reduced AFBO production. (II) AFB1-induced changes in serological indicators (AST, ALT, ALP, γ-GT, TBA, TG) and biochemical parameters (GST, SOD, MDA, ROS), which were reversed by FA. (III) AFB1-induced liver morphological changes and apoptosis were obviously alleviated by FA. (IV) AFB1-induced up-regulation of CYP1A5, CYP2A6, CYP2W1, and CYP3A4 in ER were reduced by FA. (V) The binding affinity of FA to CYP1A5 is lower than that of AFB1 to CYP1A5, and the binding affinity of FA to CYP2W1 is similar to that of AFB1 to CYP2W1. (VI) The contents of α helix, β sheet, β turn, and random coil in chicken CYP1A5 were 59.6%, 7.8%, 13.6%, and 19.0% respectively, and those in chicken CYP2W1 were 32.2%, 17.1%, 16.8%, and 33.9% respectively. In conclusion, FA can promote broiler growth and alleviate AFB1-induced hepatotoxicity via inhibiting and conjugating CYP450 enzymes, thus reducing AFBO formation and oxidative damage. Full article

Alternative protein sources are increasingly considered in poultry nutrition, and insect meals may represent a promising strategy for modifying the compositional profile of maize silage in the context of waterfowl feeding. This study evaluated maize silage supplemented with Hermetia illucens (BAHI) or Tenebrio molitor (BATM) meal as alternatives to urea (BAUR) as a nitrogen source, including variants with lauric acid (BALA) and with the lactic acid bacteria (LAB) inoculant alone (BA), focusing on nutritional composition, microbiological quality, and selected mycotoxins (MT). Maize forage treated with LAB was ensiled for 8 weeks in five variants, with three independent silage replicates per treatment, and the silages were analyzed for nutrients, energy value, amino acids, fatty acids, microbial and MT content. Insect meals increased crude protein compared with BA (64.94–67.32 vs. 49.73 g/kg dry matter, DM) and nearly doubled ether extract in BAHI (40.08 vs. 21.74 g/kg DM), while apparent metabolizable energy for poultry ranged from 8.55 to 9.87 MJ/kg DM (BAHI). Urea elevated crude protein content to 66.37 g/kg DM without improving the essential amino acid index. Microbial indicators did not differ among treatments. In the MT profile, lower deoxynivalenol concentrations were observed in BAUR, BAHI, and BATM, and lower fumonisin B1 concentrations were observed in BATM and BALA, while aflatoxins and ochratoxin A were not detected. Overall, the results indicate that low-level inclusion of insect meals in combination with LAB improved the compositional characteristics of maize silage, while no significant differences were detected in the monitored microbial counts and only selective changes were observed in the mycotoxin profile. Full article

Multiple mycotoxins in feed threaten animal health and food safety, demanding sustainable mitigation strategies. This study evaluated acid-modified mangosteen peel (AMP), an agricultural by-product, as a potential multi-mycotoxin adsorbent. Physicochemical characterization using scanning electron microscopy (SEM), Brunauer–Emmett–Teller (BET) surface area analysis, and Fourier transform infrared spectroscopy (FTIR) analyses demonstrated that acid modification increased surface area (1.9 to 9.03 m²/g), pore volume (0.005 to 0.027 cm³/g), and surface negativity, indicating enhanced adsorption properties. In vitro binding experiments assessed adsorption of aflatoxin B₁ (AFB₁), zearalenone (ZEA), ochratoxin A (OTA), T-2 toxin, deoxynivalenol (DON) and fumonisin B₁ (FB₁) under different pH conditions. AMP exhibited high adsorption efficiencies for AFB₁, ZEA, OTA, and T-2 toxin, particularly at pH 3, whereas DON and FB₁ showed limited binding. Adsorption behavior was dose-dependent and best described by Langmuir and Freundlich isotherm models. Simulated gastrointestinal digestion indicated stable binding of AFB₁ and ZEA under gastric conditions, with partial release of some toxins at neutral pH. Cytotoxicity assessment in porcine intestinal epithelial cells (IPEC J2) showed no apparent cytotoxic effects at 0.25–1 mg/mL. Therefore, AMP demonstrated improved multi-mycotoxin adsorption compared to the untreated material and showed no apparent cytotoxic effects in vitro within the tested concentration range, indicating its potential as a promising feed additive candidate. Full article

Aflatoxin B1 (AFB1) is a prevalent and highly toxic mycotoxin in the food and feed chain and can directly injure the intestinal epithelium. Yet, its upstream determinants linking epithelial stress to cytotoxicity remain insufficiently defined. Here, we used porcine intestinal epithelial IPEC-J2 cells to characterize AFB1-induced cytotoxic and transcriptomic responses and to determine the role of the tight-junction scaffold, Cingulin-like 1 (CGNL1), a candidate gene identified through genome-scale CRISPR knockout library screening. The results showed that AFB1 exposure reduced cell viability in a dose-dependent manner and induced oxidative stress. RNA-seq profiling analysis revealed broad transcriptional remodeling, with activation of inflammatory pathways (including NF-κB and JAK–STAT signaling). Based on our constructed CGNL1-knockout IPEC-J2 cell line (CGNL1-KO IPEC-J2) using CRISPR/Cas9, it was found that CGNL1 deficiency markedly alleviated AFB1-induced cytotoxicity and oxidative stress. Comparative transcriptomics analysis showed that CGNL1 knockout attenuated AFB1-triggered aberrant expression of some CGNL1-dependent AFB1-responsive genes related to immune response under AFB1 challenge. Together, these findings identify CGNL1 as a potential modulator of epithelial susceptibility to AFB1 and support its involvement in the regulation of toxin-induced oxidative response. Full article

Mycotoxin contamination is an important threat to food and feed safety as well as human and animal health, with particular emphasis on oxidative stress, apoptosis, autophagy, inflammation, and dysbiosis. Mycotoxins represent major health threats because they disturb cellular homeostasis and induce oxidative damage. Nutritional factors, such as dietary antioxidants and bioactive chemicals, can influence the body’s reaction to mycotoxin exposure, either reducing or increasing its effects. This study discusses how mycotoxins (aflatoxin B1, deoxynivalenol, and ochratoxin A) induce oxidative stress by producing reactive oxygen species (ROS)-mediated DNA damage, which induces cellular damage and activates apoptosis, an intended cell death process that is critical for tissue integrity. Furthermore, mycotoxins alter autophagy, a cellular degradation process that can be beneficial or destructive depending on the situation, affecting cell survival. The inflammatory response is particularly important because mycotoxin-induced oxidative stress and cell damage activate inflammatory pathways, which contribute to tissue injury and disease progression. Nutritional factors high in antioxidants, anti-inflammatory substances (Lycopene, Curcumin, Thyme oil, Gum Arabic, and Ginger), probiotics, and prebiotics show potential in mitigating these negative consequences by reducing oxidative stress and inflammation. Advances in molecular biology and omics technologies (transcriptomics, proteomics, metabolomics, and single-cell sequencing) can lead to better knowledge of the underlying pathways, allowing for more tailored nutritional recommendations and medicinal interventions. Finally, combining dietary modulation with mycotoxin risk management is a viable path for protecting health and increasing resilience to mycotoxin-related toxicities in animals. Full article

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

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

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 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