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Aflatoxin B1 (AFB1) is a highly toxic carcinogen severely harmful to humans and animals. This study fabricated SDB-6-K-9@TiO2 composites via the hydrothermal synthesis method to reduce AFB1. The structural characterization results of the photocatalytic composites showed that TiO2 was successfully loaded onto SDB-6-K-9. The different photocatalytic degradation conditions, photocatalyst kinetics, recycling performance, and photocatalytic degradation mechanism were investigated. Photocatalysis with 6 mg of 4%SDB-6-K-9@TiO2 in a 100 μg/mL AFB1 solution presented a reduction of over 95%, exhibiting excellent performance, high stability, and reusability even after five cycles of photocatalytic experiments. Active species trapping experiments confirmed that holes (h+) played the most critical role. After structural analysis and identification of the photocatalytic degradation products, the photodegradation path and photocatalytic oxidation mechanism of 4%SDB-6-K-9@TiO2 were postulated. The results show a new way to improve TiO2’s photocatalytic performance, providing a certain theoretical basis for the effective AFB1 reduction.

Aflatoxin B1 (AFB1) produced by some Aspergillus species belongs to the most dangerous contaminants of animal feeds. Development of safe and cost efficient decontamination methods saving feed quality and nutritional value are of paramount importance. The use of recombinant AFB1-detoxifying microbial enzymes represents a promising biotechnological approach meeting the aforementioned requirements. In this study, three AFB1-degrading oxidases (AFOs) from edible basidiomycetes Cantharellus cibarius, Lentinula edodes and Pleurotus eryngii as well as AFO from Armillaria tabescens were expressed in E. coli Rosetta (DE3) and purified by immobilized metal-chelate chromatography. The stabilizing effect of the addition of glycerol and β-mercaptoethanol during protein extraction is shown. The catalytic constants of the recombinant AFOs (rAFOs) and other characteristics, which might be important for their practical application (and optimal temperature and pH, thermolability, regulation of the activity by metal ions and chelating agents, storage stability) were investigated. Among the obtained enzymes, rAFO from P. eryngii (Pe-AFO), which was characterized by the highest specific activity, thermostability and pH stability (especially at acidic pH range), the lowest Km, and relative resistance to the inhibition by phytate, showed the best AFB1-degrading efficacy. However, Pe-AFO and all other rAFOs significantly decreased the target activity during heating above 45 °C, storage frozen or lyophilization.

Aflatoxins belong to a class of mycotoxins, among which aflatoxin B1 (AFB1) has detrimental effects on the health of both animals and humans. It is associated with long-term exposure-induced carcinogenicity, hepatotoxicity, renal toxicity, neurotoxicity, and immunosuppressive properties, resulting in a variety of diseases. The intestine is the first barrier for human exposure to AFB1, but limited investigations have been conducted to clarify the underlying mechanisms of intestinal cytotoxicity. The mechanism of AFB1-induced cytotoxicity was investigated in this study using an integrated approach combining transcriptome, proteome, and metabolome analysis along with molecular dynamics simulation. After exposing SW480 cells to 50 μM AFB1 for 72 h, the transcriptome, proteome, and metabolome exhibited significant enrichment in pathways associated with oxidative stress, fatty acid and lipid metabolism, and glutathione metabolism. The experimental results demonstrated that AFB1 significantly reduces SW480 cells viability, and induces oxidative stress, calcium overload, mitochondrial damage, and lipid metabolism disorders.

This study aimed to assess the effectiveness of aflatoxin B1 (AFB1) and Thunbergia laurifolia extract (TLE) in the diets of Cherry Valley ducklings. Our investigation covered growth indicators, blood biochemical indices, meat quality, intestinal morphology, immune response, and CP450 enzyme-related gene expression. We conducted the study with 180 seven-day-old Cherry Valley ducks, randomly divided into five dietary treatments. These treatments included a basal diet without AFB1 (T1 group), TLE, or a commercial binder; the basal diet containing 0.1 mg AFB1/kg (T2 group), 0.1 mg AFB1/kg and 100 mg TLE/kg (T3 group), 0.1 mg AFB1/kg and 200 mg TLE/kg (T4 group), and 0.1 mg AFB1/kg and 0.5 g/kg of a commercial binder (T5 group), respectively. Ducklings fed with the T2 diet exhibited lower final body weight (BW), average body weight gain (ADG), and poor feed conversion ratio (FCR) during the 42-day trials. However, all ducklings in the T3, T4, and T5 groups showed significant improvements in final BW, ADG, and FCR compared to the T2 group. Increased alanine transaminase (ALT) concentration and increased expression of CYP1A1 and CYP1A2 indicated hepatotoxicity in ducklings fed the T2 diet. In contrast, ducklings fed T3, T4, and T5 diets all showed a decrease in the expression of CYP1A1 and CYP1A2, but only the T4 treatment group showed improvement in ALT concentration. AFB1 toxicity considerably raised the crypt depth (CD) in both the duodenum and jejunum of the T2 group, while the administration of 200 mg TLE/kg (T4) or a commercial binder (T5) effectively reduced this toxicity. Additionally, the villus width of the jejunum in the T2 treatment group decreased significantly, while all T3, T4, and T5 groups showed improvement in this regard. In summary, T. laurifolia extract can detoxify aflatoxicosis, leading to growth reduction and hepatic toxicosis in Cherry Valley ducklings.

Mycotoxins, especially aflatoxin B1 (AFB1) and fumonisin B1 (FMB1), are common contaminants in cereal-based foods. Instances of contamination are predicted to increase due to the current challenges induced by climate change. Despite the health benefits of whole grains, the presence of mycotoxins in bran remains a concern. Nonetheless, previous research indicates that wheat bran can adsorb mutagens. Therefore, this study investigated the capacity of maize, wheat, and oat brans to adsorb AFB1 and FMB1 under varying in vitro conditions, including pH, binding time, temperature, particle size, and the amount of bran utilized. Maize bran demonstrated a high AFB1 adsorption capacity (>78%) compared to wheat and oat brans. However, FMB1 was not adsorbed by the brans, possibly due to its hydrophilic nature. Lower temperature (≤25 °C) enhanced AFB1 adsorption efficacy in wheat and oat bran, while for maize bran, the highest adsorption occurred at 37 °C. A linear model following Henry’s law best explained AFB1 adsorption by the brans. Further studies identified the pericarp layer of bran as the primary site of AFB1 adsorption, with the initial liquid volume being a critical factor. The study concludes that bran could potentially act as an effective bioadsorbent. Further research is essential to confirm the adsorption efficacy and the bioavailability of AFB1 through in vivo experiments.

Aflatoxin B1 (AFB1) contamination is a food safety issue threatening human health globally. Biodegradation is an effective method for overcoming this problem, and many microorganisms have been identified as AFB1-degrading strains. However, the response mechanisms of these microbes to AFB1 remain unclear. More degrading enzymes, especially of new types, need to be discovered. In this study, a novel AFB1-degrading strain, DDC-4, was isolated using coumarin as the sole carbon source. This strain was identified as Bacillus halotolerans through physiological, biochemical, and molecular methods. The strain’s degradation activity was predominantly attributable to thermostable extracellular proteins (degradation rate remained approximately 80% at 90 °C) and was augmented by Cu2+ (95.45% AFB1 was degraded at 48 h). Alpha/beta hydrolase (arylesterase) was selected as candidate AFB1-degrading enzymes for the first time as a gene encoding this enzyme was highly expressed in the presence of AFB1. Moreover, AFB1 inhibited many genes involved in the nucleotide synthesis of strain DDC-4, which is possibly the partial molecular mechanism of AFB1’s toxicity to microorganisms. To survive under this stress, sporulation-related genes were induced in the strain. Altogether, our study identified a novel AFB1-degrading strain and explained its response mechanisms to AFB1, thereby providing new insights for AFB1 biodegradation.

Poultry may face simultaneous exposure to aflatoxin B1 (AFB1) and tiamulin (TIA), given mycotoxin contamination and antibiotic use. As both mycotoxins and antibiotics can affect cytochrome P450 enzymes (CYP450), our study aimed to explore their interaction. We developed UHPLC-MS/MS methods for the first-time determination of the interaction between TIA and AFB1 in vitro and in vivo in broiler chickens. The inhibition assay showed the half maximal inhibitory concentration (IC50) values of AFB1 and TIA in chicken liver microsomes are more than 7.6 μM, indicating an extremely weak inhibitory effect on hepatic enzymes. Nevertheless, the oral TIA pharmacokinetic results indicated that AFB1 significantly increased the area under the plasma concentration-time curve (AUClast) of TIA by 167% (p < 0.01). Additionally, the oral AFB1 pharmacokinetics revealed that TIA increased the AUClast and mean residence time (MRT) of AFB1 by 194% (p < 0.01) and 136%, respectively. These results suggested that the observed inhibition may be influenced by other factors, such as transport. Therefore, it is meaningful to further explore transport and other enzymes, involved in the interaction between AFB1 and TIA. Furthermore, additional clinical studies are necessary to thoroughly assess the safety of co-exposure with mycotoxins and antibiotics.

Seyed ahmad Sajjadi1 , Zahra Moosavi2 , Farhad Niknejad3 , Abdollah Jamshidi 4 Background: Aflatoxin B1 (AFB1) is one of the most important mycotoxins that contaminate food worldwide. Long-term consumption of foods contaminated with AFB1 endangers human health. Detoxification of AFB1 from food improves community health. A Specific approach to aflatoxin reduction is the use of probiotics. Kefir drink is a strong probiotic. The purpose of this study was to investigate the protective effect of kefir drink on AFB1-induced hepatic injury in adult male rats Methods: In this experimental study, 24 adult rats weighing between 150 and 200 g were used. The rats were randomly divided into 4 groups: 1) control, 2) AFB1 (50 μg/kg body weight), 3) kefir drink (10 mL/kg body weight), and 4) AFB1 + kefir drink. Aflatoxin and kefir drink received through oral gavage. At the end of the experiment (8 weeks), blood and liver samples were collected for different assays. Liver function tests and histopathological examinations were performed. Data were analyzed using 1-way analysis of variance (ANOVA) and at a significance level of

Introduction: Aflatoxins B1 are among the most common poisonous mycotoxins produced by certain fungi that harm animals and crops. Mycotoxins can cause a variety of adverse health effects and pose a serious health threat to humans. The Maximum Residue Limits of aflatoxin B1 in processed cereals and ingredients are 2 parts per billion (ppb) and 5 ppb, respectively. Objectives: To evaluate the status of aflatoxin B 1 contamination in rice, corn and staple food produced in Ha Giang province compared with the maximum permitted levels. Methods: A total of 210 rice and maize samples were analyzed to quantify the level of aflatoxin B1. Analysis of mycotoxins was conducted by High Performance Liquid Chromatography using a fluorescence detector. Results: It was found that rice, rice products, maize, and maize products had a mean aflatoxin B1 content of 1.79 ppb, 2.55 ppb, 2.19 ppb, and 6.35 ppb, respectively. The results also showed that 71.9% of samples were contaminated with mycotoxins, and 14.28% of samples exceeded the maximum allowable limit. Conclusion: The concentration of aflatoxin B1 in 14.28% of the samples are over permissible limits by nationwide regulations.

A recent study published data on the growth performance, relative weights of the organs of the gastrointestinal tract, liver histology, serum biochemistry, and hematological parameters for turkey poults fed an experimental diet contaminated with aflatoxin B1 (AFB1) and humic acids (HA) extracted from vermicompost. The negative effects of AFB1 (250 ng AFB1/g of feed) were significantly reduced by HA supplementation (0.25% w/w), suggesting that HA might be utilized to ameliorate the negative impact of AFB1 from contaminated diets. The present study shows the results of the remaining variables, as an extension of a previously published work which aimed to evaluate the impact of HA on the intestinal microbiota, gut integrity, ileum morphometry, and cellular immunity of turkey poults fed an AFB1-contaminated diet. For this objective, five equal groups of 1-day-old female Nicholas-700 turkey poults were randomly assigned to the following treatments: negative control (basal diet), positive control (basal diet + 250 ng AFB1/g), HA (basal diet + 0.25% HA), HA + AFB1 (basal diet + 0.25% HA + 250 ng AFB1/g), and Zeolite (basal diet + 0.25% zeolite + 250 ng AFB1/g). In the experiment, seven replicates of ten poults each were used per treatment (n = 70). In general, HA supplementation with or without the presence of AFB1 showed a significant increase (p < 0.05) in the number of beneficial butyric acid producers, ileum villi height, and ileum total area, and a significant reduction in serum levels of fluorescein isothiocyanate–dextran (FITC-d), a marker of intestinal integrity. In contrast, poults fed with AFB1 showed a significant increase in Proteobacteria and lower numbers of beneficial bacteria, clearly suggesting gut dysbacteriosis. Moreover, poults supplemented with AFB1 displayed the lowest morphometric parameters and the highest intestinal permeability. Furthermore, poults in the negative and positive control treatments had the lowest cutaneous basophil hypersensitivity response. These findings suggest that HA supplementation enhanced intestinal integrity (shape and permeability), cellular immune response, and healthier gut microbiota composition, even in the presence of dietary exposure to AFB1. These results complement those of the previously published study, suggesting that HA may be a viable dietary intervention to improve gut health and immunity in turkey poults during aflatoxicosis.

Aflatoxin B1 (AFB1), a ubiquitous mycotoxin in corn-based animal feed, particularly in tropical regions, impairs liver function, induces oxidative stress and disrupts cellular pathways, potentially worsening bone health in modern broilers. A 19-day experiment was conducted to investigate the effects of feeding increasing levels of AFB1-contaminated feed (n = 360). While growth performance remained unaffected up to Day 19, significant reductions in tibial bone ash content were observed at levels exceeding 260 ppb. Micro-computed tomography results showed that AFB1 levels at 560 ppb significantly decreased trabecular bone mineral content and density, with a tendency for reduced connectivity density in femur metaphysis. Moreover, AFB1 above 230 ppb reduced the bone volume and tissue volume of the cortical bone of femur. Even at levels above 75 ppb, AFB1 exposure significantly downregulated the jejunal mRNA expressions of the vitamin D receptor and calcium and phosphorus transporters. It can be concluded that AFB1 at levels higher than 230 ppb negatively affects bone health by impairing bone mineralization via disruption of the vitamin D receptor and calcium and phosphorus homeostasis, potentially contributing to bone health issues in broilers.

Water kefir grains (WKGs), the starter used to develop a traditional beverage named water kefir, consist of a symbiotic mixture of probiotics with diverse bioactivities, but little is known about their abilities to remove mycotoxins that have serious adverse effects on humans and animals. This study investigated the ability of WKGs to remove aflatoxin B1 (AFB1), one of the most toxic mycotoxins, under different settings, and determined the mechanism of absorption mediated by WKGs and the effect of WKGs on the toxicity induced by AFB1 and the reduction in AFB1 in cow milk and tea soups. The results showed the WKGs used herein were dominated by Lactobacillus, Acetobacter, Phenylobacterium, Sediminibacterium, Saccharomyces, Issatchenkia, and Kodamaea. HPLC analysis demonstrated that the WKGs effectively removed AFB1 at concentrations ranging from 1 to 5 µg/mL, pH values ranging from 3 to 9, and temperatures ranging from 4 to 45 °C. Additionally, the removal of AFB1 mainly depended on absorption, which was consistent with the Freundlich and pseudo-second-order kinetic models. Moreover, only 49.63% of AFB1 was released from the AFB1-WKG complex after four washes when the release of AFB1 was non-detectable. Furthermore, WKG treatment caused a dramatic reduction in the mutagenicity induced by AFB1 according to an Ames test and reduced more than 54% of AFB1 in cow milk and three tea soups. These results suggested that WKGs can act as a potential bio-absorbent with a high binding ability to detoxify AFB1 in food and feed via a chemical action step and multi-binding sites of AFB1 absorption in a wide range of scenarios.

(1) Background: Safety problems associated with aflatoxin B1 (AFB1) contamination have always been a major threat to human health. Removing AFB1 through adsorption is considered an attractive remediation technique. (2) Methods: To produce an adsorbent with a high AFB1 adsorption efficiency, a magnetic reduced graphene oxide composite (Fe3O4@rGO) was synthesized using one-step hydrothermal fabrication. Then, the adsorbent was characterized using a series of techniques, such as SEM, TEM, XRD, FT-IR, VSM, and nitrogen adsorption–desorption analysis. Finally, the effects of this nanocomposite on the nutritional components of treated foods, such as vegetable oil and peanut milk, were also examined. (3) Results: The optimal synthesis conditions for Fe3O4@rGO were determined to be 200 °C for 6 h. The synthesis temperature significantly affected the adsorption properties of the prepared material due to its effect on the layered structure of graphene and the loading of Fe3O4 nanoparticles. The results of various characterizations illustrated that the surface of Fe3O4@rGO had a two-dimensional layered nanostructure with many folds and that Fe3O4 nanoparticles were distributed uniformly on the surface of the composite material. Moreover, the results of isotherm, kinetic, and thermodynamic analyses indicated that the adsorption of AFB1 by Fe3O4@rGO conformed to the Langmuir model, with a maximum adsorption capacity of 82.64 mg·g−1; the rapid and efficient adsorption of AFB1 occurred mainly through chemical adsorption via a spontaneous endothermic process. When applied to treat vegetable oil and peanut milk, the prepared material minimized the loss of nutrients and thus preserved food quality. (4) Conclusions: The above findings reveal a promising adsorbent, Fe3O4@rGO, with favorable properties for AFB1 adsorption and potential for food safety applications.

Aflatoxins B1 (AFB1) are fungi-produced toxins found in crops like peanuts, maize, and tree nuts. They constitute a public health concern due to their genotoxic and carcinogenic effects. A deterministic exposure risk assessment to AFB1 through the consumption of peanuts was conducted on children using the Margin of Exposure (MOE) and the liver cancer risk approaches. Data on AFB1 concentrations in peanuts, quantities of peanut consumption, and the weights of the children were obtained from the literature. Generally, MOE values were below the safe margin of 10,000, ranging between 3.68 and 0.14, 754.34 and 27.33, and 11,428.57 and 419.05 for the high (0.0466 ng/kg), median (0.00023 ng/kg), and low (0.000015 ng/kg) AFB1 concentration levels, respectively. The liver cancer risk upon lifetime exposure to highly AFB1-contaminated peanuts (0.0466 ng/kg) ranged between 1 and 23 (95% lower bound) and 2 and 50 (95% upper bound) cases in a million individuals: a public health concern. A low liver cancer risk (≤1 case in a billion individuals upon lifetime exposure) was shown at median and low AFB1 concentrations. However, the risk of AFB1 should be a priority for risk management since its harmful effects could be potentiated by poor diet, high malnutrition levels, and other disease burdens in Zambia’s children.

The most frequent adverse effects of AFB1 in chicken are low performance, the depression of the immune system, and a reduced quality of both eggs and meat, leading to economic losses. Since oxidative stress plays a major role in AFB1 toxicity, natural products are increasingly being used as an alternative to mineral binders to tackle AFB1 toxicosis in farm animals. In this study, an in vivo trial was performed by exposing broilers for 10 days to AFB1 at dietary concentrations approaching the maximum limits set by the EU (0.02 mg/kg feed) in the presence or absence of turmeric powder (TP) (included in the feed at 400 mg/kg). The aims were to evaluate (i) the effects of AFB1 on lipid peroxidation, antioxidant parameters, histology, and the expression of drug transporters and biotransformation enzymes in the liver; (ii) the hepatic accumulation of AFB1 and its main metabolites (assessed using an in-house-validated HPLC-FLD method); (iii) the possible modulation of the above parameters elicited by TP. Broilers exposed to AFB1 alone displayed a significant increase in lipid peroxidation in the liver, which was completely reverted by the concomitant administration of TP. Although no changes in glutathione levels and antioxidant enzyme activities were detected in any treatment group, AFB1 significantly upregulated and downregulated the mRNA expression of CYP2A6 and Nrf2, respectively. TP counteracted such negative effects and increased the hepatic gene expression of selected antioxidant enzymes (i.e., CAT and SOD2) and drug transporters (i.e., ABCG2), which were further enhanced in combination with AFB1. Moreover, both AFB1 and TP increased the mRNA levels of ABCC2 and ABCG2 in the duodenum. The latter changes might be implicated in the decrease in hepatic AFB1 to undetectable levels (

Aflatoxin B1 (AFB1) is a hazardous mycotoxin that often contaminates animal feed and may potentially induce severe liver damage if ingested. The liver is the primary organ responsible for AFB1 detoxification through enzyme-catalyzed xenobiotic metabolism and bile acid (BA)-associated excretion. In this study, we sought to investigate whether exogenous BA improves hepatic AFB1 detoxification to alleviate AFB1-induced liver injury in broiler chickens. Five-day-old broiler chicks were randomly assigned to three groups. CON and AFB1 received a basal diet; AFB1 + BA received a basal diet with 250 mg/kg BA for 20 days. After a 3-day pre-feed, AFB1 and AFB1 + BA were daily gavaged with 250 μg/kg BW AFB1, while CON received gavage solvent for AFB1 treatment. Dietary BA supplementation protected chickens from AFB1-induced hepatic inflammation and oxidative stress. The hepatic biotransformation of AFB1 to its metabolite AFBO was improved, with accelerated excretion to the gallbladder and cecum. Accordantly, AFB1-induced down-regulation of detoxification genes, including cytochrome P450 enzymes, glutathione S-transferases, and the bile salt export pump, was rescued by BA supplementation. Moreover, liver X receptor α, suppressed by AFB1, was enhanced in BA-treated broiler chickens. These results indicate that dietary BA supplementation improves hepatic AFB1 detoxification and excretion through LXRα-involved regulation of xenobiotic enzymes.

Background and objectives: Aflatoxin B1 (AFB1) is the most toxic aflatoxin produced by a large number of Aspergillus species. Successful detoxification of this toxin is an important attempt to improve community health. The aim of this study was to evaluate reducing effects of yeasts isolates from kefir and traditional kefir-like fermented beverages on AFB1 in a broth medium. Methods: Polymerase chain reaction-sequencing was carried out to identify the yeast isolates from kefir and kefir-like beverages. Effects of the isolates on AFB1 adsorption and biotransformation in peptone dexterose broth medium were evaluated by using high performance liquid chromatography. Results: Saccharomyces cerevisiae and Kluyveromyces marxianus were isolated from kefir and kefir-like beverages and resulted in 46% and 53% AFB1 adsorption, respectively. The isolates 27Y and 2Y caused 7% toxin biotransformation, while 10% toxin biotransformation was achieved by the isolate 18Y. Conclusion: Our results indicate that the yeast isolates from kefir and traditional kefir-like products can bind to and detoxify AFB1, thereby reducing its harmful effects.

Forage grain contamination with aflatoxin B1 (AFB1) is a global problem, so its detoxification with the aim of providing feed safety and cost-efficiency is still a relevant issue. AFB1 degradation by microbial enzymes is considered to be a promising detoxification approach. In this study, we modified an previously developed Pichia pastoris GS115 expression system using a chimeric signal peptide to obtain a new recombinant producer of extracellular AFB1 oxidase (AFO) from Armillaria tabescens (the yield of 0.3 g/L), purified AFO, and selected optimal conditions for AFO-induced AFB1 removal from model solutions. After a 72 h exposure of the AFB1 solution to AFO at pH 6.0 and 30 °C, 80% of the AFB1 was degraded. Treatments with AFO also significantly reduced the AFB1 content in wheat and corn grain inoculated with Aspergillus flavus. In grain samples contaminated with several dozen micrograms of AFB1 per kg, a 48 h exposure to AFO resulted in at least double the reduction in grain contamination compared to the control, while the same treatment of more significantly (~mg/kg) AFB1-polluted samples reduced their contamination by ~40%. These findings prove the potential of the tested AFO for cereal grain decontamination and suggest that additional studies to stabilize AFO and improve its AFB1-degrading efficacy are required.

Mycotoxins are considered the most threating natural contaminants in food. Among these mycotoxins, aflatoxin B1 (AFB1) and fumonisin B1 (FB1) are the most prominent fungal metabolites that represent high food safety risks, due to their widespread co-occurrence in several food commodities, and their profound toxic effects on humans. Considering the ethical and more humane animal research, the 3Rs (replacement, reduction, and refinement) principle has been promoted in the last few years. Therefore, this review aims to summarize the research studies conducted up to date on the toxicological effects that AFB1 and FB1 can induce on human health, through the examination of a selected number of in vitro studies. Although the impact of both toxins, as well as their combination, were investigated in different cell lines, the majority of the work was carried out in hepatic cell lines, especially HepG2, owing to the contaminants’ liver toxicity. In all the reviewed studies, AFB1 and FB1 could invoke, after short-term exposure, cell apoptosis, by inducing several pathways (oxidative stress, the mitochondrial pathway, ER stress, the Fas/FasL signaling pathway, and the TNF-α signal pathway). Among these pathways, mitochondria are the primary target of both toxins. The interaction of AFB1 and FB1, whether additive, synergistic, or antagonistic, depends to great extent on FB1/AFB1 ratio. However, it is generally manifested synergistically, via the induction of oxidative stress and mitochondria dysfunction, through the expression of the Bcl-2 family and p53 proteins. Therefore, AFB1 and FB1 mixture may enhance more in vitro toxic effects, and carry a higher significant risk factor, than the individual presence of each toxin.

Aflatoxin B1 (AFB1) is one of the most toxic and harmful fungal toxins to humans and animals, and the fundamental way to prevent its entry into humans is to detect its presence in advance. In this paper, the monoclonal antibody mAbA2-2 was obtained via three-step sample amplification and multi-concentration standard detection using a subcloning method based on the limited dilution method with AFB1 as the target. A dynamic and pseucdo-homogeneous magnetic beads enzyme-linked immunosorbent assay (MBs-icELISA) was established using the prepared antibody as the recognition element and immunomagnetic beads as the antigen carrier. The MBs-icELISA showed good linear correlation in the concentration range of 0.004–10 ng/mL with R2 = 0.99396. The limit of detection (LOD) of the MBs-icELISA for AFB1 was 0.0013 ng/mL. This new ELISA strategy significantly shortened AFB1 detection time through improved sensitivity compared to the conventional ELISA method.

An adsorbent material derived from alfalfa leaves was prepared and further characterized, and its efficacy for removing aflatoxin B1 (AFB1) was investigated. Characterization consisted of the use of attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), environmental scanning electron microscopy (ESEM), X-ray fluorescence spectroscopy (XRF), X-ray diffraction (XRD), point of zero charge (pHpzc), zeta potential (ζ-potential), UV-Vis diffuse reflectance spectroscopy, and spectral analysis. To determine the adsorption capacity against AFB1 (250 ng AFB1/mL), pH-dependent and avian intestinal in vitro models were used. The adsorbent inclusion percentage was 0.5% (w/w). In general, the pH-dependent model gave adsorption percentages of 98.2%, 99.9%, and 98.2%, evaluated at pH values of 2, 5, and 7, respectively. However, when the avian intestinal model was used, it was observed that the adsorption percentage of AFB1 significantly decreased (88.8%). Based on the characterization results, it is proposed that electrostatic, non-electrostatic, and the formation of chlorophyll-AFB1 complexes were the main mechanisms for AFB1 adsorption. From these results, it can be concluded that the adsorbent derived from alfalfa leaves could be used as an effective material for removing AFB1 in in vitro digestion models that mimic the physiological reality.

Mycotoxin contamination on food and feed can have deleterious effect on human and animal health. Agricultural crops may contain one or more mycotoxin compounds; therefore, a good multiplex detection method is desirable to ensure food safety. In this study, we developed a rapid method using label-free surface-enhanced Raman spectroscopy (SERS) to simultaneously detect three common types of mycotoxins found on corn, namely aflatoxin B1 (AFB1), zearalenone (ZEN), and ochratoxin A (OTA). The intrinsic chemical fingerprint from each mycotoxin was characterized by their unique Raman spectra, enabling clear discrimination between them. The limit of detection (LOD) of AFB1, ZEN, and OTA on corn were 10 ppb (32 nM), 20 ppb (64 nM), and 100 ppb (248 nM), respectively. Multivariate statistical analysis was used to predict concentrations of AFB1, ZEN, and OTA up to 1.5 ppm (4.8 µM) based on the SERS spectra of known concentrations, resulting in a correlation coefficient of 0.74, 0.89, and 0.72, respectively. The sampling time was less than 30 min per sample. The application of label-free SERS and multivariate analysis is a promising method for rapid and simultaneous detection of mycotoxins in corn and may be extended to other types of mycotoxins and crops.

Aflatoxin B1 (AFB1) induces lipid peroxidation and mortality in bovine foetal hepatocyte-derived cells (BFH12), with underlying transcriptional perturbations associated mainly with cancer, cellular damage, inflammation, bioactivation, and detoxification pathways. In this cell line, curcumin and resveratrol have proven to be effective in mitigating AFB1-induced toxicity. In this paper, we preliminarily assessed the potential anti-AFB1 activity of a natural polyphenol, quercetin (QUE), in BFH12 cells. To this end, we primarily measured QUE cytotoxicity using a WST-1 reagent. Then, we pre-treated the cells with QUE and exposed them to AFB1. The protective role of QUE was evaluated by measuring cytotoxicity, transcriptional changes (RNA-sequencing), lipid peroxidation (malondialdehyde production), and targeted post-transcriptional modifications (NQO1 and CYP3A enzymatic activity). The results demonstrated that QUE, like curcumin and resveratrol, reduced AFB1-induced cytotoxicity and lipid peroxidation and caused larger transcriptional variations than AFB1 alone. Most of the differentially expressed genes were involved in lipid homeostasis, inflammatory and immune processes, and carcinogenesis. As for enzymatic activities, QUE significantly reverted CYP3A variations induced by AFB1, but not those of NQO1. This study provides new knowledge about key molecular mechanisms involved in QUE-mediated protection against AFB1 toxicity and encourages in vivo studies to assess QUE’s bioavailability and beneficial effects on aflatoxicosis.

Fungal infection and mycotoxin contamination are major hazards to the safe storage and distribution of foods and feeds consumed by humans and livestock. This study investigated the antifungal and antiaflatoxigenic activities of massoia essential oil (MEO) and its major constituent, C10 massoia lactone (C10), against aflatoxin B (AFB)-producing Aspergillus flavus ATCC 22546. Their antifungal activities were evaluated using a disc diffusion assay, agar dilution method, and a mycelial growth inhibition assay with the AFB analysis using liquid chromatography triple quadrupole mass spectrometry. MEO and C10 exhibited similar antifungal and antiaflatoxigenic activities against A. flavus. C10 was a primary constituent in MEO and represented up to 45.1% of total peak areas analyzed by gas chromatography–mass spectrometry, indicating that C10 is a major compound contributing to the antifungal and antiaflatoxigenic activities of MEO. Interestingly, these two materials increased AFB production in A. flavus by upregulating the expression of most genes related to AFB biosynthesis by 3- to 60-fold. Overall, MEO and C10 could be suitable candidates as natural preservatives to control fungal infection and mycotoxin contamination in foods and feeds as Generally Recognized As Safe (GRAS) in the Flavor and Extract Manufacturers Association of the United States (FEMA), and MEO is a more suitable substance than C10 because of its wider range of uses and higher allowed concentration than C10.

Maize is frequently contaminated with multiple mycotoxins, especially those produced by Aspergillus flavus and Fusarium verticillioides. As mycotoxin contamination is a critical factor that destabilizes global food safety, the current review provides an updated overview of the (co-)occurrence of A. flavus and F. verticillioides and (co-)contamination of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) in maize. Furthermore, it summarizes their interactions in maize. The gathered data predict the (co-)occurrence and virulence of A. flavus and F. verticillioides would increase worldwide, especially in European cold climate countries. Studies on the interaction of both fungi regarding their growth mainly showed antagonistic interactions in vitro or in planta conditions. However, the (co-)contamination of AFB1 and FB1 has risen worldwide in the last decade. Primarily, this co-contamination increased by 32% in Europe (2010–2020 vs. 1992–2009). This implies that fungi and mycotoxins would severely threaten European-grown maize.

Aflatoxins can cause intoxication and poisoning in animals and humans. Among these molecules, aflatoxin B1 (AFB1) is the most dangerous because of its carcinogenic and mutagenic properties. To mitigate these effects, clay adsorbents are commonly included in the diet of animals to adsorb the carcinogens and prevent their absorption in the gastrointestinal tract. In this study, four clays, three smectites (C-1, C-2, and C-3), and one zeolite (C-4), were compared as adsorbents of AFB1 and trace inorganic nutrients using an in vitro gastrointestinal model for poultry. Characterization of the clays using Fourier transform infrared spectroscopy revealed characteristic bands of smectites in C-1, C-2, and C-3 (stretching vibrations of Si-O, Al-O-Si, and Si-O-Si). The C-4 presented bands related to the bending vibration of structural units (Si-O-Si and Al-O-Si). X-ray diffraction analysis showed that C-1 is a montmorillonite, C-2 is a beidellite, C-3 is a beidellite-Ca-montmorillonite, and C-4 is a clinoptilolite. The elemental compositions of the clays showed alumina, silica, iron, calcium, and sodium contents. The cation exchange capacity was higher in C-3 clay (60.2 cmol(+)/kg) in contrast with the other clays. The AFB1 adsorption of C-1 was the highest (98%; p ˂ 0.001), followed by C-2 (94%). However, all the clays also sequestered trace inorganic nutrients (Fe, Mn, Zn, and Se). Both smectites, montmorillonite and beidellite, were the most suitable for use as adsorbents of AFB1.

Mycotoxins are natural food and feed contaminants produced by several molds. The primary mode of exposure in humans and animals is through mixtures. Aflatoxin B1 (AFB1) and sterigmatocystin (STER) are structurally related mycotoxins that share the same biosynthetic route. Few in vivo genotoxicity assays have been performed with STER. In the present genotoxicity study, Wistar rats were dosed orally with STER (20 mg/kg b.w.), AFB1 (0.25 mg/kg b.w.) or a mixture of both in an integrated micronucleus (bone marrow) and comet study (liver and kidney). STER was dosed at the highest feasible dose in corn oil. No increase in the percentage of micronuclei in bone marrow was observed at any condition. Slight DNA damage was detected in the livers of animals treated with AFB1 or the mixture (DNA strand breaks and Fpg (Formamidopyrimidine DNA glycosylase)-sensitive sites, respectively). Plasma, liver, and kidney samples were analyzed with LC-MS/MS demonstrating exposure to both mycotoxins. General toxicity parameters (organs absolute weight, biochemistry, and histopathology) were not altered either individually or in the mixture. The overall absence of individual genotoxicity did not allow us to set any type of interaction in the mixture. However, a possible toxicokinetic interaction was observed.

Crop contamination by aflatoxin B1 (AFB1), an Aspergillus-flavus-produced toxin, is frequently observed in tropical and subtropical regions. This phenomenon is emerging in Europe, most likely as a result of climate change. Alternative methods, such as biocontrol agents (BCAs), are currently being developed to reduce the use of chemicals in the prevention of mycotoxin contamination. Actinobacteria are known to produce many bioactive compounds, and some of them can reduce in vitro AFB1 concentration. In this context, the present study aims to analyze the effect of a cell-free supernatant (CFS) from Streptomyces roseolus culture on the development of A. flavus, as well as on its transcriptome profile using microarray assay and its impact on AFB1 concentration. Results demonstrated that in vitro, the S. roseolus CFS reduced the dry weight and conidiation of A. flavus from 77% and 43%, respectively, and was therefore associated with a reduction in AFB1 concentration reduction to levels under the limit of quantification. The transcriptomic data analysis revealed that 5198 genes were differentially expressed in response to the CFS exposure and among them 5169 were downregulated including most of the genes involved in biosynthetic gene clusters. The aflatoxins’ gene cluster was the most downregulated. Other gene clusters, such as the aspergillic acid, aspirochlorine, and ustiloxin B gene clusters, were also downregulated and associated with a variation in their concentration, confirmed by LC-HRMS.

Phenyllactic acid (PLA), a promising food preservative, is safe and effective against a broad spectrum of food-borne pathogens. However, its mechanisms against toxigenic fungi are still poorly understood. In this study, we applied physicochemical, morphological, metabolomics, and transcriptomics analyses to investigate the activity and mechanism of PLA inhibition of a typical food-contaminating mold, Aspergillus flavus. The results showed that PLA effectively inhibited the growth of A. flavus spores and reduced aflatoxin B1 (AFB1) production by downregulating key genes associated with AFB1 biosynthesis. Propidium iodide staining and transmission electron microscopy analysis demonstrated a dose-dependent disruption of the integrity and morphology of the A. flavus spore cell membrane by PLA. Multi-omics analyses showed that subinhibitory concentrations of PLA induced significant changes in A. flavus spores at the transcriptional and metabolic levels, as 980 genes and 30 metabolites were differentially expressed. Moreover, KEGG pathway enrichment analysis indicated PLA-induced cell membrane damage, energy-metabolism disruption, and central-dogma abnormality in A. flavus spores. The results provided new insights into the anti-A. flavus and -AFB1 mechanisms of PLA.

Fungal contamination of animal feed can severely affect the health of farm animals, and result in considerable economic losses. Certain filamentous fungi or molds produce toxic secondary metabolites known as mycotoxins, of which aflatoxins (AFTs) are considered the most critical dietary risk factor for both humans and animals. AFTs are ubiquitous in the environment, soil, and food crops, and aflatoxin B1(AFB1) has been identified by the World Health Organization (WHO) as one of the most potent natural group 1A carcinogen. We reviewed the literature on the toxic effects of AFB1 in humans and animals along with its toxicokinetic properties. The damage induced by AFB1 in cells and tissues is mainly achieved through cell cycle arrest and inhibition of cell proliferation, and the induction of apoptosis, oxidative stress, endoplasmic reticulum (ER) stress and autophagy. In addition, numerous coding genes and non-coding RNAs have been identified that regulate AFB1 toxicity. This review is a summary of the current research on the complexity of AFB1 toxicity, and provides insights into the molecular mechanisms as well as the phenotypic characteristics.

Aflatoxins (AFs) are toxic secondary metabolites produced by Aspergillus spp. and are found in food and feed as contaminants worldwide. Due to climate change, AFs occurrence is expected to increase also in western Europe. Therefore, to ensure food and feed safety, it is mandatory to develop green technologies for AFs reduction in contaminated matrices. With this regard, enzymatic degradation is an effective and environmentally friendly approach under mild operational conditions and with minor impact on the food and feed matrix. In this work, Ery4 laccase, acetosyringone, ascorbic acid, and dehydroascorbic acid were investigated in vitro, then applied in artificially contaminated corn for AFB1 reduction. AFB1 (0.1 µg/mL) was completely removed in vitro and reduced by 26% in corn. Several degradation products were detected in vitro by UHPLC-HRMS and likely corresponded to AFQ1, epi-AFQ1, AFB1-diol, or AFB1dialehyde, AFB2a, and AFM1. Protein content was not altered by the enzymatic treatment, while slightly higher levels of lipid peroxidation and H2O2 were detected. Although further studies are needed to improve AFB1 reduction and reduce the impact of this treatment in corn, the results of this study are promising and suggest that Ery4 laccase can be effectively applied for the reduction in AFB1 in corn.

Rice is the second most important cereal crop and is vital for the diet of billions of people. However, its consumption can increase human exposure to chemical contaminants, namely mycotoxins and metalloids. Our goal was to evaluate the occurrence and human exposure of aflatoxin B1 (AFB1), ochratoxin A (OTA), zearalenone (ZEN), and inorganic arsenic (InAs) in 36 rice samples produced and commercialized in Portugal and evaluate their correlation. The analysis of mycotoxins involved ELISA, with limits of detection (LODs) of 0.8, 1 and 1.75 μg kg−1 for OTA, AFB1, and ZEN, respectively. InAs analysis was carried out by inductively coupled plasma mass spectrometry (ICP-MS; LOD = 3.3 μg kg−1). No sample showed contamination by OTA. AFB1 was present in 2 (4.8%) samples (1.96 and 2.20 μg kg−1), doubling the European maximum permitted level (MPL). Concerning ZEN, 88.89% of the rice samples presented levels above the LOD up to 14.25 µg kg−1 (average of 2.75 µg kg−1). Regarding InAs, every sample presented concentration values above the LOD up to 100.0 µg kg−1 (average of 35.3 µg kg−1), although none surpassed the MPL (200 µg kg−1). No correlation was observed between mycotoxins and InAs contamination. As for human exposure, only AFB1 surpassed the provisional maximum tolerable daily intake. Children were recognized as the most susceptible group.

Aflatoxin B1 (AFB1) and aflatoxin M1 (AFM1) are universally found as environmental pollutants. AFB1 and AFM1 are group 1 human carcinogens. Previous sufficient toxicological data show that they pose a health risk. The intestine is vital for resistance to foreign pollutants. The enterotoxic mechanisms of AFB1 and AFM1 have not been clarified at the metabolism levels. In the present study, cytotoxicity evaluations of AFB1 and AFM1 were conducted in NCM 460 cells by obtaining their half-maximal inhibitory concentration (IC50). The toxic effects of 2.5 μM AFB1 and AFM1 were determined by comprehensive metabolomics and lipidomics analyses on NCM460 cells. A combination of AFB1 and AFM1 induced more extensive metabolic disturbances in NCM460 cells than either aflatoxin alone. AFB1 exerted a greater effect in the combination group. Metabolomics pathway analysis showed that glycerophospholipid metabolism, fatty acid degradation, and propanoate metabolism were dominant pathways that were interfered with by AFB1, AFM1, and AFB1+AFM1. Those results suggest that attention should be paid to lipid metabolism after AFB1 and AFM1 exposure. Further, lipidomics was used to explore the fluctuation of AFB1 and AFM1 in lipid metabolism. The 34 specific lipids that were differentially induced by AFB1 were mainly attributed to 14 species, of which cardiolipin (CL) and triacylglycerol (TAG) accounted for 41%. AFM1 mainly affected CL and phosphatidylglycerol, approximately 70% based on 11 specific lipids, while 30 specific lipids were found in AFB1+AFM1, mainly reflected in TAG up to 77%. This research found for the first time that the lipid metabolism disorder caused by AFB1 and AFM1 was one of the main causes contributing to enterotoxicity, which could provide new insights into the toxic mechanisms of AFB1 and AFM1 in animals and humans.

Aflatoxin B1 (AFB1), ochratoxin A (OTA), and deoxynivalenol (DON) are the three mycotoxins that have received the most scholarly attention and have been tested most routinely in clinics. These mycotoxins not only suppress immune responses but also induce inflammation and even increase susceptibility to pathogens. Here, we comprehensively reviewed the determining factors for the bidirectional immunotoxicity of the three mycotoxins, their effects on pathogens, and their action mechanisms. The determining factors include mycotoxin exposure doses and times, as well as species, sex, and some immunologic stimulants. Moreover, mycotoxin exposure can affect the infection severity of some pathogens, including bacteria, viruses, and parasites. Their specific action mechanisms include three aspects: (1) mycotoxin exposure directly promotes the proliferation of pathogenic microorganisms; (2) mycotoxins produce toxicity, destroy the integrity of the mucosal barrier, and promote inflammatory response, thereby improving the susceptibility of the host; (3) mycotoxins reduce the activity of some specific immune cells and induce immune suppression, resulting in reduced host resistance. The present review will provide a scientific basis for the control of these three mycotoxins and also provide a reference for research on the causes of increased subclinical infections.

The toxins produced by Aspergillus flavus can significantly inhibit the use of maize. As a result of climate change, toxin production is a problem not only in tropical and subtropical areas but in an increasing number of European countries, including Hungary. The effect of meteorological factors and irrigation on mould colonization and aflatoxin B1 (AFB1) mycotoxin production by A. flavus were investigated in natural conditions, as well as the inoculation with a toxigenic isolate in a complex field experiment for three years. As a result of irrigation, the occurrence of fungi increased, and toxin production decreased. The mould count of fungi and toxin accumulation showed differences during the examined growing seasons. The highest AFB1 content was found in 2021. The main environmental factors in predicting mould count were temperature (Tavg, Tmax ≥ 30 °C, Tmax ≥ 32 °C, Tmax ≥ 35 °C) and atmospheric drought (RHmin ≤ 40%). Toxin production was determined by extremely high daily maximum temperatures (Tmax ≥ 35 °C). At natural contamination, the effect of Tmax ≥ 35 °C on AFB1 was maximal (r = 0.560–0.569) in the R4 stage. In the case of artificial inoculation, correlations with environmental factors were stronger (r = 0.665–0.834) during the R2–R6 stages.

Aflatoxin B1 (AFB1), a typical fungal toxin found in feed, is highly carcinogenic. Oxidative stress is one of the main ways it exerts its toxicity; therefore, finding a suitable antioxidant is the key to reducing its toxicity. Astaxanthin (AST) is a carotenoid with strong antioxidant properties. The aim of the present research was to determine whether AST eases the AFB1-induced impairment in IPEC-J2 cells, and its specific mechanism of action. AFB1 and AST were applied to IPEC-J2 cells in different concentrations for 24 h. The AST (80 µM) significantly prevented the reduction in the IPEC-J2 cell viability that was induced by AFB1 (10 μM). The results showed that treatment with AST attenuated the AFB1-induced ROS, and cytochrome C, the Bax/Bcl2 ratio, Caspase-9, and Caspase-3, which were all activated by AFB1, were among the pro-apoptotic proteins which were diminished by AST. AST activates the Nrf2 signaling pathway and ameliorates antioxidant ability. This was further evidenced by the expression of the HO-1, NQO1, SOD2, and HSP70 genes were all upregulated. Taken together, the findings show that the impairment of oxidative stress and apoptosis, caused by the AFB1 in the IPEC-J2 cells, can be attenuated by AST triggering the Nrf2 signaling pathway.

Background and Aim: Aflatoxin B1 is a common contaminant of poultry feed and has the highest rate of acute and chronic toxicity among all mycotoxins. Over the past decade, studies have well established the negative effects of aflatoxin B1 on the health of poultry. Therefore, it is necessary to determine the maximum concentration of aflatoxin B1 in poultry feed. Materials and Methods: In this study, in order to determine the aflatoxin B1 concentration in poultry feed, 45 samples were collected via a simple random method during autumn 2018 and winter 2019, and then the concentration of this contaminant was measured by the ELISA method. Results: The experimental results showed that the contamination range in the positive samples was between 6.44 and 34.18 µg/Kg. Moreover, the mean concentration of aflatoxin in contaminated samples was 5.10 µg/Kg. The concentration of contaminant in 46.66% of the samples exceeded the standard limit. Due to higher temperatures and suitable environmental conditions for the growth of fungi, the samples prepared in autumn were more contaminated than those prepared in winter. Conclusion: The level of aflatoxin B1 contamination in poultry feed in Sanandaj is high. In order to prevent the entry of this toxin into the human food cycle regular survey and monitoring of poultry feed by health institutions are necessary.

Aflatoxin B1 (AFB1) is a secondary metabolite produced by the fungus Aspergillus, which is ubiquitous in moldy grain products. Aflatoxin B1 has been reported to possess hepatotoxicity, renal toxicity, and reproductive toxicity. Previous studies have shown that AFB1 is toxic to mammalian oocytes. However, the potential toxicity of AFB1 on the organelles of mouse oocytes is unknown. In this study, we found that exposure to AFB1 significantly reduced mouse oocyte development capacity. Further analysis showed that the endoplasmic reticulum (ER) failed to accumulate around the spindle, and scattered in the cytoplasm under AFB1 exposure. Similar to the ER, the Golgi apparatus showed a uniform localization pattern following AFB1 treatment. In addition, we found that AFB1 exposure caused the condensation of lysosomes in the cytoplasm, presenting as a clustered or spindle peripheral-localization pattern, which indicated that protein modification, transport, and degradation were affected. Mitochondrial distribution was also altered by AFB1 treatment. In summary, our study showed that AFB1 exposure had toxic effects on the distribution of mouse oocyte organelles, which further led to a decline in oocyte quality.

Mycotoxins, which are produced from fungi, have harmful effects on human and animals. It has been reported in several in vivo and in vitro studies that mycotoxins caused disorders on digestive, urinary, immune, reproduction and nervous systems, however detailed molecular mechanism of toxicity needed to clarify. Mycotoxins could have potential to inducing neurotoxicity. In this study we aimed to evaluate mycotoxin aflatoxin B1(AFB1) and Fumonisin B1s (FB1) effects on cellular stress conditions and endoplasmic reticulum stress molecules gene expressions. Cell survival analysis was conducted by MTT assay. Total-ROS analysis was performed via DC-FDA analysis with fluorescent microplate reader. IRE1 alpha and Gadd 153 which are endoplasmic reticulum stress molecules gene expressions were evaluated by quantitative (real‑time) polymerase chain reaction (qPCR). According to MTT results both AFB1 and FB1 were shown 90% cell survival at 12,5-100µM concentrations range for 24h in SH-SY5Y cell line. Both 100 and 50 µM concentrations of AFB1 and FB1 were significantly induced ROS formation in SH-SY5Y cells (p [Med-Science 2020; 9(2.000): 492-5]

Two of the mycotoxins of greatest agroeconomic significance are aflatoxin B1 (AFB1), and ochratoxin A (OTA). It has been reported that extracts from some wood-decaying mushrooms, such as Lentinula edodes and Trametes versicolor showed the ability to inhibit AFB1 or OTA biosynthesis. Therefore, in our study, a wide screening of 42 isolates of different ligninolytic mushrooms was assayed for their ability to inhibit the synthesis of OTA in Aspergillus carbonarius and AFB1 in Aspergillus flavus, in order to find a metabolite that can simultaneously inhibit both mycotoxins. The results showed that four isolates produce metabolites able to inhibit the synthesis of OTA, and 11 isolates produced metabolites that inhibited AFB1 by >50%. Two strains, the Trametes versicolor strain TV117 and the Schizophyllum commune strain S.C. Ailanto, produced metabolites able to significantly inhibit (>90%) the synthesis of both mycotoxins. Preliminary results suggest that the mechanism of efficacy of the S. commune rough and semipurified polysaccharides could be analogous to that found previously for Tramesan®, by enhancing the antioxidant response in the target fungal cells. The overall results indicate that S. commune’s polysaccharide(s) could be a potential agent(s) in biological control and/or a useful component of the integrated strategies able to control mycotoxin synthesis.

The degradation of aflatoxin (AF) is a topic that always exists along with the food and feed industry. Photocatalytic degradation as an advanced oxidation technology has many benefits, including complete inorganic degradation, no secondary contamination, ease of activity under moderate conditions, and low cost compared with traditional physical, chemical, and biological strategies. However, photocatalysts are usually dispersed during photocatalytic reactions, resulting in energy and time consumption in the separation process. There is even a potential secondary pollution problem from the perspective of food safety. In this regard, three electrospun membranes anchored with g-C3N4/MoS2 composites were prepared for highly efficient photocatalytic degradation of aflatoxin B1 (AFB1) under visible light. These photocatalytic membranes were characterized by XRD, SEM, TEM, FTIR, and XPS. The factors influencing the degradation efficiency of AFB1, including pH values and initial concentrations, were also probed. The three kinds of photocatalytic membranes all exhibited excellent ability to degrade AFB1. Among them, the photocatalytic degradation efficiency of the photocatalytic membranes prepared by the coaxial methods reached 96.8%. The experiment is with an initial concentration of 0.5 μg/mL (500 PPb) after 60 min under visible light irradiation. The mechanism of degradation of AFB1 was also proposed based on active species trapping experiments. Moreover, the prepared photocatalytic membranes exhibited excellent photocatalytic activity even after five-fold use in the degradation of AFB1. These studies showed that electrospun membranes anchored with g-C3N4/MoS2 composites have a high photocatalytic ability which is easily removed from the reacted medium for reuse. Thereby, our study offers a highly effective, economical, and green solution for AFB1 degradation in the foodstuff for practical application.

Vermicompost was used for humic acid (HA) preparation, and the adsorption of aflatoxin B1 (AFB1) was investigated. Two forms of HA were evaluated, natural HA and sodium-free HA (SFHA). As a reference, a non-commercial zeolitic material was employed. The adsorbents were characterized by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), energy-dispersive X-ray spectroscopy (EDS), zeta potential (ζ-potential), scanning electron microscopy (SEM), and point of zero charge (pHpzc). The adsorbent capacity of the materials when added to an AFB1-contaminated diet (100 µg AFB1/kg) was evaluated using an in vitro model that simulates the digestive tract of chickens. Characterization results revealed the primary functional groups in HA and SFHA were carboxyl and phenol. Furthermore, adsorbents have a highly negative ζ-potential at the three simulated pH values. Therefore, it appears the main influencing factors for AFB1 adsorption are electrostatic interactions and hydrogen bonding. Moreover, the bioavailability of AFB1 in the intestinal section was dramatically decreased when sorbents were added to the diet (0.2%, w/w). The highest AFB1 adsorption percentages using HA and SFHA were 97.6% and 99.7%, respectively. The zeolitic material had a considerable adsorption (81.5%). From these results, it can be concluded that HA and SFHA from vermicompost could be used as potential adsorbents to remove AFB1 from contaminated feeds.

Aflatoxin B1 (AFB1) is a mycotoxin known to impair human and animal health. It is also believed to have a deleterious effect on ruminal nutrient digestibility under in vitro batch culture systems. The objective of this study was to evaluate the effects of increasing the dose of AFB1 on ruminal dry matter and nutrient digestibility, fermentation profile, and N flows using a dual-flow continuous culture system fed a diet formulated for lactating dairy cows. Eight fermenter vessels were used in a replicated 4 × 4 Latin square design with 10 d periods (7 d adaptation and 3 d sample collection). Treatments were randomly applied to fermenters on diet DM basis: (1) 0 μg of AFB1/kg of DM (Control); (2) 50 μg of AFB1/kg of DM (AF50); (3) 100 μg of AFB1/kg of DM (AF100); and (4) 150 μg of AFB1/kg of DM (AF150). Treatments did not affect nutrient digestibility, fermentation, and N flows. Aflatoxin B1 concentration in ruminal fluid increased with dose but decreased to undetectable levels after 4 h post-dosing. In conclusion, adding incremental doses of AFB1 did not affect ruminal fermentation, digestibility of nutrients, and N flows in a dual-flow continuous culture system fed diets formulated for lactating dairy cows.

Aflatoxin is a potent mycotoxin and a common source of grain contamination that leads to great economic losses and health problems. Although distilled baijiu cannot be contaminated by aflatoxin, its presence in the brewing process affects the physiological activities of micro-organisms and reduces product quality. Bacillus cereus XSWW9 capable of degrading aflatoxin B1 (AFB1) was isolated from daqu using coumarin as the sole carbon source. XSWW9 degraded 86.7% of 1 mg/L AFB1 after incubation at 37 °C for 72 h and tolerated up to 1 mg/L AFB1 with no inhibitory effects. Enzymes in the cell-free supernatant of XSSW9 played a significant role in AFB1 degradation. The AFB1-degradation activity was sensitive to protease K and SDS treatment, which indicated that extracellular proteins were responsible for the degradation of AFB1. In order to investigate the AFB1-degradation ability of XSSW9 during the baijiu brewing process, AFB1 and XSWW9 were added to grain fermentation (FG-T) and normal grain fermentation without AFB1, while normal grain fermentation without AFB1 and XSWW9 was used as a control (FG-C). At the end of the fermentation, 99% AFB1 was degraded in the residue of fermented grains. The differences of microbial communities in the fermented grains showed that there were no significant differences between FG-T and FG-C in the relative abundance of dominant genera. The analysis of volatile compounds of their distillation showed that the contents of skeleton flavor components was similar between FG-T and FG-C. These results offer a basis for the development of effective strategies to reduce the effect of AFB1 on the brewing process and ensure that the production of baijiu is stable.

Aflatoxin B1 (AFB1), one of the most common environmental mycotoxin contaminations in food and feed, poses significant threats to human and animal health. Our previous study indicated that even non-toxic AFB1 concentrations could promote influenza virus replication and induce influenza virus-infected alveolar macrophages polarizing from M1 (immunostimulatory phenotype) to M2 (immunosuppressive phenotype) over time. However, whether AFB1 promotes influenza replication via modulating the polarization of alveolar macrophages is unknown. Here, we specifically depleted alveolar macrophages using clodronate-containing liposomes in swine influenza virus (SIV)-infected mice to explore the mechanism the promotion of SIV replication by AFB1. The results show that the depletion of alveolar macrophages significantly alleviated the AFB1-induced weight loss, inflammatory responses, and lung and immune organ damage of the SIV-infected mice after 14 days and greatly diminished the AFB1-promoted SIV replication. In contrast, the depletion of alveolar macrophages did not alleviate the AFB1-induced weight loss, and lung and immune organ damage of the SIV-infected mice after 28 days and slightly diminished the AFB1-promoted SIV replication. Collectively, the data indicate that alveolar macrophages play a crucial role the promotion of SIV infection by AFB1 in the early rather than late stage, and AFB1 can promote SIV replication by inducing alveolar macrophages to polarize towards M1 macrophages. This research provides novel targets for reducing the risk of AFB1-promoted influenza virus infection.

Aflatoxin B1 (AFB1) exists widely in feed and food with severe hazards, posing a serious threat to human and animal health. Epigallocatechin gallate (EGCG) and glutathione (GSH) have been reported as having anti-oxidative and other functions. The present study aimed to investigate the detoxification effect of EGCG and GSH alone or in combination on AFB1 exposure in ducklings. Fifty one-day-old male ducklings were randomly assigned into five experimental groups (n = 10): 1. Control (CTR); 2. 0.3 mg/kg BW AFB1 (AFB1); 3. 0.3 mg/kg BW AFB1 + 100 mg/kg BW EGCG (AFB1 + EGCG); 4. 0.3 mg/kg BW AFB1 + 30 mg/kg BW GSH (AFB1 + GSH); 5. 0.3 mg/kg BW AFB1 + 100 mg/kg BW EGCG + 30 mg/kg BW GSH (AFB1 + EGCG + GSH). The experiment lasted for seven days. Compared with the CTR group, AFB1 reduced growth performance, total serum protein and albumin content, increased serum enzyme activity (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and γ-glutamyl transpeptidase), and caused pathological damage to the ducklings’ livers. AFB1 exposure increased malondialdehyde content and decreased superoxide dismutase, total antioxidant capacity, catalase, glutathione peroxidase activities, and glutathione content in the liver. EGCG and GSH alone or in combination mitigated these adverse effects. Meanwhile, EGCG and GSH attenuate apoptosis of hepatocytes, and regulated AFB1-induced changes in the abundance of genes contained in the Keap1/Nrf2 signalling and apoptotic pathways. Collectively, these results suggest that EGCG and GSH alleviate the hepatocyte injury induced by AFB1 by inhibiting oxidative stress and attenuating excessive mitochondria-mediated apoptosis.

The widespread fungal toxin Aflatoxin B1 (AFB1) is an inevitable pollutant affecting the health of humans, poultry, and livestock. Although studies indicate that AFB1 is hepatotoxic, there are few studies on AFB1-induced hepatotoxicity in sheep. Thus, this study examined how AFB1 affected sheep liver function 24 h after the animals received 1 mg/kg bw of AFB1 orally (dissolved in 20 mL, 4% v/v ethanol). The acute AFB1 poisoning caused histopathological injuries to the liver and increased total bilirubin (TBIL) and alkaline phosphatase (AKP) levels. AFB1 also markedly elevated the levels of the pro-inflammatory cytokines TNF-α and IL-6 while considerably reducing the expression of antioxidation-related genes (SOD-1 and SOD-2) and the anti-inflammatory gene IL-10 in the liver. Additionally, it caused apoptosis by dramatically altering the expression of genes associated with apoptosis including Bax, Caspase-3, and Bcl-2/Bax. Notably, AFB1 exposure altered the gut microbiota composition, mainly manifested by BF311 spp. and Alistipes spp. abundance, which are associated with liver injury. In conclusion, AFB1 can cause liver injury and liver dysfunction in sheep via oxidative stress, inflammation, apoptosis, and gut-microbiota disturbance.

Aspergillus flavus produces carcinogenic and mutagenic aflatoxins, which cause economic losses and risk of food safety by contaminating grains, food and feed. In this study, we characterized two bZIP transcription factors, AflatfA and AflatfB, and their genetic interaction. Compared to the wild type (WT), AflatfA deletion and AflatfA and AflatfB double deletion both caused retarded vegetative growth of mycelia. Relative to WT, the AflatfA deletion strain (ΔAflatfA) and AflatfA and AflatfB double deletion strain (ΔAflatfAΔAflatfB) produced more sclerotia, whereas the AflatfB deletion strain (ΔAflatfB) produced less sclerotia. After 4 °C preservation and incubation at 50 °C, conidia viability dramatically decreased in the ΔAflatfA and ΔAflatfAΔAflatfB but ΔAflatfB mutants, whereas conidia viability of the ΔAflatfAΔAflatfB strain was higher after storage at 4 °C than in AflatfA mutant. Conidia of ΔAflatfA, ΔAflatfB and ΔAflatfAΔAflatfB strains significantly increased in sensitivity to H2O2 in comparison with WT. Compared to WT, the mycelium of ΔAflatfA and ΔAflatfB strains were more sensitive to H2O2; conversely, the ΔAflatfAΔAflatfB strain showed less sensitivity to H2O2. ΔAflatfA and ΔAflatfAΔAflatfB strains displayed less sensitivity to the osmotic reagents NaCl, KCl and Sorbitol, in comparison with WT and ΔAflatfB strains. When on YES medium and hosts corn and peanut, ΔAflatfA and ΔAflatfAΔAflatfB strains produced less aflatoxin B1 (AFB1) than ΔAflatfB, and the AFB1 yield of ΔAflatfB was higher than that of WT. When WT and mutants were inoculated on corn and peanut, the ΔAflatfA and ΔAflatfAΔAflatfB but not ΔAflatfB mutants produced less conidia than did WT. Taken together, this study reveals that AflatfA controls more cellular processes, and the function of AflatfA is stronger than that of AflatfB when of the same process is regulated, except the response to H2O2, which might result from the effect of AflatfA on the transcriptional level of AflatfB.

The aim of the present study was to develop meat-based media with compositions similar to those of dry-fermented meat products and to evaluate their use in studying the growth of Aspergillus parasiticus and the kinetics of aflatoxin B1 (AFB1) production. In our previous experiments, we found that the strain A. parasiticus ŽMJ7 produced a high amount of AFB1. Cooked meat agar (CMA2) was used as a novel complex meat-based medium with four variations: CMA2G (CMA2 supplemented with 1% glucose), CMA2YE (CMA2 supplemented with 0.2% yeast extract), and CMA2GYE (CMA2 supplemented with 1% glucose and 0.2% yeast extract). Media were inoculated with an A. parasiticus spore suspension (105 spores/mL) and incubated at 25 °C for up to 15 days. The A. parasiticus lag phase lasted less than 1 day, irrespective of the growth medium, with the exception of control medium CMA1 (cooked meat agar) as an already known meat-based medium. The highest mean colony growth rate was observed on CMA2 and CMA2G. Reversed-phase UPLC–MS/MS analysis was performed to determine the AFB1 concentration in combination with solid phase extraction (SPE). The highest AFB1 concentration in meat-based media was detected in CMA2GYE after 15 days of incubation (13,502 ± 2367 ng/mL media). The results showed that for studying AFB1 production in dry-fermented meat products, novel suitable media such as CMA2-based media are required. This finding could represent a potential concern with regard to the production of dry-fermented meat products.

The degradation of aflatoxin B1 (AFB1) and zearalenone (ZEA) is investigated using power ultrasound to identify suitable methods to reduce the mycotoxin content of corn. AFB1 and ZEA in corn are simultaneously degraded via power ultrasound; thus, this method has a significant effect on corn quality. The power intensity, solid-liquid ratio, and ultrasonic treatment modes significantly affect the degradation rates of AFB1 and ZEA. The dissolution of AFB1 and ZEA in water also facilitates their degradation. At the initial stage of ultrasonic treatment, power ultrasound promotes the dissolution of mycotoxins in water, whereupon they are partially oxidized by free radicals. With a treatment time of 10 min, the reduction rates decreased owing to the dissolution of combined-state mycotoxins. After ultrasonic treatment, the contents of the essential amino acids, the total number of amino acids, and the fatty acids in corn decreased; however, ΔH values decreased during starch gelatinization. In contrast, the amylose content and viscosity of corn significantly increased during gelatinization. Therefore, this method is potentially suitable for the reduction of AFB1 and ZEA contents in corn.