Development of a reusable polymeric fluorescence sensor based on acryloyl ß-cyclodextrin for the determination of aflatoxin B1 in grain products.

AFB1 is a harmful substance that can be found in agricultural products and can seriously affect human health, even in trace amounts. Therefore, monitoring AFB1 levels to ensure food safety and protect public health is crucial. New, highly reliable, selective, and rapid detection methods are needed to achieve this goal. Our work involves the development of a polymeric membrane sensor using radical polymerization that can accurately detect AFB1. Various spectroscopic techniques (Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM)) were used to obtain information about the structural and morphological properties of the prepared sensor. The sensor displayed fluorescence selectively responsive to AFB1 at the excitation wavelength of 376 nm and emission wavelength of 423 nm. The polymeric fluorescence sensor showed good sensitivity and a wide linear range from 9.61 × 10-10 and 9.61 × 10-9 mol/L for AFB1quantification. The limit of detection (LOD) is as low as 3.84 × 10-10 mol/L for AFB1. Other mycotoxins, such as aflatoxin B2 and aflatoxin G1, did not interfere with the sensor's high selectivity towards AFB1. To test the sensor's effectiveness in detecting AFB1 in real samples, three different grain samples - peanuts, hazelnut butter, and peanuts with a sauce known to contain AFB1 - were utilized. The results were satisfactory and demonstrated that the sensor can be successfully employed in real samples, with an error range of 0.43 % to 12.10 %.

Aflatoxin B1 in senna pods from field to storage in Tamil Nadu, India.

Senna, a medicinal herb, is highly susceptible to aflatoxin contamination, which often limits the export value of the final products due to the regulatory limits of importing countries. Presence of aflatoxin B1 (AFB1) was investigated in 229 senna pod samples at various stages from field to storage over three years. Higher incidence of AFB1 was revealed during the late harvest stage with a mean level of 52.4 µg/kg and a range of LOD-206 µg/kg. Out of 48 late harvest samples, 37 exceeded the AFFB1 limit of 2 µg/kg. In contrast, the majority of preharvest samples showed lower levels of AFB1 (1.0 µg/kg). During drying, increase in the mean AFB1 level to 4 µg/kg was observed and it could further increase to 18.1 µg/kg in storage. These results highlight the status of aflatoxin contamination in senna pods and emphasise the importance of implementing good agricultural practices in senna cultivation to mitigate AFB1 contamination.

A multi-verse optimizer-based CNN-BiLSTM pixel-level detection model for peanut aflatoxins.

Peanuts are highly susceptible to contamination by aflatoxins, posing a significant threat to human health. This study aims to enhance the accuracy of pixel-level aflatoxin detection in hyperspectral images using an optimized deep learning method. This study developed a CNN-BiLSTM fusion model optimized by the Multi-Verse Optimizer (MVO) algorithm, specifically designed to detect aflatoxins with high precision. The optimized CNN-BiLSTM model was fine-tuned using aflatoxin spectral data at varying concentrations. The results indicate that the fine-tuned MVO-CNN-BiLSTM model achieved the best performance, with a validation accuracy of 94.92 % and a recall rate of 95.59 %. The accuracy of this model is 6.93 % and 3.6 % higher than machine learning methods such as SVM and AdaBoost, respectively. Additionally, it is 4.18 % and 3.08 % higher than deep learning methods such as CNN and the CNN-LSTM fusion model, respectively. This method enhances pixel-level aflatoxin detection accuracy, supporting the development of online detection devices.

Danshen polysaccharides alleviate AFB1 induced Jejunal injury.

AFB1 is a common foodborne toxin known for its potent carcinogenicity. Danshen polysaccharide (DSP) is an active ingredient of Danshen, which has been demonstrated to possess support intestinal homeostasis and anti-inflammatory activities. We utilized New Zealand White rabbits as an animal model to examine the impact of co-exposure to DSP and AFB1 on the intestines, as well as their underlying mechanisms. The results indicate that DSP elevated the abundance of Oscillospira, Coprococcus, Alistipes, Akkermansia, Bacteroides, Odoribacter, Blautia and Parabacteroides, while decreased the abundance of Sutterella, and Desulfovibrio, correcting AFB1-induced intestinal microbiota dysbiosis and enhancing microbial diversity within the gut. Moreover, DSP reduced the levels of diamine oxidase (DAO), D-Lactate, and malondialdehyde (MDA), while upregulating the expression of total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), zonula occludens-1 (ZO-1), occludin, claudin-4, mucin-2 (MUC2), and secretory immunoglobulin A (sIgA), thereby alleviating the oxidative stress and intestinal barrier dysfunction induced by AFB1. DSP downregulated jejunal lipopolysaccharide (LPS) levels and the mRNA expression and proteins abundance of toll-like receptor 4 (TLR4), myeloiddifferentiationfactor 88 (MyD88), and nuclear factor kappa-B (NF-κB), thereby inhibiting the jejunal inflammation induced by AFB1. In summary, DSP alleviates AFB1-induced jejunal injury by remodeling the gut microbiota, bolstering antioxidant capabilities within the jejunum, fortifying the intestinal barrier, and suppressing the TLR4-mediated release of pro-inflammatory cytokines.

Algal extracts evaluation as an Antitoxicity sustainable solution against aflatoxin B1 toxicity in rat tissues.

Aflatoxin B1 (AFB1) is a pre-carcinogenic molecule produced by toxigenic fungi and is widely harmful to public health. Algae extracts are sub-cellular pilot plants rich in bioactive substances that aid detoxification. This study aimed to reduce AFB1-toxicity in biological tissues of administrated rats using two algae extracts, Spirulina (SPR) and Amphora (AMR). Algae extracts were prepared using an aqueous system, concentrated, and lyophilized before being administrated to rats. The extract contents of total phenolic and flavonoids were determined to indicate their bioactive content and antioxidant potency. The animal experiment was designed in 8 groups as the control negative and control positive (AFB1; 20 µg/kg BW/day); groups 3 and 4 were designed for control positive of algae applied at high doses for toxicity evaluation. Otherwise, four groups were classified as G5 and G6 for rats administrated by AFB1, followed by 50 and 100 mg/kg Spirulina extract, respectively. The G7 and G8 were administrated with an AFB1 dose followed by amphora treatment at 50 and 100 mg extract/kg, respectively. The results showed a significant content of algae extracts of phenolic compounds (27.36 ± 1.75 and 39.55 ± 1.14 mg GAE/g DW for the SPR and AMR, respectively), with a valuable antioxidant activity. For rats treated only with the SPR or AMR extracts, no tissue changes were recorded for the liver, kidney, pancreas, or testis. Again, the biochemical parameters of these groups are recorded without harmful impacts, particularly for the tumor markers of AFP, TNF-α, CEA, and ALP. Once more, a higher extract concentration was more effective in AFB1-toxicity reduction, particularly for the SPR on the liver and kidney tissues. The SPR extract manifested a protective impact in sensitive tissue against the AFB1 effect, particularly in the testis. The results recommend the application of SPR extract at 100 mg/kg bw as an effective treatment for AFB1-toxicity regulation (as pharmaceutical or nutraceutical) involved in daily habits.

Regulation of Conidiation and Aflatoxin B1 Biosynthesis by a Blue Light Sensor LreA in Aspergillus flavus.

Conidia are important for the dispersal of Aspergillus flavus, which usually generates aflatoxin B1 (AFB1) and poses a threat to the safety of agricultural food. The development of conidia is usually susceptible to changes in environmental conditions, such as nutritional status and light. However, how the light signal is involved in the conidiation in A. flavus is still unknown. In this study, LreA was identified to respond to blue light and mediate the promotion of conidiation in A. flavus, which is related to the central development pathway. At the same time, blue light inhibited the biosynthesis of AFB1, which was mediated by LreA and attributed to the transcriptional regulation of aflR and aflS expression. Our findings disclosed the function and mechanism of the blue light sensor LreA in regulating conidiation and AFB1 biosynthesis, which is beneficial for the prevention and control of A. flavus and mycotoxins.

Combining Multiple Omics with Molecular Dynamics Reveals SCP2-Mediated Cytotoxicity Effects of Aflatoxin B1 in SW480 Cells.

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.

Laser-induced CdS/TiO2/graphene dual photoanodes for ratiometric self-powered photoelectrochemical sensor: an innovative approach for aflatoxin B1 detection.

A ratiometric self-powered photoelectrochemical sensor based on laser direct writing technology was constructed to address the problem that the conventional single-signal detection mode was susceptible to the influence of instrumentation and environmental factors, which interfered with the detection results. Laser-induced CdS/TiO2/Graphene was prepared as dual photoanodes (PA1 and PA2), which were controlled by multiplexed switches to form a photocatalytic fuel cell with Pt cathode. By modifying the aptamer of aflatoxin B1 (AFB1) on the photoanode surface, the target was specifically captured to the electrode surface to form a biological complex, which increased the steric hindrance and affected the electron transfer, thus reducing the output signal of the sensor. Targets with different concentrations were incubated on the surface of PA1, and targets with fixed concentrations were incubated on the surface of PA2. Under the control of the multiplex switch, the output signals of the two photoanodes were recorded, and the ratio of these two signals was used as the basis for the quantitative detection of AFB1. The sensor output was linearly increasing with the logarithm of AFB1 concentration from 1.0 to 150 ng mL-1 and the detection limit was 0.0974 ng mL-1. Additionally, this method had good stability, fast response, and good selectivity to real samples, providing an effective method for food safety monitoring.

Impact of Kluyveromyces marxianus VM004 culture conditions on the cell wall structure and its influence on aflatoxin B1 binding.

The aim of this study was to determine the impact of Kluyveromyces marxianus VM004 culture conditions on the cell wall (CW) structure and its influence on aflatoxin B1 binding. The yeast was inoculated into two types of culture media: yeast extract-peptone-dextrose (YPD) broth and dried distiller's grains with solubles (DDG). The CW was extracted from the biomass produced in these media. AFB1 (150ng/ml) adsorption tests using the biomass (1×107cells/ml) and the CW (0.001g) were performed at pH 2 and pH 8. Transmission electron microscopy (TEM) evaluated the CW thickness, and infrared spectroscopy (IR) determined the CW composition. Biomass production in YPD was higher than that in DDG. Cell diameter (µm) and CW thickness (µm) increased in the DDG medium. The CW percentage obtained in DDG was higher than that in YPD. The absorbance of carbohydrates by IR was higher in YPD. pH influenced AFB1 adsorption, which was lower at pH 8. The proportion of ß-glucan and chitin present in CW was higher in the YPD medium. The IR method allowed to study the CW carbohydrate variation under the influence of these carbon sources. In conclusion, the culture media composition influenced the ß-glucan and chitin composition and consequently, AFB1 adsorption.

Biomonitoring and determinants of mycotoxin exposures from pregnancy until post-lactation in HIV-infected and HIV-uninfected women from Harare, Zimbabwe.

The human immunodeficiency virus (HIV) heavily affects women from resource-limited settings who are vulnerable to potentially harmful mycotoxins including aflatoxin B1 (AFB1), fumonisin B1 (FB1) and ochratoxin A (OTA). We aimed to conduct biomonitoring and ascertain the determinants of maternal mycotoxin exposure in pregnancy, lactation and post-lactation periods. We conducted a retrospective longitudinal study in HIV-infected and HIV-uninfected women from Harare, Zimbabwe. 175 and 125 random urine samples in pregnancy and 24 months after delivery (post-lactation) respectively were analysed for aflatoxin M1 (AFM1) and FB1 by ELISA. 6 weeks after delivery (lactation), 226 and 262 breast milk (BM) samples were analysed for AFM1 and OTA respectively by ELISA. The association of demographics and food consumption with mycotoxins was evaluated using multivariable logistic regression. In HIV-infected, urinary AFM1 was detected in 46/94 (Median: 0.05; Range: 0.04-0.46 ng mL-1) in pregnancy and 47/66 (Median: 0.05; Range: 0.04-1.01 ng mL-1) post-lactation. Urinary FB1 was detected in 86/94 (Median: 1.39; Range: 0.17-6.02 ng mL-1) in pregnancy and 56/66 (Median: 0.72; Range: 0.20-3.81 ng mL-1) post-lactation. BM AFM1 was detected in 28/110 (Median: 7.24; Range: 5.96-29.80 pg mL-1) and OTA in 11/129 (Median: 0.20; Range: 0.14-0.65 ng mL-1). In HIV-uninfected, urinary AFM1 was detected in 48/81 (Median: 0.05; Range: 0.04-1.06 ng mL-1) in pregnancy and 41/59 (Median: 0.05; Range: 0.04-0.52 ng mL-1) post-lactation. Urinary FB1 was detected in 74/81 (Median: 1.15; Range: 0.17-6.16 ng mL-1) in pregnancy and 55/59 (Median: 0.96; Range: 0.20-2.82 ng mL-1) post-lactation. BM AFM1 was detected in 38/116 (Median: 7.70; Range: 6.07-31.75 pg mL-1) and OTA in 4/133 (Median: 0.24; Range: 0.18-0.83 ng mL-1). Location, wealth, and peanut butter consumption were determinants of AFB1 exposure. HIV infection, BMI, location, rainy season, unemployment, and age were determinants of FB1 exposure. Women especially those pregnant and/or HIV-infected are at risk of adverse effects of mycotoxins.

Fluorescence assay for aflatoxin B1 based on aptamer-binding triggered DNAzyme activity.

As a kind of mycotoxin, aflatoxin B1 (AFB1), which is often found in agricultural products, poses a threat to human health. Developing a simple sensitive method for AFB1 detection is in great demand. Here, we reported an aptamer-based fluorescence assay for AFB1 detection by using DNAzyme to generate and amplify a signal. We redesigned a pair of DNA sequences, which originated from the anti-AFB1 aptamer and RNA-cleaving DNAzyme 10-23. In the absence of AFB1, the aptamer hybridized with the region of the substrate-binding arm of the DNAzyme, inhibiting the activity of the DNAzyme. In the presence of AFB1, the binding of AFB1 to the aptamer led to the displacement of the DNAzyme from the aptamer. The substrate-binding arm was unblocked, and the activity of the DNAzyme was restored for the hydrolysis of the fluorophore and quencher-labeled substrate, causing a significant fluorescence increase. This assay could detect AFB1 in the dynamic range from 0.98 to 2000 nmol/L with high selectivity, and the detection limit was 0.98 nmol/L. Moreover, the assay was able to detect AFB1 in a complex sample matrix. This work provides a useful tool for the analysis of AFB1.

Tissue inhibitor of metalloproteinase-3 expression affects clinicopathological features and prognosis of aflatoxin B1-related hepatocellular carcinoma.

BACKGROUND: The dysregulation of tissue inhibitor of metalloproteinase-3 (TIMP3) was positively correlated with the progression of hepatocellular carcinoma (HCC). However, it is not clear whether TIMP3 expression is associated with the clinicopathological features and prognosis of aflatoxin B1 (AFB1)-related HCC (AHCC). AIM: To assess the effects of TIMP3 expression on the clinicopathological features and prognosis of AHCC. METHODS: A retrospective study, including 182 patients with AHCC, was conducted to explore the link between TIMP3 expression in cancerous tissues and the clinicopathological characteristics and prognosis of AHCC. TIMP3 expression was detected by immunohistochemistry and its effects on the clinicopathological features and prognosis of AHCC were evaluated by Kaplan-Meier survival analysis and Cox regression survival analysis. Odds ratio, hazard ratio (HR), median overall survival time (MST), median tumor recurrence-free survival time (MRT), and corresponding 95% confidential interval (CI) was calculated to evaluate the potential of TIMP3 expression in predicting AHCC prognosis. RESULTS: Kaplan-Meier survival analysis showed that compared with high TIMP3 expression, low TIMP3 expression in tumor tissues significantly decreased the MST (36.00 mo vs 18.00 mo) and MRT (32.00 mo vs 16 mo) of patients with AHCC. Multivariate Cox regression survival analysis further proved that decreased expression of TIMP3 increased the risk of death (HR = 2.85, 95%CI: 2.04-4.00) and tumor recurrence (HR = 2.26, 95%CI: 1.57-3.26). Furthermore, decreased expression of TIMP3 protein in tissues with AHCC was significantly correlated with tumor clinicopathological features, such as tumor size, tumor grade and stage, tumor microvessel density, and tumor blood invasion. Additionally, TIMP3 protein expression was also negatively associated with amount of AFB1-DNA adducts in tumor tissues. CONCLUSION: These findings indicate that the dysregulation of TIMP3 expression is related to AHCC biological behaviors and affects tumor outcome, suggesting that TIMP3 may act as a prognostic biomarker for AHCC.

Effectiveness of various methods to reduce aflatoxin M1 levels in milk, a systematic review.

The numerous strategies have been conducted worldwide to mitigate the presence of these hazardous toxins. In this systematic study, these researches are summarized. The search of this study was done with keywords aflatoxin M1, AFM1, reduce, decrease, mitigation, prevent, prevention, and milk in databases without a time limit. A total of 49 manuscripts were carefully reviewed, and their data were extracted. Some interventions focused on modifying animal rations, aiming to reduce AFM1 in milk. Some were applied directly to the animals. In this method, which was done more than other research interventions, some toxin binders are used as feed additives. The third type of intervention consisted of measures that were taken directly on the milk itself. Among the three types of interventions, the use of toxin binders in animal feed was more practical and effective.

Reproducible, Accurate, and Sensitive Food Toxin On-Site Detection with Carbon Nanotube Transistor Biosensors.

Field-effect transistor (FET) biosensors based on nanomaterials are promising in the areas of food safety and early disease diagnosis due to their ultrahigh sensitivity and rapid response. However, most academically developed FET biosensors lack real-world reproducibility and comprehensive methodological validation to meet the standards of regulatory bodies. Here, highly uniform and well-packaged semiconducting carbon nanotube (CNT) FET biosensor chips were developed and assessed for the plug-and-play sensing for the rapid and highly sensitive detection of aflatoxin B1 (AFB1) in real food samples to meet international standards. In order to meet the requirements for reproducibility and stability, a scalable residual-free passivation and packaging process was developed for CNT FET biosensors. Portable detection systems were then constructed for on-site detection. The resulting packaged chips were functionalized with nucleic aptamers to enable highly selective detection of AFB1 in food samples with a detection limit (LOD) of 0.55 fg/mL (standard) for AFB1 and cross-reactivity coefficients to interferences as low as 1.8 × 10-7 in simulated solutions. Utilizing the portable detection system, on-site real food detection was achieved with a rapid response time less than 60 s, and LOD of 0.25 pg/kg (standard) in complex corn sample matrices. Single-blind tests demonstrated the ability of the chips to detect AFB1-positive food with 100% accuracy, using a set of 30 peanut samples. Validation experiments confirmed that the detection range, stability, and repeatability met international standards. This study showcased the accuracy, reliability, and potential practical applications of CNT FET biosensor chips in areas such as food safety and rapid biomedical testing.

Contamination of the traditional medicine Radix Dipsaci with aflatoxin B1 impairs hippocampal neurogenesis and cognitive function in a mouse model of osteoporosis.

BACKGROUND: Aflatoxin B1, which can penetrate the blood-brain barrier and kill neural cells, can contaminate traditional herbal medicines, posing a significant risk to human health. The present study examined cellular, cognitive and behavioral consequences of aflatoxin B1 contamination of the anti-osteoporotic medicine Radix Dipsaci. METHODS: A mouse model of osteoporosis was created by treating the animals with all-trans-retinoic acid. Then the animals were treated intragastically with water decoctions of Radix Dipsaci that contained detectable aflatoxin B1 or not. The animals were compared in terms of mineral density and mineral salt content of bone, production of pro-inflammatory factors, neurogenesis and microglial activation in hippocampus, as well as behavior and cognitive function. RESULTS: Contamination of Radix Dipsaci with aflatoxin B1 significantly reduced the medicine's content of bioactive saponins. It destroyed the ability of the herbal decoction to improve mineral density and mineral salt content in the bones of diseased mice, and it induced the production of the oxidative stress marker malondialdehyde as well as the pro-inflammatory cytokines interleukin-1ß and tumor necrosis factor-α. Aflatoxin B1 contamination inhibited formation of new neurons and increased the proportion of activated microglia in the hippocampus. These neurological changes were associated with anhedonia, behavioral despair, and deficits in short-term memory and social memory. CONCLUSION: Contamination of Radix Dipsaci with aflatoxin B1 not only eliminates the herbal decoction's anti-osteoporotic effects, but it also induces neurotoxicity that can lead to cognitive decline and behavioral abnormalities. Such contamination should be avoided through tightly regulated production and quality control of medicinal herbs.

Hesperetin alleviates aflatoxin B1 induced liver toxicity in mice: Modulating lipid peroxidation and ferritin autophagy.

One of the ways Aflatoxin B1 damages the liver is through ferroptosis. Ferroptosis is characterized by the build-up of lipid peroxides and reactive oxygen species (ROS) due to an excess of iron. Dietary supplements have emerged as a promising strategy for treating ferroptosis in the liver. The flavonoid component hesperetin, which is mostly present in citrus fruits, has a number of pharmacological actions, such as those against liver fibrosis, cancer, and hyperglycemia. However, hesperetin's effects and mechanisms against hepatic ferroptosis are still unknown. In this study, 24 male C57BL/6 J mice were randomly assigned to CON, AFB1 (0.45 mg/kg/day), and AFB1+ hesperetin treatment groups (40 mg/kg/day). The results showed that hesperetin improved the structural damage of the mouse liver, down-regulated inflammatory factors (Cxcl1, Cxcl2, CD80, and F4/80), and alleviated liver fibrosis induced by aflatoxin B1. Hesperetin reduced hepatic lipid peroxidation induced by iron accumulation by up-regulating the levels of antioxidant enzymes (GPX4, GSH-Px, CAT, and T-AOC). It is worth noting that hesperetin not only improved lipid peroxidation but also maintained the dynamic balance of iron ions by reducing ferritin autophagy. Mechanistically, hesperetin's ability to regulate ferritin autophagy mostly depends on the PI3K/AKT/mTOR/ULK1 pathway. In AFB1-induced HepG2 cells, the addition of PI3K inhibitor (LY294002) and AKT inhibitor (Miransertib) confirmed that hesperetin regulated the PI3K/AKT/mTOR/ULK1 pathway to inhibit ferritin autophagy and reduced the degradation of ferritin in lysosomes. In summary, our results suggest that hesperetin not only regulates the antioxidant system but also inhibits AFB1-induced ferritin hyperautophagy, thereby reducing the accumulation of iron ions to mitigate lipid peroxidation. This work provides a fresh perspective on the mechanism behind hesperetin and AFB1-induced liver damage in mice.

Isolation of Bacillus licheniformis and its protective effect on liver oxidative stress and apoptosis induced by aflatoxin B1.

Aflatoxin B1 (AFB1) is one of the most toxic mycotoxins. The use of probiotics is an effective approach to reduce aflatoxins content in foods. To find efficient bacterial species that can eliminate or detoxify AFB1, a bacterial strain S51 capable of degrading AFB1 was isolated from chicken intestine and soil samples by using a culture medium containing coumarin as the sole carbon source. Based on the results of 16S rRNA gene sequence analysis, this isolate (strain S51) was identified as Bacillus licheniformis strain QT338. Further characterization of strain S51 showed that it could degrade AFB1 by 61.3% after incubation at 30°C for 72 h. Additional studies demonstrated that S51 promoted good growth performance of the treated chickens, showed no hemolytic activity, carried few drug resistance genes, and exhibited a certain level of tolerance to acid and bile salts. Furthermore, to verify whether strain S51 exerts a protective effect on AFB1-induced liver injury in chickens and to elucidate the underlying mechanism, a chicken toxicity model was induced with AFB1 (100 µg/kg BW) and treated with S51(1×109CFU/mL) for 12 d. The results showed that S51 decreased the level of alanine transaminase, aspartate transaminase, and total bilirubin (P < 0.05); increased glutathione activity and total antioxidant capacityin the liver induced by AFB1, and decreased malondialdehyde production (P < 0.05). S51 also up-regulated the mRNA expression level of the antioxidant proteins HO-1 and Nrf2 and down-regulated the expression of the oxidation-related factor Keap1 in the Nrf2/Keap1 signaling pathway (P <0.05). S51 inhibited hepatocyte apoptosis induced by AFB1 and decreased the mRNA expression levels of the apoptosis-related genes Bax, caspase-3, caspase-9, and Cyt-C (P < 0.05). These results indicate that S51 regulates apoptosis and alleviates AFB1-induced oxidative stress in chicken liver by controlling the Nrf2/Keap1 signaling pathway.

Licochalcone A mitigates aflatoxin B1-induced immunotoxicity via ferroptosis in bursa of broilers and macrophages.

Aflatoxin B1 (AFB1) is a mycotoxin which is responsible for severe damage to the immune system of humans and livestock. Licochalcone A (Lico A), a polyphenol derived from turmeric, has attracted great attention due to its wonderful antioxidant properties. Ferroptosis, an iron-dependent cell death related to oxidative stress, which plays a crucial role in the resistance of phytochemical to immune-associated injury. Nevertheless, effects of Lico A on the bursa of broilers exposed to AFB1 remain unclear. In this work, broilers were fed diets supplemented with 2 mg/kg of AFB1 and 50 mg/kg of Lico A. Meanwhile, various concentrations of Lico A and AFB1 (15 µM) were used to stimulate macrophages. These results revealed that AFB1 resulted in more severe bursa atrophy and relative weight reduction; the expression of pro-ferroptosis protein ACSL4 and the content of malondialdehyde (MDA) were significantly elevated, while the expression of anti-ferroptosis proteins GPX4, xCT, FSP1 and the content of Glutathione (GSH) was obviously reduced. However, Lico A treatment effectively reversed these effects in the bursa of broilers. Meanwhile, in bursa and macrophages, Lico A mitigated the expression of AFB1-induced apoptosis-associated protein (Caspase-3, Bax, Bcl-2) as well as antioxidant protein (Nrf2, GCLM, HO-1). Importantly, ferroptosis was also observed in macrophages induced by AFB1. Lico A efficaciously alleviated AFB1-induced mitochondrial membrane potential decrease and reactive oxygen species (ROS) production in macrophages; in contrast, Lico A evidently inhibited AFB1-triggered ROS generation and cytotoxicity, which was disabled by the addition of Erastin. Moreover, Liproxstatin-1 significantly inhibited ROS generation induced by AFB1. In summary, the present study elucidates that the main mechanism by which Lico A attenuates AFB1-induced immunotoxicity is through the suppression of ferroptosis, apoptosis, mitochondrial damage and oxidative stress, which is promising for the improvement of immunotoxic effects of AFB1.

When less is more: The association between the expression of polymorphic CYPs and AFB1-induced HCC.

BACKGROUND: An individual's genetic fingerprint is emerging as a pivotal predictor of numerous disease- and treatment-related factors. Single nucleotide polymorphisms (SNPs) in drug-metabolizing enzymes play key roles in an individual's exposure to a malignancy-associated risk, such as Aflatoxin B1 (AFB1)-induced hepatocellular carcinoma (HCC). AIM: This study aimed at reviewing literature on the polymorphisms that exist in CYP enzymes and their possible link with susceptibility to AFB1-induced HCC. MATERIALS & METHODS: A set of keywords associated with the study subject of interest was used to search the Google Scholar and the PubMed database. The last ten years' worth of research projects were included in the results filter. The research involved HCC patients and any connection between polymorphic forms of CYP enzymes and their susceptibility to AFB1-induced HCC, including older but significant data. RESULTS: Variations in CYP1A2 and CYP3A4 were reported to impact the rate and magnitude of AFB1 bio-activation, thus influencing an individual's vulnerability to develop HCC. In HCC patients, the activity of CYP isoforms varies, where increased activity has been reported with CYP2C9, CYP2D6, and CYP2E1, while CYP1A2, CYP2C8, and CYP2C19 exhibit decreased activity. CYP2D6*10 frequency has been discovered to differ considerably in HCC patients. Rs2740574 (an upstream polymorphism in CYP3A4 as detected in CYP3A4*1B) and rs776746 (which affects CYP3A5 RNA splicing), both of which influence CYP3A expression, thus impacting the variability of AFB1-epoxide adducts in HCC patients. DISCUSSION: CYP1A2 is the primary enzyme accountable for the formation of harmful AFBO globally. CYP3A4, CYP3A5, CYP3A7, CYP2B7, and CYP3A3 are also implicated in the bio-activation of AFB1 to mutagenic metabolites. It is thought that CYP3A4 is the protein that interacts with AFB1 metabolism the most. CONCLUSION: Polymorphic variants of CYP enzymes have a functional impact on the susceptibility to AFB1-induced HCC. Outlining such variation and their implications may provide deeper insights into approaching HCC in a more personalized manner for guiding future risk-assessment, diagnosis, and treatment.

Factors influencing aflatoxin B1 levels in the groundnut (Arachis hypogaea L.) germplasm of Ethiopia.

As there was no maximum permissible limit prescription for aflatoxin B1 (AFB1) in Ethiopia, this study has been conducted to generate data on AFB1 levels in Ethiopian groundnut accessions/landraces. Besides, an attempt was made to find out if there is any relationship between AFB1 and other parameters such as altitude of cultivation, individual seed weight, kernel colonization by Aspergillus flavus, total carbohydrates, protein and total free amino acids. Out of the 28 accessions studied, merely six accessions registered ≤2 ppb AFB1 and thus, they comply with maximum permissible limit set by European Union. Altitude of cultivation had no relationship with AFB1 levels. Interestingly, total carbohydrates in the seeds as well as kernel colonization by A. flavus showed statistically significant (p < 0.01) positive relationships with AFB1 levels. It is suggested to use kernel colonization measurement as an alternative to the expensive ELISA based AFB1 measurement. Besides, suitable pre- and post-harvest aflatoxin management strategies should be developed to alleviate the AFB1 levels in Ethiopian groundnut.