Aflatoxin B1 Research Articles

Exposure of pregnant and lactating parental mice to aflatoxin B1 promotes hepatotoxicity in offspring mice.

Aflatoxin B1 (AFB1) taints feeds stuffs, endangering livestock's health and resulting in the liver and breast damage. At the same time, while breastfeeding, AFB1 crosses the mammary glands and enters the milk, harming the offspring. This study investigated the liver damaging effects of maternal AFB1 exposure during pregnancy and lactation in offspring mice. The livers of 8-day-old offspring mice were obtained from female mice who were administered AFB1 (2mg/kg) 1week prior to and 1week following birth. The results showed that AFB1 increased the levels of malondialdehyde (MDA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), pro-inflammatory-related proteins (iNOS, COX-2, IL-6), and apoptosis-related proteins (Caspase-3, Caspase-9, Bax) by AFB1-induced in liver of offspring mice. Furthermore, the use of F40/80, HE, and TUNEL staining further demonstrated the existence of inflammation and apoptosis in the liver. Intriguingly, in the liver of offspring mice, AFB1 increased antioxidant protein and inhibit ferroptosis-related protein activity (FTH, GPX4), mitochondrial function-associated proteins (UQCRC2, COX IV, Cyt C), lipid metabolism-associated proteins (HMGCR, SPEBE1, FAS), and autophagy-related proteins (Atg7, Beclin-1, LC3I/II) in the liver of mice. In conclusion, AFB1 enters the liver of offspring mice through milk, which in turn causes liver injury. This outcome explains how AFB1 exposure affects female animals and their progeny and lays the strategy for livestock prevention.

Degradation of aflatoxin B1 in corn by water-assisted microwave irradiation and its kinetic.

Aflatoxin B1 (AFB1) is a highly toxic, carcinogenic, teratogenic, and mutagenic mycotoxin commonly found in corn. In this work, water-assisted microwave irradiation (WMI1) was used to degrade AFB1 in corn, during which the influencing factors and kinetics of AFB1 degradation were also studied. The results showed that the degree of corn crushing, the heating rate of WMI, the temperature of WMI, the solid-liquid ratio, the initial content of AFB1 and the microwave power were all important factors affecting the degradation of AFB1. With the increase of WMI temperature, AFB1 content in corn, microwave power and the decrease of solid-liquid ratio, the degradation rate of AFB1 in corn by WMI rose continuously. Its maximum degradation rate was more than 90.6%. Meanwhile, the degradation kinetics of AFB1 in corn revealed that the degradation process of WMI followed a pseudo-first-order kinetic equation. It was demonstrated that water molecules not only acted as solvents for AFB1 but also actively participated in its degradation process within corn samples during WMI treatment. Consequently, the results indicated that WMI was an effective method for degrading AFB1 in corn.

The impact of aflatoxin B1 on animal health: Metabolic processes, detection methods, and preventive measures.

Aflatoxin (AF) is a toxic metabolite produced by the fungus Aspergillus. The various subtypes of AFs include B1, B2, G1, G2, M1, and M2, with Aflatoxin B1 (AFB1) being the most toxic. These AFs are widespread in the environment, particularly in soil and food crops. The World Health Organization (WHO) has classified AFB1 as a highly potent natural Class 1A carcinogen. Excessive exposure to AFB1 can lead to poisoning in both humans and animals, posing substantial risks to food safety and livestock breeding industries. This review provides an overview of the metabolic processes, detection methods, and the detrimental impacts of AFB1 on animal reproduction, immunity, nerves, intestines, and metabolism. Furthermore, it explores the preventive and control capacities of natural active substances, trace elements, and microorganisms against AFB1. Ultimately, this paper serves as a reference for further research on the pathogenic mechanism of AFB1, the development of preventive drugs, and the selection of effective detoxification measures for AFB1 in animal feed.

Applying hollow octahedron PtNPs/Pd-Cu2O nanozyme and highly conductive AuPtNPs/Ni-Co NCs to colorimetric -electrochemical dual-mode aptasensor for AFB1 detection.

Aflatoxin B1 (AFB1) is a secondary metabolite produced by Aspergillus flavus and Aspergillus parasiticus. This toxin is highly carcinogenic and toxic, posing a serious threat to human and animal health. AFB1 primarily enters the human body through contaminated food, particularly peanuts, corn, nuts, and wheat. These foods are easily contaminated with AFB1 during planting, harvesting, storage, and processing. Therefore, the establishment of an accurate and sensitive method for the detection of AFB1 is essential to safeguard food safety and human health. This study applied catalytic hairpin assembly (CHA), hybridization chain reaction (HCR), hollow octahedron PtNPs/Pd-Cu2O bifunctional nanozyme, and AuPtNPs/Ni-Co NCs to develop a colorimetric-electrochemical dual-mode aptasensor for AFB1 detection. Here, Pd-Cu2O corner-etched octahedra with cavities and oxygen vacancy defects were first designed, followed by the synthesis of PtNPs/Pd-Cu2O through the loading of PtNPs to enhance the peroxidase-like activity of Pd-Cu2O. PtNPs/Pd-Cu2O effectively catalyzed the generation of blue oxidation products from 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide (H2O2). It also enhances the redox reaction of methylene blue (MB) at the electrode and improves the electrochemical signal. AuPtNPs/Ni-Co NCs possess a large specific surface area and excellent conductivity, enhanced the redox signal of MB. Additionally, the aptasensor combined the enzyme-free amplification strategies of CHA and HCR to improve sensitivity. The aptasensor reached limit of detection (LOD) 0.76pg/mL in electrochemical mode and 0.49pg/mL in colorimetric mode. The designed colorimetric-electrochemical dual-mode aptasensor exhibits high selectivity, specificity, stability, and reliability. The dual-mode aptasensor had been successfully applied to the spiked analysis of edible oils, peanuts, and cornmeal, demonstrating excellent recovery rates. Therefore, the dual-mode aptasensor had a wide application prospect in the field of quality supervision and food safety.

Role of red beetroot in bread for reducing mycotoxin risks: Bioavailability of beetroot polyphenols and betalains with ochratoxin a, aflatoxin B1 and zearalenone in Caco-2 cells.

The interaction between dietary bioactive compounds and mycotoxins in food safety is crucial due to the potential health risks raised by mycotoxins and the protective functions of bioactive substances. This study is focused on red beetroot (Beta vulgaris), a rich source of polyphenols and betalains, incorporated into a daily consumption food such as bread, to examine its effects on the bioavailability of mycotoxins using an in vitro Caco-2 cell model. This study investigates how these compounds affect the bioavailability of mycotoxins, specifically ochratoxin A (OTA), aflatoxin B1 (AFB1), and zearalenone (ZEA), which are known to compromise intestinal barrier function and nutrient absorption. Additionally, bioaccesibility and bioavailability of total betalains (betacyanins and betaxanthins) (TBC) and polyphenols (TPC) content was evaluated. The beetroot-enriched breads were subjected to an in vitro digestion process, followed by a transepithelial transport assay to assess the bioavailability in differentiated Caco-2 cells. Results indicate an increase in the bioaccesibility of TBC and TPC (up to 99% and 27%, respectively) during digestion, suggesting enhanced absorption and protective effects against mycotoxin-induced damage. The presence of beetroot bread polyphenols and betalains increased the bioavailability of mycotoxins, with complex interactions observed, particularly in triple mycotoxin's combination. The results highlight the complex interactions between dietary components and mycotoxins' bioavailability, underlining the importance of further research into their mechanisms of action and potential applications in food safety and nutrition.

Comprehensive review for aflatoxin detoxification with special attention to cold plasma treatment.

Aflatoxins are potent carcinogens and pose significant risks to food safety and public health worldwide. Aflatoxins include Aflatoxin B1 (AFB1), Aflatoxin B2 (AFB2), Aflatoxin G1 (AFG1), Aflatoxin G2 (AFG2), and Aflatoxin M1 (AFM1). AFB1 is particularly notorious for its carcinogenicity, classified as a Group 1 human carcinogen by the International Agency for Research on Cancer (IARC). Chronic exposure to aflatoxins through contaminated food and feed can lead to liver cancer, immunosuppression, growth impairment, and other systemic health issues. Efforts to mitigate aflatoxin contamination have traditionally relied on chemical treatments, physical separation methods, and biological degradation. However, these approaches often pose challenges related to safety, efficacy, and impact on food quality. Recently, cold plasma treatment has emerged as a promising alternative. Cold plasma generates reactive oxygen species, which effectively degrade aflatoxins on food surfaces without compromising nutritional integrity or safety. This reviewconsolidatescurrent research and advancements in aflatoxin detoxification, highlighting the potential of cold plasma technology to revolutionize food safety practices. By exploring the mechanisms of aflatoxin toxicity, evaluating existing detoxification methods, and discussing the principles and applications of cold plasma treatment.

A series of colorimetric and self-calibrated fluorescent sensors based on Ln-BTCs for detection of aflatoxin B1 in food

A series of colorimetric and self-calibrated fluorescent sensors based on Ln-BTCs for detection of aflatoxin B1 in food

A dual-mode aptasensor for aflatoxin B1 determination based on photoelectrochemistry and upconversion fluorescence resonance energy transfer

A dual-mode aptasensor for aflatoxin B1 determination based on photoelectrochemistry and upconversion fluorescence resonance energy transfer

Degradation of aflatoxin B1 by X-ray irradiation

Degradation of aflatoxin B1 by X-ray irradiation

Magnetically controlled fluorescence biosensor for simultaneous detection of aflatoxin B1 and its toxin-producing gene aflD

Magnetically controlled fluorescence biosensor for simultaneous detection of aflatoxin B1 and its toxin-producing gene aflD

Efficacy of advanced chelate technology-based 7-mineral supplementation in mitigating aflatoxin B1-induced impairments in broiler chicken performance and intestinal health.

Optimal levels and bioavailability of trace minerals (TM) in the broiler diet are important for improving performance and health status in the presence of dietary toxins. The study examines the effectiveness of advanced chelate technology-based 7-minerals (ACTM) in broilers fed aflatoxin B1 (AFB1)-contaminated diets, involving 768 chickens in eight treatments with six replicates, following a completely randomized design. Treatments contained (1) negative control (NC) group receiving a basal diet without AFB1 and containing recommended inorganic TM (ITM) levels (NC+ITM), (2) positive control (PC) group receiving a basal diet with 0.5mg AFB1/kg and recommended ITM levels (PC+ITM), (3) PC diet+toxin binder (ITM+TB), (4 and 5) PC diet with 50% and 100% ACTM instead of ITM (ACTM50 and ACTM100), (6 and 7) PC diet with 12.5% and 25% extra ACTM (ITM+ACTM12.5 and ITM+ACTM25), and (8) PC diet with 125% ITM levels (ITM125). The results showed that the ACTM100 and ITM+ACTM25 treatments resulted in higher average weight gain and European production efficiency index compared to the PC+ITM treatment, but lower than the NC+ITM treatment. Key indicators of gut health, such as ileal digestibility of crude fat and phosphorus, AMEn value, duodenal villus height to crypt depth ratio, villus surface area, and gene expression of junctional adhesion molecule 2 were significantly improved in the ACTM100, ITM+BT, and NC+ITM groups compared to the PC+ITM group. Additionally, jejunal occludin expression increased in the ACTM100, ITM+ACTM25, ITM+TB treatments, and the jejunal zonula occludens-1 expression increased significantly in the ACTM100 and ITM+ACTM25 groups. The results indicate that completely replacing ITM with ACTM or adding ACTM supplement to ITM diets at 25% extra commercial levels can improve growth performance, gut health, and nutrient digestibility in the presence of AFB1 challenge. These effects are comparable to diets containing a commercial toxin binder.

Probiotic-based adsorbent mitigates aflatoxin B1 toxicity in Piaractus mesopotamicus: assessing well-being via changes in tissue architecture and digestive enzyme activity.

Aflatoxin is a mycotoxin produced by fungi of the genus Aspergillus that is present in various foods. Probiotics are well-established products in aquaculture, and due to their effective contribution to the intestine, they can be used as an aflatoxin adsorbent. This study evaluated the effects of aflatoxin B1 (AFB1) on enzymatic activity and intestinal function in Piaractus mesopotamicus (pacu) fingerlings fed diets containing a probiotic-based adsorbent (PBA). Seventy-two fish with an average weight of 12 ± 1.30g were used in the study. The experiment was conducted over 15 days using a completely randomized design with six diet treatments (AFB1 per kg of formulated diet) and two replicates. These treatments were: control without AFB1; 25.0µg of AFB1; 400.0µg of AFB1; control diet without AFB1 + PBA; 25.0µg of AFB1 + PBA; and 400.0µg of AFB1 + PBA. After the experimental period, the digestive enzymes protease, amylase, and lipase from the stomach, pyloric caeca, and intestine were quantified. The height and width of the intestinal villi, pyloric caeca, and stomach wall were measured. Fish fed the 400.0µg of AFB1 diet showed reduced feed consumption, even though they ingested higher amounts of AFB1 compared to those fed the other experimental diets. The best zootechnical performance parameters were observed in fish fed the control diet without AFB1 + PBA. Changes (p < 0.05) were observed in the amount of protease in the stomach, pyloric caeca, and intestine; in the amount of amylase in the intestine; and the amount of lipase in the pyloric caeca and intestine. Changes were observed (p < 0.05) in the length of the stomach wall and the anterior and posterior intestines. Changes in the width of the stomach and anterior intestine walls were observed as a result of diet. Our results suggest that the use of probiotics as an aflatoxin adsorbent in pacu diets is beneficial from a physiological perspective and may also enhance growth.

Vitamin B6 Alleviates Aflatoxin B1-Induced Impairment of Testis Development by Activating the PI3K/Akt Signaling Pathway.

Aflatoxin B1 (AFB1) is a harmful environmental contaminant known to disrupt gut microbiota and cause health problems. In recent years, the role of vitamin B6 (VB6) in maintaining intestinal and reproductive health has attracted much attention. AFB1 has been found to damage the intestinal barrier and cause inflammation by disrupting the intestinal microbiota, particularly by increasing the abundance of Enterococcus. In mice treated with AFB1, serum metabolites were disturbed, VB6 serum levels were reduced, and testicular inflammation was exacerbated. Enterococcus exposure in mice leads to a reduction in serum VB6 levels, which is accompanied by intestinal and testicular damage. However, VB6 supplementation significantly ameliorated AFB1-induced intestinal and testicular injury. Transcriptomics and Western blotting showed that after AFB1 exposure, VB6 could increase the expression of phosphoinositide 3-kinase (PI3K) and kinase B (AKT) as well as their phosphorylated forms in the testes of mice. Based on the results, AFB1 leads to intestinal and testicular damage by disturbing the gut microbiota, and VB6 represents a potential therapeutic to counteract this damage. In conclusion, supplementation with adequate VB6 can ameliorate AFB1-induced intestinal and testicular damage, emphasizing the importance of VB6 intervention and providing a new perspective for the prevention and treatment of AFB1-related health issues.

Quality of maize supplied under school feeding program and associated financial losses in Turkana County, Kenya

Turkana County, being an arid and semi-arid county is one of the major beneficiaries of the homegrown school feeding program from the government of Kenya. Most of the maize consumed in Turkana county is sourced from other counties as well as neighbouring countries such as Uganda and Ethiopia. Due to the vast distance, high temperatures as well as security challenges, post-harvest losses in the county are inevitable. This study, therefore, sought to establish the losses associated with the quality and safety of maize supplied to public primary schools under the homegrown school feeding program. All suppliers, n=22, who were supplying maize to public primary schools under the homegrown school feeding program, n=128 were included in the study. Maize samples were picked from school stores and analyzed for moisture content, aflatoxin B1, total aflatoxin and fumonisin. Moisture content was determined by AOAC method number AOAC 976.08:2012 while aflatoxin and fumonisin were determined using the ELISA method. Losses were estimated based on the Kenyan standard requirements for maize where any result above the recommended limit was considered a loss. The market price of maize at the time of study, Ksh. 50 per Kg was used to estimate the associated financial losses. The quality losses were Ksh. 15,075,950/= for moisture content and Ksh. 25,805,350/= for grading and live infestation. Safety losses based on aflatoxin B, Total aflatoxin and fumonisin were Ksh. 6,726,850/=, Ksh. 4,362,500/= and Ksh. 1,356,050/=, respectively. In total, Ksh. 53 million would be lost by the county government if the Kenyan standard for maize was well enforced. This shows that the quality and safety of maize supplied to schools under the school meals program is questionable and contributes further to post-harvest losses. Therefore, there is a need for proper sensitization on post-harvest handling among the school suppliers as well as investment in testing infrastructure by the county government. Key words: Feeding, Losses, school, safety, quality, post-harvest, maize, financial, supplied

ОЦЕНКА ОБЪЕКТИВНЫХ МОРФОЛОГИЧЕСКИХ ПРИЗНАКОВ МИТОХОНДРИЙ ГЕПАТОЦИТОВ КРЫС ПРИ СОЧЕТАННОМ МИКОТОКСИКОЗЕ НА ФОНЕ ПРИМЕНЕНИЯ ПРОФИЛАКТИЧЕСКИХ КОМПЛЕКСОВ

The purpose of research is to study the ultrastructure of the liver and morphometric characteristics of mitochondria of white rats with combined mycotoxicosis against the background of the use of prophylactic complexes. The work was performed on 80 rats weighing 150–170 g, divided into 8 groups (n=10). The first group of rats served as biological control. Mycotoxins were given to rats with food (aflatoxin B1 – 2.5 mg/kg; T-2 toxin – 5 mg/kg; zearalenone – 2.0 mg/kg of feed). In addition to the toxic diet, rats of groups 3–5 received prophylactic complexes at a dose of 0.25 % of the diet (prophylactic complexes No. 1, 2, 3: 1 – β-glucans, milk thistle meal, vitamin E, ascorbic acid, levamisole; 2 – bentonite, succinic acid, methyluracil; vitamin A, probiotic preparation Florin; 3 – halloysite, methionine, β-glucans, milk thistle meal). Rats of groups 6–8, in addition to the main diet, received prophylactic complex № 1, 2, and 3, respectively, at a dosage similar to that for groups 3–5. Ultrastructural studies of the liver were performed according to standard electron microscopic techniques. Combined mycotoxicosis (group No. 2) at the level of the ultrastructure of hepatocytes is manifested by an uncharacteristic shape of nuclei, redistribution of chromatin, destructive processes in mitochondria, and clearing of significant areas of the cytoplasm. The electron-light cytoplasm has few inclusions and a fragmented endoplasmic reticulum. The matrix of most mitochondria has an average electron density, cristae are not visualized, some mitochondria are characterized by membrane destruction. In a comparative aspect, the toxic control group compared to the biological group is characterized by a statistically significant (in the Mann-Whitney test with Bonferroni correction p&lt;0.0096) decrease in all values of the morphometric characteristics of mitochondria (area, perimeter, sphericity and caliperometric diameter). Approbation of prophylactic complexes in case of combined mycotoxicosis showed a protective and therapeutic effect (presence of glycogen, sufficiently dense cytoplasm, preservation of the integrity of membrane organelles).

Mycotoxin binder for immune and intestinal histopathology ameliorations against Newcastle disease in vaccinated broilers

Background In broiler farming, vaccination against Newcastle disease (ND) is essential. Nonetheless, during the post-vaccination phase, production may be negatively impacted by mycotoxin contamination in feed. This study aimed to evaluate the effect of mycotoxin binders on immune and intestinal histopathology ameliorations against newcastle disease in vaccinated broilers with aflatoxin B1 (AFB1) and ochratoxin A (OTA) toxication. Methods A total of 20 broilers were randomly assigned into 4 groups with 5 replications i.e. (C-) broiler groups with basal feed, (C+) broiler groups with AFB1 and OTA feed contamination, (T1) and (T2) broiler groups with exposed AFB1, OTA, and toxin binders as feed additives with dose 1.1 g/kg and 1.6 g/kg feed, respectively. ND vaccination was carried out on day 7 and 21. Antibody titers were evaluated from serum samples of broiler on days 14, 28, and 35 for further hemagglutination inhibition (HI) test. Histopathology of the cecum and colon organs was evaluated using HE staining on day 36. HI test and histological scoring were analyzed using the One-Way ANOVA, followed by Duncan’s test with a p &lt; 0.05 in SPSS v.26 software (IBM Corp., Armonk, NY). Results As a result, histopathological improvement of the cecum and colon was reported based on mucosal rupture, hemorrhage and necrosis on day 35. An increase in the mean antibody titer compared to days 14 and 28 was observed on day 35, with significant changes observed in serum samples based on the C+ group, which was significantly different from the C- and T2 groups (p &lt; 0.05). Conclusions This study revealed that a 1.6 g/kg toxin binder dose in feed can increase antibody titer and histopathology of cecum and colon in broiler chickens after ND vaccination fed with mycotoxin-contaminated feed.

Synbiotics effects of d-tagatose and Lactobacillus rhamnosus GG on the inflammation and oxidative stress reaction of Gallus gallus based on the genus of cecal bacteria and their metabolites.

Abuse of feed supplement can cause oxidative stress and inflammatory responses in Gallus gallus. Synbiotics are composed of prebiotics and probiotics and it possess huge application potentials in the treatment of animal diseases. This study examined the effect of d-tagatose on the probiotic properties of L. rhamnosus GG, L. paracasei, and S. lactis so as to screen the best synbiotic combinations. Treat Gallus gallus exhibiting oxidative stress and immune response caused by aflatoxin b1 with optimal synbiotics for 14 days, detect the changes of inflammatory markers and oxidative stress markers of Gallus gallus using qRT-PCR, and identified the intestinal bacteria genera and their metabolites in the cecum of Gallus gallus using gut microbiota and metabolomics analysis. The results indicated that oxidative stress and immune response factor expressions quantity in Gallus gallus decreased significantly after 14 days of treatment, compared with model group, the low-dose treatment group's SOD1, SOD3, GPX1, GPX2, GSR, H6DP, and HO-1 genes in liver were downregulated by 36.03%, 40.01%, 45.86%, 40.79%, 37.68%, 25.04%, and 29.89%, the IL-1, IL-2, IL-4, IL-6, IgA, IgM, and IgG genes in blood and spleen were downregulated by 26.59%, 34.19%, 21.19%, 28.18%, 35.93%, 12.67%, 21.81 and 35.93%, 22.85%, 21.19%, 28.78%, 35.93%, 15.36%, 29.73%. The intestinal bacteria genera and metabolomics analysis results indicated that the abundance of beneficial bacteria genus was up-regulated, and the proportion of pathogenic bacteria genera decreased. The amount of beneficial metabolites associated with antioxidant and anti-inflammatory effects was upregulated. The synbiotic composed of d-tagatose and L. rhamnosus GG can treat oxidative stress and immune response by altering the structure of intestinal bacteria genera and the production of metabolites.

Effects of Green-Synthesised Copper Oxide–Zinc Oxide Hybrid Nanoparticles on Antifungal Activity and Phytotoxicity of Aflatoxin B1 in Maize (Zea mays L.) Seed Germination

Maize contamination with aflatoxin B1 (AFB1) is of significance on a global scale due to its major contribution to food security. It is very probable that substantial amounts of AFB1 may be absorbed by germinating seeds grown in contaminated soil and cause deleterious effects on the growth and development of maize. In this study, the effect of green-synthesised ZnO-CuO hybrid nanoparticles (NPs) on antifungal activity and reducing the toxic effects of AFB1 on seed germination was examined. A notable inhibitory effect of green-synthesised ZnO-CuO nanoparticles (NPs) on A. flavus was observed at a concentration of 0.5 ppm, resulting in 13.1% inhibition, which was more effective than the higher concentration of 1.0 ppm and the control. The results showed that the final germination percentage of the seeds that were inoculated with 320 ppb was significantly increased by the treatment with 125 mg/mL of green ZnO-CuO hybrid NPs. This study indicated the potential of green-synthesised ZnO-CuO hybrid NPs as alternative antifungal agents to control aflatoxin production in maize to improve food security and safety by supressing the threat posed by AFB1.

Phylogenetic and Gene Expression of Norsolorinic Acid Analysis of Aflatoxigenic Strains and Quantification of Aflatoxin B1 Levels into White Rice Grains in Kenya

The most important pathogens of global significance in food security are mycotoxigenic fungi. Aflatoxigenic strains contamination in rice grains can lead to a health risk for consumers. Under appropriate conditions, aflatoxigenic fungi can grow and subsequently produce aflatoxins. Aflatoxin poses a real threat to food safety and security and destroys approximately 25 % of the world's food crops and every year 420,000 people die from consuming contaminated food causing an estimated loss of 33 million healthy life years. This study screened Norsoloric Acid gene involved in the biosynthesis pathway, gene expression profile of aflatoxigenic strains characterized from rice grains, and assessed aflatoxin B1 levels in white rice grains. We combine the use of molecular techniques and Enzyme-Linked Immunosorbent Assay to detect aflatoxin biosynthesis aflD gene and quantify gene expression profile, and analyzed aflatoxin B1 levels, respectively. The results shown that all aflatoxigenic strains analyzed in this study are clustered within the same clade based on the aflD gene. The targeted aflD gene for expression was amplified in all aflatoxigenic strains with higher relative level in Aspergillus flavus but no relative expression shown in Aspergillus fumigatus. Enzyme-Linked Immunosorbent Assay showed that 43.1 % of the samples were found positive, with15.9 % being local rice and 27.2 % imported rice. Notably, 11.3 % of analyzed samples were above the recommended aflatoxin B1 in rice grains established by European Union with aflatoxin concentration ranging between 0 μg/kg to 3.2 μg/kg. These results are indicative of exposure of the population to aflatoxin B1 and possible health hazard. Our study reveals high levels of aflatoxin B1 contamination in both imported and local rice grains in Kenya. This study highlights the aflatoxigenic strains which are the potential aflatoxins contamination of different types of foodstuffs.

Occurrence of Alternaria Toxins, Ochratoxin A and Aflatoxins B1 in Green Arabic Coffee (Coffea arabica) in Saudi Arabia: Risk Assessment on HepG-2 Cell Line and Male Albino Rats

Arabica coffee (Coffee arabica L.) is among the most popular and widely consumed drinks globally today. The current study aimed to detect and quantify of potential mycotoxins in coffee beans and to investigate their potential risk on HepG-2 human liver cancer cells and male albino rats. Eighteen fully matured coffee beans samples were collected from different geographical locations in Saudi Arabia. Mycotoxins detected and quantified utilizing high-performance liquid chromatography (HPLC). Cytotoxicity of the detected toxins was investigated utilizing the MTT assay method toward HepG-2 cells and male albino rats. Cytotoxic properties of the detected mycotoxins were investigated at doses of 31.25, 62.50, 125, 250 and 500 µg/ml following exposure durations of 24 and 48 h in comparison with control. HPLC analysis revealed the detection of four Alternaria toxins (ALT-Ts) namely Altenuene (ALT), Tenuazonic acid (TA), Alternariol (AOH), and Altenuisol being AOH reported as the frequently detected mycotoxin in 18 coffee beans samples representing 75 % with quantity ranged 0.98 to 8.97 μg/g. OTA was frequently detected (10 samples, 55.5%) in concentrations varied from 0.75 to 9.87 μg/g. In addition, Aflatoxin B1 was also detected with low quantity ranged 0.89 to 3.22 μg/g. Treatment of HepG-2 cell line with combined Alternaria toxins (ALT, TeA, AOH, and Altenuisol), Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) as well as their combination triggered a decline in cell viability that was proportional to the dose administered. The elevated doses (62.50, 125, 250 and 500 µg/ml) notably reduced cell viability and induced cellular damages when contrasted with the lowest dose (31.25 µg/ml) and control. The IC50 for the tested mycotoxins registered for ALT-Ts, AFB1 and OTA as well as their combination were 115.95, 56.68, 59.86 and 31.05 µg/ml after 48 h of exposure. Secreting inflammatory markers (Interleukin-6 (IL-6), interleukin-1β (IL-1β) and COX-2), and oxidative markers (total antioxidant capacity (TAC), glutathione (GSH) and nitrite (NO) were downregulated by all mycotoxins with the highest decrease induced by their mixture. Malonaldehydes (MDA) and TAC were significantly increased by all toxins in treated rats. Hematological results revealed that AFB1, OTA and ALT-Ts-treated rats exhibited a notable decline in hemoglobin (Hb), red blood cells (RBCs), white blood cells (WBCs), and packed cell volume % (PCV). However, single and double dose-treated rats with OTA, Mix1 and Mix2 exhibited a marked increase in WBCs. The results demonstrated a notable decline in liver and kidney functions in all mycotoxins-treated rats. The studied organs (liver, kidney and spleen) exhibited significant damage, with the most severe alterations observed in the Mix1 and Mix2-treated group in comparison with the groups receiving individual treatment. This indicates that the mixture of AFB1, ALT-Ts and OTA had an enhanced toxic consequence against treated HepG-2 cell line and rats. Our results indicated the synergistic effect of the combined different mycotoxins against HepG-2 human liver cancer cells and male albino rats. This work could help fill in data gaps and enhance risk assessment for human health by addressing some of the possible risks associated with Alternaria toxins