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Aflatoxins, a group of toxic secondary metabolites produced by Aspergillus species, pose significant threats to human health due to their potent carcinogenic, mutagenic, and immunosuppressive properties. Chronic exposure to these contaminants, commonly found in staple foods such as maize and groundnuts, has been linked to an increased risk of liver cancer, growth impairment, and immune dysfunction. Several agents, such as calcium montmorillonite clay and Lactobacillus rhamnosus GG, have shown promise in reducing aflatoxin bioavailability and alleviating its toxic effects. Additionally, dietary supplements such as chlorophyllin, selenium, and N-acetylcysteine have demonstrated potential as adjuvants to counteract aflatoxin-induced oxidative stress and support liver function. In this treatise, some of the most discussed approaches to mitigating aflatoxin effects are explored in terms of their efficacy, safety, and potential mechanisms of action, which include direct aflatoxin binding, detoxification, cellular antioxidative, and hepatocellular protection properties. However, the effectiveness of these strategies can be influenced by various factors, such as dose, duration of exposure, and individual susceptibility. Therefore, further research is needed to optimize these interventions and develop new, targeted therapies for the prevention and treatment of aflatoxin-related diseases. This review aims to provide a comprehensive analysis of 18 pharmaceutical, nutraceutical, supplement, and probiotic strategies currently available for mitigating the deleterious effects of chronic aflatoxin exposure in humans and animal models. [ABSTRACT FROM AUTHOR]

Purpose of review: Aflatoxin B1 (AFB1) has been categorized as a Group I carcinogen by the International Agency for Research on Cancer (IARC). Considering its significant public health implications, it is crucial to have a comprehensive understanding of the different routes of exposure and biomarkers of aflatoxin. Recent findings: The main mechanisms by which aflatoxins cause harm to the body are oxidative stress, immune suppression, and DNA damage. More attention has recently been paid to investigating exposure through occupational and transplacental routes. Aflatoxin exposure is linked to unfavorable pregnancy outcomes, like low birth weight, according to the recent literature currently available on human studies. As aflatoxin can cause a variety of mutations and DNA damage, the AFB1-N7 guanine adduct has gained special interest in cancer research and toxicity. The most commonly used biomarker currently for determining chronic and consistent exposure to this carcinogen is aflatoxin B1-lysine serum albumin adduct in human blood samples (2–3 months). Recent research also investigates the potential for using hair analysis to study chronic exposure. Summary: The three main routes of aflatoxin exposure are dietary, occupational, and transplacental. Biomarkers can be found in a variety of biological samples, such as blood, hair, faeces, milk, urine, and bronchalveolar lavage. Surprisingly, there is an absence of cutoff limits for aflatoxin levels in biological fluids concerning adverse health effects in humans. Given the potential carcinogenic effects and the significant public health implications of aflatoxin, prioritizing the identification of new biomarkers for evaluating chronic exposure is crucial. [ABSTRACT FROM AUTHOR]

This study aimed to diagnose the ability of two probiotic strains isolated from Siahmazgi cheese to reduce the concentration of aflatoxin M1 (AFM1), which was able to reduce it up to 50%. Two strains of Lactobacillus isolated from local Siahmazgi cheese, were inoculated into raw milk containing 10 ng/mL AFM1. AFM1 levels were assessed by HPLC at 0, 24, 48 and 72 h. Also, for the assessment of the possible reduction of AFM1 by non-viable bacterial cells, heat treatment was applied in milk. In general, significant differences were observed between the control group and AFM1 contaminated milk samples containing probiotics during 72 h of refrigeration (p < 0.05). In the presence of Lactobacillus brevis, Lactobacillus plantarum and mixture probiotics, the concentrations of AFM1 ranged from 0.102 ± 0.004 ng/mL to 0.052 ± 0.002 ng/mL, 0.114 ± 0.002 ng/mL to 0.074 ± 0.003 ng/mL and 0.108 ± 0.003 ng/mL to 0.065 ± 0.002 ng/mL, respectively. Also, in the heated group, just a decrease was observed in the first 24 h. The results showed that native Iranian strains of Lactobacillus have good potential in reducing AFM1 in milk. The time factor is effective in reducing AFM1 by living bacterial cells. [ABSTRACT FROM AUTHOR]

Oral adsorbents have attracted increasing attention as a potential therapeutic strategy for the removal of toxic substances from the gastrointestinal tract. This review aims to provide a comprehensive overview of the structural and mechanic properties of oral adsorbents such as activated charcoal, medical clays, zeolites, silica-based adsorbents, dietary fibers (carbohydrates alginate, chitosan, pectin, carrageenan etc.), and their physicochemical properties. The review also explores the mechanisms of action of oral adsorbents, including physical adsorption, ion exchange, and specific binding interactions. Furthermore, we provide an overview of the factors that influence the adsorption capacity and selectivity of oral adsorbents. The summary of recent advances in the usage of thiolated adsorbents is discussed, which provides valuable insights into the structural and mechanistic properties of oral adsorbents and their potential as a therapeutic option for the removal of toxic substances from the gastrointestinal tract. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the effect of bay leaf (Syzygium polyanthum) infusion on preserving the quality of corn during storage. The research employed an experimental approach using a Completely Randomized Design (CRD) with five treatments and four replications. The treatments included A (control corn), B (25% (v/v) bay leaf infusion), C (50% (v/v) bay leaf infusion), D (75% (v/v) bay leaf infusion), and E (100% (v/v) bay leaf infusion). The observed variables included damaged kernels, moldy kernels, moisture content, and aflatoxin content. Corn with high aflatoxin content was immersed in bay leaf infusion according to the respective treatments and then stored for four weeks. The research results showed that soaking corn with high aflatoxin content in bay leaf infusion (had a significant effect (P<0.01) on moisture and aflatoxin content but did not have a considerable impact (P>0.05) on damaged kernels and moldy kernels. Based on the research findings, it was concluded that the application of 100% (v/v) bay leaf infusion improved the quality of corn during a 4 -week storage period, with the percentage of damaged kernels at 4.75%, moldy grains at 0.75%, moisture content at 14.60%, and a decrease in aflatoxin content by 41.11% (150 -88.33 ppb). [ABSTRACT FROM AUTHOR]

Aspergillus flavus is a cosmopolitan saprophytic fungus that infests several foodstuffs and is associated with adverse effects in humans. In Senegal, significant losses of groundnut production are mainly due to contamination caused by this species. This study evaluated in vitro antifungal activities of Cyperus articulatus, Cyperus rotundus and Lippia alba essential oils against A. flavus isolated from peanut seeds. Essential oils obtained by hydrodistillation of rhizomes of the two Cyperus species and leaves of L. alba were analyzed with GC-DIF and GC-MS. The essential oil yields from C. articulatus, C. rotundus and L. alba were 1.1%, 1.3% and 1.7%, respectively. These three samples had the following chemotypes: (i) mustakone (21.4%)/eudesma-4(15)-7-dien-1β-ol (8.8%)/caryophyllene oxide (5.9%), (ii) caryophyllene oxide (25.2%)/humulene epoxyde 2 (35.0%) and (iii) geranial (46.6%)/neral (34.6%). The three oils tested inhibited the growth of A. flavus at concentrations between 100 and 1000 ppm. The L. alba oil was the most effective with total clearance of A. flavus on PDA. For the essential oils of C. rotundus (93.65%) and C. articulatus (78.11%), the highest inhibition rates were obtained with a 1000 ppm dose. Thus, L. alba oil could be used safely as an effective protector of groundnuts against A. flavus. [ABSTRACT FROM AUTHOR]

Approximately 25% of cereal grains present with contamination caused by fungi and the presence of mycotoxins that may cause severe adverse effects when consumed. Maize has been genetically engineered to present different traits, such as fungal or insect resistance and herbicide tolerance. This systematic review compared the observable quantities, via meta-analysis, of four mycotoxins (aflatoxins—AFL, fumonisins—FUM, deoxynivalenol—DON, zearalenone—ZEA) between genetically modified (GM) and conventional maize kernels. This study was conducted following the PRISMA guidelines, with searches performed using PubMed, Web of Science, Scopus, Google Scholar, and CAPES journals databases. Analyses were conducted using RevMan v.5.4 software. Transgenic maize showed a 58% reduction in total mycotoxins (p < 0.001) compared to conventional maize. FUM were the most impacted, with a 59% reduction (p < 0.001) in GM maize. AFL and ZEA levels were also lower in GM maize by 49% (p = 0.02) and 51% (p < 0.001), respectively. On the other hand, DON levels increased by 6% (p < 0.001) in GM maize compared to conventional maize. However, results for ZEA and DON were inconclusive due to the limited research and sample sizes. We conclude that transgenic maize reduces total mycotoxins by over 50%, primarily fumonisin and aflatoxin. Most studies presented maize varieties that were resistant to insects or herbicides, not fungal pathogens, showing a positive collateral effect of these genetic alterations. Therefore, transgenic maize appears to be a safer product for animal and human consumption from a toxicological point of view. Further studies with larger sample sizes are needed to confirm our findings for ZEA and DON in transgenic maize. [ABSTRACT FROM AUTHOR]

The climate-change-coupled fungal burden in crop management and the need to reduce chemical pesticide usage highlight the importance of finding sustainable ways to control Aspergillus flavus. This study examines the effectiveness of 50 Pseudomonas isolates obtained from corn rhizospheres against A. flavus in both solid and liquid co-cultures. The presence and quantity of aflatoxin B1 (AFB1) and AFB1-related compounds were determined using high-performance liquid chromatography–high resolution mass spectrometry analysis. Various enzymatic- or non-enzymatic mechanisms are proposed to interpret the decrease in AFB1 production, accompanied by the accumulation of biosynthetic intermediates (11-hydroxy-O-methylsterigmatocystin, aspertoxin, 11-hydroxyaspertoxin) or degradation products (the compounds C16H10O6, C16H14O5, C18H16O7, and C19H16O8). Our finding implies the upregulation or enhanced activity of fungal oxidoreductases and laccases in response to bacterial bioactive compound(s). Furthermore, non-enzymatic reactions resulted in the formation of additional degradation products due to acid accumulation in the fermented broth. Three isolates completely inhibited AFB1 or any AFB1-related compounds without significantly affecting fungal growth. These bacterial isolates supposedly block the entire pathway for AFB1 production in the fungus during interaction. Apart from identifying effective Pseudomonas isolates as potential biocontrol agents, this work lays the foundation for exploring new bacterial bioactive compounds. [ABSTRACT FROM AUTHOR]

Food crops around the world are commonly contaminated with Aspergillus flavus, which can produce the carcinogenic mycotoxin aflatoxin B1 (AFB1). The objective of this study is to test an X-ray irradiation sterilization method for studying AFB1 in contaminated maize samples in the laboratory. Maize that had been naturally contaminated with 300 ppb AFB1 by the growth of aflatoxigenic A. flavus was ground and then irradiated at 0.0, 1.0, 1.5, 2.0, 2.5, and 3.0 kGy. A. flavus was quantified by dilution plating on potato dextrose agar (PDA) and modified Rose Bengal media (MDRB) for viability and qPCR for gene presence. AFB1 was quantified by HPLC and ELISA. A. flavus viability, but not gene copies, significantly decreased with increasing doses of radiation (PDA: p < 0.001; MDRB: p < 0.001; qPCR: p = 0.026). AFB1 concentration did not significantly change with increasing doses of radiation (HPLC: p = 0.153; ELISA: p = 0.567). Our results imply that X-ray irradiation is an effective means of reducing viable A. flavus without affecting AFB1 concentrations. Reducing the hazard of fungal spores and halting AFB1 production at the targeted dose are important steps to safely and reproducibly move forward research on the global mycotoxin challenge. [ABSTRACT FROM AUTHOR]

AbstractAflatoxins are carcinogens that can contaminate food and affect various body organs especially liver and kidney. When consumed, aflatoxin B1 (AFB1) is partially metabolised into aflatoxin M1 (AFM1), which is excreted in the urine.Breast milk may also contain AFM1 due to maternal dietary intake from contaminated food. This cross-sectional study aimed to determine the levels of AFM1 in both urine and breast milk among breastfeeding mothers (n = 256). The mother’s demographic information was collected during recruitment. Mothers were then scheduled for an appointment to provide a morning urine sample along with five to ten mL samples of breast milk. AFM1 levels in both samples were analysed using an enzyme-linked immunosorbent assay (ELISA). Spearman’s rho and Chi-square were used to determine the associations between mean levels of AFM1 in urine and breast milk. Findings show 68.0% of urine samples were contaminated with AFM1 (mean levels = 0.08 ± 0.04 ng/mL), while 14.8% of breast milk samples had AFM1 (mean levels = 5.94 ± 1.81 ng/kg). Urine AFM1 levels were not significantly associated with AFM1 levels in breast milk (p > 0.05). This study can act as a baseline for future research examining long-term aflatoxin exposure among both mothers and infants. [ABSTRACT FROM AUTHOR]

Aspergillus section Flavi (Flavi) is a diverse group of fungal species whose common members include A. flavus and A. parasiticus. These are well-known for the production of aflatoxin (AF) B and G and other toxic metabolites, like cyclopiazonic acid (CPA). They are saprophytic soil dwellers and also become crop opportunistic epiphytes. The consequence is contamination of the crop with mycotoxins, such as carcinogenic AF. We investigated the Flavi community structure of maize and that of their surrounding soil, including their mycotoxigenicity. Furthermore, we investigated the link of the maize Flavi diversity with preharvest maize AF levels. The study was carried out in four selected districts of Zambia, in a low rainfall zone. The Flavi characterisation was triphasic, involving morphological (colony colour and sclerotia formation), metabolic (AF and CPA production) and genetic (calmodulin gene polymorphism) analyses. Flavi abundance was determined by dilution plate technique on modified rose Bengal agar. Results showed that Flavi communities on maize and in soil differed. Maize had a higher Flavi species diversity than soil. A. parasiticus dominated the soil community by frequency of field appearance (85%), while maize was dominated by A. minisclerotigenes (45%). CPA-producers with or without AF production dominated the maize (65%) while producers of only AF (B/G) dominated the soil (88%). The ratio between maize A. parasiticus and A. minisclerotigenes abundance seemed to have had a bearing on the levels of AF in maize, with a ratio close to 1:1 having higher levels than a pure community of either A. parasiticus or A. minisclerotigenes. [ABSTRACT FROM AUTHOR]

Aspergillus flavus produces aflatoxin, a carcinogenic fungal toxin that poses a threat to the agricultural and food industries. There is a concern that the distribution of aflatoxin-producing A. flavus is expanding in Japan due to climate change, and it is necessary to understand what types of strains inhabit. In this study, we sequenced the genomes of four Aspergillus strains isolated from agricultural fields in the Ibaraki prefecture of Japan and identified their genetic variants. Phylogenetic analysis based on single-nucleotide variants revealed that the two aflatoxin-producing strains were closely related to A. flavus NRRL3357, whereas the two non-producing strains were closely related to the RIB40 strain of Aspergillus oryzae, a fungus widely used in the Japanese fermentation industry. A detailed analysis of the variants in the aflatoxin biosynthetic gene cluster showed that the two aflatoxin-producing strains belonged to different morphotype lineages. RT-qPCR results indicated that the expression of aflatoxin biosynthetic genes was consistent with aflatoxin production in the two aflatoxin-producing strains, whereas the two non-producing strains expressed most of the aflatoxin biosynthetic genes, unlike common knowledge in A. oryzae, suggesting that the lack of aflatoxin production was attributed to genes outside of the aflatoxin biosynthetic gene cluster in these strains. [ABSTRACT FROM AUTHOR]

Non-genetic variation limits the identification of novel maize germplasm with genetic markers for reduced Aspergillus flavus infection and aflatoxin contamination. Aflatoxin measurements can vary substantially within fields containing the same germplasm following inoculation with A. flavus. While some variation is expected due to microenvironmental differences, components of field screening methodologies may also contribute to variability in collected data. Therefore, the objective of this study is to test the effects of three different shelling methods (whole ear (WE), ear end removal (EER), and inoculation site-surrounding (ISS)) to obtain bulk samples from maize on aflatoxin measurements. Five ears per row of three inbred lines and two hybrids were inoculated with A. flavus, then shelled using the three different methods, and aflatoxin was quantified. Overall, EER and ISS resulted in reduced coefficients of variance (CVs) in comparison to WE for both inbred and hybrid maize lines, with two exceptions. Susceptible B73 showed increased CVs with both EER and ISS compared to WE, and resistant Mp719's EER CVs marginally increased compared to WE. While WE is the standard practice for most breeding programs due to its technical simplicity, EER and ISS may allow for finely phenotyping parental lines for further breeding applications. [ABSTRACT FROM AUTHOR]

Chemical pesticides help reduce crop loss during production and storage. However, the carbon footprints and ecological costs associated with this strategy are unsustainable. Here, we used three in vitro models to characterize how different Trichoderma species interact with two aflatoxin producers, Aspergillus flavus and Aspergillus parasiticus, to help develop a climate-resilient biological control strategy against aflatoxigenic Aspergillus species. The growth rate of Trichoderma species is a critical factor in suppressing aflatoxigenic strains via physical interactions. The dual plate assay suggests that Trichoderma mainly suppresses A. flavus via antibiosis, whereas the suppression of A. parasiticus occurs through mycoparasitism. Volatile organic compounds (VOCs) produced by Trichoderma inhibited the growth of A. parasiticus (34.6 ± 3.3%) and A. flavus (20.9 ± 1.6%). The VOCs released by T. asperellum BTU and T. harzianum OSK-34 were most effective in suppressing A. flavus growth. Metabolites secreted by T. asperellum OSK-38, T. asperellum BTU, T. virens OSK-13, and T. virens OSK-36 reduced the growth of both aflatoxigenic species. Overall, T. asperellum BTU was the most effective at suppressing the growth and aflatoxin B1 production of both species across all models. This work will guide efforts to screen for effective biological control agents to mitigate aflatoxin accumulation. [ABSTRACT FROM AUTHOR]

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Aflatoxin intoxication has been reported in most species; however, it is infrequently documented in sheep. In this report, three 7-month-old Romane ewe lambs were found with severe icterus, tachypnea, slight fever, diarrhea, anorexia, and lethargy. Two of them were culled, and one recovered after treatment. Necropsy findings showed jaundice, ascites, diffuse hemorrhage on the peritoneum, gall bladder enlargement, and alteration in the color and texture of the liver and kidney. Histopathological evaluation revealed hepatocellular necrosis, megalocytosis, centrilobular to bridging fibrosis in the liver, and hemoglobinuria without nephrotic degeneration in the kidney. The polymerase chain reaction test for leptospirosis as the main differential diagnosis was conducted, and the result was negative. Similarly, the Warthin Starry special staining for spirochetes was negative. Clinical signs and characteristics of microscopic lesions were suggestive of aflatoxicosis in the lambs described in this investigation. As sheep flocks with intensive rearing systems are becoming more common in Iran, the emergence of aflatoxicosis due to fungi-contaminated feedstuff can be a serious health concern in such farms. [ABSTRACT FROM AUTHOR]

BackgroundAflatoxin (AF) exposure is associated with child growth faltering in cross-sectional studies, with limited findings from longitudinal studies.ObjectivesTo evaluate the relationship between maternal AF B1-lysine adduct concentration, child AF B1-lysine adduct concentration, and child growth in the first 30 mo of life.MethodsAF B1-lysine adduct was measured in mother-child dyad plasma samples using isotope dilution mass spectrometry. Using linear regression, we assessed the relationship between AF B1-lysine adduct concentration and child weight, height, and head and mid-upper arm circumferences at 1 wk, 6, 12, 18, 24, and 30 mo of age.ResultsIn adjusted models, maternal prenatal AF B1-lysine adduct (pg/μL) was positively associated with newborn anthropometric outcomes; largest beta coefficients for associations between standardized values were for newborn weight-for-age z-score [β = 0.13; 95% confidence interval (CI): 0.02, 0.24; P < 0.05 and β = 0.11; 95% CI: 0.00, 0.22; P < 0.05 for second and third trimester AF, respectively]. Child AF B1-lysine adduct (pg/μL) at 6 mo was negatively associated with head circumference-for-age z-score at 6, 18, 24, and 30 mo, with beta coefficients ranging from β = -0.15; 95% CI: -0.28, -0.02 to β = -0.17; 95% CI: -0.31, -0.03; P < 0.05); 18-mo AF was negatively associated with anthropometric outcomes at 18, 24, and 30 mo, most consistently with length-for-age z-score (β = -0.18; 95% CI: -0.32, -0.04, β = -0.21; 95% CI: -0.35, -0.07, β = -0.18; 95% CI: -0.32, -0.03 at 18, 24 and 30 mo, respectively).ConclusionsChild AF exposure was associated with impaired child growth, but maternal AF exposure was not. Exposure during infancy was linked to persistent deficit in head circumference, implying reduced brain size lasting beyond the age of 2 years. Exposure at 18 mo was linked to persistent linear growth deficit. Further research should elucidate mechanisms through which AF affects child growth.

Aflatoxins (AFs) are highly toxic mycotoxins produced by the fungi, Aspergillus flavus and Aspergillus parasiticus. AFs pose severe health risks owing to their acute toxicity and carcinogenic properties. The control of AF contamination remains significantly challenging despite the extensive efforts toward controlling it. Here, we investigated the potential of mushroom extracts as a source of AF biosynthetic inhibitors. The A. parasiticus mutant strain, NFRI-95, that accumulates an AF biosynthesis intermediate, norsolorinic acid, was used in the bioassay to detect the inhibitory activity against AF biosynthesis. The screening of 195 mushroom extracts revealed that the culture filtrate extract of Chondrostereum purpureum exhibited strong inhibitory activity against AF biosynthesis. Next, large-scale culturing of C. purpureum was performed to isolate the compounds accounting for the inhibitory activity. The culture filtrate was extracted with ethyl acetate, after which the active compound was isolated by silica gel column chromatography and preparative high performance liquid chromatography (HPLC). The active compound was identified as cyclo(Val–Pro) by spectroscopic analyses. Further, four stereoisomers of cyclo(Val–Pro) were synthesized by the condensation of the N -Boc derivatives of d - and l -valine with the methyl esters of d - and l -proline. The naturally isolated compound was identified as cyclo(l -Val- l -Pro) by comparing its retention time with those of synthetic compounds by chiral HPLC analysis and CD spectra. The IC 50 value of cyclo(L-Val-L-Pro) was 2.4 mM, whereas the LD, DL, and DD isomers exhibited weaker activities, with IC 50 values of >5 mM. • Aflatoxin (AF) biosynthesis inhibitors were explored using mushroom extracts. • An extract of Chondrostereum purpureum effectively inhibited AF biosynthesis. • Cyclo(l -valyl- l -proline) was isolated from the C. purpureum culture filtrate. • Four cyclo(Val–Pro) isomers were synthesized. • Cyclo(l -valyl- l -proline) showed the strongest inhibitory activity among the isomers. [ABSTRACT FROM AUTHOR]

Aflatoxin B1 (AFB1) is a plant-origin toxin that could induce oxidative stress in fish. The micromineral selenium (Se) possesses well-documented antioxidant properties. To assess the ameliorative effects of SeNPs (1 mg/kg fish feed) on oxidative stress induced by AFB1 (500 μg/kg fish feed), Nile tilapia (32.2±1.7 g body weight) were distributed randomly and even in six groups for 8-week feeding trial. Live enzymes, AST, ALT, and ALP levels were increased in the serum of fish fed AFB1-contaminated diet, and the addition of SeNPs could restore normal values compared to the control. The gene expression of antioxidant enzymes, superoxide dismutase (SOD) enzyme and catalase (CAT) enzyme, and DNA fragmentation were significantly increased in response to aflatoxin exposure, while dietary SeNPs could mitigate the generated oxidative stress. The innate immunity, serum antibacterial activity (SAA), oxidative burst activity (OBA), phagocytic activities (PA and PI), and gene expression of cytokines (interleukin (IL)-1β, heat shock protein70 (Hsp), and tumor necrosis factor (TNF)-α) revealed a status of immunosuppression in Nile tilapia fed on AFB1-contaminated diet. These findings showed that fish became more vulnerable to Streptococcus agalactiae infection with a high mortality rate while dietary SeNPs provided a high relative protection level (RPL). From the obtained findings, SeNPs could mitigate the oxidative stress induced by feeding the AFB1 diet and could boost the immunity of stressed Nile tilapia. [ABSTRACT FROM AUTHOR]

This study was conducted in the Plant Pathology Laboratory/ College of Agriculture/ University of Anbar to study and test the effectiveness of plant extracts (turmeric, pomegranate peel, banana peel) in inhibiting the growth of the fungus Aspergillus flavus and test its ability to reduce aflatoxin B1 from liquid media. The plant materials (powders and extracts) were used in three concentrations (10, 15, 20) mg ml-1. The results showed that all plant materials were significantly effective in inhibiting the growth of A. flavus and their ability to reduce aflatoxin B1 from liquid media. Turmeric powder showed its superiority in its inhibitory effect on the growth of fungi on PDA culture medium, as it achieved the highest percentage of inhibition at a concentration of 20 mg ml-1, which amounted to 87.41%. Whereas, banana peel powder gave the lowest inhibition rate at 10mg ml-1 concentration) 64.07% (. While the aqueous extract of turmeric achieved inhibition in the growth of the fungus at a concentration of 20 mg ml-1 by 87.41%, outperformed the aqueous extract of pomegranate and banana, where the rates of inhibition reached 86.66% and 82.96%, respectively at the same concentration. In addition, the alcoholic extract of turmeric showed the highest rate of inhibition of fungi growth at a concentration of 20 mg ml-1, which amounted to 94.07%, followed by the alcoholic extract of banana and pomegranate with a percentage of 93.3% and 88.14%, respectively, at the same concentration.

Abstract Aflatoxin, mainly produced by Aspergillus flavus, is one of the most notorious mycotoxin for its toxicity and carcinogenicity. Despite extensive efforts, effective strategies to control A. flavus and AFB1 contamination remain elusive. Here, we investigate the potential of p‐anisaldehyde (AS), an aldehyde derived from plant essential oils, as a natural antifungal agent against A. flavus and its ability to modulate AFB1 biosynthesis. We found that AS exhibited broad‐spectrum antifungal activities against Aspergillus spp. and effectively inhibited A. flavus asexual development, AFB1 production, and pathogenicity. AS treatment disrupted the cell surface structure and membrane integrity, as observed by scanning electron microscopy and PI staining. RNA‐sequencing analysis revealed that AS significantly altered the expression of genes involved in redox homeostasis, plasma membrane function, and cell cycle progression. Further investigation demonstrated that AS induced a reduction in mitochondrial membrane potential (Δψm) and accumulation of reactive oxygen species (ROS), leading to cell cycle arrest at the G2/M phase. The AS‐induced ROS accumulation was found to be mitigated by the superoxide dismutase‐mediated antioxidant system, which is regulated by transcriptional factor Ap1. Notably, the Ap1‐regulatory ROS detoxification system was also found to be involved in A. flavus pathogenicity and AFB1 production. Overall, these findings provide valuable insights into the inhibitory mechanism of AS against A. flavus, paving the way for its potential application as a natural strategy to mitigate AFB1 contamination in both food and agriculture crops.

Abstract Maize (Zea mays), a major food crop worldwide, is susceptible to infection by the saprophytic fungus Aspergillus flavus that can produce the carcinogenic metabolite aflatoxin (AF) especially under climate change induced abiotic stressors that favor mold growth. Several studies have used “-omics” approaches to identify genetic elements with potential roles in AF resistance, but there is a lack of research identifying the involvement of small RNAs such as microRNAs (miRNAs) in maize-A. flavus interaction. In this study, we compared the miRNA profiles of three maize lines (resistant TZAR102, moderately resistant MI82, and susceptible Va35) at 8 h, 3 d, and 7 d after A. flavus infection to investigate possible regulatory antifungal role of miRNAs. A total of 316 miRNAs (275 known and 41 putative novel) belonging to 115 miRNA families were identified in response to the fungal infection across all three maize lines. Eighty-two unique miRNAs were significantly differentially expressed with 39 miRNAs exhibiting temporal differential regulation irrespective of the maize genotype, which targeted 544 genes (mRNAs) involved in diverse molecular functions. The two most notable biological processes involved in plant immunity, namely cellular responses to oxidative stress (GO:00345990) and reactive oxygen species (GO:0034614) were significantly enriched in the resistant line TZAR102. Coexpression network analysis identified 34 hubs of miRNA-mRNA pairs where nine hubs had a node in the module connected to their target gene with potentially important roles in resistance/susceptible response of maize to A. flavus. The miRNA hubs in resistance modules (TZAR102 and MI82) were mostly connected to transcription factors and protein kinases. Specifically, the module of miRNA zma-miR156b-nb – squamosa promoter binding protein (SBP), zma-miR398a-3p – SKIP5, and zma-miR394a-5p – F-box protein 6 combinations in the resistance-associated modules were considered important candidates for future functional studies.

Abstract Background Aspergillus flavus is an important agricultural and food safety threat due to its production of carcinogenic aflatoxins. It has high level of genetic diversity that is adapted to various environments. Recently, we reported two reference genomes of A. flavus isolates, AF13 (MAT1-2 and highly aflatoxigenic isolate) and NRRL3357 (MAT1-1 and moderate aflatoxin producer). Where, an insertion of 310 kb in AF13 included an aflatoxin producing gene bZIP transcription factor, named atfC. Observations of significant genomic variants between these isolates of contrasting phenotypes prompted an investigation into variation among other agricultural isolates of A. flavus with the goal of discovering novel genes potentially associated with aflatoxin production regulation. Present study was designed with three main objectives: (1) collection of large number of A. flavus isolates from diverse sources including maize plants and field soils; (2) whole genome sequencing of collected isolates and development of a pangenome; and (3) pangenome-wide association study (Pan-GWAS) to identify novel secondary metabolite cluster genes. Results Pangenome analysis of 346 A. flavus isolates identified a total of 17,855 unique orthologous gene clusters, with mere 41% (7,315) core genes and 59% (10,540) accessory genes indicating accumulation of high genomic diversity during domestication. 5,994 orthologous gene clusters in accessory genome not annotated in either the A. flavus AF13 or NRRL3357 reference genomes. Pan-genome wide association analysis of the genomic variations identified 391 significant associated pan-genes associated with aflatoxin production. Interestingly, most of the significantly associated pan-genes (94%; 369 associations) belonged to accessory genome indicating that genome expansion has resulted in the incorporation of new genes associated with aflatoxin and other secondary metabolites. Conclusion In summary, this study provides complete pangenome framework for the species of Aspergillus flavus along with associated genes for pathogen survival and aflatoxin production. The large accessory genome indicated large genome diversity in the species A. flavus, however AflaPan is a closed pangenome represents optimum diversity of species A. flavus. Most importantly, the newly identified aflatoxin producing gene clusters will be a new source for seeking aflatoxin mitigation strategies and needs new attention in research.

ABSTRACT Aflatoxin, produced by Aspergillus flavus and Aspergillus paraciticus, commonly contaminates agricultural and processed products. This study aims to analyze aflatoxin levels in fried onions produced in Palu through high-performance liquid chromatography (HPLC) using an immunoaffinity column an a fluorescent detector at an excitation wavelength of 360 nm and an emission wavelength of 440 nm. The standard working chromatogram of aflatoxin gave a peak response at retention times of 28.618 minutes for aflatoxin B1, 21.985 minutes for aflatoxin B2, 17.625 minutes for aflatoxinn G1, and 13.719 minutes for aflatoxin G2. The limit of detection (LOD) values for each standard were : aflatoxin B1 : 0.2607 ng/ml; aflatoxin B2 : 0.0600 ng/ml; aflatoxin G1 : 0.1986 ng/ml; and aflatoxin G2 : 0.0310 ng/ml. The chromatogram results of the 10 fried onion samples analyzed showed no peak response with the same retention time as the standard aflatoxin retention time, so that the 10 samples can be concluded as meet requirements. Keywords: Aflatoxin, HPLC, Immunoaffinity colum, P Palu fried shallot

Based on simulating the preparation of aged diet with excessive aflatoxin, the effects of aged diet on the feed-meat ratio, immune organ indexes and blood cell count of chicken, and the recovery effect of Lactobacillus sp. were studied. Thirty-six white Laihang Chicken were randomly divided into 4 groups: control group, high-dose aflatoxin group, low-dose aflatoxin group, high-dose aflatoxin+Lactobacillus sp. group, the test period was 28 days. The results showed that compared with the control group, aflatoxin had a significant retardation effect on the daily weight gain of chicken. Compared with the control group, the feed-meat ratio in the low-dose aflatoxin group and high-dose aflatoxin group significantly increased. Adding Lactobacillus sp. had a significant improvement effect on the feed-meat ratio (P<0.05). Compared with the control group, aflatoxin had a significant retardation effect on the formation of chicken blood cells and the development of immune organs (P<0.05), resulting in the blocked synthesis of blood cells and delayed development of immune organs, while Lactobacillus sp. had some recovery effects on the formation of chicken blood cells and the development of immune organs. [ABSTRACT FROM AUTHOR]

Aims The present work aimed to distinguish the indigenous Aspergillus flavus isolates obtained from the first (pioneer) grain corn farms in Terengganu, Malaysia, into aflatoxigenic and non-aflatoxigenic by molecular and aflatoxigenicity analyses, and determine the antagonistic capability of the non-aflatoxigenic isolates against aflatoxigenic counterparts and their aflatoxin production in vitro. Methods and results Seven A. flavus isolates previously obtained from the farms were characterized molecularly and chemically. All isolates were examined for the presence of seven aflatoxin biosynthesis genes, and their aflatoxigenicity was confirmed using high performance liquid chromatography with fluorescence detector. Phylogenetic relationships of all isolates were tested using ITS and β-tubulin genes. Of the seven isolates, two were non-aflatoxigenic, while the remaining were aflatoxigenic based on the presence of all aflatoxin biosynthesis genes tested and the productions of aflatoxins B1 and B2. All isolates were also confirmed as A. flavus following phylogenetic analysis. The indigenous non-aflatoxigenic isolates were further examined for their antagonistic potential against aflatoxigenic isolates on 3% grain corn agar. Both non-aflatoxigenic isolates significantly reduced AFB1 production of the aflatoxigenic isolates. Conclusion The indigenous non-aflatoxigenic A. flavus strains identified in the present work were effective in controlling the aflatoxin production by the aflatoxigenic A. flavus isolates in vitro and can be utilized for in situ testing. [ABSTRACT FROM AUTHOR]

Aspergillus flavus is notorious for contaminating food with its secondary metabolite—highly carcinogenic aflatoxins. In this study, we found that exogenous nitric oxide (NO) donor could influence aflatoxin production in A. flavus. Flavohemoglobins (FHbs) are vital functional units in maintaining nitric oxide (NO) homeostasis and are crucial for normal cell function. To investigate whether endogenous NO changes affect aflatoxin biosynthesis, two FHbs, FHbA and FHbB, were identified in this study. FHbA was confirmed as the main protein to maintain NO homeostasis, as its absence led to a significant increase in intracellular NO levels and heightened sensitivity to SNP stress. Dramatically, FHbA deletion retarded aflatoxin production. In addition, FHbA played important roles in mycelial growth, conidial germination, and sclerotial development, and response to oxidative stress and high-temperature stress. Although FHbB did not significantly impact the cellular NO level, it was also involved in sclerotial development, aflatoxin synthesis, and stress response. Our findings provide a new perspective for studying the regulatory mechanism of the development and secondary mechanism in A. flavus. [ABSTRACT FROM AUTHOR]

Fungal infections were observed on the coffee plant, this study aims to determine the types of fungi causing this pollution and the possibility of producing mycotoxins or not. so we make this study to isolate and identify the contaminating fungi that find in three types of Arabica coffee plants, which are common in the city of Jazan in the Kingdom of Saudi Arabia, namely (Khawlani, Luqmati and Harari) by using the dilution method on Potato Dextrose Agar media (PDA). The results showed that some coffee samples were contaminated with fungi, which belong to 5 fungal genera, most of which belong to the Ascomycetes fungi. The species Aspergillus niger had the highest percentage of appearance in the coffee samples, followed by Aspergillus flavus and then Penicillium sp. The ammonia detection results showed that most Aspergillus flavus and A. ochrous isolates produced mycotoxins, while all isolates of Aspergillus niger and Penicillium sp. do not produce aflatoxins. [ABSTRACT FROM AUTHOR]

Foodborne diseases can be attributed not only to contamination with bacterial or fungal pathogens but also their associated toxins. Thus, to maintain food safety, innovative decontamination techniques for toxins are required. We previously demonstrated that an atmospheric-pressure dielectric-barrier discharge (APDBD) plasma generated by a roller conveyer plasma device is effective at inactivating bacteria and fungi in foods. Here, we have further examined whether the roller conveyer plasma device can be used to degrade toxins produced by foodborne bacterial pathogens, including aflatoxin, Shiga toxins (Stx1 and Stx2), enterotoxin B and cereulide. Each toxin was spotted onto an aluminum plate, allowed to dry, and then treated with APDBD plasma applied by the roller conveyer plasma device for different time periods. Assessments were conducted using a competitive enzyme-linked immunosorbent assay (ELISA) and liquid chromatography–tandem mass spectrometry (LC-MS/MS). The results demonstrate a significant time-dependent decrease in the levels of these toxins. ELISA showed that aflatoxin B1 concentrations were reduced from 308.6 µg/mL to 74.4 µg/mL within 1 min. For Shiga toxins, Stx1 decreased from 913.8 µg/mL to 65.1 µg/mL, and Stx2 from 2309.0 µg/mL to 187.6 µg/mL within the same time frame (1 min). Enterotoxin B levels dropped from 62.67 µg/mL to 1.74 µg/mL at 15 min, and 1.43 µg/mL at 30 min, but did not display a significant decrease within 5 min. LC-MS/MS analysis verified that cereulide was reduced to below the detection limit following 30 min of APDBD plasma treatment. Taken together, these findings highlight that a range of foodborne toxins can be degraded by a relatively short exposure to plasma generated by an APDBD using a roller conveyer device. This technology offers promising advancements in food safety, providing a novel method to alleviate toxin contamination in the food processing industry. [ABSTRACT FROM AUTHOR]

Copyright of Anbar Journal of Agricultural Sciences is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Aims: The current review aims to outline and summarize the latest research on aflatoxin, with research studies describing natural, herbal and chemical compound applications in animal (pig) models and in vitro cellular studies. Aflatoxin, a carcinogenic toxin metabolite, is produced by Aspergillus flavus in humid environments, posing a threat to human health and crop production. The current treatment involves the prevention of exposure to aflatoxin and counteracting its harmful toxic effects, enabling survival and research studies on an antidote for aflatoxin. Objectives: To summarize current research prospects and to outline the influence of aflatoxin on animal forage in farm production, food and crop processing. The research application of remedies to treat aflatoxin is undergoing development to pinpoint biochemical pathways responsible for aflatoxin effects transmission and actions of treatment. Significance: To underline the environmental stress of aflatoxin on meat and dairy products; to describe clinical syndromes associated with aflatoxicosis on human health that are counteracted with proposed treatment and preventive interventions. To understand how to improve the health of farm animals with feed conditions. [ABSTRACT FROM AUTHOR]

Maize (Zea mays), a major food crop worldwide, is susceptible to infection by the saprophytic fungus Aspergillus flavus that can produce the carcinogenic metabolite aflatoxin (AF) especially under climate change induced abiotic stressors that favor mold growth. Several studies have used "-omics" approaches to identify genetic elements with potential roles in AF resistance, but there is a lack of research identifying the involvement of small RNAs such as microRNAs (miRNAs) in maize-A. flavus interaction. In this study, we compared the miRNA profiles of three maize lines (resistant TZAR102, moderately resistant MI82, and susceptible Va35) at 8 h, 3 d, and 7 d after A. flavus infection to investigate possible regulatory antifungal role of miRNAs. A total of 316 miRNAs (275 known and 41 putative novel) belonging to 115 miRNA families were identified in response to the fungal infection across all three maize lines. Eighty-two unique miRNAs were significantly differentially expressed with 39 miRNAs exhibiting temporal differential regulation irrespective of the maize genotype, which targeted 544 genes (mRNAs) involved in diverse molecular functions. The two most notable biological processes involved in plant immunity, namely cellular responses to oxidative stress (GO:00345990) and reactive oxygen species (GO:0034614) were significantly enriched in the resistant line TZAR102. Coexpression network analysis identified 34 hubs of miRNA-mRNA pairs where nine hubs had a node in the module connected to their target gene with potentially important roles in resistance/susceptible response of maize to A. flavus. The miRNA hubs in resistance modules (TZAR102 and MI82) were mostly connected to transcription factors and protein kinases. Specifically, the module of miRNA zma-miR156b-nb – squamosa promoter binding protein (SBP), zma-miR398a-3p – SKIP5, and zma-miR394a-5p – F-box protein 6 combinations in the resistance-associated modules were considered important candidates for future functional studies. [ABSTRACT FROM AUTHOR]

Background: Aspergillus flavus is an important agricultural and food safety threat due to its production of carcinogenic aflatoxins. It has high level of genetic diversity that is adapted to various environments. Recently, we reported two reference genomes of A. flavus isolates, AF13 (MAT1-2 and highly aflatoxigenic isolate) and NRRL3357 (MAT1-1 and moderate aflatoxin producer). Where, an insertion of 310 kb in AF13 included an aflatoxin producing gene bZIP transcription factor, named atfC. Observations of significant genomic variants between these isolates of contrasting phenotypes prompted an investigation into variation among other agricultural isolates of A. flavus with the goal of discovering novel genes potentially associated with aflatoxin production regulation. Present study was designed with three main objectives: (1) collection of large number of A. flavus isolates from diverse sources including maize plants and field soils; (2) whole genome sequencing of collected isolates and development of a pangenome; and (3) pangenome-wide association study (Pan-GWAS) to identify novel secondary metabolite cluster genes. Results: Pangenome analysis of 346 A. flavus isolates identified a total of 17,855 unique orthologous gene clusters, with mere 41% (7,315) core genes and 59% (10,540) accessory genes indicating accumulation of high genomic diversity during domestication. 5,994 orthologous gene clusters in accessory genome not annotated in either the A. flavus AF13 or NRRL3357 reference genomes. Pan-genome wide association analysis of the genomic variations identified 391 significant associated pan-genes associated with aflatoxin production. Interestingly, most of the significantly associated pan-genes (94%; 369 associations) belonged to accessory genome indicating that genome expansion has resulted in the incorporation of new genes associated with aflatoxin and other secondary metabolites. Conclusion: In summary, this study provides complete pangenome framework for the species of Aspergillus flavus along with associated genes for pathogen survival and aflatoxin production. The large accessory genome indicated large genome diversity in the species A. flavus, however AflaPan is a closed pangenome represents optimum diversity of species A. flavus. Most importantly, the newly identified aflatoxin producing gene clusters will be a new source for seeking aflatoxin mitigation strategies and needs new attention in research. [ABSTRACT FROM AUTHOR]

Aims To develop antifungal lactic acid bacteria (LAB) and investigate their antifungal mechanisms against Aspergillus flavus in aflatoxin (AF) production. Methods and results We isolated 179 LABs from cereal-based fermentation starters and investigated their antifungal mechanism against A. flavus through liquid chromatography-mass spectrometry and co-culture analysis techniques. Of the 179 isolates, antifungal activity was identified in Pediococcus pentosaceus, Lactobacillus crustorum , and Weissella paramesenteroides. These LABs reduced AF concentration by (i) inhibiting mycelial growth, (ii) binding AF to the cell wall, and (iii) producing antifungal compounds. Species-specific activities were also observed, with P. pentosaceus inhibiting AF production and W. paramesenteroides showing AF B1 binding activity. In addition, crucial extracellular metabolites for selecting antifungal LAB were involved in the 2′,3′-cAMP-adenosine and nucleoside pathways. Conclusions This study demonstrates that P. pentosaceus, L. crustorum , and W. paramesenteroides are key LAB strains with distinct antifungal mechanisms against A. flavus , suggesting their potential as biological agents to reduce AF in food materials. [ABSTRACT FROM AUTHOR]

The fungal cell wall serves as the primary interface between fungi and their external environment, providing protection and facilitating interactions with the surroundings. Chitin is a vital structural element in fungal cell wall. Chitin deacetylase (CDA) can transform chitin into chitosan through deacetylation, providing various biological functions across fungal species. Although this modification is widespread in fungi, the biological functions of CDA enzymes in Aspergillus flavus remain largely unexplored. In this study, we aimed to investigate the biofunctions of the CDA family in A. flavus. The A. flavus genome contains six annotated putative chitin deacetylases. We constructed knockout strains targeting each member of the CDA family, including Δcda1, Δcda2, Δcda3, Δcda4, Δcda5, and Δcda6. Functional analyses revealed that the deletion of CDA family members neither significantly affects the chitin content nor exhibits the expected chitin deacetylation function in A. flavus. However, the Δcda6 strain displayed distinct phenotypic characteristics compared to the wild-type (WT), including an increased conidia count, decreased mycelium production, heightened aflatoxin production, and impaired seed colonization. Subcellular localization experiments indicated the cellular localization of CDA6 protein within the cell wall of A. flavus filaments. Moreover, our findings highlight the significance of the CBD1 and CBD2 structural domains in mediating the functional role of the CDA6 protein. Overall, we analyzed the gene functions of CDA family in A. flavus, which contribute to a deeper understanding of the mechanisms underlying aflatoxin contamination and lay the groundwork for potential biocontrol strategies targeting A. flavus. [ABSTRACT FROM AUTHOR]

Aflatoxin B1 (AFB1) and fumonisin B1 (FB1) are mycotoxins widely distributed in maize and maized-based products, often occurring together. The implications of co-exposure to aflatoxin and fumonsin for human health are numerous, but a particular concern is the potential of FB1 to modulate AFB1 hepatotoxicity. This study evaluated the toxicity of these mycotoxins, alone or combined, in a human non-tumorigenic liver cell line, HHL-16 cells, and assessed the effects of AFB1 and FB1 on expression of genes involved in immune and growth factor pathways. The results demonstrated that in HHL-16 cells, both AFB1 and FB1 had dose-dependent and time-dependent toxicity, and the combination of them showed a synergistic toxicity in the cells. Moreover, AFB1 caused upregulation of IL6 , CCL20 , and BMP2 , and downregulation of NDP. In combination of AFB1 with FB1, gene expression levels of IL6 and BMP2 were significantly higher compared to individual FB1 treatment, and had a tendency to be higher than individual AFB1 treatment. This study shows that FB1 may increase the hepatoxicity of AFB1 through increasing the inflammatory response and disrupting cell growth pathways. [ABSTRACT FROM AUTHOR]

The study was conducted to investigate the pathology of naturally occurring co-contamination of mycotoxins in poultry farms and the role of local management practices in the overall scenario. During the investigation, different chicken farms (n=73) in the Aizawl district of Mizoram state (India) were surveyed and monitored for management practices, the occurrence of disease and mortality. The feed samples collected from affected farms were tested for the presence of Aflatoxin and Ochratoxin by ELISA kits. The dead birds were subjected to necropsy. Liver and kidneys from the dead birds were processed for histopathological studies. Ten feed samples out of 49 tested (20.4%) were found affected by co-contamination with Aflatoxin and Ochratoxin. In this study, the mean level of Aflatoxin and Ochratoxin was 13.302 ppb and1.822 ppb, and the median level 12.143 ppb and1.674, ppb, respectively. Clinical signs were ruffled feathers, depression, dullness, huddling, poor growth and, anorexia. Necropsy revealed pathological lesions in visceral and lymphoid organs. Histopathological findings were inflammations, degenerative, and necrotic lesions in liver and kidneys. Poor management practices were observed in affected farms. A perusal of available literature did not reveal any study on the presence of co-contamination of Mycotoxicosis in poultry and poultry feed in Aizawl, Mizoram. [ABSTRACT FROM AUTHOR]

Childhood stunting is a global phenomenon affecting more than 149 million children under the age of 5 worldwide. Exposure to aflatoxins (AFs) in utero, during breastfeeding, and consumption of contaminated food affect the gut microbiome, resulting in intestinal dysfunction and potentially contributing to stunting. This review explores the potential relationship between AF exposure, environmental enteropathy and childhood stunting. AFs bind to DNA, disrupt protein synthesis and elicit environmental enteropathy (EE). An EE alters the structure of intestinal epithelial cells, impairs nutrient uptake and leads to malabsorption. This article proposes possible intervention strategies for researchers and policymakers to reduce AF exposure, EE and childhood stunting, such as exposure reduction, the implementation of good agricultural practices, dietary diversification and improving environmental water sanitation and hygiene. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Agriculture & Nature / Kahramanmaraş Sütçü İmam Üniversitesi Tarım & Doğa Dergisi is the property of Kahramanmaras Sutcu Imam Universitesi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Introduction: Identification of chemical toxins from complex or highly processed foods can present 'needle in the haystack' challenges for chemists. Metagenomic data can be used to guide chemical toxicity evaluations by providing DNA-based description of the wholistic composition (eukaryotic, bacterial, protozoal, viral, and antimicrobial resistance) of foods suspected to harbor toxins, allergens, or pathogens. This type of information can focus chemistry-based diagnostics, improve hazard characterization and risk assessment, and address data gaps. Additionally, there is increasing recognition that simultaneously co-occurring mycotoxins, either from single or multiple species, can impact dietary toxicity exposure. Metagenomic data provides a way to address data gaps related to cooccurrence of multiple fungal species. Methods: Paired metagenomic and chemical data were used to evaluate aflatoxin-contaminated kibble with known levels of specific mycotoxins. Kibble was ground to a fine powder for both chemical and molecular analyses. Chemical analyses were performed with Liquid Chromatography Mass Spectrometry (LCMS) and according to the AOAC Official method 2005.08: Aflatoxins in Corn, Raw Peanuts, and Peanut Butter using Liquid Chromatography with Post-Column Photochemical Derivatization. Metagenomes were created from DNA extracted from ground kibble and sequenced on an Illumina NextSeq 2000 with an average sequence depth of 180 million reads per replicate. Results and discussion: Metagenomic data demonstrated that the abundance of DNA from putative aflatoxigenic Aspergillus spp. correlated with the levels of aflatoxin quantified by LCMS. Metagenomic data also identified an expansive range of co-occurring fungal taxa which may produce additional mycotoxins. DNA data paired with chemical data provides a novel modality to address current data gaps surrounding dietary mycotoxin exposure, toxigenic fungal taxonomy, and mycotoxins of emerging concern. [ABSTRACT FROM AUTHOR]

Aspergillus flavus is a ubiquitous pathogen that can infect many foods and grains, and it produces large amounts of aflatoxins during their storage. Aflatoxins are considered highly toxic and carcinogenic to humans, and they cause great damage to crop production, food security, and human health. Thus, controlling A. flavus and aflatoxins in grains presents a great challenge to humans worldwide. Over the past decade, many strategies have been demonstrated to be useful in controlling A. flavus and aflatoxins during food storage. These methods involve physical agents, chemical agents, biological agents, etc. Some of these methods are currently used in actual production. In this review, we summarize the reported methods for controlling A. flavus and aflatoxins during food storage in the past ten years and elucidate their advantages and disadvantages. The methods discussed include irradiation technology; low oxygen atmospheres; chemical fungicides (benzalkonium chloride, iodine, ammonium bicarbonate, and phenolic and azole compounds); biological agents from plants, animals, and micro-organisms; and aflatoxin elimination methods. We expect that this review will promote the applications of current strategies and be useful for the development of novel technologies to prevent or eliminate A. flavus and aflatoxins in food and feed during storage. [ABSTRACT FROM AUTHOR]

This review explores the repercussions of mycotoxin contamination in food and feed, emphasising potential threats to agriculture, animal husbandry and public health. The primary objective is to make a comprehensive assessment of the neurotoxic consequences of mycotoxin exposure, an aspect less explored in current literature. Emphasis is placed on prominent mycotoxins, including aflatoxins, fumonisins, zearalenone (ZEA) and ochratoxins, known for inducing acute and chronic diseases such as liver damage, genetic mutation and cancer. To elucidate the effects, animal studies were conducted, revealing an association between mycotoxin exposure and neurological damage. This encompasses impairments in learning and memory, motor alterations, anxiety and depression. The underlying mechanisms involve oxidative stress, disrupting the balance between reactive oxygen species (ROS) and antioxidant capacity. This oxidative stress is linked to neuronal damage, brain inflammation, neurochemical imbalance, and subsequent behavioural changes. The review underscores the need for preventive measures against mycotoxin exposure. While complete avoidance is ideal, exploration into the potential use of antioxidants as a viable solution is discussed, given the widespread contamination of many food products. Specifically, the protective role of natural compounds, such as polyphenols, is highlighted, showcasing their efficacy in mitigating mycotoxicosis in the central nervous system (CNS), as evidenced by findings in various animal models. In summary, countering mycotoxin-induced neurotoxicity requires a multifaceted approach. The identified natural compounds show promise, but their practical use hinges on factors like bioavailability, toxicity and understanding their mechanisms of action. Extensive research is crucial, considering the diverse responses to different mycotoxins and neurological conditions. Successful implementation relies on factors such as the specific mycotoxin(s) involved and achievable effective concentrations. Further research and clinical trials are imperative to establish the safety and efficacy of these compounds in practical applications. [ABSTRACT FROM AUTHOR]

Simple Summary: Mycotoxins, which are secondary metabolites of fungi, have several adverse effects on both animal and human organisms. Because of climate change, mycotoxins are gradually spreading from the south to the north and west in Europe, causing a growing risk for feed and food. The toxic influences of mycotoxins are intensively studied in domestic animals, but much less is known about the game species. Reproductive abnormalities have been observed in a valuable game species, the fallow deer. We supposed mycotoxin contamination to be one of the possible causes because it is known to be dangerous even at low toxin levels, especially for young animals. As the mycotoxin exposure of the fallow deer living in a forest–agriculture complex habitat is not known, our goals were (i) to determine the mycotoxin presence in hinds; (ii) to detect the spatial and individual variability in mycotoxin levels; and (iii) to reveal the mycotoxin concentration in the fetuses in relation to their mothers. A total of 72 fallow deer embryos and their mothers were sampled in seven forested regions in Hungary in the 2020/2021 hunting season. Mycotoxin metabolites were analyzed by ELISA from the liver tissue. We detected the studied mycotoxins (Aflatoxin, Zearalenone, Fumonisin B1, DON, T2/HT2-toxin) in 41–96% of fallow deer hinds and 3–98% of the fetuses. All mycotoxins were passed into the fetus, but only Fumonisin B1 rarely passed. The individual variability of mycotoxin levels was extremely high, which obscured the spatial differences among the study sites. We concluded a possible accumulation of ZEA and DON in the fetuses because of the higher concentrations in the fetus than in the hind. These results reflect the potential threats of mycotoxins to the population dynamics and reproduction of wild fallow deer. Reproductive abnormalities have been observed in fallow deer populations in Hungary. We supposed mycotoxin contamination to be one of the possible causes because multi-mycotoxin contamination is known to be dangerous even at low toxin levels, especially for young animals. We investigated the spatial pattern of mycotoxin occurrences and the relationship between maternal and fetal mycotoxin levels. A total of 72 fallow deer embryos and their mothers were sampled in seven forested regions in Hungary in the 2020/2021 hunting season. We analyzed Aflatoxin (AF), Zearalenone (ZEA), Fumonizin B1 (FB1), DON, and T2-toxin concentrations in maternal and fetal livers by ELISA. AF was present in 70% and 82%, ZEA in 41% and 96%, DON in 90% and 98%, T2-toxin in 96% and 85%, and FB1 in 84% and 3% of hind and fetus livers, respectively. All mycotoxins passed into the fetus, but only Fumonizin B1 rarely passed. The individual variability of mycotoxin levels was extremely high, but the spatial differences were moderate. We could not prove a relation between the maternal and fetal mycotoxin concentrations, but we found an accumulation of ZEA and DON in the fetuses. These results reflect the possible threats of mycotoxins to the population dynamics and reproduction of wild fallow deer. [ABSTRACT FROM AUTHOR]

Velvet (VeA), a light-regulated protein that shuttles between the cytoplasm and the nucleus, serves as a key global regulator of secondary metabolism in various Aspergillus species and plays a pivotal role in controlling multiple developmental processes. The gene vepN was chosen for further investigation through CHIP-seq analysis due to significant alterations in its interaction with VeA under varying conditions. This gene (AFLA_006970) contains a Septin-type guanine nucleotide-binding (G) domain, which has not been previously reported in Aspergillus flavus (A. flavus). The functional role of vepN in A. flavus was elucidated through the creation of a gene knockout mutant and a gene overexpression strain using a well-established dual-crossover recombinational technique. A comparison between the wild type (WT) and the ΔvepN mutant revealed distinct differences in morphology, reproductive capacity, colonization efficiency, and aflatoxin production. The mutant displayed reduced growth rate; dispersion of conidial heads; impaired cell wall integrity; and decreased sclerotia formation, colonization capacity, and aflatoxin levels. Notably, ΔvepN exhibited complete growth inhibition under specific stress conditions, highlighting the essential role of vepN in A. flavus. This study provides evidence that vepN positively influences aflatoxin production, morphological development, and pathogenicity in A. flavus. [ABSTRACT FROM AUTHOR]

Aflatoxin contamination remains a major global food safety challenge, with serious implications on agriculture and public health. Kenya has frequently experienced outbreaks of acute and chronic aflatoxin poisoning over the years with adverse human health effects. Farmers respond to incidences of aflatoxin contamination using a myriad of control practices. This study evaluates the choice of aflatoxin contamination control practices among smallholder maize farmers in Kilifi County, an aflatoxin hotspot in the Coast region of Kenya. A Poisson regression model was employed on a sample of 270 farmers selected using a multistage sampling technique. The aflatoxin contamination control practices used by maize farmers in Kilifi County can broadly be categorized into pre-harvest practices (timely planting, pest control, use of improved seed varieties, and early harvesting) and postharvest practices (sorting, proper drying, and use of storage pesticides). The Poisson regression estimates reveal that aflatoxin awareness, access to extension services, wealth status, and gender were the most important drivers of farmers choice of aflatoxin contamination control practices in Kilifi County. Thus, policy initiatives that improve farmers awareness and access to information are recommended in improving maize farmers aflatoxin contamination knowledge and uptake of management practices. To be effective, such interventions must address gender and wealth differences. [ABSTRACT FROM AUTHOR]

Mycotoxins are toxic mycological products that when consumed, absorbed or inhaled cause sickness or even the death of humans. Therefore, the present study aimed to evaluate the contamination levels of mycotoxins (aflatoxins, AFB1, AFB2, AFG1, AFG2, and ochratoxin A, OTA) in selected medicinal herbs and shrubs using thin‐layer chromatography (TLC) and high‐performance liquid chromatography (HPLC). A total of 15 samples of medicinal herbs and shrubs were selected. Among them, four samples were aflatoxin contaminated while two samples were ochratoxin A contaminated. The highest level of aflatoxin was detected in Justicia adhathoda (4,704.94 ppb) through HPLC (153.4 ppb) and through TLC, while the lowest level of aflatoxin was detected in Pegnum harmala (205.1 ppb) through HPLC. Similarly, the highest level of OTA was detected in Dodonia viscosa (0.53 ppb) through HPLC (0.5 ppb) and through TLC, while the lowest level was detected in J. adhathoda (O.11 ppb) through HPLC (0.4 ppb) and through TLC. The OTA concentration was very low, being negligible and below permissible limits. The present study concludes that there is a potential risk for the consumption of herbal decoctions. Therefore, regular monitoring and proper management of mycotoxins, including aflatoxins and OTA, in herbal medicines are needed to ensure the safety of herbal drugs to protect consumers. [ABSTRACT FROM AUTHOR]

The potential of Raman hyperspectral imaging with a 785 nm excitation line laser was examined for the detection of aflatoxin contamination in corn kernels. Nine-hundred kernels were artificially inoculated in the laboratory, with 300 kernels each inoculated with AF13 (aflatoxigenic) fungus, AF36 (nonaflatoxigenic) fungus, and sterile distilled water (control). One-hundred kernels from each treatment were subsequently incubated for 3, 5, and 8 days. The mean spectra of single kernels were extracted from the endosperm side and the embryo area of the germ side, and local Raman peaks were identified based upon the calculated reference spectra of aflatoxin-negative and -positive categories separately. The principal component analysis-linear discriminant analysis models were established using different types of variable inputs including original full spectra, preprocessed full spectra, and identified local peaks over kernel endosperm-side, germ-side, and both sides. The results of the established discriminant models showed that the germ-side spectra performed better than the endosperm-side spectra. Based upon the 20 ppb-threshold, the best mean prediction accuracy of 82.6% was achieved for the aflatoxin-negative category using the original spectra in the combined form of both kernel sides, and the best mean prediction accuracy of 86.7% was obtained for the -positive category using the preprocessed germ-side spectra. Based upon the 100 ppb-threshold, the best mean prediction accuracies of 85.0% and 89.6% were achieved for the aflatoxin-negative and -positive categories separately, using the same type of variable inputs for the 20 ppb-threshold. In terms of overall prediction accuracy, the models established upon the original spectra in the combined form of both kernel sides achieved the best predictive performance, regardless of the threshold. The mean overall prediction accuracies of 81.8% and 84.5% were achieved with the 20 ppb- and 100 ppb-thresholds, respectively.

SUMMARY: The poor quality and safety of poultry feed in developing countries pose significant concerns as they negatively impact poultry performance and present potential health risks to human consumers. Aflatoxicosis, resulting from postharvest contamination of feed ingredients by certain mold species, severely affects birds' overall health and productivity. To address these challenges, various feed additives have become increasingly important in poultry nutrition. Phytogenics, natural plant-derived products, have gained popularity for their ability to mitigate aflatoxicosis by their rich composition of bioactive compounds that offer beneficial effects on physiological and immunological functions. In light of the growing significance of aflatoxicosis in poultry nutrition, this review provides an overview of aflatoxins and their harmful effects on poultry health. It highlights the role of phytogenic compounds as growth and health stimulators, emphasizing their potential application in poultry nutrition. Furthermore, the review explores the antioxidant and immunological mechanisms through which phytogenics mitigate aflatoxicosis in poultry, including specific compounds and pathways involved. The findings reveal that aflatoxins induce inflammation, and oxidative stress in poultry, resulting in cellular damage and immune dysfunction. Phytogenics play a crucial role in ameliorating the harmful effects of aflatoxins in birds by countering aflatoxin-induced oxidative stress, reducing inflammation, and restoring gut health and immune functions. In conclusion, phytogenics offer an invaluable tool for the mitigation of aflatoxicosis in poultry and serve as potent natural and safe alternatives to antibiotics and synthetic antioxidants, promoting sustainable production practices.