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Co-occurrence of multiple mycotoxins is a growing global food safety concern due to their harmful effects on humans and animals. This study developed an eco-friendly sample preparation method and an innovative multiplex microarray-based lateral flow immunoassay, using a novel portable reader for on-site simultaneous determination of five regulated mycotoxins—aflatoxin B1, T-2 toxin, zearalenone, deoxynivalenol, and fumonisin B1 in rice. The eco-friendly and ultrafast extraction procedure utilizes a bio-based solvent. Principally, the microarray signals generated through a novel luminescent organic dye were captured to quantify mycotoxin levels in samples using a portable reader installed with a user-friendly interface. The assay demonstrates accurate detection and quantification of these mycotoxins, with recoveries ranging from 77% to 127%. Detection limits ranged from 0.56 to 1.89 μg/kg, which were well below their regulatory limits, with a relative standard deviation below 25%. This analytical system provides an on-site method for detecting multiple mycotoxins in rice. [ABSTRACT FROM AUTHOR]

Gold-grafted melamine sponge (AuSp) was fabricated and applied as surface-enhanced Raman spectroscopy (SERS) substrate for enzyme-linked immunoassay of 2 typical mycotoxins, zearalenone (ZEN) and T-2 toxin. The gold nanoparticles were in situ grown on the sponge, and the product showed good effect in enhancing Raman signal and good stability. COMSOL and Gaussian simulations were used to analyze the enhancing effect. Some of the experimental conditions that may affect the determination results were studied in order to meet the maximum response to mycotoxins. At optimized conditions, the limits of detection of the method were 1 μg·kg−1 and 0.05 μg·kg−1 for ZEN and T-2 toxin, with linear ranges of 5–100 μg·kg−1 and 0.1–20 μg·kg−1, respectively, and relative standard deviations lower than 7.0%. The established method was used to determine ZEN and T-2 toxin in six kinds of cereal samples, the recoveries were 85–120%, and relative errors were less than 13.0%. These results indicated the accuracy of the established method. After 1-year storage, signal strength of AuSp for mycotoxins determination decreased by less than 10%, showing stability over a long time. Our work provided a new method for SERS determination of mycotoxins, which reduces the complexity of substrate preparation and the sample consumption compared with traditional methods, and thus has potential for on-line SERS determination. [ABSTRACT FROM AUTHOR]

The contamination of food and feed by mycotoxins, particularly zearalenone (ZEA) and deoxynivalenol (DON), is a global issue. Prenatal exposure to ZEA and DON can result in congenital cardiac malformations in fetuses. Addressing the prevention and mitigation of embryonic cardiotoxicity caused by these toxins is crucial. Citrus limonoid nomilin (NOM) is an extract known for its pathological properties in various diseases. This study investigated the potential mechanism of NOM in mitigating cardiotoxicity caused by ZEA and DON co-exposure in a zebrafish model. The findings indicated that NOM pretreatment alleviated cardiac developmental toxicity induced by ZEA and DON and normalized the expression of key genes involved in heart development, including gata4, vmhc, nkx2.5, and sox9b. Co-exposure to NOM, ZEA, and DON enhanced SOD and catalase activity, increased glutathione levels, and reduced ROS and malondialdehyde production. Furthermore, NOM reduced cardiac oxidative damage by activating the Keap1/Nrf2 signaling pathway. In summary, this study offers new insights for preventive interventions against congenital heart disease caused by mycotoxin exposure. [ABSTRACT FROM AUTHOR]

A QuEChERS (quick, easy, cheap, effective, rugged, and safe)-based multi-mycotoxin method was developed, analyzing 24 (17 free and 7 modified) Alternaria and Fusarium toxins in cereals via ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). A modified QuEChERS approach was optimized for sample preparation. Quantification was conducted using a combination of stable isotope dilution analysis (SIDA) for nine toxins and matrix-matched calibration for ten toxins. Quantification via a structurally similar internal standard was conducted for four analytes. Alternariol-9-sulfate (AOH-9-S) was measured qualitatively. Limits of detection (LODs) were between 0.004 µg/kg for enniatin A1 (ENN A1) and 3.16 µg/kg for nivalenol (NIV), while the limits of quantification were between 0.013 and 11.8 µg/kg, respectively. The method was successfully applied to analyze 136 cereals and cereal-based foods, including 28 cereal-based infant food products. The analyzed samples were frequently contaminated with Alternaria toxins, proving their ubiquitous occurrence. Interestingly, in many of those samples, some modified Alternaria toxins occurred, mainly alternariol-3-sulfate (AOH-3-S) and alternariol monomethyl ether-3-sulfate (AME-3-S), thus highlighting the importance of including modified mycotoxins in the routine analysis as they may significantly add to the total exposure of their parent toxins. Over 95% of the analyzed samples were contaminated with at least one toxin. Despite the general contamination, no maximum or indicative levels were exceeded. [ABSTRACT FROM AUTHOR]

The present study revealed the consequences of the interaction of a widely used bioinsecticide and endophyte Metarhizium anisopliae with the hazardous mycotoxin zearalenone (ZEN) as a pure substance and with ZEN as a native component of a crude Fusarium extract. In the environment, microorganisms encounter a mixture of metabolites secreted by other organisms living in the same area, not single substances. The obtained results suggest that M. anisopliae, exposed to a variety of active substances produced by Fusarium graminearum, is able to eliminate ZEN. Within 14 days, M. anisopliae biotransformed 90.8% and 85.8% of ZEN as a pure substance and ZEN as a native component of the F. graminearum extract from Rice Medium (E-Fg-RM), respectively, through reduction predominantly to α-epimers of zearalenols and zearalanols, considered more estrogenic than ZEN, which can raise concerns. Compared to pure ZEN, E-Fg-RM significantly affected the production of Metarhizium secondary metabolites by increasing the destruxins amount by approximately 20–25% and reducing the swainsonine content by 96.2%. All these findings provide a possible picture of the interaction of M. anisopliae with ZEN in the wild, mainly as a result of the use of crude extract from Fusarium, which contained a mixture of different metabolites. [ABSTRACT FROM AUTHOR]

Consumers are increasingly looking for healthier and sustainable diets. Plant-based diets rich in legumes satisfy this demand. Legumes contain protein, dietary fibers and starch. Technological processes can separate these fractions, which can be used as supplements, or as ingredients. Nonetheless, legumes are susceptible to fungal infection, causing a potential health concern, since some fungi can produce mycotoxins: toxic secondary metabolites. The aim of this work was to analyze the fate of mycotoxins during different stages of the production process of legume derived products from the raw materials to final products. An extraction followed by liquid chromatography-tandem mass spectrometry was used for the analysis, revealing the presence of enniatin B (ENN B), alternariol monomethyl ether (AME), deoxynivalenol, T2-toxin, nivalenol, fumonisin B1 and sterigmatocystin in raw materials, intermediate products and side streams. The alkaline solubilization steps, were effective in reducing ENN B; however, AME was found in one of the final products. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) is a fungal toxin produced by Fusarium that widely occurs in various types of grains and feed. It has strong reproductive toxicity and a wide range of distribution, which will impact food and feed safety. In this study, Bacillus subtilis ZENL09, which is capable of transforming ZEN was investigated. By optimizing the fermentation conditions, the supernatant of Bacillus subtilis ZENL09 fermentation can achieve about 88% degradation rate of ZEN after reacting with ZEN for 12 h. The degradation product was collected by high-performance liquid chromatography, followed by LC-Q-TOF and MS/MS analysis to identify the changes in molecular mass and compound structure. The possible structure of the degradation product was also tentatively determined. Further evaluation of the degradation products' cytotoxicity was conducted using HepG2 cells, and the results indicated a significant reduction in toxicity compared to ZEN. Detoxification experiments on corn steep liquor showed that the fermentation supernatant of Bacillus subtilis degraded 87% of ZEN within 12 h, which further validates its potential application for detoxification in food and agricultural products. [ABSTRACT FROM AUTHOR]

The present study aimed to find whether low doses of mixed mycotoxins would affect egg quality in laying hens, and to explore the oxidative stress induced liver damage through endoplasmic reticulum during summer stress. A total of 96 Jinghong laying hens, 36 wks of age, were divided into four treatments, with eight repetitions per treatment and three hens per repetition. All the hens were raised in summer (average temperature: 31.3 ± 0.5℃; average humidity: 85.5 ± 0.2%) for 28d. One treatment was fed a basal diet as control (CON), and the other three treatments were fed the same diets containing 3.0 mg/kg deoxynivalenol (DON), 0.5 mg/kg T-2 toxin (T-2), and 1.5 mg/kg DON + 0.25 mg/kg T-2 toxin (Mix). Albumen height and Haugh unit were decreased (P < 0.05) in the Mix group on day 14 and 28. The activity of total antioxidant capacity, glutathione peroxidase, catalase, and superoxide dismutase were decreased (P < 0.05) in the DON, T-2, and Mix groups. The alkaline phosphatase level in DON, T-2, and Mix groups was significantly increased (P < 0.05). The level of interleukin-1β, interferon-γ, and tumor necrosis factor-α in the Mix group were higher (P < 0.05) than CON, DON, and T-2 groups. Mix group upregulated the mRNA expressions of protein kinase RNA-like ER kinase, activating transcription factor4, IL-1β, nuclear factor-κ-gene binding, and nuclear respiratory factor 2 in the liver (P < 0.05). The results showed that low doses of DON and T-2 toxin could cause oxidative stress in the liver, but DON and T-2 toxin have a cumulative effect on virulence, which can reduce egg quality and cause endoplasmic reticulum stress in the liver. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is a mycotoxin commonly found in improperly stored grains, posing significant risks to food safety. Accurate detection of this mycotoxin in food matrices is crucial for ensuring consumer protection. In this study, we used in silico methods to design new peptides with high affinity and selectivity for recognizing OTA, starting crystal structures of OTA-protein complexes, which served to identify short key peptides within the interaction regions. Through deliberate point mutations, the stability and affinity of these peptides towards OTA were enhanced. To assess their suitability as capture probes for OTA sensing, we investigated the interaction of the selected peptides with OTA in a saline aqueous solution to mimic wet-lab experiments. Molecular docking, molecular dynamics simulations were used to evaluate the affinity and stability of the peptide-OTA complexes. Various parameters, including conformational changes, Molecular Mechanics Poisson–Boltzmann Surface Area energetics, the number of contacts, and the number of hydrogen bonds were analyzed to identify the most promising candidates. Among them, the peptide CWC11 has the highest affinity to OTA. Next, CWC11 peptide was used to functionalize the surfaces of two different carbonaceous nanomaterials: reduced graphene oxide (rGO) and graphene oxide (GO). The performance of the peptide-functionalized surfaces in presence of OTA was evaluated, revealing that GO exhibited superior performance compared to rGO. This study provides valuable insights into the design OTA-binding peptides and their use for the functionalization of carbonaceous surfaces, emphasizing the importance of surface selection to tune the efficiency and sensitivity of the detection. [ABSTRACT FROM AUTHOR]

Hippocampal neurons maintain the ability of proliferation throughout life to support neurogenesis. Deoxynivalenol (DON) is a mycotoxin that exhibits brain toxicity, yet whether and how DON affects hippocampal neurogenesis remains unknown. Here, we use mouse hippocampal neuron cells (HT-22) as a model to illustrate the effects of DON on neuron proliferation and to explore underlying mechanisms. DON exposure significantly inhibits the proliferation of HT-22 cells, which is associated with an up-regulation of cell cycle inhibitor p21 at both mRNA and protein levels. Global and site-specific m6A methylation levels on the 3'UTR of p21 mRNA are significantly increased in response to DON treatment, whereas inhibition of m6A hypermethylation significantly alleviates DON-induced cell cycle arrest. Further mechanistic studies indicate that the m6A readers YTHDF1 and IGF2BP1 are responsible for m6A-mediated increase in p21 mRNA stability. Meanwhile, 3'UTR of E3 ubiquitin ligase TRIM21 mRNA is also m6A hypermethylated, and another m6A reader YTHDF2 binds to the m6A sites, leading to decreased TRIM21 mRNA stability. Consequently, TRIM21 suppression impairs ubiquitin-mediated p21 protein degradation. Taken together, m6A-mediated upregulation of p21, at both post-transcriptional and post-translational levels, contributes to DON-induced inhibition of hippocampal neuron proliferation. These results may provide new insights for epigenetic therapy of neurodegenerative diseases. DON inhibits the proliferation of HT-22 cells. RNA m6A hypermethylation on the transcript of p21 enhances the mRNA stability in a YTHDF1- and IGF2BP1-dependent manner, which leads to the upregulation of p21. RNA m6A hypermethylation on the transcript of TRIM21 decreases the mRNA stability in a YTHDF2-dependent manner, which contributes to prevent p21 ubiquitin-mediated degradation. High expression of p21 contributes to inhibit cell proliferation. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN), produced by Fusarium species, is a potential risk to human health. Traditional enzyme-linked immunosorbent assay (ELISA) is restricted due to low sensitivity for the detection of ZEN. Herein, enzyme nanocomposites (ALP-SA-Bio-ssDNA, ASBD) were prepared with the self-assembly strategy based on streptavidin-labeled alkaline phosphatase (SA-ALP) and dual-biotinylated ssDNA (B2-ssDNA). The enzyme nanocomposites improved the loading amount of ALP and catalyzed more ascorbic acid 2-phosphate to generate ascorbic acid (AA). Subsequently, Cu2+ could be reduced to copper nanoclusters (CuNCs) having strong fluorescence signal by AA with poly T. Benefiting from the high enzyme load of nanocomposites and the strong signal of CuNCs, the fluorescence ELISA was successfully established for the detection of ZEN. The proposed method exhibited lower limit of detection (0.26 ng mL−1) than traditional ELISA (1.55 ng mL−1). The recovery rates ranged from 92.00% to 108.38% (coefficient of variation < 9.50%) for the detection of zearalenone in corn and wheat samples. In addition, the proposed method exhibited no cross reaction with four other mycotoxins. This proposed method could be used in trace detection for food safety. [ABSTRACT FROM AUTHOR]

Cattle are deemed less susceptible to mycotoxins due to the limited internal exposure resulting from rumen microbiota activity. However, the significant amounts of Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) frequently detected in bovine follicular fluid samples suggest that they could affect ovarian function. Both mycotoxins trigger several patterns of cell death and activate the NLRP3 inflammasome in the intestine. In vitro studies have reported a number of adverse effects on bovine oocytes. However, the biological relevance of such findings with regard to realistic concentrations of DON and ZEN in bovine follicular fluid is still not clear. Hence, it is important to better characterize the effects of dietary exposure to DON and ZEN on the bovine ovary. Using bovine primary theca cells, this study investigated the effects of real-life patterns for bovine ovary exposure to DON and ZEN, but also DON metabolite DOM-1, on cell death and NLRP3 inflammasome activation. Exposure to DON starting from 0.1 μM significantly decreased theca cell viability. The kinetics of phosphatidylserine translocation and loss of membrane integrity showed that ZEN and DON, but not DOM-1, induce an apoptotic phenotype. qPCR analysis of the expression of NLRP3, PYCARD, IL-1β, IL-18, and GSDMD in primary theca cells at concentrations of mycotoxin previously reported in cow follicular fluid clearly indicated that DON and DOM-1 individually and in mixture, but not ZEN, activate NLRP3 inflammasome. Altogether, these results suggest that real-life dietary exposure of cattle to DON may induce inflammatory disorders in the ovary. [ABSTRACT FROM AUTHOR]

Root lesion nematodes (RLN) of the genus Pratylenchus are causing significant damage in cereal production worldwide. Due to climate change and without efficient and environment-friendly treatments, the damages through RLNs are predicted to increase. Microscopic assessments of RLNs in the field and the greenhouses are time-consuming and laborious. As a result, cereal breeders have mostly ignored this pest. We present a method measuring RLN in infected cereal roots using a standardized PCR approach. Publicly available Pratylenchus neglectus primer combinations were evaluated. An optimal primer combination for RT-qPCR assay was identified to detect and quantify P. neglectus within infected cereal roots. Using the RT-qPCR detection assay, P. neglectus could be clearly distinguished from other plant parasitic nematodes. We could identify P. neglectus DNA in barley and wheat roots as low as 0.863 and 0.916 ng/µl of total DNA, respectively. A single P. neglectus individual was detected in water suspension and within barley and wheat roots. The RT-qPCR detection assay provides a robust and accurate alternative to microscopic nematode identification and quantification. It could be of interest for resistance breeding, where large populations must be screened to detect and quantify P. neglectus in farmer's fields. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) and zearalenone (ZEN) are often detected in plant materials used to produce feed for pre-pubertal gilts. Daily exposure to small amounts of these mycotoxins causes subclinical conditions in pigs and affects various biological processes (e.g. mycotoxin biotransformation). The aim of this preclinical study was to evaluate the effect of low monotonic doses of DON and ZEN (12 µg/kg body weight—BW—and 40 µg/kg BW, respectively), administered alone or in combination to 36 prepubertal gilts for 42 days, on the degree of immunohistochemical expression of oestrogen receptors (ERs) in the liver and the mRNA expression of genes encoding selected liver enzymes during biotransformation processes. The level of expression of the analysed genes proves that the tested mycotoxins exhibit variable biological activity at different stages of biotransformation. The biological activity of low doses of mycotoxins determines their metabolic activity. Therefore, taking into account the impact of low doses of mycotoxins on energy-intensive processes and their endogenous metabolism, it seems that the observed situation may lead to the activation of adaptation mechanisms. [ABSTRACT FROM AUTHOR]

To assess the impact of black-colored grain on Alternaria mycotoxin concentrations in different stages of the brewing process, brewing experiments were conducted in a microscale brewhouse. Different mixtures of visually unaffected and black-colored batches of two malt samples were used, which were obtained by an optical sorting device. The 13 Alternaria mycotoxins alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA), tentoxin (TEN), alterperylenol (ALTP), altertoxins I and II (ATX I and II), altenuene (ALT) as well as the modified forms AOH-3-glucoside (AOH-3-G), AOH-9-glucoside (AOH-9-G), AME-3-gluoside (AME-3-G), AOH-3-sulfate (AOH-3-S) and AME-3-sulfate (AME-3-S) were analyzed in each processing step by liquid chromatography–tandem mass spectrometry (LC–MS/MS), and the toxin concentrations were balanced over the whole brewing process. Fungal DNA content in the starting material (mixtures) was determined by quantitative real-time polymerase chain reaction (qPCR). In this study, TeA was the only toxin to migrate into the final beer, while the AOH, AME, TEN, ALTP and ATX I toxins were mainly found in the spent grains. The observance of AOH-3-S and AME-3-S in some processing steps also showed the possibility of modification reactions during brewing. Furthermore, no distinct correlations between the fungal DNA and the analyzed mycotoxins could be observed in the starting material, while the amount of black colored grains only impacted toxin concentrations in one of the two used malt samples. Nevertheless, it was shown that optical sorting of malt batches might be a useful tool for the malting and brewing industry to prevent elevated mycotoxin concentrations. [ABSTRACT FROM AUTHOR]

Zearalenone, an estrogenic mycotoxin, is widely exists in rice and agricultural products. Zearalenone has side effects on animal's reproduction and immunologic functions. Moreover, it can stimulate the proliferation of MCF-7 cells through estrogenic effects. The aim of this research is to understand the interactive effect of zearalenone and bioactive components of Hashemi brown and white rice extracts and γ-oryzanol on proliferation of MCF-7 cell, which were determined through Resazurin assay, ROS generation and flow cytometry. Results showed that zearalenone (10− 4 to 1 µM) could stimulate the proliferation of MCF-7 cells. The results showed that the various fractions of Hashemi brown (methanol, ethyl acetate and butanol) extracts, white rice (methanol, dichloromethane and ethylacetate) extracts and γ-Oryzanol could significantly decrease the estrogenic effects and as a consequence inhibit the proliferation of MCF-7 cells. So that, methanol, butanol and ethylacetate fractions of brown rice and γ-oryzanol significantly alleviate the cell proliferative effect of zearalenone (0.01 µM) from 1.17 ± 0.0063 to 0.97 ± 0.0064, 0.97 ± 0.006, 1.01 ± 0.004 and 0.97 ± 0.009, respectively. γ-Oryzanol and hashemi brown and white rice could also decrease the ROS generation and apoptosis rate in MCF-7 cells induced by zearalenone. Thus, it seems the estrogenic and cytotoxic effect of zearalenone may be modulated when the whole rice is used and rice extract protect against the harm of zearalenone. [ABSTRACT FROM AUTHOR]

The efficiency, transformation products, and mechanism of phycocyanin removal from water by simulated sunlight/Cu-decorated TiO2 photocatalyst treatment were studied. After 360 min of photocatalytic degradation, the removal rate of PC was higher than 96%, about 47% of DON was oxidized into NH4+-N, NO3− and NO2−. ·OH was the main active species in the photocatalytic system, which contributes about 55.7% to PC degradation efficiency, H+ and ·O2− also contributed to the photocatalytic activity. The degradation process of phycocyanin is firstly caused by the attack of free radicals, which leads to the disintegration of the chromophore group PCB and the apoprotein, and then apoprotein peptide chain was broken to generate small molecule dipeptides, amino acids, and their derivatives. Amino acid residues sensitive to free radical action in phycocyanin peptide chain include most hydrophobic amino acids such as leucine, isoleucine, proline, valine, phenylalanine, and some hydrophilic amino acids which are easily oxidized such as lysine and arginine. Small molecular peptides (dipeptides), amino acids, and their derivatives are broken off and released into water bodies for further reaction and degradation into smaller molecular weight substances. During this process, part of organic nitrogen was transferred to inorganic nitrogen. When photocatalytic oxidation lasts for 300 min, NH4+ increases from 0.41 mg/L to 2.21 mg/L, and DON removal rate reaches 47%. The Cu-TiO2 photocatalyst was found to decrease the CHCl3 formation potential; however, it exacerbated the production of dichloroacetamide (DCAcAm) and dichloroacetonitrile (DCAN) beyond their initial levels. The divergent trends of these disinfection by-products are due to the fundamental differences in the precursor material. [ABSTRACT FROM AUTHOR]

T-2 toxin is a worldwide problem for feed and food safety, leading to livestock and human health risks. The objective of this study was to explore the mechanism of T-2 toxin-induced small intestine injury in broilers by integrating the advanced microbiomic, metabolomic and transcriptomic technologies. Four groups of 1-day-old male broilers (n = 4 cages/group, 6 birds/cage) were fed a control diet and control diet supplemented with T-2 toxin at 1.0, 3.0, and 6.0 mg/kg, respectively, for 2 weeks. Compared with the control, dietary T-2 toxin reduced feed intake, body weight gain, feed conversion ratio, and the apparent metabolic rates and induced histopathological lesions in the small intestine to varying degrees by different doses. Furthermore, the T-2 toxin decreased the activities of glutathione peroxidase, thioredoxin reductase and total antioxidant capacity but increased the concentrations of protein carbonyl and malondialdehyde in the duodenum in a dose-dependent manner. Moreover, the integrated microbiomic, metabolomic and transcriptomic analysis results revealed that the microbes, metabolites, and transcripts were primarily involved in the regulation of nucleotide and glycerophospholipid metabolism, redox homeostasis, inflammation, and apoptosis were related to the T-2 toxin-induced intestinal damage. In summary, the present study systematically elucidated the intestinal toxic mechanisms of T-2 toxin, which provides novel ideas to develop a detoxification strategy for T-2 toxin in animals. [ABSTRACT FROM AUTHOR]

Alimentary toxic aleukia (ATA) is correlated with consuming grains contaminated by Fusarium species, particularly T-2 toxin, which causes serious hurt to human and animal health, chiefly in disorders of the haematopoietic system. However, the mechanism of haematopoietic dysfunction induced by T-2 toxin and the possible target pathway for the treatment of T-2 toxin-induced haematopoietic disorder of ATA remains unclear. In this study, genomes and proteomics were used for the first time to investigate the key differential genes and proteins that inhibit erythroid differentiation of K562 cells caused by T-2 toxin, and it was found that heat shock protein 27 (HSP27) and membrane-spanning 4-domains, subfamily A, member 3 (MS4A3) may play an important role in erythroid differentiation. Meanwhile, MS4A3 interference can inhibit the occurrence of erythroid differentiation of K562 cells and promote the phosphorylation of HSP27. Moreover, the binding of HSP27 to MS4A3 in natural state can activate the phosphorylation site of HSP27 (Ser-83), while T-2 toxin can abolish the activation of phosphorylation site by inhibiting the expression of MS4A3. These findings for the first time demonstrated that the MS4A3-HSP27 pathway may function an efficient therapeutic target pathway for treating T-2 toxin elicited haematopoietic disorders of ATA. [ABSTRACT FROM AUTHOR]

A nanozyme sensor array based on the ssDNA-distensible C3N4 nanosheet sensor elements for discriminating multiple mycotoxins commonly existing in contaminated cereals has been explored. The sensor array exploited (a) three DNA nonspecific sequences (A40, T40, C40) absorbed on the C3N4 nanosheets as sensor elements catalyzing the oxidation of TMB; (b) the presence of five mycotoxins affected the catalytic activity of three nanozymes with various degrees. The parameter (A0-A) was employed as the signal output to obtain the response patterns for different mycotoxins with the same concentration where A0 and A were the absorption peak values at 650 nm of oxTMB in the absence and presence of target mycotoxins, respectively. After the raw data was subjected to principal component analysis, 3D canonical score plots were obtained. The sensor array was capable of separating five mycotoxins from each other with 100% accuracy even if the concentration of the mycotoxins was as low as 1 nM. Moreover, the array performed well in discriminating the mycotoxin mixtures with different ratios. Importantly, the practicality of this sensor array was demonstrated by discriminating the five mycotoxins spiking in corn-free samples in 3D canonical score plots, validating that the sensor array can act as a flexible detection tool for food safety. A nanozyme sensor array was developed based on the ssDNA-distensible C3N4 NSs sensor elements for discriminating muitiple mycotoxins. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) and its derivatives are one of the most contaminated fungal toxins worldwide, posing a severe threat to food security and human life. Traditional physical and chemical detoxifying methods are unsatisfactory due to incomplete detoxification, nutrient loss, and secondary pollutants. In recent years, bioremediation for eliminating fungal toxins has been gradually investigated. ZEN lactone hydrolase (lactonase) has been widely studied because of its high activity, mild conditions, and non-toxic product property. This review comprehensively represents the gene mining, characterization, molecular modification, and application of microbial-derived ZEN lactonases. It is aimed to elucidate the advantages and challenges of ZEN lactonases in industrial application, which also provides perspectives on obtaining innovative and promising biocatalysts for ZEN degradation. Key points: • A timely and concise review related to enzymatic elimination towards ZEN is shown. • The catalytic conditions and mechanism of ZEN lactonase is presented. • The modification and application of ZEN lactonase are exhibited also. [ABSTRACT FROM AUTHOR]

Infants are sensitive to negative effects caused by food contaminants such as mycotoxins. To date, analytical methods assessing mycotoxin mixture exposure in infant stool are absent. Herein, we present a novel multi-mycotoxin LC-MS/MS assay capable of detecting 30+ analytes including the regulated mycotoxin classes (aflatoxins, trichothecenes, ochratoxins, zearalenone, citrinin), emerging Alternaria and Fusarium toxins, and several key metabolites. Sample preparation consisted of a 'dilute, filter, and shoot' approach. The method was in-house validated and demonstrated that 25 analytes fulfilled all required criteria despite the high diversity of chemical structures included. Extraction recoveries for most of the analytes were in the range of 65–114% with standard deviations below 30% and limits of detection between 0.03 and 11.3 ng/g dry weight. To prove the methods' applicability, 22 human stool samples from premature Austrian infants (n = 12) and 12-month-old Nigerian infants (n = 10) were analyzed. The majority of the Nigerian samples were contaminated with alternariol monomethyl ether (8/10) and fumonisin B1 (8/10), while fumonisin B2 and citrinin were quantified in some samples. No mycotoxins were detected in any of the Austrian samples. The method can be used for sensitive human biomonitoring (HBM) purposes and to support exposure and, potentially, risk assessment of mycotoxins. Moreover, it allows for investigating potential associations between toxicant exposure and the infants' developing gut microbiome. [ABSTRACT FROM AUTHOR]

Mycotoxin contamination of food and feed is a serious food safety issue and causes acute and chronic diseases in humans and livestock. Climatic and agronomic changes helps in the proliferation of fungal growth and mycotoxin production in food commodities. Mycotoxin contamination has attracted global attention due to its wide range of toxicity to humans and animals. However, physical and chemical management approaches in practice are unsafe for well-being due to their health-hazardous nature. Various antibiotics and preservatives are in use to reduce the microbial load and improve the shelf life of food products. In addition, the use of antibiotic growth promotors in livestock production may increase the risk of antimicrobial resistance, which is a global health concern. Due to their many uses, probiotics are helpful microbes that have a significant impact on food and nutrition. Furthermore, the probiotic potential of lactic acid bacteria (LAB) is employed in various food and feed preparations to neutralize mycotoxins, antimicrobial activities, balance the gut microbiome, and various immunomodulatory activities in both humans and livestock. In addition, LAB produces various antimicrobials, flavouring agents, peptides, and proteins linked to various food and health care applications. The LAB-based processes for mycotoxin management are more effective, eco-friendly, and low-cost than physical and chemical approaches. The toxicity, novel preventive measures, binding nature, and molecular mechanisms of mycotoxins' detoxification using LAB have been highlighted in this review. [ABSTRACT FROM AUTHOR]

Mycotoxins are low molecular weight toxic compounds, which can cause severe health problems in animals and humans. Immunoassays allow rapid, simple and cost-effective screening of mycotoxins. Sandwich assays with a direct readout provide great improvement in terms of selectivity and sensitivity, compared to the widely used competitive assay formats, for the analysis of low molecular weight molecules. In this work, we report a non-competitive fluorescence anti-immune complex (IC) immunoassay, based on the specific recognition of HT-2 toxin with a pair of recombinant antibody fragments, namely antigen-binding fragment (Fab) (anti-HT-2 (10) Fab) and single-chain variable fragment (scFv) (anti-IC HT-2 (10) scFv). The SpyTag and SpyCatcher glue proteins were applied for the first time as a bioconjugation tool for the analysis of mycotoxins. To this aim, a SpyTag-mScarlet-I (fluorescent protein) and scFv-SpyCatcher fusion proteins were constructed, produced and fused in situ during the assay by spontaneous Tag-Catcher binding. The assay showed an excellent sensitivity with an EC50 of 4.8 ± 0.4 ng mL−1 and a dynamic range from 1.7 ± 0.3 to 13 ± 2 ng mL−1, an inter-day reproducibility of 8.5% and a high selectivity towards HT-2 toxin without cross-reactivity with other Fusarium toxins. The bioassay was applied to the analysis of the toxin in an oat reference material and in oat samples, with a LOD of 0.6 µg kg−1, and the results were validated by analysing a certificate reference material and by HPLC–MS/MS. [ABSTRACT FROM AUTHOR]

Phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway is one of the most deregulated signaling pathway in prostate cancer. It controls basic processes in cells: cell proliferation and death. Any disturbances in the balance between cell death and survival might result in carcinogenesis. Deoxynivalenol (DON) is one of the most common mycotoxins, a toxic metabolites of fungi, present in our everyday diet and feed. Although previous studies reported DON to induce oxidative stress, modulate steroidogenesis, DNA damage and cell cycle modulation triggering together its toxicity, its effect on normal prostate epithelial cells is not known. The aim of the study was to evaluate the effect of DON on the apoptosis and autophagy in normal prostate epithelial cells via modulation of PI3K/Akt signaling pathway. The results showed that DON in a dose of 30 µM and 10 µM induces oxidative stress, DNA damage and cell cycle arrest in G2/M cell cycle phase. The higher concentration of DON induces apoptosis, whereas lower one autophagy in PNT1A cells, indicating that modulation of PI3K/Akt by DON results in the induction of autophagy triggering apoptosis in normal prostate epithelial cells. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEA) is a mycotoxin with estrogenic activity whose main effect is to impair the reproductive systems of animals. It leads to reproductive disorders in livestock and thus causes serious losses to agriculture and animal husbandry. This study aims to examine whether ZEA induces toxicity in Leydig cells through the PI3K/AKT signaling pathway and also to investigate the role played by the upstream phosphatase and tensin homolog (PTEN) gene. An adenovirus vector model was constructed to interfere with the PTEN gene to investigate whether ZEA promotes the apoptosis of TM3 cells through the PI3K/AKT pathway. Apoptosis was detected cytometrically and the protein expression levels of PTEN, AKT, p-AKT, Bax, and Bcl-2 were evaluated via western blot analysis. The results show that ZEA induces apoptosis of TM3 cells. PTEN expression is significantly increased (P < 0.01), Bax expression is increased (P < 0.05), AKT and p-AKT expression of anti-apoptotic protein is significantly decreased (P < 0.01), and Bcl-2 protein expression is decreased (P < 0.05) in the ZEA group compared with the control group. In the shRNA+ZEA group, the expression levels of PTEN and Bax proteins are significantly decreased (P < 0.01), AKT protein is significantly increased (P < 0.01), and p-AKT protein is increased (P < 0.05) compared with the ZEA group. This study thus demonstrates that ZEA promotes apoptosis of TM3 cells by targeting PTEN and thus inhibiting the PI3K/AKT signal pathway. [ABSTRACT FROM AUTHOR]

The influence of previous crops (oat, maize, or rape) and six variety of hulled oats (Avena sativa L.) were assessed qualitatively and quantitatively as to yield, thousand grain weight, and grain size in observations from 2009 to 2012. Harvested grain samples were analysed for the presence of mycotoxins (T-2 and HT-2 toxins, deoxynivalenol, and nivalenol) using LC/MS–MS. Fusarium poae was the species most often found on oats. Flamingsprofi and Salo were the most sensitive varieties of oat. The sensitivity of the variety to lodging as well as the earliness of the variety was correlated with the occurrence of fusariosis on oats. The highest average occurrence of Fusaria on oat grains was after use of maize as the pre-crop. The oat samples were most often contaminated with nivalenol. Nivalenol positively correlated with the weight of a thousand grains of oat and negatively correlated with the oats grain falling through the sieve. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEA) and Deoxynivalenol (DON) are two mycotoxins highly detected in agricultural products and feed. Both mycotoxins produce reproductive toxicity and pose a serious threat to human and animal health, among which pigs are the most sensitive animals. Sertoli cells (SCs) play an important role in spermatogenesis; however, the combined toxicity of ZEA and DON and the screening of effective protective agents remains to be determined. By studying the effects of N-acetylcysteine (NAC) on the cells exposed to 20 μM of ZEA and 0.6 μM of DON, we explored the protective mechanism of NAC (4 mM) on the cytotoxic injury of piglets SCs induced by both mycotoxins. The results showed that the combination of ZEA and DON destroy organelles and SCs structures, NAC significantly alleviates the damage caused by ZEA and DON. NAC also significantly increased the expression and distribution of zonula occludens 1 (ZO-1), decreased the relative mRNA and protein expression levels of Bax, Bid, caspase-3, and caspase-9, and increased Bcl-2 expression level and inhibited the decrease of mitochondrial membrane potential. Further, NAC also eases the cell cycle arrest and oxidative stress caused by ZEA and DON. In summary, our results show that NAC could alleviate SCs injury via reducing the oxidative damage and apoptosis caused by ZEA and DON. [ABSTRACT FROM AUTHOR]

A new Trichoderma asperellum strain, TasT1, was isolated from the rhizosphere of Malus sieversii in Xinjiang Region of China. Its antifungal ability was evaluated by controlling root rot Fusarium oxysporum (Fox68) of M. sieversii. The results showed that TasT1 inhibited the growth of Fox68, with an inhibition rate of 66.7%. After treating Arabidopsis thaliana seedlings with different concentrations of TasT1 fermentation liquor, the root hair density of the seedlings changed significantly. Treatment with 60% fermentation liquor caused the root length to increase by 21%, and the fresh weight by 80%. Furthermore, TasT1 fermentation liquor improved the detoxification ability of A. thaliana seedlings against Fox68 toxin and greatly alleviated leaf wilting. Defense-related enzymes (e.g., peroxidase, POD) activity and malondialdehyde content of A. thaliana increased significantly. In addition, in the TasT1 + Fox68 group, the expression levels of PDF1.2B, PDF1.2C, and PR1 were upregulated by 5.2, 5.1, and 2.2 times respectively after treatment for 3 d. Meanwhile, we used GC–MS analysis of chemical component of TasT1 fermentation liquor and found that TasT1 fermentation liquor include 49 kinds of ingredients which contain a lot of antibacterial substances. Our findings suggest that TasT1 fermentation liquor could induce the accumulation of defense-related enzymes to improve disease resistance and growth of A. thaliana seedlings. TasT1 has the potential to control F. oxysporum (Fox68) and protect M. sieversii. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) is a mycotoxin that contaminates crops worldwide and whose toxic adverse effects are well documented. This study aims to evaluate the protective effect of gallic acid (GA) against biochemical, oxidative, inflammatory, and pathological changes in ZEN treated rats' hepatorenal system. Wistar rats (n = 50; 150 ± 30 g) were randomly grouped into five cohorts (= 10) specifically: Control (rat chow); ZEN alone (100 µg/kg; per os), GA alone (40 mg/kg; per os), ZEN + GA1 (100 µg/kg + 20 mg/kg per os) and ZEN + GA2 (100 µg/kg + 40 mg/kg per os) and the study was for 28 successive days. Upon terminal sacrifice, biomarkers of hepatorenal function and oxidative stress were analyzed. An assessment of cytokine levels (IL-1β, IL-10) and histopathology of the liver and kidneys was also performed. Relative to the control, serum levels of urea, creatinine, and hepatic transaminases increased significantly (p < 0.05) in the ZEN alone group and reduced in groups co-treated with GA. ZEN treatment further resulted in decreases in the rat's antioxidant status. The increase in the reactive oxygen and nitrogen species (RONS) and lipid peroxidation (LPO) levels caused by ZEN exposure was reduced by GA in a dose-dependent manner (p < 0.05). Furthermore, ZEN-mediated increase in nitric oxide (NO), xanthine oxidase (XO), IL-1β, and myeloperoxidase (MPO) levels and suppression of IL-10 levels were reversed in the liver and kidney of GA co-treated rats. The extent of ZEN-mediated hepatorenal lesions was reduced in rats co-treated with GA. Our findings suggest that GA effectively abated biochemical, oxido-inflammatory and histological alteration caused by ZEN exposure, limiting ZEN toxicity and cellular damage in rats' hepatic and renal tissues. [ABSTRACT FROM AUTHOR]

Dwarf bunt caused by the pathogen Tilletia controversa Kühn is one of the most serious quarantine diseases of winter wheat. Metabolomics studies provide detailed information about the biochemical changes at the cell and tissue levels of plants. In the present study, a liquid chromatography/mass spectrometry (LC/MS) metabolomics approach was used to investigate the changes in the grain metabolomics of infected and noninfected with T. controversa samples. PCA suggested that T. controversa-infected and noninfected samples were separated during the interaction. LC/MS analysis showed that 62 different metabolites were recorded in the grains, among which a total of 34 metabolites were upregulated and 28 metabolites were downregulated. Prostaglandins (PGs) and 9-hydroxyoctadecadienoic acids (9-HODEs) are fungal toxin-related substances, and their expression significantly increased in T. controversa-infected grains. Additionally, the concentrations of cucurbic acid and octadecatrienoic acid changed significantly after pathogen infection, which play a large role in plant defense. The eight different metabolic pathways activated during T. controversa and wheat plant interactions included phenylalanine metabolism, isoquinoline alkaloid biosynthesis, starch and sucrose metabolism, tyrosine metabolism, sphingolipid metabolism, arginine and proline metabolism, alanine, aspartate, and glutamate metabolism, and tryptophan metabolism. In conclusion, we found differences in the metabolic profiles of wheat grains after T. controversa infection. To our knowledge, this is the first study to evaluate the metabolites in wheat grains after T. controversa infection. [ABSTRACT FROM AUTHOR]

Plastic and straw coverage (PC and SC) are often combined with fungicide application but their influence on fungicide entry into soil and the resulting consequences for soil quality are still unknown. The objective of this study was to investigate the impact of PC and SC, combined with fungicide application, on soil residual concentrations of fungicides (fenhexamid, cyprodinil, and fludioxonil), soil fungal biomass, mycotoxin occurrence, and soil organic matter (SOM) decomposition, depending on soil depth (0–10, 10–30, 30–60 cm) and time (1 month prior to fungicide application and respectively 1 week, 5 weeks, and 4 months afterwards). Soil analyses comprised fungicides, fusarium mycotoxins (deoxynivalenol, 15-acetyldeoxynivalenol, nivalenol, and zearalenone), ergosterol, soil microbial carbon and nitrogen, soil organic carbon, dissolved organic carbon, and pH. Fludioxonil and cyprodinil concentrations were higher under SC than under PC 1 week and 5 weeks after fungicide application (up to three times in the topsoil) but no differences were observed anymore after 4 months. Fenhexamid was not detected, presumably because of its fast dissipation in soil. The higher fludioxonil and cyprodinil concentrations under SC strongly reduced the fungal biomass and shifted microbial community towards larger bacterial fraction in the topsoil and enhanced the abundance and concentration of deoxynivalenol and 15-acetyldeoxynivalenol 5 weeks after fungicide application. Independent from the different fungicide concentrations, the decomposition of SOM was temporarily reduced after fungicide application under both coverage types. However, although PC and SC caused different concentrations of fungicide residues in soil, their impact on the investigated soil parameters was minor and transient (< 4 months) and hence not critical for soil quality. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (vomitoxin, DON) is a secondary metabolite produced by Fusarium spp. fungi and it is one of the most prevalent mycotoxins worldwide. Crop infestation results not only in food and feed contamination, but also in direct dermal exposure, especially during harvest and food processing. To investigate the potential dermotoxicity of DON, epidermoid squamous cell carcinoma cells A431 were compared to primary human neonatal keratinocytes (HEKn) cells via proteome/phosphoproteome profiling. In A431 cells, 10 µM DON significantly down-regulated ribosomal proteins, as well as mitochondrial respiratory chain elements (OXPHOS regulation) and transport proteins (TOMM22; TOMM40; TOMM70A). Mitochondrial impairment was reflected in altered metabolic competence, apparently combined with interference of the lipid biosynthesis machinery. Functional effects on the cell membrane were confirmed by live cell imaging and membrane fluidity assays (0.1–10 µM DON). Moreover, a common denominator for both A431 and HEKn cells was a significant downregulation of the squalene synthase (FDFT1). In sum, proteome alterations could be traced back to the transcription factor Klf4, a crucial regulator of skin barrier function. Overall, these results describe decisive molecular events sustaining the capability of DON to impair skin barrier function. Proteome data generated in the study are fully accessible via ProteomeXchange with the accession numbers PXD011474 and PXD013613. [ABSTRACT FROM AUTHOR]

T-2 toxin and deoxynivalenol (DON) are type A and B trichothecenes, respectively. They widely occur as pollutants in food and crops and cause a series of toxicities, including immunotoxicity, hepatotoxicity, and neurotoxicity. Oxidative stress is the primary mechanistic basis of these toxic effects. Increasing amounts of evidence have shown that mitochondria are significant targets of apoptosis caused by T-2 toxin- and DON-induced oxidative stress via regulation of Bax/B-cell lymphoma-2 and caspase-3/caspase-9 signaling. DNA methylation and autophagy are involved in oxidative stress related to apoptosis, and hypoxia and immune evasion are related to oxidative stress in this context. Hypoxia induces oxidative stress by stimulating mitochondrial reactive oxygen species production and regulates the expression of cytokines, such as interleukin-1β and tumor necrosis factor-α. Programmed cell death-ligand 1 is upregulated by these cytokines and by hypoxia-inducible factor-1, which allows it to bind to programmed cell death-1 to enable escape of immune cell surveillance and achievement of immune evasion. This review concentrates on novel findings regarding the oxidative stress mechanisms of the trichothecenes T-2 toxin and DON. Importantly, we discuss the new evidence regarding the connection of hypoxia and immune evasion with oxidative stress in this context. Finally, the trinity of hypoxia, oxidative stress and immune evasion is highlighted. This work will be conducive to an improved understanding of the oxidative stress caused by trichothecene mycotoxins. [ABSTRACT FROM AUTHOR]

Ladle slag entrainment during the last stages of ladle draining contaminates the liquid steel in the tundish. Another effect of ladle slag is that it contaminates the tundish covering and hinders it to perform its intended functions. Affected area of tundish covering by ladle slag entrainment was quantified using a first principle-based mechanistic model in this study. Ladle slag hits the bottom of the tundish and bounces back towards the tundish cover and forms a plume-like structure. A reduced scale model was developed, and the phenomena of ladle slag entrainment were monitored using a high-speed high-resolution camera along with shadowgraphy. Slag plume velocity was measured automatically using an image processing software, ImageJ. Plume velocity was found to be reducing during last stages of entrainment. Total affected area consists of a primary area and a secondary extension area. Diameters of the primary, secondary, and total affected area are calculated using a first principle-based mechanistic model and validated with the physical modeling experiments. Additionally, the pushing of tundish slag by the ladle slag was observed in one of the four distinct stages of ladle draining condition. [ABSTRACT FROM AUTHOR]

Mycotoxins and pesticides regularly co-occur in agricultural products worldwide. Thus, humans can be exposed to both toxic contaminants and pesticides simultaneously, and multi-methods assessing the occurrence of various food contaminants and residues in a single method are necessary. A two-dimensional high performance liquid chromatography tandem mass spectrometry method for the analysis of 40 (modified) mycotoxins, two plant growth regulators, two tropane alkaloids, and 334 pesticides in cereals was developed. After an acetonitrile/water/formic acid (79:20:1, v/v/v) multi-analyte extraction procedure, extracts were injected into the two-dimensional setup, and an online clean-up was performed. The method was validated according to Commission Decision (EC) no. 657/2002 and document N° SANTE/12682/2019. Good linearity (R2 > 0.96), recovery data between 70-120%, repeatability and reproducibility values < 20%, and expanded measurement uncertainties < 50% were obtained for a wide range of analytes, including very polar substances like deoxynivalenol-3-glucoside and methamidophos. However, results for fumonisins, zearalenone-14,16-disulfate, acid-labile pesticides, and carbamates were unsatisfying. Limits of quantification meeting maximum (residue) limits were achieved for most analytes. Matrix effects varied highly (−85 to +1574%) and were mainly observed for analytes eluting in the first dimension and early-eluting analytes in the second dimension. The application of the method demonstrated the co-occurrence of different types of cereals with 28 toxins and pesticides. Overall, 86% of the samples showed positive findings with at least one mycotoxin, plant growth regulator, or pesticide. [ABSTRACT FROM AUTHOR]

The most common mycotoxin found in European foodstuffs, especially unprocessed grains, is deoxynivalenol (DON), which inhibits proteosynthesis and induces oxidative stress. The sorption properties of humic acids (HA) indicate a promising potential in the reduction of mycotoxin intoxication. In an experimental biomodel lasting 35 days, we investigated the effect of a dietary supplement containing HA on the antioxidant status in the liver, heart and kidney mitochondria and in the blood plasma of rats exposed to the stress factor of orally administered DON. DON was administered at doses of 100 and 200% above the maximum tolerable daily intake (1.0 μg/kg body weight/day). We evaluated the activities of the following enzymes: superoxide dismutase, glutathione peroxidase, glutathione reductase, glutathione-S-transferase and tripeptide glutathione. Administration of DON has been shown to induce oxidative stress, the intensity of which appears to be directly dependent on the concentration of mycotoxin. Supplementation with 1% HA (10 g/kg of feed) resulted in alleviation of this stress, in which the preventive effect of HA may have been implicated partly by affecting the adsorption of DON through the GIT and limiting its bioavailability. There were also signs that it can act by actively interfering with ROS inhibition to help achieve redox homeostasis. However, there is another aspect that deserves attention, namely, the metabolism of HA. The results presented in our work suggest that the mechanism of possible metabolism occurs predominantly in the liver, including an oxidative increase in polarity and subsequent conjugation reactions mediated by the GST-GSH system. [ABSTRACT FROM AUTHOR]

Lateral flow immunoassays (LFIAs) are widely used for rapid food safety screening analysis. Thanks to simplified protocols and smartphone readouts, LFIAs are expected to be increasingly used on-site, even by non-experts. As a typical follow-up in EU regulatory settings, suspect samples are sent to laboratories for confirmatory analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS). However, re-analysis by LC-MS/MS is laborious and time-consuming. In this work, an identification LFIA (ID-LFIA) approach followed by quadrupole-orbitrap MS or triple quadrupole MS/MS analysis is presented. As a proof of concept, a dedicated ID-LFIA strip was developed for the mycotoxin deoxynivalenol (DON) following its initial screening by a commercial smartphone LFIA. The ID-LFIA strip can be simply immersed in the same sample extract used for the smartphone LFIA screening, and next, DON is retrieved from the monoclonal antibody with a dissociation solution consisting of methanol/ammonia. The solution thus obtained was analyzed directly in MS in order to rapidly confirm the presence of DON and any cross-reacting species. The protocol developed is capable of coping with severe ion suppression caused by LFIA buffers and nitrocellulose substrate residues. Initial analysis of blank, spiked, and incurred samples showed that the newly developed ID-LFIA-MS method was able to confirm the presence or absence of mycotoxins in the samples previously analyzed by LFIA and also differentiate between DON and DON 3-glucoside yielding the positive screening result. The concept and technique developed are envisaged to complement on-site screening and confirmation of any low molecular weight contaminant in future food control frameworks. [ABSTRACT FROM AUTHOR]

This study aimed to establish a purifying immunoaffinity column (IAC) for four kinds of Fusarium toxins (zearalenone, deoxynivalenol, T-2, and HT-2) simultaneously using poly-glycidyl methacrylate–divinylbenzene [P(GMA–DVB)] as the column matrix. The P(GMA–DVB) microsphere was synthesized using glycidyl methacrylate (GMA) and diethylbenzene (DVB) at an optimal polymerization temperature of 80 °C, the initiator 2,2′-azodiisobutyronitrile concentration of 2.50 g/mL H2O, and the DVB-to-GMA ratio of 3:5 (wt.). The polymerized P(GMA–DVB) microspheres were modified with 1,6-hexanediamine, activated with glutaraldehyde, and coupled with the four toxin monoclonal antibodies. The coupling rate of the activated resin was calculated to be 95.95%. The maximum capacity of the IACs was 264 ng deoxynivalenol, 231 ng T-2, 210 ng HT-2, and 292 ng zearalenone. After eight uses, the maximum capacity had no obvious decrease. The recovery of the four Fusarium toxins on the P(GMA–DVB) IAC was 70.03–85.39% in flour, with the relative standard deviations of 4.67–15.93%. The IACs were applied to the actual flour samples. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is commonly found in wheat and wheat-derived foods, posing a threat to human health. Biodegradation is an efficient and eco-friendly measure for mycotoxin detoxification. Understanding the mechanism of DON biodegradation is hence of great importance. Herein, we report the application of metabolomics methods for the analysis of DON degradation by a bacterial consortium isolated from wheat leaves collected in Jiangsu Province. Metabolomics analysis combined with a nuclear magnetic resonance analysis revealed the main degradation product, 3-keto-DON, and a minor degradation product, 3-epi-DON. Further study illustrated that DON underwent a two-step epimerization through the intermediate 3-keto-DON. Sequencing analysis of the 16S rRNA metagenome of the microorganismal community suggested that the abundance of three bacterial genera, Achromobacter, Sphingopyxis, and Sphingomonas, substantially increased during the coculture of bacterial consortium and DON. Further investigation revealed that Devosia sp. might be responsible for the epimerization of 3-keto-DON. These findings shed light on the catabolic pathways of DON during biodegradation and illustrate the potential of using metabolomics approaches in biodegradation studies. Key Points • A bacterial consortium was isolated with good deoxynivalenol-degrading potential. • Metabolomics approaches were successfully used to interpret the degradation pathway. • A trace-amount degradation product was determined by metabolomics and NMR analysis. [ABSTRACT FROM AUTHOR]

Enoyl-CoA hydratase (Ech) is an important and well-recognized enzyme that functions in the degradation of fatty acids by β-oxidation. However, its functions in plant pathogenic fungi are not well known. We characterized an Ech1 orthologue, FgEch1, in Fusarium graminearum. The FgEch1 deletion mutant was defective in the utilization of short-chain fatty acids and conidiation, but not in hyphal growth on glucose-rich media or in perithecium formation. The FgEch1 deletion mutant showed reduced deoxynivalenol (DON) production and virulence in plants. Deletion of FgEch1 also led to increased production of lipid droplets and autophagy. FgEch1, which was localized in the mitochondrion, required the MTS domain for mitochondrial localization and function in F. graminearum. Taken together, these data indicate that mitochondrial FgEch1 is important for conidiation, DON production, and plant infection. [ABSTRACT FROM AUTHOR]

An aptamer-based assay for the determination of two different kinds of fusarium mycotoxins, i.e., zearalenone (ZEN) and fumonisin B1 (FB1), is presented. Based on the inner filter effect (IFE) strategy, the contents of ZEN and FB1 can be simultaneously quantified. It is making use of 65-nm gold nanorods (AuNRs), 20-nm upconversion nanoparticles (UCNPs), fluorescence dyes, and DNA sequences. In the absence of ZEN and FB1, the UCNPs and AuNRs associate through DNA sequences. Due to IFE effect, weak fluorescence signals are collected. In the presence of ZEN or FB1, UCNPs and AuNRs become unstable and partially separate from each other. This results in the recovery of fluorescence signals. Under 980-nm laser excitation, the logarithmic values of fluorescence signal intensities at 606 nm and 753 nm gradually increase with the concentration of ZEN and FB1 in the ranges 0.05–100 μg L−1 (the coefficient of determination is 0.997) and 0.01–100 ng L−1 (the coefficient of determination is 0.986), respectively. The limits of detection (LOD) of the fabricated assay for ZEN and FB1 are 0.01 μg L−1 and 0.003 ng L−1, respectively. The proposed method has a high selectivity over other competitive mycotoxins, including aflatoxin B1, ochratoxin A, patulin and ochratoxin B. The applicability of the assay was evaluated in the determination of ZEN and FB1 contents in spiked corn samples. The average recoveries ranged from 89.9 to 106.6%. This result confirms the practicality of this method. [ABSTRACT FROM AUTHOR]

Over the past several years, numerous studies have demonstrated the feasibility of using engineered nanoparticles as antifungals, especially against those fungal pathogens that produce mycotoxins and infect plants, animals, and humans. The high dosage of nanoparticles has been a concern in such antifungal applications due to the potential toxicological and ecotoxicological impacts. To address such concerns, we have recently introduced the idea of inhibiting mycotoxin biosynthesis using low doses of engineered nanoparticles. At such low doses these particles are minimally toxic to humans and the environment. From our studies we realize that for the effective use of nanotechnology to intervene in the biology of fungal pathogens and for an accurate evaluation of the impacts of the increasingly growing nanomaterials in the environment on fungi and their interacting biotic partners, there is a pressing need for a rigorous understanding of nano-fungal interactions, which is currently far from complete. In this minireview, we build on the available evidence from nano-bio interaction research and our recent interaction studies with Aspergillus cells and engineered silver nanoparticles to introduce a potential theoretical model for nano-fungal interactions. The aim of the proposed model is to provide an initial insight on how nanoparticle uptake and their transformation inside fungal cells, possibly influence the production of mycotoxins and other secondary metabolites of filamentous fungi. [ABSTRACT FROM AUTHOR]

Avena spp. were artificially inoculated with Fusarium culmorum Sacc. (FC) and F. graminearum Schwabe (FG) causing Fusarium head blight (FHB). This disease is often accompanied by the accumulation of Fusarium mycotoxins mainly deoxynivalenol (DON) in grains. The contaminated grains with this mycotoxin are toxic to their consumers. Genotypes Avena spp. with low DON accumulation in grains can be used as genetic resources suitable for the formation of new resistant varieties of oats against FHB caused by fungi FC and FG. The aim was to find out and to compare the potential for mycotoxin accumulation in grains between genotypes of Avena spp. after artificial inoculation panicles by FC and FG using three spray methods and identify genotypes of Avena (A. abyssinica, A. byzantina, A. canariensis, A. fatua, A. ludoviciana, A. nuda, A. sativa, A. sterilis, A. strigosa) with low toxin accumulation in whole grain. The average accumulations of DON in the grains of Avena spp. gradually increased from the spray inoculation (0.68 mg · kg−1), spray + polyethylene (PE) bag cover 24 hrs (2.75 mg · kg−1) and spray + PE bag/48 hrs. (9.46 mg · kg−1) methods. We found out that after application of each used method, the high DON accumulation in grains was found in A. canariensis, and low DON levels were found in A. byzantine and A. sterilis. [ABSTRACT FROM AUTHOR]

Allogenic organic matter (AOM) composed of extracellular and intracellular organic matter (EOM and IOM) is a major precursor of halogenated carbonaceous and nitrogenous disinfection by-products (C-DBPs and N-DBPs) upon chlorination. The EOM and IOM extracted from Microcystis aeruginosa were analyzed based on bulk parameters and organic fractions with different molecular weight by liquid chromatography with organic carbon detection (LC-OCD). It investigated the efficiency of a conventional gravity system (CGS) and dissolved air flotation (DAF) in the removal of organic precursors, together with measurement of the formation of four major trihalomethanes (THMs) and haloacetonitriles (HANs) in treated water upon chlorination. The results showed that EOM accounted for 59% of building blocks and humic substances, whereas for IOM, 54% were low molecular weight (LMW) neutrals. Both CGS and DAF showed 57–59% removal of dissolved organic carbon (DOC) from EOM and IOM. Regarding DON removal, DAF was found to be more effective, i.e., 8% higher than CGS for EOM. Moreover, the removal of LMW acids and neutrals (not easy to remove and are major precursors of DBPs) from EOM and IOM by DAF was higher than from CGS. The amounts of DBPs measured in all the samples treated for interchlorination were much lower than in the samples for prechlorination. Although the precursors of EOM had a higher concentration than in IOM, THMs and HANs were detected for IOM at a higher concentration, which might be attributed to higher amounts of aromatic, aliphatic moisture and protein compounds in the IOM. Comparatively, DAF showed lower THM and HAN values than CGS water, particularly for IOM. Also, DAF showed a sharp decrease in THMs and an insignificant increase in HANs according to time. [ABSTRACT FROM AUTHOR]

Because I Fusarium i mycotoxins are produced on crops in the field, it is crucial to understand how agricultural practices affect mycotoxin contamination in grain (Nakajima [7]). In this review, I present the current understanding of GAP to control mycotoxin contamination with FHB in wheat and barley. This timing is reasonable for wheat because wheat is most susceptible to FHB during anthesis, when anthers are extruded from the florets and are commonly the initial site of I Fusarium i infection in wheat. These results suggest that the optimal time for chemical control of FHB and mycotoxin contamination in cleistogamous barley is around the beginning of SAE, rather than at anthesis (Nakajima et al. [8]; Yoshida et al. [17]). [Extracted from the article]

Zearalenone (ZEA), a mycotoxin produced by several Fusarium spp., is most commonly found as a contaminant in stored grain. ZEA derivatives (α-zearalenol (α-ZOL), β-zearalenol (β-ZOL)) can also be produced by Fusarium spp. in corn stems infected by fungi in the field. Also, following oral exposure, zearalenone is metabolized in various tissues, particularly in the liver, the major metabolites being α-ZOL and β-ZOL. The co-exposure of cells to mixture of a combination of mycotoxins may cause an increase of toxicity produced by these mycotoxins. In this in vitro study, we investigated the combined effects of ZEA, α-ZOL, β-ZOL in binary mixtures on the viability and inflammatory response of human liver cancer cell line (HepG2). Cell viability was assessed after 72 h using a neutral red assay. Effect of the toxins and their binary combinations on the expression of genes involved in inflammation (IL-1β, TNF-α, and IL-8) were assessed through qPCR. Our viability data showed that irrespective of the toxin combinations, the toxins have synergistic effect. ZEA + α-ZOL and ZEA + β-ZOL mixtures have induced a slight to high antagonistic response on inflammatory cytokines at low concentrations that have turned into strong synergism for high concentrations. α-ZOL + β-ZOL showed antagonistic effects on inflammation for IL-1β and TNF-α, but act synergic for IL-8 at high toxin concentrations. This study clearly shows that co-contamination of food and feed with ZEA metabolites should be taken into consideration, as the co-exposure to mycotoxins might result in stronger adverse effect than resulted from the exposure to individual toxin. [ABSTRACT FROM AUTHOR]

Peroxisomes are indispensable organelles that play critical roles in various biological processes in eukaryotic cells. PEX4, one of the peroxins, is the ubiquitin-conjugating enzyme. To functionally characterize roles of FgPEX4 in the phytopathogenic fungus, Fusarium graminearum, we constructed a deletion mutant of FgPEX4 (ΔPEX4) through homologous recombination. ΔPEX4 displayed reduced mycelial growth, conidiation, and the production of perithecia. ΔPEX4 was defective in pathogenicity and production of the mycotoxin deoxynivalenol (DON). In addition, FgPEX4 was involved in cell wall integrity, lipid droplet accumulation, and the elimination of reactive oxygen species. Western blot analysis revealed reduced phosphorylation of Mgv1 in the ∆PEX4 mutant. Importantly, proteomics analysis indicated that protein expression levels related to protein biosynthesis, fatty acid metabolism, cell wall synthesis, and oxidation–reduction reactions were downregulated in ΔPEX4 compared with the wild type. Taken together, these results demonstrate that FgPEX4 is important for development, pathogenicity, and cell wall integrity. [ABSTRACT FROM AUTHOR]

Ethanol (EtOH) is one of the most frequently abused drugs with heavy health, economic, and societal burdens. Although moderate to low EtOH may have some neuroprotective effects, heavy EtOH consumption associated with high blood alcohol level (BAL) can be quite detrimental. The brain is particularly vulnerable to the damaging effects of high BAL, leading to neuronal loss, cognitive, and behavioral deficits. Although the exact causes of these detriments are not fully elucidated, it is believed that damage to the cholinergic system is at least partially responsible for the cognitive impairment. Thus, high BAL may result in selective apoptotic damage to the cholinergic neurons. Donepezil (DON), a centrally acting, reversible and non-competitive acetylcholinesterase (AChE) inhibitor, approved for use in Alzheimer’s disease (AD), may also attenuate EtOH-induced cognitive impairment. Cognitive effects of DON might be due to an anti-apoptotic activity as some AChE inhibitors have been shown to have this property. The aim of this study was to determine whether DON might protect against EtOH-induced toxicity and whether such protection might be apoptotically mediated. We exposed the human neuroblastoma-derived, SH-SY5Y cells to a relatively high concentration of EtOH (500 mM) for 24 h and evaluated the effects of two concentrations of DON (0.1 and 1.0 μM) on alcohol-induced toxicity and caspase-3, an apoptotic marker. We found a dose-dependent protection of DON against EtOH-induced toxicity as well as dose-dependent attenuation of EtOH-induced increases in caspase-3 levels. Thus, DON may inhibit apoptosis as well as alcohol-induced toxicity. [ABSTRACT FROM AUTHOR]

Fusarium graminearum and F. culmorum are phytopathogenic species that cause head blight and crown rot diseases in cereals worldwide. These two phytopathogenic fungi show great variation in terms of phenotypic, biochemical and genetic characteristics. In this study, linear growth rate, thiophanate methyl (TPM) resistance, survival and toxin production- associated gene expressions, and beta-tubulin nucleotide sequence knowledge of F. graminearum from Turkey and Iran and F. culmorum isolates from Turkey were investigated. The mean linear growth rate values of isolates ranged from 7.58 to 15.16 mm/day. In TPM resistance analysis, each isolate was characterized as highly sensitive to TPM, with IC50 values between 2 and 4 μg/ml TPM. TPM-subjected isolates showed decreased tri5 gene expression and increased Mgv and StuA expression. Amino acid conservations at codons 198 and 200 in the beta tubulin gene were also investigated in order to confirm TPM resistance and reveal genetic diversity in Fusarium species. Each isolate showed conserved amino acids at 198 and 200 codons, resulting in sensitive genotypes. The bootstrap support value for genetic similarity was more than 70%. Findings obtained from this study showed that Fusarium isolates that cause head blight and crown rot in Turkey have a low level of variation in terms of phenotype and genetic characteristics. Moreover, this is the first report to show that TPM led to a potential decrease in toxin production-associated genes and increased potential of sexual/asexual reproduction genes. Results could provide improved and comprehensive data for the struggle against Fusarium pathogens. [ABSTRACT FROM AUTHOR]