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Background: Food safety has become a serious global concern. Therefore, there is a need for effective detection technologies in this field. Currently, the development of effective on-site detection techniques is extremely important for food safety. However, the traditional on-site detection methods currently lack effective signal amplification. Herein, the aim of this study was to construct a nanopore-based label-free CRISPR/Cas12a system for the detection of Zearalenone (ZEN). The method is expected to be highly sensitive for portable detection of ZEN in food., Results: The proposed strategy was mainly involved three steps, including the displacement of the target DNA, the triggering of the cleavage of hairpin DNA probes (probes 1) by the trans-cleavage of CRISPR/Cas12a, and the generation of a measurable nanopore current signal. The probes 1 and DNA after the cleavage of probes 1 (probes 2) produce different characteristic nanopore signals as they pass through the nanopore. The established method achieved a low limit of detection (LOD) of 6.52 fM for ZEN and a wide liner range under optimized conditions. Furthermore, the practical applicability of this method was verified in real maize samples and showed good recoveries (90.68-101.98 %) and low relative standard deviations (RSD) (9.21-9.72 %). Therefore, this method is a promising option for rapid and ultrasensitive detection of ZEN., Significance and Novelty: The study presented a portable nanopore-based CRISPR/Cas12a signal amplification detection system for the detection of ZEN in food, which had a low LOD and the advantages of rapid, portability, and on-site detection potential. In conclusion, the method presented a promising prospect and universal platform for the detection of ZEN and other mycotoxins, offering a novel insight into on-site food safety detection., Competing Interests: Declaration of competing interest The authors declare that they do not have any competing financial or personal interests which can influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Zearalenone (ZEN) is a mycotoxin that poses significant risks to human and animal health due to its mutagenic, immunosuppressive, and carcinogenic properties. This study presents a novel analytical method for detecting ZEN using electrochemical impedance spectroscopy (EIS) combined with a molecularly imprinted polymer (MIP). ZEN, used as the template molecule, was incorporated into polypyrrole on screen-printed electrodes (SPE), and a ZEN-sensitive MIP sensor was created through template removal. The modified sensor surfaces were characterized by EIS and scanning electron microscopy (SEM). An impedimetric MIP sensor for ZEN was developed, offering a detection range from 1 pM to 500 pM. The method's limit of detection (LOD) was established at 1 pM (0.3 pg/mL) with a signal-to-noise ratio of 3 (S/N = 3). The method demonstrated high precision and accuracy, with a maximum relative standard deviation (RSD) of less than 4.4% at a 95% confidence level, and relative error (RE) values ranging from -0.8% to -2.7%. The selectivity of the developed MIP sensor was evaluated using ochratoxin A, ochratoxin B, and aflatoxin B1, with no significant interference observed. ZEN recovery from spiked samples was between 95% and 105%, indicating that the method was successfully applied to grain samples, including corn, rice, and wheat., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Rice plays an important role in the daily diet in China and therefore its quality and safety have been of great concern. However, few systematic studies have investigated Fusarium community and toxins in rice grains. Here, we collected 1381 rice samples from Jiangsu Province in eastern China and found a higher frequency of zearalenone (ZEN), deoxynivalenol (DON), fumonisins (FBs), and beauvericin (BEA). The positive samples were individually contaminated with a minimum of one and a maximum of ten toxins. Fusarium was isolated and identified as the major fungus, which exhibited temporal and geographical distribution. The most prevalent species complexes within this genus were Fusarium incarnatum-equiseti species complex (FIESC), Fusarium fujikuroi species complex (FFSC), and Fusarium sambucinum species complex (FSAMSC). Nevertheless, the amplicon sequence analysis revealed a low relative abundance of Fusarium in the rice panicles, and the fungal community exhibited an irregular change along with the symptom's emergence. In vitro toxigenic profiles of Fusarium strains showed significant complexity and specificity depending on the type and content. FIESC strains were non-pathogenic to wheat heads and weakly pathogenic to maize ears, respectively, accumulating lower amounts of toxins than F. asiaticum and F. fujikuroi. There was no significant variation in the ability to cause panicle blight in rice among the various species tested. Our study provides detailed information about the contamination of Fusarium toxins and community in rice after harvest. This information is valuable for understanding the relationship between Fusarium and rice and for developing effective control strategies., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jianhong Xu reports financial support was provided by Jiangsu Agriculture Science and Technology. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The fungal genus Fusarium contains many toxigenic pathogens of maize with associated yield losses, reduction of grain quality, and accumulation of mycotoxins in harvested grains. To determine zearalenone (ZEN) concentration and identify the various Fusarium species in commercial maize grains, a survey of 75 maize samples, collected from 11 market centers in the five regions in northern Ghana was identified based on morphological characteristics, sequence analysis of the internal transcribed spacer region, and polymerase chain reaction using species-specific primers. ZEN levels were determined using HPLC. ZEN contamination was recorded in 33.3% of the maize samples, with concentrations ranging from 0.61 to 3.05 µg/kg. Based on VERT1/2 and TEF 1-α sequencing, F. verticillioides was the most prevalent species in the studied samples: 40.35% from the Upper East Region, 28.07% from the North East Region, 19.30% from the Upper West Region, 10.53% from the Savannah Region, and 1.75% for the Northern Region. Other fungal species found were F. equiseti and F. solani. A higher number of the Fusarium isolates were found in white maize (609 isolates from 27 samples) compared to yellow maize (225 isolates from 23 samples)., (© 2024. The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.)

The debranning process, at an industrial scale, was applied to grains of two wheat cultivars to determine its effect on Fusarium mycotoxin content and antioxidant activity. Grain samples from the BRS Marcante and BRS Reponte wheat cultivars, naturally contaminated by Fusarium, were used in the study. The dry wheat samples were processed on the polisher once or twice and evaluated by hardness index, chemical composition (moisture, protein, and ash), deoxynivalenol (DON) and zearalenone (ZON) levels, phenolic content, and antioxidant activity. In the BRS Marcante cultivar, the debranning process only slightly reduced the DON and ZON contents in whole-wheat flours compared with the previous cleaning treatment (no-debranned). In the BRS Reponte cultivar, the DON concentration decreased by 36% at a debranning ratio of 5%, obtained by polishing, compared with prior cleaning treatment (no-debranned). In addition, the polishing reduced the ZON level by 56% compared with the cleaned wheat. The debranning process did not reduce the antioxidant capacity. Therefore, debranning is a suitable technology to obtain safer and healthier food by minimizing the mycotoxin content and retaining antioxidant capacity., (© 2024. The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.)

A novel approach for the simultaneous separation of zearalenone (ZEN) and four types of aflatoxins (AFB1, AFB2, AFG1 and AFG2) from rice samples was presented. This approach utilized modified MIL-101(Cr)-NH2 as core, with molecularly imprinted polymers (MIPs) serving as the shell. The MIL-101(Cr)-NH2 was prepared via ring-opening reaction, while the imprinted polymers were synthesized using warfarin and 4-methylumbelliferyl acetate as co-pseudo template, ethylene glycol dimethacrylate as the cross-linker and azobisisobutyronitrile as initiator. The resulting co-pseudo-template-MIPs (CPT-MIPs) were thoroughly characterized and evaluated. Adsorption studies demonstrate that the adsorption process of CPT-MIPs follows a chemical monolayer adsorption mechanism, with imprinted factors ranging from 1.24 to 1.52 and selective factors ranging from 1.29 to 1.52. Self-made columns were prepared, and the method for separation was developed and validated. The limit of detections ranged from 0.12 to 2.09 μg/kg, and the limit of qualifications ranged from 1.2 to 6.25 μg/kg. To assess the reliability of the method, ZEN and AFs were spiked at three different levels, and the recoveries ranged from 79.53 to 94.58%, with relative standard deviations of 2.90-5.78%., (© The Author(s) 2024. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Environmental factors influence fungal growth and mycotoxin production in stored grains. However, the concentrations of free mycotoxins and their conjugates and how they are impacted by different interacting environment conditions have not been previously examined. The objectives of this study were to examine the impact of storage conditions (0.93-0.98 a w ) and temperature (20-25 °C) on (a) the concentrations of deoxynivalenol and zearalenone and their respective glucosides/conjugates and (b) the concentrations of emerging mycotoxins in both naturally contaminated and irradiated wheat grains inoculated with Fusarium graminearum. Contaminated samples were analysed for multiple mycotoxins using Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). Method validation was performed according to the acceptable performance criteria set and updated by the European Commission regulations No. 2021/808/EC. As an important conjugate of deoxynivalenol, the concentrations of deoxynivalenol-3-glucoside were significantly different from its precursor deoxynivalenol at 0.93 a w (22% moisture content- MC) at 25 °C in the naturally contaminated wheat with a ratio proportion of 56:44% respectively. The high concentrations of deoxynivalenol-3-glucoside could be influenced by the wheat's variety and/or harvested season/fungal strain type/location. Zeralenone-14-sulfate concentrations were surprisingly three times higher than Zearalenone in the naturally contaminated wheat at 0.98 a w (26% MC) at both temperatures. Emerging mycotoxins such as moniliformin increased with temperature rise with the highest concentrations at 0.95 a w and 25 °C. These findings highlight the influence and importance of storage a w x temperature conditions on the relative presence of free vs conjugated mycotoxins which can have implications for food safety., (© 2024. The Author(s).)

Driven by their exceptional optical characteristics, robust chemical stability, and facile bioconjugation, gold nanoparticles (AuNPs) have emerged as a preferred material for detection and biosensing applications in scientific research. This study involves the development of a simple, rapid, and cost-effective colorimetric immuno-sensing probe to detect aflatoxin B1 and zearalenone using AuNP antibody (AuNP-mAb) conjugates. Anti-toxin antibodies were attached to the AuNPs by using the physical adsorption method. The colorimetric immunosensor developed operates on the principle that the optical properties of the AuNP are very sensitive to aggregation, which can be induced by a critical high salt concentration. Although the presence of antibodies on the AuNP surface inhibits the aggregation, these antibodies bind to the toxin with higher affinity, which leads to exposure of the surface of AuNPs and aggregation in a salt environment. The aggregation triggers a noticeable but variable alteration in color from red to purple and blueish gray, as a result of a red shift in the surface plasmon resonance band of the AuNPs. The extent of the shift is dependent on the toxin exposure dose and can be quantified using a calibration curve through UV-Visible-NIR spectroscopy. The limit of detection using this assay was determined to be as low as 0.15 ng/mL for both zearalenone and aflatoxin B1. The specificity of the prepared immunoprobe was analyzed for a particular mycotoxin in the presence of other mycotoxins. The developed immunoprobe was evaluated for real-world applicability using artificially spiked samples. This colorimetric immunoprobe based on localized surface plasmon resonance (LSPR) has a reduced detection limit compared to other immunoassays, a rapid readout, low cost, and facile fabrication.

Dielectric barrier discharge ionization is increasingly used for rapid detection in ambient mass spectrometry, although more often for gaseous and highly volatile samples than for solids and liquids. In this project, we present a rapid and sensitive method for detecting mycotoxins and demonstrate its capability for the detection of aflatoxin B1, zearalenone, and ochratoxin A in food samples. Our method is based on thermal desorption coupled to dielectric barrier discharge ionization mass spectrometry (TD-DBDI-MS), which we show generates minimal interferences and produces almost exclusively molecular ions. We detected mycotoxins in various food samples, including corn, peanuts, millet, and rice. Our method has a linear dynamic range of 1 μg kg -1 to 100 μg kg -1 for all three mycotoxins and a limit of detection (LOD) of 0.31 μg kg -1 , 0.28 μg kg -1 and 0.43 μg kg -1 , respectively. It is simple, rapid, reduces the pretreatment steps and has significant potential for practical applications.

Zearalenone, a prominent mycotoxin produced by Fusarium spp., ubiquitously contaminates cereal grains and animal feedstuffs. The thermal stability of zearalenone creates serious obstacles for traditional removal methods, which may introduce new safety issues, or reducing nutritional quality. In contrast, biological technologies provide appealing benefits such as easy to apply and effective, with low toxicity byproducts. Thus, this review aims to describe the occurrence of zearalenone in cereals and cereal-based feedstuffs in the recent 5 years, outline the rules and regulations regarding zearalenone in the major countries, and discuss the recent developments of biological methods for controlling zearalenone in cereals and cereal-based feedstuffs. In addition, this article also reviews the application and the development trend of biological strategies for removal zearalenone in cereals and cereal-based feedstuffs.

We conducted a comprehensive examination of liquid mycotoxin reference standards. A total of 30 different standards were tested, each containing 10 samples of three distinct substances: Aflatoxin B1, Deoxynivalenol, and Zearalenone. The standards were sourced from 10 different global market leading manufacturers. To facilitate comparison, all the standard sets were adjusted to the same concentration level. The standards were analyzed using the techniques LC-MS/MS, HPLC-DAD, and LC-HRMS to assess their quality attributes. Regarding the validation of the reference values, it was observed that 30% of the suppliers provided reference standards that were either below the lower acceptance limit or above the higher acceptance limit, confirmed by both the LC-MS/MS and HPLC-DAD methods. Furthermore, a total of 12 impurities were found in the DON standards, 10 in the AFB 1 standards, and 8 in the ZON standards, distributed across all the suppliers. Therefore, this study suggests relevant adjustments to the ISO 17034 standard, proposing that the purity of a raw material should be uniformly based on q-NMR analysis, as most manufacturers state the purity of their certificates is determined using HPLC-UV or LC-MS/MS. Liquid standards with a shelf life of ≤1 year should not exceed an uncertainty of 3%. Standards that have a longer shelf life should not have more than 5% uncertainty. This study also emphasizes the importance of stability. The standards should undergo continuous long-term monitoring; otherwise, products may exhibit a target value of only 80%, as seen in one instance. It is also recommended to include proof of HPLC and LC-MS/MS analyses on the certificate of each released batch of a final product.

In this work, a potential-controlled electrochromic visual biosensor was developed for detecting zearalenone (ZEN) using a distance readout strategy. The sensor chip includes a square detection area and a folded signal output area created with laser etching technology. The detection area is modified with graphene oxide and ZEN aptamer, while Prussian blue (PB) is electrodeposited onto the signal output channel. When an appropriate voltage is applied, PB in the signal output area is reduced to colorless Prussian white (PW). The target ZEN molecules have the capability to release aptamers from graphene oxide (GO) surface in the detection area, resulting in a subsequent change in the potential of the visual signal output channel. This change determines the length of the channel that changes from blue to colorless, with the color change distance being proportional to the ZEN concentration. Using this distance readout strategy, ZEN detection within the range of 1 ng/mL to 300 ng/mL was achieved, with a detection limit of 0.29 ng/mL., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Maize grain samples collected from 129 small-scale farmers' stores in southern and southwestern Ethiopia were analysed by LC-MS/MS for a total of 218 mycotoxins and other fungal metabolites of which 15% were regulated mycotoxins. Mycotoxins produced by Penicillium , Aspergillus , and Fusarium accounted for 31%, 17%, and 12% of the metabolites, respectively. Most of the current samples were contaminated by masked and/or emerging mycotoxins with moniliformin being the most prevalent one, contaminating 93% of the samples. Each sample was co-contaminated by 3 to 114 mycotoxins/fungal metabolites. Zearalenone, fumonisin B1, and deoxynivalenol were the dominant mycotoxins, occurring in 78%, 61%, and 55% of the samples with mean concentrations of 243, 429, and 530 µg/kg, respectively. The widespread co-occurrence of several mycotoxins in the samples may pose serious health risks due to synergistic/additional effects.

The contamination of mycotoxins poses a serious threat to global food security, hence the urgent need for simultaneous detection of multiple mycotoxins. Herein, two SERS nanoprobes were synthesized by embedded SERS tags (4-mercaptopyridine, 4MPy; 4-mercaptobenzonitrile, TBN) into the Au and Ag core-shell structure, and each was coupled with the aptamers specific to ochratoxin A (OTA) and zearalenone (ZEN). Meanwhile, a rigid enhanced substrate Indium tin oxide glass/AuNPs/Graphene oxide (ITO/AuNPs/GO) was combined with aptamer functionalized Au@AgNPs via π-π stacking interactions between the aptamer and GO to construct a surface-enhanced Raman spectroscopy (SERS) aptasensor, thereby inducing a SERS enhancement effect for the effective and swift simultaneous detection of both OTA and ZEN. The presence of OTA and ZEN caused signal probes dissociation, resulting in an inverse correlation between Raman signal intensity (1005 cm -1 and 2227 cm -1 ) and the concentrations of OTA and ZEN, respectively. The SERS aptasensor exhibited wide linear detection ranges of 0.001-20 ng/mL for OTA and 0.1-100 ng/mL for ZEN, with low detection limits (LOD) of 0.94 pg/mL for OTA and 59 pg/mL for ZEN. Furthermore, the developed SERS aptasensor demonstrated feasible applicability in the detection of OTA and ZEN in maize, showcasing its substantial potential for practical implementation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A refined electrochemical aptamer sensing technique using PEI@Ti-MOF@Ti 3 C 2 Tx-MXene was developed for the sensitive detection of ZEN in food samples. A titanium-based metal-organic skeleton (NH 2 -MIL-125) was synthesized in situ using 2-aminoterephthalic acid as the organic ligand and tetrabutyl titanate as the metal center, followed by the simultaneous hybridization of Ti 3 C 2 Tx-MXene to synthesize a Ti-MOF@Ti 3 C 2 Tx-MXene composite material. These composites were subsequently functionalized with PEI and covalently linked to form a sensing platform on gold electrodes. Integrating a metal-organic framework (MOF) with MXene materials not only improved the electrochemical properties compared to those of individual elements but also decreased the stacking effect and increased the number of binding sites for the aptamer. The limit of detection (LOD) of this sensor was 1.64 fg mL -1 . Additionally, the sensor could efficaciously detect ZEN in cornmeal and beer samples, exhibiting outstanding stability, reproducibility, and selectivity. This highlighted its effectiveness in applications in quality supervision and food safety., Competing Interests: Declaration of competing interest There are no conflicts to declare., (Copyright © 2024. Published by Elsevier Ltd.)

In the current study, a colorimetric sensor array combined with near-infrared (NIR) spectroscopy was used to quantitatively analyze zearalenone in wheat. The portable NIR spectrometer was used to scan the porphyrin reaction points of the wheat colorimetric sensor and collect spectral data. Subsequently, based on all the NIR spectral data, the two models and three feature selection algorithms are compared, and the best performance model and the best feature variable input are selected. Concurrently, the Kernel-based Extreme Learning Machine (KELM) model optimized by the two parameter optimization algorithms was compared, and the best parameter optimization algorithm was selected. Among all evaluation models, the KELM model optimized by the Competitive Adaptive Reweighted Sampling algorithm combined with the rime optimization algorithm has the best prediction effect. The predicted R P 2 is 0.9900, and the root mean square error of prediction (RMSEP) is 18.4610 μg∙kg -1 ., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In this work, a sea urchin gold nanoparticles-zearalenone aptamer- tetramethylrhodamine sensor was constructed. Sea urchin gold nanoparticles, prepared using the seed-mediated growth method, were used as Raman substrates. Nucleic acid aptamers were mainly used as specific recognition molecules. Zearalenone detection in miscellaneous beans was accomplished using the principle of conformational change in aptamer. In addition, we evaluated the linear range, sensitivity, and selectivity of our sensor. We observed that at the displacement of 814 cm -1 , for Zearalenone concentrations of 0.01-60 ng/mL, the Raman signal intensity linearly correlated with the zearalenone concentration, with a limit of detection of 0.01 ng/mL, and recoveries of 91.7% to 108.3%. The optimum detection time was 30 min. Thus, our sensor exhibited great potential in zearalenone detection in food products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

An electrochemical sensor was developed for detecting zearalenone (ZEN) based on the mimic peptide, which was screened from the library and validated by molecular simulation and electrochemical methods. The library of the mimic peptide was constructed according to the structural analysis, molecular docking, molecular dynamics and amino acid mutation. Then, the enhanced electrical signal was attributed to gold nanoparticles (AuNPs) and reduced carboxylated graphene oxide (rGO-COOH). Under the optimal conditions, the detection limit was 0.91 pg·mL -1 (S/N = 3) with a wide linear range from 0.01 ng·mL -1 to 10 ng·mL -1 . In grain samples, a good recovery rate of 84% to 105.3% was achieved, while the rate ranged from 82% to 108.8% in the commercial ELISA kits. Additionally, the electrochemical sensor exhibited the remarkable specificity, excellent stability and better reproducibility (RSD = 1.94%). This sensor has great potential for rapidly detecting ZEN in food., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Silage has been identified as a source of different microbial toxins, that may impair farm animal health and productivity as human health can also be compromised. In this sense, the aim of this study was to determine the impact of silage additives on the concentrations of deoxynivalenol (DON) and zearalenone (ZEN) mycotoxins and, eventually, to evaluate the hygienic quality of orchardgrass (Dactylis glomerata L.) silage based on the concentration of them compared to control silage. This study evaluated the influence of biological and chemical additives used in six different varieties of orchardgrass silage on DON and ZEN mycotoxin contents for the first time. The content of both fusariotoxins (DON and ZEN) in fresh matter and grass silage were below the threshold stipulated by the European Commission. The concentration of DON ranges from ~21.86 to 37.26 ng/kg, ~10.21 to 15 ng/kg, ~20.72 to 29.14 ng/kg; and ZEN range from ~3.42 to 7.87 ng/kg, ~3.85 to 8.62 ng/kg and ~2.15 to 5.08 ng/kg, in control, biological and chemical silages, respectively. In general, the biological additive was more efficient for preventing DON contamination, whereas the chemical additive was more efficient for preventing ZEN contamination in grass silage. In summary, the results obtained in this work demonstrate that biological and chemical additives can inhibit fungal growth and mycotoxin production on Dactylis glomerata L. silage and whose use could prevent animal and human diseases., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Alba-Mejía et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The label probe plays a crucial role in enhancing the sensitivity of lateral flow immunoassays. However, conventional fluorescent microspheres (FMs) have limitations due to their short fluorescence lifetime, susceptibility to background fluorescence interference, and inability to facilitate multi-component detection. In this study, carboxylate-modified Eu(III)-chelate-doped polystyrene nanobeads were employed as label probes to construct a multiple time-resolved fluorescent microsphere-based immunochromatographic test strip (TRFM-ICTS). This novel TRFM-ICTS facilitated rapid on-site quantitative detection of three mycotoxins in grains: Aflatoxin B1 (AFB1), Zearalenone (ZEN), and Deoxynivalenol (DON). The limit of detection (LOD) for AFB1, ZEN, and DON were found to be 0.03 ng/g, 0.11 ng/g, and 0.81 ng/g, respectively. Furthermore, the TRFM-ICTS demonstrated a wide detection range for AFB1 (0.05-8.1 ng/g), ZEN (0.125-25 ng/g), and DON (1.0-234 ng/g), while maintaining excellent selectivity. Notably, the test strip exhibited remarkable stability, retaining its detection capability even after storage at 4 °C for over one year. Importantly, the detection of these mycotoxins relied solely on simple manual operations, and with a portable reader, on-site detection could be accomplished within 20 min. This TRFM-ICTS presents a promising solution for sensitive on-site mycotoxin detection, suitable for practical application in various settings due to its sensitivity, accuracy, simplicity, and portability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

To address the food safety issues caused by toxins, we established a fluorescent copper nanocluster biosensor based on magnetic aptamer for the visual and quantitative detection of ZEN. Specifically, we utilized the docking-aided rational tailoring (DART) strategy to analyze intermolecular force and interaction sites between zearalenone (ZEN) and the aptamer, and optimize the long-chain aptamer step by step to enhance the binding affinity by 3.4 times. The magnetic bead-modified aptamer underwent conformational changes when competing with complementary sequences to bind with ZEN. Then, the released complementary sequences will be amplified in template-free mode with the presence of the terminal deoxynucleotidyl transferase (TdT), and generating T-rich sequences as the core sequences for the luminescence of copper nanoclusters. The luminescence could be visualized and quantitatively detected through ultraviolet irradiation. The proposed label-free aptasensor exhibited high sensitivity and specificity, with a low limit of detection (LOD) of 0.1 ng/mL., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Zearalenone (ZEN), a nonsteroidal estrogenic mycotoxin, causes enormous economic losses in the food and feed industries. Simple, rapid, low-cost, and quantitative analysis of ZEN is particularly urgent in the fields of food safety and animal husbandry. Using the bioluminescent bacterium Photobacterium phosphoreum T3, we propose a bioluminescence inhibition assay to evaluate ZEN levels quickly. The limit of detection (LOD), limit of quantification (LOQ), and quantitative working range of this bioluminescence inhibition assay were 0.1 µg/mL, 5 µg/mL, and 5-100 µg/mL, respectively. The concentration-response curve of the bioluminescence inhibition rate and ZEN concentration was plotted within the range 5 to 100 μg/mL, as follows: y = 0.0069x 2 - 0.0190x + 7.9907 (R 2 = 0.9943, y is luminescence inhibition rate, x is ZEN concentration). First, we used the bioluminescence inhibition assay to detect the remaining ZEN in samples treated with purified lactonohydrolase ZHD101. The bioluminescence inhibition assay results showed a strong correlation with the HPLC analysis. Furthermore, we successfully evaluated the overall toxicity of samples treated with purified peroxidase Prx and H 2 O 2 using the P. phosphoreum T3 bioluminescence inhibition assay. The results indicate that the degradation products of ZEN created by purified peroxidase Prx and H 2 O 2 showed little toxicity to P. phosphoreum T3. In this study, a simple, rapid, and low-cost assay method of zearalenone by bioluminescent P. phosphoreum T3 was developed. The bioluminescence inhibition assay could be used to estimate the efficiency of enzymatic degradation of ZEN.

A self-reporting molecularly-imprinted electrochemical sensor is prepared for the detection of Zearalenone (ZEA). Firstly, the reduced graphene nanoribbons and reduced graphene oxide (rGNR-rGO) were simultaneously modified onto a glassy carbon electrode (GCE) to improve the sensor's sensitivity. After electrodepositing copper nanoparticles onto the rGNR-rGO/GCE, cyclic voltammetry scanning was performed in potassium ferrocyanide solution, and copper hexacyanoferrate (CuHCF) was deposited onto rGNR-rGO/GCE to further improve the sensor's sensitivity while giving it self-reporting capability. Then, molecularly-imprinted polymer films were prepared on the CuHCF/rGNR-rGO/GCE to ensure the selectivity of the sensor. It is found that the linear range of ZEA detection by the constructed sensor is 0.25-500 ng·mL -1 , with a detection limit of 0.09 ng·mL -1 . This sensor shows the merits of good selectivity, high sensitivity and accurate detection, providing a great possibility for the precise detection of low concentration ZEA in food., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

DON and ZEN residues in the blood and urine of dairy cows can be used to predict the outer exposure to DON and ZEN expressed per kilogram diet for a risk evaluation based on comparisons to critical dietary concentrations. This method was used to evaluate the exposure of dairy cows from 12 farms located in Brandenburg, Germany, fed rations with unknown DON and ZEN concentrations (N = 244). The corresponding diet concentrations predicted by different methods from analyzed blood and urine samples varied significantly amongst farms from 0 to 1.6 mg/kg for DON and 0 to 3.0 mg/kg for ZEN at a reference dry matter content of 88% but independently of lactational state (post-partum vs. early lactation). This significant variation was noticed below the critical dietary DON concentration of 5 mg/kg, while the ZEN concentration in one farm exceeded the critical ZEN level of 0.5 mg/kg markedly. Predicted DON concentrations of rations increased with the proportion of maize silage, while the high ZEN concentration found in one farm was most likely related to a higher proportion of sugar beet pulp supposedly highly contaminated by ZEN. Exceeding the critical dietary ZEN concentration and significant variations in DON contents below the critical level was not related to performance, reproductive performance, and health-related traits of cows. For a more consistent evaluation of possible associations between the inner exposure of cows to DON and ZEN, more frequent longitudinal observations of both mycotoxin residue levels and performance and health traits are required., (© 2024. The Author(s).)

Herein we developed a multicolor lateral flow immunoassay (LFIA) test strip for rapid and simultaneous quantitative detection of aflatoxin B1 (AFB1) and zearalenone (ZEN). Three differently colored aggregation-induced emission nanoparticles (AIENPs) were designed as LFIA signal tags, with red and green AIENPs for targeting AFB1 and ZEN at the test line, and yellow AIENPs for indicating the validity of the test strip at the control (C) line. After surface functionalization with antibodies, the developed AIENP-based multicolor LFIA allows simultaneous and accurate quantification of AFB1 and ZEN using an independent C-line assisted ratiometric signal output strategy. The detection limits of AFB 1 and ZEN were 6.12 and 26 pg/mL, respectively. The potential of this method for real-world applications was well demonstrated in corn and wheat. Overall, this multicolor LFIA shows great potential for field screening of multiple mycotoxins and can be extended to rapid and simultaneous monitoring of other small molecule targets., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Mycotoxins are secondary products produced primarily by fungi and are pathogens of animals and cereals, not only affecting agriculture and the food industry but also causing great economic losses. The development of rapid and sensitive methods for the detection of mycotoxins in food is of great significance for livelihood issues. This study employed an amino-functionalized zirconium luminescent metal-organic framework (LOF) (i.e., UiO-66-NH 2) . Click chemistry was utilized to assemble UiO-66-NH 2 in a controlled manner, generating LOF assemblies to serve as probes for fluorescence-linked immunoassays. The proposed fluoroimmunoassay method for Zearalenone (ZEN) and Fumonisin B1 (FB1) detection based on the UiO-66-NH 2 assembled probe (CLICK-FLISA) afforded a linear response range of 1-20 μmol/L for ZEN, 20 μmol/L for FB1, and a very low detection limit (0.048-0.065 μmol/L for ZEN; 0.048-0.065 μmol/L for FB1). These satisfying results demonstrate promising applications for on-site quick testing in practical sample analysis. Moreover, the amino functionalization may also serve as a modification strategy to design luminescent sensors for other food contaminants.

This study aimed to develop a novel fluorescent aptasensor for the quantitative detection of zearalenone (ZEN), addressing the limitations of conventional detection techniques in terms of speed, sensitivity, and ease of use. Nitrogen-doped carbon dots (N-CDs) were synthesized via the hydrothermal method, resulting in spherical particles with a diameter of 3.25 nm. These N-CDs demonstrated high water solubility and emitted a bright blue light at 440 nm when excited at 355 nm. The fluorescence of N-CDs was quenched by dispersed gold nanoparticles (AuNPs) through the inner filter effect, while aggregated AuNPs induced by NaCl did not affect the fluorescence of N-CDs. The aptamer could protect AuNPs from NaCl-induced aggregation, but the presence of ZEN weakened this protective effect. Based on this principle, optimal conditions for ZEN detection included 57 mM NaCl, 12.5 nM aptamer concentration, incubation of AuNPs with NaCl for 15 min in Tris-EDTA(TE) buffer, and incubation of aptamer with ZEN and NaCl for 30 min. Under these optimized conditions, the "signal-on" fluorescent aptasensor for ZEN detection showed a linear range of 0.25 to 200 ng/mL with a low detection limit of 0.0875 ng/mL. Furthermore, the developed aptasensor exhibited excellent specificity and could rapidly detect ZEN in corn flour samples or corn oil, achieving satisfactory recovery rates ranging from 84.7% to 108.6%. Therefore, this study presents an economical, convenient, sensitive, and rapid method for accurately quantifying ZEN in cereal products.

Herein, the aptamer-antibody sandwich module was first introduced to accurately recognize a low molecular weight compound (mycotoxin). Impressively, compared with the large steric hindrance of a traditional dual-antibody module, the aptamer-antibody sandwich with low Gibbs free energy and a low dissociation constant has high recognition efficiency; thus, it could reduce false positives and false negatives caused by a dual-antibody module. As a proof of concept, a sensitive electrochemiluminescence (ECL) biosensor was constructed for detecting mycotoxin zearalenone (ZEN) based on an aptamer-antibody sandwich as a biological recognition element and porous ZnO nanosheets (Zn NSs) supported Cu nanoclusters (Cu NCs) as the signal transduction element, in which the antibody was modified on the vertex of a tetrahedral DNA nanostructure (TDN) with a rigid structure to increase the kinetics of target recognition for promoting the detection sensitivity. Moreover, the Cu NCs/Zn NSs exhibited an excellent ECL response that was attributed to the aggregation-induced ECL enhancement through electrostatic interactions. The sensing platform achieved trace detection of ZEN with a low detection limit of 0.31 fg/mL, far beyond that of the enzyme-linked immunosorbent assay (ELISA, the current rapid detection method) and high-performance liquid chromatography (HPLC, the national standard detection method). The strategy has great application potential in food analysis, environmental monitoring, and clinical diagnosis.

Aflatoxin B1 (AFB1) and zearalenone (ZEN) are two mycotoxins that often co-occur in corn. A surface-enhanced Raman scattering-based lateral flow immunoassay (SERS-LFIA) that can simultaneously detect AFB1 and ZEN in corn samples was developed employing the core-interlayer-satellite magnetic nanocomposites (Fe 3 O 4 @PEI/Au MBA @Ag MBA ) as dual-functional SERS tags. Under the optimal conditions, the detection ranges of AFB1 and ZEN in corn samples were 0.1-10 μg/kg and 4-400 μg/kg, respectively. Moreover, the test results for two mycotoxins in contaminated corn samples employing the suggested SERS-LFIA was in line with those of the HPLC technique. In view of its satisfactory sensitivity, accuracy, precision and short testing time (20 min), the developed system has a promising application prospect in the on-site simultaneous detection of AFB1 and ZEN., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

It is challenging to prepare sample pretreatment materials with simple use, strong selectivity and satisfactory enrichment performance. In this study, the antibody (3D4) that can specifically recognize zearalenone (ZEN) and its metabolites was immobilized on the surface of gold-coated magnetic Fe 3 O 4 nanoparticles (GMN) by streptavidin (SA)-biotin interaction using GMN as the substrate and our designed four-arm PEG derivative (HS-4ARMPEG10K-(CM) 3 ) as the linker. The immunomagnetic nanoparticles (GMN-4ARMPEG10K-SA-3D4) prepared by this strategy can achieve rapid enrichment (only 5 min) of analytes directly in the matrix, and higher enrichment capacity compared with the previous immunomagnetic particles. The sensitive and accurate analysis of ZEN and its metabolites can be achieved coupled with HPLC-MS/MS. The LODs and LOQs were 0.02-0.05 μg/kg and 0.05-0.10 μg/kg, respectively. The recoveries were 84.13%-112.67%, and the RSDs were 1.09%-9.39%. The method can provide a powerful tool for highly sensitive and rapid monitoring of mycotoxins in complex matrices due to its' strong selectivity and resistance to matrix interference., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Ochratoxin A (OTA), zearalenone (ZEN), and deoxynivalenol (DON) are mycotoxins whose exposure is associated with various adverse health effects, including cancer and renal disorders, estrogenic effects, and immunosuppressive and gastrointestinal disorders, respectively. Infants (<2 years) are the most vulnerable group to mycotoxins, representing a unique combination of restricted food consumption types, low body weight, lower ability to eliminate toxins, and more future years to accumulate toxins. This study aimed to estimate the infant́s exposure to OTA, DON, and ZEN due to the consumption of milk formula and baby cereals in Chile. Milk formula samples (n = 41) and baby cereals (n = 30) were collected and analyzed using commercial ELISA kits for OTA, DON, and ZEA determination. Exposure was assessed by the Estimated Daily Intake (EDI) approach (mean and worst-case scenario, WCS) with the levels found in a modified Lower Bound (mLB) and Upper Bound (UB); ideal consumption (<6m, 7-12 m, and 13-24 m); adjusted by the weight of each group. The risk was estimated by comparing the EDI with a reference tolerable daily intake or by the margin of exposure (MOE) in the case of OTA. DON and OTA occurrence in infant formula were 34 % and 41 %, respectively. The co-occurrence between these mycotoxins was 22 %. Mycotoxin contents were below LOQ values except for OTA determined in one sample (0.29 ng/ml). No milk formulae were contaminated with ZEN. In the case of baby cereals, the occurrences were 17 % for OTA, 30 % for DON, and 7 % for ZEN, all below LOQ. Co-occurrence was seen in two samples between ZEN and OTA. According to exposure calculations, the MOE for OTA was less than 10,000 in all models for milk formula between 0 to 12 months of age and in the UB and WCS for cereal consumption. Health concerns were observed for DON in the WCS and UB for milk consumption in all ages and only in the UB WCS for cereal consumption. Considering the high consumption of milk formula in these age groups, regulation of OTA and other co-occurring mycotoxins in infant milk and food is strongly suggested., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin and seriously threatens food safety, which requires rapid and sensitive detection methods for monitoring ZEN in agro-products. Herein, an alkaline phosphatase-tagged single-chain variable fragment fusion protein (ALP-scFv) was used as a bifunctional tracer to develop a colorimetric enzyme immunoassay (CEIA) and a chemiluminescent enzyme immunoassay (CLEIA) for ZEN. In addition, the interactions between scFv and ZEN were exploited by computer-assisted simulation, and four key amino acid sites were preliminarily identified. After optimization, the CEIA and CLEIA exhibited a limit of detection of 0.02 and 0.006 ng/mL, respectively. Furthermore, both methods showed favorable accuracy in recovery experiments and good selectivity in cross reactions. Moreover, the detection results of the actual samples from both methods correlated well with those from high-performance liquid chromatography. Overall, the ALP-scFv fusion tracer-based CEIA and CLEIA are demonstrated as reliable tools for ZEN detection in food., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Efficient enrichment of trace zearalenone (ZEN) from the complex traditional Chinese medicine (TCM) samples is quite difficult, but of great significance for TCM quality control. Herein, we reported a novel magnetic solid phase extraction (MSPE) strategy for ZEN enrichment using the amino- and hydroxyl dual-functionalized magnetic microporous organic network (Fe 3 O 4 @MON-NH 2 -OH) as an advanced adsorbent combined with the high-performance liquid chromatography (HPLC) determination. Efficient extraction of ZEN was achieved via the possible hydrogen bonding, hydrophobic, and π-π interactions between Fe 3 O 4 @MON-NH 2 -OH and ZEN. The adsorption capacity of Fe 3 O 4 @MON-NH 2 -OH for ZEN was 215.0 mg g -1 at the room temperature, which was much higher than most of the reported adsorbents. Under the optimal condition, the developed Fe 3 O 4 @MON-NH 2 -OH-MSPE-HPLC method exhibited wide linear range (5-2500 μg L -1 ), low limits of detection (1.4-35 μg L -1 ), less adsorbent consumption (5 mg), and large enhancement factor (95) for ZEN. The proposed method was successfully applied to detect trace ZEN from 10 kinds of real TCM samples. Conclusively, this work demonstrates the Fe 3 O 4 @MON-NH 2 -OH can effectively extract trace ZEN from the complex TCM matrices, which may open up a new way for the application of MONs in the enrichment and extraction of trace contaminants or active constituents from the complex TCM samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Zearalenone (ZEN) is an extremely hazardous chemical widely existing in cereals, and its high-sensitivity detection possesses significant significance to human health. Here, the cathodic aggregation-induced electrochemiluminescence (AIECL) performance of tetraphenylethylene nanoaggregates (TPE NAs) was modulated by solvent regulation, based on which an electrochemiluminescence (ECL) aptasensor was constructed for sensitive detection of ZEN. The aggregation state and AIECL of TPE NAs were directly and simply controlled by adjusting the type of organic solvent and the fraction of water, which solved the current shortcomings of low strength and weak stability of the cathode ECL signal for TPE. Impressively, in a tetrahydrofuran-water mixed solution (volume ratio, 6:4), the relative ECL efficiency of TPE NAs reached 16.03%, which was 9.2 times that in pure water conditions, and the maximum ECL spectral wavelength was obviously red-shifted to 617 nm. In addition, "H"-shape DNA structure-mediated dual-catalyzed hairpin self-assembly (H-D-CHA) with higher efficiency by the synergistic effect between the two CHA reactions was utilized to construct a sensitive ECL aptasensor for ZEN analysis with a low detection limit of 0.362 fg/mL. In conclusion, solvent regulation was a simple and efficient method for improving the performance of AIECL materials, and the proposed ECL aptasensor had great potential for ZEN monitoring in food safety.

An electrochemical aptasensor was used for the fast and sensitive detection of zearalenone (ZEN) based on the combination of Co 3 O 4 /MoS 2 /Au nanocomposites and the hybrid chain reaction (HCR). The glassy carbon electrode was coated with Co 3 O 4 /MoS 2 /Au nanomaterials to immobilize the ZEN-cDNA that had been bound with ZEN-Apt by the principle of base complementary pairing. In the absence of ZEN, the HCR could not be triggered because the ZEN-cDNA could not be exposed. After ZEN was added to the surface of the electrode, a complex structure was produced on the modified electrode by the combination of ZEN and ZEN-Apt. Therefore, the ZEN-cDNA can raise the HCR to produce the long-strand dsDNA structure. Due to the formation of dsDNA, the methylene blue (MB) could be inserted into the superstructure of branched DNA and the peak currents of the MB redox signal dramatically increased. So the concentration of ZEN could be detected by the change of signal intensity. Under optimized conditions, the developed electrochemical biosensing strategy showed an outstanding linear detection range of 1.0×10 -10 mol/L to 1.0×10 -6 mol/L, a low detection limit (LOD) of 8.5×10 -11 mol/L with desirable selectivity and stability. Therefore, the fabricated platform possessed a great application potential in fields of food safety, medical detection, and drug analysis., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

With the aim of reintroducing wheat grains naturally contaminated with mycotoxins into the food value chain, a decontamination strategy was developed in this study. For this purpose, in a first step, the whole wheat kernels were pre-treated using cold needle perforation. The pore size was evaluated by scanning electron microscopy and the accessibility of enzymes and microorganisms determined using fluorescent markers in the size range of enzymes (5 nm) and microorganisms (10 μm), and fluorescent microscopy. The perforated wheat grains, as well as non-perforated grains as controls, were then incubated with selected microorganisms (Bacillus megaterium Myk145 and B. licheniformis MA572) or with the enzyme ZHD518. The two bacilli strains were not able to significantly reduce the amount of zearalenone (ZEA), neither in the perforated nor in the non-perforated wheat kernels in comparison with the controls. In contrast, the enzyme ZHD518 significantly reduced the initial concentration of ZEA in the perforated and non-perforated wheat kernels in comparison with controls. Moreover, in vitro incubation of ZHD518 with ZEA showed the presence of two non-estrogenic degradation products of ZEA: hydrolysed zearalenone (HZEA) and decarboxylated hydrolysed ZEA (DHZEA). In addition, the physical pre-treatment led to a reduction in detectable mycotoxin contents in a subset of samples. Overall, this study emphasizes the promising potential of combining physical pre-treatment approaches with biological decontamination solutions in order to address the associated problem of mycotoxin contamination and food waste reduction., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Mycotoxins are secondary metabolites of certain moulds, prevalent in 60-80% of food crops and many processed products but challenging to eliminate. Consuming mycotoxin-contaminated food and feed can lead to various adverse effects on humans and livestock. Therefore, testing mycotoxin residue levels is critical to ensure food safety. Gold standard analytical methods rely on liquid chromatography coupled with optical detectors or mass spectrometers, which are high-cost with limited capacity. This study reported the successful development of a microfluidic "lab-on-a-chip" device to enrich and detect zearalenone in food samples based on the fluorescence quenching effect of quantum dots and selective affinity of molecularly imprinted polymers (MIPs). The dummy template and functional polymer were synthesized and characterized, and the detailed microfluidic chip design and optimization of the flow conditions in the enrichment module were discussed. The device achieved an enrichment factor of 9.6 (±0.5) in 10 min to quantify zearalenone spiked in food with high recoveries (91-105%) at 1-10 mg kg -1 , covering the concerned residue levels in the regulations. Each sample-to-answer test took only 20 min, involving 3 min of manual operation and no advanced equipment. This microfluidic device was mostly reusable, with a replaceable detection module compatible with fluorescence measurement using a handheld fluorometer. To our best knowledge, the reported device was the first application of an MIP-based microfluidic sensor for detecting mycotoxin in real food samples, providing a novel, rapid, portable, and cost-effective tool for monitoring mycotoxin contamination for food safety and security.

Fusarium asiaticum is a predominant fungal pathogen causing Fusarium Head Blight (FHB) in wheat and barley in China and is associated with approximately £201 million in annual losses due to grains contaminated with mycotoxins. F. asiaticum produces deoxynivalenol and zearalenone whose maximum limits in cereals and cereals-derived products have been established in different countries including the EU. Few studies are available on the ecophysiological behaviour of this fungal pathogen, but nothing is known about the impact of projected climate change scenarios on its growth and mycotoxin production. Therefore, this study aimed to examine the interacting effect of i) current and increased temperature (25 vs 30 °C), ii) drought stress variation (0.98 vs 0.95 water activity; a w ) and iii) existing and predicted CO 2 concentrations (400 vs 1000 ppm) on fungal growth and mycotoxin production (type B trichothecenes and zearalenone) by three F. asiaticum strains (CH024b, 82, 0982) on a wheat-based matrix after 10 days of incubation. The results showed that, when exposed to increased CO 2 concentration (1000 ppm) there was a significant reduction of fungal growth compared to current concentration (400 ppm) both at 25 and 30 °C, especially at 0.95 a w . The multi-mycotoxin analysis performed by LC-MS/MS qTRAP showed a significant increase of deoxynivalenol and 15-acetyldeoxynivalenol production when the CH024b strain was exposed to elevated CO 2 compared to current CO 2 levels. Zearalenone production by the strain 0982 was significantly stimulated by mild water stress (0.95 a w ) and increased CO 2 concentration (1000 ppm) regardless of the temperature. Such results highlight that intraspecies variability exist among F. asiaticum strains with some mycotoxins likely to exceed current EU legislative limits under prospected climate change conditions., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

A certified reference material (CRM, KRISS 108-01-002) for zearalenone in corn flour was developed to assure reliable and accurate measurements in testing laboratories. Commercially available corn flour underwent freeze-drying, pulverization, sieving, and homogenization. The final product was packed in amber bottles, approximately 14 g per unit, and preserved at -70 °C. 13 C 18 -Zearalenone was used as an internal standard (IS) for the certification of zearalenone by isotope-dilution liquid chromatography-tandem mass spectrometry (ID-LC‒MS/MS) and for the analysis of α-zearalenol, β-zearalenol, and zearalanone by LC‒MS/MS. The prepared CRM was sufficiently homogeneous, as the among-unit relative standard deviation for each mycotoxin ranged from 2.2 to 5.7 %. Additionally, the stability of the mycotoxins in the CRM was evaluated under different temperature conditions and scheduled test periods, including storage at -70°C, -20°C, and 4°C and room temperature for up to 12 months, 6 months, and 1 month, respectively. The content of each target mycotoxin in the CRM remained stable throughout the monitoring period at each temperature. Zearalenone content (153.6 ± 8.0 µg/kg) was assigned as the certified value. Meanwhile, the contents of α-zearalenol (1.30 ± 0.17 µg/kg), β-zearalenol (4.75 ± 0.33 µg/kg), and zearalanone (2.09 ± 0.16 µg/kg) were provided as informative values., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)

The main mycotoxins involved in adverse equine health issues are aflatoxins, fumonisins, trichothecenes, and probably ergovaline (fescue grass endophyte toxicosis). Most exposures are through contaminated grains and grain byproducts, although grasses and hays can contain mycotoxins. Clinical signs are often nonspecific and include feed refusal, colic, diarrhea, and liver damage but can be dramatic with neurologic signs associated with equine leukoencephalomalacia and tremorgens. Specific antidotes for mycotoxicosis are rare, and treatment involves stopping the use of contaminated feed, switching to a "clean" feed source, and providing supportive care., Competing Interests: Disclosure The authors declare that there are no competing financial interests in the work described., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Contamination by toxic substances is a major global food safety issue, which poses a serious threat to human health. Mycotoxins are major class of food contaminants, mainly including aflatoxins (AFs), zearalenone (ZON), deoxynivalenol (DON), ochratoxin A (OTA), fumonisins (FBs) and patulin (PAT). Ferroptosis is a newly identified iron-dependent form of programmed or regulated cell death, which has been found to be involved in diverse pathological conditions. Recently, a growing body of evidence has shown that ferroptosis is implicated in the toxicities induced by certain types of food-borne mycotoxins, which provides novel mechanistic insights into mycotoxin-induced toxicities and paves the way for developing ferroptosis-based strategy to combat against toxicities of mycotoxins. In this review article, we summarize the key findings on the involvement of ferroptosis in mycotoxin-induced toxicities and propose issues that need to be addressed in future studies for better utilization of ferroptosis-based approach to manage the toxic effects of mycotoxin contamination., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

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.

Electrochemical aptasensors have emerged as promising platforms for effectivelydetection of various target analytes. Here, we developed a sensitive and selective electrochemical aptasensor for zearalenone (ZEN) determination based on a bimetallic organic framework (CuBi-BPDC). The results of HR-TEM, FE-SEM, XPS, etc. indicate the CuBi-BPDC possessing mixed nodes of Cu(II) and Bi(III) and multilayered nanosheets bearing nanoparticles. Due to its improved electrochemical activity and strong affinity for aptamers, the CuBi-BPDC-based aptasensor obtains a low limit of detection of 0.19 fg mL -1 (IUPAC S/N = 3) in a wide range of 1 fg mL -1 -10 ng mL -1 via EIS and 0.73 fg mL -1 from 0 fg mL -1 to 1 × 10 7 fg mL -1 via DPV for ZEN detection, respectively. Moreover, the excellent selectivity allows this aptasensor to specifically identify ZEN from other interfering substances in raw milk and rice, indicating the potential applicability of the CuBi-BPDC-based aptasensor in sensitive and selective detection of ZEN., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Exposure to mycotoxins through food is a major health concern, especially for youngsters. This study performed a preliminary investigation on children's exposure to dietary mycotoxins in Ribeirão Preto, Brazil. Sampling procedures were conducted between August and December 2022, to collect foods (N = 213) available for consumption in the households of children (N = 67), including preschoolers (aged 3-6 years, n = 21), schoolers (aged 7-10 years, n = 15), and adolescents (aged 11-17 years, n = 31) cared in the Vila Lobato Community Social Medical Center of Ribeirão Preto. Ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) was used to determine concentrations of the mycotoxins in foods. Mycotoxins measured in all foods comprised aflatoxins (AFs), fumonisins (FBs), zearalenone (ZEN), T-2 toxin, deoxynivalenol (DON) and ochratoxin A (OTA). Higher incidence and levels were found for FBs, ZEN, and DON in several commonly consumed foods. Furthermore, 32.86 % foods had two to four quantifiable mycotoxins in various combinations. The mean estimated daily intake (EDI) values were lower than the tolerable daily intake (TDI) for AFs, FBs, and ZEN, but higher than the TDI (1.0 µg/kg bw/day) for DON, hence indicating a health risk for all children age groups. Preschoolers and adolescents were exposed to DON through wheat products (EDIs: 2.696 ± 7.372 and 1.484 ± 2.395 µg/kg body weight (bw)/day, respectively), while schoolers were exposed through wheat products (EDI: 1.595 ± 1.748 µg/kg bw/day) and rice (EDI: 1.391 ± 1.876 µg/kg bw/day). The results indicate that wheat-based foods and rice may be risky to children, implying the need for stringent measures to avoid DON contamination in these products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: DNA tweezers, classified as DNA nanomachines, have gained prominence as multifunctional biosensors due to their advantages, including a straightforward structure, response mechanism, and high programmability. While the DNA tweezers demonstrate simultaneous, rapid, and stable responses to different targets, their detection sensitivity requires enhancement. Some small molecules, such as mycotoxins, often require more sensitive detection due to their extremely high toxicity. Therefore, more effective signal amplification strategies are needed to further enhance the sensitivity of DNA tweezers in biosensing., Results: We designed programmable DNA tweezers that detect small-molecule mycotoxins and miRNAs through simple sequence substitution. While the DNA tweezers demonstrate simultaneous, rapid, and stable responses to different targets, their detection sensitivity requires enhancement. We introduced the Strand Displacement Amplification (SDA) technique to address this limitation, proposing a strategy of novel programmable DNA tweezers-SDA ultrasensitive signal amplification fluorescence sensing. We specifically investigate the effectiveness of this approach concerning signal amplification for two critical mycotoxins: aflatoxin B1 (AFB1) and zearalenone (ZEN). Results indicate that the detection ranges of AFB1 and ZEN via this strategy were 1-10,000 pg mL -1 and 10-100,000 pg mL -1 , respectively, with corresponding detection limits of 0.933 pg mL -1 and 1.07 pg mL -1 . Compared with the DNA tweezers direct detection method for mycotoxins, the newly constructed programmable DNA tweezers-SDA fluorescence sensing strategy achieved a remarkable 10 4 -fold increase in the detection sensitivity for AFB1 and ZEN., Significance: The constructed programmable DNA tweezers-SDA ultrasensitive signal-amplified fluorescence sensing strategy exhibits excellent detection performance for mycotoxins. The superb versatility of this strategy allows the developed method to be easily used for detecting other analytes by simply replacing the aptamer and cDNA, which has incredible potential in various fields such as food safety screening, clinical diagnostics, and environmental analysis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Covalent organic frameworks (COFs) are attractive adsorbents for sample pretreatment due to their unique structure and properties. However, the selectivity of COFs for the extraction of hazardous compounds is still limited due to the lack of specific interactions between COFs and targets. Herein, we report a pore size adjustment strategy for room-temperature synthesis of molecularly imprinted COF (MICOF) for selective extraction of zearalenone (ZEN) in complex food samples. The three-dimensional building block tetra(4-aminophenyl) methane was used as a functional monomer, while dialdehyde monomers with different numbers of benzene ring were used to adjust the pore size of MICOF to match with the size of ZEN molecules. The prepared MICOF gave the largest adsorption capacity of 177.2 mg g -1 and the highest imprinting factor of 10.1 for ZEN so far. MICOF was used as the adsorbent for dispersed solid-phase extraction in combination with high-performance liquid chromatography for the determination of trace ZEN in cereals. The high selectivity of the developed method allows simple aqueous standard calibration for the matrix effect-free determination of ZEN in food samples. The limit of detection and the recoveries of the developed method were 0.21 μg kg -1 and 93.7-101.4%, respectively. The precision for the determination of ZEN was less than 3.8% (RSD, n = 6). The developed method is promising for the selective determination of ZEN in complex matrices.

In this work, a bimetallic organic framework (Cu/UiO-66) based "turn on" fluorescent aptamer probe was designed for the high-efficiency detection of zearalenone (ZEN). In the probe, the 6-carboxyfluorescein-labeled aptamer (FAM-Apt) was used as the recognition element, and the electrostatic interaction, coordination effect, and photoinduced electron transfer effect between FAM-Apt and Cu/UiO-66 caused fluorescence quenching. When ZEN existed, FAM-Apt recognized ZEN specifically, causing FAM-Apt to separate from the surface of Cu/UiO-66 and recovery of fluorescence. Under the optimal conditions, the probe had a linear detection range of 0.5 ng/mL-60 ng/mL, and the detection limit was 0.048 ng/mL. The application potential of the probe was verified by real detection of various cereals and their products, with a standard recovery from 83.67 %-106.8 %. The development of this efficient, rapid, and sensitive ZEN detection method provides a new platform for the quality control of cereals and their products., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Risk assessment primarily relies on toxicological data of individual substances, with limited information on combined effects. Recent in vitro experiments using Ishikawa cells, an endometrial carcinoma cell line expressing both estrogen receptor isoforms, demonstrated interactive effects of phyto- and mycoestrogens. The mycoestrogens, zearalenone (ZEN), and α-zearalenol (α-ZEL) exhibited significantly enhanced estrogenic responses in the presence of isoflavones (ISF), depending on substance ratios and concentrations. This study investigated the impact of phyto- and mycoestrogen combinations on estrogenic response following OECD guideline 455, utilizing hERα-HeLa-9903 cells. Test substances included mycoestrogens (ZEN and α-ZEL) and isoflavones (genistein (GEN), daidzein (DAI), and S-equol (EQ), a gut microbial metabolite of DAI). Mycoestrogens were tested in the range of 0.001 to 100 nM, while isoflavones were used at concentrations 1000 times higher based on relevant occurrence ratios. Results showed that ZEN and α-ZEL induced ERα-dependent luciferase expression in concentrations above 1 nM and 0.01 nM, respectively. However, ISF caused a superinduction of the luciferase signal above 1 µM. A superinduction is characterized by an unusually strong or heightened increase in the activity of the luciferase enzyme. This signal is not affected by the estrogen receptor antagonist 4-hydroxytamoxifen (4-OH-TAM), which was additionally used to verify whether the increase of signal is a true reflection of receptor activation. This superinduction was observed in all combinations of ZEN and α-ZEL with ISFs. Contrary to the luciferase activity findings, RT-qPCR experiments and a stability approach revealed lower real ERα activation by ISFs than measured in the ONE-Glo™ luciferase test system. In conclusion, the OECD protocol 455 appears unsuitable for testing ISFs due to their superinduction of luciferase and interactions with the test system, resulting in experimental artifacts. Further studies are necessary to explore structure-activity relationships within polyphenols and clarify the test system's applicability., (© 2023. The Author(s).)

Rapid, sensitive, specific and stable detection of mycotoxin in food remains an extremely crucial issue. Herein, a magnetic-fluorescent immunosensing platform for the detection of zearalenone (ZEN) was proposed. The platform utilized Au nanoparticles (AuNPs) heterogeneous fluorescent metal-organic framework (MIL-53(Al)@AuNPs) labeled with ZEN-bovine serum albumin (ZEN-BSA) as signal probe and ZEN mono-antibodies coupled with magnetic NPs (MNPs-mAbs) as capture probe. Specifically, the heterogenization of AuNPs on the MIL-53(Al) surface improved its biocompatibility, and facilitated the loading of ZEN-BSA conjugates. The MNPs-mAbs could rapidly capture the target ZEN, simplify the immunoassay process and further improve the detection efficiency. The established competitive magnetic-fluorescent immunosensing platform had a wider linear response to ZEN in the range of 0.001-100 ng/mL with a lower limit of detection (LOD) at 0.0035 ng/mL, and could finish the whole detection process within 20 min, showing great potential for rapid and sensitive detection of food contaminants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

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 (B 2 -ssDNA). The enzyme nanocomposites improved the loading amount of ALP and catalyzed more ascorbic acid 2-phosphate to generate ascorbic acid (AA). Subsequently, Cu 2+ 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., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)