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A GPE-PET (graphene-polyethylene terephthalate) bipolar electrode-electrochemiluminescence (BPE-ECL) platform was developed for ochratoxin A (OTA) detection. PET served as the electrode sheet substrate, and GPE was drop-coated onto the surface of PET to form a conductive line. On the functional sensing interface, the thiol (-SH) modified OTA aptamer (OTA-Aptamer) are fixed on the surface of the gold-plated cathode through AuS bonds. The efficient electron transfer ability of methylene blue (MB) made the anode ECL signal strong. Due to competition between OTA and MB with OTA-Aptamer, leading to a decrease in ECL intensity of the [Ru(bpy) 3 ] 2+ /TPA system on the BPE anode. Under optimized conditions, the GPE-PET BPE-ECL biosensor displayed superior sensitivity for OTA with a detection limit of 2 ng mL -1 and a wide linear concentration range of 5-100 ng mL -1 . This method could be further applied to detect various toxins and had broad application prospects., 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.)

Aspergillus westerdijkiae can grow as 'golden mould' on the surface of dry-cured meat products, causing deterioration, and can produce the mycotoxin known as ochratoxin A that is a risk to human health. The aim of this study was to evaluate the growth and ochratoxin production by A. westerdijkiae at different temperatures (15, 20, and 25 °C), in salami-based culture media supplemented with glycerol and NaCl to adjust the water activity (0.85, 0.90, 0.93, 0.97 and 0.99). The growth of the two strains of A. westerdijkiae (S1 and S2) was evaluated for 28 days and, after this period, ochratoxin A was extracted from the culture media and analysed by high-performance liquid chromatography. For both strains, the optimal growth (>10 mm day -1 ) occurred at 0.97 and 0.99 water activity at 20-25 °C. The lowest water activity (0.85) and temperature (15 °C) resulted in the slowest growth rates (<2.0 mm day -1 ). However, ochratoxin A production by strain S1 was highest (21 μg g -1 ) at 20 °C on glycerol-supplemented media at 0.97 water activity; the strain S2 produced its higher level of ochratoxin A (7.0 μg g -1 ) at water activity 0.99 and 20 °C on NaCl-supplemented media. Whereas some microorganisms produce toxic secondary metabolites under stress/boundary conditions, we noted that mycotoxin production by A. westerdijkiae occurred at optimal growth conditions (high water activities). These conditions also show a more intense interspecies competition of microbial communities. Insights into the ecophysiology of fungi can be used to make knowledge-based decisions to reduce contamination of dry-cured meat products., Competing Interests: Declaration of competing interest I wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome. I confirm that the manuscript has been approved by all named authors and that there are no other persons who satisfied the criteria for authorship but are not listed. I understand that the Corresponding Author is the sole contact for the Editorial process (including Editorial Manager and direct communications with the office)., (Copyright © 2024 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Human breast milk is the optimal source of nutrition for newborns, but the potential transfer of contaminants like mycotoxins, particularly ochratoxin A (OTA), from maternal blood to milk remains a concern. This systematic review aims to provide a comprehensive analysis of global OTA levels in human breast milk and assess the associated health risks. We conducted a thorough search of scientific databases, including Web of Science, ScienceDirect, Scopus, Google Scholar and PubMed, using keywords related to OTA in human breast milk. A total of 39 studies met the inclusion criteria for this review. OTA levels compared to limits, estimated infant intake at various ages and health risks assessed using Margin of Exposures (MOEs) and Hazard quotient (HQ). Our findings reveal the widespread presence of OTA in breast milk across different regions, with notably higher levels detected in Africa compared to Asia, South America and Europe. The higher concentrations observed in warmer, humid climates suggest that environmental factors significantly influence OTA contamination. Mature breast milk samples generally exhibited greater OTA exposure. The neoplastic and non-neoplastic effects demonstrate generally low risks globally. The regional differences in OTA levels and associated health risk assessments underscore the need for continued research into the health impacts of OTA exposure in infants. This includes further investigation into multiple sources of exposure, such as infant formula, within the broader context of the exposome framework.

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

Coffee is one of the most commonly consumed beverages worldwide, so assessing its quality for potential health risks is essential. Therefore, this review aimed to determine the levels of ochratoxin A (OTA) in coffee worldwide and then estimate its human intake and health risks. The systematic search took place from June 1997 to April 2024 and 40 of 254 articles were selected based on the selection criteria. The results showed significant differences in average levels of OTA between countries, continents and coffee types ( p  < 0.001). Of 3256 samples, OTA was detected in 1778, accounting for 54.6% of the total, with the percentage of positive results varying between 7.5% and 100%. Only two studies reported OTA average levels in roasted coffee exceeding the maximum limit (ML) set by the European Commission (ML-EC = 5 μg/kg). The average OTA in soluble coffee was lower than ML-EC (10 μg/kg) in all studies, and in instant coffee, the level of OTA was higher than ML-EC (10 μg/kg) only in one study. The estimated daily intake (EDI) of OTA in all coffee types was lower than the provisional tolerable daily intake (PTDI) values set by joint FAO/WHO Expert Committee on Food Additives (JECFA) (14 ng/kg bw/day) and proposed by the European Food Safety Authority (EFSA) (17 ng/kg bw/day). Non-carcinogenic risk assessment through coffee consumption indicated that the hazard quotient (HQ) was below the acceptable level, HQ = 1. The Margin of Exposure (MoE) for neoplastic effects was acceptable and unacceptable for non-neoplastic effects (NNE) in 4.5% (one of 22 cases) of the roasted and soluble coffees, but acceptable for all instant coffees. In conclusion, the study shows that the OTA content of coffee is not toxic to consumers worldwide. However, preventative measures should be taken, including inhibiting fungal growth and reducing OTA-producing fungal growth.

Co-contamination of multiple mycotoxins produces synergistic toxic effects, leading to more serious hazards. Therefore, the simple, rapid and accurate simultaneous detection of multiple mycotoxins is crucial. Herein, a three-channel aptamer-based lateral flow assay (Apt-LFA) was established for the detection of aflatoxin M 1 (AFM 1 ), aflatoxin B 1 (AFB 1 ) and ochratoxin A (OTA). The multi-channel Apt-LFA utilized gold‑iridium nanozyme to catalyze the chromogenic substrate, which effectively achieved signal amplification. Moreover, the positions and lengths of the complementary sequences were screened by changes in fluorescence intensity. After grayscale analysis, the semi-quantitative results showed that the detection limits of AFM 1 , AFB 1 and OTA were 0.39 ng/mL, 0.36 ng/mL and 0.82 ng/mL. The recoveries of the multiplexed competitive sensors in complex matrices of real samples were 93.33%-97.01%, 95.72%-102.67%, and 106.88%-109.33%, respectively. In conclusion, the assembly principle of the three-channel Apt-LFA is simple, which can provide a new idea for the simultaneous detection of small molecule targets., 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.)

Ochratoxin A (OTA) is a nephrotoxin that contaminates grains in storage. Moisture and temperature sensors give delayed responses due to their slow kinetic movement within the silo. This study examines if CO 2 production could predict OTA contamination and identify storage conditions exceeding the maximum limit (5 μg/kg). The impact of water activity levels (0.70-0.90 a w ), temperatures (15 and 20 °C), and storage duration on (a)Penicillium verrucosum population, (b)CO 2 respiration rates (RR), and (c)ochratoxins concentrations in stored wheat was investigated. 96 samples were analysed for ochratoxins with LCMS-MS. RR was >7 times higher at wetter conditions than at drier a w levels. A positive correlation between CO 2 , OTA, OTB, and OTα was observed at the wettest conditions. OTA exceeded the limit at >0.80 a w (16% moisture content) with RR > 0.01 mg CO 2 kg -1  h -1 . The knowledge of the RR of stored grain would alert grain farmers/managers to improve grain storage management., 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 Authors. Published by Elsevier Ltd.. All rights reserved.)

Ginger, a fresh rhizome, an economically important spice with extensive nutraceutical activities finds itself in vegetable and therapeutic market. Aflatoxins (AFB 1 , AFB 2 , AFG 1 and AFG 2 ) along with ochratoxin A (OTA) are the most significant and the most toxic form of mycotoxins which are produced by various fungi. This study was initiated to assess the contamination of AFs and OTA in raw and dried ginger products, collected from different agro-climatic zones in Punjab, Pakistan employing the high performance liquid chromatography. We found all (raw ginger samples commercial ginger powders) samples contaminated with AFB 1 (range: 29.88-1060.12 μg/kg). AFB 2 contamination was much lower (range: 0-17.54 μg/kg). Variable contamination of AFG 1 was also observed (range: 0-170.58 μg/kg) whereas AFG 2 contamination was found in only three (range: 0-21.88 μg/kg) out of 19 raw ginger samples. OTA contamination ranged from 0.05 to 3.42 μg/kg. Ginger samples from lower altitudes (<1000 m) were more contaminated with AFB 1 sub type mycotoxin. Keeping in view that the toxicity of AFs is in the order of B 1 >G 1 > B 2 >G 2 , it was alarming to find that 100% of the samples were contaminated with AFB 1 way beyond the permissible limits. Our very first report about the contamination of ginger with AFs presents a grave health issue because of wide use of ginger. We conclude that ginger production in Pakistan needs to be carefully crafted and due diligence is needed during ginger cultivation, harvest and post-harvest operations because the amount of aflatoxins detected in this study are very much above the permissible limits. In this regard, ginger storage in cooler environments such as refrigerator should be encouraged to contain the AFs proliferation., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Ochratoxin A (OTA) is a group of mycotoxins that are widely distributed in food and feed and are closely associated with human health, so it is particularly important to detect OTA in cereal-based foods. Porphyrins and their derivatives have been widely investigated for their excellent electrochemical luminescence properties. Tetrakis 4-carboxyphenyl) porphyrin (TCPP) has limited applications because of its tendency to aggregate in water., Results: To enhance the luminescence efficiency of TCPP, the porphyrin can be immobilized as an organic ligand in a metal-organic framework. This allows the preparation of a novel zinc-porphyrin-based MOF (Zn-TCPP nanorods), which in turn provides highly efficient and stable cathodic ECL signals. Herein, an ultra-sensitive electrochemiluminescence immunoassay was proposed using Zn-TCPP nanorods as high-efficiency luminophores and Bi 2 S 3 @Au nanoflowers as electrode substrate materials for the detection of ochratoxin A in foodstuffs. Zn-TCPP has a strong and stable signal, and has been used as an immunosensor probe material. The Bi 2 S 3 @Au nanoflowers was used to decorate the glass carbon electrode and support for antibody immobilization due to its good electrical conductivity and large specific surface area. Under the optimized conditions, the constructed immunosensor could realize the sensitive detection of ochratoxin A in the detection range of 0.0004 ng mL -1 to 500 ng mL -1 with the detection limit as low as 0.13 pg mL -1 . In addition, the sensing platform has been used for the detection of OTA in wheat flour and feed., Significance: Hence, it is worth believing that this strategy can pave a bright research direction for the detection of ochratoxin A or other small molecule mycotoxins content in foods, as well as contributing to the further study of MOF in the field of ECL., Competing Interests: Declaration of competing interest All the authors listed have approved the submission: The first author Xiaolin Fan declares that she has no conflict of interest. The second author Cheng Tan declares that he has no conflict of interest. The third author Xiao Mei declares that he has no conflict of interest. The fourth author Jun Ma declares that she has no conflict of interest. The fifth author Kang Wu declares that he has no conflict of interest. The sixth author Anping Deng declares that he has no conflict of interest. The seventh author Xinjian Feng declares that he has no conflict of interest. Corresponding author Jianguo Li declares that he has no conflict of interest. Informed consent was obtained from all individual participants included in the study., (Copyright © 2024. Published by Elsevier B.V.)

Freekeh is produced from roasted, immature wheat grains. It is very popular in Middle Eastern and North African nations. This study aimed to evaluate the occurrence of different types of mycotoxins, physical impurities, and microbiological contamination in local freekeh products. Lateral flow competitive immunochromatographic assay was used to evaluate the occurrence of mycotoxins. It was found that physical impurities for some tested products exceeded the permitted limit (>2% of straw and foreign grains). Moreover, our findings showed that total aerobic bacterial and fungal counts in Freekeh products varied from 1 to 4 logs and from 1.39 to 4.3 logs, respectively. The incidence ranges of aflatoxins and ochratoxin were 3.17-3.33 ppb and 4.63-8.17 ppb, respectively. The levels of deoxynivalenol (DON) and T2/HT2 (trichothecene T2 and deacetylated form HT2) were less than the limit of detection. More than 78% of Freekeh samples tested had aflatoxin and ochratoxin contents higher than the limit permitted by the European Commission (4 and 5 ppb). In conclusion, gaining knowledge about the quality, safety, and labeling of freekeh products can help increase their commercial potential. Further investigations are needed to evaluate the factors affecting contamination levels within the freekeh supply chain.

Ochratoxin A (OTA) is a mycotoxin produced by fungi species belonging to the genera Aspergillus spp. and Penicillium spp. The proliferation of OTA-producing fungal species may occur due to inadequate practices during both the pre-harvest and post-harvest stages of feed. Consequently, poultry species may be exposed to high concentrations of this mycotoxin that can be transferred to animal tissues due to its carry-over, reaching dangerous concentrations in meat and meat products. Therefore, this review aims to propose a comprehensive overview of the effects of OTA on human health, along with data from global studies on the prevalence and concentrations of this mycotoxin in avian feeds, as well as in poultry meat, edible offal, and eggs. Moreover, the review examines significant gross and histopathological lesions in the kidneys and livers of poultry linked to OTA exposure. Finally, the key methods for OTA prevention and decontamination of feed are described.

Throughout history, Turkish coffee has been the most widely consumed type of coffee in Turkey. Therefore, the goal of this study was to determine the amount of ochratoxin A (OTA) present in Turkish coffee and to analyze any potential health hazards. A total of 41 Turkish coffees were collected and analyzed for OTA activity using a competitive enzyme immunoassay (ELISA). Furthermore, dietary exposure and health risk assessments for the Turkish population were calculated based on analytical results and coffee consumption data from the Turkish Nutrition and Health Survey 2019 (TNHS-2019) in three age categories (15-18, 19-64, and 65 + years). Nine of the samples contained more than 2.5 μg/kg OTA, with an average of 5.24 μg/kg. The OTA concentration in 3 samples exceeded the permissible maximum limit (5 μg/kg) established by Turkish legislation, and the mean concentration was 8.41 μg/kg. A margin of exposure (MOE) approach was used for risk characterization, considering both non-neoplastic and neoplastic consequences. There were no concerns about health risks because MOEs were more than 10,000 for all categories. Although the levels of OTA analyzed in Turkish coffee did not pose a risk to individuals in the three age categories, emphasis should be placed on minimizing and controlling OTA concentrations in Turkish coffee. Additionally, it is also necessary to consider other food sources that could contribute to OTA exposure., (© 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.)

Mycotoxins, such as aflatoxin B 1 (AFB 1 ), deoxynivalenol (DON), fumonisins (FBs), ochratoxin A (OTA), T-2 toxin (T-2), and zearalenone (ZEN), can contaminate animal feeds and pose risks to animal health and production performance. These mycotoxins are commonly found in cereals and grains, with the increased use of cereals in pet food, there is a rising concern about mycotoxin contamination among pet owners. To address this, we analyzed imported brands of feline and canine food from the Chinese market produced in 2021-2022. Ninety-three samples were analyzed, comprising 45 feline food and 48 canine food samples. Among them, 14 were canned food and 79 were dry food. The results indicate that AFB 1 , DON, FBs, OTA, T-2, and ZEN occurred in 32.26%, 98.92%, 22.58%, 73.12%, 55.91%, and 7.53% of the samples, respectively. The most prevalent mycotoxin was DON, followed by OTA, T-2, AFB 1, and FBs, whereas ZEN was less frequently detected. The mean concentrations of the six mycotoxins in pet feed samples were 3.17 μg/kg for AFB 1 , 0.65 mg/kg for DON, 2.15 mg/kg for FBs, 6.27 μg/kg for OTA, 20.00 μg/kg for T-2, and 30.00 μg/kg for ZEN. The levels of mycotoxins were generally below the limits of the Pet Feed Hygiene Regulations of China and the EU. Notably, a substantial majority of the pet food samples (88 out of 93) were contaminated by two or more mycotoxins. AFB 1 , FBs, OTA, and ZEN occurred slightly more often in feline food than in canine food. Except for OTA, the contamination rates for the other five mycotoxins in canned food were lower than those in dry food. Moreover, except for AFB 1 , the levels of the other five mycotoxins in canned foods were lower than those in dry foods. This study highlights the widespread contamination of pet foods with mycotoxins, which poses a significant risk to pets from continuous exposure to multiple mycotoxins., (© 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.)

An innovative aptasensor incorporating MoS 2 -modified bicolor quantum dots and a portable spectrometer, designed for the simultaneous detection of ochratoxin A (OTA) and aflatoxin B1 (AFB1) in corn was developed. Carbon dots and CdZnTe quantum dots were as nano-donors to label OTA and AFB1 aptamers, respectively. These labeled aptamers were subsequently attached to MoS 2 receptors, enabling fluorescence resonance energy transfer (FRET). With targets, the labeled aptamers detached from the nano-donors, thereby disrupting the FRET process and resulting in fluorescence recovery. Furthermore, a portable dual-mode fluorescence detection system, complemented with customized python-based analysis software, was developed to facilitate rapid and convenient detection using this dual-color FRET aptasensor. The developed host program is connected to the spectrometer and transmits data to the cloud, enabling the device to have Internet of Things (IoT) characteristics. Connected to the cloud, this IoT-enabled device offers convenient and reliable fungal toxin detection for food safety., 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.)

The European Union legislation regarding ochratoxin A (OTA) in various foodstuffs has changed relatively recently. Nevertheless, the legislation does not regulate OTA in any meat and meat-derived products. In this legislation update, the European Commission requested new studies, including, besides others, the presence of OTA in hams, which raises the concern that its consumption may pose a potential risk of exposure to OTA. This study aims to investigate OTA in a total of 195 samples of air-dry-cured hams acquired at the Czech market from January to June 2023. The analytical technique of high-performance liquid chromatography in combination with a fluorescence detector with pre-treatment employing immunoaffinity columns was used to determine OTA. OTA was found in 93 (48%) samples of air-dry-cured ham, with the OTA concentration reaching up to 14.58 ng/g. Due to the current absence of regulation limits, the results of this study were compared with the Italian maximum limit of 1 ng/g regulating OTA in porcine meat and byproducts. The Italian OTA maximum limit was exceeded in 22 (11%) samples. This study shows that the population of the Czech Republic is exposed to OTA from this pork byproduct. It is essential to set an OTA regulatory limit for meat and food produced from it to protect human health., 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.)

The occurrence of Ochratoxin A (OTA) in wine is commonly known, but there is only limited information about its occurrence in grape mash and wines of German origin. Climate change has led to higher temperatures in the southern regions of Germany, which may increase the growth of fungi associated with the production of OTA and increase the content of this mycotoxin in grapes. A safe and rapid UHPLC-FLD method was developed and validated to assess the contamination of grape mash and wine with OTA. A total of 71 samples of grape mash and 30 wines from various wine producers in Baden-Württemberg, Germany, were analysed for OTA content. The results showed that no samples contained OTA in concentrations above the limit of detection. Further monitoring of samples from different vintages is needed., (© 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.)

Developing an efficient method for screening Ochratoxin A (OTA) in agriculture products is vital to ensure food safety and human health. However, the complex food matrix seriously affects the sensitivity and accuracy. To address this issue, we designed a novel molecularly imprinted polymer (MIP) electrochemical sensor based on multiwalled carbon nanotube-modified niobium carbide (Nb 2 C-MWCNTs) with the aid of the density functional theory (DFT). In this design, a glassy carbon electrode (GCE) was first modified by Nb 2 C-MWCNTs heterostructure. Afterward, the MIP layer was prepared, with ortho-toluidine as a functional monomer selected via DFT and OTA acting as a template on the surface of Nb 2 C-MWCNTs/GCE using in-situ electropolymerization. Electrochemical tests and physical characterization revealed that Nb 2 C-MWCNTs improved the sensor's active surface area and electron transmission capacity. Nb 2 C-MWCNTs had a good synergistic effect on MIP, endowing the sensor with high sensitivity and specific recognition of OTA in complex food matrix systems. The MIP sensor showed a wide linear range from 0.04 to 10.0 μM with a limit of detection (LOD) of 3.6 nM. Moreover, it presented good repeatability and stability for its highly antifouling effect on OTA. In real sample analysis, the recoveries, ranging from 89.77% to 103.70%, agreed well with the results obtained by HPLC methods, suggesting the sensor has good accuracy and high potential in practical applications., Competing Interests: Declaration of competing Interest The authors have no conflicts of interest to declare that are relevant to the content of this article., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Limitations in the sensitivity, linear detection range, and cross-reaction of lateral flow immunoassays mainly hamper their application in rapid screening for multiple targets. In this work, we designed a new time-resolved fluorescence immunoassay (TRFIA) platform to overcome these limitations. This platform uses europium chelate polystyrene (PS@Eu) nanoparticles conjugated with monoclonal antibodies to sense multiple mycotoxins. We employed a competitive TRFIA protocol in which the conjugated PS@Eu was used on the surfaces of photonic microbead arrays (PMAs). The TRFIA signal of PMAs on the pad was recorded with the digital time-resolved fluorescence reader. The developed TRFIA shows wide detection linear ranges (0.01-1000 ng/mL for DON, 0.1-100 ng/mL for OTA, and 0.01-100 ng/mL for AFB 1 ), low limits of detection (LODs) (7.9 pg/mL for DON, 18 pg/mL for OTA, and 7.7 pg/mL for AFB 1 ), good specificity, good recovery ratios (76.68-117.26%), and good reproducibility in grain samples. The simulated fluorescence enhancement effect of PMA indicated that the electric field distribution on the surface of PS@Eu on PMA is twice higher than that on the surface of PS@Eu. The new TRFIA for three kinds of mycotoxins was 1000-fold more sensitive than the classical TRFIA, and it has great potential application in rapid screening for multiple targets.

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.

The high toxicity and widespread contamination of ochratoxin A (OTA) make it urgent to develop a sensitive method to detect trace OTA in complex food matrices. Herein, an indirect competitive enzyme-linked immunosorbent assay (icELISA)-based on the CRISPR/Cas12a system is described. DNA amplicons with multiple activation sequences of the CRISPR/Cas12a system were pre-prepared to improve detection sensitivity. In the absence of OTA, streptavidin-mediated biotinylated DNA amplicons were captured by the biotinylated secondary antibody on the microplate. The captured DNA amplicons activated the CRISPR/Cas12a system, which thereby effectively cleaved the reporter DNA, producing strong fluorescence. The presence of OTA led to a decrease in DNA amplicons on the microplate, resulting in a decrease in activated Cas12a and ultimately a drop in fluorescence intensity. OTA in food matrices at nanogram per milliliter levels can be detected. Therefore, the new method has great potential in monitoring OTA.

Background: Ultrasensitive detection is crucial for the early warning and intervention of risk factors, ultimately benefiting the environment and human health. Low levels of ochratoxin A (OTA) present a hidden yet significant threat, and rapid detection via high-performing biosensors is therefore essential., Results: A cascade isothermal amplification aptasensor (CIA-aptasensor) was designed for OTA detection. On the surface of a magnetic bead probe, the OTA level was converted into positively correlated trigger cDNA through its competitive binding with OTA-Apt. The released trigger cDNA activated catalytic hairpin assembly followed by coupling with a hybridization chain reaction to achieve CIA. After adding graphene oxide and SYBR Green I, the background interference was eliminated to specifically obtain OTA-related fluorescence. The ultrasensitive limit of detection was 0.22 pg mL -1 , an improvement of 1368-fold over conventional enzyme-linked aptamer sorbent assay by the same OTA-Apt, demonstrating satisfactory reliability and practicability. Thus, the CIA-aptasensor provides an enzyme- and label-free simplified homogeneous system with minimal background interference using isothermal conditions., Significance: This study provides a polymerase chain reaction-like approach for enhancing the sensitivity and performance of a biosensor, which could be extended for the application of CIA and label-free signaling strategy to other risk factors., 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.)

This study investigated the role of irradiation with a high-energy electron beam in reducing mycotoxin levels of African nutmeg powder (ANP) samples. African nutmeg was procured from a local market in Accra, Ghana, cleaned, milled, packaged and irradiated using electrons of energy 9 MeV at doses of 2, 4, 6 and 8 kGy. Un-irradiated ANP served as a control. Mycotoxin levels of the treated samples were determined using appropriate standard methods. Aflatoxins B1 (AFB1) and B2 (AFB2) as well as ochratoxin A (OTA) were detected in the nutmeg samples. Irradiation significantly ( p  < 0.05) reduced mycotoxin levels of the ANP with increasing doses. Aflatoxins G1 and G2 were not detected in any of the samples. A dose of 8 kGy was effective in reducing the mycotoxin levels below the permissible limit in food. This suggests that a high-energy electron beam is effective in reducing mycotoxin levels in African nutmeg powder.

In response to the pressing need for highly efficient simultaneous detection of multiple mycotoxins, which are often found co-occurring in food raw materials and feed, an MXene-based electrochemical aptasensor array (MBEAA) was developed. This aptasensor array utilizes high-specificity aptamers as recognition elements, enabling the capture of electrical signal changes in the presence of target mycotoxins. Based on this platform, a multi-channel portable electrochemical device, enabling rapid, cost-effective, and simultaneous detection of aflatoxin B1 (AFB1), ochratoxin A (OTA), and zealenone (ZEN) was further developed. The developed system boasts a wide detection range of 1.0 × 10 -1 to 10.0 ng mL -1 , with remarkable performance characterized by ultra-low detection limits of 41.2 pg mL -1 , 27.6 pg mL -1 , and 33.0 pg mL -1 for AFB1, OTA, and ZEN, respectively. Successfully applied in corn samples, this method offers a portable, easy-to-operate, and cost-effective solution for simultaneous multi-mycotoxin detection. Moreover, the application of the self-developed detection system could be expanded for simultaneous detection of many different targets when their specific aptamers or antibodies were available., 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.)

Penicillium nordicum is the main ochratoxin A (OTA)-producing species on the surface of dry-fermented sausages, such as the "chorizo". New antifungal strategies are being developed using biocontrol agents (BCAs), such as plant extracts and native microorganisms. This work aimed to evaluate the antiochratoxigenic capacity and the causative modes of action of BCAs (rosemary essential oil (REO), acorn shell extract and the yeast Debaryomyces hansenii (Dh)) in a "chorizo"-based medium (Ch-DS). BCAs were inoculated on Ch-DS together with P. nordicum and incubated at 12 °C for 15 days to collect mycelia for OTA analyses and comparative proteomics. Both REO and Dh alone decreased OTA accumulation up to 99% and affected the abundance of P. nordicum proteins linked to cell wall organisation, synthesis of OTA-related metabolites and ergosterol synthesis. It is worth highlighting the increased abundance of an amidase by REO, matching with the decrease in OTA. The use of REO and Dh as BCAs could be an effective strategy to reduce the OTA hazard in the meat industry. Based on their not fully coincident modes of action, their combined application could be of interest in "chorizo" to maximise their potential against ochratoxigenic strains., 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.)

Timely and accurate on-site detection of ochratoxin A (OTA) is extremely important for global public health. In this study, a fluorescence/colorimetric biosensor based on Ti 3 C 2 nano-materials (Ti 3 C 2 -NMS) and a machine-learning (ML) based fluorescence/colorimetric intelligent learning system for detection of OTA concentration (COTA) were developed. The sensor was fabricated by functionalizing Ti 3 C 2 -NMS prepared by physical-exfoliation assisted metal-ion-induction using ssDNA. The Ti 3 C 2 -NMS exhibited good fluorescence quenching characteristics (FQC) and peroxidase-like activity (PLA). More surprisingly, the functionalization of Ti 3 C 2 -NMS by ssDNA further enhanced the FQC and PLA of the material, which could be used for dual-mode detection of OTA. When different COTA existed, ssDNA competitively bound to OTA, resulting in regular changes in fluorescence and colorimetric signals of the sensor, which realized the accurate and sensitive biosensing detection of OTA in two modalities. Based on a series of fluorescent/colorimetric RGB datasets collected by a self-developed application, a dual-channel ML model had been developed. This model can be integrated into mobile phones, clouds, and PCs to achieve intelligent sensing detection of OTA with the assistance of fully connected artificial neural networks. The method constructed had high specificity, low cost, and fast responsiveness, with a LOD as low as 1.58 pg mL -1 , indicating excellent potential for application and promotion., 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 B.V.)

Established chromatographic techniques for mycotoxin control in foodstuffs require prior sample enrichment and clean-up, typically achieved using immunoaffinity columns (IACs). Bovine serum albumin (BSA) has recently emerged as a cost-effective alternative to antibodies used in IACs. This study aimed at exploring the BSA domain II (BDII), which houses the primary binding site for ochratoxin A (OTA), as a bioreceptor for OTA capture. Recombinant BDII (rBDII) was produced in soluble form by Escherichia coli Origami 2(DE3), fused to a His6 (HisBDII) or thioredoxin-His6 (TrxBDII) tag, with yields up to 19 ± 4.3 mg/L culture in shake-flask. Fluorescence and circular dichroism (CD) spectroscopy revealed interaction of OTA with both rBDII variants, with estimated binding constants for OTA-HisBDII/TrxBDII complexes in the range of 5.7-9.3 × 10 4  M -1 . CD also showed an α/β structure of rBDII variants, in opposition to the predominant α-helical structure of whole BSA, and slight increase in their α-helical content upon binding to OTA. TrxBDII immobilized on Ni-NTA resin successfully captured OTA from spiked samples at the optimum pH range of 6.5-7.0, allowing OTA extraction, clean-up, and enrichment from spiked white grape juice, with up to 84 ± 7.4 % recovery., 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 Authors. Published by Elsevier B.V. All rights reserved.)

Mycotoxins are widely prevalent in various agricultural commodities, whose excessive consumption can pose significant risks to human health. In this study, we developed a facile mycotoxin detection platform based on branched hybridization chain reaction coupled with lateral flow assay. Ochratoxin A/Aflatoxin B1 bind to aptamers triggering the release of initiators, which leads to bHCR amplification and forms three-dimensional dendritic DNA nanostructures. Using the functionalized quantum dots as a fluorescent label, by leveraging smartphones and handheld ultraviolet lamps, the qualitative and quantitative detection of OTA and AFB1 can be achieved with a significantly enhanced sensitivity level, surpassing that of commercial test strips by 2-3 orders of magnitude. The visual detection limits for OTA and AFB1 were 30 pg/mL and 4 pg/mL, respectively. This approach eliminates the necessity for enzyme catalysis or the preparation and purification of antibodies and/or hapten, thereby reducing testing expenses and streamlining operational procedures. Moreover, substituting aptamer and nucleic acid sequences can effectively expand the scope of detection targets. Consequently, the as-proposed strategy exhibits great potential as a versatile technique, suitable for various analytical scenarios 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.)

Ochratoxin A (OTA) poses significant risks to human health, being potentially nephrotoxic, carcinogenic, genotoxic, and immuno-toxic. In this work, we developed a dual-mode aptasensor for OTA analysis, integrating colorimetric, electrochemical, and smartphone-based detection. The bifunctional Fe-MIL-88 metal-organic framework, acting as both a nanozyme capable of catalyzing the 3,3',5,5'-tetramethylbenzidine substrate and an electrochemical signal amplifier, enabled OTA quantification through current response or changes in color and absorbance intensity. Besides, the spatial confinement effect enhances the local concentration of Fe-MIL-88 signal probes through rolling circle amplification reaction, thereby contributing to a substantial enhancement in sensitivity. The proposed technique is simple, disposable, highly sensitive and selective, enabling OTA detection in the range of 1 fg/mL to 250 ng/mL, with a limit of detection of 0.22 fg/mL (3σ rule). Furthermore, we successfully detected OTA in corn, wheat, and red wine samples, with results good concordance with those obtained using commercial enzyme-linked immunoassay kits., 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.)

Multiple and sensitive mycotoxin detection is an essential early-warning mechanism for safeguarding human health, and preserving the environment. We synthesized a turn-on Fluorescence Resonance Energy Transfer (FRET) aptamer sensor based on the unique fluorescence quenching and substrate recognition characteristics of Ag NTs (energy receptors) and aptamer modified Fe 3 O 4 @TiO 2 NP (energy donor) to detect multiple toxins using a single diagnostic approach. The addition of aflatoxin B1 (AFB1) and ochratoxin A (OTA) resulted in a change in fluorescence intensity at 510 and 650 nm, which can be employed for simultaneous recognition with detection limits of 0.94 ng·mL -1 (R 2  = 0.997) and 0.54 ng·mL -1 (R 2  = 0.995). The aptasensors have been successfully applied for the determination of AFB1 and OTA in grain and oil samples with high recovery rates. The approach provides novel possibilities for the development of sensitive and selective aptasensors with potential applications in aptamer-recognized multifunctional biosensing., 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.)

In this work, a porphyrin metalation-based ratiometric absorbance and fluorescence dual model immunoassay was proposed to detect ochratoxin A (OTA). 5,10,15,20-tetrakis(1-methyl-4-pyridinio) porphyrin (TMPyP) was pink and had a strong fluorescence, upon coordination with Hg(II), its fluorescence was quenched and the color became green. The alkaline phosphatase can catalyze the dephosphorylation of ascorbic acid 2-phosphate to produce ascorbic acid, which can reduce the coordinated Hg(II) to Hg(0) and then dissociated from TMPyP, its fluorescence was recovered. Meanwhile, the color changed from green to light pink, which can be identified by naked eye for semi-quantitative detection. The linear ranges of ratiometric absorbance and fluorescence model were 0-6.0 ng/mL and 0.1-6.0 ng/mL, respectively. The absorbance and fluorescence signals produced by porphyrin metalation can mutually verify to improve the accuracy of detection results. Besides, the ultra-sensitivity and high selectivity demonstrated this method was a powerful tool for trace OTA detection., 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: Ochratoxin A (OTA) is a serious food contaminant, not easily degradable, and capable of causing irreversible damage to the human body. Therefore, it is of great practical significance to establish a sensitive and efficient OTA detection method. The electrochemical aptasensor has a broad development prospect in OTA detection with its advantages of fast response speed and low cost., Results: Herein, a cascade signal amplification strategy based on AuPt NPs/Zr-MOF and π-structure bipedal DNA walker-triggered hybridization chain reaction (HCR) was designed for the detection of ochratoxin A (OTA). AuPt NPs/Zr-MOF was employed as the electrode modification material, providing a large number of active sites and high conductivity, achieving 1.47 times signal amplification. Interestingly, bipedal DNA walker binds to hairpin 1 (H1) to form the π-structure. Under the activation of Pb 2+ , one bipedal DNA walker can simultaneously bind and cleave two H1. It exhibits a wide walking range and high recognition efficiency. After H1 is cleaved, the trigger sequence was exposed to trigger HCR and a large amount of methylene blue was loaded on the electrode. Under the optimal conditions, the linear range of the determined OTA is 1 × 10 -3 -500 ng/mL, and the limit of detection is as low as 0.525 pg/mL., Significance: The experimental results demonstrate that the constructed electrochemical aptasensor is a sensitive and efficient platform for OTA monitoring. The applicability in food samples was also confirmed, and the strategy was efficiently selective and reproducible for different analytes. This provides ideas for subsequent food safety testing., 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.)

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.)

Photoelectrochemical (PEC) is a highly sensitive and fast analytical method that can be used at low concentrations, while photoelectrochromic is a simple and low-cost method primarily utilized for high concentration detection. Therefore, we have developed a dual-signal output sensing platform based on both PEC and photoelectrochromism for rapid and sensitive OTA detection. The sensing platform is divided into signal generation (SG) region and signal output (SO) region, which modified with WO 3 /BiVO 4 photoactive nanocomposites and polyaniline (PANI), respectively. By irradiating the SG region, photogenerated electrons are generated and injected into the SO region through the conductive pathway, resulting in a decrease in surface blue polyaniline and a change to green. The smart device can accurately measure the RGB-Green values, enabling the construction of a photochromic visual sensing platform. After immobilizing the OTA aptamer in the SG region, a linear correlation was observed between the concentration of OTA and the RGB-Green value within the range of 20 ng/L ∼250 μg/L. The detection limit was determined to be 8.33 ng/L (S/N = 3). Furthermore, for a more sensitive OTA detection, a PEC sensing platform was developed utilizing the SG region as a photoanode, exhibiting a linear correlation in the range of 2 pg/L∼300 μg/L with a detection limit of 0.8 pg/L (S/N = 3). The detection of these two modes under the requirement of the international standard for the maximum limit realizes the sensitive OTA detection. The RGB-Green is verified to PEC signal, which improves the detection accuracy. The sensing platform has several advantages and is suitable for various application scenarios., 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 novel Electrochemiluminescence Immunosensor was constructed using PCN-224-Mn and gold-platinum nanoflowers (AuPt NFs) for the ultrasensitive detection of ochratoxin A (OTA). PCN-224 modified with Mn (II) was synthesized as a probe material. The interaction efficiency of PCN-224 with S 2 O 8 2- was also greatly improved. AuPt NFs were used as the substrate material for the electrodes. It has favorable biocompatibility, large specific surface area and can bind more antigen. Also greatly increased the electroactive surface area and conductivity of the electrode. OTA was detected using a competitive immunoassay strategy, in which OTA in the sample competes with the encapsulated antigen for a finite number of antibodies. ECLIA for the detection of OTA was designed to be highly sensitive, with a linear range from 0.0002 ng mL -1 to 1000 ng mL -1 and a LOD as low as 0.067 pg mL -1 . In addition, it was evident from the electrochemical analyses that PCN-224-Mn had a stronger and more stable ECL signal compared to the plain PCN-224. The successful preparation of specific, sensitive and reproducible ECL immunosensors confirms the great promise for the detection of OTA or other small molecule mycotoxins., Competing Interests: Declaration of competing interest We wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome. We confirm that the manuscript has been read and approved by all named authors and that there are no other persons who satisfied the criteria for authorship but are not listed. We further confirm that the order of authors listed in the manuscript has been approved by all of us. We confirm that we have given due consideration to the protection of intellectual property associated with this work and that there are no impediments to publication, including the timing of publication, with respect to intellectual property. In so doing we confirmed that we have followed the regulations of our institutions concerning intellectual property. We further confirm that any aspect of the work covered in this manuscript that has involved experimental animals has been conducted with the ethical approval of all relevant bodies and that such approvals are acknowledged within the manuscript. We understand that the Corresponding Author is the sole contact for the Editorial process. He is responsible for communicating with the other authors about progress, submissions of revisions and final approval of proofs. We confirm that we have provided a current, correct email address which is accessible by the Corresponding Author and which has been configured to accept email from lijgsd@suda.edu.cn., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed to assess the presence of ochratoxin A (OTA) residues in the blood serum of slaughtered pigs in Greece. Samples were obtained from 1695 healthy slaughtered pigs originating from 113 different farms located in 21 geographic regional units in 8 different geographic regions of Greece and were analyzed using an immunosorbent assay (ELISA) and high-performance liquid chromatography with fluorescence detector (HPLC-FD). OTA contamination assessment showed that 782 (46.1%) and 1233 (72.7%) samples were OTA-positive, with a concentration range of 0.20-5.38 μg/L and 0.15-5.96 µg/L according to ELISA and HPLC-FD analysis, respectively. Also, 88 (77.9%) and 108 (95.6%) of farms were found to be OTA-positive by ELISA and HPLC-FD analysis, respectively. The highest OTA serum positivity rate (>98%) and toxin level (5.96 µg/L) determined by HPLC-FD were observed in the Thessaly region, whereas a high prevalence of up to 100% (range 75-100%) was found on farms in the Crete Island region. The detection of OTA in the serum of slaughtered pigs in different regions in Greece poses a risk for animal and human health and highlights the need for constant OTA monitoring in the swine industry and pork meat production facilities.

The food contamination with Ochratoxin A (OTA) has highlighted the need to create precise, sensitive, and convenient techniques. Herein, we proposed a label-free and immobilization-free ratiometric homogeneous electrochemical aptasensor based on dual catalytic hairpin self-assembly (CHA) for OTA detection. Methylene blue (MB) and ferrocene (Fc) in solution were utilized as label-free signaling molecules, generating a response signal (I MB ) and a reference signal (I Fc ), respectively. The ratio of I MB /I Fc was utilized as a measure to quantify OTA. Dual CHA was exploited to increase the ratiometric signal and enhance the amplification efficiency. This aptasensor achieved trace-level detection for OTA over a linear range of lower concentrations (1.0 × 10 -3 ng/mL-1.0 × 10 3 ng/mL) with LOD of 92 fg/mL. The aptasensor was successfully applied to detect OTA in cereal and wine, with comparable results of HPLC-MS/MS. This strategy provided a viable platform for rapid, sensitive, and accurate detection of OTA 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.)

Ochratoxin A (OTA) in food poses a serious challenge to public health. Herein, using the nanobody-driven controllable aggregation of gold nanoparticles (AuNPs) in a glucose oxidase-tyramine-horseradish peroxidase (GOx-TYR-HRP) system, we propose a direct competitive plasmonic enzyme immunoassay (dc-PEIA) for OTA detection. The OTA-GOx conjugate catalyzes glucose to produce hydrogen peroxide (H 2 O 2 ), and then HRP catalyzes H 2 O 2 to generate hydroxyl radical which induces the crosslink of TYR. Crosslinked TYR leads to aggregation of AuNPs through strong electrostatic interactions, which is tunable based on the competition of OTA-GOx and free OTA for binding the immobilized nanobody. The optimized dc-PEIA achieves an instrumental limit of detection (LOD) of 0.275 ng/mL and a visual LOD of 1.56 ng/mL. It exhibits good selectivity for OTA and accuracy in the analysis of pepper samples, with the confirmation of high-performance liquid chromatography. Overall, the dc-PEIA is demonstrated as a useful tool for detecting OTA 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.)

A multiplex assay of mycotoxins in food and medicine is urgently needed and challenging due to synergistic hazards of trace mycotoxins and a lack of sensitive and user-friendly detection approaches. Herein, a cobalt DNA-inorganic hybrid superstructure (Co@DS) was developed through isothermal rolling circle amplification (RCA) for an ultrasensitive chemiluminescence (CL) imaging assay of multiple mycotoxins. Cobalt ions were enriched in the RCA product, endowing the Co@DS with a high CL catalytic property. Experimental studies elucidated the formation and CL catalytic mechanism of Co@DS. Co@DS was facilely integrated with biotinylated DNA to function as a universal platform and combined with a disposable immunosensor array chip. After a competitive immunoassay and biotin-avidin recognition, the CL signals of luminol and hydrogen peroxide, catalyzed by Co@DS captured on each testing zone of the array chip, were imaged simultaneously. Target mycotoxins can be quantitated by CL intensities. To validate the concept, the CL imaging approach was employed for joint determination of aflatoxin B 1 , ochratoxins A, and zearalenone. Under optimal conditions, it showed advantages including simple sample pretreatment, acceptable throughput, high accuracy, minimal sample consumption, broad linear ranges, and detection limits as low as 0.75, 0.62, and 0.61 pg mL -1 , respectively. Furthermore, the approach was applied in analyzing real coix seed samples, showcasing excellent performance in effectively distinguishing qualified and contaminated medicine, revealing the great potential in managing the complex issue of mycotoxins cocontamination in food and medicine.

Nontoxic substitutes to mycotoxins can facilitate the development of eco-friendly immunoassays. To explore a novel nontoxic substitute to ochratoxin A (OTA), this study screened shark anti-idiotypic variable new antigen receptors (VNARs) against the alpaca anti-OTA nanobody Nb28 through phage display. After four rounds of biopanning of a naïve VNAR phage display library derived from six adult Chiloscyllium plagiosum sharks, one positive clone, namely, P-3, was validated through a phage enzyme-linked immunosorbent assay (phage ELISA). The recombinant anti-idiotypic VNAR AId-V3 was obtained by prokaryotic expression, and the interactions between Nb28 and AId-V3 were investigated via computer-assisted simulation. The affinity of AId-V3 for Nb28 and its heptamer Nb28-C4bpα was measured using Biacore assay. Combining Nb28-C4bpα with AId-V3, a novel direct competitive ELISA (dcELISA) was developed for OTA analysis, with a limit of detection of 0.44 ng/mL and a linear range of 1.77-32.25 ng/mL. The good selectivity, reliability, and precision of dcELISA were confirmed via cross-reaction analysis and recovery experiments. Seven commercial pepper powder samples were tested using dcELISA and validated using high-performance liquid chromatography. Overall, the shark anti-idiotypic VNAR was demonstrated as a promising nontoxic substitute to OTA, and the proposed method was confirmed as a reliable tool for detecting OTA 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 B.V. All rights reserved.)

In light of the significant risks that mycotoxins posed to public health and environmental safety, this research developed an adsorbent MIPs/Apt/AuNPs@ZIF-67 (MA-AZ) utilizing a dual-recognition approach combining molecularly imprinted polymers (MIPs) and aptamer (Apt). This innovative method enabled the effective and highly selective recognition and enrichment of ochratoxin A (OTA). ZIF-67 was utilized as a carrier with a substantial specific surface area, and gold nanoparticles (AuNPs) were loaded on its surface to fix the thiol-modified Apt on the surface of the carrier. Then, an initiator was used to initiate a polymerization reaction, and the generated MIPs coated Apt/AuNPs@ZIF-67, thereby synthesizing the MA-AZ with a "synergistic recognition" effect. The Apt significantly increased the number of recognition sites within the imprinted cavities, and MIPs played roles in identifying targets, fixing and protecting Apt. The combination of the both produced the effect of "1+1>2". The study on the adsorption performance of MA-AZ found that the adsorption capacity of MA-AZ could reach 65.1 mg/g, and the imprinted factor was 5.48. In addition, MA-AZ exhibited excellent stability, specificity, reusability and recovery rate. Thus, this study offers valuable insights for the recognition and enrichment of hazardous substances, and helps to promote the rapid development of safety detection., 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 materials for sample pretreatment due to their tunable structures and functions. However, the precise recognition of contaminant in complex environmental matrices by COFs remains challenging owing to their insufficient specific active sites. Herein, we report Co 2+ coordination-assisted molecularly imprinted flexible COF (MI-COF@Co 2+ ) for selective recognition of ochratoxin A (OTA). The MI-COF@Co 2+ was prepared via one-step polymerization of 3,3-dihydroxybenzidine, 2,4,6-tris(4-formylphenoxy)- 1,3,5-triazine, Co 2+ and template. The flexible units endowed COFs with the self-adaptable ability to regulate the molecular conformation and coordinate with Co 2+ to locate the imprinted cavities. The coordination interaction greatly improved the adsorption capacity and selectivity of MI-COF@Co 2+ for OTA. The prepared MI-COF@Co 2+ was used as solid phase extraction adsorbent for high-performance liquid chromatography determination of OTA with the detection limit of 0.03 ng mL -1 and the relative standard deviation of < 2.5 %. In addition, this method permitted interference-free determination of OTA in real samples with recovery from 89.5 % to 102.8 %. This work provides a simple way to improve the selectivity of COFs for the determination of hazardous compounds in complex environments., 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) is a mycotoxin that globally contaminates fruits and their products. Since OTA have a huge negative impact on health hazards and economic losses, it is imperative to establish an effective and safe strategy for detoxification. Here, pancreatin was immobilized on the surface of polydopamine functionalized magnetic porous chitosan (MPCTS@ PDA) for the degradation of OTA. Compared with free pancreatin, MPCTS@ PDA@ pancreatin displayed excellent thermal stability, acid resistance, storage stability and OTA detoxification in wine (>58%). Moreover, the MPCTS@ PDA@ pancreatin retained 43% initial activity after 8 reuse cycles. There was no significant change in the quality of wine after MPCTS@ PDA@ pancreatin treatment. Moreover, it did not exhibit cytotoxicity which facilitated its application in wine. These results demonstrated that MPCTS@ PDA@ pancreatin can be used as a highly effective biocatalysate for OTA detoxification in wine., 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.)

Here, we report a monomer planarity modulation strategy for room-temperature constructing molecularly imprinted-covalent organic frameworks (MI-COFs) for selective extraction of ochratoxin A (OTA). 2,4,6-triformylphloroglucinol (Tp) was used as basic building block, while three amino monomers with different planarity were employed as modulators to explore the effect of planarity on the selectivity of MI-COFs. The MI-TpTapa constructed from Tp and the lowest planarity of monomer Tapa gave the highest selectivity for OTA, and was further used as the adsorbent for dispersed-solid phase extraction (DSPE) of OTA in alcohol samples. Coupling MI-TpTapa based DSPE with high-performance liquid chromatography allowed the matrix-effect free determination of OTA in alcohol samples with the limit of detection of 0.023 μg kg -1 and the recoveries of 91.4-97.6%. The relative standard deviation (RSD, n = 6) of intra and inter day was <3.2%. This work provides a new way to construct MI-COFs for selective extraction of hazardous targets., 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. Published by Elsevier Ltd.)

Rapid, portable, and accurate detection tools for monitoring ochratoxin A (OTA) in food are essential for the guarantee of food safety and human health. Herein, as a proof-of-concept, this study proposed a ratiometric bioluminescence immunosensor (RBL-immunosensor) for homogeneous detection of OTA in pepper. The construct of the RBL-immunosensor consists of three components, including the large fragment of the split nanoluciferase (NanoLuc)-tagged nanobody (NLg), the small fragment of the split NanoLuc-tagged mimotope peptide heptamer (MPSm), and the calibrator luciferase (GeNL). The specific nanobody-mimotope peptide interaction between NLg and MPSm induces the reconstitution of the NanoLuc, which catalyzes the Nano-Glo substrate and produces a blue emission peak at 458 nm. Meanwhile, GeNL can produce a green emission peak at 518 nm upon substrate conversion via bioluminescent resonance energy transfer (BRET). Therefore, the concentration of OTA can be linked to the variation of the bioluminescence signal (λ 458 /λ 518 ) measured by microplate reader and the variation of the blue/green ratio measured by smartphone via the competitive immunoreaction where OTA competes with MPSm to bind NLg. The immunosensor is ready-to-use and works by simply mixing the components in a one-step incubation of 10 min for readout. It has a limit of detection (LOD) of 0.98 ng/mL by a microplate reader and an LOD of 1.89 ng/mL by a smartphone. Good selectivity and accuracy were confirmed for the immunosensor by cross-reaction analysis and recovery experiments. The contents of OTA in 10 commercial pepper powder samples were tested by the RBL-immunosensor and validated by high-performance liquid chromatography. Hence, the ready-to-use RBL-immunosensor was demonstrated as a highly reliable tool for detection of OTA 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 B.V. All rights reserved.)

This study reviews global levels of ochratoxin A (OTA) in infant formula and cereal-based foods, using Monte Carlo simulation to assess risks. The review found 24 studies on global OTA levels in infant food and cereal-based products, using databases including PubMed, Scopus, Web of Science and Embase until March 2024. We estimated OTA exposure in infant food based on concentration, intake and body weight. The exposure and hazard quotient margin were calculated using BMDL10 and TDI values. Monte Carlo simulation evaluated human health risks from OTA in infant formula and cereal-based foods. A global study from 14 countries shows varying levels, surpassing EU limits in Tunisia, Ecuador, the USA, and generally in Africa, notably in infant cereals, which had higher levels than formula. Globally, OTA was present in 29.3% of the 3348 samples analyzed, with Lebanon at 95.2% and Brazil at 0%. Analysis indicates only non-carcinogenic risk for infants. While health risks for infants are mostly low, ongoing research and monitoring are vital to minimize OTA exposure in infant food.

The production of raisins, a method of grape preservation since antiquity, has evolved with various drying techniques that significantly impact the quality and safety of the final product. This study evaluates the efficacy of a solar indirect dryer compared to traditional sun-drying methods for drying Centennial Seedless and Sultanina grape cultivars in Crete, Greece. Key parameters assessed include environmental conditions, drying time, grape color, fungal contamination, and Ochratoxin A (OTA) levels. Grapes were dried in a controlled solar chamber and under open sun conditions. The solar chamber maintained higher average temperatures (34 °C) and lower relative humidity (39.7%) than outside conditions (24.2 °C and 58.7%, respectively), significantly reducing the drying time from 12 to 7 days. Raisins dried in the solar chamber exhibited improved color quality, with higher Lightness (L*), Hue Angle (h), and Chroma (C*) values, attributed to minimized enzymatic and nonenzymatic browning. Mycological analysis revealed a substantial reduction in Aspergillus section Nigri contamination in chamber-dried raisins, with mean colony-forming units per gram significantly lower than those of sun-dried raisins. Consequently, OTA levels were also significantly reduced in chamber-dried raisins, with Centennial Seedless showing a mean concentration of 1.01 µg/kg compared to 2.66 µg/kg in sun-dried samples, and Sultanina showing 0.70 µg/kg versus 2.05 µg/kg, respectively. These findings underscore the advantages of using solar indirect dryers to enhance drying efficiency, improve color quality, and reduce fungal and OTA contamination, highlighting the importance of adopting controlled drying technologies for safer, higher-quality raisins., 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 Inc. All rights reserved.)

Background: The importance of multi-target simultaneous detection lies in its ability to significantly boost detection efficiency, making it invaluable for rapid and cost-effective testing. Photoelectrochemical (PEC) sensors have emerged as promising candidates for detecting harmful substances and biomarkers, attributable to their unparalleled sensitivity, minimal background signal, cost-effectiveness, equipment simplicity, and outstanding repeatability. However, designing an effective multi-target detection strategy remains a challenging task in the PEC sensing field. Consequently, there is a pressing need to address the development of PEC sensors capable of simultaneously detecting multiple targets., Results: CdIn 2 S 4 /V-MoS 2 heterojunctions were successfully prepared via a hydrothermal method. These heterojunctions exhibited a high photocurrent intensity, representing a 1.53-fold enhancement compared to CdIn 2 S 4 alone. Next, we designed a multi-channel aptasensing chip using ITO as the substrate. Three working electrodes were created via laser etching and subsequently modified with CdIn 2 S 4 /V-MoS 2 heterojunctions. Thiolated aptamers were then self-assembled onto the CdIn 2 S 4 /V-MoS 2 heterojunctions via covalent bonds, serving as recognition tool. By empolying the CdIn 2 S 4 /V-MoS 2 heterojunctions as the sensing platform and aptamers as recognition tool, we successfully developed a disposable aptasensing chip for the simultaneous PEC detection of three typical mycotoxins (aflatoxin B1 (AFB1), ochratoxin A (OTA), and zearalenone (ZEN)). This aptasensing chip exhibited wide detection range for AFB1 (0.05-50 ng/mL), OTA (0.05-500 ng/mL), and ZEN (0.1-250 ng/mL). Furthermore, it demonstrated ultra-low detection limits of 0.017 ng/mL for AFB1, 0.016 ng/mL for OTA, and 0.033 ng/mL for ZEN., Significance and Novelty: The aptasensing chip stands out for its cost-effectiveness, simplicity of fabrication, and multi-channel capabilities. The versatility and practicality enable it to serve as a powerful platform for designing multi-channel PEC aptasensors. With its ability to detect multiple targets with high sensitivity and specificity, the aptasensing chip holds immense potential for applications across diverse fields, such as environmental monitoring, clinical diagnostics, and food safety monitoring, where multi-target detection is crucial., 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.)

The study aimed to screen fungal diversity and ochratoxin A levels on culinary spice and herb samples sold in open-air markets and supermarkets in Nairobi County, Kenya. All herbs were grown in Kenya, while locally-produced and imported spices were purchased from both types of retail outlet. The results showed a high frequency of Aspergillus and Penicillium species contaminating the samples. The isolated species included Aspergillus ochraceous , Aspergillus nomiae , Aspergillus niger , Aspergillus flavus , Aspergillus ustus , Aspergillus terrus , Aspergillus nidulans , Aspergillus clavutus , Penicillium crustosum , Penicillium expansum , Penicillium brevicompactum , Penicillium glabrum , Penicillium thomii , Penicillium citrinum , Penicillium polonicum , and Cladosporium cladosporioides. Total fungal count on spice and herb samples collected from various sources varied between 6 and 7 CFU/mL. Of imported spices, garlic had the highest fungal diversity, while cardamom had the least. For spices from both open market and supermarket outlets, cloves had the highest fungal diversity, while white pepper had the least. For the herbs sampled from the open markets, basil was the most contaminated, while sage was the least. In supermarket samples, parsley, sage, and mint had the highest fungal diversity, and bay had the least. The results indicate the contamination of spices and herbs with OTA at high concentrations. The calibration curve was saturated at 40 µg/kg; with samples of garlic, cinnamon, red chili, basil, thyme, mint, sage, and parsley having levels above this. Of the spices, imported ginger had the highest OTA levels (28.7 µg/kg), while turmeric from the open market had the least, 2.14 µg/kg. For herb samples, parsley from the open market had the highest OTA levels at 29.4 µg/kg, while marjoram from the open market had the lowest at 6.35 µg/kg. The results demonstrate the presence of mycotoxigenic fungi and OTA contamination of marketed culinary herbs and spices beyond acceptable limits. Hence, there is a need for informed and sustainable mitigation strategies aimed at reducing human exposure in Kenya to OTA mycotoxicosis through dietary intake of spices and herbs.

Among cereal contaminants, mycotoxins are of concern due to their importance in terms of food and feed safety. The difficulty in establishing a diagnosis for mycotoxicosis relies on the fact that the effects are most often subclinical for chronic exposure and the most common scenario is multi-contamination by various toxins. Mycotoxin co-occurrence is a major food safety concern as additive or even synergic toxic impacts may occur, but also regarding current regulations as they mainly concern individual mycotoxin levels in specific foods and feed in the food chain. However, due to the large number of possible mycotoxin combinations, there is still limited knowledge on co-exposure toxicity data, which depends on several parameters. In this context, this systematic review aims to provide an overview of the toxic effects of two regulated mycotoxins, namely ochratoxin A and fumonisin B1. This review focused on the 2012-2022 period and analysed the occurrence in Europe of the selected mycotoxins in different food matrices (cereals and cereal-derived products), and their toxic impact, alone or in combination, on in vitro intestinal and hepatic human cells. To better understand and evaluate the associated risks, further research is needed using new approach methodologies (NAM), such as in vitro 3D models. KEY CONTRIBUTION: Cereals and their derived products are the most important food source for humans and feed for animals worldwide. This manuscript is a state of the art review of the literature over the last ten years on ochratoxin A and fumonisin B1 mycotoxins in these products in Europe as well as their toxicological effects, alone and in combination, on human cells. Future perspectives and some challenges regarding the assessment of toxicological effects of mycotoxins are also discussed., 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 Authors. Published by Elsevier B.V. All rights reserved.)

Developing sensitive and accurate Ochratoxin A (OTA) detection methods is essential for food safety. Herein, a simple and reliable strategy for regulating interenzyme distance based on a rigid DNA quadrangular prism as a scaffold was proposed to establish a new electrochemical biosensor for ultrasensitive detection of OTA. The interenzyme distances were precisely adjusted by changing the sequences of the hybridized portions of hairpins SH1 and SH2 to the DNA quadrangular prism, avoiding the complexity and instability of the previous DNA scaffold-based enzyme spacing adjustment strategies. The electrochemical biosensor constructed at the optimal interenzyme distance (10.4 nm) achieved sensitive detection of OTA in a dynamic concentration range from 10 fg/mL to 250 ng/mL with a detection limit of 3.1 fg/mL. In addition, the biosensor was applied to quantify OTA in real samples, exhibiting great application potential in food safety., 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.)