Multiple Mycotoxins Research Articles

Co-Occurrence of Mycotoxins in the Diet and in the Milk of Dairy Cows from the Southeast Region of Brazil

Mycotoxins are toxic fungi secondary metabolites that develop on feedstuffs and can be transferred into milk, thus representing a public health risk. The objective of this study was to assess the co-occurrence of mycotoxins in the diet and in the milk of dairy cows from the southeast region of Brazil. Samples of total mixed ration (TMR, n = 70) and milk (n = 70) were collected in dairy farms and subjected to multi-mycotoxin analysis using liquid chromatography coupled to tandem mass spectrometry. The aflatoxins (AFs), ochratoxin A (OTA), and T-2 and HT-2 toxins were not detected in TMR samples. In contrast, fumonisins (FBs), zearalenone (ZEN), and deoxynivalenol (DON) were detected in 100, 93, and 24% of TMR samples at mean levels of 336.7 ± 36.98, 80.32 ± 16.06 µg/kg and 292.1 ± 85.68 µg/kg, respectively. Ninety-two percent of TMR samples exhibited co-occurring mycotoxins. In milk, 54% of samples (n = 38) had detectable levels of mycotoxin, while 43% (n = 30) contained two or more types of mycotoxins. DON, FB, and ZEN metabolites (α-zearalenol and β-zearalenol) were the most frequent mycotoxins detected in the milk samples analyzed, at mean concentrations of 0.562 ± 0.112, 2.135 ± 0.296 µg/kg, 2.472 ± 0.436 µg/kg, and 0.343 ± 0.062 µg/kg, respectively. However, none of the analyzed milk samples had levels higher than the maximum permitted limit for AFM1 in Brazil (0.5 µg/L). The results of this trial highlight the concern about the co-occurrence of multiple mycotoxins in TMR and in milk, due to the possible additive or synergistic effects of these compounds. The presence of co-occurring mycotoxins in milk underscores the need for stringent preventive practices to avoid mycotoxin contamination in the diet of dairy cows in Brazil.

The High-Efficiency Degradation of Multiple Mycotoxins by Lac-W Laccase in the Presence of Mediators

Mycotoxin cocontamination is a severe threat to health and economic security worldwide. The mycotoxins aflatoxin B1 (AFB1), zearalenone (ZEN), deoxynivalenol, T-2 toxin, fumonisin B1, and ochratoxin A are of particular concern due to their substantial toxicity. Lac-W is a laccase with the unique property of degrading these six mycotoxins in the absence of redox mediators. Nevertheless, their degradation rates are low. This work aims to improve the ability of Lac-W to degrade these six mycotoxins and to elucidate its detoxification mechanism. Including redox mediators increased the Lac-W degradation efficiency drastically, and completely degraded AFB1 and ZEN within one hour. Additionally, Lac-W-AS has good temperature, pH, and ions adaptability in ZEN degradation. Lac-W-AS reduced the ZEN toxicity because ZEN degradation products significantly restored the bioluminescence intensity of Vibrio fischeri. A Lac-W-AS-mediated oxidation product of ZEN was structurally characterized as 15-OH-ZEN by UHPLC-MS/MS. Linear sweep voltammetry showed that AS affected the potential of Lac-W and accelerated the oxidation of ZEN. Finally, the combination of mediators (acetosyringone and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonate)) improved the degradation rate of mycotoxins. This work highlights that the combination of Lac-W with mediators serves as a good candidate for degrading multi-mycotoxin contaminants in food and feedstuff.

Toxicity of Low-Level Multiple-Mycotoxin Mixture in Nile Tilapia (Oreochromis niloticus) Is Prevented with Organically Modified Clinoptilolite Feed Additive

Organically modified clinoptilolite (member of the zeolite family of minerals; MinazelPlus®) feed additive, with an average weight of 30 ± 2 g, was used to prevent mycotoxicosis in Nile tilapia (Oreochromis niloticus) through its supplementation for 42 days to the diet contaminated with multiple mycotoxins: aflatoxin B1 (40 µg/kg), fumonisin B1 and B2 (600 µg/kg), zearalenone (50 µg/kg), and deoxynivalenol (150 µg/kg). The fish were divided randomly into four experimental groups (basal diet control—C; fed 2 g/kg MinazelPlus®—MZ; fed multiple mycotoxins—MT; and fed a combination of MinazelPlus® and multiple mycotoxins—MZ + MT). Each group consisted of triplicate aquarium setups, with six fish in each replicate. Sampling was performed in weeks 2, 4, and 6. The lymphocyte count was significantly higher in the MZ group compared with the MT group and the MT + MZ group in week 6. An overall decrease in the neutrophil count was observed in the experimental groups. Histopathological analysis was performed in weeks 2 and 6, revealing significant changes in the liver, intestines, kidney, and spleen of fish from the MT group, while the MT + MZ and MZ groups were similar to the control. The addition of 2 g/kg MinazelPlus® has the ability to prevent and reduce the adverse effects of chronic exposure to low concentrations of multiple mycotoxins in juvenile Nile tilapia.

Simultaneous detection of multiple mycotoxins in agricultural products: Recent advances in optical and electrochemical sensing methods.

Mycotoxin contamination poses serious threats to human and animal health. Food and environmental systems are often simultaneously contaminated with multiple mycotoxins, a problem that is further exacerbated by the synergistic toxicological effects of these co-occurring mycotoxins. Consequently, the development of rapid detection methods capable of simultaneously identifying multiple mycotoxins in agricultural products is essential to prevent their entry into the food chain. Compared to standard detection methods, optical and electrochemical (EC) sensing methods have distinct advantages for the rapid detection of mycotoxins. This review comprehensively summarizes the latest advancements in the field of simultaneous detection of multiple mycotoxins using optical and EC sensing methods over the last 6years (2018-2024). First, the review introduces the classification and relevant principles of optical and EC sensing methods. Thereafter, it emphasizes innovative simultaneous detection strategies within these approaches. Finally, it discusses current challenges and offers a reference for further research. Currently, the main challenge lies in the mutual interference among targets, making the development of an interference-free detection platform essential. Furthermore, the ongoing development of integrated technology is expected to aid regulatory authorities in improving the quality of agricultural products for field applications.

Rapid high-throughput screening of multiple typical mycotoxins in cereals

Rapid high-throughput screening of multiple typical mycotoxins in cereals

Individual and combined occurrences of the prevalent mycotoxins in commercial feline and canine food.

Mycotoxins, such as aflatoxin B1 (AFB1), 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 AFB1, 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, AFB1, and FBs, whereas ZEN was less frequently detected. The mean concentrations of the six mycotoxins in pet feed samples were 3.17μg/kg for AFB1, 0.65mg/kg for DON, 2.15mg/kg for FBs, 6.27μg/kg for OTA, 20.00μg/kg for T-2, and 30.00μg/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. AFB1, 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 AFB1, 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.

The Pathology and Blood Biochemistry of Juvenile Lates calcarifer on Diets Contaminated With Mycotoxins, Histamines and Rancid Fats-A Case Study.

Mycotoxins, originating from contaminated raw materials or suboptimal feed storage, are a growing concern in tropical aquaculture. Common fungi such as Aspergillus spp. and Fusarium spp. produce mycotoxins including aflatoxin, fumonisin, deoxynivalenol and zearalenone. High doses or prolonged exposure (weeks) to low doses of these mycotoxins (< 20 μg/kg) can depress growth, immunity, and cause mortality. This study investigated poor growth and low survival in juvenile Asian seabass, Lates calcarifer fed two different diets (FM40 and ABS3) for 5 weeks. Analysis of these diets revealed high peroxide values, multiple mycotoxins and high histamine levels. Fish fed the FM40 diet, which was contaminated with aflatoxin B1 (13.2 μg/kg), aflatoxin B2 (1.9 μg/kg), deoxynivalenol (29.5 μg/kg), alternariol (2.2 μg/kg), elevated peroxide value (45.91 mEq/kg), and histamine (129.51 mg/kg) developed mild bile duct hyperplasia, depressed total serum proteins (50.40 ± 10.06 g/L), markedly elevated blood potassium (8.2 ± 0.18 mmol/L), and heavy iron deposits in splenic melanomacrophage centres (Perl's stain) indicative of increased haemolysis. The presence of multiple cytotoxic mycotoxins in FM40 diet could explain the increased haemolysis and elevated blood potassium. In contrast, fish fed the ABS3 diet, which had high histamine levels (210.05 mg/kg), exhibited protein-losing nephropathy with multifocal fibrin plugs (Martius scarlet blue stain) indicating acute renal damage, and elevated blood calcium and phosphorus levels. Histamine is metabolised and excreted through the kidneys and known to induce renal arteriolar constriction, disrupt glomerular filtration barrier and increase permeability resulting in protein loss. This study shows that blood biochemistry and histopathology are useful diagnostic tools for assessing the impact of mycotoxins and histamines on fish health.

Photonic Microbead Array Digital Time-Resolved Fluorescence Ultrasensitive Platform for Simultaneous Detection of Multiple Mycotoxins.

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 AFB1), low limits of detection (LODs) (7.9 pg/mL for DON, 18 pg/mL for OTA, and 7.7 pg/mL for AFB1), 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.

Double detection of mycotoxins based on aptamer induced Fe3O4@TiO2@Ag Core − Shell nanoparticles “turn on” fluorescence resonance energy transfer

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 Fe3O4@TiO2 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 (R2 = 0.997) and 0.54 ng·mL−1 (R2 = 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.

Human health risk from co-occurring mycotoxins in dairy: A feed-to-fork approach

The presence of multiple mycotoxins in bovine dairy products can lead to potentially harmful health effects. This study examined the risk to human health from multiple mycotoxins in bovine dairy products using a feed-to-fork approach. It conducted a component-based risk assessment, using the toxicity of the most potent mycotoxin as a reference for others. The model also accounted for the influence of varying feed composition on mycotoxin contamination, considering monthly feeding pattern changes in Ireland. The probabilistic risk assessment uses Monte Carlo simulation to capture uncertainty and variability present in the model. The risk characterisation was undertaken using the Margin Of Exposure (MOE) method, where if the MOE was below 10,000, then the population's health was considered at risk. This study investigated the risk to human health from single mycotoxins and a combination of mycotoxins. The results showed that all mycotoxins were of low risk. However, the risk increased when aflatoxin was included in the co-occurring groups. Since 5% of the MOE was less than 10,000, the Irish population (male and female) was at relatively low risk of consuming aflatoxin or other co-occurring mycotoxin-contaminated milk. In comparison, the age group 8-12 years were at a relatively higher risk, with 10% of the MOE less than 10,000. The carry-over rates, the concentration of mycotoxin in feed components and the feeding composition were the most sensitive parameters. An increase in aflatoxin production in feed components can occur under climate scenarios and thus needs to be monitored, preventing any risks from aflatoxin or a combination of co-occurring mycotoxins. This study underscores the need for continuous monitoring and preventive measures, especially in feed components that are sensitive to aflatoxin-producing fungi that may contaminate bovine feed.

Simultaneous detection of multiple mycotoxins in Radix Dipsaci and estimation of exposure risk for consumers

Like many traditional Chinese herbal medicines, preparations from Radix Dipsaci are at risk of contamination by harmful mycotoxins; however, there have been no reports of actual contamination. In this study, we developed an analytical method to simultaneously detect eight mycotoxins in Radix Dipsaci and estimate the exposure risk for consumers. We have developed an analytical method utilizing ultra-high performance liquid chromatography and tandem mass spectrometry to accurately determine the levels of AFB1, AFB2, AFG1, AFG2, OTA, ZEN, T-2 and ST mycotoxins in 45 batches of Radix Dipsaci sourced from major medicinal herb markets across five regions in China. We also analyzed migration of mycotoxins from the raw herbs into water decoction. Based on these results and data on human consumption of the herbal medicine, we estimated risk of exposure and acceptable exposure limits to mycotoxins in the Radix Dipsaci using the “margin of exposure (MOE)” method. Of the 45 batches of Radix Dipsaci, 48.89% contained at least one of the eight mycotoxins, 24.44% contained one, 17.78% contained two and 6.67% contained three. The most frequent mycotoxins were aflatoxin B1, present in 35.56% of batches (at 0.25–34.84 μg/kg); aflatoxin G1, 15.56% (1.99–44.05 μg/kg); and ochratoxin A, 22.22% (16.11–143.38 μg/kg). These three mycotoxins transferred from the raw herb into water decoction at respective rates of 20.20%, 29.14%, and 24.80%. The 95th percentile values of the MOE risk factors for health effects of AFB1 were below 10,000 at high doses but above 10,000 at low doses of Radix Dipsaci long-term treatment. With the reduction in duration of exposure years, the MOE values of AFB1 and AFG1 gradually reverted to within the acceptable range. The mean, 50th, and 95th percentile values of the MOE risk factors for health effects of OTA exceeded 10,000 regardless of whether consumers received a low or high dose of Radix Dipsaci treatment for durations ranging from 1 to lifetime. Based on this exposure and a typical human diet, we have estimated the respective 20-year exposure limits for Radix Dipsaci to be 5.821 μg/kg, 4.035 μg/kg, and 56.073 μg/kg for the three mycotoxins under consideration. Contamination with multiple mycotoxins is frequently observed in Radix Dipsaci, and the three most prevalent contaminants have been found to leach into water decoctions, thereby posing a potential health hazard for individuals consuming this herbal preparation. This work highlights the need to monitor herbal medicines for mycotoxin contamination in order to protect consumers.

Establishment of appropriate conditions for the efficient determination of multiple mycotoxins in tea samples and assessment of their drinking risks

Conditions were determined for rapid, convenient, and efficient determination of 16 common mycotoxins in tea samples. Mycotoxins in tea leaves and tea infusion samples were extracted using solid-liquid extraction/liquid-liquid extraction combined with ultrasonic-assisted extraction. The extraction solvent was 2-butanone/ethyl acetate (9/1 v/v) with 0.1 % formic acid. The established conditions enabled the analysis of these mycotoxins by ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) in 5.5 min. In addition, HPLC with a temperature-controlled fluorescence detector was able to simultaneously determine 8 mycotoxins with fluorescent properties in 10 min without derivatization. Aflatoxin M1 (2.15 and 3.01 μg/kg), fumonisin B2 (198.89 μg/kg), and zearalenone (87.54 μg/kg) could be detected in commercially available pu-erh tea, green tea, and black tea products, and their total transfer rates from the products to brewed tea infusions were 64.08–65.13 %, 90.59 %, and 25.99 %, respectively. The risks of drinking mycotoxins from these tea infusions mostly showed low levels of concern.

First evidence on the occurrence of multi-mycotoxins and dietary risk exposure to AFB1 along the cassava value chain in Uganda

This study investigated the occurrence and distribution of multiple mycotoxins (aflatoxin B1, B2, G1, G2, fumonisins B1, B2, ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEN), and citrinin (CIT)) in cassava products and as assessed the potential risk of aflatoxin B1 (AFB1) exposure among cassava consumers. A total of 192 samples of cassava products (96 flour and 96 chips, each with 48 samples from farmer and 48 from wholesaler) were analysed using LC/MS–MS. All positive samples irrespective of their origin (flour or chips) exhibited AFB1 levels exceeding the EU regulatory threshold of 5 µg/kg. The sum of fumonisins (FB1 + FB2), ZEN, and DON were significantly (P < 0.05) higher in cassava flour (14.3 µg/kg; 3.71 µg/kg; 25.1 µg/kg) compared to chips (6.54 µg/kg; 1.25 µg/kg; 0.25 µg/kg), respectively. Aflatoxins G2 was not detected in any of 192 samples. Cassava flour samples from farmers exhibited significantly (P < 0.05) higher mean concentrations of AFB1 (27.1 µg/kg), total aflatoxins (78.2 µg/kg), and ochratoxin A (79.6 µg/kg) in contrast to wholesalers, whose mean levels were notably lower at 8.91, 5.79 µg/kg, and 2.44 µg/kg, respectively, pointing the likely critical source of mycotoxin contamination. Cassava consumers in Northern Uganda are at a higher risk, with an estimated 2.06 cancer cases per 100,000 individuals per year compared to those in Eastern Uganda at 0.25. This study underscores the urgent need for interventions to manage aflatoxins in cassava flour, particularly at farm level in Northern Uganda. It accentuates a shift market to household-level sampling and the need for analytical methods targeting multiple mycotoxins.

DNA template triggered copper nanoparticles generation coupled with hybridization chain reaction amplification for simultaneous detection of three mycotoxins in food samples

The co-occurrence of various mycotoxins in food, resulting from fungal infections, can lead to increased toxicity and enhanced harm through synergistic effects. In this work, a fluorescent assay was developed for simultaneous detection of zearalenone (ZEN), ochratoxin A (OTA) and aflatoxin B1 (AFB1) to address the issue of multiple mycotoxins co-contamination. Specific DNA hairpin-based aptamers were employed to recognize the mycotoxins and form a double-stranded DNA structure. Exonuclease III was introduced for the DNA cleavage, releasing single-stranded DNA triggers that could activate the assembling of DNA quadrilateral template. Copper nanoparticles were generated on the frame of DNA quadruplex template and produced fluorescent detection signals, while two hybridization chain reactions occurring on the different branches of the template and provided two additional detection signals. Inadequately reacted DNA chains were efficiently adsorbed and eliminated by Fe3O4@graphene oxide through magnetic separation. The limits of detection and linear ranges for ZEN, AFB1 and OTA were determined as 0.008 ng/mL, 0.003 ng/mL, 0.006 ng/mL and 0.010–100.000 ng/mL, 0.010–100.000 ng/mL, 0.010–50.000 ng/mL, respectively. Moreover, successful verification for real samples yielded recoveries from 99.2% to 102.7%, with a relative standard deviation of <5.23%. Artificially prepared toxic fungus infected samples were also analyzed. This method has demonstrated practical implications for the monitoring of food contaminants.

Mycotoxin contamination and the nutritional content of corn targeted for animal feed

Mycotoxin contaminated corn poses a risk to poultry production. Although mycotoxin regulatory guidelines are based on the hazards of individual mycotoxin contamination, feed and feed ingredients may be contaminated with multiple mycotoxins. The objective of this study was to assess mycotoxin co-contamination and its impact on the nutrient content of corn grain. Corn samples (n = 328) originating from various regions in the Southeastern U.S. were quantitatively analyzed for fumonisin (FUM), deoxynivalenol (DON), aflatoxin (AFB1) and zearalenone (ZEA) by HPLC-MS/MS. Nutritional content was analyzed by near-infrared spectroscopy, and color data were collected. All 328 samples were found to be contaminated with at least 1 mycotoxin: 100% contained FUM (19-24,680 µg/kg), 69.82% contained DON (0-9,640 µg/kg), 17.07% contained AFB1 (0-939 µg/kg), and 43.60% had detectable levels of ZEA (0-8,093.5 µg/kg). Most of the samples were contaminated with 2 or more mycotoxins, with only 18.29% of the samples containing a single mycotoxin. 38.41% of the samples had 2 mycotoxins present, 36.59 % had 3 mycotoxins, and 4.88% of the samples had all 4 tested mycotoxins present. Samples contaminated with AFB1 had significantly lower fat (P = 0.007) and lightness (P = 0.007); samples contaminated with DON had significantly higher starch (P < 0.001) and lower protein (P < 0.001). Samples contaminated with FUM had significantly higher protein (P = 0.008) and moisture (P = 0.019) and lower starch (P < 0.001). ZEA contaminated samples had significantly lower starch (P = 0.034). A correlation was observed between mycotoxin contamination and altered nutrient content in corn. This study provides further evidence that co-contamination of mycotoxins is the norm in corn, and that mycotoxin contamination correlates with impacts on the nutrient profile of feed corn.

Shifts in Fusarium Communities and Mycotoxins in Maize Residues, Soils, and Wheat Grains throughout the Wheat Cycle: Implications for Fusarium Head Blight Epidemiology.

Fusarium Head Blight (FHB), predominantly caused by Fusarium species, is a devastating cereal disease worldwide. While considerable research has focused on Fusarium communities in grains, less attention has been given to residues and soil, the primary inoculum sources. Knowledge of Fusarium spp. diversity, dynamics, and mycotoxin accumulation in these substrates is crucial for assessing their contribution to wheat head infection and the complex interactions among Fusarium communities throughout the wheat cycle. We monitored six minimum-tillage wheat fields, with maize as the preceding crop, over two years. Soils, maize residues, and wheat grains were sampled at four stages. Fusarium composition was analyzed using a culture-dependent method, species-specific qPCR, and EF1α region metabarcoding sequencing, enabling species-level resolution. The Fusarium communities were primarily influenced by substrate type, accounting for 35.8% of variance, followed by sampling location (8.1%) and sampling stage (3.2%). Among the 32 identified species, F. poae and F. graminearum dominated grains, with mean relative abundances of 47% and 29%, respectively. Conversely, residues were mainly contaminated by F. graminearum, with a low presence of F. poae, as confirmed by species-specific qPCR. Notably, during periods of high FHB pressure, such as in 2021, F. graminearum was the dominant species in grains. However, in the following year, F. poae outcompeted F. graminearum, resulting in reduced disease pressure, consistent with the lower pathogenicity of F. poae. Source Tracker analysis indicated that residues were a more significant source of Fusarium contamination on wheat in 2021 compared to 2022, suggesting that F. graminearum in 2021 primarily originated from residues, whereas F. poae's sources of infection need further investigation. Additionally, multiple mycotoxins were detected and quantified in maize residues during the wheat cycle, raising the question of their ecological role and impact on the soil microbiota.

SERS aptasensor for simultaneous detection of ochratoxin A and zearalenone utilizing a rigid enhanced substrate (ITO/AuNPs/GO) combined with Au@AgNPs

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.

Ultrasensitive electrochemical sensing for simultaneous rapid detection of zearalenone and ochratoxin A in feedstuffs and foodstuffs

Mycotoxins are causing persistent worries regarding food safety worldwide due to their ability to elicit a wide range of harmful effects in both animals and humans. Approaches that can identify a single mycotoxin are not sufficient for addressing the needs of quality control of agricultural products since they are often co-contaminated by multiple mycotoxins. This paper presents the development of an electrochemical sensor that is free of biomolecules and is capable of simultaneously detecting zearalenone (ZEN) and ochratoxin A (OTA) through the incorporation of nickel oxide (NiO) with carboxylated carbon nanotubes (MWCNT(–COOH)).This is the first time that NiO–MWCNT(–COOH) based electrochemical sensor has achieved the simultaneous potential-resolved selective detection of ZEN and OTA. Under optimized conditions, good linear responses were observed in the range of 0.01–10.24 μg mL−1 for ZEN and 0.04–10.24 μg mL−1 for OTA with low detection limit of 6 and 15 ng mL−1, respectively. The proposed electrochemical sensor demonstrated good selectivity, reusability, reproducibility, and shelf-life for the sensitive detection of both mycotoxins. Moreover, the sensor was made utilizing screen-printing technology to demonstrate its potential for commercialization and practical application. Our biomolecule-free electrochemical sensor was successful in evaluating these mycotoxins in actual samples of corn flour and wheat flour, proving its promising application in monitoring ZEN and OTA co-contamination in agricultural products.

An automated magnetic beads-based chemiluminescence immunoassay system for simultaneous quantification of multi-mycotoxins in agricultural products

The coexistence of multiple mycotoxins poses a significant risk to the safety of agricultural products. Hence, a novel and automated magnetic beads-based chemiluminescence immunoassay (MCLIA) system was developed for the simultaneous quantification of typical multi-mycotoxins, including zearalenone (ZEN), Ochratoxin A (OTA), deoxynivalenol (DON), and aflatoxin B1 (AFB1). It comprises a compact chemiluminescence-based analyzer and a specially designed mycotoxin assay kit. Firstly, antimycotoxin monoclonal antibody (mAb) was coupled with alkaline phosphatase (AP), and the antigen of mycotoxin was then with biotin. After detailed optimization of this indirect competition immunoassay, all necessary reagents were preloaded and sealed into a specialized plastic reagent strip to prepare the assay kit. Secondly, a compact chemiluminescence-based analyzer was developed by programmatically integrating magnetic, thermal, and optical mechanisms. The MCLIA system facilitates the detection of mycotoxins in up to 8 samples simultaneously or various mycotoxins in different samples within 20 min, according to actual needs. The detection limits (LOD) for ZEN, OTA, DON, and AFB1 were 2.55, 0.64, 15.47, and 0.78 μg/kg, respectively, with the 50 % inhibition concentrations (IC50) in the ranges of 1.84−4.25, 0.95−1.69, 11.36−13.98, and 0.37−0.54 ng/mL. Parallel analysis of commercial agricultural products demonstrated a strong linear correlation (R2 = 0.9843–0.9962) with UPLC-HRMS, confirming the MCLIA system's reliability for the simultaneous quantitative detection of various mycotoxins. Offering advantages in detection sensitivity, dynamic range, and turnaround time, the compact MCLIA system facilitates automated and parallelized detection in the model of “sample-in to result-out”. It presents a versatile platform for the efficient immunodetection of other contaminants.

Oxidative degradation and detoxification of multiple mycotoxins using a dye-decolorizing peroxidase from the white-rot fungus Bjerkandera adusta

Mycotoxins are widely present from cereal crops to food products, posing a significant risk to food safety and causing serious health and economic losses. Therefore, it is imperative to develop a sustainable and efficient detoxification strategy to address mycotoxin contamination. Herein, a dye-decolorizing peroxidase, BaDyP, from the white-rot fungus Bjerkandera adusta was successfully expressed as a soluble form in Escherichia coli with the help of molecular chaperones. Purified recombinant BaDyP could efficiently degrade multiple mycotoxins including aflatoxin B1, zearalenone, and deoxynivalenol in the presence of mediators such as Mn2+ or 1-HBT. Meanwhile, their degradation products were identified using UPLC-MS/MS, including AFB1-diol, AFQ1, 15-OH-ZEN, HZEN, and C15H18O8. Furthermore, cell survival tests exhibited a significant reduction in the biological toxicity of these degradation products. These findings indicate that the use of dye-decolorizing peroxidase could be a promising approach for the effective removal of multiple mycotoxins in cereal crops and food products.