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Food contaminated by mycotoxins has become a worldwide public problem with political and economic implications. Although a variety of traditional methods have been used to eliminate mycotoxins from agri-foods, the results have been somewhat less than satisfactory. As an emerging non-thermal processing technology, atmospheric cold plasma (ACP) has great potential for food decontamination. Herein, this review mainly presents the degradation efficiency of ACP on mycotoxins in vitro and agri-foods as well as its possible degradation mechanisms. Meanwhile, ACP effects on food quality, factors affecting the degradation efficiency and the toxicity of degradation products are also discussed. According to the literatures, ACP could efficiently degrade many mycotoxins (e.g., aflatoxin, deoxynivalenol, zearalenone, ochratoxin A, fumonisin, and T-2 toxin) both in vitro and various foods (e.g., hazelnut, peanut, maize, rice, wheat, barley, oat flour, and date palm fruit) with little effects on the nutritional and sensory properties of food. The degradation efficacy was dependent on many factors including ACP treatment parameter, working gas, mycotoxin property, and food substrate. The mycotoxin degradation by ACP was mainly attributed to the reactive oxygen and nitrogen species in ACP, which can damage the chemical bonds of mycotoxins, consequently reducing the toxicity of mycotoxins., 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 Inc. All rights reserved.)

Fusarium species infect maize crops leading to the production of fumonisin by their toxigenic members. Elimination of microbes is critical in mitigating further postharvest spoilage and toxin accumulation. The current study investigates the efficacy of a previously described multispectral sorting technique to analyze the reduction of fumonisin and toxigenic Fusarium species found contaminating maize kernels in Kenya. Maize samples (n = 99) were collected from six mycotoxin hotspot counties in Kenya (Embu, Meru, Tharaka Nithi, Machakos, Makueni, and Kitui County) and analyzed for aflatoxin and fumonisin using commercial ELISA kits. Aflatoxin levels in majority (91%) of the samples were below the 10 ng/g threshold set by the Kenya Bureau of Standards and therefore not studied further. The 23/99 samples that had >2,000 ng/g of fumonisin were selected for sorting. The sorter was calibrated using kernels sourced from Ghana to reject visibly high-risk kernels for fumonisin contamination using reflectance at nine distinct wavelengths (470-1,550 nm). Accepted and rejected streams were tested for fumonisin using ELISA, and the presence of toxigenic Fusarium using qPCR. After sorting, there was a significant (p < 0.001) reduction of fumonisin, by an average of 1.8 log ng/g (98%) and ranging between 0.14 and 2.7 log ng/g reduction (28-99.8%) with a median mass rejection rate of 1.9% (ranged 0% to 48%). The fumonisin rejection rate ranged between 0 and 99.8% with a median of 77%. There was also a significant reduction (p = 0.005) in the proportion of DNA represented by toxigenic Fusarium, from a mean of 30-1.4%. This study demonstrates the use of multispectral sorting as a potential postharvest intervention tool for the reduction of Fusarium species and preformed fumonisin. The spectral sorting approach of this study suggests that classification algorithms based on high-risk visual features associated with mycotoxin can be applied across different sources of maize to reduce fumonisin., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Asha B. Mohamed reports financial support was provided by International Atomic Energy Agency. Editorial Board member of the Journal of Food Protection – MJS., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Nowadays, the companion animals (dogs or other pets) are considered as members of the family and have established strong emotional relationships with their owners. Dogs are long lived compared to food animals, so safety, adequacy, and efficacy of dog food is of great importance for their health. Cereals, cereal by-products as well as feedstuffs of plant origin are commonly employed food resources in dry food, yet are potential ingredients for mycotoxins contamination, so dogs are theoretically more vulnerable to exposure when consumed daily. Aflatoxins (AF), deoxynivalenol (DON), fumonisins (FUM), ochratoxin A (OTA), and zearalenone (ZEA) are the most frequent mycotoxins that might present in dog food and cause toxicity on the growth and metabolism of dogs. An understanding of toxicological effects and detoxification methods (physical, chemical, or biological approaches) of mycotoxins will help to improve commercial ped food quality, reduce harm and minimize exposure to dogs. Herein, we outline a description of mycotoxins detected in dog food, toxicity and clinical findings in dogs, as well as methods applied in mycotoxins detoxification. This review aims to provide a reference for future studies involved in the evaluation of the risk, preventative strategies, and clear criteria of mycotoxins for minimizing exposure, reducing harm, and preventing mycotoxicosis in dog., 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 The Authors. Published by Elsevier Inc. All rights reserved.)

Deoxynivalenol (DON) and fumonisin B 1 (FB 1 ), as a group of highly toxic secondary metabolites, have become a potential source of water environmental pollutants. To minimize two mycotoxins exposure to consumers, a dual lateral flow immunoassay (LFIA) integrated with the smartphone was reported for simultaneous and quantitative detection of DON and FB 1 in the water environment. The significantly improved sensitivity was contributed to a smartphone-based device with the ability to image and analyze results. Under optimized conditions, the detection limits of DON and FB 1 were calculated to be 3.46 and 2.65 ng/mL, which were approximately 25 and 10 folds lower than those of the visual detection of the LFIA. This method showed good specificity and a good dynamic linear detection for DON and FB 1 . The recoveries of DON and FB 1 were evaluated by the spiked lake water, river water, and pond water, ranging from 92.47% to 106.2% with the relative standard deviation under 9.13%. Moreover, the results of the developed LFIA showed a high correlation with enzyme-linked immunosorbent assay (ELISA) results, with a correlation coefficient of 0.999 for DON and 0.996 for FB 1 , respectively. To sum up, the developed LFIA provides a promising platform for sensitive, simultaneous, quantitative, and on-site detection of DON and FB 1 in the water environment., 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 © 2022. Published by Elsevier B.V.)

Fumonisins have posed hazardous threat to human and animal health worldwide. Enzymatic degradation is a desirable detoxification approach but is severely hindered by serious shortage of detoxification enzymes. After mining enzymes by bioinformatics analysis, a novel carboxylesterase FumDSB from Sphingomonadales bacterium was expressed in Escherichia coli, and confirmed to catalyze fumonisin B1 to produce hydrolyzed fumonisin B1 by liquid chromatography mass spectrometry for the first time. FumDSB showed high sequence novelty, sharing only ~34% sequence identity with three reported fumonisin detoxification carboxylesterases. Besides, FumDSB displayed its high degrading activity at 30-40 °C within a broad pH range from 6.0 to 9.0, which is perfectly suitable to be used in animal physiological condition. It also exhibited excellent pH stability and moderate thermostability. This study provides a FB1 detoxification carboxylesterase which could be further used as a potential food and feed additive., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020. Published by Elsevier B.V.)

This study describes a smart analysis platform capable of quantitative measurements using a multiplex lateral flow strip. Using the multi-mycotoxin strip, five fungal toxins were simultaneously and quantitatively detected in naturally contaminated wheat. First, a matrix-based standard curve was established for the detection of aflatoxin B1 (AFB1), fumonisin B1 (FB1), T-2, deoxynivalenol (DON), and zearalenone (ZEN). Established on an open android system, the platform is able to read 6 lines on the strip simultaneously. The platform is equipped with a Quick Response code scanning model, which reads the established standard curves, and then rapidly quantify mycotoxins in naturally contaminated wheat. All the data and sample information are stored on a central server through the platform which is linked to the cloud. The limits of detection (LOD) for AFB1, FB1, T-2, DON, and ZEN in wheat were 4, 20, 10, 200, and 40 μg/kg and the visual cut off values was 20, 1000, 200, 4000, and 400 μg/kg, separately. To validate the platform and the multi-mycotoxin detection method, 10 wheat samples were analyzed and the results were in a good agreement with those obtained by LC-MS/MS. The platform will be a powerful tool for crop monitoring services., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Mycotoxins, such as fumonisin B 1 (FB 1 ) and deoxynivalenol (DON), are toxic secondary metabolites produced by fungi. Herein, the simultaneous detection of FB 1 and DON (both of which are water-soluble) in grain samples by immunochromatographic methods was used to probe whether the levels of these mycotoxins exceeded the limit of 1 mg/kg. Two types of immunochromatographic test strips (ICSs) were developed for this purpose, namely, those based on Au nanospheres (prepared via reduction with trisodium citrate) and Au nanoflowers (prepared by Au seed-mediated growth) as markers, with the respective sensitivities to both FB 1 and DON equalling 20 and 5.0 ng/mL. The former ICS was used to detect FB 1 and DON in grain (51 wheat samples and 18 maize samples), providing results consistent with those of high-performance liquid chromatography and enzyme-linked immunosorbent assays. The developed technique was suitable for the on-site screening of large-scale samples for FB 1 and DON., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Abstract: A total of 133 samples of whole wheat and barley grains and wheat and barley flour collected from retail markets in the main cities of Punjab, Pakistan, were analyzed for the mycotoxin fumonisin B1 (FB1) using reverse phase high-performance liquid chromatography with fluorescence detection. Of these samples, 120 (90%) were positive for FB1, and 75 (63%) of the 120 positive samples had FB1 concentrations higher than the European Union maximum (200 μg/kg). The limit of detection was 4 μg/kg. The highest mean (±SD) concentration of FB1 was found in whole wheat samples, 980.5 ± 211.4 μg/kg. The calculated dietary intakes of FB1 from wheat and barley flours were 4,456 and 503.7 ng/g of body weight per day, respectively., (Copyright ©, International Association for Food Protection.)

The electrochemical interaction of mycotoxin, Fumonisin B1 (FB 1 ) with fish sperm double stranded DNA (fsDNA) was investigated by using electrochemical impedance spectroscopy (EIS) technique. The pencil graphite electrodes were used for impedimetric detection of fsDNA and FB 1 , and also the impedimetric monitoring of surface confined interaction of FB 1 with fsDNA under the optimized conditions. The experimental conditions; the concentration of DNA and FB 1 and the interaction time of FB 1 with fsDNA were firstly optimized. The main features of impedimetric DNA biosensor, such as its detection limit with the reproducubility were also discussed. The selectivity of detection of interaction between FB 1 and fsDNA was also explored in the mixture samples containing FB 1 and each of other mycotoxins, such as zearalenone (ZEN), deoxyvinalenol (DON) in order to examine the selectivity of impedimetric DNA biosensor., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Due to the widely occurring co-contamination of mycotoxins in raw food materials, simultaneous monitoring of multiple mycotoxins is needed. Herein, we report the design and fabrication of an electrochemical immunosensor for simultaneous detection of two mycotoxins, fumonisin B1 (FB1) and deoxynivalenol (DON), in a single test. A dual-channel three-electrode electrochemical sensor pattern was etched on a transparent indium tin oxide (ITO)-coated glass via photolithography and was integrated with capillary-driven polydimethylsiloxane (PDMS) microfluidic channel. The two working electrodes were functionalized with gold nanoparticles and anti-FB1 and anti-DON antibodies. Tests were performed by incubating the working electrodes in a sample solution introduced in the PDMS channel. The formation of toxin-antibody immunocomplexes on the working electrode surface produced electrochemical signal responses, which were recorded and compared with control signal to quantify individual mycotoxin concentrations. Using this dual-channel ITO-microfluidic electrochemical immunosensor we achieved limits of detection (LODs) of 97 pg/mL and 35 pg/mL, respectively for FB1 and DON, and their corresponding linear ranges of detection were 0.3-140 ppb and 0.2-60 ppb. The sensor performance, which remained stable for two weeks under proper storage, was validated by testing with ground corn extract used as a real food matrix., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A multiplex Lateral Flow Immunoassay was developed based on the use of a single Test line and multicolour gold nanoparticles (GNPs) as signal reporters. Red and blue GNPs were linked to antibodies directed towards two different analytes and included in a typical lateral flow immunoassay configuration, in which the Test line was formed by the mixture of two antigens. As a result of the immunoreactions occurring at the Test zone, diverse combinations of red and blue GNPs labels were captured. Therefore, the Test line assumed different colours depending on which - and how much - analyte is present in the sample. The multiplexing capability of the 'colour-encoded assay' is illustrated by the simultaneous detection of aflatoxin B1 (AFB1) and type-B fumonisins (FMs) in wheat and food products that made with wheat. Reproducible detection of AFB1 and FMs contamination in raw and processed food was achieved with visual cut-off levels at 1 ng mL -1 and 50 ng mL -1 , respectively. The contaminant was identified based on the colour of the label according with a specific colour code. Furthermore, strips images were acquired by means of a common smartphone and analysed through RGB data analysis providing semi-quantitative detection of the two mycotoxins., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A label-free optical biosensor for the fast simultaneous determination of three mycotoxins, aflatoxin B 1 (AFB 1 ), fumonisin B 1 (FB 1 ) and deoxynivalenol (DON), in beer samples is presented. The biosensor is based on an array of ten Mach-Zehnder interferometers (MZIs) monolithically integrated along with their respective broad-band silicon light sources onto a single chip. Multi-analyte determination is accomplished by functionalizing the sensing arms of individual MZIs with mycotoxin-protein conjugates. Assay is performed by pumping over the chip mixtures of calibrators or samples with a mixture of specific monoclonal antibodies, followed by reaction with a secondary anti-mouse IgG antibody. Reactions are monitored in real-time by continuously recording the MZI output spectra, which are then subjected to Discrete Fourier Transform to convert spectrum shifts to phase shifts. The detection limits achieved for AFB 1 , FB 1 and DON were 0.8, 5.6 and 24 ng/ml, respectively, while the assay duration was 12 min. Recovery values ranging from 85 to 115% were determined in beer samples spiked with known concentrations of the three mycotoxins. In addition, beers of different types and origin were analysed with the biosensor developed and the results were compared with those provided by established laboratory methods, further supporting the accuracy of the proposed device., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Naked-eye readout colorimetric signal-based assays are promising for high-throughput screening detection and point-of-care diagnostics in resource-constrained regions. Here, a novel direct competitive plasmonic enzyme-linked immunosorbent assay (dc-pELISA) based on gold nanoparticle (AuNP) aggregation with highly sensitive and robust naked-eye readout signal was developed and used to detect fumonisin B 1 (FB 1 ). AuNP aggregation was induced by a horseradish peroxidase (HRP)/hydrogen peroxide (H 2 O 2 )/tyramine (TYR) system, resulting in a dramatic color change from red to blue. In this system, H 2 O 2 was produced via GOx-mediated glucose oxidation reaction, and FB 1 -GOx was used as a competitive antigen. The proposed pELISA demonstrated good linear detection of FB 1 from 3.125 ng mL -1 to 25 ng mL -1 with a vivid color change from deep blue to red and a cutoff limit of 12.5 ng mL -1 observed by the naked eye. The average recoveries for FB 1 -spiked corn samples ranged from 76.5% to 96.8% with a relative standard derivation of 4.88~ 16.4%. Meanwhile, the proposed method exhibited excellent agreement (R 2 = 0.927) with the conventional ELISA method in blindly detecting FB 1 spiked corn samples. Additionally, the results of the proposed method for FB 1 positive corn samples also showed a high consistency with those of the ultra performance liquid chromatography method. These results indicated acceptable accuracy and precision of the proposed colorimetric ELISA for quantitative detection of FB 1 in actual corn samples., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Maize ( Zea mays) is a staple in many developing countries but is known to be prone to pest (insects, birds, and rodents) and fungal infestation. In Guatemala, mycotoxin contamination of cultivated products may occur owing to such factors as environmental conditions and the use of traditional agriculture operations. To assess the current maize conditions in Guatemala, a small-scale study was performed. Mold and insect counts and mycotoxin (aflatoxin and fumonisin) concentrations were determined on 25 farms in two townships (Chiantla and Todos Santos) of the Huehuetenango Department. Total fungal counts were 3.6 to 6.83 log CFU/g with no significant differences ( P > 0.05) across farms at different altitudes. Farms where maize was not produced but was purchased were at higher risk of fumonisin contamination, whereas local producers were mostly affected by aflatoxins. Aflatoxin was present in maize from 100% of farms at 1.0 to 85.3 ppb, and fumonisin was detected on 52% of farms at 0.4 to 31.0 ppm. Average mycotoxin consumption amounts were above the recommended maximum intake for aflatoxin in both produced and purchased maize and above the provisional maximum tolerable daily intake for fumonisin in purchased maize. Estimated daily intake was 0.01 to 0.85 μg/kg of body weight per day for aflatoxin and 2.9 to 310.0 μg/kg of body weight per day for fumonisin. An entomological analysis revealed overall 32% prevalence of Ephestia kuehniella (flour moth), 16% prevalence of Sitophilus zeamais (maize weevil), and 8% prevalence of Tribolium sp. (flour beetle) on the analyzed farms. This study highlighted poor agricultural practices used in the highlands of Guatemala. Current practices should be revised for the production of maize that is safe for consumption by the population in this region.

Mycotoxins are secondary fungi metabolites present in foods that cause adverse effects in humans and animals. The aims of this work were to develop and validate a multi-mycotoxin method to determine the presence of aflatoxins, citreoviridin, deoxynivalenol, fumonisins ochratoxin A, zearalenone and some metabolites/derivatives in rice, maize-based products and wheat-based products, and a method to determine total fumonisins (free and bound/hidden forms) in maize-based products. The validated multi-mycotoxin method was based on extraction with acidified acetonitrile and LC-ESI + -MS/MS analysis, with LOQs ranging from 0.5 to 121μg/kg, and proved to be suitable for the multi-mycotoxin analysis in wheat, maize and rice products. Bound/hidden fumonisins were determined after extraction of the free forms using the multi-mycotoxin method, followed by a basic hydrolysis of the unextracted bound/hidden and solid-liquid extraction with low temperature purification (SLE-LTP). The hydrolysis efficiency of the bound/hidden extraction procedure was investigated by analyzing a maize reference material and showed recoveries ranging from 75% (HFB2) to 108% (HFB1). The use of isotope internal standards was crucial for mycotoxins quantification in maize meal and wheat flour, while for rice, external calibration and matrix matched curves gave satisfactory results. All 55 samples of wheat-based products analyzed were contaminated with at least one mycotoxin and 16% of 44 rice samples were also contaminated. The most prevalent mycotoxins were DON and ZON in wheat-based products., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Fusarium mycotoxins such as trichothecenes, zearalenone and fumonisins occur on a worldwide basis in cereal grains, animal feeds and forages. Practical solutions for multiple mycotoxin determination in samples are required by industry and regulators for cost effective screening purposes. The feasibility of developing a novel multiplex nanoarray for the simultaneous and semi-quantitative detection of three regulated mycotoxins: zearalenone (ZEA), T2-toxin (T2) and fumonisin B1 (FUM) was examined. Additionally, the assay was also able to detect HT2 toxin and fumonisin B2 and B3 due to the cross reactivity profiles of the antibodies used. Individual mycotoxin conjugates specific to the three mycotoxins were nano-spotted onto wells of a microtitre plate. Optimisation of assay parameters and antibodies was undertaken with both individual and multiplex calibration curves generated. A competitive assay format was employed enabling a calibration curve for concentration analysis and duplicate results for up to 40 samples in 70min for the three target mycotoxins. The characteristics and performance of the nanoarray were evaluated including sensitivity and specificity for each target. Additionally, intra and inter spotting precision, cross reactivity, matrix effects and sample analysis in maize and wheat (n=8) was performed. Sensitivity, determined as the concentration causing 50% inhibition, was 70.1, 2.8 and 90.9ppb in PBS, 172.4, 3.2 and 129.3ppb in methanol, 197.4, 0.7 and 216.7ppb in wheat and 43.6, 0.5 and 25.9ppb in maize for ZEA, T2 and FUM respectively. Intra spotting precision was 6%, 11% and 10% for PBS and 5%, 11% and 12% for methanol for ZEA, T2 and FUM respectively. Inter spotting precision was 4%, 14% and 6% for PBS and 3%, 9% and 16% for methanol for ZEA, T2 and FUM respectively. The feasibility of the nanoarray as an easy to use sensitive screening tool in the 96 well format has been demonstrated for the multiplex detection of three regulated mycotoxins. Improvements in automated image and data analysis software for novice end users are required to improve the overall rapidity of analysis., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In this paper, a novel highly sensitive chemiluminescence immune-affinity 96 spots monolith array was developed to detect deoxynivalenol (DON), zearalenone (ZEN), T-2 toxin (T-2), and fumonisin B1 (FB1) in corn samples. Firstly, the monolith array was prepared through on suit UV-initiated copolymerization using polyethylene glycol diacrylate (PEGDA) as cross-linker, glycidyl methacrylate (GMA) as functional monomer and polyethylene glycol 200 (PEG 200) as the porogen. Subsequently, the four mycotoxins immune-affinity monolith array was prepared by immobilization of DON, ZEN, T-2, and FB1 antibody. The mole ratio of PEGDA/GMA, UV exposure time, and the volume ratio of PEG 200/PEGDA were optimized to improve the performances of the immune-affinity monolith array. For the mycotoxins immune-affinity monolith array based on chemiluminescence detection, the limit of detection was 0.0036ng/mL (DON), 0.0048ng/mL (ZEN), 0.0039ng/mL (T-2), and 0.0017ng/mL (FB1), respectively. The linear response in the range of 0.01-0.1ng/mL (R(2)=0.98). The results showed that the proposed four mycotoxins immune-affinity monolith array was a stable, accurate, and highly sensitive method to determine levels of DON, ZEN, T-2, and FB1 in real samples., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Fusarium verticillioides is a major maize pathogen and there are susceptible and resistant cultivars to this fungal infection. Recent studies suggest that its main mycotoxin fumonisin B1 (FB1) may be involved in phytopathogenicity, but the underlying mechanisms are mostly still unknown. This work was aimed at assessing whether FB1 disseminates inside the plants, as well as identifying possible correlations between the maize resistant/susceptible phenotype and the unbalances of the FB1-structurally-related sphingoid base sphinganine (Sa) and phytosphingosine (Pso) due to toxin accumulation. Resistant (RH) and susceptible hybrid (SH) maize seedlings grown from seeds inoculated with a FB1-producer F. verticillioides and from uninoculated ones irrigated with FB1 (20 ppm), were harvested at 7, 14 and 21 days after planting (dap), and the FB1, Sa and Pso levels were quantified in roots and aerial parts. The toxin was detected in roots and aerial parts for inoculated and FB1-irrigated plants of both hybrids. However, FB1 levels were overall higher in SH seedlings regardless of the treatment (infection or watering). Sa levels increased substantially in RH lines, peaking at 54-fold in infected roots at 14 dap. In contrast, the main change observed in SH seedlings was an increase of Pso in infected roots at 7 dap. Here, it was found that FB1 disseminates inside seedlings in the absence of FB1-producer fungal infections, perhaps indicating this might condition the fungus-plant interaction before the first contact. Furthermore, the results strongly suggest the existence of at least two ceramide synthase isoforms in maize with different substrate specificities, whose differential expression after FB1 exposure could be closely related to the susceptibility/resistance to F. verticillioides., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

A fluorescent immunochromatographic strip test (ICST) based on the use of Quantum Dots (QD) was developed and applied to detect fumonisins in maize samples. A limit of detection for fumonisin B1 of 2.8 µg L(-1) was achieved, with an analytical working range of 3-350 µg L(-1), corresponding to 30-3500 µg kg(-1) in maize flour samples, according with the extraction procedure. The time required to perform the analysis was 22 min, including sample preparation. Recovery values in the range from 91.4% to 105.4% with coefficients of variation not exceeding 5% were obtained for fortified and naturally contaminated maize flour samples. To evaluate the possible improvements due to the use of QD for ICST technology, we performed a direct comparison of the proposed QD-ICST to a gold nanoparticles- and a chemiluminescent-ICST previously developed for fumonisins detection, in which the same immunoreagents were employed., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Nanobodies that are small and thermally stable, as well as have high expression level, are leading alternative to produce anti-idiotypic antibodies. These antibodies have the advantage of replacing mycotoxins and their conjugates for immunoassays. In this work, anti-fumonisin B1 (FB1) monoclonal antibody (mAb) was used as the target for biopanning from a naïve alpaca nanobody (Nb) phage display library. After three cycles of panning, one anti-idiotypic nanobody (Ab2β Nb) was isolated and subjected to a Nb-ELISA for the detection of FB1. Surface plasmon resonance was used to analyze the reaction kinetics between the Ab2β Nb and anti-FB1 mAb. The developed assay showed a half inhibitory concentration (IC50) of 0.95±0.12 ng/mL, a limit of detection of 0.15 ng/mL, a linear range of 0.27-5.92 ng/mL, and a low cross-reactivity toward FB2 of 4.93%. The sensitivity was enhanced approximately 20-fold compared with that of the chemosynthetic FB1-BSA conjugates. The equilibrium dissociation constant (KD) measured for Ab2β Nb: anti-FB1 mAb was 164.6 nM. The assay was compared with conventional ELISA (the commercial ELISA kit), and the results indicated the reliability of Ab2β Nb replacing the antigen-carrier protein conjugates. The use of biotechnology in developing the surrogate is an ideal strategy for replacing conventional synthesized antigens., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The B-series fumonisins (FBs) are some of the most prevalent mycotoxins produced as a secondary metabolite by Fusarium species growing on cereals. For decades they have been studied extensively in food and feed products, but there is no information about their occurrence in the aquatic environment or about how these mycotoxins are transported to the surface water and the groundwater. The aim of this study was to clarify the causes of fumonisin occurrence in aquatic ecosystems by examining the relation between mycotoxin contamination of crops and their levels in the aquatic environment. Water samples were collected from drainage ditches and wells or watercourses located in agricultural areas in the Wielkopolska region, Poland. Our research conducted on an annual basis showed the seasonal variability of fumonisin B1 concentration in the analyzed water samples, with the highest concentration in the post-harvest season (September to October) at 48.2 ng L(-1), and the lowest in winter and spring at 21.9 ng L(-1). Fumonisins B2 and B3 in water samples were not detected. Cereal samples were collected in the harvest season from each agricultural area close to tested water bodies. Mycotoxins were present in all cereal samples at concentrations from 43.3 to 1055.9 ng g(-1). Our results confirm that fumonisins are transported to aquatic systems by rainwater through soil. On the basis of available literature, this is the first report concerning the presence of fumonisin B1 in different aquatic environments. To date their ecotoxicological effects are largely unknown and require further investigation., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This paper describes the use of two immunoaffinity columns (IACs) coupled in tandem, providing selective clean-up, based on targeted mycotoxins known to co-occur in specific matrices. An IAC for aflatoxins+ochratoxin A+fumonisins (AOF) was combined with an IAC for deoxynivalenol+zearalenone+T-2/HT-2 toxins (DZT); an IAC for ochratoxin A (O) was combined with a DZT column; and an aflatoxin+ochratoxin (AO) column was combined with a DZT column. By combining pairs of columns it was demonstrated that specific clean-up can be achieved as required for different matrices. Samples of rye flour, maize, breakfast cereal and wholemeal bread were analysed for mycotoxins regulated in the EU, by spiking at levels close to EU limits for adult and infant foods. After IAC clean-up extracts were analysed by LC-MS/MS with quantification using multiple reaction monitoring. Recoveries were found to be in range from 60 to 108%, RSDs below 10% depending on the matrix and mycotoxin combination and LOQs ranged from 0.1n g/g for aflatoxin B1 to 13.0 ng/g for deoxynivalenol. Surplus cereal proficiency test materials (FAPAS(®)) were also analysed with found levels of mycotoxins falling within the satisfactory range of concentrations (Z score ≤ ± 2), demonstrating the accuracy of the proposed multi-mycotoxin IAC methods., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Fumonisin B analogs, particularly FB1, FB2, and FB3, are major mycotoxins found in cereals. Single-chain fragment variable (scFv) antibodies represent a promising alternative immunoassay system. A phage-displayed antibody library derived from four monoclonal antibodies (mAbs) generated against FB1 was used to screen high binding affinity scFv antibodies; the best candidate was designated H2. Surface plasmon resonance measurements confirmed that the H2 scFv displayed a 82-fold higher binding affinity than its parent mAb. Direct competitive enzyme-linked immunosorbent assay demonstrated that the H2 antibody could competitively bind to free FB1, FB2, and FB3, with an IC50 of 0.11, 0.04, and 0.10 μM, respectively; it had no cross-reactivity to deoxynivalenol, nivalenol and aflatoxin. Validation assays with naturally contaminated samples revealed a linear relationship between the H2 antibody-based assay results and chemical analysis results, that could be expressed as y=1.7072x+5.5606 (R(2)=0.8883). Homology modeling of H2 revealed a favorable binding structure highly complementary to the three fumonisins. Molecular docking analyses suggested that the preferential binding of the H2 scFv to FB2 was due to the presence of a hydrogen radical in its R1 position, leading to a proper electrostatic matching and hydrophobic interaction. The H2 scFv antibody can be used for the rapid, accurate, and specific detection of fumonisin contamination in agricultural samples., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The genus Aspergillus section Nigri, or the black aspergilli, represents genetically closely related species that produce the mycotoxins, ochratoxins and the fumonisins. Fumonisin B1 (FB1) is of an added concern because it is also a virulence factor for maize. Our preliminary data indicated that black aspergilli could develop asymptomatic infections with maize and peanuts plants. Symptomless infections are potential problems, because under favorable conditions, there is a potential for accumulation of ochratoxins and the fumonisins in contaminated postharvest crops. In the present report, the ability of black aspergilli from peanuts and maize to produce ochratoxin A and FB1 on maize kernels was assessed. One hundred fifty strains from peanuts and maize were isolated from several southeastern and midwestern states. Aspergillus nigri (A. nigri var. nigri) was the dominant species (87%), while Aspergillus foetidus, Aspergillus japonicus, Aspergillus tubingensis, and Aspergillus carbonarius were infrequently isolated. None of the wild isolates produced detectable amounts of ochratoxins. However, we do report the occurrence of the fumonisins B1, B2, and B3. Of 54 field isolates, 30% (n = 16) produced FB1, 61% (n = 33) produced FB2, and 44% (n = 24) produced FB3. The amounts of fumonisins produced during the test period of 30 days suggest that these strains might be weak to moderate producers of fumonisin on maize. To our knowledge, this is a first report of FB1 and FB3 production by isolates of black aspergilli from an American cereal and legume.

Unsanitary conditions during harvesting, drying, packing and storage stages in production and processing of spices and herbs could introduce mycotoxin contamination. The occurrence of ochratoxin A and fumonisins in popular spices and herbs was studied, using liquid chromatography-electrospray-mass spectrometry. Apart from mycotoxins, ergosterol as a factor indicating fungal development was also analysed. A total of 79 different samples commercialized in Poland were randomly purchased from popular markets were tested for mycotoxins. The frequency of samples with fumonisins was lower (31%) than ochratoxin A (49%). Free from mycotoxins were samples of bay leaf and white mustard. ERG content - in spice samples with high concentration level of mycotoxins - was also significantly higher than in samples with little to no mycotoxins., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Powdery mildew (PM), caused by the fungus Erysiphe necator, is one of the most widespread fungal disease of grape and may cause extensive openings on the berry surface during the infection. We evaluated the effect of damage caused by PM in grape berries on the growth of and mycotoxin production by Aspergillus and on the oxidative stress in infected berries. Berries of Vitis vinifera L. cv. Negroamaro with sound skin (SS) and those naturally infected by PM were surface sterilized and inoculated with either fumonisin B2(FB2)-producing strains of Aspergillus niger or ochratoxin A (OTA)-producing strains of Aspergillus carbonarius and incubated at 20 and 30°C. The PM berries were significantly more susceptible to both Aspergillus colonization (5 to 15 times more susceptible) and OTA and FB2 contamination (2 to 9 times more susceptible) than were SS berries. The highest toxin concentration was detected in inoculated PM berries both for OTA (9 ng/g) at 20°C and for FB2 (687 ng/g) at 30°C. In inoculated SS and PM berries, although malondialdehyde and hydrogen peroxide concentrations did not increase, the two black Aspergillus species caused a significant decrease in ascorbate content, thus inducing a pro-oxidant effect. These results indicate that grape berries affected by PM are more susceptible to black Aspergillus growth and to production and/or accumulation of FB2 and OTA. Thus, preventive control of E. necator on grape berries could reduce the mycotoxicological risk from black Aspergillus infection.

In this work, we presented a new aptasensor for fumonisin B1 (FB1) based on fluorescence resonance energy transfer (FRET) between NaYF4: Yb, Ho upconversion fluorescent nanoparticles (UCNPs) and gold nanoparticles (AuNPs). The quenchers (AuNPs) were attached to the 5' end of the molecular beacon (MB), and the donors (UCNPs) were attached to the 3' end of the MB. In the absence of target DNA (DNA complementary to FB1 aptamers), the energy donors and acceptors were placed in close proximity, leading to quenching of the fluorescence of the UCNPs. Due to the combination of FB1 and FB1-specific aptamers, this caused some complementary DNA dissociating from the magnetic nanoparticles (MNPs). In the presence of the complementary DNA, the MBs underwent spontaneous conformational change and caused the UCNPs and AuNPs to detach from each other, resulting in restoration of the upconversion fluorescence. Therefore, the fluorescence of UCNPs was restored in a FB1 concentration-dependent manner, which was the basis of the FB1 quantification. The aptasensors showed a linear relationship from 0.01 to 100 ng mL(-1) for FB1 with a detection limit of 0.01 ng mL(-1) in an aqueous buffer. As a practical application, the aptasensor was used to monitor FB1 levels in naturally contaminated maize samples. The results were consistent with that of a classic ELISA method, indicating that the UCNPs-FRET aptasensor, which benefited from the near infrared excitation of NaYF4: Yb, Ho UCNPs, was effective for directly sensing FB1 in foodstuff samples without optical interference. This work also created the opportunity to develop aptasensors for other targets using this FRET system., (Copyright © 2013 Elsevier B.V. All rights reserved.)

In a previous study, inedible almond pick-out samples were assayed for aflatoxin and aflatoxigenic Aspergillus species. These samples contained high populations of black-spored Aspergillus section Nigri species. To investigate whether these species may contribute to the total potential mycotoxin content of almonds, Aspergillus section Nigri strains were isolated from these samples and assayed for ochratoxin A (OTA) and fumonisin B2 (FB2). The majority of isolates (117 strains, 68%) were identified as Aspergillus tubingensis, which do not produce either mycotoxin. Of the 47 Aspergillus niger and Aspergillus awamori isolates, 34 strains (72%) produced FB2 on CY20S agar, and representative strains produced lower but measurable amounts of FB2 on almond meal agar. No OTA-producing strains of Aspergillus section Nigri were detected. Almond pick-out samples contained no measurable FB2, suggesting that properly dried and stored almonds are not conducive for FB2 production by resident A. niger and A. awamori populations. However, 3 of 21 samples contained low levels (<1.5 ng/g) of OTA, indicating that sporadic OTA contamination may occur but may be caused by OTA-producing strains of other Aspergillus species.

Background: Exposure to multiple mycotoxins through the food chain represents a major potential health hazard to both humans and livestock. They can cause a variety of severe acute as well as chronic diseases. Eliminating mycotoxins from various grain crops is a global health priority. According to the Food and Agriculture Organization (FAO), world food production needs to double by 2050. Innovative solutions will be required to sustain toxin free grain supplies worldwide., Methods: A competitive flow cytometry based multiplexed assay with fluorescent microspheres has been developed. The new multiplexed method can analyze simultaneously any one or all six major mycotoxins. They include: Ochratoxin A (OTA), Aflatoxin B1 (AFB1), Fumonisin B1 (FB1), T-2 toxin (T-2), Deoxynivalenol (DON) and Zearalenone (ZEA), which are all potential human health hazards. The CFIA described here includes a simplified single extraction step for mycotoxins from specimens and a comprehensive post acquisition software module. The new assay system was developed with a FACSArray™ BD Bioanalyzer flow cytometer (BD Biosciences, Belgium)., Results: The CFIA performs favourably when compared to commercial ELISA. Sensitivity range with CFIA increased between 13% and 100% with an average improvement of 50% for the six mycotoxins., Conclusions: The multiplexed assay presented here has the unique capacity to quantify up to six mycotoxins simultaneously from a single specimen extraction. CFIA's poly-mycotoxin detection sensitivity exceeds standard ELISA. CFIA may be part of a comprehensive assay system that will provide reliable and effective safeguard for agricultural commodities to be free of mycotoxins., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Fusarium verticillioides (teleomorph Gibberella moniliformis) is the main fungal agent of ear and kernel rot of maize (Zea mays L.) worldwide, including Italy. F.verticillioides is a highly toxigenic species since it is able to produce the carcinogenic mycotoxins fumonisins. In this study, 25 F. verticillioides strains, isolated from maize in different regions of Italy were analyzed for their ability to produce fumonisins, their pathogenicity and their genetic variability. A further referenced strain of G. moniliformis isolated from maize in USA was also used as outgroup. The fumonisins B₁, B₂, and B₃ were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Pathogenicity tests were carried out by symptom observation and determination of growth parameters after inoculation of maize seeds, seedlings and wounded detached leaves. Total genomic DNA was used for Amplified Fragment Length Polymorphism (AFLP) analysis. About 20% of the analyzed strains were unable to produce fumonisins in in vitro experiments on inoculated maize flour, while, among fumonisin producers, a great variability was observed, with values ranging from 1 to 115 mg kg⁻¹. The different analyzed strains showed a wide range of pathogenicity in terms of effect on seed germination, seedling development and of symptoms produced on detached leaves, which were not correlated with the different in vitro fumonisin production. AFLP analysis indicated the presence of genetic diversity not only between the Italian strains and the American reference but also among the Italian isolates., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Fumonisins are mycotoxins produced by various species of Fusarium and occur naturally in contaminated maize and maize-based foods. Ingestion of fumonisins has considerable health implications for humans and animals. Since fumonisins lack a useful chromophore or fluorophore, their determination in maize is routinely achieved via HPLC with fluorescence detection (FLD) after precolumn derivatization. This study optimized naphthalene-2,3-dicarboxaldehyde (NDA) derivatization of fumonisins in naturally contaminated maize following strong anion exchange (SAX) solid phase extraction (SPE) clean-up and utilizing diode array detection (DAD) as a practical alternative simultaneously to FLD. The limit of detection (LOD) for fumonisin B(1) (FB(1)), fumonisin B(2) (FB(2)) and fumonisin B(3) (FB(3)) with FLD was 0.11 ng, 0.50 ng and 0.27 ng, respectively, and with DAD it was 13.8 ng, 12.5 ng and 6.6 ng, respectively injected on column. The coefficient of variation (CV, n = 6) for FB(1), FB(2) and FB(3) in a naturally contaminated samples obtained with FLD was 2.6%, 1.8% and 5.3%, respectively, compared to 6.0%, 3.4% and 9.5%, respectively, obtained with DAD. Subsequently the optimized NDA derivatization was compared to the widely used o-phthaldialdehyde (OPA) derivatization agent as well as alternative sample clean-up with immunoaffinity column (IAC) by analyzing naturally contaminated maize samples (n = 15) ranging in total fumonisin (TFB = FB(1)+FB(2)+FB(3)) levels from 106 to 6000 μg/kg. After immunoaffinity column clean-up of extracted samples, the recoveries of spiked maize samples for NDA-FLD of FB(1), FB(2) and FB(3) were 62%, 94% and 64%, respectively. NDA proved to be an effective derivatization reagent of fumonisin in naturally contaminated maize samples following IAC clean-up, except for DAD at TFB levels below 1000 μg/kg. In contrast NDA derivatization following SAX clean-up produced results comparable to OPA only for levels below 1000 μg/kg. Aside from the difference in detection limits, FLD and DAD produced comparable results irrespective of the clean-up method or the derivatization agent., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A total of 255 corn samples collected in 2010 from three main corn production provinces of China (Liaoning, Shandong, and Henan) were analyzed for contamination with fumonisins (FB(1) and FB(2)). The incidence of contamination was significantly higher in samples from Liaoning than in samples from the other two provinces. Approximately 80.0% of the samples from Liaoning were contaminated with fumonisins, with a mean total fumonisin concentration of 3,990 ng/g. In contrast, the mean total fumonisin concentrations were 845 and 665 ng/g in samples from Shandong and Henan, respectively. The probable daily intake of fumonisins (0.3 μg/kg of body weight) is within the provisional maximum tolerable daily intake of 2.0 μg/kg of body weight set by the Joint Food and Agriculture Organization and World Health Organization Expert Committee on Food Additives.

Fungi belonging to Aspergillus section Nigri occur frequently and in high populations on grapes. Species within this section include Aspergillus niger, A. tubingensis, and A. carbonarius, and they are potential sources for mycotoxins including ochratoxin A and fumonisin B(2) (FB(2)) in grapes and grape products. Aspergillus section Nigri strains were isolated from California raisins to examine the frequency and extent of FB(2) production. Of 392 strains isolated, 197 strains were identified as A. niger, 131 of which produced FB(2). These strains produced from 1.2 to 27 μg/ml FB(2) in culture. PCR amplification of fum1 and fum19 gene fragments showed that all FB(2)-producing strains and nearly all nonproducing strains of A. niger contain these genes. An additional 175 strains were identified as A. tubingensis, none of which produced FB(2). PCR with fum1 and fum19 primers amplified gene fragments of 14 and 25% of A. tubingensis strains, respectively, suggesting that putative orthologs of A. niger fumonisin biosynthetic genes might occur in A. tubingensis. These results indicate that FB(2) production is common among field isolates of A. niger and suggest that the potential for FB(2) contamination of California raisins should be addressed further.

A quantitative lateral flow immunoassay for measuring fumonisins in maize was developed. Strip preparation and assay parameters were optimized to obtain a dipstick usable outside the laboratory with different samples, and which shows performance comparable with that of other screening methods, as confirmed by the intra- and the inter-day precision of data (RSD 5-16%). Quantification was obtained by an external calibration curve, which can be stored and used for measurements made with strips of the same batch in different days and at varying temperatures (22-37°C). Limit of detection (120 μgL(-1)) and dynamic range (200-5000 μgL(-1)) allow the direct assessment of fumonisin contamination at all levels of regulatory relevance. Twenty-seven maize samples were analyzed after a simple sample preparation which avoids the use of organic solvent. Linear correlation was observed (y=1.071x-0.2, r(2)=0.990) when data was compared with that obtained through a reference LC-MS/MS method, across a wide range of fumonisin contamination., (Copyright © 2010 Elsevier B.V. All rights reserved.)

The development of a reversed phase high performance liquid chromatography fluorescence method for the determination of the mycotoxins fumonisin B(1) and fumonisin B(2) by using silica-based monolithic column is described. The samples were first extracted using acetonitrile:water (50:50, v/v) and purified by using a C(18) solid phase extraction-based clean-up column. Then, pre-column derivatization for the analyte using ortho-phthaldialdehyde in the presence of 2-mercaptoethanol was carried out. The developed method involved optimization of mobile phase composition using methanol and phosphate buffer, injection volume, temperature and flow rate. The liquid chromatographic separation was performed using a reversed phase Chromolith(®) RP-18e column (100 mm × 4.6 mm) at 30 °C and eluted with a mobile phase of a mixture of methanol and phosphate buffer pH 3.35 (78:22, v/v) at a flow rate of 1.0 mL min(-1). The fumonisins separation was achieved in about 4 min, compared to approximately 20 min by using a C(18) particle-packed column. The fluorescence excitation and emission were at 335 nm and 440 nm, respectively. The limits of detections were 0.01-0.04 μg g(-1) fumonisin B(1) and fumonisin B(2), respectively. Good recoveries were found for spiked samples (0.1, 0.5, 1.5 μg g(-1) fumonisins B(1) and B(2)), ranging from 84.0 to 106.0% for fumonisin B(1) and from 81.0 to 103.0% for fumonisin B(2). Fifty-three samples were analyzed including 39 food and feeds and 14 inoculated corn and rice. Results show that 12.8% of the food and feed samples were contaminated with fumonisin B(1) (range, 0.01-0.51 μg g(-1)) and fumonisin B(2) (0.05 μg g(-1)). The total fumonisins in these samples however, do not exceed the legal limits established by the European Union of 0.8 μg g(-1). Of the 14 inoculated samples, 57.1% contained fumonisin B(1) (0.16-41.0 μg g(-1)) and fumonisin B(2) (range, 0.22-50.0 μg g(-1)). Positive confirmation of selected samples was carried out using liquid chromatography-tandem mass spectrometry, using triple quadrupole analyzer and operated in the multiple reaction monitoring mode., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Between 2004 and 2007 we examined foods from Japanese retail shops for contamination with ochratoxin A (OTA) and fumonisins B(1), B(2), and B(3). A total of 1,358 samples of 27 different products were examined for OTA, and 831 samples of 16 different products were examined for fumonisins. The limits of quantification ranged from 0.01 to 0.5 microg/kg for OTA and 2 to 10 microg/kg for the fumonisins. OTA was detected in amounts higher than limits of quantification in wheat flour, pasta, oatmeal, rye, buckwheat flour and dried buckwheat noodles, raisins, wine, beer, coffee beans and coffee products, chocolate, cocoa, and coriander. OTA was found in more than 90% of the samples of instant coffee and cocoa, and the highest concentration of OTA, 12.5 microg/kg, was detected in raisins. The concentration of OTA in oatmeal, rye, raisins, wine, and roasted coffee beans varied remarkably from year to year. Fumonisins were detected in frozen and canned corn, popcorn grain, corn grits, cornflakes, corn soups, corn snacks, beer, soybeans, millet, and asparagus. The highest concentrations of fumonisins B(1), B(2), and B(3) were detected in corn grits (1,670, 597, and 281 microg/kg, respectively). All of the samples of corn grits were contaminated with fumonisins, and more than 80% of the samples of popcorn grain and corn snacks contained fumonisins. OTA and fumonisins were detected in several food products in Japan; however, although Japan has not set regulatory levels for these mycotoxins, their concentrations were relatively low.

Fumonisins are mycotoxins mainly produced by Fusarium verticillioides, which is a major contaminant of corn. However, there are sporadic reports of fumonisin contamination in wheat worldwide. The rice adherent fungus Gibberella fujikuroi is taxonomically closely related to F. verticillioides. Therefore, the potential risk of fumonisin contamination in rice and wheat is significant. Previously, a sensitive detection method utilizing liquid chromatography with tandem electrospray mass spectrometry (LC-ESI-MS-MS) was developed for the determination of fumonisins in brown rice. In the present study, the incidence of fumonisins in brown rice and wheat harvested in Japan was investigated using LC-ESI-MS-MS. Forty-eight rice samples and 47 wheat samples were screened and analyzed for the major B-type fumonisins: fumonisin B1 (FB1) and fumonisin B2 (FB2). About 1 kg of rice or wheat seed was divided into three subsamples, and 10 g from each subsample was used for the analysis. The limits of detection were 0.012 and 0.011 mg/kg for FBt and FB2, respectively, in rice samples and 0.010 and 0.008 mg/kg for FB1 and FB2, respectively, in wheat samples. The mean (standard deviation) recoveries of FB1 spiked at 0.50 mg/kg into toxin-free rice and wheat samples were 77.6 (4.2)% and 84.5 (3.1)%, respectively. One of the wheat samples was positive for FBt with a value greater than the limit of detection,but no fumonisin was found in any of the rice samples. This is the first report of fumonisins detected in Japanese wheat.

To survey fumonisins B1 (FB1) and B2 (FB2) in agricultural products consumed in South Korea and provide an exposure assessment, ground samples were extracted (80% MeOH), filtered (0.2 microm), and cleaned up. After evaporation, dry residues were reconstituted in 50% MeOH, and a 50-micro1 aliquot of this sample was mixed with 200 micro1 of o-phthaldialdehyde for derivatization. The derivatives were analyzed with a high-performance liquid chromatography system equipped with a fluorescence detector. For validation of the detection procedure, linearity, accuracy, precision, detection limit, and quantification limit were determined. The validated detection method was then used to survey fumonisins in white rice, brown rice, barley, barley tea, beer, wheat flour, millet, dried corn, corn flour, corn tea, canned corn, popcorn, and breakfast cereal. Retention times for FB1 and FB2 standards were 7 and 18 min, respectively. Linearity (R2 = 0.99995 to 0.99998), accuracy (81.47 to 108.83%), precision (2.35 to 5.77), detection limit (25 ng/g or ng/ml), and quantification limit (37 ng/g or ng/ml) indicated that this procedure is capable of quantifying fumonisins in agricultural products. Only FB1-positive samples (5.12%, three dried corn samples and five corn flour samples) were found at 90.89 to 439.67 ng/g. According the survey results, an estimated daily intake of FB1 and FB2 in Korea was 0.087 ng/kg of body weight per day. These results indicate that continuous monitoring of these mycotoxins is necessary to establish appropriate risk assessment, and the maximum tolerable daily intake of fumonisins in Korea is lower than the 2 microg/kg set by the Joint Food and Agriculture Organization-World Health Organization Expert Committee.

A sensitive and reliable method using pressurized liquid extraction (PLE) and liquid chromatography (LC)/electrospray ionization (ESI) tandem mass spectrometry with a triple quadrupole (QqQ) analyzer has been developed for the analysis of fumonisin B(1) (FB(1)), fumonisin B(2) (FB(2)) and fumonisin B(3) (FB(3)) in corn-based baby foods. Influence of several extraction parameters that affect PLE efficiency such as temperature, pressure, solvent extraction, number of cycles and dispersant/clean-up agents were studied. The selected PLE operating method was: 3g of sample was packed into 11 ml stainless-steel cell and fumonisins were extracted with methanol at 40 degrees C, 34 atm in one cycle of 5 min at 60% flush. The analytes were ionized in ESI operating with positive ion mode and identified by selecting two monitoring transitions, permitting quantification and confirmation in a single injection. Recoveries ranged from 68% to 83% at fortification levels of 200 microg kg(-1) with relative standard deviation (RSD) from 4% to 12%. The limits of quantification were from 2 microg kg(-1) for FB(1) and FB(2), and 5 microg kg(-1) for FB(3), which are below the maximum residue level established by the European Union legislation in infant formulas. The proposed method was successfully applied to the analysis of twenty seven samples of baby food products collected from different markets, and one positive sample with a content of 15.9 microg kg(-1) for FB(1), 9.2 microg kg(-1)for FB(2) and 5.8 microg kg(-1) for FB(3) was obtained. Given the simplicity and potential of the proposed procedure, its application for safety control is recommended.

A sensitive and selective analytical method was developed for the quantitative determination of fumonisins B(1) and B(2) in maize-based foods for direct human consumption. The method, based on high-performance liquid chromatography and fluorescence detection, presents a rapid and automated on-line post-column derivatization, performed with o-phtalaldehyde and N,N-dimethyl-2-mercaptoethylamine. Several factors affecting the separation and detection of fumonisins were investigated, including mobile phase composition, column features, derivatization agent flow-rate and both the excitation and the emission wavelengths. Optimal fluorescence detection was obtained by using a lambda(exc) of 343 nm and a lambda(em) of 445 nm. Under the optimized experimental conditions, a complete separation of fumonisins was obtained in less than 13 min by using a C(18) column and a gradient elution at 0.8 mL/min with methanol and 0.1M phosphate buffer at pH 3.15. The limits of detection for FB(1) and FB(2) were 4 and 5 microg/L corresponding to 5 and 6 microg/kg in matrix. Each fumonisin was determined in the range 40-320 microg/L that correspond to 50-400 microg/kg in matrix. The necessary requirements for accuracy, reproducibility and sensitivity were fulfilled and recovery values ranged from 87 to 94% for FB(1) and from 70 to 75% for FB(2) in cornflake samples at three fortification levels in the range 100-300 microg/kg. The potential of this method, combined with a simple clean-up procedure, was assessed by the measurements of FB(1) and FB(2) in maize-based products, such as maize flour, "polenta", tortillas and cookies.

Dried figs from Turkey that were visibly moldy (or fluorescent under UV light) and thus rejected as unsuitable for human food were screened for the presence of fungal metabolites. Crude solvent extracts from individual figs were directly analyzed by liquid chromatography combined with time-of-flight mass spectrometry to generate accurate mass data for all detectable components. A comparison of these data with a metabolite database indicated the presence of fumonisins B2 and B4, patulin, HT-2 toxin, and zearalenone among various other metabolites. Portions of the same figs were reextracted and then analyzed by conventional liquid chromatography-mass spectrometry. On the basis of coincident retention times and by matching selected ion monitoring for coincident ions with that of authentic standards, the identification of fumonisin B2, HT-2 toxin, patulin, and zearalenone was confirmed.

Sixty samples of corn from both conventional and organic farms were tested for internal fungal contamination. Molds were identified to genus, and those belonging to the genus Fusarium were identified to species. Twenty isolates of Fusarium verticillioides were tested with a high-performance liquid chromatography-naphthalene dicarboxaldehyde-fluorescence method for their ability to produce fumonisins B1 and B2. The internal fungal infection in organic maize (63.20%) was significantly higher than that in conventional maize (40.27%) (P < 0.05). However, the distribution of fungal genera indicated a significantly higher prevalence of Fusarium in conventional (34.93%) than in organic (18.15%) maize, making Fusarium the predominant fungus in conventional maize. This difference in mold distribution between organic and conventional maize was attributed to the difference in cultivation system. The dominant Fusarium species in both conventional and organic samples was F. verticillioides. There were no significant differences in the ability of 20 selected isolates of F. verticillioides to produce fumonisins on conventional or organic corn. Up to 13.3% of the conventional corn samples contained fumonisins B1 and B2 at mean concentrations of 43 and 22 ng/g, respectively. Organic corn samples had somewhat lower levels of contamination: 35 ng/g fumonisin B1 and 19 ng/g fumonisin B2 (P > 0.05). The organic farming system, with well-balanced crop rotation, tillage, and compost fertilization, produced corn that was less likely to be contaminated with Fusarium species, although no significant difference in fumonisin concentrations was found between the two types of contaminated corn.

Mycotoxins are toxic secondary metabolites produced by filamentous fungi that are growing on agricultural commodities. Their frequent presence in food and their severe toxic, carcinogenic and estrogenic properties have been recognised as potential threat to human health. A reliable risk assessment of mycotoxin contamination for humans and animals relies basically on their unambiguous identification and accurate quantification in food and feedstuff. While most screening methods for mycotoxins are based on immunoassays, unambiguous analyte confirmation can be easily achieved with mass spectrometric methods, like gas chromatography/mass spectrometry (GC/MS) or liquid chromatography/mass spectrometry (LC/MS). Due to the introduction of atmospheric pressure ionisation (API) techniques in the late 80s, LC/MS has become a routine technique also in food analysis, overcoming the traditional drawbacks of GC/MS regarding volatility and thermal stability. During the last few years, this technical and instrumental progress had also an increasing impact on the expanding field of mycotoxin analysis. The aim of the present review is to give an overview on the application of LC-(API)MS in the analysis of frequently occurring and highly toxic mycotoxins, such as trichothecenes, ochratoxins, zearalenone, fumonisins, aflatoxins, enniatins, moniliformin and several other mycotoxins. This includes also the investigation of some of their metabolites and degradation products. Suitable sample pre-treatment procedures, their applicability for high sample through-put and their influence on matrix effects will be discussed. The review covers literature published until July 2006.

We conducted a survey of aflatoxin B1, B2, G1, and G2, ochratoxin A, and fumonisin B1, B2, and B3 contamination in various foods on the retail market in Japan in 2004 and 2005. The mycotoxins were analyzed by high-performance liquid chromatography, liquid chromatography-mass spectrometry, or high-performance thin-layer chromatography. Aflatoxins were detected in 10 of 21 peanut butter samples; the highest concentration of aflatoxin B1 was 2.59 microg/kg. Aflatoxin contamination was not found in corn products, corn, peanuts, buckwheat flour, dried buckwheat noodles, rice, or sesame oil. Ochratoxin A was detected in oatmeal, wheat flour, rye, buckwheat flour, green coffee beans, roasted coffee beans, raisins, beer, and wine but not in rice or corn products. Ochratoxin A concentrations in contaminated samples were below 0.8 microg/kg. Fumonisins were detected in popcorn, frozen corn, corn flakes, and corn grits. The highest concentrations of fumonisins B1, B2, and B3 in these samples were 354.0, 94.0, and 64.0 microg/kg, respectively.

Fumonisin B1 (FB1) and fumonisin B2 (FB2) are the main members of a family of mycotoxins produced by various fungal species belonging to the Gibberella fujikuroi complex. The present work shows the results of a comparative study of various clean-up and derivatization procedures for analysis of fumonisins in rice cultures. Fumonisins were extracted from rice with acetonitrile/water (50/50, v/v). For clean-up, three solid-phase extraction procedures were assayed (C18 cartridge, SAX cartridge, and a combination of both). Two reagents (o-phthaldialdehyde and 4-fluoro-7-nitro-benzofurazan) were studied comparatively for formation of fluorescent derivatives. The separation was carried out by LC using a fluorescence detector. The best procedure for analysis of fumonisins in rice involved clean-up with C18 cartridge and derivatization with o-phthaldialdehyde. The limit of detection was 0.010 mg kg(-1) for both toxins. In the 10-500 mg kg(-1) spiking level range, the recovery rates for FB1 and FB2 in rice varied from 94.6% to 103.6% and from 96.3% to 101.9%, respectively. The optimized analytical method for determination of fumonisins in rice was applied to the study of FB1 and FB2 production by four isolates of the G. fujikuroi species complex in rice cultures carried out at different temperatures and water activities to establish the influence of strain and environmental conditions on fumonisin production in this cereal. In general, fumonisin production was the highest at 20 degrees C and lowest at 37 degrees C. Four of the five assayed water activity (aw) values (0.97, 0.98, 0.99, and 1.0) did not affect significantly fumonisin accumulation but fumonisins were not detected in cultures when aw was 0.96.

The effect of mobile phase pH, column temperature, extraction method and derivatisation time on the fluorescence response and recovery of fumonisin B1 in corn was investigated. Column temperature and mobile phase pH were negatively and positively correlated with the fluorescence response, respectively. Use of an Ultraturrax blender for extraction resulted in higher FB1 recoveries compared to a rotary shaker. In contrast to other reports, maximum fluorescence response occurred after a derivatisation time of 8 min. These results reflect on the absolute requirement for standardisation of the aforementioned parameters.

The effects of prolonged oral administration of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) mycotoxins were evaluated in broiler chickens from 21 to 42 d of age. A total of 192 birds were housed in experimental batteries and assigned to 32 cages, 6 birds per cage. The following treatments were applied: 1) 0 mycotoxins (control), 2) 10 mg of FB1, 3) 50 microg of AFB1, 4) 50 microg of AFB1 + 10 mg of FB1, 5) 350 microg of AFB1, 6) 350 microg of AFB1 + 10 mg of FB1, 7) 2,450 microg of AFB1, 8) 2,450 microg of AFB1 + 10 mg of FB1/kg of feed. Each treatment consisted of 4 replicates of 6 birds each. At the end of the trial, blood samples from 12 birds per treatment were collected, and the birds were necropsied. Compared with controls, the percentage of heterophils was lower (P < 0.05) in birds from groups receiving 50 microg of AFB1/kg + 10 mg of FB1/ kg and 2450 microg of AFB1/kg alone or in combination with FB1. A higher percentage of lymphocytes (P < 0.05) was observed in birds fed 50 microg of AFB1/kg + 10 mg of FB1/ kg, 350 microg of AFB1/kg, and 2,450 microg of AFB1/kg. A decrease in plasma albumin was observed only in birds fed 2,450 microg of AFB1/kg + 10 mg of FB1/kg. The liver of AFB1-treated birds had focal areas of necrosis and inflammatory infiltrates. In birds fed rations containing only 10 mg of FB1/kg, bile duct hyperplasia with fibrosis and a mononuclear infiltrate accompanied by trabecular derangement were observed. In contrast, in treatments in which FB1 was administered in combination, hepatic vacuolar degeneration was observed, and renal tissue presented corpuscles with increased cellular agglomeration, characterizing glomerulonephritis, and a clearly visible tubular epithelium with areas of degeneration and necrosis. The FB1 residues were detected in liver and in excreta of all FB1-treated groups, at levels that ranged from 0.013 to 0.051 mg/kg and from 1.19 to 2.79 mg/kg, respectively. Results indicated that FB1 and AFB1, singly or in combination at the levels evaluated, do not change markedly the hematological and serological parameters of broiler chickens, but may cause relevant lesions in liver and in kidneys.

Fusarium species are fungi that infect maize products worldwide and elaborate mycotoxins, which have been associated with cancer. This study was carried out to investigate the potential of lactic acid bacteria fermentation in reducing mycotoxin concentration and toxicity in maize meal products. Maize meal was spiked separately with fumonisin B1 and zearalenone and then allowed to ferment for 4 days. The potential cytotoxicity of the mycotoxin-spiked fermented extracts was also investigated using the SNO human esophageal carcinoma cell line (the SNO cell line was explanted from a cancer patient, S.N., a 62-year-old Zulu man, in July 1972). A significant decrease (P < 0.05) in the concentration of the two mycotoxins was observed, with a 56 to 67% and a 68 to 75% reduction in the third and fourth days, respectively. The two mycotoxins were not detectable in commercially fermented maize meal (amahewu) samples. After fermentation, mycotoxin-spiked maize meal samples containing lactic acid bacteria culture were comparatively less toxic to SNO cells than were samples without lactic acid bacteria. However, this difference in toxicity was not significant (P > 0.05). These results indicate that lactic acid bacteria fermentation can significantly reduce the concentration of mycotoxins in maize. However, such a reduction may not significantly alter the possible toxic effects of such toxins. The exact mechanism of toxin reduction warrants further investigation.