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Starches-rich and protein-rich cereal samples commonly need tedious sample preparation steps before instrumental analysis. This study developed a miniaturized centrifugal integrated cold-induced phase separation (CIPS) method for convenient sample preparation. A small-sized centrifuge tube (2 mL) and a low-temperature centrifuge, both of which are easily accessible, make up the basic components of the system. Unlike conventional sample preparation methods that need a step-by-step extraction, enrichment, purification, and centrifugation, this centrifugal integrated CIPS method can be performed by a one-step combination protocol under a low-temperature centrifuge. As a proof-of-concept study, satisfactory recoveries and enrichment factors were demonstrated for the extraction of fumonisins and ochratoxins from cereals. A sensitive and selective quantification method was yielded by combining LC-HRMS using tSIM acquisition mode, with good linearity (R 2  > 0.998), accuracy (82.9-106.5%), and precision (<13.4%). This strategy is convenient, low-cost, repeatable, and easy to semi-automate, further expanding the extraction potential for other acidic 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 Elsevier Ltd. All rights reserved.)

Among cereal contaminants, mycotoxins are of concern due to their importance in terms of food and feed safety. The difficulty in establishing a diagnosis for mycotoxicosis relies on the fact that the effects are most often subclinical for chronic exposure and the most common scenario is multi-contamination by various toxins. Mycotoxin co-occurrence is a major food safety concern as additive or even synergic toxic impacts may occur, but also regarding current regulations as they mainly concern individual mycotoxin levels in specific foods and feed in the food chain. However, due to the large number of possible mycotoxin combinations, there is still limited knowledge on co-exposure toxicity data, which depends on several parameters. In this context, this systematic review aims to provide an overview of the toxic effects of two regulated mycotoxins, namely ochratoxin A and fumonisin B1. This review focused on the 2012-2022 period and analysed the occurrence in Europe of the selected mycotoxins in different food matrices (cereals and cereal-derived products), and their toxic impact, alone or in combination, on in vitro intestinal and hepatic human cells. To better understand and evaluate the associated risks, further research is needed using new approach methodologies (NAM), such as in vitro 3D models. KEY CONTRIBUTION: Cereals and their derived products are the most important food source for humans and feed for animals worldwide. This manuscript is a state of the art review of the literature over the last ten years on ochratoxin A and fumonisin B1 mycotoxins in these products in Europe as well as their toxicological effects, alone and in combination, on human cells. Future perspectives and some challenges regarding the assessment of toxicological effects of mycotoxins are also discussed., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Fumonisins comprise structurally related metabolites mainly produced by Fusarium verticillioides and Fusarium proliferatum . Contamination with fumonisins causes incalculable damage to the economy and poses a great risk to animal and human health. Fumonisins and their covert products are found in cereals and cereal products. Food processing significantly affects the degradation of toxins and the formation of covert toxins. However, studies on fumonisins and their covert mycotoxins remain inadequate. This review aims to summarize changes in fumonisins and the generation of covert fumonisins during processing. It also investigates the toxicity and determination methods of fumonisins and covert fumonisins, and elucidates the factors affecting fumonisins and their covert forms during processing. In addition to the metabolic production by plants and fungi, covert fumonisins are mainly produced by covalent or noncovalent binding, complexation, or physical entrapment of fumonisins with other substances. The toxicity of covert fumonisins is similar to that of free fumonisins and is a non-negligible hazard. Covert fumonisins are commonly found in food matrices, and methods to analyze them have yet to be improved. Food processing significantly affects the conversion of fumonisins to their covert toxins.

Objective: To monitor fumonisins(FBs) in grains and grain products in Zhejiang and assess the exposure risks of FBs to local residents., Methods: Liquid chromatography coupled with tandem mass spectrometry method was used to determine the occurrence of FBs in rice, millet, dried noodles, instant noodles, and maize grains, and food frequency questionnaires were used to collect the food consumption data of Zhejiang population. Then, the simple probability distribution model was used to assess the exposure risk., Results: The levels of FBs in rice, millet, dried noodles and instant noodles were relatively low. The occurrence of FB&#95;1, FB&#95;2 and FB&#95;3 in these foods was 0-23.7%, 0-16.7% and 0-5.4%, respectively, and the mean levels were not detected(ND)-22.36, ND-20.63 and ND-7.19 μg/kg correspondingly. However, the levels of FBs in maize grains were relatively high. The occurrence of FB&#95;1, FB&#95;2, and FB&#95;3 in maize grains was 100%, 93.6% and 90.3%, respectively, and the mean levels were 638.99, 103.54 and 59.69 μg/kg correspondingly. In 12.9% of the maize grain samples, the levels of FBs were higher than the standard reference. The residents were at low exposure risk overall. The mean estimated daily intake(EDI) of FBs was far lower than the provisional maximum tolerable daily intake of 2 μg/(kg·BW·d). However, 0.30% of the residents were at high risk. Among people of different ages, the mean EDI of children, adults, and elderly were 0.43, 0.28 and 0.29 μg/(kg·BW·d) respectively, and children were in the highest exposure levels of FBs. Among the tested five foodstuffs, rice and maize grains were the main sources of FBs exposure., Conclusion: Except for maize grains, the levels of FBs in grains and grain products were relatively low, and Zhejiang residents were at low FBs exposure risk generally.

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.)

This study aims to estimate dietary exposure to deoxynivalenol and fumonisins (FBs) of infants and toddlers in Turkey. A total of 75 processed cereal-based foods intended for infants and toddlers collected between July and October 2018, were analysed by high performance liquid chromatography (HPLC). DON was determined in 21.3% of samples with mean middle bound (MB) level of 28.4 μg/kg. Of the 16 quantifiable samples, only one showed values above 200 μg/kg. Fumonisin B 1 (FB 1 ) was detected at quantifiable levels only in three samples, while FB 2 was not found in any sample. Estimated mean MB chronic dietary exposures to DON in infants and toddlers were 0.161 and 0.118 μg/kg b.w. per day, while 95th percentile (P95) MB exposures to DON were estimated at 0.564 and 0.414 μg/kg b.w. per day, respectively. Mean MB dietary exposures to FBs for infants and toddlers, respectively, were 0.093 and 0.068 μg/kg b.w. per day; P95 exposure estimates were 0.079 and 0.058 μg/kg b.w. per day. Both for DON and FBs, mean and P95 exposures of infants and toddlers did not exceeded the threshold level of 1 μg/kg b.w. per day and are therefore not of health concern., 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 © 2020 Elsevier Ltd. All rights reserved.)

A survey on 120 cereal samples (barley, maize, rice and wheat) from Algerian markets has been carried out to evaluate the presence of 15 mycotoxins (ochratoxin A, deoxynivalenol, fumonisin B1 and B2, T-2 and HT-2 toxins, zearalenone, fusarenon X, citrinin, sterigmatocystin, enniatins A, A1, B and B1, and beauvericin). With this purpose, a QuEChERS-based extraction and ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) were used. Analytical results showed that 78 cereal samples (65%) were contaminated with at least one toxin, while 50% were contaminated with three to nine mycotoxins. T-2 toxin, citrinin, beauvericin and deoxynivalenol were the most commonly found mycotoxins (frequency of 50%, 41.6%, 40.8% and 33.3%, respectively). Fumonisins (B1 + B2), enniatins B and B1, deoxynivalenol and zearalenone registered high concentrations (289-48878 µg/kg, 1.2-5288 µg/kg, 15-4569 µg/kg, 48-2055 µg/kg and 10.4-579 µg/kg, respectively). Furthermore, concentrations higher than those allowed by the European Union (EU) were observed in 21, 8 and 1 samples for fumonisins, zearalenone and deoxinivalenol, respectively. As a conclusion, the high levels of fumonisins (B1 + B2) in maize and deoxynivalenol, zearalenone and HT-2 + T-2 toxins in wheat, represent a health risk for the average adult consumer in Algeria. These results pointed out the necessity of a consistent control and the definition of maximum allowed levels for mycotoxins in Algerian foodstuffs.

Cereal grains are essential ingredient in food, feed and industrial processing. One of the major causes of cereal spoilage and mycotoxin contamination is the presence of toxigenic Fusarium spp. Nanoparticles have immense applications in agriculture, nutrition, medicine or health but their possible impact on the management of toxigenic fungi and mycotoxins have been very little explored. In this report, the potential of silver nanoparticles (AgNPs) (size 14-100 nm) against the major toxigenic Fusarium spp. affecting crops and their effect on mycotoxin accumulation is evaluated for the first time. The studied Fusarium spp. (and associated mycotoxins) were F. graminearum and F. culmorum (deoxynivalenol, 3-acetyldeoxynivalenol and zearalenone), F. sporotrichioides and F. langsethiae (T-2 and HT-2 toxins), F. poae (nivalenol), F. verticillioides and F. proliferatum (fumonisins B 1 and B 2 ) and F. oxysporum (mycotoxins no detected). The factors fungal species, AgNP dose (range 2-45 μg/mL), exposure time (range 2-30 h) and their interactions significantly influence spore viability, lag period and growth rate (GR) in subsequent cultures in maize-based medium (MBM) of all the studied species. The effective lethal doses (ED 50 , ED 90 and ED 100 ) to control spore viability and GR were in the range 1->45 μg/mL depending on the remaining factors. At high exposure times (20-30 h), the three effective doses ranged 1-30 μg/mL for all the studied species. At the end of the incubation period (10 days) mycotoxin levels in MBM cultures inoculated with fungal spores from treatments were strongly related with the size reached by the colony at that time. None of the treatments produced stimulation in conidia germination, GR or mycotoxin biosynthesis with respect to controls. Thus, the antifungal effect of the assayed AgNPs against the tested Fusarium spp. suggests that AgNPs could be a new antifungal ingredient in bioactive polymers (paints, films or coating) likely to be implemented in the agro-food sector for controlling these important toxigenic Fusarium spp. and their main associated mycotoxins., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Maize has become one of the most important crops for food and feed production-both as a silage and crop residue worldwide. The present study aimed to identify the co-occurrence of Fusarium subglutinans , Fusarium verticillioides , Trichoderma atroviride , Sarocladium zeae, and Lecanicillium lecanii on maize ear rot. Further, the accumulation of mycotoxins as secondary metabolites of Fusarium spp. in maize ear samples was also analyzed. Maize ear samples were collected between 2014 and 2017 from two main maize growing areas in Poland (Greater Poland and Silesia region). A significant difference was found in the frequency of two main Fusarium spp. that infect maize ears, namely F. subglutinans and F. verticillioides . In addition to Fusarium spp. T. atroviride , S. zeae, and L. lecanii were also identified. T. atroviride species was found in 14% of maize samples examined between 2014 and 2017, particularly with a high percentage of Trichoderma spp. recorded in 2014, i.e., in 31% of samples. However, mycotoxin content (beauvericin and fumonisins) varied, depending on both the location and year of sampling. The interaction of fungi and insects inhabiting maize ear and kernel is very complex and not yet elucidated. Therefore, further research is required in this area.

Fumonisins are mycotoxins (MTs) produced mainly by the fungus Fusarium verticillioides , the main pathogens of maize which cause ear rot. The aim of this work was to evaluate some factors that may lead to high fumonisin production by F. verticillioides in maize grains, correlating the pathogen inoculation method with different genotypes grown in four Brazilian states. Experiments were conducted in 2015-2016 in maize crops from experimental maize fields located in four distinct states of Brazil. Results showed that contamination by fumonisin mycotoxins occurred even on symptomatic or asymptomatic grains. In all municipalities, the samples showed levels of fumonisin B1 that were higher than would be tolerable for the human consumption of corn products (the current tolerance limit for fumonisin is 1.5 μg g -1 ). High severity of grains infected with F. verticillioides does not always show high concentrations of fumonisins. Environments with higher temperatures may influence the production of high concentrations of fumonisin in maize hybrids. Spray inoculation methods and inoculation at the center of spikes did not influence fumonisin concentrations. Results showed that the hybrids P3630H, P32R48 and P3250 presented higher disease severity, as well as higher mycotoxin levels in the studied locations with higher temperatures.

Consumption of fumonisin-contaminated foods has a negative influence on the health of humans (carcinogen; oesophageal cancer in Eastern Cape in South Africa). Lactic acid bacteria (LAB) have emerged as a promising natural detoxification agent against mycotoxins. The aim of this study was to visualise the interaction between fumonisins (FB 1 and FB 2 ) and LAB: Lactobacillus plantarum FS2, L. delbrueckii subsp. delbrueckii CIP 57.8T and Pediococcus pentosaceus D39, isolated from traditional fermented maize-based products (ogi and mahewu) using confocal laser scanning microscopy (CLSM) and to then quantify the LAB-bound fumonisin using high performance liquid chromatography (HPLC). The objective was to obtain a physically visible and quantifiable binding interaction between fumonisins and LAB strains with the aim of utilising LAB as a possible detoxifying agent. Fumonisins were derivatised using naphthalene-2,3-dicarboxaldehyde (NDA) and then combined with non-fluorescent LAB cells (viable and non-viable). For the quantification of bound fumonisins, viable and non-viable cells were incubated in the presence of predetermined concentrations of fumonisins and the level of fumonisin in the suspension was determined. CLSM showed the derivatised green fluorescent fumonisins binding to the surface of each of the LAB cells. For viable cells, L. plantarum FS2 bound FB 1 most effectively while P. pentosaceus D39 bound the least level of FB 1 . The highest levels of FB 2 were bound by L. plantarum R 1096 and the least by L. delbrueckii CIP 57.8 T. For non-viable cells, L. plantarum FS2 was also the most effective for binding both fumonisins with P. pentosaceus D39 and L. delbrueckii CIP 57.8 T being the least effective for FB 1 and FB 2 , respectively. To our knowledge, this is the first study to visualise the interaction between LAB and fumonisins. We demonstrate that LAB isolates from indigenous fermented maize-based beverages bind fumonisins and thus present a potential strategy for their reduction in these traditional foods.

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.)

Brazil is one of the world's largest corn producers and is a leader in exportation. Due to intense globalization, corn may be commercialized worldwide and the issue concerning the safety of corn-based products has become a topic of widespread international interest. Dietary exposure evaluation is a relevant criterion for mycotoxin risk assessment. Thus, human exposure to fumonisins were assessed for corn grain and its derivatives (endosperm, cornmeal, and grits; n = 320) sampled from one of the large-scale corn processing plants in Brazil. The total probable daily intake (PDI) for fumonisins in Brazil was 96.9 ng kg -1 body weight day -1 , which corresponds to 5% of the provisional maximum tolerable daily intake (PMTDI) of 2000 ng kg -1 b.w. day -1 for fumonisins. In countries that import Brazilian corn, the total PDI is lower in European countries (from 35.7 to 177 ng kg -1 b.w. day -1 ) and higher in Angola (1553 ng kg -1 b.w. day -1 ). Taking into account that dietary exposure in populations in Brazil and importing countries was low, the corn-based products were safe for human consumption regarding fumonisins, even for regions with high corn consumption.

In temperate areas, the main limitation to the use of maize in the food chain is its contamination by B-series fumonisins (FBs) during cultivation. Since the content of this group of mycotoxins may be distributed unevenly after milling, the aim of this study was to compare the distribution of FBs in maize fractions derived from two industrial dry-milling processes, that is, a dry-degermination (DD) system and a tempering-degermination (TD) system. Grain cleaning reduces FBs by about 42%. The germ of the two degermination processes showed a similar FB content of kernel after cleaning. Conversely, an animal feed flour resulted in a FB content that was two times higher than whole grain before cleaning. A significant FB reduction was observed in the milling fractions in both processes, with a higher reduction in the TD system than in the DD one. The average decontamination respect to uncleaned kernels in the DD process was 50%, 83% and 87%, for maize flour, break meal and pearl meal, respectively, while it was 78%, 88% and 94% in the TD process for small, medium and flaking grits, respectively. Among the milling fractions, the flaking grits with the highest particle size resulted in the highest FB reduction.

Background: In the present study, a novel highly sensitive magnetic enzyme chemiluminescence immunoassay (MECLIA) was developed to detect fumonisin B 1 (FB 1 ) in cereal samples. The gold-coated magnetic nanoparticles (Fe 3 O 4 @Au, GoldMag) were used as solid phase carrier to develop a competitive CLIA for detecting FB 1 , in which FB 1 in samples would compete with FB 1 -ovalbumin coated on the surface of Fe 3 O 4 @Au nanoparticles for binding with FB 1 antibodies. Successively, horseradish peroxidase labeled goat anti-rabbit IgG (HRP-IgG) was conjugated with FB 1 antibodies on the microplate. In substrate solution containing luminol and H 2 O 2 , HRP-IgG catalyzed luminol oxidation by H 2 O 2 , generating a high chemiluminescence signal. The FB 1 immune GoldMag particles were characterized by Fourier transform infrared spectroscopy, scanning electron microscope and zeta potential analysis, etc. RESULTS: The concentrations and the reaction times of these immunoreagents were optimized to improve the performances of this method. The established method could detect as low as 0.027 ng mL -1 FB 1 from 0.05 ng mL -1 to 25 ng mL -1 , demonstrating little cross-reaction (less than 2.4%) with other structurally related compounds. The average intrassay relative SD (RSD) (n = 6) was 3.4% and the average interassay RSD (n = 6) was 5.4%. This method was successfully applied for the determination of FB 1 in corn and wheat and gave recoveries of between 98-110% and 91-105%, respectively., Conclusion: The results of the present study suggest that the MECLIA approach has potential application for high-throughput fumonisin screening in cereals. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

A visual immunoaffinity test column (IATC) assay was developed to detect fumonisins in cereal samples for spot tests without the need for special instruments. The developed IATC assay had equivalent recognition capability for fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), or fumonisin B₃ (FB₃), and exhibited no cross-reactivity with aflatoxin B₁, ochratoxin A, zearalenone, or the T-2 toxin. The sample pretreatment was accomplished more rapidly and with greater ease, the entire assay procedure was completed in approximately 10 min, including sample pretreatment and testing. The limits of detection (LODs) of the IATC assay to detect fumonisins in the maize, barley, oat, and millet samples were 20 &mu;g kg &minus;1 . The results of the spiked maize, barley, oat, and millet and real maize samples by the IATC assay agreed well with the results obtained by the commercial fumonisin enzyme-linked immunosorbent assay (ELISA) test kit and liquid chromatography-tandem mass spectrometry (LC-MS/MS), respectively. The developed IATC assay can serve as a useful screening tool for the rapid, qualitative, and semi-quantitative detection of the total content of fumonisins (sum of FB₁, FB₂, and FB₃) in cereal samples on-site.

To assess the risk of fumonisin contamination in Korean cereals, we isolated colonies of the Fusarium fujikuroi species complex (FFSC) from barley, maize, rice and soybean samples from 2011 to 2015. A total of 878 FFSC strains were isolated mostly from maize and rice, and species identity of the isolates were determined using the DNA sequence of the translation elongation factor 1-α (TEF-1α) and RNA polymerase II (RPB2) genes. Fusaria recovered from Korean cereals included F. fujikuroi (317 isolates and a frequency of 36%), F. proliferatum (212 isolates and 24.1%), F. verticillioides (170 isolates and 19.4%), F. concentricum (86 strains and 9.8%), F. andiyazi (56 isolates and 6.4%), F. subglutinans (28 isolates and 3.2%), F. thapsinum (5 isolates and 0.6%), and F. circinatum (2 isolates and 0.2%). The rice samples were dominated by F. fujikuroi (47.4%), F. proliferatum (27.3%), and F. concentricum (15.1%), whereas maize samples were dominated by F. verticillioides (33.9%), F. fujikuroi (25.3%), and F. proliferatum (21.1%). A phylogenetic analysis of 70 representative isolates demonstrated that each species was resolved as genealogically exclusive in the ML tree. Fumonisin production potential was evaluated using a PCR assay for the fumonisin biosynthesis gene, FUM1 in all of the isolates. Most of the isolates tested (94%) were positive for FUM1. All of the isolates assigned to F. fujikuroi, F. proliferatum, F. verticillioides and F. thapsinum were positive for FUM1 irrespective of their host origin. Seventy-seven representative isolates positive for FUM1 were examined for fumonisin production in rice medium. The majority of F. proliferatum (26/27, 96.3%), F. verticillioides (16/17, 94.1%) and F. fujikuroi (19/25, 76.0%) produced both FB 1 and FB 2 . Notably, 16 of 19 fumonisin-producing F. fujikuroi produced >1000μg/g of fumonisins (FB 1 +FB 2 ) in rice medium, which is higher than that in previous reports. These results suggest that F. fujikuroi can produce high levels of fumonisins similar to F. verticillioides and F. proliferatum., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A fast, easy, and cheap method for the simultaneous determination and quantification of aflatoxins (B1, B2, G1, G2), T-2 and HT-2 toxins, and fumonisins (B1, B2) in cereal-derived products was developed. This method involved a quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction coupled with liquid chromatography-tandem mass spectrometry. The method validation was performed by analyzing samples spiked at four levels, and the recoveries ranged from 83.6 to 102.9%, whereas the maximum values of repeatability and within-laboratory reproducibility were 14.3 and 15.7% following the performance criteria set by the European legislation. The method was then applied for the analysis of 21 cereal-derived products purchased on the Italian market, which were correctly packaged and labeled as intended for human consumption. The co-occurrence of more than one mycotoxin in the analyzed samples could represent a risk for consumers, and the described method could be a valid alternative for their simultaneous detection in the framework of official control. Graphical Abstract ᅟ.

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.)

Mycotoxin contamination in rice is generally lower than in other cereals such as corn or wheat. However, over 65% of the population in China consumes rice as a staple food. Due to the diversity of the climate across China, the southern region is characterized by high temperatures and humidities, especially in rainy season. Such conditions are optimal for the growth of fungi. The accumulative and transferrable characteristics of fungi mycotoxins pose a great potential threat as confirmed by high incidences of liver cancer in the Yangtze delta region. Major mycotoxins identified in China are aflatoxins and ochratoxin A, as well as fumonisins. The contents of aflatoxin B 1 (AFB 1 ) in rice are varied among different provinces and regions and generally less than 5 μg/kg. Although high incidences of positive aflatoxins samples have widely been detected, few samples were detected as exceeding the national's maximum residue limit (10 μg/kg). Limited information is available on risk assessment of human health hazards of mycotoxins in rice, children should be paid more attention to due to their having the highest mycotoxins exposure level, although the risks are generally at low levels from rice. Mycotoxins are mainly distributed in the outer layer of the paddy rice (also called rough rice, referring to whole rice grain with the hulls), and the AFB 1 content in bran is 8.4 times greater than that in brown rice (hulled rice). Further investigation should focus on isolation and identification of mycotoxins-producing fungal strains, especially unknown mycotoxigenic fungal strains determination. Infection resistant rice breeding of mycotoxigenic fungal species may be a fundamental approach to guaranteeing rice safety in China., (© 2017 Institute of Food Technologists®.)

In vitro experimental environments are used to study interactions between microorganisms, and to predict dynamics in natural ecosystems. This study highlights that experimental in vitro environments should be selected to match closely the natural environment of interest during in vitro studies to strengthen extrapolations about aflatoxin production by Aspergillus and competing organisms. Fungal competition and aflatoxin accumulation were studied in soil, cotton wool or tube (water-only) environments, for Aspergillus flavus competition with Penicillium purpurogenum, Fusarium oxysporum or Sarocladium zeae within maize grains. Inoculated grains were incubated in each environment at two temperature regimes (25 and 30°C). Competition experiments showed interaction between the main effects of aflatoxin accumulation and the environment at 25°C, but not so at 30°C. However, competition experiments showed fungal populations were always interacting with their environments. Fungal survival differed after the 72-h incubation in different experimental environments. Whereas all fungi incubated within the soil environment survived, in the cotton wool environment none of the competitors of A. flavus survived at 30°C. With aflatoxin accumulation, F. oxysporum was the only fungus able to interdict aflatoxin production at both temperatures. This occurred only in the soil environment and fumonisins accumulated instead. Smallholder farmers in developing countries face serious mycotoxin contamination of their grains, and soil is a natural reservoir for the associated fungal propagules, and a drying and storage surface for grains on these farms. Studying fungal dynamics in the soil environment and other environments in vitro can provide insights into aflatoxin accumulation post-harvest.

Ultrahigh-performance liquid chromatography using positive electrospray ionization and quadrupole orbital ion trap high-resolution mass spectrometry was evaluated for analyzing mycotoxins in finished cereal and nut products. Optimizing the orbital ion trap mass analyzer in full-scan mode using mycotoxin-fortified matrix extracts gave mass accuracies, δM, of < ± 2.0 ppm at 70,000 full width at half maximum (FWHM) mass resolution (RFWHM). The limits of quantitation were matrix- and mycotoxin-dependent, ranging from 0.02 to 11.6 μg/kg. Mean recoveries and standard deviations for mycotoxins from acetonitrile/water extraction at their relevant fortification levels were 91 ± 10, 94 ± 10, 98 ± 12, 91 ± 13, 99 ± 15, and 93 ± 17% for corn, rice, wheat, almond, peanut, and pistachio, respectively. Nineteen mycotoxins with concentrations ranging from 0.3 (aflatoxin B1 in peanut and almond) to 1175 μg/kg (fumonisin B1 in corn flour) were found in 35 of the 70 commercial grain and nut samples surveyed. Mycotoxins could be identified at δM < ± 5 ppm by identifying the precursor and product ions in full-scan MS and data-dependent MS/MS modes. This method demonstrates a new analytical approach for monitoring mycotoxins in finished grain and nut products.

An immunochromatographic strip (ICS) using urchin-like gold nanoparticles (UGNs) for sensitive detection of fumonisin B1 (FB1) was developed to meet the requirement for rapidly monitoring FB1 in grain samples. The sensitivity of the ICS was 5.0 ng/mL, which represents a fourfold increase in sensitivity over conventional strip preparation using colloidal gold as the antibody-labeled probe. Analysis of FB1 in grain samples showed that data obtained from the strip tests were in a good agreement with those obtained from HPLC and enzyme-linked immunosorbent assays (ELISAs). This qualitative test did not require any specialized equipment, and the detection time was less than 5 min, which is suitable for on-site testing of FB1 in grain samples. Overall, to our knowledge, this is the first report of using a UGN as the antibody-labeled probe for sensitive detection of FB1 in grains using an ICS. Graphical Abstract Preparation of ICS using conventional colloidal gold and urchin-like gold nanoparticle, respectively.

Mycotoxins including aflatoxins, deoxynivalenol, fumonisins and ochratoxin A are among the main fungal secondary metabolites detected as natural contaminants in South America in different commodities such as peanuts (aflatoxins), cereals (deoxynivalenol and fumonisins) or grapes (ochratoxin A). Different strategies including crop rotation, tillage practices, fungicide application and planting less susceptible cultivars are used in order to reduce the impact of these mycotoxins in both food and feed chains. The development of fungicide resistance in many fungal pathogens as well as rising of public concern on the risks associated with pesticide use led to the search for alternative environmentally friendly methods. Biological control of plant pathogens and toxigenic fungi offers an alternative that can complement chemical control in the frame of an integrated pest management to reduce the impact of mycotoxins in the food and feed chains. The advances made in Argentina on reducing the impact of toxigenic fungi and mycotoxins in peanut, grapes and cereals using the biocontrol strategy are summarised. Native bacteria, yeasts and filamentous fungi have been selected to evaluate them as potential biocontrol agents. Field trials showed that Bacillus subtilis RC 218 and Brevibacillus sp. RC 263 were effective at reducing deoxynivalenol accumulation in wheat. The application of Clonostachys rosea isolates on wheat stubble reduced Fusarium colonisation on the stubble. Bacillus amyloliquefaciens and Microbacterium oleovorans showed good activity to control both Fusarium verticillioides growth and the accumulation of fumonisins at pre-harvest stage in maize. Control of toxigenic Aspergillus flavus and aflatoxin accumulation in peanuts was achieved using a native atoxigenic Aspergillus flavus strain based on competitive exclusion of the toxigenic strains. Kluyveromyces thermotolerans strains were used as biocontrol agents to reduce the impact of Aspergillus section Nigri and ochratoxin A accumulation in grapes.

Mycotoxins that commonly occur in cereal grains and other products can contaminate finished processed foods on account of their high toxicity. The mycotoxins that are commonly associated with food grains include aflatoxins, ochratoxin A, fumonisins, deoxynivalenol, and zearalenone. Various food-processing operations include sorting, trimming, cleaning, cooking, baking, frying, roasting, flaking, and extrusion that have variable effects on mycotoxins. The nature of the processing operation viz. physical, chemical, or thermal plays an important role in this; usually, the processes that utilize the higher temperatures have greater effects on mycotoxin dissipation. In general, the processes are known to reduce mycotoxin concentrations significantly, but do not eliminate them completely. However, roasting and extrusion processing result in lowest mycotoxin concentrations, since these involve higher temperatures. It is observed that very high temperatures are needed to bring about high reduction in mycotoxin concentrations, approaching acceptable background levels. The treatment with chemicals like ammonia, bicarbonate, citric acid, or sodium bisulfite is also effective in resulting in significant decline in mycotoxin concentrations.

In this study a total of 522 samples were collected from Shandong province of China in 2014 and analysed for the occurrence of fumonisin B1 (FB1), FB2 and FB3 by isotope dilution ultrahigh performance liquid chromatography-tandem mass spectrometry. Fumonisins were detected in 98.1% of the corn products, with the average total level of 369.2 μg kg(-1). The individual average values of FB1, FB2 and FB3 in corn products were 268.3, 53.7 and 47.2 μg kg(-1), respectively. The simultaneous occurrence of FB1, FB2 and FB3 was observed in 76.7% of the corn products. Especially, the results demonstrated that the difference in the contamination levels for fumonisins in these three types of corn products was apparent. In addition, 6.2% of the wheat flour samples were contaminated with FB1, with concentrations ranging from 0.3 to 34.6 µg kg(-1). No FB2 or FB3 was detected in wheat flour. In corn oil samples no fumonisins were detected.

A simple, new aptamer-photonic crystal encoded suspension array was designed to simultaneously quantify and qualify ochratoxin A(OTA) and fumonisin B1(FB1) in cereal samples. The aptamers of OTA and FB1 were immobilized on the surfaces of photonic crystals by chemical bonding. When the target mycotoxins appear in a sample, the fluorescence-labeled complementary DNA of the aptamer dissociates from their double DNA hybrid and results in an obvious decrease in fluorescence intensity of the microsphere. The difference value of fluorescent intensities for each kind of silica photonic crystal microsphere (SPCM) quantitatively conveys the concentration of mycotoxin, and the structure colors or reflectance peak positions of the SPCMs confirm the kind of mycotoxin detected. The reaction conditions including the immobilization method for aptamers, hybridization, and incubation conditions have been optimized. This developed method displayed a wide linear detection range (0.01-1 ng/mL for OTA and 0.001-1 ng/mL for FB1) and a low limit of detection (0.25 pg/mL for OTA and 0.16 pg/mL for FB1). The recovery rates in the spiked cereal samples ranged from 81.80% to 116.38% for OTA and 76.58%-114.79% for FB1. The positive detection results in the naturally contaminated cereal samples were in agreement with those of classic enzyme-linked immunosorbent assay (ELISA). This simple suspension array scheme displays a great application potential for the high throughput screen assay of mycotoxins.

The aim of this study was to detect and quantify fumonisin B1 (FB1) in cereal mixtures marketed in Brazil. Fifteen samples from different lots were acquired each month by internet from supermarkets during seven months, adding up to 105 analysed samples. The unit sample constituted of an original package with a minimum of 250 g. Extraction and clean-up of samples for FB1 determination were carried out using immunoaffinity columns. Identification and quantification of FB1 were performed by high performance liquid chromatography. Eighty-eight (83.8%) samples were contaminated with FB1 and four (3.8%) presented levels above 500 µg kg(-1) (634, 703, 1269 and 1876 µg kg(-1)). Maximum FB1 + FB2 levels allowed by Brazilian regulations will reach 1500 µg kg(-1) for corn flour in 2016 and 1000 µg kg(-1) for others corn products. This study showed that even at levels below the legislative limits, human exposure to this toxin can occur constantly.

A combined (triplex) immunoassay for the simultaneous detection of three mycotoxins in grains was developed with superparamagnetic colour-encoded microbeads, in combination with two bead-dedicated flow cytometers. Monoclonal antibodies were coupled to the beads, and the amounts of bound mycotoxins were inversely related to the amounts of bound fluorescent labelled mycotoxins (inhibition immunoassay format). The selected monoclonal antibodies were tested for their target mycotoxins and for cross-reactivity with relevant metabolites and masked mycotoxins. In the triplex format, low levels of cross-interactions between the assays occurred at irrelevant high levels only. All three assays were influenced by the sample matrix of cereal extracts to some extent, and matrix-matched calibrations are recommended for quantitative screening purposes. In a preliminary in-house validation, the triplex assay was found to be reproducible, sensitive and sufficiently accurate for the quantitative screening at ML level. The triplex assay was critically compared to liquid chromatography-tandem mass spectrometry using reference materials and fortified blank material. Results for the quantification of ochratoxin A and zearalenone were in good agreement. However, the fumonisin assay was, due to overestimation, only suitable for qualitative judgements. Both flow cytometer platforms (Luminex 100 and FLEXMAP 3D) performed similar with respect to sensitivity with the advantages of a higher sample throughput and response range of the FLEXMAP 3D and lower cost of the Luminex 100.

A fast and effective method for quantitative determination of fumonisins B1 and B2 (FB1 and FB2) in different cereal-based products was developed. Fumonisins were extracted combining a solid liquid extraction (SLE) pre-treatment and immunoaffinity columns (IACs), which were applied for an effective clean-up. Consecutively, high pressure liquid chromatography (HPLC) coupled to hybrid triple quadrupole-linear ion trap mass spectrometer (QTrap®) was used for the separation, detection and quantification of targeted mycotoxins. The analytical method showed acceptable recoveries ranged from 89.7% to 99.1%, as well as a relative standard deviation (%, RSD) lower than 12%. A total of 1250 samples from France, Germany and Spain were analysed. The incidence of fumonisin B1 and B2 in organic samples were 11.4% and 11.3%, respectively. By contrast, the occurrence of conventional samples was 3.6% of FB1 and 3.5% of FB2. The highest values were obtained in Spanish samples of flour, as an example toasted corn, called gofio that was contaminated with 1201.7 and 1010.5 μg kg(-1) of FB1 and FB2, respectively., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Monitoring results of food grain contamination with fusariotoxins-deoxynivalenol (DON), zearalenone (ZEN), fumonisins (FB1&FB2), T-2 and HT-2 toxins-are presented. Harvests of 2005-2010 in different regions of Russia were investigated. The occurrence of DON in wheat was 8%, barley 9%, oats 4%, rye 2% and maize 2%. The highest frequency of ZEN contamination was found in oats, the lowest in wheat. Calculated average daily intake of DON varied from 0.066 to 0.096 µg/kg body weight, the highest being found in the Southern region, but substantially lower than the provisional maximum tolerable daily intake. The results of enzyme-linked immunosorbent assay and high-performance liquid chromatography-mass spectrometry analysis demonstrated the presence of T-2 toxin in 14% and HT-2 toxin in 17% of all samples. The maximum level of T-2 toxin was exceeded only in one sample of barley. Relatively high frequency and levels of FB1&FB2 contamination were found in maize.

Distiller's dried grains with solubles (DDGS) is a major coproduct of the fuel-ethanol industry and is becoming a popular low-cost ingredient for animal feed. Uncertainties regarding the risk factors in DDGS, such as level of mycotoxins, could limit its application in the animal feed industry. To provide a scientifically sound assessment of the prevalence and levels of mycotoxins in U.S. DDGS, we measured aflatoxins, deoxynivalenol, fumonisins, T-2 toxin, and zearalenone in 67 DDGS samples collected from 8 ethanol plants in the midwestern United States from 2009 to 2011. Among the five mycotoxins, deoxynivalenol was the main focus of the study because the crop year of 2009 was favorable for deoxynivalenol occurrence in corn. We learned that no more than 12% of the samples contained deoxynivalenol levels higher than the minimum advisory level for use in animal feed provided by the U.S. FDA, and the deoxynivalenol levels in all DDGS collected in 2011 were <2 mg/kg. Besides, intensive study showed that the enrichment of deoxynivalenol from contaminated corn to DDGS was about 3.5 times. With regard to the other mycotoxins in DDGS, the study suggested that (1) almost none of the DDGS samples produced in 2010 contained detectable aflatoxins and the highest level of aflatoxins in DDGS was 5.7 μg/kg; (2) no more than 6% of the samples contained fumonisin levels higher than the guidance level for feeding equids and rabbits provided by the U.S. FDA; (3) none of the samples contained T-2 higher than the detection limit; (4) most samples contained zearalenone levels between 100 and 300 μg/kg. This study was based on representative DDGS samples from the U.S. ethanol industry, and the data were collected using state-of-the-art analytical methodology. This study provided a comprehensive and scientifically sound assessment of the occurrence and levels of mycotoxins in DDGS produced from 2009 to early 2011 by the U.S. ethanol industry.

Introduction: Fumonisin B1 (FB1) is a toxic and carcinogenic mycotoxin produced in cereals due to fungal infection. This study was conducted to determine FB1 contamination of rice and corn samples and its relationship with the rate of esophageal cancer (EC) in a high risk area in northeastern Iran., Methods: In total, 66 rice and 66 corn samples were collected from 22 geographical subdivisions of Golestan province of Iran. The levels of FB1 were measured for each subdivision by thin layer and high pressure liquid chromatographies. The mean level of FB1 and the proportions of FB1 contaminated samples were compared between low and high EC-risk areas of the province., Results: The mean of FB1 levels in corn and rice samples were 223.6 and 21.6 μg/g, respectively. FB1 contamination was found in 50% and 40.9% of corn and rice samples, respectively. FB1 level was significantly higher in rice samples obtained from high EC-risk area (43.8 μg/g) than those obtained from low risk area (8.93 μg/g) (p-value=0.01). The proportion of FB1 contaminated rice samples was also significantly greater in high (75%) than low (21.4%) EC-risk areas (p-value=0.02)., Conclusion: We found high levels of FB1 contamination in corn and rice samples from Golestan province of Iran, with a significant positive relationship between FB1 contamination in rice and the risk of EC. Therefore, fumonisin contamination in commonly used staple foods, especially rice, may be considered as a potential risk factor for EC in this high risk region.

An LC-MS/MS method was developed and validated for the simultaneous determination of deoxynivalenol, zearalenone, T-2-toxin, HT-2-toxin and metabolites, including 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deoxynivalenol-3-glucoside, α-zearalenol, β-zearalenol, zearalenone-4-glucoside, α-zearalenol-4-glucoside, β-zearalenol-4-glucoside and zearalenone-4-sulfate in maize, wheat, oats, cornflakes and bread. Extraction was performed with acetonitrile/water/acetic acid (79/20/1, v/v/v) followed by a hexane defatting step. After filtration, the extract was evaporated and the residue was redissolved in mobile phase for injection. The mobile phase, which consisted of a mixture of methanol and water with 10 mM ammonium acetate, was adjusted to pH 3 with glacial acetic acid. A sample clean-up procedure was not included because of the low recoveries of free and masked mycotoxins and their differences in polarity. The method allowed the simultaneous determination of 13 Fusarium mycotoxins in a one-step chromatographic run using a Waters Acquity UPLC system coupled to a Quattro Premier XE mass spectrometer. The method was validated for several parameters such as linearity, apparent recovery, limit of detection, limit of quantification, precision, expanded measurement uncertainty and specificity. The limits of detection varied from 5 to 13 ng g⁻¹; those for the limit of quantification from 10 to 26 ng g⁻¹. The results of the performance characteristics of the developed LC-MS/MS method were in good agreement with the criteria mentioned in Commission Regulation (EC) No. 401/2006. Thirty samples of a variety of food and feed matrices were sampled and analysed between July 2010 and January 2011.

Background: The genus Fusarium comprises a diverse group of fungi including several species that produce mycotoxins in food commodities. In the present study, a multiplex PCR was standardised for the group-specific detection of fumonisin-producing and trichothecene-producing strains of Fusarium species. Primers for genus-level recognition of Fusarium spp. were designed from the internal transcribed spacer regions 1 and 2 of rDNA. Primers for group-specific detection were designed from the tri5 and tri6 genes involved in trichothecene biosynthesis and the fum1 and fum13 genes involved in fumonisin biosynthesis., Results: Among the various genera and their strains tested, all the 85 confirmed Fusarium strains were positive for rDNA gene and the rest stayed negative. From among the Fusarium strains, 15 had amplification for trichothecene- and 20 for fumonisin-encoding genes. All PCR positive trichothecene chemotypes of Fusarium species tested were positive for chemical analysis but in the case of fumonisins, of the 20 PCR positive cultures, only 13 showed positive for chemical analysis by HPTLC., Conclusion: The assay described here provided a rapid and reliable detection of trichothecene- and fumonisin-producing Fusarium directly from natural food grains and the results were always comparable with a conventional HPTLC detection method. It can, therefore, be used by the food industry to monitor quality and safety., (Copyright © 2011 Society of Chemical Industry.)

A comparison study of different extraction and clean-up procedures for the liquid chromatographic analysis of fumonisins B(1) (FB(1)) and B(2) (FB(2)) in corn masa flour was performed. The procedures included extraction (heat or room temperature) with acidic conditions or EDTA-containing solvents, and clean-up by immunoaffinity or C18 solid-phase extraction columns. Thereafter an analytical method was optimised using extraction with an acidic mixture of methanol-acetonitrile-citrate/phosphate buffer, clean-up through the immunoaffinity column and determination of fumonisins by liquid chromatography with automated pre-column derivatisation with o-phthaldialdehyde reagent. Recovery experiments performed on yellow, white and blue masa flours at spiking levels of 400, 800 and 1200 µg kg(-1) FB(1) and of 100, 200 and 300 µg kg(-1) FB(2) gave overall mean recoveries of 99% (±6%) for FB(1) and 88% (±6%) for FB(2). Good recoveries (higher than 90% for both FB(1) and FB(2)) were also obtained with corn tortilla chips. The limits of quantification of the method (signal-to-noise ratio of 10) were 25 µg kg(-1) for FB(1) and 17 µg kg(-1) for FB(2). The method was tested on different commercial corn masa flours as well as on white and yellow corn tortilla chips, showing fumonisin contamination levels (FB(1) + FB(2)) up to 1800 µg kg(-1) (FB(1) + FB(2)) in masa flour and 960 µg kg(-1) in tortilla chips. Over 30% of masa flours originating from Mexico exceeded the European Union maximum permitted level.

Background: Maize contaminated with mycotoxin fumonisin B1 poses a global threat to agricultural production. In this study, polyclonal antibodies (pAb) specific to fumonisin B1 were generated from rabbits immunised with fumonisin B1-keyhole limpet haemocyanin (KLH). These antibodies were used to establish a sensitive competitive direct enzyme-linked immunosorbent assay (cdELISA) and gold nanoparticle immunochromatographic strip for detecting fumonisin B1 levels in maize-based foods and feeds., Results: In cdELISA, fumonisins B1, B2 and B3 at concentrations of 0.42, 0.58 and 81.5 ng mL(-1) respectively caused 50% inhibition (IC(50)) of binding of fumonisin B1-horseradish peroxidase (HRP) to the antibodies. Effective on-site detection of fumonisin B1 was achieved by developing a rapid and sensitive pAb-based gold nanoparticle immunochromatographic strip. This strip had a detection limit of 5 ng mL(-1) for fumonisin B1 in maize-based samples. Additionally, the whole analytical process could be completed within 10 min. Close examination of 15 maize-based samples by cdELISA revealed that 11 were fumonisin-positive, with a mean concentration of 435 +/- 20.1 ng g(-1). These results correlated well with those obtained by immunochromatographic strip., Conclusion: Both cdELISA and immunochromatographic strip methods established in this study are sensitive for rapid detection of fumonisins in agricultural commodities.

Aspergillus, Penicillium, and Fusarium species frequently contaminate crops. For this reason mycotoxins such as aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FBs), and zearalenone (ZEA) are found in food and feed in a wide range of concentrations, depending on environmental and storage conditions. Consumption of mycotoxin-contaminated food and feed has been associated with acute and chronic poisoning and carcinoma. The aim of this study was to determine the incidence and co-occurrence of AFs (B1+B2+G1+G2), OTA, FBs (B1+B2+B3), and ZEA in 37 samples of cereals and feed randomly collected in 2007 from households of an endemic nephropathy (EN) area in Croatia. The mycotoxins were determined using the competitive direct ELISA test (CD-ELISA) in combination with thin-layer chromatography (TLC). The most frequent mycotoxin was ZEA (92%, mean 318.3 microg kg-1), followed by FBs (27%, 3690 microg kg-1), AFs (24.3%, 4.6 microg kg-1), and OTA (16.2%, 9.8 microg kg-1). Levels of AFs, ZEA, and FBs detected by CD-ELISA significantly correlated with the TLC results. However, only one OTA-positive sample was confirmed by TLC due to its high limit of detection. The levels of these mycotoxins were below the permissible limit for animal feed. Twenty-nine percent of cereals were contaminated with FBs, OTA, or ZEA in mass fractions above the permissible limit for humans. Co-occurrence of two toxins varied between 4.2% and 54% and of three between 4.2% and 7.6%. Prolonged co-exposure to AFs, OTA, FBs, and ZEA might increase the risk of various chronic diseases.

The objective was to determine the effect of added sodium chloride, barley malt and sucrose on the stability of fumonisin B1 (FB1) present in corn flour. Two levels of both sodium chloride (0.4% and 2%) and barley malt (0.8% and 5%) were added to the unextruded corn flour, and six levels of sucrose (3-10%) were used. The addition of sucrose at the lowest salt content (0.4%) as well as salt, either at 0.4% or at 2%, led to a significant decrease of FB1 levels in extruded samples, whereas malt, either at 0.8% or at 5%, did not significantly affect FB1 stability. Decontamination rates depended on the concentrations of added ingredients and ranged from 2% to 92%. The greatest reductions in FB1 content were achieved with extrusion cooking with a high salt content, whilst the lowest reductions were the result of processing corn flour with low contents of both salt and sucrose. Salt at 2% was the most effective ingredient in reducing FB1 content of the final extruded food.

One hundred and fifty-six samples of breakfast cereals were collected from the Canadian retail marketplace over a 3-year period. The samples were analysed for the mycotoxins deoxynivalenol, nivalenol, HT-2 toxin, zearalenone, ochratoxin A, and fumonisins B1 and B2 to contribute to dietary exposure estimates in support of the development of Canadian guidelines for selected mycotoxins in foods. The samples included corn-, oat-, wheat- and rice-based cereals, as well as mixed-grain cereals, and were primarily from North American processors. Overall, deoxynivalenol was the most frequently detected mycotoxin--it was detected in over 40% of all samples analysed. Fumonisins and ochratoxin A were each detected in over 30% of all samples. Zearalenone was detected in over 20% of all samples. Nivalenol and HT-2 toxin were each detected in only one sample. The survey clearly demonstrated regular occurrence of low levels of multiple mycotoxins in breakfast cereals on the Canadian market.

Fumonisin B1 (FB1) is reportedly the causative agent of several animal mycotoxicoses and has etiologically been linked to human oesophageal and liver cancer in certain areas of South Africa and China. To study a possible relationship between exposure to FB1 and human cancer risk, the current status of FB1 contamination in food samples in Huaian and Fusui, where incidences of oesophageal and liver cancer are amongst the highest in China, was investigated. A total of 259 corn samples were collected from individual households in five villages of different townships in Huaian during December 2001 and December 2002, and in four villages of different townships in Fusui during May 2001 and May 2002. Corn samples were also collected from individual households in two villages in Huantai, an area with low incidences of both cancers. Enzyme-linked immunosorbent assays (ELISA) and immunoaffinity-HPLC methods were used for FB1 analysis. In corn samples from Huaian, FB1 was detectable in 95.7% (112/117) of the samples, with an average of 2.84 mg kg-1 (range 0.1-25.5 mg kg-1). FB1 was detected in 83.0% (78/94) of the Fusui samples, with an average of 1.27 mg kg-1 (range 0.1-14.9 mg kg-1), and in 83.3% (40/48) of Huantai samples, with an average of 0.65 mg kg-1 ranging from 0.1 to 5.7 mg kg-1. The level of FB1 in corn samples from Huaian was significantly higher than from Huantai (P < 0.001). In addition, 47 of 112 (42.0%) positive Huaian samples had FB1 level greater than 2.0 mg kg-1, which was significantly higher than 10.0% (4/40) of Huantai samples (P < 0.001). Furthermore, variations were found between samples collected in different years and different villages. The high contamination rates of FB1 found in food from these areas, along with previous reports, suggest a possible contributing role of FB1 in human esophageal- and hepato-carcinogenesis.

This survey examined 30 samples of rice (n = 10), maize (n = 10) and peanuts (n = 10) from Côte d'Ivoire for aflatoxin B1, fumonisin B1 and zearalenone using immunoassays, and ochratoxin A using a validated HPLC method with fluorescence detection. In Côte d'Ivoire, as in other countries, several mycotoxins are present in the same commodities. These mycotoxins are from different structural families: aflatoxin B1, fumonisin B1, zearalenone and ochratoxin A, normally produced by fungal species from Aspergillus, Penicillium and Fusarium genera. Some samples contained four mycotoxins (86%). Four peanuts samples did not show ochratoxin A (14%), whereas they contained aflatoxin B1 concentrations above the EU regulatory limits. Concentrations of ochratoxin A, zearalenone and fumonisin B1 were low and may not cause problems per se; however, fears remain that the tolerable daily intake may be exceeded due to eating habits and synergistic effects could be important with the combination of several mycotoxins. Investigations in this direction are underway, together with isolation and characterization of the fungal species involved.

The occurrence of mycotoxins in barley, sorghum, teff (Eragrostis tef) and wheat from Ethiopia has been studied. Samples were analyzed for aflatoxin B(1) (AFB1), ochratoxin A (OTA), deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN) using high performance liquid chromatography (HPLC) and for fumonisins (FUM) using enzyme linked immunosorbent assay (ELISA). AFB1 and OTA were detected in samples of all the four crops. AFB1 was detected in 8.8% of the 352 samples analyzed at concentrations ranging from trace to 26 microg kg(-1). OTA occurred in 24.3% of 321 samples at a mean concentration of 54.1 microg kg(-1) and a maximum of 2106 microg kg(-1). DON occurred in barley, sorghum and wheat at 40-2340 microg kg(-1) with an overall incidence of 48.8% among the 84 mainly 'suspect' samples analyzed; NIV was co-analyzed with DON and was detected at 40 microg kg(-1) in a wheat sample and at 50, 380, and 490 microg kg(-1) in three sorghum samples. FUM and ZEN occurred only in sorghum samples with low frequencies at concentrations reaching 2117 and 32 microg kg(-1), respectively. The analytical results indicate higher mycotoxin contamination in sorghum, which could be related to the widespread storage of sorghum grain in underground pits leading to elevated seed moisture contents. This is the first report on the occurrence of OTA in teff.

Results from the Bavarian Health and Food Safety Authority on contamination of cereals and cereal products from the Bavarian market with the mycotoxins deoxynivalenol (DON), zearalenone (ZEA), and ochratoxin A (OTA) and of maize meal and semolina with fumonisins (FUM) in the year 2004 are presented. Contamination rates and levels of DON, ZEA, and OTA were low and did not exceed the maximum levels. However, a 92% contamination rate and high levels of FUM in maize meal and semolina were measured. Contamination levels of mycotoxins are discussed and evaluated with respect to possible health implications for consumers.

Objective: To detect the contamination level of fumonisin B1 in grain, and to develop rapid detection kit that possess patent of China., Methods: Hybridoma cell line excreting monoclonal antibody against fumonisin B, was produced using B cell hybridoma technique and develop a rapid, sensitive, quantitative ELISA-kit for detection fumonisin B1., Results: The monoclonal antibody used in the ELISA-kit were tested for subtype as IgG1 and its affinity constant was 8.3 x 10(-8) mol/L. The monoclonal antibody obtained in the present study was highly specific to fumonisin B1 , because no cross reactions between the monoclonal antibodies against fumonisin B1 with the analogues of fumonisin B, were found. The limited concentration of the ELISA-kit was 5 ng/ml, linear range was 50-500 ng/ml, the linear equation was Y = -0.582 x +1.793( = 0.99, P < 0.05). The recovery rate of maize on the level of 50n g/ml, 200 ng/ml and 500 ng/ml was among 71.89%-112.95%. The kit can be stored at normal tempertature in ten months at least. The coefficient of variant winthin-laboratory and between-laboratory was less than 20%.

Brewing and consumption of traditional beer have social-economic significance in most African countries including Botswana. Traditional sorghum malt, wort, and beer samples were collected from three villages around Gaborone, Botswana. Forty-six malt samples were analyzed for fungi on three different media and developing colonies were subcultured for identification. Rhizopus, Fusarium, Mucor, and Aspergillus were the most common genera isolated. Out of the 46 malt samples, 72% contained Rhizopus stolonifer, 63% Fusarium verticillioides (syn. Fusarium moniliforme), and 37% Aspergillus flavus. Although Aspergillus flavus was isolated from malt samples, aflatoxins (B(1), B(2), G(1), and G(2)) were not detected in any of the samples analyzed. When the malt, wort, and beer samples were analyzed for fumonisin B(l) and zearalenone, fumonisin B(1) was detected in 3 malt samples, with concentrations ranging from 47 to 1316 mug/kg, while zearalenone was detected in 56%, 48% and 48% of the malt, wort and beer samples, respectively. Zearalenone concentration in samples ranged from 102 to 2213 mug/kg in malt, 26 to 285 mug/l in wort and 20 to 201 mug/l, in beer. Zearalenone carry-over from wort to beer ranged from 23 to 403%. Therefore, although aflatoxins and fumonisin B(1) do not appear to be major contaminants, zearalenone is common and could pose a potential problem in traditional beer in Botswana.

Extrusion cooking is one of the fastest growing food-processing operations in recent years due to several advantages over traditional methods. Apart from its main goal of improving the quality of intermediate and final processed products, it may incidentally also improve safety because of the potential to reduce mycotoxin levels in cereals. This review is focused on extrusion cooking and aims to give a general overview of its impact in reducing mycotoxin levels in cereals. Extrusion cooking generally decreases the mycotoxins levels at rates depending on different factors such as the type of extruder, the type of screw, the die configuration, the initial mycotoxin concentration, the barrel temperature, the screw speed, the moisture content of the raw material and the use of additives. Reductions of 100, 95 and 83% for fumonisins, aflatoxins and zearalenone, respectively, have been reported during extrusion cooking of cereals, while lower reductions were observed for deoxynivalenol, ochratoxin A and moniliformin, where maximum reductions did not exceed 55, 40 and 30%, respectively.

A survey was performed to obtain the frequency and levels of contamination by deoxynivalenol (DON) and fumonisins B1 and B2 (FB1, FB2) mycotoxins in Italian marketed foods. Of 202 samples investigated, including raw materials and processed cereal foods (bread, pasta, breakfast cereals, biscuits, baby and infant foods), 84% were contaminated with DON at levels from 0.007 to 0.930 microg x g(-1) (median 0.065 microg x g(-1)); 26% contained FB1 ranging from 0.010 to 2.870 microg x g(-1) (0.070 microg x g(-1)); 35% contained FB2 at 0.010-0.790 microg x g(-1) (0.080 microg x g(-1)). The highest levels of DON and FB1 were detected in raw cereals and wholemeal flours. The highest levels of FB2 were detected in durum wheat pasta. A widespread DON contamination was found in baby and infant foods at levels varying from 0.007 to 0.166 microg x g(-1).

Successful use of fluorescence polarization assays (FPAs) in human clinical, infectious disease, and drug discovery fields has prompted us to extend its use to the grain mycotoxin field. An antibody specific to a mycotoxin and a mycotoxin-fluorophore conjugate are developed. Free toxin (extracted from the grains with a suitable solvent) competes with the toxin-fluorophore conjugate for the antibody and a change in FP relative to the quantity of free toxin occurs. This change is compared to a standard curve obtained by using known quantities of toxin. The use of FP and toxin-fluorophore conjugates for the quantification of fumonisins, deoxynivalenol and aflatoxins is described. These assays are field portable, simple to perform, rapid and require no washing steps.

Twenty-five samples of retail corn flakes (from 15 lots) were analysed for fumonisin B(1) (FB(1)) and fumonisin B(2) (FB(2)). They were detected in 22 and 12 samples, respectively, at respective mean concentrations 68 and 8 ng g(-1). Samples were extracted with methanol-acetonitrile-water (25:25:50) and there was an excellent correlation for FB(1) between results obtained with C(18) clean-up and those obtained with the immunoaffinity column (IAC) clean-up. After extraction of the corn flakes' residue with 1% sodium dodecyl sulphate (SDS) solution and hydrolysis with 2 N potassium hydroxide, hidden (protein bound) fumonisin was determined as HFB(1), which was found in residues from all the corn flakes samples, even those containing no detectable FB(1); the average concentration of HFB(1) was 101 ng g(-1), equivalent to 180 ng FB(1) g(-1). Thus, our results showed an average of 2.6 times more FB(1) present in bound form as was determined by conventional analysis. We found a correlation coefficient of -0.5034 for a logarithmic relationship between the FB(1) (C(18) clean-up) and HFB(1) concentrations The highest concentration of HFB(1) formed was 288 ng g(-1) from a sample containing only 12-15 ng FB(1) g(-1), while the lowest concentration of HFB(1) was 26 ng g(-1) from a sample with 152-155 ng FB(1) g(-1). This low degree of correlation should be taken into account by food safety authorities in estimates of human exposure to protein bound fumonisin.