NIR-HSI as a tool to predict deoxynivalenol and fumonisins in maize kernels: a step forward in preventing mycotoxin contamination.

BACKGROUND: Maize is frequently contaminated with deoxynivalenol (DON) and fumonisins B1 (FB1 ) and B2 (FB2 ). In the European Union, these mycotoxins are regulated in maize and maize-derived products. To comply with these regulations, industries require a fast, economic, safe, non-destructive and environmentally friendly analysis method. RESULTS: In the present study, near-infrared hyperspectral imaging (NIR-HSI) was used to develop regression and classification models for DON, FB1 and FB2 in maize kernels. The best regression models presented the following root mean square error of cross validation and ratio of performance to deviation values: 0.848 mg kg-1 and 2.344 (DON), 3.714 mg kg-1 and 2.018 (FB1 ) and 2.104 mg kg-1 and 2.301 (FB2 ). Regarding classification, European Union legal limits for DON and FB1 + FB2 were selected as thresholds to classify maize kernels as acceptable or not. The sensitivity and specificity were 0.778 and 1 for the best DON classification model and 0.607 and 0.938 for the best FB1 + FB2 classification model. CONCLUSION: NIR-HSI can help reduce DON and fumonisins contamination in the maize food and feed chain. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Inbred Selection for Increased Resistance to Kernel Contamination with Fumonisins.

In temperate world-wide regions, maize kernels are often infected with the fumonisin-producing fungus Fusarium verticillioides which poses food and feed threats to animals and humans. As maize breeding has been revealed as one of the main tools with which to reduce kernel contamination with fumonisins, a pedigree selection program for increased resistance to Fusarium ear rot (FER), a trait highly correlated with kernel fumonisin content, was initiated in 2014 with the aim of obtaining inbred lines (named EPFUM) with resistance to kernel contamination with fumonisins and adapted to our environmental conditions. The new released EPFUM inbreds, their parental inbreds, hybrids involving crosses of one or two EPFUM inbreds, as well as commercial hybrids were evaluated in the current study. The objectives were (i) to assess if inbreds released by that breeding program were significantly more resistant than their parental inbreds and (ii) to examine if hybrids derived from EPFUM inbreds could be competitive based on grain yield and resistance to FER and fumonisin contamination. Second-cycle inbreds obtained through this pedigree selection program did not significantly improve the levels of resistance to fumonisin contamination of their parental inbreds; however, most EPFUM hybrids showed significantly better resistance to FER and fumonisin contamination than commercial hybrids did. Although European flint materials seem to be the most promising reservoirs of alleles with favorable additive and/or dominance effects for resistance to kernel contamination with fumonisins, marketable new Reid × Lancaster hybrids have been detected as they combine high resistance and yields comparable to those exhibited by commercial hybrids. Moreover, the white kernel hybrid EPFUM-4 × EP116 exploits the genetic variability within the European flint germplasm and can be an alternative to dent hybrid cultivation because white flint grain can lead to higher market prices.

Quantification and classification of deoxynivalenol-contaminated oat samples by near-infrared hyperspectral imaging.

Deoxynivalenol (DON) is the most occurring mycotoxin in oat and oat-based products. Near-infrared hyperspectral imaging (NIR-HSI) has been proposed as a promising methodology for analysing DON contamination in the food industry. The present study aims to apply NIR-HSI for DON detection in oat kernels and to quantify and classify naturally DON-contaminated oat samples. Unground and ground oat samples were scanned by NIR-HSI before their DON content was determined by HPLC. The data were pre-treated and analysed by PLS regression and four classification methods. The most efficient DON prediction model was for unground samples (R2 = 0.75 and RMSEP = 403.18 µg/kg), using twelve characteristic wavelengths with a special interest in 1203 and 1388 nm. The random forest algorithm of unground samples according to the EU maximum limit for unprocessed oats (1750 µg/kg) achieved a classification accuracy of 77.8 %. These findings indicate that NIR-HSI is a promising tool for detecting DON in oats.

Biocontrol of Mycotoxigenic Fungi Using Bacteria Isolated from Ecological Vineyard Soils.

The presence of mycotoxin-producing Aspergillus species in vineyards is a problem for food safety and the economy. In addition, rising temperatures due to climate change are modifying microbial communities, causing the replacement of some fungal species and the rise of mycotoxins such as aflatoxins. The use of microorganisms as biological control agents (BCAs) is one of the most promising strategies to prevent fungal growth and toxin production. In this study, 513 microorganisms were isolated from organic vineyard soils in different regions of Spain. The 480 bacteria and 33 yeasts isolated were sequentially screened to select those with the most suitable characteristics to be used as BCAs. After identifying 16 isolates meeting all requirements, six bacterial isolates were selected to test their potential to control three relevant toxigenic grape fungi in vitro: A. carbonarius, A. niger and A. flavus. Isolates of Arthrobacter sp., Rhodococcus sp. and Bacillus mycoides showed an excellent ability to reduce the growth and mycotoxin concentration of the above-mentioned fungi and represent potential candidates for further study regarding their possible industrial application as a BCA.

Influence of Agronomic Factors on Mycotoxin Contamination in Maize and Changes during a 10-Day Harvest-Till-Drying Simulation Period: A Different Perspective.

Agronomic factors can affect mycotoxin contamination of maize, one of the most produced cereals. Maize is usually harvested at 18% moisture, but it is not microbiologically stable until it reaches 14% moisture at the drying plants. We studied how three agronomic factors (crop diversification, tillage system and nitrogen fertilization rate) can affect fungal and mycotoxin contamination (deoxynivalenol and fumonisins B1 and B2) in maize at harvest. In addition, changes in maize during a simulated harvest-till-drying period were studied. DON content at harvest was higher for maize under intensive tillage than using direct drilling (2695 and 474 µg kg-1, respectively). We found two reasons for this: (i) soil crusting in intensive tillage plots caused the formation of pools of water that created high air humidity conditions, favouring the development of DON-producing moulds; (ii) the population of Lumbricus terrestris, an earthworm that would indirectly minimize fungal infection and mycotoxin production on maize kernels, is reduced in intensive tillage plots. Therefore, direct drilling is a better approach than intensive tillage for both preventing DON contamination and preserving soil quality. Concerning the simulated harvest-till-drying period, DON significantly increased between storage days 0 and 5. Water activity dropped on the 4th day, below the threshold for DON production (around 0.91). From our perspective, this study constitutes a step forward towards understanding the relationships between agronomic factors and mycotoxin contamination in maize, and towards improving food safety.

Deterministic and Probabilistic Dietary Exposure Assessment to Deoxynivalenol in Spain and the Catalonia Region.

Deoxynivalenol (DON) remains one of the most concerning mycotoxins produced by the Fusarium genus due to the wide occurrence in highly consumed cereal-based food and its associated toxicological effects. Previous studies conducted in Spain and other European countries suggested that some vulnerable groups such as children could be exceeding the tolerable daily intakes. Thus, the aim of this study was to conduct a comprehensive and updated dietary exposure assessment study in Spain, with a specific analysis in the region of Catalonia. Cereal-based food samples collected during 2019 were analysed using liquid chromatography coupled to tandem mass spectrometry for multi-mycotoxin detection including DON and its main metabolites and derivatives. Consumption data were gathered from the nation-wide food surveys ENALIA and ENALIA2 conducted in Spain, and a specific survey conducted in Catalonia. The data were combined using deterministic and semi-parametric probabilistic methods. The results showed that DON was widely present in cereal-based food highly consumed in Spain and the Catalonia region. Exposure to DON among the adult population was globally low; however, among infants aged 3-9 years, it resulted in the median of 192 ng/kg body weight/day and the 95th percentiles of 604 ng/kg body weight/day, that would exceed the most conservative safety threshold for infants. Bread and pasta were the main contributing foodstuffs to the global exposure to DON, even among infants; thus, those foods should be considered a priority for food control or to develop strategies to reduce the exposure. In any case, further toxicological and epidemiological studies are required in order to refine the safety thresholds accounting for the sensitivity of the infant population.

Hyperspectral imaging for the classification of individual cereal kernels according to fungal and mycotoxins contamination: A review.

One of the most common concerns in the cereal industry is the presence of fungi and their associated mycotoxins. Hyperspectral Imaging (HSI) has been proposed recently as one of the most potent tools to manage fungal associated contamination. The introduction of a spatial dimension to the spectral analysis allows the selection of the specific regions of the sample for further screening. Single kernel analysis would enable the discrimination of the highly contaminated kernels to establish a mitigation strategy, overcoming the contamination heterogeneity of cereal batches. This document is a detailed review of the HSI recently published studies that aimed to discriminate fungi and mycotoxin contaminated single cereal kernels. The most relevant findings showed that fungal infection and mycotoxins levels discrimination accuracies were above 90% and 80%, respectively. The results indicate that NIR-HSI is suitable for the detection of fungal-related contamination in single kernels and it has potential to be applied at food industry stages.

Relevant Fusarium Mycotoxins in Malt and Beer.

Mycotoxins are secondary fungal metabolites of high concern in the food and feed industry. Their presence in many cereal-based products has been numerously reported. Beer is the most consumed alcoholic beverage worldwide, and Fusarium mycotoxins originating from the malted and unmalted cereals might reach the final product. This review aims to describe the possible Fusarium fungi that could infect the cereals used in beer production, the transfer of mycotoxins throughout malting and brewing as well as an insight into the incidence of mycotoxins in the craft beer segment of the industry. Studies show that germination is the malting step that can lead to a significant increase in the level of all Fusarium mycotoxins. The first step of mashing (45 °C) has been proved to possess the most significant impact in the transfer of hydrophilic toxins from the grist into the wort. However, during fermentation, a slight reduction of deoxynivalenol, and especially of zearalenone, is achieved. This review also highlights the limited research available on craft beer and the occurrence of mycotoxins in these products.

Occurrence and Dietary Exposure Assessment to Enniatin B through Consumption of Cereal-Based Products in Spain and the Catalonia Region.

Enniatin B (ENNB) is a mycotoxin produced by moulds from the Fusarium genera and its toxic effects are still not fully elucidated, hence a safe reference exposure value has not been established yet. ENNB is the most prevalent emerging mycotoxin and is widely found in cereal-based products, nevertheless, there are no comprehensive exposure assessment studies. For that reason, the aim of this study was to characterise the occurrence of ENNB and estimate the exposure of the Spanish and Catalan populations. A total of 347 cereal-based products were collected in 2019 and were analysed using liquid chromatography-tandem mass spectrometry. Consumption data were obtained from the national food consumption surveys (ENALIA) and a regional survey conducted in Catalonia. The global exposure was estimated using deterministic and probabilistic methods. The results showed a high occurrence of close to 100% in all foodstuffs, with a range from 6 to 269 µg/kg, and a strong correlation with the levels of deoxynivalenol. Children aged one-nine years were the most exposed, showing mean estimates in the range 308-324 ng/kg bw/day and 95th percentiles 697-781 ng/kg bw/day. This study stresses the need for further toxicological data to establish reference doses and conclude formal risk assessment, accounting for the co-occurrence with deoxynivalenol.

Effect of a Mycotoxin Binder (MMDA) on the Growth Performance, Blood and Carcass Characteristics of Broilers Fed Ochratoxin A and T-2 Mycotoxin Contaminated Diets.

The present study was conducted to evaluate the efficacy of the feed additive, a novel multicomponent mycotoxin detoxifying agent (MMDA) containing modified zeolite (clinoptilolite), Bacillus subtilis, B. licheniformis, Saccharomyces cerevisiae cell walls, and silymarin, as detoxifiers of 0.5 mg/kg (0.5 ppm) ochratoxin A (OTA) and 1 mg/kg (1 ppm) T-2 toxin on broiler chickens. A total of 240 1-old broiler chickens (Ross 308) were randomly distributed into five different dietary treatments: (1) control (non-contaminated diet); (2) non contaminated diet + 3 g/kg of MMDA; (3) non-contaminated diet + 0.5 mg/kg OTA + 1 mg/kg T-2 toxin; (4) non-contaminated diet + 0.5 mg/kg OTA + 1 mg/kg T-2 toxin + 1 g/kg MMDA; and (5) non-contaminated diet + 0.5 mg/kg OTA + 1 g/kg T-2 toxin + 3 g/kg MMDA. The results showed that, in the starter period, from 1 to 10 days, the presence of OTA and T-2 mycotoxins reduced the consumption of feed and the growth of the broilers, and no effects of the detoxifying product were observed in the productivity of the chickens, at any of the doses tested, compared to the contaminated control (treatment 3). However, in the growing period, the same negative effect of mycotoxins was registered, but a recovery was observed in the consumption of feed and in the weight of the broilers that consumed 3 g/kg of the MMDA mycotoxin binder, reaching similar values to those of chickens fed uncontaminated control diets. The presence of mycotoxins in feed led to a reduction in the concentration of total proteins and albumin in blood compared to controls, and the presence of the detoxifying product partially reversed this effect. The breast yield of the chickens fed with mycotoxins was lower than that of the animals fed with the control feed and was not affected by the presence of the product tested, at 1 or 3 g/kg. The weight of the different organs (liver, gizzard, kidneys, or spleen), the intestinal pH, the histology of the small intestine, and oral lesions were not affected by the experimental treatments. In summary, the productive parameters and some blood and carcass characteristics of broiler chickens were impaired by the dietary presence of OTA and T-2 toxin. The tested product included at 1 or 3 g/kg feed in contaminated diets improved performance and seems to be effective in partly counteracting the deleterious effects of the tested mycotoxins.

Biomarkers of Deoxynivalenol Toxicity in Chickens with Special Emphasis on Metabolic and Welfare Parameters.

Deoxynivalenol (DON), a trichothecene mycotoxin produced by Fusarium species, is the most widespread mycotoxin in poultry feed worldwide. Long term-exposure from low to moderate DON concentrations can produce alteration in growth performance and impairment of the health status of birds. To evaluate the efficacy of mycotoxin-detoxifying agent alleviating the toxic effects of DON, the most relevant biomarkers of toxicity of DON in chickens should be firstly determined. The specific biomarker of exposure of DON in chickens is DON-3 sulphate found in different biological matrices (plasma and excreta). Regarding the nonspecific biomarkers called also biomarkers of effect, the most relevant ones are the impairment of the productive parameters, the intestinal morphology (reduction of villus height) and the enlargement of the gizzard. Moreover, the biomarkers of effect related to physiology (decrease of blood proteins, triglycerides, hemoglobin, erythrocytes, and lymphocytes and the increase of alanine transaminase (ALT)), immunity (response to common vaccines and release of some proinflammatory cytokines) and welfare status of the birds (such as the increase of Thiobarbituric acid reactive substances (TBARS) and the stress index), has been reported. This review highlights the available information regarding both types of biomarkers of DON toxicity in chickens.

Standardization of near infrared hyperspectral imaging for wheat single kernel sorting according to deoxynivalenol level.

The spatial recognition feature of near infrared hyperspectral imaging (HSI-NIR) makes it potentially suitable for Fusarium and deoxynivalenol (DON) management in single kernels to break with heterogeneity of contamination in wheat batches to move towards individual kernel sorting and provide more quick, environmental-friendly and non-destructive analysis than wet-chemistry techniques. The aim of this study was to standardize HSI-NIR for individual kernel analysis of Fusarium damage and DON presence, to predict the level of contamination and classify grains according to the EU maximum limit (1250 µg/kg). Visual inspection on Fusarium infection symptoms and HPLC analysis for DON determination were used as reference methods. The kernels were scanned in both crease-up and crease-down position and for different image captures. The spectra were pretreated by Multiplicative Scatter Correction (MSC) and Standard Normal Variate (SNV), 1st and 2nd derivatives and normalisation, and they were evaluated also by removing spectral tails. The best fitted predictive model was on SNV pretreated data (R2 0.88 and RMSECV 4.8 mg/kg) in which 7 characteristic wavelengths were used. Linear Discriminant Analysis (LDA), Naïve Bayes and K-nearest Neighbours models classified with 100% of accuracy 1st derivative and SNV pretreated spectra according to symptomatology and with 98.9 and 98.4% of correctness 1st derivative and SNV spectra, respectively. The starting point results are encouraging for future investigations on HSI-NIR technique application to Fusarium and DON management in single wheat kernels to overcome their contamination heterogeneity.

Effects of Deoxynivalenol-Contaminated Diets on Metabolic and Immunological Parameters in Broiler Chickens.

The current study was conducted to examine the effects of deoxynivalenol (DON) at different levels (5 and 15 mg/kg feed) on the metabolism, immune response and welfare parameters of male broiler chickens (Ross 308) at 42 days old. Forty-five 1 day-old broiler chickens were randomly distributed into three different dietary treatments: (1) control, (2) DON-contaminated diet with 5 mg DON/kg of feed (guidance level), and (3) DON-contaminated diet with 15 mg DON/kg of feed. Five replicated cages with three birds each were used for each treatment in a randomized complete block design. The results showed that DON was detected in excreta of birds fed contaminated diets compared with controls. The metabolite DON-3 sulphate (DON-3S) was detected in plasma and excreta in both treated groups, as well as in the liver (but only at 15 mg/kg feed). The increase in the level of DON decreased the hemoglobin concentration (p < 0.001), whereas the erythrocyte counts were only decreased at 15 mg DON/kg feed. No effect of DON on the responses to common vaccines was observed. In plasma, interleukin 8 levels in both contaminated groups were significantly higher than in the control group. The expression of interleukin 6, interleukin 1ß and interferon-γ increased in jejunum tissues of broilers fed 5 mg/kg of DON compared with controls. The stress index (heterophil to lymphocyte ratio) was not affected by DON-contaminated diets compared with controls. The plasma corticosterone level was significantly lower in both DON groups compared with controls. In conclusion, DON-3S could be used as a specific biomarker of DON in different biological matrices, while the immune response in broiler chickens is stimulated by the presence of DON at the guidance level, but no adverse effect was observed on physiological stress parameters.

Near-infrared hyperspectral imaging for deoxynivalenol and ergosterol estimation in wheat samples.

The present study aimed to evaluate the use of hyperspectral imaging (HSI)-NIR spectroscopy to assess the presence of DON and ergosterol in wheat samples through prediction and classification models. To achieve these objectives, a first set of bulk samples was scanned by HSI-NIR and divided into two subsamples, one that was analysed for ergosterol and another that was analysed for DON by HPLC. This method was repeated for a second larger set to build prediction and classification models. All the spectra were pretreated and statistically processed by PLS and LDA. The prediction models presented a RMSEP of 1.17 mg/kg and 501 µg/kg for ergosterol and DON, respectively. Classification achieved an encouraging accuracy of 85.4% for an independent validation set of samples. The results confirm that HSI-NIR may be a suitable technique for ergosterol quantification and DON classification of samples according to the EU legal limit for DON.

Effects of Deoxynivalenol-Contaminated Diets on Productive, Morphological, and Physiological Indicators in Broiler Chickens.

The present study with 1-day-old male broilers (Ross 308) was conducted to evaluate the effects of deoxynivalenol (DON) at different levels (5 and 15 mg/kg feed) on growth performance, relative weight of organs, morphology of the small intestine, serum biochemistry, and welfare parameters of broiler chickens. Forty-five broiler chicks were randomly divided into three different experimental groups with five replicates each: (1) control group received a non-contaminated diet, (2) contaminated diet with 5 mg DON/kg of feed, and (3) contaminated diet with 15 mg DON/kg of feed for 42 days. Results showed that feed artificially contaminated with DON at guidance level (5 mg/kg diet) did not affect growth performance parameters. However, 15 mg/kg reduced body weight gain and altered feed efficiency. DON at two assayed levels significantly increased the absolute and relative weight of thymus and the relative weight of gizzard and decreased the absolute and the relative weight of the colon. Compared to controls, both doses affected small intestine morphometry parameters. In terms of biochemical indicators, DON at 5 mg/kg reduced the creatine kinase level and at 15 mg/kg DON reduced the cholesterol level. Furthermore, DON at 15 mg/kg induced more fear in broilers compared to broilers fed the guidance level. It was concluded that even the guidance level of DON did not affect the chickens' performance. However, its toxic effect occurred in some organs and biochemical parameters.

Tri-octahedral bentonites as potential technological feed additive for Fusarium mycotoxin reduction.

In 2009 the EU Regulation 386/2009 established a new functional group of feed additives called "substances for reduction of the contamination of feed by mycotoxin". Later, di-octahedral bentonite (1 m558) was authorised, as an anti-aflatoxin additive, being the only additive of this group authorised to date. This work aims to demonstrate the effectiveness of other bentonites, such as tri-octahedral bentonites, versus Fusarium-mycotoxins, since very few adsorbents have proved their effectiveness in relation to this group of mycotoxins. For this purpose, 7 bentonites (six of them tri-octahedral) and 7 commercial adsorbents, added at 0.02% (w/v), were assayed in an in vitro adsorption experiment using two simulated gastro-intestinal (GI) juices (pH 1.3 and 6.8) versus zearalenone (ZEN: 0.1-5 mg/L), fumonisin B1 (FB1: 1-10 mg/L) and deoxynivalenol (DON: 2-10 mg/L). Mycotoxin adsorption data were fitted to Langmuir and Freundlich isotherms. In vitro adsorption experiments showed that ZEN and FB1 (in the latter case only in acid medium) were partially adsorbed, while the adsorption of DON was negligible. Moreover, the increase of adsorbent dose (up to 0.20%, w/v) significantly improved the in vitro adsorption of ZEN and FB1, reaching >90% of adsorption. The present work proposes the use of some tri-octahedral bentonites as feed additives for Fusarium-mycotoxin reduction.

Fusarium mycotoxins in total mixed rations for dairy cows.

Mycotoxins produced by certain fungal species of the Fusarium genus are frequently found as contaminants in cereals and feedstuffs. Fumonisins (FBs), deoxynivalenol (DON) and zearalenone (ZEN) are of special concern relative to animal health and productivity. The aim of this work was to analyse the levels of Fusarium mycotoxin contamination in samples of total mixed rations (TMRs) for dairy cows. To accomplish this analysis, an HPLC-MS/MS multi-mycotoxin method was developed and validated. The relation between the formulation of TMR samples and the presence of mycotoxins was also studied. From February 2016 to January 2018, a total of 193 TMR samples for dairy cows collected from farms located in different areas of Spain were analysed for the presence of FBs, ZEN, DON and their metabolites. In total, 112 samples (58%) were contaminated with at least one mycotoxin, and 38 samples (20%) presented more than one mycotoxin. FBs were the mycotoxins most frequently found (34% positive samples). DON was detected in 17% of samples, and ZEN was detected in 16% of samples. Among the metabolites analysed, only deoxynivalenol-3-glucoside (DON-3-Glc) and 15-acetyldeoxynivalenol (15-ADON) were detected. The levels of all the Fusarium mycotoxins studied were always below the values recommended by the European Commission for feedstuffs. The wide variety of ingredients used in the formulation of the analysed samples made it difficult to reach definite conclusions, although it seemed that some cereal silages and concentrates such as cereals or compound feed used as ingredients of the TMR may be related to the presence of mycotoxins.

The fate of Fusarium mycotoxins (deoxynivalenol and zearalenone) through wort fermenting by Saccharomyces yeasts (S. cerevisiae and S. pastorianus).

The aim of this study was to evaluate the effect of 15 commercial yeasts in the mitigation of the Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) during the brewing process. Saccharomyces strains (10 strains of S. cerevisiae and 5 of S. pastorianus) were used to ferment DON and ZEN contaminated wort. Wort samples were taken every 24 h during fermentation, while mycotoxin analysis in yeast was performed at the end of fermentation (96 h); additionally, pH and ethanol content were measured daily. For mycotoxin analysis, after immunoaffinity purification of sample extracts, analysis was performed using an Ultra-High-Pressure Liquid Chromatograph coupled with a diode array or fluorescence detector (UHPLC-DAD/FLD). Mycotoxin presence had no significant effect on the ethanol production during brewing. At the end of fermentation, 10-17% of DON and 30-70% of ZEN had been removed, 6% of the initial concentration of DON and 31% of the ZEN being adsorbed by the yeast. Beermakers must pay careful attention to the raw material since a high percentage of DON could be present at the end of the beer fermentation process. Future studies should focus on the quantification of "masked" mycotoxins that are relevant to food security.

Frequency and levels of mycotoxins in beer from the Mexican market and exposure estimate for deoxynivalenol mycotoxins.

The aim of the present study was to evaluate the occurrence of 23 mycotoxins in beer purchased in Mexico and to assess two exposure scenarios in the Mexican population through beer consumption. Multi-mycotoxin analysis of a total of 61 different beers (132 samples) was carried out using UHPLC-MS/MS equipment. Probability density functions were used to describe mycotoxins contamination. The daily intake of mycotoxins was estimated using a semi-probabilistic approach, applying the Monte Carlo method. Deoxynivalenol (DON) and its metabolites (deoxynivalenol-3-glucoside (DON3G) and 3-acetyl-deoxynivalenol (3ADON)) were the mycotoxins found in higher proportions in contaminated samples. None of the other mycotoxins overpassed the limit of quantification (LOQ) of the method. The combined intake of DON and its analogues ranged from 5.24 to 86.59 ng kg-1 bw day-1, which represent from 1.20 to 19.83% of the DON TDI. The results suggest that depending on the individual consumption of beer and depending on the type of beer, the intake of DON via beer could represent a significant percentage of the tolerable daily intake (TDI).

Transfer of Fusarium mycotoxins from malt to boiled wort.

The fate of deoxynivalenol, deoxynivalenol-3-glucoside, 3- and 15-acetyl-deoxynivalenol, zearalenone, α- and ß-zearalenol and fumonisins (fumonisin B1 and fumonisin B2) through mashing and wort boiling was studied. Three different mycotoxin contamination scenarios were considered. In almost all samples an increase in the level of mycotoxins in wort was observed during mashing followed by a decrease after just 30 min of the process, with levels remaining constant until the end of boiling. Deoxynivalenol and its metabolites were reduced to their initial level contained in the malt before mashing, or even lower, however in none of the samples they were completely eliminated. Zearalenone was not quantitated at the end of boiling, although there was a significant initial level of ZEN. ß-Zearalenol remained unaltered during the process. Fumonisins were reduced by between 50 and 100 per cent during mashing and boiling.