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The handling of data on food contamination frequently represents a challenge because these are often left-censored, being composed of both positive and non-detected values. The latter observations are not quantified and provide only the information that they are below a laboratory-specific threshold value. Besides deterministic approaches, which simplify the treatment through the substitution of non-detected values with fixed threshold or null values, a growing interest has been shown in the application of stochastic approaches to the treatment of unquantified values. In this study, a multiple imputation procedure was applied in order to analyze contamination data on deoxynivalenol, a mycotoxin that may be present in pasta and pasta substitute products. An application of the proposed technique to censored deoxynivalenol occurrence data is presented. The results were compared to those attained using deterministic techniques (substitution methods). In this context, the stochastic approach seemed to provide a more accurate, unbiased and realistic solution to the problem of left-censored occurrence data. The complete sample of values could then be used to estimate the exposure of the general population to deoxynivalenol based on consumption data.

This study presents the exposure scenario to mycotoxins of adult population throughout Europe. The urinary biomarkers values were obtained by modelling data from two European projects. Exposure to AFB1, OTA, CIT, FBs, DON, NIV and T2/HT2 are presented. The main output obtained refers to a concern for public health about AFM1, FBs, T2/HT2 and NIV, and low concern for OTA, DON and CIT. The margin of exposure for AFM1 did not respect the reference value of 10,000 considered of low priority for risk; for Fusarium toxins, FBs and T2/HT2, probable daily intake (PDI) values resulted about ten times higher than their tolerable daily intake and NIV presented the most critical situation with a calculated PDI 30 times higher than the reference TDI value. North and South Europe scenarios were also depicted by clustering biomonitoring data. OTA and DON showed to be prevalent in Northern countries and the opposite was noticed for ZEN, higher in Southern countries. The critical issues of the availability of records feeding the dataset and of the accuracy of excretion rate for some mycotoxins are source of uncertainty for the reliability of the outputs, nevertheless the time is ripe for asking for more concrete HBM values and/or HBM-HBGV which would help in interpreting the burden of mycotoxins in Europe.

Exposure to environmental contaminants might play an important role in neurodegenerative disease pathogenesis, such as Parkinson´s disease (PD) and Alzheimer´s disease (AD). For the first time in Spain, the plasmatic levels of 19 mycotoxins from patients diagnosed with a neurodegenerative disease (44 PD and 24 AD) and from their healthy companions (25) from La Rioja region were analyzed. The studied mycotoxins were aflatoxins B1, B2, G1, G2 and M1, T-2 and HT-2, ochratoxins A (OTA) and B (OTB), zearalenone, sterigmatocystin (STER), nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deepoxy-deoxynivalenol, neosolaniol, diacetoxyscirpenol and fusarenon-X. Samples were analyzed by LC-MS/MS before and after treatment with β-glucuronidase/arylsulfatase in order to detect potential metabolites. Only OTA, OTB and STER were detected in the samples. OTA was present before (77% of the samples) and after (89%) the enzymatic treatment, while OTB was only detectable before (13%). Statistically significant differences in OTA between healthy companions and patients were observed but the observed differences might seem more related to gender (OTA levels higher in men, p -value = 0.0014) than the disease itself. STER appeared only after enzymatic treatment (88%). Statistical analysis on STER, showed distributions always different between healthy controls and patients (patients' group > controls, p -value < 0.0001). Surprisingly, STER levels weakly correlated positively with age in women (rho = 0.3384), while OTA correlation showed a decrease of levels with age especially in the men with PD (rho = -0.4643).

Introduction: Turmeric is the common name for the rhizome of Curcuma longa L. In the recent years, food supplements containing turmeric have been marketed and widely used by an increasing number of consumers. Spontaneous reports of suspected adverse reactions to food supplements are collected within the Phytovigilance system., Methods: An ad hoc multidisciplinary group investigated the suspected cases of hepatotoxicity reported to the Italian Phytovigilance system associated with the assumption of turmeric food supplements with the methodology specific to pharmacovigilance as well as for the evaluation of the quality and safety of food supplements., Results: A cluster of 28 spontaneous reports of acute hepatitis, mostly with cholestasis, associated with turmeric products were sent to the Italian Phytovigilance system in the first six months of 2019. In all cases, except one, the causality assessment was at least possible. The suspected products were collected and analysed for the presence of drugs, heavy metals, aflatoxins, pesticides, synthetic dyes and pyrrolizidine alkaloids., Conclusion: On the basis of the results of all the activities performed by multidisciplinary group, regulatory intervention was taken. This study highlights the importance of developing an integrated evaluation approach for the evaluation of the adverse effects associated with the use of food supplements.

The aim of this analysis was to assess the association between consumption of maize and dairy products and urine and serum levels of aflatoxin FM1 (AFM1) in a sample of 59 males occupationally exposed (29) and non-exposed (30) to aflatoxins. Two urine samples were collected for each person; each sample was accompanied by a questionnaire on food consumption in the preceding 96 h. Given the similar levels of contamination found in exposed and non-exposed workers, the association between food consumption and AFM1 levels was analyzed by pooling samples from exposed and non-exposed workers. No serum sample was found to be positive for AFM1, whereas 74% of the urine samples were positive; the average concentration of positive samples was 0.042 ng/mL (range < limit of detection (LoD) (0.002)-0.399 ng/mL). Of the 21 samples from maize consumers, 13 were positive for AFM1 (62%), with a mean concentration of 0.026 ng/mL (range 0.006-0.088 ng/mL), while 76% (74/94) of the samples from maize non-consumers were positive (mean 0.045, range < LoD (0.002)-0.399 ng/mL). No association was found with milk or dairy products. The high urine level of aflatoxins found in both exposed and non-exposed workers was not associated with the consumption of maize or cow milk products.

Deoxynivalenol (DON) is a mycotoxin mainly produced by Fusarium graminearum that can contaminate cereals and cereal-based foodstuff. Urinary DON levels can be used as biomarker for exposure assessment purposes. This study assessed urinary DON concentrations in Italian volunteers recruited by age group, namely children, adolescents, adults, and the elderly. In addition, vulnerable groups, namely vegetarians and pregnant women, were included in the study. To determine the urinary DON, its glucuronide and de-epoxydated (DOM-1) forms, an indirect analytical approach was used, measuring free DON and total DON (as sum of free and glucuronides forms), before and after enzymatic treatment, respectively. Morning urine samples were collected on two consecutive days, from six different population groups, namely children, adolescent, adults, elderly, vegetarians and pregnant women. Total DON was measured in the 76% of the collected samples with the maximum incidences in children and adolescent age group. Urine samples from children and adolescent also showed the highest total DON levels, up to 17.0 ng/mg creat . Pregnant women had the lowest positive samples per category (40% for day 1 and 43% for day 2, respectively), low mean levels of total DON (down to 2.84 ng/mg creat ) and median equal to 0 ng/mg creat . Estimation of DON dietary intake reveals that 7.5% of the total population exceeds the TDI of 1 μg/kg bw/day set for DON, with children showing 40% of individuals surpassing this value (male, day 2)., Competing Interests: The authors declare no conflict of interest.

Mycotoxins exposure by inhalation and/or dermal contact is possible in different branches of industry especially where heavily dusty settings are present and the handling of dusty commodities is performed. This study aims to explore the validity of the biomonitoring as a tool to investigate the intake of mycotoxins in a population of workers operating in an Italian feed plant. Serum samples were collected for the determination of aflatoxins B1 (AFB1), AFB1-Lysine adduct and ochratoxin A (OTA). A method based on liquid-liquid extraction coupled with high resolution mass spectrometry determination was developed and fully validated. For AFB1, a high number of non-detected samples (90%) was found and no statistical difference was observed comparing workers and control group. None of the analyzed samples showed the presence of AFB1-Lysine adduct. For OTA, the 100% of the analyzed samples was positive with a 33% of the samples showing a concentration higher than the limit of quantification (LOQ), but no statistical difference was highlighted between the average levels of exposed and control groups. In conclusion, the presence of AFB1 and OTA in serum cannot be attributable to occupational exposure.

Genus Claviceps is a plant pathogen able to produce a group of toxins, ergot alkaloids (EAs), whose effects have been known since the Middle Ages (ergotism). Claviceps purpurea is the most important representative specie, known to infect more than 400 monocotyledonous plants including economically important cereal grains (e.g., rye, wheat, triticale). EAs are not regulated as such. Maximum limits are in the pipeline of the EU Commission while at present ergot sclerotia content is set by the Regulation (EC) No. 1881/2006 in unprocessed cereals (0.05% as a maximum). This study aimed to investigate the presence of the six principal EAs (ergometrine, ergosine, ergocornine, α-ergocryptine, ergotamine and ergocristine) and their relative epimers (-inine forms) in rye- and wheat-based products. Of the samples, 85% resulted positive for at least one of the EAs. Wheat bread was the product with the highest number of positivity (56%), followed by wheat flour (26%). Rye and wheat bread samples showed the highest values when the sum of the EAs was considered, and durum wheat bread was the more contaminated sample (1142.6 μg/kg). These results suggest that ongoing monitoring of EAs in food products is critical until maximum limits are set.

Deoxynivalenol (DON), the mycotoxin produced mainly by Fusarium graminearum and found in contaminated cereal-based foodstuff, has been consistently detected in body fluids in adults. Available data in children and adolescents are scarce. This study assessed urinary DON concentrations in children aged 3-9 years ( n = 40) and adolescents aged 10-17 years ( n = 39) in the UK. Morning urine samples were collected over two consecutive days and analysed for free DON (un-metabolised form), DON-glucuronides (DON-GlcA), deepoxy deoxynivalenol (DOM-1), and total DON (sum of free DON, DON-GlcA, and DOM-1). Total DON was detected in the urine of >95% of children and adolescents on both days. Mean total DON concentrations (ng/mg creatinine) were 41.6 and 21.0 for children and adolescents, respectively. The greatest total DON levels were obtained in female children on both days (214 and 219 ng/mg creatinine on days 1 and 2, respectively). Free DON and DON-GlcA were detected in most urine specimens, whereas DOM-1 was not present in any sample. Estimation of dietary DON exposure suggested that 33-63% of children and 5-46% of adolescents exceeded current guidance regarding the maximum provisional tolerable daily intake (PMTDI) for DON. Although moderate mean urinary DON concentrations were shown, the high detection frequency of urinary DON, the maximum biomarker concentrations, and estimated dietary DON exposure are concerning., Competing Interests: The authors declare no conflict of interest. EFSA disclaimer: EFSA is not responsible for the views displayed in the publications and/or in conjunction with the activities for which the grant is used. The funding sponsors had a role in the design of the study; Clear information regarding the required design of the study was given in EFSAs call for proposal. Further details can be found at: .

Environmental factors and genetic susceptibility are implicated in the increased risk of autism spectrum disorder (ASD). Mycotoxins are agricultural contaminants of fungal origin that represent real risk factors for human health and especially for children. Thus, the main hypothesis of this work is that the deterioration of the clinical manifestation of autism in children may result from the exposure to mycotoxins through the consumption of contaminated food. Within a cross-sectional study, a group of autistic children ( n = 172) and a group of controls ( n = 61) (siblings and non-parental) were recruited in North and South Italy. All children had blood and urine samples taken, for testing some mycotoxins by a LC-MS/MS validated method. Blood samples were also tested for assessing specific IgG against food and fungal antigens and cytokines. The analyses outputs highlighted statistically significant differences comparing mycotoxins levels between (i) children groups both in urine (deoxynivalenol and de-epoxydeoxynivalenol, p = 0.0141 and p = 0.0259, respectively) and serum (aflatoxin M1, ochratoxin A and fumonisin B1, p = 0.0072, p = 0.0141 and p = 0.0061, respectively); (ii) a group of selected fungal IgGs, and IgGs against wheat and gluten and (iii) cytokines. These results suggest the need for a deeper examination of the role that mycotoxins may have on the etiology of ASD., Competing Interests: The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the result.

Deoxynivalenol (DON) is produced by Fusarium graminearum and is one of the most commonly occurring trichothecenes. Vegetarians are alleged to be a high-risk group for DON exposure due to high intakes of cereals susceptible to the growth of the mycotoxin. This study provides the levels of DON and de-epoxi Deoxynivalenol (DOM-1) in urine analysed by liquid chromatography-mass spectrometry (LC-MS) in UK vegetarians. Over two consecutive days, morning urine samples were collected from 32 vegetarians and 31 UK adult volunteers, and associated food consumption 24 h prior to the sample was recorded. Statistically significant differences between the weight of the UK adults and vegetarians ( t = 3.15. df = 61, p ≤ 0.005 two-tailed) were observed. The mean levels of DON in urine for adults on day 1 was 3.05 ng free DON/mg creatinine, and on day 2 was 2.98 ng free DON/mg creatinine. Even though high mean levels were observed, most adults were within the tolerable daily intake. However, for vegetarians, the mean level of urinary DON on day 1 was 6.69 ng free DON/mg creatinine, and on day 2 was 3.42 ng free DON/mg creatinine. These levels equate to up to 32% of vegetarians exceeding recommended tolerable daily intakes (TDI) of exposure (1 µg/kg b.w./day)., Competing Interests: The authors declare no conflict of interest. EFSA disclaimer: EFSA is not responsible for the views displayed in the publications and/or in conjunction with the activities for which the grant is used. The funding sponsors had a role in the design of the study; Clear information regarding the required design of the study was given in EFSA’s call for proposal. Further details can be found at .

The analytical scenario for determining contaminants in the food and feed sector is constantly prompted by the progress and improvement of knowledge and expertise of researchers and by the technical innovation of the instrumentation available. Mycotoxins are agricultural contaminants of fungal origin occurring at all latitudes worldwide and being characterized by acute and chronic effects on human health and animal wellness, depending on the species sensitivity. The major mycotoxins of food concern are aflatoxin B1 and ochratoxin A, the first for its toxicity, and the second for its recurrent occurrence. However, the European legislation sets maximum limits for mycotoxins, such as aflatoxin B1, ochratoxin A, deoxynivalenol, fumonisins, and zearalenone, and indicative limits for T-2 and HT-2 toxins. Due to the actual probability that co-occurring mycotoxins are present in a food or feed product, nowadays, the availability of reliable, sensitive, and versatile multi-mycotoxin methods is assuming a relevant importance. Due to the wide range of matrices susceptible to mycotoxin contamination and the possible co-occurrence, a multi-mycotoxin and multi-matrix method was validated in liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the purpose to overcome specific matrix effects and analyze complex cereal-based samples within the Italian Total Diet Study project., Competing Interests: The authors declare no conflict of interest.

Feed mill workers may handle or process maize contaminated with aflatoxins (AFs). This condition may lead to an unacceptable intake of toxins deriving from occupational exposure. This study assessed the serological and urinary levels of AFs in workers exposed to potentially contaminated dusts in two mills. From March to April 2014, blood and urine samples were collected, on Monday and Friday morning of the same working week from 29 exposed workers and 30 non-exposed controls. AFs (M₁, G₂, G₁, B₁, B₂) and aflatoxicol (AFOH) A were analyzed. Each subject filled in a questionnaire to evaluate potential food-borne exposures to mycotoxins. AFs contamination in environmental dust was measured in both plants. No serum sample was found to be positive. Seventy four percent of urine samples (73.7%) revealed AFM₁ presence. AFM₁ mean concentration was 0.035 and 0.027 ng/mL in exposed and non-exposed workers, respectively ( p = 0.432); the concentration was slightly higher in Friday's than in Monday's samples, in exposed workers, 0.040 versus (vs.) 0.031 and non-exposed controls (0.030 vs. 0.024, p = 0.437). Environmental AFs contamination ranged from 7.2 to 125.4 µg/kg. The findings of this study reveal the presence of higher AFs concentration in exposed workers than in non-exposed controls, although these differences are to be considered consistent with random fluctuations.

Deoxynivalenol (DON) is one of the most commonly occurring trichothecenes, produced mainly by Fusarium graminearum . Little is known about the effect of DON exposure or the levels of DON exposure that occur during pregnancy. The project aimed to provide data on levels of total DON and de-epoxi Deoxynivalenol (DOM-1) in pregnant human urine samples analysed by liquid chromatography-mass spectrometry (LC-MS). Morning urine samples were collected over two consecutive days from 42 volunteers and associated food consumption was recorded for the 24 h prior to the sample. Spearman's rho non-parametric test for correlation was used to assess the data. Levels of DON did not differ significantly between day 1 (mean 29.7 ng/mL urine or 40.1 ng DON/mg creatinine) and day 2 (mean 28.7 ng/mL urine or 38.8 ng DON/mg creatinine ng/mL/day) urine samples. The only significant positive correlation was found between total ng DON/mg creatinine and parity (rho = 0.307, n = 42, p < 0.005 two-tailed) and total ng DON/mg creatinine with baked goods on day 1 (rho = 0.532, n = 42, p < 0.0005 two-tailed). This study provides data on the DON levels in pregnancy in this suburban population and reassurance that those levels are within acceptable limits., Competing Interests: The authors declare no conflict of interest. EFSA disclaimer: EFSA is not responsible for the views displayed in the publications and/or in conjunction with the activities for which the grant is used. The funding sponsors had a role in the design of the study; Clear information regarding the required design of the study was given in EFSAs call for proposal. Further details can be found at http://www.efsa.europa.eu/en/art36grants/article36/gpefsacontam201304.

Four hundred and seventy-two pasta samples were collected from long retail distribution chain sales points located in North, Central and South Italy. Representative criteria in the sample collection were followed in terms of number of samples collected, market share, and types of pasta. Samples were analysed by an accredited HPLC-UV method of analysis. The mean contamination level (64.8 μg/kg) of deoxynivalenol (DON) was  in the 95th percentile (239 μg/kg) and 99th percentile (337 μg/kg), far below the legal limit (750 μg/kg) set by Regulation EC/1126/2007, accounting for about one tenth, one third and half the legal limit, respectively. Ninety-nine percent of samples fell below half the legal limit. On the basis of the obtained occurrence levels and considering the consumption rates reported by the Italian official database, no health concern was assessed for all consumer groups, being that exposure was far below the Tolerable Daily Intake (TDI) of 1000 ng/kg b.w/day. Nevertheless, despite this, particular attention should be devoted to the exposure to DON by high consumers, such as children aged 3-5 years, who could reach the TDI even with very low levels of DON contamination.

In this paper, we present an approach to format national food consumption data at raw agricultural commodity (RAC) level. In this way, the data is both formatted in a harmonised way given the comparability of RACs between countries, and suitable to assess the dietary exposure to chemicals analysed in RACs at a European level. In this approach, consumption data needs to be converted to edible part of RAC (e-RAC) level using a RAC conversion database. To subsequently use this data in exposure assessments, both e-RACs and RACs analysed in chemical control programmes should be classified via a uniform system. Furthermore, chemical concentrations in RACs may need to be converted to e-RAC level using processing factors. To illustrate the use of this approach, we describe how the Dutch RAC conversion database was used to convert consumption data of four national consumption surveys to e-RAC level, and the use of the FAO/WHO Codex Classification system of Foods and Animal Feeds to harmonise the classification. We demonstrate that this approach works well for pesticides and glycoalkaloids, and is an essential step forward in the harmonisation of risk assessment procedures within Europe when addressing chemicals analysed in RACs by all national food control systems.

Probabilistic dietary acute exposure assessments of captan and tolylfluanid were performed for the populations of the Czech Republic, Denmark, Italy, the Netherlands and Sweden. The basis for these assessments was national databases for food consumption and pesticide concentration data harmonised at the level of raw agricultural commodity. Data were obtained from national food consumption surveys and national monitoring programmes and organised in an electronic platform of databases connected to probabilistic software. The exposure assessments were conducted by linking national food consumption data either (1) to national pesticide concentration data or (2) to a pooled database containing all national pesticide concentration data. We show that with this tool national exposure assessments can be performed in a harmonised way and that pesticide concentrations of other countries can be linked to national food consumption surveys. In this way it is possible to exchange or merge concentration data between countries in situations of data scarcity. This electronic platform in connection with probabilistic software can be seen as a prototype of a data warehouse, including a harmonised approach for dietary exposure modelling.

The aim of this study was to investigate the distribution of fumonisin B1 in various corn milling fractions processed by an industrial plant. Corn kernels and six derived milling fractions (germ, bran, large and small grits, animal feed flour, and flour) were sampled. In addition, in order to evaluate the effect of cooking, samples of polenta were prepared starting from naturally contaminated flour obtained from the industrial processing cycle. The industrial plant worked continuously at a rate of 60 tons per day. Two sublots of 5 tons each were investigated with samples of derived products taken at regular time intervals. Due to a similar heterogeneous distribution of fumonisin B1 with other mycotoxins, such as aflatoxins, the sampling scheme was derived from the European Directive 98/53 for aflatoxins. Both lots of kernels showed fumonisin contamination at 4.54 and 5.09 mg/kg, respectively. Germ, bran, and animal feed flour showed contamination levels, namely 8.92 mg/kg (lot 1) and 9.56 mg/kg (lot 2), 7.08 mg/kg (lot 1) and 8.08 mg/kg (lot 2), and 9.36 mg/kg (lot 1) and 6.86 mg/kg (lot 2) higher than large and small grits and flour (0.39 mg/kg [lot 1] and 0.42 mg/kg [lot 2], 0.60 mg/kg [lot 1] and 1.01 mg/kg [lot 2], and 0.40 mg/kg [lot 1] and 0.45 mg/kg [lot 2], respectively). These results seem to account both for the industrial yields of the derived products and the distribution of fumonisin contamination in a kernel. The cooking of polenta in a domestic pressure cooker did not affect fumonisin contamination because the mycotoxin concentrations were similar to those of the starting flour (0.40 and 0.45 mg/kg).