Evaluation of Statistical Treatment of Left-Censored Contamination Data: Example Involving Deoxynivalenol Occurrence in Pasta and Pasta Substitute Products.

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.

Overall Exposure of European Adult Population to Mycotoxins by Statistically Modelled Biomonitoring 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.

Biomonitoring of Mycotoxins in Plasma of Patients with Alzheimer's and Parkinson's Disease.

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 ß-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).

Turmeric (Curcuma longa L.) food supplements and hepatotoxicity: an integrated evaluation approach.

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.

Association between Urinary Levels of Aflatoxin and Consumption of Food Linked to Maize or Cow Milk or Dairy Products.

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.

Optimization and validation of a LC-HRMS method for aflatoxins determination in urine samples.

Mycotoxins' exposure by inhalation and/or dermal contact can occur 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 possible contribution of the occupational exposure to aflatoxins by analysing urine samples for the presence of aflatoxins B1 and M1 and aflatoxin B1-N7-guanine adduct. The study was conducted in 2017 on two groups of volunteers, the workers group, composed by personnel employed in an Italian feed plant (n = 32), and a control group (n = 29), composed by the administrative employees of the same feed plant; a total of 120 urine samples were collected and analysed. A screening method and a quantitative method with high-resolution mass spectrometry determination were developed and fully validated. Limits of detections were 0.8 and 1.5 pg/mLurine for aflatoxin B1 and M1, respectively. No quantitative determination was possible for the adduct aflatoxin B1-N7-guanine. Aflatoxin B1 and its adduct were not detected in the analysed samples, and aflatoxin M1, instead, was found in 14 samples (12%) within the range 1.9-10.5 pg/mLurine. Only one sample showed a value above the limit of quantification (10.5 pg/mLurine). The absence of a statistical difference between the mean values for workers and the control group which were compared suggests that in this specific setting, no professional exposure occurs. Furthermore, considering the very low level of aflatoxin M1 in the collected urine samples, the contribution from the diet to the overall exposure is to be considered negligible.

Determination of ochratoxin A in pork meat products: single laboratory validation method and preparation of homogeneous batch materials.

Ochratoxin A is one of the most diffused mycotoxin present in a large spectrum of food commodities, mainly produced by Aspergillus ochraceus, Aspergillus carbonarius, Aspergillus niger and Penicillium verrucosum. EU has set maximum limits for a number of matrices such as cereals, wine, spices and liquorice, whilst other commodities such as beer and meat products that are susceptible of OTA contamination and are largely consumed are not included. In 2013, within the framework of the Regulation (EC) 882/2004 on official controls, the European Commission issued the mandate M/520 regarding the standardisation for methods of analysis for mycotoxins in food to the European Committee for Standardisation. Of the 11 priorities of the mandate, the one on "HPLC determination of OTA in meat, meat products and edible offal" was assigned to the Italian National Reference Laboratory for feed and food. The method was single-laboratory validated, and all the performance characteristics of the method were compliant with the corresponding reference values indicated in Regulation (EC) n. 401/2006. The method was applied to characterise a set of 5 pork-based materials (ham, kidney, liver and canned chopped pork) to be used for an inter-laboratory method validation study. Three ham materials (levels of contamination of 0.77, 2.22 and 12.3 µg/kg, respectively), one liver material (contamination level of 2.80 µg/kg) and one chopped pork meat (contamination level of 0.66 µg/kg) were tested for homogeneity and stability.

Determination of Deoxynivalenol Biomarkers in Italian Urine Samples.

Deoxynivalenol (DON) is a mycotoxin mainly produced by Fusariumgraminearum 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/mgcreat. 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/mgcreat) and median equal to 0 ng/mgcreat. 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).

Biomonitoring Data for Assessing Aflatoxins and Ochratoxin A Exposure by Italian Feedstuffs Workers.

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.

Ergot Alkaloids in Wheat and Rye Derived Products in Italy.

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.

Role of mycotoxins in the pathobiology of autism: A first evidence.

Objectives: Gene-environment interaction is an emerging hypothesis to expound not only the autism pathogenesis but also the increased incidence of neurodevelopmental disorders (such as autistic spectrum disorder, attention-deficit, hyperactivity disorder). Among xenobiotics, mycotoxins are worldwide contaminants of food that provoke toxicological effects, crucially resembling several symptoms associated with autism such as oxidative stress, intestinal permeability, and inflammation. Here, we focused on a group of mycotoxins to test their role in the manifestation of autism, try to explain their mechanism of action, and discuss possible preventive and therapeutic interventions. Methods: Autistic children (n = 52) and healthy children [n = 58 (31 siblings and 27 unrelated subjects)] were recruited and body fluids and clinical data collected. The diagnosis of autism was made according to DSM V criteria, then with GMDS 0-2, WPPSI, and ADOS. Ochratoxin A (OTA), gliotoxin, zearalenone, and sphingosine/sphinganine ratio were determined by LC analysis in sera and urines. Statistical analysis was performed by the Wilcoxon Rank Sum (Mann-Whitney) test and Spearman test. Results: By comparing the results of autistic patients with those of unrelated controls, a significant association was found for OTA levels in urines (P = 0.0002) and sera (P = 0.0017), and also comparing patients with siblings and unrelated controls together (P = 0.0081). Discussion: Our results are the first describing a possible role of OTA in the pathobiology of autism. Recalling the male prevalence of ASD (male/female = 4-5/1), it is noted that, in animal models, OTA exerts its neurotoxicity especially in males. Moreover, in vitro, OTA increases microRNA-132 that is dysregulated in autistic patients and involved in reciprocal regulation of the autism-related genes MeCP2 and PTEN. A personalized diet coupled with probiotic administration, especially OTA adsorbing Lactobacillus, could ameliorate autistic symptoms in OTA-positive patients.

Occurrence of deoxynivalenol in an elderly cohort in the UK: a biomonitoring approach.

Deoxynivalenol (DON) is a Fusarium toxin, to which humans are frequently exposed via diet. Although the elderly are speculated to be sensitive to the toxic effects of DON as a result of age-related conditions, disease and altered DON metabolism, there is lack of available data on DON biomarkers in this age group. This study characterised urinary DON concentrations and its metabolites in elderly aged ≥65years (n = 20) residing in Hull, UK. Morning urinary specimens were collected over two consecutive days together with food records to assess dietary intake over a 24h-period prior to each urinary collection. Free DON (un-metabolised), total DON (sum of free DON and DON-glucuronides or DON-GlcA) and de-epoxy deoxynivalenol (DOM-1) were analysed using a validated LC-MS/MS methodology. Total DON above the limit of quantification 0.25 ng/mL was detected in the urine from 90% of elderly men and women on both days. Mean total DON concentrations on day 1 were not different from those on day 2 (elderly men, day 1: 22.2 ± 26.3 ng/mg creatinine (creat), day 2: 28.0 ± 34.4 ng/mg creat, p = 0.95; elderly women, day 1: 22.4 ± 14.6 ng/mg creat, day 2: 29.1 ± 22.8 ng/mg creat, p = 0.58). Free DON and DON-GlcA were detected in 60-70% and 90% of total urine samples, respectively. DOM-1 was absent from all samples; the LoQ for DOM-1 was 0.50 ng/mL. Estimated dietary intake of DON suggested that 10% of the elderly exceeded the maximum provisional tolerable daily intake for DON. In this single-site, UK-based cohort, elderly were frequently exposed to DON, although mean total DON concentrations were reported at moderate levels. Future larger studies are required to investigate DON exposure in elderly from different regions of the UK, but also from different counties worldwide.

Assessment of Urinary Deoxynivalenol Biomarkers in UK Children and Adolescents.

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.

Deoxynivalenol Biomarkers in the Urine of UK Vegetarians.

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).

Study on the Association among Mycotoxins and other Variables in Children with Autism.

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.

Development of a LC-MS/MS Method for the Multi-Mycotoxin Determination in Composite Cereal-Based Samples.

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.

Survey on Urinary Levels of Aflatoxins in Professionally Exposed Workers.

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.

Determination of Deoxynivalenol in the Urine of Pregnant Women in the UK.

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.

Exposure assessment for Italian population groups to deoxynivalenol deriving from pasta consumption.

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.

Effect of sample size in the evaluation of "in-field" sampling plans for aflatoxin B(1) determination in corn.

Use of proper sampling methods throughout the agri-food chain is crucial when it comes to effectively detecting contaminants in foods and feeds. The objective of the study was to estimate the performance of sampling plan designs to determine aflatoxin B(1) (AFB(1)) contamination in corn fields. A total of 840 ears were selected from a corn field suspected of being contaminated with aflatoxin. The mean and variance among the aflatoxin values for each ear were 10.6 mug/kg and 2233.3, respectively. The variability and confidence intervals associated with sample means of a given size could be predicted using an equation associated with the normal distribution. Sample sizes of 248 and 674 ears would be required to estimate the true field concentration of 10.6 mug/kg within +/-50 and +/-30%, respectively. Using the distribution information from the study, operating characteristic curves were developed to show the performance of various sampling plan designs.