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Mycotoxins are natural toxins produced by fungi that may cause adverse health effects thus constituting a public health concern. Deoxynivalenol (DON), a mycotoxin affecting the immune system and causing intestinal disorders, was selected as a priority under the European Human Biomonitoring Initiative (HBM4EU). Urinary total DON levels (tDON) of 1270 participants from six countries were used to characterize the internal exposure of the adult European population and identify the most relevant determinants of exposure. tDON concentrations' P50 and P95 were in the range of 0.41-10.16 µg/L (0.39-9.05 µg/g crt) and 3.25-46.58 µg/L (2.12-33.50 µg/g crt) respectively. Higher tDON levels were observed for (i) male participants from France and Germany, (ii) samples collected in spring and summer, (iii) participants with a lower educational level, (iv) participants living in rural areas, (v) individuals without a job in France and Luxembourg, while in Portugal higher exposure was observed in working individuals, (vi) individuals with higher consumption of cereals and bread. The proportion of individuals with exposure levels exceeding the HBM-GV of 23 µg/L was 12.3 %, ranging from 0.8 % to 20.7 % in the individual countries. This study on mycotoxins exposure has used post harmonized questionnaire data and validated analytical methodologies for analysis and covered countries representing the four geographical regions of Europe, having produced much needed knowledge on the exposure of the European adult population to deoxynivalenol., 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 Ltd.. All rights reserved.)

The widespread presence of Fusarium mycotoxins in animal feed is a global issue, not only for the health of livestock but also for ensure the safety of food as an end product. High concentrations of zearalenone (ZEN) and deoxynivalenol (DON) have been detected in the diets of Japanese Black (JB) and Holstein Friesian (HF) breeding herds. Consequently, we monitored serum biochemical parameters over a long time in both herds, focusing on anti-Müllerian hormone (AMH) levels and acute-phase inflammation. Additionally, urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) and progesterone levels were measured in the HF herd. The JB herd, a ZEN-dominant model with low DON contamination, demonstrated ZEN levels that exceeded the Japanese limit in the purchased total mixed rations (TMR). Conversely, the HF herd, which primary consumes DON-dominant feed with low ZEN contamination, had high DON levels in the dent corn silage. Specifically, the JB herd's TMR contained 1.79 mg/kg ZEN and 0.58 mg/kg DON, whereas the HF herd's silage had 15.3 mg/kg DON (dried sample) and 0.1 mg/kg ZEN. Enzyme-linked immunoassay were used to measure urinary ZEN-DON levels following confirmation through liquid chromatography-tandem mass spectrometry. Urinary ZEN-DON levels measured were significantly correlated ( p < 0.05, r > 0.6) in both herds. In the HF herd, AMH levels increased ( p = 0.01) and serum amyloid A (SAA) levels decreased ( p = 0.02) when contaminated and at the end of the monitoring period. Additionally, urinary ZEN and DON levels were significantly correlated with SAA levels (ZEN: p = 0.00, r = 0.46; DON: p = 0.03, r = 0.33), with an increase in ZEN and DON levels resulting in higher SAA levels. The JB herd showed no significant differences. Additionally, in the HF herd, 8-OHdG/Cre levels increased significantly during major contamination periods ( p < 0.05). Clinical data from the HF herd indicated an increase in mastitis cases and treatment rates during periods of major contamination. Abortion rates in the HF herd decreased from 22.9% (before monitoring) to 8.9% (during the high contamination period) and finally to 1% (at the end of the monitoring period), with corresponding increases in progesterone levels. ZEN-DON contamination adversely affects breeding cattle's productivity, reproductive performance, and health. Therefore, monitoring urinary ZEN-DON is valuable for detecting contaminants and ensuring the safety of food products.

Deoxynivalenol is one of the most ubiquitous mycotoxins in the Western diet through its presence in cereals and cereal products. A vast amount of studies indicate the worrying level of exposure to this toxin, while even high percentages of the population exceed the tolerable daily intake. To evaluate and assess dietary exposure, analysis of urinary levels of deoxynivalenol and its glucuronides has been proposed as a reliable methodology. An indirect preliminary method was used based on the cleavage of deoxynivalenol glucuronides through the use of enzymes (β-glucuronidase) and subsequent determination of "total deoxynivalenol" (sum of free and released mycotoxins by hydrolysis). Next, a direct procedure for quantification of deoxynivalenol-3-glucuronide and deoxynivalenol-15-glucuronide was developed. As deoxynivalenol glucuronides reference standards are not commercially available, the indirect method is widely applied. However, to not underestimate the total deoxynivalenol exposure in urine, the direct and indirect methodologies need to be compared. Urinary samples (n = 96) with a confirmed presence of deoxynivalenol and/or deoxynivalenol glucuronides were analysed using both approaches. The indirect method clarified that not all deoxynivalenol glucuronides were transformed to free deoxynivalenol during enzymatic treatment, causing an underestimation of total deoxynivalenol. This short communication concludes on the application of direct or indirect assessment of urinary deoxynivalenol., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Deoxynivalenol (DON) exposure is estimated by the combined measures of urinary DON and DON-glucuronides. In this study, data from single-mycotoxin (SM) and a multimycotoxin (MM) methods were compared for 256 Swedish adult urine samples. Both methods included β-glucuronidase predigestion, immunoaffinity enrichment, and LC-MS/MS. However, the specific reagents, apparatus, and conditions were not identical in part because the MM method measures additional mycotoxins. DON was detected in 88 and 63% of samples using the SM and MM methods, respectively, with the following mean and median concentrations: SM, mean = 5.0 ng/mL, SD = 7.4, range of positives = 0.5-60.2 ng/mL, median = 2.5 ng/mL, IQR = 1.0-5.5 ng/mL; MM, mean = 4.4 ng/mL, SD = 12.9, range of positives = 0.5-135.2 ng/mL, median = 0.8 ng/mL, IQR = 0.3-3.5. Linear regression showed a significant, albeit modest, correlation between the two measures (p = 0.0001, r = 0.591). The differences observed may reflect subtle handling differences in DON extraction and quantitation between the methods.

Deoxynivalenol (DON) is a frequent mycotoxin contaminant in cereal crops worldwide and can cause adverse health effects in exposed animals and humans. Since DON contamination in Bangladeshi food is unexplored, we conducted a biomonitoring study to assess DON exposure in the Bangladeshi population and compare it with that of German adults. In total 214 urines were collected, n=164 in Bangladesh and n=50 in Germany. In Bangladesh rural and urban residents of Rajshahi district provided urines in two seasons (n=69 in summer, n=95 in winter, with 62 participants enrolled in both periods). Urinary DON and its de-epoxy metabolite DOM-1 were measured by a previously validated sensitive LC-MS/MS method. In Bangladeshi urines, DON was detectable in 27% (range 0.16-1.78ng/mL) in summer and 31% (range 0.16-1.21ng/mL) in winter season. There was no significant difference at the mean DON level between season (summer 0.17±0.25ng/mL and winter 0.16±0.18ng/mL) and region (rural or urban residents). The metabolite DOM-1 was not detected in any urine from Bangladesh. In contrast, DON and DOM-1 were detected in 100% (range 0.16-38.44ng/mL) and 40% (range 0.10-0.73ng/mL), respectively, of the German urines. The mean DON level in German urines (9.02±6.84ng/mL) was about 53-fold higher than that found in Bangladeshi samples. This indicates a low and high dietary DON exposure among the adult population in Bangladesh and Germany, respectively. The biomarker concentrations found and published urinary excretion rates for DON then served to calculate the daily mycotoxin intake in both cohorts: the mean DON intake in Bangladesh being 6ng/kg b.w., and in Germany a mean of 268 and maximum intake of 975ng/kg b.w., values lower than the provisional maximum tolerable daily intake of 1μg/kg b.w. set by the WHO/JECFA., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Mycotoxins are important food contaminants responsible for health effects such as cancer, nephrotoxicity, hepatotoxicity or immunosuppression. The assessment of mycotoxin exposure is often based on calculations combining mycotoxin occurrence data in food with population data on food consumption. Because of limitations inherent to that approach, the direct measurement of biomarkers of exposure in biological fluids has been proposed as a suitable alternative to perform an accurate mycotoxin exposure assessment at individual level. For this reason, the BIOMYCO study was designed to assess mycotoxin exposure in Belgian adults and children using urinary biomarkers of exposure. Morning urine was gathered in a representative part of the Belgian population according to a standardised study protocol, whereby 155 children (3-12 years old) and 239 adults (19-65 years old) were selected based on random cluster sampling. These urine samples were analysed for the presence of 33 potential biomarkers with focus on aflatoxins, citrinin (CIT), fumonisins, trichothecenes, ochratoxin A (OTA), zearalenone and their metabolites using two validated LC-MS/MS methods. Nine out of the 33 analysed mycotoxins were detected whereby deoxynivalenol (DON), OTA, CIT and their metabolites were the most frequently detected. Deoxynivalenol-15-glucuronide was the main urinary DON biomarker and was found in all urine samples in the ng/mL range. Furthermore deoxynivalenol-3-glucuronide was quantified in 91% of the urine samples collected from children and in 77% of the samples collected from adults. Deoxynivalenol was detected in 70% and 37% of the samples of children and adults respectively. For the first time deepoxy-deoxynivalenol-glucuronide was detected in children's urine (17%). In the samples collected by adults, the prevalence was 22%. Whereas all these mycotoxins contaminated the urine samples in the ng/mL range, CIT and OTA were present in much lower concentrations (pg/mL). OTA contaminated 51% and 35% of the samples collected by children and adults respectively. CIT and its metabolite were present in 72% and 6% of children's urine, whereas they contaminated 59% and 12% of adult's urine. Finally, α-zearalenol and β-zearalenol-14-glucuronide were found in respectively one and two samples from adults. The exposure to DON, OTA and CIT was compared between subgroups and urinary mycotoxin concentrations differed significantly among age and gender. Based on the urinary levels, the daily intake of DON and OTA was estimated and evaluated whereby, depending on the used method, 16-69% of the population possibly exceeded the tolerable daily intake for DON and 1% for OTA. The BIOMYCO study is the first study whereby a multi-toxin approach was applied for mycotoxin exposure assessment in adults and children on a large-scale. Moreover, it is the first study that described the exposure to an elaborated set of mycotoxins in the Belgian population. Biomarker analysis showed a clear exposure of a broad segment of the Belgian population to DON, OTA and CIT. The risk assessment based on these data indicates a potential concern for a number of individuals whereby young children need special attention because of the relatively higher food intake per kg body weight., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Mycotoxin producing moulds may contaminate numerous agricultural commodities either before harvest or during storage. A varied diet consisting of different foods may therefore be contaminated with a range of mycotoxins. The aim of the present study was to study concurrent exposure to mycotoxins through urinary multi-biomarker analysis, as well as its possible associations with the diet. Urinary samples from 252 adults, participating in the Swedish national dietary survey Riksmaten 2010-11, were collected together with a 4-day diet record. Concurrent mycotoxin exposure was studied using a multi-biomarker LC-MS/MS method. The results revealed that exposure to mycotoxins is common and concurrent exposure to more than one toxin was found in 69% of the study population. However, when comparing the number of toxins detected with the reported consumption data it was difficult to distinguish food patterns which would indicate an increased risk of exposure to many mycotoxins simultaneously. This is the first study to investigate concurrent mycotoxin exposure and urinary levels of fumonisin B1 (FB1), fumonisin B2 (FB2), nivalenol (NIV), ochratoxin A (OTA), zearalenone (ZEA), α-zearalenol (α-ZOL), β-zearalenol (β-ZOL) and de-epoxydeoxynivalenol (DOM-1) among adults in Sweden., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

A duplicate diet study was designed to explore the occurrence of 15 Fusarium mycotoxins in the 24 h-diet consumed by one volunteer as well as the levels of mycotoxins in his 24 h-collected urine. The employed methodology involved solvent extraction at high ionic strength followed by dispersive solid phase extraction and gas chromatography determination coupled to mass spectrometry in tandem. Satisfactory results in method performance were achieved. The method's accuracy was in a range of 68%-108%, with intra-day relative standard deviation and inter-day relative standard deviation lower than 12% and 15%, respectively. The limits of quantitation ranged from 0.1 to 8 µg/Kg. The matrix effect was evaluated and matrix-matched calibrations were used for quantitation. Only deoxynivalenol (DON) was quantified in both food and urine samples. A total DON daily intake amounted to 49.2 ± 5.6 µg whereas DON daily excretion of 35.2 ± 4.3 µg was determined. DON daily intake represented 68.3% of the established DON provisional maximum tolerable daily intake (PMTDI). Valuable preliminary information was obtained as regards DON excretion and needs to be confirmed in large-scale monitoring studies.

Scope: This study assessed deoxynivalenol (DON) exposure in children from three geographic locations within Tanzania, over three time points in 1 year, using a urinary biomarker of exposure., Methods and Results: A total of 166 children aged 6-14 months were studied at a maize harvest and followed up twice at 6-month intervals. On two consecutive days, morning urine was collected from each child and urinary DON was measured using an LC-MS method, with and without β-glucuronidase hydrolysis in order to assess free DON (fDON) and glucuronide DON. Overall, urinary DON increased significantly along with the three visits (geometric mean 1.1, 2.3, and 5.7 ng/mL, at visits 1, 2, and 3, respectively, p < 0.01). fDON was 22% of urinary total DON. Urinary DON excretion rate was 74% in village Kikelelwa based on food DON level and food consumption. Assuming 360 mL of urine excreted per day, 10, 19, and 29% of children at visits 1, 2, and 3, respectively, exceeded the provisional maximum tolerable daily intake of 1000 ng/kg b.w./day., Conclusion: Young children in Tanzania are chronically exposed to DON due to eating contaminated maize, although exposure levels varied markedly by region and season., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Human exposures to mycotoxins through dietary intake are a major health hazard and may result in various pathophysiological effects. Although Thailand is a country at increased risk due to its climatic conditions, no comprehensive dataset is available to perform proper exposure assessment of its population with regard to mycotoxins. Therefore, this pilot study was conducted to investigate and evaluate the exposure levels of major mycotoxins (aflatoxin B₁, ochratoxin A, fumonisins, zearalenone and trichothecenes). Sixty first-morning urine samples were collected from healthy volunteers who live in the Bangkok metropolitan area and surrounding provinces (Pathumthani, Nonthaburi, Samutprakarn and Samutsakorn). Urine samples were analysed by a LC-MS/MS-based multi-biomarker method following a so-called 'dilute and shoot' approach. Results generally indicated low mycotoxin exposures in most individuals through the determination of the four biomarkers that were detected in urine samples, i.e. aflatoxin M₁, ochratoxin A (OTA), as well as the deoxynivalenol (DON) metabolites DON-3-glucuronide and DON-15-glucuronide in 10 of 60 individuals. The maximum concentrations were used to estimate the daily intake confirming that none of the individuals exceeded the tolerable daily intake (TDI) of DON (maximum 26% of TDI) or OTA (maximum 22% of TDI). However, the maximum exposure of aflatoxin B₁, estimated to be 0.91 µg (kg bw)⁻¹ day⁻¹, should raise some concerns and suggests further studies utilising a more sensitive method. Low exposure to Fusarium toxins was also confirmed by the absence of zearalenone, α-zearalanol, β-zearalanol and zearalenone-14-glucuronide as well as T-2 toxin, HT-2 toxin, nivalenol and free DON. This is the first multi-mycotoxin biomarker study performed in Southeast Asia.

Subsistence farmers are exposed to a range of mycotoxins. This study applied novel urinary multi-mycotoxin LC-MS/MS methods to determine multiple exposure biomarkers in the high oesophageal cancer region, Transkei, South Africa. Fifty-three female participants donated part of their maize-based evening meal and first void morning urine, which was analysed both with sample clean-up (single and multi-biomarker) and by a 'dilute-and-shoot' multi-biomarker method. Results were corrected for recovery with LOD for not detected. A single biomarker method detected fumonisin B1 (FB1) (87% incidence; mean±standard deviation 0.342±0.466 ng/mg creatinine) and deoxynivalenol (100%; mean 20.4±49.4 ng/mg creatinine) after hydrolysis with β-glucuronidase. The multi-biomarker 'dilute-and-shoot' method indicated deoxynivalenol-15-glucuronide was predominantly present. A multi-biomarker method with β-glucuronidase and immunoaffinity clean-up determined zearalenone (100%; 0.529±1.60 ng/mg creatinine), FB1 (96%; 1.52±2.17 ng/mg creatinine), α-zearalenol (92%; 0.614±1.91 ng/mg creatinine), deoxynivalenol (87%; 11.3±27.1 ng/mg creatinine), β-zearalenol (75%; 0.702±2.95 ng/mg creatinine) and ochratoxin A (98%; 0.041±0.086 ng/mg creatinine). These demonstrate the value of multi-biomarker methods in measuring exposures in populations exposed to multiple mycotoxins. This is the first finding of urinary deoxynivalenol, zearalenone, their conjugates, ochratoxin A and zearalenols in Transkei., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The aim of this study was to examine the effects of a control diet (CON, 0.25 mg DON/kg diet) or a Fusarium toxin-contaminated diet (FUS, 4.49 mg DON/kg diet) without and with humic substances (HS) (CON-HS and FUS-HS, 0.23 and 4.56 mg DON/kg diet, respectively) on piglets during a 5-week growth trial starting after weaning (6.7 ± 0.9 kg live weight, n = 20/group). Feed intake was significantly reduced by feeding the FUS containing diets by approximately 21% compared with the CON diet irrespective of HS supplementation. The decrease in live weight gain paralleled the feed intake depression and amounted to approximately 26%. Feeding the FUS diet was clearly reflected by the DON levels in blood. While only traces of DON with median concentrations of 3 ng/ml (2-5 ng/ml) and 2 ng/ml (0-3 ng/ml) were detected in piglets fed the CON and CON-HS diets, respectively, significantly higher levels of 22.5 ng/ml (7-30 ng/ml) and 23.5 ng/ml (15-32 ng/ml) were found in piglets fed the FUS and FUS-HS diet, respectively. The urinary excretion of DON and its metabolite de-epoxy-DON as percentage of DON intake was not significantly influenced by HS supplementation and amounted to 24.1 and 20.2% for groups FUS and FUS-HS, respectively. In conclusion, the tested HS preparation cannot be recommended as a DON inactivating feed supplement for pigs.

Deoxynivalenol (DON) is a ubiquitous contaminant of cereal crops in temperate regions of the world. It causes growth faltering and immune suppression in animals. Limited information is available on DON exposure in UK subpopulations. The objective of this study was to provide DON exposure assessment in a subset of pregnant women scheduled for an elective caesarean in a large multi-ethnic mother/infant birth cohort from Bradford, UK. Women aged 16-44 years (n = 85) provided a urine sample for DON analysis in the last trimester of pregnancy, and concurrently completed a food-frequency questionnaire (FFQ). The urinary DON biomarker was detected in all measured samples (geometric mean (GM) = 10.3 ng DON mg(-1) creatinine, range = 0.5-116.7 ng mg(-1)). Levels were higher in women classified as South Asian in origin (GM: 15.2 ng mg(-1); 95% CI = 10.7-21.5 ng mg(-1)) compared with non-South Asians (GM = 8.6 ng mg(-1); 95% CI = 6.6-11.8 ng mg(-1)), p = 0.02). Estimated DON intake from FFQ data and typical levels of DON contamination of food suggest that this was mainly due to higher levels of exposure from bread, particularly daily intake of DON from chapattis in South Asians (estimated mean = 2.4 µg day(-1); 95% CI = 1.2, 3.7 µg day(-1)) compared with non-South Asians (estimated mean = 0.2 µg day(-1); 95% CI = 0-0.4 µg day(-1)), p < 0.001. This is the first biomarker demonstration of DON exposure in pregnant women, and several urinary DON levels were the highest ever recorded in any study. A larger survey within this birth cohort is warranted to investigate any potential risk to mothers and their babies, from DON exposure during pregnancy.

The Fusarium toxin deoxynivalenol (DON) is of outstanding importance in pig nutrition because of its frequent occurrence in cereal grains at levels high enough to cause adverse effects such as a decrease in feed intake and impairment of the immune system. Thus, simple decontamination procedures would be useful. The present study aimed to examine the effects of wet preservation of triticale contaminated with DON and zearalenone (ZON) with sodium metabisulphite (SBS) on the treatment-related non-toxic derivative of DON (DON-sulfonate, DONS), and on ZON and its metabolites in blood and various physiological specimens of piglets. The uncontaminated control triticale (CON) and the DON-contaminated triticale (FUS) were included in the diets either untreated or SBS treated (CON-SBS, FUS-SBS) and fed to piglets for 28 days starting from weaning. The diet concentrations for DON were 0.156, 0.084, 2.312 and 0.275 mg kg(-1), for DONS were <0.05, <0.05, <0.05 and 1.841 mg kg(-1), and for ZON were <0.001, 0.006, 0.017, and 0.016 mg kg(-1) for each of CON, CON-SBS, FUS and FUS-SBS, respectively. DONS was present in the blood of piglets fed the FUS-SBS at a median concentration of 15.5 ng ml(-1) (3-67 ng ml(-1)), while the median DON concentration amounted to 2 ng ml(-1) (0-5 ng ml(-1)) at the same time. The median DON concentration in the blood of piglets fed the FUS diet reached a median concentration of 10.5 ng ml(-1) (5-17 ng ml(-1)). Moreover, the relative differences between the DON concentrations in other physiological specimens (muscle, liver, kidney, bile and urine) in piglets fed the FUS-SBS and the FUS diet were comparable with the blood DON concentration differences. Although these differences can be taken as an indication for DONS stability after absorption and distribution further studies examining DONS in these other physiological specimens directly are necessary to substantiate this conclusion. Moreover, ZON and α-zearalenol could only be detected in bile and urine where their levels were not influenced by the SBS treatment.

The relationship between deoxynivalenol (DON) intake and first morning urinary DON was examined in UK adults to validate the latter as a biomarker of human exposure. DON was assessed in first morning samples collected during a period of normal diet, a wheat-restriction intervention diet, and partial wheat-restriction intervention in which bread was allowed. During the partial intervention duplicate bread portions were collected for DON analysis. During the normal diet, partial intervention and full intervention, urinary DON was detected in 198/210 (geometric mean 10.1 ng DON mg(-1) creatinine, 95% confidence interval (CI) 8.6-11.6 ng mg(-1); range nd-70.7 ng mg(-1)), in 94/98 (5.9 ng mg(-1), 95% CI 4.8-7.0 ng mg(-1); range nd-28.4 ng mg(-1)), and 17/40 (0.5 ng mg(-1), 95% CI 0.3-0.7 ng mg(-1); range nd-3.3 ng mg(-1)) volunteers, respectively. A strong correlation between DON intake and the urinary biomarker was observed (p <0.001, adjusted r(2) = 0.83) in models adjusting for age, sex and body mass index. These data demonstrate a quantitative correlation between DON exposure and urinary DON, and serve to validate the use of urinary DON as an exposure biomarker.

Deoxynivalenol (DON) is a toxic fungal metabolite found on wheat, maize and barley. We previously reported a significant association between the amount of DON in a single 24 h urine sample and the average cereal intake over 7 d for 300 UK adults. In this more detailed analysis of the data, food diary information (n 255) for the day of urine collection (model I), the previous 24 h period (model II) and the day of urine collection plus the previous 24 h combined (model III) were further examined to assess whether the recent intake of cereal correlated more strongly with urinary DON, compared with the longer-term assessment of usual cereal intake from 7 d food diaries (model IV). DON was detected in 254/255 (99.6 %) urine samples (mean 12.0 microg/d; range not detected-66 microg/d). For all the models, total cereal intake was positively associated with urinary DON (P < 0.001) in each model. The goodness of fit (adjusted R2 value) was used to assess how well each model explained the variation in urinary DON. Model I provided a better goodness of fit (adjusted R2 0.22) than model IV (adjusted R2 0.19), whereas model III provided the best fit (adjusted R2 0.27). These data suggest that the inter-individual variation in urinary DON was somewhat better explained by recent cereal intake compared with usual cereal intake assessed over 7 d.

Mycotoxins are common dietary contaminants in most regions of the world. The frequency of exposure to the various families of mycotoxins is often dependent on geographic location, national wealth and related agricultural and regulatory infrastructure, combined with diversity of diet and degree of food sufficiency. Deoxynivalenol (DON) is a Fusarium mycotoxin that frequently contaminates wheat, corn and barley in temperate regions. A number of acute poisoning incidences have been linked to DON-contaminated foods and chronic exposure to lower levels of DON has been predicted in many regions. DON is a potent animal toxin and exposure in humans may cause gastroenteritis, growth faltering and immune toxicity. An ability to conduct accurate exposure assessment at the individual level is required to fully understand the potential health consequences for humans. To date, such exposure biomarkers have been lacking for many important mycotoxins, including DON. To better assess exposure to DON at the individual level, we have developed a robust urinary assay, incorporating immunoaffinity column (IAC) enrichment and LC-MS detection. Further refinement of this urinary assay, by inclusion of (13)C-DON as an internal standard, was then undertaken and tested within the UK. DON was frequently observed in urine and was significantly associated with cereal intake. A dietary intervention study demonstrated that avoiding wheat in the diet markedly reduced urinary levels of DON. This biomarker requires further validation but our initial data suggest it may provide a useful tool in epidemiological investigations of the potential health consequences of this common environmental toxin.

In animals deoxynivalenol (DON) causes vomiting, feed refusal, growth retardation, and affects the immune system. DON is a common contaminant of wheat, however, validated biomarker data to assess exposure at the individual level and therefore any associated health effects are lacking. The development of a highly robust assay for urinary DON involving immunoaffinity (IAC) clean-up and liquid chromatography (LC)-mass spectrometric (MS) detection has allowed the assessment of (1) DON exposure within UK individuals and (2) a wheat intake intervention on urinary DON levels. Twenty-five volunteers from the United Kingdom (aged 21-59 years) completed semi-weighed food diaries on days 1 and 2 (normal diet), and a morning urine sample was provided on day 3. On days 3-6 (intervention), individuals restricted major sources of wheat intake following dietary guidance. Diaries were completed on days 5 and 6, and a further morning urine was provided on day 7. Urinary DON was measured following IAC clean-up and analysis by LC-MS. Wheat-based food intake (mean 322 g/day, range: 131-542 g/day), was significantly (P<0.001) reduced during the intervention to 26 g/day (range: 0-159 g/day) indicating good compliance. DON was detected in all 25 urine samples taken on day 3 (geometric mean 7.2 ng DON/mg creatinine (95% confidence interval (CI) 4.9-10.5 ng/mg), but following the intervention there was a significant 11-fold reduction (P<0.001) to 0.6 ng per mg (95% CI 0.4-0.9 ng/mg). These data are unique in demonstrating human exposure to DON in the United Kingdom using a urinary biomarker. Furthermore, the study demonstrates that exposure can be markedly reduced by avoiding wheat in the diet. On the basis of urinary biomarker levels some individuals are predicted to exceed current recommended daily intakes of DON, and thus the health consequences of these exposures merit further investigation.

This study investigates the occurrence of aflatoxins in Ecuador. Early investigators proved the presence of aflatoxins in human and animal food, but the disturbing data lead to the formation of two research teams at Guayaquil University and the Agrarian University of Ecuador to investigate aflaxotins and other mycotoxins in food and their relationship to human health. Because the concept of mycotoxicosis as a result of the secondary metabolites produced by different species of moulds could cause different clinical patterns, the research team includes Aspergillus metabolites found in the urine of a patient with pulmonary aspergilloma. We considered that the body itself could create secondary metabolites. An ELISA method was used to detect mycotoxins with the specific reactive compounds using a company base assay. This allows the detection quantitative of such metabolites in 24 h collected urine. The patient was treated with itraconazole for nine months, after clinical, radiological and aflatoxins testing. We also investigated three other cases in children with a second level of malnutrition and only with vomitoxins results and in three investigated cases of otomycosis caused by Aspergillus niger only in one case traces of aflatoxins were found.

A dose response study was carried out with pigs in order to examine the effects of increasing dietary deoxynivalenol (DON)-concentrations on performance, clinical serum characteristics, nutrient digestibility and DON-metabolism. For this purpose, wheat contaminated naturally with Fusarium toxins was incorporated into pig diets at increasing proportions to give calculated dietary DON-concentrations of 0, 2.3 and 4.6 mg/kg during the starter period of phase 1 (14 d) of the experiment, and 0/0, 1.2/1.4, 2.3/3.7 mg/kg starter/grower diet during phase 3 (56 d) of the experiment. Each diet was tested on 16 pigs of both sexes with an initial average live weight of approximately 28 kg. A recovery phase (phase 2, 21 d) was intercalated between phase 1 and 3 of the growth experiment where all groups were fed with the uncontaminated control diet since some pigs exposed to the highest dietary DON-concentration during phase 1 nearly completely refused the offered feed. Affected pigs completely recovered during this phase. In phase 3, when diets with lower DON-concentrations were fed, no differences in performance could be detected. Serum clinical characteristics (enzymes indicating liver damage, total protein, immunoglobulins) did not respond to increasing DON-concentration in the diets. DON-concentration in serum increased in a dose-response-related manner as dietary DON-concentration increased. However, this parameter was not or only weakly correlated to any of the examined performance parameters or serum characteristics. Also, nutrient digestibility of the diets and N-retention were not affected by treatments with the exception of crude fat digestibility which was not consistently influenced. Concentration of DON and its metabolite de-epoxy-DON increased in urine with increasing dietary DON-concentration in a strongly linearly related fashion. The proportion of the excretion of de-epoxy-DON of the total urinary excretion of DON plus de-epoxy-DON rose linearly up to approximately 4%. Total recovery of DON plus de-epoxy-DON as percentage of DON-intake varied between 45 and 57% and was not influenced by dietary DON-concentration. Only a very small fraction of approximately 0.1% of ingested DON was recovered in faeces.