BACKGROUND: The essential oils (EO) carvacrol, thymol, eugenol and trans-cinnamaldehyde have welldocumented antimicrobial properties and offer therefore an alternative for the antimicrobial growth promoters in pig feeds. The aim of this work was to determine the degradation and kinetics of these EO along the gastrointestinal tract (GIT) of piglets, which is necessary information for correct application in pig feeds.

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

A total of 174 cereal-based food products, 67 compound feeds and 19 feed raw materials were analysed for the occurrence of deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, zearalenone, α-zearalenol, β-zearalenol, and their respective masked forms, including deoxynivalenol-3-glucoside, zearalenone-4-glucoside, α-zearalenol-4-glucoside, β-zearalenol-4-glucoside and zearalenone-4-sulfate. Fibre-enriched bread, bran-enriched bread, cornflakes, popcorn and oatmeal were collected in Belgian supermarkets from April 2010 to October 2011. All food samples analysed were contaminated with an average of 2 to 6 mycotoxins, including 1 to 3 masked forms. Feed raw materials that were used in the analysed compound feeds were collected by the manufacturer. Feed raw materials included were beet pulp, sunflower seed meal, soy bean, soy peel, oats, barley, maize germs, maize gluten feed, maize, wheat gluten feed, wheat bran pellets, wheat bran and wheat. Beet pulp, sunflower seed meal, soy bean and soy peel were hardly contaminated. The feed raw materials that were mostly infected with deoxynivalenol, zearalenone and derivatives were maize and its by-products. Also, the glucosylated and sulfated forms occurred in substantial amounts. As well, wheat and its by-products were contaminated with α-zearalenol (wheat gluten feed and wheat bran) and zearalenone (wheat). The contamination pattern and level of feed raw materials were reflected in the corresponding compound feeds.

In the present study, a quantitative dietary exposure assessment of mycotoxins and their masked forms was conducted on a national representative sample of the Belgian population using the contamination data of cereal-based foods. Cereal-based food products (n = 174) were analysed for the occurrence of deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, zearalenone, α-zearalenol, β-zearalenol, T-2-toxin, HT-2-toxin, and their respective masked forms, including, deoxynivalenol-3-glucoside, zearalenone-4-glucoside, α-zearalenol-4-glucoside, β-zearalenol-4-glucoside and zearalenone-4-sulfate.Fiber-enriched bread, bran-enriched bread, breakfast cereals, popcorn and oatmeal were collected in Belgian supermarkets according to a structured sampling plan and analysed during the period from April 2010 till October 2011. The habitual intake of these food groups was estimated from a national representative food intake survey. According to a probabilistic exposure analysis, the mean (and P95) mycotoxin intake for the sum of the deoxynivalenolequivalents, zearalenone-equivalents, and the sum of HT-2-and T-2-toxin for all cereal-based foods was 0.1162 (0.4047, P95), 0.0447 (0.1568, P95) and 0.0258 (0.0924, P95) µg kg -1 body weight day -1 , respectively. These values were below the tolerable daily intake (TDI) levels for deoxynivalenol, zearalenone and the sum of T-2 and HT-2 toxin (1.0, 0.25 and 0.1 µg kg -1 body weight day -1 , respectively). The absolute level exceeding the TDI for all cereal-based foods was calculated, and recorded 0.85%, 2.75% and 4,11% of the Belgian population, respectively. KEYWORDS: mycotoxins,

Deoxynivalenol (DON) content and Fusarium spp. (Fusarium graminearum, Fusarium culmorum, Fusarium avenaceum, Microdochium nivale and Fusarium poae) of mycotoxin-producing Fusarium species in winter wheat in Belgium (Flanders) were determined. Field trials were set up in the varietal testing network of Flanders Agricultural Centre for Small Grains (RoeselareBeitem, Belgium) and followed up during growing seasons

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Maize ear rot caused by several Fusarium species is an important fungal disease. Apart from yield losses, ear rot fungi can produce mycotoxins and masked forms in infected grains. Masked mycotoxins have received increased attention in view of their bioavailability and potential toxicity in animals and humans, but their presence and relevance in the field still remain undisclosed. To get a better insight, the present study assessed the presence of various Fusarium parent and masked mycotoxins, i.e. deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deoxynivalenol-3-glucoside, zearalenone, alpha-zearalenol, beta-zearalenol, zearalenone-14-glucoside, zearalenone-14-sulfate, alpha-zearalenol-14-glucoside, beta-zearalenol-14-glucoside, T-2 and HT-2 toxin, in various commercial maize varieties grown under natural infection conditions in Flanders, Belgium. The results showed that the maize varieties were co-contaminated with both parent and masked mycotoxins. Moreover, a positive correlation between these forms was established. A higher contamination with a particular mycotoxin appeared to be coupled with an elevated load of another (masked) mycotoxin. The results highlight the importance to screen for multiple mycotoxins, both parent and masked, to guarantee food and feed safety. Furthermore, analysis was carried out to elucidate the distribution of the various mycotoxins in the field. The maize variety did not significantly influence mycotoxin accumulation, except for deoxynivalenol. Subdivisions in the field with higher mycotoxin levels for deoxynivalenol and its derivatives, zearalenone and its derivatives, and the sum of T-2 and HT-2 toxin were observed

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