Geert Haesaert

Natural occurrence of mycotoxins and their masked forms in food and feed products

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 w...

Development and validation of an LC-MS/MS method for the simultaneous determination of deoxynivalenol, zearalenone, T-2-toxin and some masked metabolites in different cereals and cereal-derived food

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.

Human exposure to mycotoxins and their masked forms through cereal-based foods in Belgium

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. Fibre-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 to 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 deoxynivalenol-equivalents, 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.

Hydrogen peroxide induced by the fungicide prothioconazole triggers deoxynivalenol (DON) production by Fusarium graminearum

BackgroundFusarium head blight is a very important disease of small grain cereals with F. graminearum as one of the most important causal agents. It not only causes reduction in yield and quality but from a human and animal healthcare point of view, it produces mycotoxins such as deoxynivalenol (DON) which can accumulate to toxic levels. Little is known about external triggers influencing DON production.ResultsIn the present work, a combined in vivo/in vitro approach was used to test the effect of sub lethal fungicide treatments on DON production. Using a dilution series of prothioconazole, azoxystrobin and prothioconazole + fluoxastrobin, we demonstrated that sub lethal doses of prothioconazole coincide with an increase in DON production 48 h after fungicide treatment. In an artificial infection trial using wheat plants, the in vitro results of increased DON levels upon sub lethal prothioconazole application were confirmed illustrating the significance of these results from a practical point of view. In addition, further in vitro experiments revealed a timely hyperinduction of H2O2 production as fast as 4 h after amending cultures with prothioconazole. When applying H2O2 directly to germinating conidia, a similar induction of DON-production by F. graminearum was observed. The effect of sub lethal prothioconazole concentrations on DON production completely disappeared when applying catalase together with the fungicide.ConclusionsThese cumulative results suggest that H2O2 induced by sub lethal doses of the triazole fungicide prothioconazole acts as a trigger of DON biosynthesis. In a broader framework, this work clearly shows that DON production by the plant pathogen F. graminearum is the result of the interaction of fungal genomics and external environmental triggers.

Environmental impact of herbicide regimes used with genetically modified herbicide-resistant maize

With the potential advent of genetically modified herbicide-resistant (GMHR) crops in the European Union, changes in patterns of herbicide use are predicted. Broad-spectrum, non-selective herbicides used with GMHR crops are expected to substitute for a set of currently used herbicides, which might alter the agro-environmental footprint from crop production. To test this hypothesis, the environmental impact of various herbicide regimes currently used with non-GMHR maize in Belgium was calculated and compared with that of possible herbicide regimes applied in GMHR maize. Impacts on human health and the environment were calculated through the pesticide occupational and environmental risk (POCER) indicator. Results showed that the environmental impact of herbicide regimes solely relying on the active ingredients glyphosate (GLY) or glufosinate-ammonium (GLU) is lower than that of herbicide regimes applied in non-GMHR maize. Due to the lower potential of GLY and GLU to contaminate ground water and their lower acute toxicity to aquatic organisms, the POCER exceedence factor values for the environment were reduced approximately by a sixth when GLY or GLU is used alone. However, the environmental impact of novel herbicide regimes tested may be underestimated due to the assumption that active ingredients used with GMHR maize would be used alone. Data retrieved from literature suggest that weed control efficacy is increased and resistance development delayed when GLY or GLU is used together with other herbicides in the GMHR system. Due to the partial instead of complete replacement of currently used herbicide regimes, the beneficial environmental impact of novel herbicide regimes might sometimes be reduced or counterbalanced. Despite the high weed control efficacy provided by the biotechnology-based weed management strategy, neither indirect harmful effects on farmland biodiversity through losses in food resources and shelter, nor shifts in weed communities have been demonstrated in GMHR maize yet. However, with the increasing adoption rate of GMHR maize and their associated novel herbicide regimes, this situation is expected to change in the short-term.

Application of Humic Substances Results in Consistent Increases in Crop Yield and Nutrient Uptake

ABSTRACT The effect of humic substances on yield and nutrient uptake of grass, maize, potato and spinach was investigated in six field experiments and two pot experiments in a high input cropping system. The humic substances originating from leonardite formations in Canada were applied as liquid solution to the soil (Humifirst liquid) or as a solid incorporated in mineral fertilizers (Humifirst incorporated). Formal meta-analysis of the results of all executed experiments showed that the application of humic substances had an overall positive effect on dry matter yield of the crops and this effect was statistically significant for Humifirst incorporated. In the case of permanent grassland, humic substances promoted mainly the production of the first grass cut, which has the highest grass quality among all cuts during the growing season. Tuber production on the potato field trial showed a high response on the application of humic substances. Total potato yield increased with 13 and 17% for Humifirst liquid and Humifirst incorporated, respectively. The effect of humic substances on maize yield was limited, probably due to the rather high nutrient status of both soils. Finally, the formal meta-analysis showed a consequent increase in nitrogen and phosphorus uptake of all studied crops as well. The effect on potassium and magnesium uptake was also mainly positive, while sodium and calcium uptake were not affected in most of the experiments.

Support vector machine regression for the prediction of maize hybrid performance

Accurate prediction of the phenotypical performance of untested single-cross hybrids allows for a faster genetic progress of the breeding pool at a reduced cost. We propose a prediction method based on ɛ-insensitive support vector machine regression (ɛ-SVR). A brief overview of the theoretical background of this fairly new technique and the use of specific kernel functions based on commonly applied genetic similarity measures for dominant and co-dominant markers are presented. These different marker types can be integrated into a single regression model by means of simple kernel operations. Field trial data from the grain maize breeding programme of the private company RAGT R2n are used to assess the predictive capabilities of the proposed methodology. Prediction accuracies are compared to those of one of today’s best performing prediction methods based on best linear unbiased prediction. Results on our data indicate that both methods match each other’s prediction accuracies for several combinations of marker types and traits. The ɛ-SVR framework, however, allows for a greater flexibility in combining different kinds of predictor variables.

Influence of Mycotoxins and a Mycotoxin Adsorbing Agent on the Oral Bioavailability of Commonly Used Antibiotics in Pigs

It is recognized that mycotoxins can cause a variety of adverse health effects in animals, including altered gastrointestinal barrier function. It is the aim of the present study to determine whether mycotoxin-contaminated diets can alter the oral bioavailability of the antibiotics doxycycline and paromomycin in pigs, and whether a mycotoxin adsorbing agent included into diets interacts with those antibiotics. Experiments were conducted with pigs utilizing diets that contained blank feed, mycotoxin-contaminated feed (T-2 toxin or deoxynivalenol), mycotoxin-contaminated feed supplemented with a glucomannan mycotoxin binder, or blank feed supplemented with mycotoxin binder. Diets with T-2 toxin and binder or deoxynivalenol and binder induced increased plasma concentrations of doxycycline administered as single bolus in pigs compared to diets containing blank feed. These results suggest that complex interactions may occur between mycotoxins, mycotoxin binders, and antibiotics which could alter antibiotic bioavailability. This could have consequences for animal toxicity, withdrawal time for oral antibiotics, or public health.

Higher sustainability performance of intensive grazing versus zero-grazing dairy systems

Although grazing of dairy cows is an integral part of dairy farming in many European countries, farmers today more often choose for zero-grazing systems, where cows are housed throughout the year. Some studies already compared grazing and zero-grazing systems for specific issues such as labor efficiency, environmental impact, or animal welfare. In our study, we perform a more integrated evaluation, considering relevant ecological, economic, and social aspects. This allows for a balanced and more complete comparison of the sustainability performance of the two production methods. We evaluated ten intensive grazing and ten zero-grazing specialized Flemish dairy farms on the use of nutrients and energy, productivity and profitability, labor input, and animal welfare. In addition, we put special effort in formulating useful management advice for farmers. Therefore, we combined a detailed analysis of the sustainability indicators with an intensive interaction and discussion with farmers and farm advisors. Results show that, on average, the zero-grazing farms performed significantly worse from an ecological and economic point of view. This fact is explained mainly due to a less efficient use of concentrates and byproducts. Social sustainability performance did not differ significantly between the two groups. As a result, the integrated sustainability performance was significantly lower for the zero-grazing group. This finding shows that a further shift from intensive grazing to zero-grazing can move dairy farming in Flanders further away from sustainability. An important advice to improve the ecological and economic performance of zero-grazing farms is to optimize cows’ rations to include more forages and optimize forage production and use. More detailed site- and case-specific management advice for farmers of both groups was provided during a discussion meeting. We consider this an essential additional step to any sustainability evaluation, since progress can only be made when monitoring results are translated into practical measures.

Prediction of maize single-cross hybrid performance: support vector machine regression versus best linear prediction

Accurate prediction of the phenotypic performance of a hybrid plant based on the molecular fingerprints of its parents should lead to a more cost-effective breeding programme as it allows to reduce the number of expensive field evaluations. The construction of a reliable prediction model requires a representative sample of hybrids for which both molecular and phenotypic information are accessible. This phenotypic information is usually readily available as typical breeding programmes test numerous new hybrids in multi-location field trials on a yearly basis. Earlier studies indicated that a linear mixed model analysis of this typically unbalanced phenotypic data allows to construct ɛ-insensitive support vector machine regression and best linear prediction models for predicting the performance of single-cross maize hybrids. We compare these prediction methods using different subsets of the phenotypic and marker data of a commercial maize breeding programme and evaluate the resulting prediction accuracies by means of a specifically designed field experiment. This balanced field trial allows to assess the reliability of the cross-validation prediction accuracies reported here and in earlier studies. The limits of the predictive capabilities of both prediction methods are further examined by reducing the number of training hybrids and the size of the molecular fingerprints. The results indicate a considerable discrepancy between prediction accuracies obtained by cross-validation procedures and those obtained by correlating the predictions with the results of a validation field trial. The prediction accuracy of best linear prediction was less sensitive to a reduction of the number of training examples compared with that of support vector machine regression. The latter was, however, better at predicting hybrid performance when the size of the molecular fingerprints was reduced, especially if the initial set of markers had a low information content.