E. K. Tangni

Patulin in domestic and imported apple-based drinks in Belgium: occurrence and exposure assessment.

Apple-based beverages are regularly consumed by adults and children in Belgium. They are locally produced or imported from other countries. The apples used as starting material for these productions are frequently contaminated by mycotoxin-producing moulds and damaged during transport and handling. The current study was undertaken to investigate whether patulin (PAT) is present in the industrial or handicraft-made apple juices and ciders consumed by the Belgian population and to assess the populations exposure to this mycotoxin through apple-based drinks. Belgian (n = 29) and imported (14) apple juices as well as ciders (7) were assayed for PAT by high-performance liquid chromatography with ultraviolet light detection. PAT was detected respectively in 79, 86 and 43% of these tested samples. However, no contaminated sample exceeded the safe level of 50 μg PAT l-1. Levels of PAT contamination were comparable in Belgian and imported juice samples. The overall mean PAT concentrations were 9.0 and 3.4 μg l-1 for contaminated apple juices and ciders, respectively. This study also indicates that there was no statistically significant difference in the mean PAT contamination between clear (7.8 μg lμ1) and cloudy (10.7 μg lμ1) apple juices, as well as between handicraft-made apple juices (14.6 μg lμ1) and industrial ones (7.0 μg lμ1). On the basis of the mean results, a consumer exposure assessment indicates that a daily intake of 0.2 litres apple juice contributes to 45% of the provisional maximum tolerable daily intake for a child of 10 kg body weight.

Comparison of ochratoxin A and deoxynivalenol in organically and conventionally produced beers sold on the Belgian market

Abstract Beer was chosen as a cereal-derived and homogeneous product for a comparison of organic and conventional production methods in terms of mycotoxin contamination levels. Ochratoxin A (OTA, a storage mycotoxin) and deoxynivalenol (DON, a field mycotoxin) were assessed by HPLC in organically and conventionally produced beers sold in Belgium. Immunoaffinity column (OchraTest® and DONPrep®) purification was used prior to HPLC analysis. For in-house validation, recovery experiments, carried out with the spiked beers in the ranges of 50–200u2009ng OTAu2009l−1 and 20–100u2009µg DONu2009l−1, led to the overall averages of 91% (RSDu2009=u200910%, nu2009=u20099) and 93% (RSDu2009=u20095%, nu2009=u200927), respectively. Organic beers collected during 2003–2004 were more frequently OTA-contaminated (95%, nu2009=u200940) than their conventional counterparts (50%, nu2009=u200940). Conventional beers were OTA-contaminated at a mean concentration of 25u2009ngu2009l−1 (range: 19–198u2009ngu2009l−1), while organic beers contained a mean level of 182u2009ngu2009l−1 (range: 18–1134u2009ngu2009l−1). High OTA contamination above the limit of 200u2009ngu2009l−1 (up to 1134u2009ngu2009l−1) occasionally occurred in organically produced beers. A complementary survey performed with the same brands in 2005 did not confirm this accidental presence of excessive OTA loads (range: 3–67u2009ngu2009l−1 for 10 conventional beers and 19–158u2009ngu2009l−1 for 10 organic beers). Establishing a maximum of 3u2009µg OTAu2009kg−1 in malt, the application of the regulation EC No. 466/2001 (entered in force before the last sampling) may be related to the observed improvement. The overall incidence of DON was 67 and 80% in conventional and organic beers, respectively. DON concentrations ranged from 2 to 22u2009µg DONu2009l−1 (meanu2009=u20096u2009µg DONu2009l−1) in conventional beers, while organic beers ranged from 2 to 14u2009µg DONu2009l−1 (mean=4u2009µg DONu2009l−1). Thus, DON in beers does not appear to be a major matter of concern. From the statistical tests, it was concluded that the variation between different batches was significant (Pu2009<u20090.0001), in contrast to that observed between different brands, showing a lack of homogeneity in the raw materials. This occurs either in organically or in conventionally produced materials. Considering these results, an optimized frequency of controls according to European Regulations EC No 466/2001 and EC No 856/2005 should be recommended to reject the irregular batches.

Assessment of the chemical contamination in home-produced eggs in Belgium: general overview of the CONTEGG study.

This overview paper describes a study conducted for the Belgian Federal Public Service of Health, Food Chain Safety and Environment during 2006-2007. Home-produced eggs from Belgian private owners of hens were included in a large study aiming to determine concentration levels of various environmental contaminants. By means of the analyses of soil samples and of kitchen waste samples, obtained from the same locations, an investigation towards the possible sources of contaminants was possible. Eggs, soils, faeces and kitchen waste samples were checked for the presence of dioxins, PCBs (including dioxin-like PCBs), organochlorine pesticides, trace elements, PAHs, brominated flame retardants and mycotoxins. The study design, sampling methodology and primary conclusions of the study are given. It was found that in some cases dioxin-like compounds were present at levels that are of concern for the health of the egg consumers. Therefore, measures to limit their contamination in eggs, produced by hens of private owners, were proposed and deserve further attention.

Ochratoxin A in domestic and imported beers in Belgium: occurrence and exposure assessment.

Determination of ochratoxin A (OTA) concentration was performed in commercial beer in Belgium using immunoaffinity column (OchraTest™) clean-up and liquid chromatography. The procedure was validated and fulfilled the European Committee for Standardizations criteria. It offered a detection limit of 3 ng l-1 and a quantification limit of 10 ng l-1. Recovery experiments carried out with the spiked samples in the range 50–200 ng OTAl-1 showed an overall average recovery rate of 97% (RSD = 2.8%). The validated method was applied to the analysis of 62 Belgian beers and 20 commercial beers imported from Denmark, France, Germany, Ireland, Mexico, The Netherlands and Scotland. None of these beers exceeded the previously suggested EU limit of 200 ng l-1. However, OTA was detected in 60 Belgian beers and in all imported beers. The average levels of contamination were 33 ng l-1 (RSD = 112%) and 32 ng l-1 (RSD = 81%), respectively. The highest level found was 185 ng l-1. On the basis of the established tolerable daily intake (TDI) of 5 ng kg-1 body weight, accepted by the scientific committee on food of the EU, this study indicates that beer consumption in Belgium is not likely to contribute to more than a few per cent of the TDI based on the average consumption. This study also shows variability of the OTA contamination in beer with time. Thus, there is a potential risk of having highly contaminated batches from time to time. We therefore recommend to control further the OTA contamination in brewery products and to take precautionary measures during harvest, transport and storage of the raw materials to maintain the OTA intake at the lowest achievable level.

Development and application of analytical methods for the determination of mycotoxins in organic and conventional wheat

The aim of this study was to develop a multicomponent analytical method for the determination of deoxynivalenol (DON), ochratoxin A (OTA) and zearalenone (ZEN), nivalenol (NIV), 3-acetyl-DON (3-acDON), 15-acetyl-DON (15-acDON), zearalenol (ZOL) and citrinin (CIT) in wheat. It also aimed to survey the presence and amounts of DON, OTA and ZEN in Belgian conventionally and organically produced wheat grain and in wholemeal wheat flours. After solvent extraction, an anion-exchange column (SAX) was used to fix the acidic mycotoxins (OTA, CIT), whilst the neutral mycotoxins flowing through the SAX column were further purified by filtration on a MycoSep cartridge. OTA and CIT were then analysed by high-performance liquid chromatography (HPLC) using an isocratic flow and fluorescence detection, while the neutral mycotoxins were separated by a linear gradient and detected by double-mode (ultraviolet light fluorescence) detection. The average DON, ZEN and OTA recovery rates from spiked blank wheat flour were 92, 83 and 73% (RSDRu2009=u200912, 10 and 9%), respectively. Moreover, this method offered the respective detection limits of 50, 1.5 and 0.05u2009µgu2009kg−1 and good agreement with reference methods and inter-laboratory comparison exercises. Organic and conventional wheat samples harvested in 2002 and 2003 in Belgium were analysed for DON, OTA and ZEN, while wholemeal wheat flour samples were taken from Belgian retail shops and analysed for OTA and DON. Conventional wheat tended to be more frequently contaminated with DON and ZEN than organic samples, the difference being more significant for ZEN in samples harvested in 2002. The mean OTA, DON and ZEA concentrations were 0.067, 675 and 75u2009µgu2009kg−1 in conventional samples against 0.063, 285 and 19u2009µgu2009kg−1 in organically produced wheat in 2002, respectively. Wheat samples collected in 2003 were less affected by DON and ZEN than the 2002 harvest. Organic wholemeal wheat flours were more frequently contaminated by OTA than conventional samples (pu2009<u20090.10). The opposite pattern was shown for DON, organic samples being more frequently contaminated than conventional flours (pu2009<u20090.10).

Ochratoxin A and citrinin loads in stored wheat grains: Impact of grain dust and possible prediction using ergosterol measurement

Abstract Crop storage should be carried out under hygienic conditions to ensure safe products, but sometimes grain dust which has settled from previous storage may be left over and incorporated to the following stored grains. This paper describes the results obtained using a lab model developed in order to assess the impact of grain dust incorporation for its direct contribution as a contaminant but also as an inoculum in stored wheat. Settled grain dust (4 samples) released from Belgian grain storages were collected and analysed by HPLC for ergosterol, ochratoxin A (OTA) and citrinin (CIT) content. For OTA and for ergosterol, there was a high degree of variability in concentrations found in the dust samples (from 17.3–318u2009ngu2009g−1 and from 39–823u2009µgu2009g−1, respectively) whilst for CIT, the range was less significant (from 137–344u2009ngu2009g−1). Incorporation of grain dust into wheat storage contributed to an increase in the concentrations of mycotoxins in the stored grain. Dust acts as a contaminant and as an inoculum. According to these two ways, patterns of mycotoxin generation vary with the nature of the mycotoxin, the mycotoxigenic potential of dust and the water activity of the wheat. OTA and CIT showed a very versatile image when considering the amounts of toxins produced under the selected experimental conditions. The development of a robust tool to forecast the mycotoxigenicity of dust was based on the determination of ergosterol content as a general marker of fungal biomass. Present results suggest that this predictive tool would only be valid for predicting the contamination level of CIT and OTA at reasonable moisture content (14–20%). The potential risk of having highly contaminated batches from stock to stock may thus occur and this paper discusses possible pathways leading to OTA and CIT contamination either under wet or dry storage conditions. We therefore, recommend taking precautionary measures not only by controlling and maintaining moisture at a reasonable level during storage of the raw materials but also by paying more attention to the cleaning of the stores before loading in the new harvests.

Transfer of ochratoxin A during lactation: exposure of suckling via the milk of rabbit does fed a naturally-contaminated feed.

The transfer of ochratoxin A from the blood to the milk of lactating rabbit does and subsequently the exposure of their sucklings to the mycotoxin were investigated. An effective transfer of ochratoxin A from blood to milk was shown in lactating rabbit does fed a naturally-contaminated diet (10-20g/kg of body weight/day) throughout a lactation period of 19 days. The ochra-toxin A concentrations in plasma and in milk did not significantly change throughout the lactation period with a mean milk/plasma concentration ratio of 0.015. These variables were however significantly correlated (p < 0.05), as were the ingested amounts and milk concentrations. At slaughter, the highest concentration of ochratoxin A accumulated in the body of the rabbit does were found in kidney (1.2 g/kg) followed by liver (158ng/kg), mammary gland (105ng/kg) and muscle (38 ng/kg). A linear relationship was found between the ochratoxin A concentrations in milk and in the plasma of the sucklings, indicating an effective transfer of the toxin to the sucklings. If the same is true in humans, the exposure of the breast-fed infant to the toxin, which has been largely reported in the literature, should be a major matter of concern for human health.

Mycotoxin analyses in some home produced eggs in Belgium reveal small contribution to the total daily intake

Low levels of deoxynivalenol (DON, range: 2.6-17.9 ng/g) and its metabolite de-epoxy-DON (DOM-1, range: 2.4-23.7 ng/g) were found in 20 home-produced egg samples collected in Belgium during autumn 2006 (from 10 breeders) and spring 2007 (same breeders). DON intake assessment showed that the consumption of these eggs may contribute to less than 1% of the provisional maximum tolerable daily intake of 1 microg/kg body weight established by FAO/WHO. None of the egg samples analyzed had quantifiable levels of zearalenone (ZEA), alpha-zearalenol, beta-zearalenol, ochratoxin A (OTA) and citrinin (CIT). Intake of DON, ZEA, OTA and CIT via the consumption of home produced eggs seems not to be a matter of concern. Despite this, we recommend to continue in screening other eggs allowing to increase the sample size and the subsequent conclusion for mycotoxin contamination in eggs. As home produced food is generally not submitted to any compliance control and may be consumed in large quantities by their producers and other household members, it is worthwhile to further pay attention to the quality of feed as well as the environment in which the hens live.

Cross-reactivity of some commercially available deoxynivalenol (DON) and zearalenone (ZEN) immunoaffinity columns to DON- and ZEN-conjugated forms and metabolites

Seven commercially available deoxynivalenol (DON) and zearalenone (ZEN) immunoaffinity columns (IACs) were tested for cross-reactivity to conjugated forms (3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, DON-3-glucoside, DON-3-glucuronide, ZEN-glucosides, ZEN-glucuronide) and metabolites (de-epoxydeoxynivalenol, α-zearalenol, β-zearalenol) and nivalenol (NIV), using a semi-quantitative multi-mycotoxin ultra-performance liquid chromatography-tandem mass spectrometry method. The DON IACs showed cross-reactivity for nearly all DON derivatives tested. The ZEN IACs showed limited cross-reactivity to some of the ZEN derivatives. The IACs were evaluated for their potential use as sample clean-up for mycotoxins in serum.

Intake of ochratoxin A and deoxynivalenol through beer consumption in Belgium.

Estimations of ochratoxin A (OTA) and 4-deoxynivalenol (DON) exposure of the Belgian population through beer consumption were made using the results of the recent Belgian food survey and the compiled data set of OTA and DON levels in conventionally and organically produced beers in 2003–05. For the consumers of organic beers, the daily intake of OTA was 0.86 (in 2003), 1.76 (in 2004) and 0.72 (in 2005) ngu2009kg−1 body weight (bw), considering the mean beer consumption (0.638 litres) and the average level of OTA in 2003, 2004 and 2005, respectively. Using the 97.5th percentile of beer consumption (1.972 litres), the corresponding OTA daily intakes were 2.65, 5.44 and 2.24u2009ngu2009kg−1u2009bw, which are close or above the tolerable daily intake (TDI) of 5u2009ngu2009kg−1u2009bw. For the consumers of conventional beers, the OTA intakes were low: 0.23, 0.23 and 0.11u2009ngu2009kg−1u2009bwu2009day−1 for the average beer consumption, in 2003, 2004 and 2005 against 0.72, 0.73 and 0.34u2009ngu2009kg−1u2009bwu2009day−1 when the 97.5th percentile level was considered. As for the DON intake, the estimates were quite low for both conventional and organic beer consumers when the provisional maximum TDI (PMTDI) of 1u2009µgu2009kg−1 bw was considered. Average consumption of organic beer led to daily intakes of 0.05 and 0.04u2009µgu2009DONu2009kg−1u2009bw in 2003 and 2004, respectively, whilst for conventional beer, daily intakes were 0.07 and 0.05u2009µgu2009DONu2009kg−1u2009bw. At the 97.5th percentile level of beer consumption, daily intakes of 0.15 and 0.13u2009µgu2009kg−1u2009bw were obtained for organic beers against 0.23 and 0.17u2009µgu2009kg−1u2009bw for conventional ones. The results showed that beer could be an important contributor to OTA exposure in Belgium, even though a declining trend seems to be apparent during the last year of monitoring. Therefore, efforts should be devoted to maintain the OTA levels as low as reasonably achievable, especially for organic beer.