Deoxynivalenol and other Fusarium mycotoxins in bread,cake, and biscuits produced from UK-grown wheat under commercial and pilot scale conditions

Bread, cakes and biscuits were manufactured from flour containing deoxynivalenol (DON) and low concentrations of zearalenone (ZON) and nivalenol (NIV). The results show that these mycotoxins remain mostly unaffected during manufacture. Although the results indicate that the mycotoxins are largely stable and survive processing, when concentrations were determined on an ‘as is’ basis as stipulated in legislation, levels in finished products were usually lower than in the starting flour due to the dilution effect of other ingredients such as fat, sugar and water. Thus mean concentrations of DON in bread were reduced by about 35% and 39% in white and wholemeal bread respectively which are in close agreement with the reduction required by the regulations although the changes that occur during milling white and wholemeal flour from whole wheat also need to be taken into account. The reduction of DON during cake manufacture is greater than for bread because flour makes up only about 25% of the starting ingredients. However, in biscuit production, particularly for crackers for which flour constitutes about 90% of the recipe ingredients, the reduction indicated by the regulations is not achieved. It is concluded that for some commercial processes, the whole-wheat or flour ingredients used will need to contain DON levels lower than those set by legislation to ensure that the final products will still meet statutory limits. Limited results with consignments containing low concentrations of ZON and NIV suggest that their stability and survival are similar to those for DON.     

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% (RSDR = 12, 10 and 9%), respectively. Moreover, this method offered the respective detection limits of 50, 1.5 and 0.05 microg kg-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 75 microg kg-1 in conventional samples against 0.063, 285 and 19 microg kg-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 (p < 0.10). The opposite pattern was shown for DON, organic samples being more frequently contaminated than conventional flours (p < 0.10).     

Analysis of trichothecenes in laboratory rat feed by gas chromatography-tandem mass spectrometry

A method for the determination of seven trichothecenes, neosolaniol (NEO), diacetoxyscirpenol (DAS), deoxynivalenol (DON), nivalenol (NIV), fusarenon-X (FUS-X), 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON), in laboratory rat feed by GC-MS/MS was developed. Sample extraction and purification was performed by an acidified mixture of acetonitrile/water (80–20% v/v). Limits of quantitation (LOQs) were between 1 and 10 μg kg^–1 for all studied trichothecenes. Eight concentration levels between the LOQ and 100 × LOQ were used for the calibration curves. Matrix-matched calibration was used for quantitation purposes to compensate the detector signal enhancement obtained for all the analytes. The method accuracy was evaluated by recovery assays at three concentration levels, 25, 50 and 100 μg kg^–1 (n = 9). Recoveries ranged from 62% to 97% and precision, expressed as intra- and inter-day relative standard deviations, was evaluated for all compounds. The validated method was successfully applied to the analysis of 35 laboratory rat feed samples showing mycotoxin contamination in 66% of the samples. DON was the most prevalent trichothecene followed by 15-ADON, NIV and 3-ADON. The maximum DON concentration reached in real samples was 2156 ± 4.3 μg kg^–1, while NEO, DAS and FUS-X were not detected in any sample. Multi-contamination by at least two mycotoxins was observed in 17% of the analysed feed samples.     

Molecular survey of trichothecene genotypes of Fusarium graminearum species complex from barley in southern Brazil

Fusarium head blight is a disease of primary concern to small-grain cereals of Brazil, including barley. Its main causal agent, Fusarium graminearum species complex (Fg complex)¸ is able to produce mycotoxins, especially deoxynivalenol (DON) and nivalenol (NIV), that usually contaminate grain. Strains that produce DON may also produce its acetylated derivatives: 3-acetyl-DON (3-ADON) and 15-acetyl-DON (15-ADON). Ninety two isolates were obtained from samplings of barley grain during three years (2007, 2008 and 2009) from several fields in both southern and northern production regions of Rio Grande do Sul state, Brazil. These isolates were examined for polymerase chain-reaction-based (PCR) trichothecene genotype based on the amplification of portions of Tri3 and Tri12. There was no effect of year or region on the proportion of trichothecene genotypes. Overall, 66% of the strains (61/92) were 15-ADON, 4.4% (4/92) were 3-ADON and 29.3% (27/92) were NIV. The overall NIV/DON ratio estimated (0.41) was five times higher than that found in previous studies with strains from wheat grown in the same region. Species identification of nine strains representing the trichothecene genotypes, based on comparisons of DNA sequences of portions of the PHO, RED and URA genes with sequences from curated reference isolates of Fusarium from GenBank, revealed that they belong to F. graminearum sensu stricto (four 15-ADON and one 3-ADON strain), F. meridionale (three NIV strains) and F. austroamericanum (one 3-ADON strain). These results add to the current regional knowledge of trichothecene genotypes and species within the Fg complex affecting barley in the region.     

Genotyping and phenotyping of Fusarium graminearum isolates from Germany related to their mycotoxin biosynthesis

Fusarium graminearum is the most important pathogen causing Fusarium head blight (FHB) of small cereal grains worldwide responsible for quantitative and qualitative yield losses. The presence in crops is often associated with mycotoxin contamination of foodstuff limiting its use for human and animal consumption. A collection of isolates of F. graminearum from Germany was characterized genetically and chemically for their potential to produce the B trichothecenes deoxynivalenol (DON) and nivalenol (NIV). Molecular methods with eight PCR assays were implemented based on functional Tri7 and Tri13 genes and on the tri5-tri6 intergenic region to differentiate between chemotaxonomic groups DON and NIV, resulting in a marked majority (61/63) of DON chemotypes. Mycotoxins produced on rice kernels were quantified by means of LC-MSMS including DON, NIV, 3-acetyl-DON (3-ADON), 15-acetyl-DON (15-ADON), DON-3-glucoside, fusarenon X, as well as zearalenone; all of them proving to be present in high concentration among the isolates. All DON-chemotype isolates also produced lower amounts of NIV with the amount being positively correlated (R² = 0.89) to the DON amount. 15-ADON and 3-ADON are reported to be produced simultaneously by the isolates, the former dominating over the latter in all but one isolate. Fungal biomass, was quantified via ergosterol amount on rice. It was used to calculate specific mycotoxin production per biomass of isolates, ranging from 0.104 to 1.815 mg DON mg-1 ergosterol, presenting a Gaussian distribution. Genotype and phenotype characterization revealed discrepancies with respect to mycotoxin production potential of the fungi, i.e. isolates from one chemotype were able to produce mycotoxins from other chemotypes in considerable amounts.     

Effect of the bread-making process on zearalenone levels

The effects of the bread-making process including fermentation with Saccharomyces cerevisiae and lactic acid bacteria (Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus and Lactobacillus fermentum) and baking at 200°C on zearalenone (ZEA) levels were investigated. Standard solutions of ZEA were added to flour and then loaves of bread were prepared. Sourdough and three types of yeast including active dry yeast, instant dry yeast and compressed yeast were used for the fermentation of dough. ZEA levels in flour, dough and bread were determined by HPLC with fluorescence detection after extraction and clean-up on an immunoaffinity column. The highest reduction in levels of ZEA was found in the first fermentation (first proof), while the lowest reduction was observed in the baking stage. In addition, the results showed that compressed yeast had the maximum reduction potential on ZEA levels even at the baking stage.     

Natural deoxynivalenol occurrence and genotype and chemotype determination of a field population of the Fusarium graminearum complex associated with soybean in Argentina

Soybean (Glycine max L.), the main source of protein throughout the world, is used both as a food and a feedstuff. Currently, limited information about the occurrence of Fusarium species and mycotoxins in soybean grain and by-products is available. The aims of the present study were: (1) to identify toxigenic Fusarium species associated with soybean during crop reproductive stages; (2) to determine the occurrence of deoxynivalenol (DON) and nivalenol (NIV) in soybean seeds; (3) to determine the genotype and chemotype of selected Fg complex strains using molecular and chemical analysis, respectively; and (4) to characterize the strains using AFLP(s) markers. One soybean field located at Córdoba Province, Argentina, was monitored and samples of soybean tissue were harvested at three reproductive stages: flowering (R2), full seed (R6) and full maturity (R8). A total of 389 Fusarium strains F. equiseti (40%) was the most frequently species recovered followed by F. semitectum (27%) and F. graminearum (Fg) (11%). From the 40 soybean samples analysed, only two presented detectable DON levels. Based on DON occurrence on soybean seeds at ripening stages, the toxigenic ability of Fg complex strains isolated from soybean seeds, pods and flowers were analysed. The trichothecene genotype was determined by a multiplex PCR using primers based on Tri3, Tri5 and Tri7 toxin genes and then the chemotype was verified by chemical analysis. Most Fg complex strains showed 15-ADON genotype and five strains presented a DON/NIV; these also produced both toxins under in vitro culture. Neither the NIV nor the 3-ADON genotypes were detected among the members of the population evaluated. All the 15-ADON genotype strains were characterized as F. graminearum sensu stricto (lineage 7), while the strains presented a DON/NIV genotype were characterized as F. meridionale (lineage 2). The present study contributes new information on the occurrence of Fusarium species and trichothecenes toxins on soybean at the pre-harvest stages. Also, this is the first report on the chemotype, genotype and lineages among Fg complex isolated from soybean.     

Natural deoxynivalenol occurrence and genotype and chemotype determination of a field population of the Fusarium graminearum complex associated with soybean in Argentina

Soybean (Glycine max L.), the main source of protein throughout the world, is used both as a food and a feedstuff. Currently, limited information about the occurrence of Fusarium species and mycotoxins in soybean grain and by-products is available. The aims of the present study were: (1) to identify toxigenic Fusarium species associated with soybean during crop reproductive stages; (2) to determine the occurrence of deoxynivalenol (DON) and nivalenol (NIV) in soybean seeds; (3) to determine the genotype and chemotype of selected Fg complex strains using molecular and chemical analysis, respectively; and (4) to characterize the strains using AFLP_s markers. One soybean field located at Córdoba Province, Argentina, was monitored and samples of soybean tissue were harvested at three reproductive stages: flowering (R2), full seed (R6) and full maturity (R8). A total of 389 Fusarium strains F. equiseti (40%) was the most frequently species recovered followed by F. semitectum (27%) and F. graminearum (Fg) (11%). From the 40 soybean samples analysed, only two presented detectable DON levels. Based on DON occurrence on soybean seeds at ripening stages, the toxigenic ability of Fg complex strains isolated from soybean seeds, pods and flowers were analysed. The trichothecene genotype was determined by a multiplex PCR using primers based on Tri3, Tri5 and Tri7 toxin genes and then the chemotype was verified by chemical analysis. Most Fg complex strains showed 15-ADON genotype and five strains presented a DON/NIV; these also produced both toxins under in vitro culture. Neither the NIV nor the 3-ADON genotypes were detected among the members of the population evaluated. All the 15-ADON genotype strains were characterized as F. graminearum sensu stricto (lineage 7), while the strains presented a DON/NIV genotype were characterized as F. meridionale (lineage 2). The present study contributes new information on the occurrence of Fusarium species and trichothecenes toxins on soybean at the pre-harvest stages. Also, this is the first report on the chemotype, genotype and lineages among Fg complex isolated from soybean.     

Trichothecene genotypes and chemotypes in Fusarium graminearum strains isolated from wheat in Argentina

Argentina is the fourth largest exporter of wheat in the world. The main pathogen associated with Fusarium Head Blight (FHB) of wheat in Argentina is Fusarium graminearum lineage 7 also termed F. graminearum sensu stricto in the F. graminearum species complex, which can produce the Type B trichothecenes, usually deoxynivalenol (DON) and its acetylated forms (3-ADON and 15-ADON) or nivalenol (NIV). We used a multiplex PCR assay of Tri3, Tri7, and Tri13 to determine the trichothecene genotype of 116 strains F. graminearum collected from three locations in Argentina and then verified the chemotype by chemical analysis. PCR assays and chemical analyses gave the same results for all strains that produced trichothecenes. Most strains (> 92%) had the 15-ADON genotype, with the remaining strains having the DON/NIV genotype. We observed neither the NIV nor the 3-ADON genotypes amongst the strains evaluated. The nine strains with the DON/NIV genotype produced DON when analyzed chemically. Thus, the Argentinean populations of F. graminearum are similar to those from wheat elsewhere in the world, in that all the strains produced DON/15-ADON and belong to lineage 7. However approximately 8% of the strains tested were incorrectly diagnosed as DON/NIV producers with the current multiplex PCR and were only DON producers by chemical analysis.     

Effect of fermentation on naturally occurring deoxynivalenol (DON) in Argentinean bread processing technology.

The stability of naturally occurring DON was evaluated during the fermentation stage of the bread-making process on a pilot scale. Two different products, French bread and Vienna bread, were prepared with naturally contaminated wheat flour (150 mg kg(-1)) under controlled experimental conditions. Dough was fermented at 30, 40 and 50 degrees C according to standard procedures employed in Argentinean low-technology bakeries. When the dough was fermented at 50 degrees C, the maximum reduction was 56% for the Vienna bread, with French bread being reduced by 41%. DON reduction during bread-making occurs not only in the baker due to thermal decomposition, but also during the fermentation step. The Argentinean traditional bread-making process might reduce DON levels during the fermentation stages if the dough is leavened at temperatures > 30 degrees C.     

Fusarium toxins in wheat from an area in Henan Province, PR China, with a previous human red mould intoxication episode.

Wheat samples of the 1998 and 1999 crops from Puyang, an area in Henan Province, PR China with a previous human red mould intoxication episode, were analysed for trichothecenes and zearalenone (ZEA). For the 1998 Puyang crop, deoxynivalenol (DON) was the predominant toxin detected abundantly and frequently at a level of up to 14,000 microg kg(-1) (mean 2850 microgkg(-1)) in 30 of 31 (97%) wheat samples. Among these were 21 (70%) with a DON level that exceeded the Chinese regulation of 1,000 microg kg(-1). Nivalenol (NIV) and 15-acetyl-DON (15-ADON) were also found at 578 microg kg(-1) (one sample) and 59-1,800 microg kg(-1) (mean 365 microg kg(-1), 20 samples), respectively. ZEA co-occurred in 21 samples at 9-1,400 microg kg(-1) (mean 209 microg kg(-1)). Twenty-five (89%) wheat samples from Zhumadian, a region without a history of human red mould intoxication in the same province, contained low levels of DON (53-1240, mean 223 microg kg(-1)) with seven (25%) co-contaminated with ZEA (10-217, mean 108 microg kg(-1)). All were free from 15-ADON and NIV. Significant differences in DON, 15-ADON and ZEA concentrations between both areas were found. DON (<1000 microg kg(-1)) and ZEA (5-111 microg kg(-1)) were also detected in the 1999 Puyang wheat. Proper environmental conditions for Fusarium species surviving winter combined with unusual high precipitation during wheat flowering were responsible for a high concentration of Fusarium mycotoxins in the 1998 Puyang wheat.     

真菌毒素在赤霉病小麦研磨及其湿法加工中的分布研究

小麦赤霉病不仅会导致粮食减产,更会引起多种真菌毒素的高污染风险。将染病小麦进行实验室制粉并湿法分离统粉中的粗淀粉、谷朊粉,采用酶联免疫吸附测定法(Enzyme-Linked Immuno Sorbent Assay,ELISA)测定各系统的呕吐毒素(Deoxynivalenol,DON)、黄曲霉毒素B1(Aflatoxin B1,AFB1)、玉米赤霉烯酮(Zearalenone,ZEN)以及赭曲霉毒素A(Ochratoxin A,OTA)的含量,以探究赤霉病小麦在制粉及其湿法加工中四种常见真菌毒素的分布变化规律。结果显示:研磨制粉及湿法加工对真菌毒素的分布影响显著。制粉加工后,皮磨和心磨系统粉的DON、AFB1、ZEN和OTA含量比原粮显著降低,降低率分别为1.38~16.24%、20.47~71.77%及26.71~69.51%;湿法加工产物中,DON消减为相应统粉的4.88~12.11%,AFB1与OTA浓缩富集,富集率分别可达统粉的2.55、3.65倍,粗淀粉中ZEN消减为统粉的12.70~15.83%,谷朊粉则富集为统粉的4.11倍。研究表明,在工业生产中,可根据赤霉病小麦的感染类型及程度,适当选用研磨或湿法加工等合适的方法加工处理。     

Effect of sourdough processing and baking on the content of enniatins and beauvericin in wheat and rye bread

The occurrence of the emerging mycotoxins enniatins (ENNs) and beauvericin (BEA) has been reported in Fusarium-infected cereals. To study the effect of sourdough processing and baking on ENN B, ENN B1, and BEA concentrations, a recently developed stable isotope dilution assay for these mycotoxins was used. After milling of wheat and rye grains naturally contaminated with ENN B and ENN B1, approximately 70–82 % of the two ENNs were found in the bran fraction and the rest remained in flour. BEA was added to flour before sourdough fermentation. In an experiment on a microscale, dough was fermented for 24 h at 30 or 40 °C, which reduced part of the ENNs and BEA in particular at 40 °C. On a standard scale, mixing, resting, and proofing of the bread dough resulted in 13–19 % reduction of the ENNs compared with flour, but in no significant change of BEA. The final baking at 200 °C for 25 min led to a further decrease of the ENNs and BEA, ranging from 9 to 28 % compared with fermented dough. In case of rye sourdough bread, greater reductions of ENNs were found in crust than in crumb. For both wheat and rye flours, overall 25–41 % of ENN B, ENN B1, and BEA were reduced during the whole sourdough bread-making process.     

Incidence of trichothecenes and zearalenone in poultry feed mixtures from Slovakia

A total of 50 samples of poultry feed mixtures of Slovakian origin were analyzed for eight toxicologically significant Fusarium mycotoxins, namely zearalenone (ZON), A-trichothecenes: diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2) and B-trichothecenes: deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), 15-acetyl-deoxynivalenol (15-ADON) and nivalenol (NIV). The A-trichothecenes and the B-trichothecenes were detected by means of high pressure liquid chromatography with tandem mass spectrometry detection (HPLC-MS/MS) and gas chromatography electron capture detection (GC-ECD), respectively. Reversed phase-high performance liquid chromatography with a fluorescence detector (RP-HPLC-FLD) was used for ZON detection. The most frequent mycotoxin detected was T-2, which was found in 45 samples (90%) in relatively low concentrations ranging from 1 to 130 microg kg(-1) (average 13 microg kg(-1)), followed by ZON that was found in 44 samples (88%) in concentrations ranging from 3 to 86 microg kg(-1) (average 21 microg kg(-1)). HT-2 and DON were detected in 38 (76%) and 28 (56%) samples, respectively, in concentrations of 2 to 173 (average 18 microg kg(-1)) for HT-2 and 64 to 1230 microg kg(-1) sample (average 303 microg kg(-1)) for DON. The acetyl-derivatives of DON were in just four samples, while NIV was not detected in any of the samples investigated. In as many as 22 samples (44%), a combination of four simultaneously co-occurring mycotoxins, i.e. T-2, HT-2, ZON and DON, was revealed. Despite the limited number of samples investigated during this study poultry feed mixtures may represent a risk from a toxicological point of view and should be regarded as a potential source of the Fusarium mycotoxins in Central Europe. This is the first reported study dealing with zearalenone and trichothecene contamination of poultry mixed feeds from Slovakia.     

The prevalence of deoxynivalenol and its derivatives in the spring wheat grain from different agricultural production systems in Lithuania

Deoxynivalenol (DON) together with two acetylated derivatives, 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON) occurs in cereal grains and their products. Co-occurrence of DON and acetylated derivatives in cereal grain is detected worldwide. Until now, DON and its derivatives have been considered equally toxic by health authorities. In this study, we analysed 103 samples of spring wheat grain, originating from the fields of different production systems in Lithuania, for the co-occurrence of type-B trichothecenes (DON, 3-ADON, 15-ADON). The samples were classified according to the production system—organic, sustainable and intensive. Mycotoxin levels in the spring wheat grain samples were determined by the HPLC method with UV detection. The type-B trichothecenes were found to be present at higher concentrations in the grain from the intensive production system. Eighty-one percent of the spring wheat grain samples from the intensive production system were co-contaminated with a combination of DON+3-ADON+15-ADON, 1% with DON+3-ADON. Additionally, DON+15-ADON and DON were found in 5% and 10% of the tested samples, respectively. Two percent of the samples were free from mycotoxins. In the grain samples from the sustainable production system, DON and a combination of DON+3-ADON showed a higher incidence – 47% and 23%, respectively. The samples with a combination of DON+3-ADON+15-ADON accounted for 18%. Completely different results were obtained from the analyses of organic grain samples. A large number of the organic spring wheat grain samples were contaminated with DON+3-ADON (55%) or DON (36%). The combination of DON+3-ADON+15-ADON was not present, while DON+15-ADON was present in 9% of the samples tested. The production systems did not lead to significant differences in mycotoxin levels, although a trend toward higher incidence and higher contamination was observed for the samples from the intensive and sustainable production systems.     

Fate of deoxynivalenol and nivalenol during storage of organic whole-grain wheat flour

This study was carried out to determine the level of retention of the mycotoxins deoxynivalenol (DON) and nivalenol (NIV) during 120 days of storage (aging) of flours produced from organic wheat grain naturally infected with Fusarium fungi. Three types of flour (standard white flour prepared by a roller-grinder mill - IRG, whole-grain flour produced by a hammer-crusher mill - IHC and whole-grain flour prepared by a millstone - OMS) were packaged in food-grade paper or polypropylene plastic bags and stored at two different storage temperatures (constant 10 °C or 25 °C). The concentrations of DON and NIV were measured prior to and after storage by means of HPLC-UV detection methods. After 120 days of storage, the concentrations of DON and NIV decreased between 0% and 29% compared to the initial measurements, depending on the combination of experimental factors. The greatest decrease in mycotoxin concentration was observed in the IHC and OMS flours packaged in paper bags and stored at 25 °C. The smallest decrease in mycotoxin concentration was observed in the IRG flours packaged in sealed plastic bags and stored at 10 °C. Statistical analysis showed that the level of retention of DON and NIV depended significantly on the type of packaging material, but did not depend on the type of flour or the storage temperature.     

Relationship between Fusarium spp. diversity and mycotoxin contents of mature grains in southern Belgium

Over a 4-year period (2010–13), a survey aiming at determining the occurrence of Fusarium spp. and their relations to mycotoxins in mature grains took place in southern Belgium. The most prevalent species were F. graminearum, F. avenaceum, F. poae and F. culmorum, with large variations between years and locations. An even proportion of mating type found for F. avenaceum, F. culmorum, F. cerealis and F. tricinctum is usually a sign of ongoing sexual recombination. In contrast, an unbalanced proportion of mating type was found for F. poae and no MAT1-2 allele was present in the F. langsethiae population. Genetic chemotyping indicates a majority of deoxynivalenol (DON)-producing strains in F. culmorum (78%, all 3-ADON producers) and F. graminearum (95%, mostly 15-ADON producers), while all F. cerealis strains belong to the nivalenol (NIV) chemotype. Between 2011 and 2013, DON, NIV, enniatins (ENNs) and moniliformin (MON) were found in each field in various concentrations. By comparison, beauvericin (BEA) was scarcely detected and T-2 toxin, zearalenone and α- and β-zearalenols were never detected. Principal component analysis revealed correlations of DON with F. graminearum, ENNs and MON with F. avenaceum and NIV with F. culmorum, F. cerealis and F. poae. BEA was associated with the presence of F. tricinctum and, to a lesser extent, with the presence of F. poae. The use of genetic chemotype data revealed that DON concentrations were mostly influenced by DON-producing strains of F. graminearum and F. culmorum, whereas the concentrations of NIV were influenced by the number of NIV-producing strains of both species added to the number of F. cerealis and F. poae strains. This study emphasises the need to pay attention to less-studied Fusarium spp. for future Fusarium head blight management strategies, as they commonly co-occur in the field and are associated with a broad spectrum of mycotoxins.     

Effect of food processing on exposure assessment studies with mycotoxins

The goals of the present work were, on the one hand, to assess the effect of baking on the stability of zearalenone (ZEA) and deoxynivalenol (DON), as well as the transfer of DON from pasta to boiling water, and, on the other hand, to quantify the impact of DON depletion, during cooking of pasta, on overall exposure estimates. Therefore, the bread-making process was simulated on a pilot-plant scale by using naturally contaminated flour with DON and ZEA. Transfer of DON from pasta to water was evaluated at different boiling times. Pasta was prepared on a pilot-plant scale by using naturally contaminated durum wheat flour; subsequently, it was boiled simulating home cooking. The experiments examined the stability of DON and ZEA during the bread-making process, including fermentation with Saccharomyces cerevisiae and baking at 200°C. Our results showed a high transfer of DON from pasta to boiling water, reaching depletion levels of almost 75%, which correlated with levels found in water. Accordingly, these cooking depletion rates were computed through a stochastic exposure model to weight their impact on the final exposure estimates. Finally, statistically significant differences were found in most of the parameters and populations assessed, but these were not enough to consider the process as protective because the contribution of pasta to the overall DON intake was commonly low.     

Natural occurrence of Fusarium toxins in oats harvested during five years in an area of southwest Germany.

A total of 56, 56, 54, 51, and 55 oats samples used for feed production were collected randomly after the 1987, 1989, 1990, 1991 and 1992 crops, respectively, from farms located in an area of southwest Germany. Deoxynivalenol (DON), 3- and 15-acetyl-deoxynivalenol (3-, 15-ADON), nivalenol (NIV), fusarenon-X (FUS-X), T-2 toxin (T-2), HT-2 toxin (HT-2) and diacetoxyscirpenol (DAS) were determined by gas chromatography with mass selective detection (GC-MS), zearalenone (ZEA), alpha and beta-zearalenol (alpha-, beta-ZOL) by GC-MS or by HPLC. DON was the major toxin with incidences at 49-85% and mean levels in positive samples of 52-302 micrograms/kg. Incidences of ZEA, 3-ADON, NIV, HT-2, and T-2 were at 20-37, 0-30, 18-67, 0-29, and 27-61%, respectively, with mean levels in positive samples at 8-25, 5-63, 11-192, 205-296, and 20-244 micrograms/kg, respectively. alpha- and beta-ZOL and DAS were not detected in any sample. 15-ADON and FUS-X were assayed in samples from 1987, 1991 and 1992. 15-ADON was detected in 9, 4 and 0% of samples, with an average of 9 and 18 micrograms/kg, respectively; FUS-X was not detected. The incidence and levels of toxins varied from year to year. The correlation between the occurrence of toxins and precipitation is discussed.