Effect of gamma-irradiation on the natural occurrence of Fusarium mycotoxins in wheat,flour and bread

A survey was carried out to obtain data on the occurrence of Fusarium mycotoxin in wheat and flour samples collected from local markets in Egypt and to study the influence of gamma-irradiation on controlling the occurrence of these mycotoxins in wheat, flour and bread. Deoxynivalenol (DON) was detected in five samples of wheat at levels ranging from 103 to 287 μg/kg and one sample each of flour and bread at concentrations 188 and 170 μg/kg. Zearalenone (ZEN) was detected in ten samples of wheat at levels from 28 to 42 μg/kg and four samples each of flour and bread at concentrations of 95 and 34 μg/kg, respectively. T-2 toxin was detected only in one sample each of wheat, flour and bread at concentrations of 2.9, 2.2 and 2.3 μg/kg, respectively. Gamma-irradiation at dose level of 6 kGy completely eliminated fungal flora in flour and wheat. DON, ZEN and T-2 toxin concentrations are reduced to 85, 20 and 2.0 μg/kg for wheat and to 125, 45, and 1.0 μg/kg for flour after 4 kGy exposure and a sharp drop in Fusarium toxin levels occurred at 6 kGy and was eliminated at 8 kGy. Bread prepared from 6 kGy was contaminated with Fusarium toxin at levels below 5 μg/kg. It was noticed that gamma-irradiation reduce greatly the natural occurrence of Fusarium mycotoxins in bread.     

Changes in ochratoxin A and type B trichothecenes contained in wheat flour during dough fermentation and bread-baking

Ochratoxin A (OTA) and type B trichothecenes are mycotoxins that occur frequently in cereals and thus can be found in cereal by-products such as bread. The aim of this work was to study the variation of the levels of OTA, deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON) and nivalenol (NIV) during the bread-making process. This was done by using wheat flour spiked with different levels of toxins. Mycotoxin levels were controlled after fermentation of the dough with yeasts (Saccharomyces cerevisiae) and after further baking at different temperature–time combinations. Analysis of variance (ANOVA) of the results showed a significant reduction in OTA level (p < 0.05) during fermentation of the dough. The reduction ranged between 29.8% and 33.5%, depending on the initial concentration of toxin in the flour. During this period, the level of the other mycotoxins studied was not modified. By contrast, in the baking phase there were significant changes in the levels of the four mycotoxins, although the reduction was similar under all the baking conditions. Considering all the temperature–time conditions tested, it can be concluded that during the baking period the average reduction of OTA, NIV, 3-ADON, and DON was 32.9%, 76.9%, 65.6%, and 47.9%, respectively.     

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.     

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.     

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.     

Stability of the mycotoxin deoxynivalenol (DON) during the production of flour-based foods and wheat flake cereal

Deoxynivalenol (DON) is a mycotoxin found in cereal grains and cereal-based foods. DON concentrations in finished products are reduced under some processing conditions, but not others. DON concentrations in flour, wheat and selected foods made from them under commercially relevant conditions were compared by GC with electron capture detection. Average concentrations (n?=?9/item) in cookies, crackers and pretzels ranged from 61% (cookies) to 111% (pretzels) compared with flour (100%?=?0.46?µg?g^?1). Lesser amounts were found in donuts and bread: their respective DON concentrations were 44% and 30% that of flour. Mass balance estimates for DON (µg?g^?1 flour equivalents) ranged from 50% (bread?=?0.23?µg?g^?1 flour equivalents) to 120% (donuts), indicating that dilution by recipe ingredients contributed to DON reductions in bread and accounted for all of the apparent reduction in donuts. Mass balance estimates averaged 76% (crackers) to 107% (pretzels) for the other flour products. DON concentrations were higher in cereal flakes (0.55?µg?g^?1 in the finished product and 0.58?µg?g^?1 on a mass balance basis) than in wheat (0.40?µg?g^?1), suggesting that DON concentrations might increase during processing of wheat cereals under some conditions. In summary, DON concentrations of finished food products were reduced?≥50% only in bread and donuts. Reduction in bread resulted from a combination of DON ‘loss’ and dilution by recipe ingredients whereas the reduction in donuts was due entirely to dilution. These results are further evidence of DON stability during the preparation of popular flour or wheat-based products.     

Fusarium DNA traceability along the bread production chain

Wheat grain contamination with toxigenic Fusarium spp. is of great economic concern to cereal producers and to the grain processing industry and is of great relevance for the quality and safety of the final products. In particular, the bread production chain can potentially be a vehicle for mycotoxin ingestion above the tolerable total daily intake. A quantitative polymerase chain reaction‐based analytical approach has been developed as a possible tool to estimate and control the risk of mycotoxins, particularly deoxynivalenol (DON). This DNA‐based analytical method has been applied to detect and quantify contamination levels of Fusarium graminearum and Fusarium culmorum in naturally infected wheat grain samples. The persistence of Fusarium contamination was also detected along the bread production chain in wholemeal, flour and bread. A significant correlation was found between Fusarium DNA and DON concentration in all samples.     

Thermal degradation of deoxynivalenol during maize bread baking

The thermal degradation of deoxynivalenol (DON) was determined at isothermal baking conditions within the temperature range of 100–250°C, using a crust-like model, which was prepared with naturally contaminated maize flour. No degradation was observed at 100°C. For the temperatures of 150, 200 and 250°C, thermal degradation rate constants (k) were calculated and temperature dependence of DON degradation was observed by using Arrhenius equation. The degradation of DON obeyed Arrhenius law with a regression coefficient of 0.95. A classical bread baking operation was also performed at 250°C for 70?min and the rate of DON degradation in the bread was estimated by using the kinetic data derived from the model study. The crust and crumb temperatures recorded during bread baking were used to calculate the thermal degradation rate constants (k) and partial DON degradations at certain time intervals. Using these data, total degradation at the end of the entire baking process was predicted for both crust and crumb. This DON degradation was consistent with the experimental degradation data, confirming the accuracy of kinetic constants determined by means of the crust-like model.     

Fate of deoxynivalenol,T-2 and HT-2 toxins and their glucoside conjugates from flour to bread: an investigation by high-performance liquid chromatography high-resolution mass spectrometry

Deoxynivalenol, T-2 and HT-2 toxins are mycotoxins frequently occurring in cereals and cereal-based products along with their conjugated forms. In this paper, we provide insights into the fate of deoxynivalenol, T-2 and HT-2 toxins and their glucoside derivatives during bread making, using naturally contaminated wheat flour. High-resolution mass spectrometry was used to assess the extent of degradation of the three mycotoxins during bread baking and to identify some glucoside conjugates, namely deoxynivalenol, T-2 and HT-2 mono-glucosides, detected both in the flour and in the respective breads. Our findings show deoxynivalenol's levels markedly increased upon baking, whereas those of HT-2 and T-2 toxins were decreased in the final bread with special regard to the T-2 toxin.     

Fate of Fusarium mycotoxins in maize flour and grits during extrusion cooking

Extrusion technology is used widely in the manufacture of a range of breakfast cereals and snacks for human consumption and animal feeds. To minimise consumer exposure to mycotoxins, the levels of deoxynivalenol (DON) and zearalenone (ZON) in cereals/cereal products and fumonisins B₁ and B₂ (FB₁ and FB₂) in maize are controlled by European Union legislation. Relatively few studies, however, have examined the loss of Fusarium mycotoxins during processing. The behaviour of FB₁, FB₂ and fumonisin B₃ (FB₃), DON and ZON during extrusion of naturally contaminated maize flour and maize grits is examined using pilot-scale equipment. DON and ZON are relatively stable during extrusion cooking but the fumonisins are lost to varying degrees. There is some loss of ZON when present in low concentrations and extruded at higher moisture contents. The presence of additives, such as reducing sugars and sodium chloride, can also affect mycotoxin levels. Moisture content of the cereal feed during extrusion is important and has a greater effect than temperature, particularly on the loss of fumonisins at the lower moistures. The effects are complex and not easy to explain, although more energy input to the extruder is required for drier materials. However, on the basis of these studies, the relationship between the concentration of Fusarium toxins in the raw and finished product is toxin- and process-dependent.     

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.     

Deoxynivalenol in wheat,maize, wheat flour and pasta: surveys in Hungary in 2008–2015

Among Fusarium mycotoxins, deoxynivalenol (DON) is the most common contaminant in case of cereals and cereal-based foods in Hungary. In this study, Hungarian wheat (n = 305), maize (n = 108), wheat flour (n = 179) and pasta (n = 226) samples were analysed (N = 818). The samples were collected during 2008–2015 in Hungary. Applied methods of analysis were enzyme-linked immunosorbent assay and liquid-chromatography coupled with a mass spectrometer. Results were compared and evaluated with Hungarian weather data. Among cereal samples, in 2011, wheat was contaminated with DON (overall average ± standard deviation was 2159 ± 2818 µg kg^?1), which was above the maximum limit (ML). In case of wheat flour and pasta, no average values above ML were found during 2008–2015, but higher DON contamination could be observed in 2011 as well (wheat flour: 537 ± 573 µg kg^?1; pasta: 511 ± 175 µg kg^?1).     

Arabinoxylan content and characterisation throughout the bread‐baking process

End‐use quality of wheat (Triticum aestivum L.) is influenced in a variety of ways by nonstarch polysaccharides, especially arabinoxylan (AX). The objective of this study was to track total and water‐extractable AX (TAX and WEAX, respectively) throughout the bread‐baking process, using wholemeal and refined flour. The TAX and WEAX content and the ratio of arabinose: xylose were assessed in flour, mixed dough, proofed dough and the bread loaf, which was separated into crumb, upper crust and bottom crust. Changes in TAX during the baking process differed between the refined flour and wholemeal samples, suggesting a change in the TAX availability which we ascribe to molecular interactions and heat treatment. WEAX content dramatically decreased during baking, suggesting that oxidative cross‐linkages rendered it unextractable. Higher levels of WEAX and lower levels of arabinose substitution were correlated with higher loaf volumes for refined flour among the hard wheat varieties. Having a better understanding of the importance of both WEAX content and arabinose substitution allows for directed breeding efforts towards improved hard wheat varieties for optimum bread baking.     

Fusarium mycotoxins in cereals harvested from Hungarian fields

The Fusarium mycotoxins deoxynivalenol (DON), zearalenone (ZEN) and T-2 frequently contaminate grain crops in Middle and Eastern Europe. In this survey, 116 cereal samples (maize, wheat, barley and oat) were examined for DON, ZEN and T-2 mycotoxins. Samples were collected from different areas in two Hungarian regions (North and South Transdanubia). The method of analysis was indirect competitive ELISA. Maize was the most contaminated grain regarding DON (86%), ZEN (41%) and T-2 (55%) toxins. The average results of the deoxynivalenol and zearalenone tests of maize proved to be significantly higher than those of barley or oat. DON was the most represented Fusarium mycotoxin followed by T-2 and ZEN. The examination of these mycotoxins would be necessary at a larger scale as to re-evaluate permissible levels, so increase of the monitoring programme would be advisable for the future.     

Occurrence of Fusarium T-2 and HT-2 toxins in oats from cultivar studies in Germany and degradation of the toxins during grain cleaning treatment and food processing

For the Fusarium trichothecene mycotoxins T-2 and HT-2, a combined (T-2 + HT-2) temporary tolerable daily intake (tTDI) of 0.06 µg kg^?1 body weight day^?1 was proposed at the European level in 2001 (Opinion of the Scientific Committee on Food). In the near future, maximum levels for these trichothecenes will be regulated by the European Commission as announced in EU (VO) 1881/2006. For the implementation of these maximum levels, more data on occurrence and behaviour of T-2 and HT-2 toxins in primary agricultural products as well as during cleaning treatment and food processing are needed. In the current work, we determined the T-2/HT-2 concentrations in four oat cultivars (Aragon, Dominik, Ivory, Pergamon) from ten different agricultural sites in Germany, grown in cultivar studies in 2007. The grains were de-hulled, oat meal was prepared, and bread with 20% oat meal and 80% wheat flour was baked. In the cereal-processing chain, samples were taken at various steps and subsequently analysed for their T-2/HT-2 content. We employed liquid chromatography-mass spectrometry (LC-MS) and an immunological screening method (enzyme-linked immunoabsorbant assay (ELISA)) for T-2/HT-2 determination. Detection limits were between 1 and 10 µg kg^?1 in different matrices. T-2/HT-2 concentrations determined by ELISA in oat samples from ten different agricultural sites in Germany were between 9 and 623 µg kg^?1. The median and 90th percentile were 48 and 191 µg kg^?1 T-2/HT-2, respectively. One site showed six times higher T-2/HT-2 levels than the other sites, where concentrations ranged from 322 to 623 µg kg^?1. In 80% of the samples the cultivars Pergamon and Ivory had the lowest concentration of T-2 and HT-2 toxins. Using LC-MS for T-2/HT-2 determination, cleaning of the raw material did not lead to significant reductions of T-2 and HT-2 levels, whereas de-hulling led to a reduction of over 90%. Boiling of oat meal produced from cleaned raw material to yield ‘porridge’ resulted in varying T-2/HT-2 levels in experimental replicates. No major reduction of T-2/HT-2 levels in cooked porridge was obtained. Standardized baking experiments using 20% oat meal showed that T-2 and HT-2 toxins are relatively stable during the baking process, probably due to their temperature stability.     

Fusarium head blight and associated mycotoxin occurrence on winter wheat in Luxembourg in 2007/2008

Fusarium head blight (FHB) is among the major causes of reduced quality in winter wheat and its products. In addition, the causal fungi produce a variety of toxins. A relatively high FHB infection rate in winter wheat was observed in 2007 and 2008 in Luxembourg. A fusariotoxin survey was carried out in 17 different geographical locations. Three groups of Fusarium mycotoxins (trichothecenes A and B and zearalenone) were analysed by a multi-detection HPLC–MS/MS method. Fusarium strains were also investigated by morphological and molecular methods. In addition, questionnaires relating to cultural practices were sent to the farmers managing the 17 fields investigated. FHB prevalence ranged from 0.3 to 65.8% (mean: 8.5%) in 2007 and from 0 to 24.5% (mean: 8.3%) in 2008. Results of morphological and molecular identification showed that the most common species isolated from diseased wheat spikes was F. graminearum (33.1%), followed by F. avenaceum (20.3%) and F. poae (17.8%). The chemical analysis revealed that 75% of the investigated fields were contaminated by deoxynivalenol (DON, range 0–8111 µg/kg). The preceding crop was highly and significantly correlated to the number of grains infected and had a significant impact on disease prevalence (p = 0.025 and 0.017, respectively, Fisher's F-test). A trend was found for maize as the preceding crop (p = 0.084, Tukey's test) to predict the amount of DON in the fields. This is the first report on the occurrence of DON and ZON in naturally infected wheat grains sampled from Luxembourg.     

Towards the development of peanut–wheat flour composite dough: Influence of reduced‐fat peanut flour on bread quality

The effect of partial substitution of wheat flour with reduced‐fat peanut flour at different levels (10, 20, 30, 40, and 50%) on physical parameters, proximate composition, sensory profile, and shelf stability of bread were investigated. Loaf volume, specific volume, and crumb density were significantly (p ≤ .05) reduced with increasing level of substitution with the peanut flour. Peanut flour had significant (p ≤ .05) improvement on the protein content and reduction in carbohydrate content of loaves. Consumers preferred the taste aroma and color of the peanut–wheat flour composite loaves at ≥20% peanut flour inclusion. Freshly baked composite peanut–wheat bread loaves with 10% level of peanut substitution had higher overall acceptability than 100% wheat flour formulation but less microbial stability during storage. Reduced fat‐peanut has potential application for improving the nutritional quality and shelf stability of wheat flour bread.

Practical applications

The demand for convenient alternative to conventional foods is on the increase with the dynamics of the world's social values, lifestyles, and demographic trends. Having peanut incorporated into dough (as one food system) will offer convenience to consumer and therefore add value to bread variety on market shelves. Assessing the influence of the peanut flour on bread quality provides first‐hand information that can facilitate optimization of the baking process toward commercial production of peanut–wheat flour bread.     

Thermal degradation of deoxynivalenol during maize bread baking

The thermal degradation of deoxynivalenol (DON) was determined at isothermal baking conditions within the temperature range of 100-250°C, using a crust-like model, which was prepared with naturally contaminated maize flour. No degradation was observed at 100°C. For the temperatures of 150, 200 and 250°C, thermal degradation rate constants (k) were calculated and temperature dependence of DON degradation was observed by using Arrhenius equation. The degradation of DON obeyed Arrhenius law with a regression coefficient of 0.95. A classical bread baking operation was also performed at 250°C for 70?min and the rate of DON degradation in the bread was estimated by using the kinetic data derived from the model study. The crust and crumb temperatures recorded during bread baking were used to calculate the thermal degradation rate constants (k) and partial DON degradations at certain time intervals. Using these data, total degradation at the end of the entire baking process was predicted for both crust and crumb. This DON degradation was consistent with the experimental degradation data, confirming the accuracy of kinetic constants determined by means of the crust-like model.     

BAKING PERFORMANCE AND CONSUMER ACCEPTABILITY OF RAW AND EXTRUDED COWPEA FLOUR BREADS

Cowpea flour was used to partially replace wheat flour in yeast bread, using automatic household‐type bread machines for mixing, proofing and baking. Loaves containing 15 or 30% extruded cowpea flour weighed more (683.4 g) than loaves from other treatments (641.1–652.6 g). The 100% wheat had the highest loaf volume (2.58 L) and the 30% extruded cowpea the lowest (1.64 L). Cowpea flour breads contained more protein (13.9–15.4%) than the 100% wheat (4.1% fat, 12.5% protein). Bread made with 15% extruded cowpea flour was not different (P < 0.05) from the all‐wheat control in sensory quality and acceptability. Hedonic ratings for the control and 15% extruded cowpea flour ranged from 6.6 (like slightly) to 7.4 (like moderately) for all sensory attributes. The least liked samples contained either 30% raw or 30% extruded cowpea flour, receiving ratings for all attributes ranging from 4.8 (disliked slightly) to 6.2 (liked slightly). Overall, 15% extruded cowpea flour demonstrated successful bread making performance without compromising sensory quality.     

Benefits of β-xylanase for wheat biomass conversion to bioethanol

BACKGROUND: The efficiency of bioethanol production from wheat biomass is related to the quality of end products as well as to safety criteria of co‐products such as distiller's dried grains with solubles (DDGS). The inclusion of a new biocatalyst for non‐starch polysaccharide degradation in fermentation processes could be one of the solutions. The objective of this study was to evaluate the influence of β‐xylanases in combination with traditional amylolytic enzymes on the efficiency of bioethanol production and DON detoxification during fermentation of Fusarium‐contaminated wheat biomass with high concentration of deoxynivalenol (DON; 3.95 mg kg^?1). RESULTS: The results showed that the negative effect of Fusarium spp. on yield and quality of bioethanol could be eliminated by the application of Trichoderma reesei xylanase in combination with amylolytic enzymes. This technological solution allowed to increase the concentration of ethanol in the fermented wort by 35.3% and to improve the quality of bioethanol by decreasing the concentrations of methanol, methyl acetate, isoamyl and isobutyl alcohols. Mass balance calculations showed that DDGS was the main source of DON contamination, comprising 74% of toxin found in wheat biomass. By using new enzyme combination for wheat biomass saccharification, a higher level of detoxification (41%) of DON was achieved during the fermentation process. CONCLUSION: The addition of Trichoderma reesei xylanase played a positive role in bioethanol production from Fusarium‐contaminated wheat biomass, indicating that the yeast‐growing medium was enriched during the enzymatic treatment. Copyright © 2011 Society of Chemical Industry