Concentrations Of Trichothecenes Research Articles

Experimental study on small molecule combinations inducing reprogramming of rat fibroblasts into functional neurons.

To establish a methodological system for reprogramming rat embryonic fibroblasts (REF) into chemically induced neurons (ciNCs) via small molecule compounds to provide safe and effective donor cells for treatment of neurodegenerative diseases. Based on the method established by PEI Gang's research group to directly reprogram human fibroblasts into neurons, the induction medium and maturation medium was optimized by replacing the coating solution, mitigating oxidative stress injury, adding neurogenic protective factors, adjusting the concentration of trichothecenes, performing small-molecule removal experiments, and carrying out immunofluorescence and Western blotting on cells at different stages of induction to validate the effect of induction. When the original protocol was used for induction, the cell survival rate was (34.24±2.77)%. After replacing the coating solution gelatin with matrigel, the cell survival rate increased to (45.41±4.27)%; after adding melatonin, the cell survival rate increased to (67.95±5.61)% and (23.43±1.42)% were transformed into neural-like cells; after adding the small molecule P7C3-A20, the cell survival rate was further increased to (76.27±1.41)%, and (39.72±4.75)% of the cells were transformed into neural-like cells. When the concentration of trichothecene was increased to 30 μmol/L, the proportion of neural-like cells reached (55.79±1.90)%; after the removal of SP600125, (86.96±2.15)% of the cells survived, and the rate of neural-like cell production increased to (63.43±1.60)%. With the optimized protocol, REF could be successfully induced into ciNC through the neural precursor cell stage, in which the neural precursor cells were able to highly express the neural precursor cell markers SRY-related HMG-box gene 2 (Sox2) and paired box 6 (Pax6) as well as neuron-specific marker tubulin 1 (Tuj1), while the expression of fiber-associated protein vimentin was reduced. After two weeks of induction of neural precursor cells in a maturation medium, most cells displayed neuronal-like cell morphology. The induced ciNCs were able to highly express the mature neuronal surface markers Tuj1 and microtubule-associated protein 2 (MAP2), while the expression of vimentin was reduced. The small molecule combinations optimized in this study can reprogram REF to ciNCs under normoxic conditions.

Impact of Environmental Conditions on the Concentrations of Trichothecenes, Their Glucosides, and Emerging Fusarium Toxins in Naturally Contaminated, Irradiated, and Fusarium langsethiae Inoculated Oats.

Trichothecenes produced by Fusarium species are commonly detected in oats. However, the ratios of the concentrations of free trichothecenes and their conjugates and how they are impacted by different interacting environmental conditions are not well documented. This study aims to examine the effect of water activity (0.95 and 0.98 aw) and temperature (20 and 25 °C) stress on the production of T-2 and HT-2 toxins, deoxynivalenol and their conjugates, as well as diacetoxyscirpenol (DAS). Multiple mycotoxins were detected using liquid chromatography-tandem mass spectrometry from 64 contaminated oat samples. The highest concentrations of HT-2-glucoside (HT-2-Glc) were observed at 0.98 aw and 20 °C, and were higher than other type A trichothecenes in the natural oats' treatments. However, no statistical differences were found between the mean concentrations of HT-2-Glc and HT-2 toxins in all storage conditions analysed. DAS concentrations were generally low and highest at 0.95 aw and 20 °C, while deoxynivalenol-3-glucoside levels were highest at 0.98 aw and 20 °C in the naturally contaminated oats. Emerging mycotoxins such as beauvericin, moniliformin, and enniatins mostly increased with a rise in water activity and temperature in the naturally contaminated oats treatment. This study reinforces the importance of storage aw and temperature conditions in the high risk of free and modified toxin contamination of small cereal grains.

The Effect of Ustilago maydis and Delayed Harvesting on A- and B-Type Trichothecene Concentrations in Maize Grain.

The aim of this study was to investigate whether, in the context of a higher incidence of Ustilago maydis and Fusarium spp. at optimal and delayed harvest times, a higher incidence of mycotoxin contamination in maize grains could be expected. The field experiment was carried out at the Lithuanian Research Centre for Agriculture and Forestry experimental fields over three consecutive years (2020-2022). Two maize hybrids (Duxxbury and Lapriora) with different FAO numbers were used. The experimental design in the field was a randomized complete block design. Harvesting took place at three different times: first at physiological maturity, and then 10 (±2) and 20 (±2) days after the first harvest. Each hybrid had four repetitions at different harvest times. The U. maydis infection was only detected in 2021 and after the first harvest cobs were further divided into four different groups with four repetitions: healthy cobs, cobs visually infected with Fusarium spp., cobs visually infected with common smut, and cobs visually infected with both pathogens. No U. maydis-damaged maize cobs were found in 2020 and 2022. The levels of Fusarium microscopic fungi in maize grains were also from 4 to 16 times higher in 2021 than in 2020 and 2022. Harvest delays in 2020 led to a significant deoxynivalenol concentration increase in the Duxxbury hybrid and an HT-2 concentration increase in the Lapriora hybrid. In 2021, deoxynivalenol, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, and HT-2 concentrations significantly rose in both hybrids, but the T-2 concentration significantly increased only in the Lapriora hybrid. Deoxynivalenol concentrations were, respectively, 110 and 14.6 times higher than in cobs only infected with Fusarium spp. or U. maydis. Concentrations of 15-acetyl-deoxynivalenol were, respectively, 60, 67, and 43 times higher than in asymptomatic cobs and cobs only infected with Fusarium spp. or U. maydis. Cobs contaminated with both pathogens also had higher concentrations of 3-acetyl-deoxynivalenol. T-2 and HT-2 were detected in maize grains harvested from cobs infected only with Fusarium spp. The presence of U. maydis and Fusarium fungi in maize cobs, along with harvest delays, led to significant increases in mycotoxin concentrations, highlighting the importance of timely harvesting and pathogen management to mitigate mycotoxin contamination in maize grains.

Effect of Crop Rotation on Fusarium Mycotoxins and Fusarium Species in Cereals in Sichuan Province (China).

The present study was performed to evaluate the effect of crop rotation on Fusarium mycotoxins and species in cereals in Sichuan Province. A total of 311 cereal samples were randomly collected and analyzed from 2018 to 2019 in Sichuan Province. The results of mycotoxin analysis showed that the major trichothecene mycotoxins in Sichuan Province were nivalenol (NIV) and deoxynivalenol (DON), and the mean concentration of total trichothecenes (including NIV, fusarenone X [4ANIV], DON, 3-acetyldeoxynivalenol [3ADON], and 15-acetyldeoxynivalenol [15ADON]) in wheat was significantly higher than that in maize and rice. The concentration of total trichothecenes in the succeeding crops was significantly higher than that in the previous crops. In addition, wheat grown after maize had reduced incidence and concentration of trichothecene mycotoxins compared with that grown after rice, and ratooning rice grown after rice had increased incidence and concentration of trichothecene mycotoxins. Our data indicated that Fusarium asiaticum with the NIV chemotype was predominant in wheat and rice samples, while the number of the NIV chemotypes of F. asiaticum and Fusarium meridionale and the 15ADON chemotype of Fusarium graminearum in maize were almost the same. Although the composition of Fusarium species was affected by crop rotations, there were no differences when comparing the same crop rotation except for the maize-wheat rotation. Moreover, the same species and chemotype of Fusarium strains originated from different crops in various rotations, but there were no significant differences in pathogenicity in wheat and rice. These results contribute to the knowledge of the effect of crop rotation on Fusarium mycotoxins and species affecting cereals in Sichuan Province, which may lead to improved strategies for control of Fusarium mycotoxins and fungal disease in China.

Trichothecenes in Food and Feed, Relevance to Human and Animal Health and Methods of Detection: A Systematic Review.

Trichothecene mycotoxins are sesquiterpenoid compounds primarily produced by fungi in taxonomical genera such as Fusarium, Myrothecium, Stachybotrys, Trichothecium, and others, under specific climatic conditions on a worldwide basis. Fusarium mold is a major plant pathogen and produces a number of trichothecene mycotoxins including deoxynivalenol (or vomitoxin), nivalenol, diacetoxyscirpenol, and T-2 toxin, HT-2 toxin. Monogastrics are sensitive to vomitoxin, while poultry and ruminants appear to be less sensitive to some trichothecenes through microbial metabolism of trichothecenes in the gastrointestinal tract. Trichothecene mycotoxins occur worldwide however both total concentrations and the particular mix of toxins present vary with environmental conditions. Proper agricultural practices such as avoiding late harvests, removing overwintered stubble from fields, and avoiding a corn/wheat rotation that favors Fusarium growth in residue can reduce trichothecene contamination of grains. Due to the vague nature of toxic effects attributed to low concentrations of trichothecenes, a solid link between low level exposure and a specific trichothecene is difficult to establish. Multiple factors, such as nutrition, management, and environmental conditions impact animal health and need to be evaluated with the knowledge of the mycotoxin and concentrations known to cause adverse health effects. Future research evaluating the impact of low-level exposure on livestock may clarify the potential impact on immunity. Trichothecenes are rapidly excreted from animals, and residues in edible tissues, milk, or eggs are likely negligible. In chronic exposures to trichothecenes, once the contaminated feed is removed and exposure stopped, animals generally have an excellent prognosis for recovery. This review shows the occurrence of trichothecenes in food and feed in 2011–2020 and their toxic effects and provides a summary of the discussions on the potential public health concerns specifically related to trichothecenes residues in foods associated with the exposure of farm animals to mycotoxin-contaminated feeds and impact to human health. Moreover, the article discusses the methods of their detection.

The influence of the conditions of acquisition and storage of table eggs on changes in their quality and the presence of mycobiota and Fusarium mycotoxins

The study showed that the hen housing system and egg storage conditions had significant influence on differences in the qualitative traits of eggs. The most rapid qualitative traits changes were observed in the eggs laid by the hens kept in the free-range and litter systems. After the first week of storage fungi were found on the eggshell surface but significant growth of fungi on the eggshell surface was noted after 3 wk, regardless of the storage conditions. Higher humidity during egg storage causes the dynamic development of fungi in the albumen. The principal component analysis proved the dependency between ergosterol and the trichothecene concentration in the albumen. Mycotoxins in the albumen indicate the presence of pathogenic species of microscopic fungi. The albumen of the eggs laid by the hens kept in cages had the lowest concentration of mycotoxins. The results of the study showed that the conditions of the henhouse environment significantly influenced the initial contamination of eggshells. Egg storage conditions, i.e., high air humidity, favor the development of fungi on the eggshell surface, affect the dynamics of fungal growth into the albumen and the production of mycotoxins.

Mycotoxin Production by Stachybotrys chartarum on Water-Damaged Building Materials

The major question after a flooding event is whether to remove or remediate the building materials so that potentially harmful mold growth and their by-products cannot cause serious health problems for susceptible individuals. The purpose of this study was to determine the growth of Stachybotrys chartarum and corresponding production of macrocyclic trichothecenes on different components of a residential wall up to 65 days after a simulated flood event. Small-scale residential walls constructed of fiberglass batt insulation, oriented strandboard, gypsum wallboard, and lumber were destructively sampled at four time points. All four building materials contained notable levels of macrocyclic trichothecenes on all collection days. The highest concentrations of macrocyclic trichothecenes were on the paper siding of the gypsum wallboard, followed by the paper siding of the batt insulation and wood lumber. There was a significant increase in trichothecene concentration over time, particularly on the gypsum. The DNA concentrations of the mold were significantly higher on the batt insulation than on the wood products, and the mold concentrations also increased over time on the batt insulation and gypsum, but not on the wood products. It was concluded that if a flooding event should occur, the insulation and gypsum should be removed from the home and the remaining materials should be remediated.

Microbial detoxification of eleven food and feed contaminating trichothecene mycotoxins

BackgroundContamination of agricultural commodities with multiple trichothecene mycotoxins, produced by toxigenic Fusarium species, is a food safety issue, which greatly affects grain production and marketing worldwide. Importantly, exposure to multiple trichothecenes may increase toxicity in animals due to their synergistic and/or additive effects. To address the problem this study aimed to achieve a novel biological trait capable of detoxifying various food and feed contaminating trichothecenes under aerobic and anaerobic conditions and wide range of temperatures.ResultsA highly enriched microbial consortium (called DX100) capable of transforming eleven trichothecenes to significantly less toxic de-epoxy forms was achieved after prolonged incubation of soil microbial culture with 200 μg/mL deoxynivalenol (DON). DX100 demonstrated de-epoxidation activity under aerobic and anaerobic conditions, a greater range of temperatures and around neutral pH. The consortium contains 70% known and 30% unknown bacterial species, dominated by Stenotrophomonas species. Probably novel bacteria including strains of Stenotrophomonas and Alkaliphilus-Blautia species complex could be involved in aerobic and anaerobic de-epoxidation of trichothecenes, respectively. DX100 showed rapid and stable activity by de-epoxidizing 100% of 50 μg/mL deoxynivalenol at 48 h of incubation and retaining de-epoxidation ability after 100 subcultures in mineral salts broth (MSB). It was able to de-epoxidize high concentration of DON (500 μg/mL), and transformed ten more food contaminating trichothecenes into de-epoxy forms and/or other known/unknown compounds. Microbial de-epoxidation rate increased with increasing trichothecene concentrations in the broth media, suggesting that DX100 maintains a robust trichothecene detoxifying mechanism. Furthermore, the nature of microbial de-epoxidation reaction and inhibition of the reaction by sodium azide and the finding that bacterial cell culture lysate retained activity suggests that certain cytoplasmic reductases may be responsible for the de-epoxidation activity.ConclusionsThis study reports the enrichment procedure for obtaining an effective and stable microbial consortium DX100 capable of de-epoxidizing several food contaminating trichothecene mycotoxins. DX100, which has de-epoxidation ability under wide range of conditions, represents a unique enzymatic source which has great industrial potential for reducing contamination of foods/feeds with multiple trichothecenes, and minimizing their synergistic/additive cytotoxic effects on consumer health.

Pathogenicity of Fusarium graminearum and F. meridionale on soybean pod blight and trichothecene accumulation

Soybean (Glycine max) is the most important crop in Argentina. At present Fusarium graminearum is recognized as a primary pathogen of soybean in several countries in the Americas, mainly causing seed and root rot and pre‐ and post‐emergence damping off. However, no information about infections at later growth stages of soybean development and pathogenicity of F. graminearum species complex is available. Therefore, the objectives of this study were to compare the pathogenicity of F. graminearum and F. meridionale isolates towards soybean under field conditions and to evaluate the degree of pathogenicity and trichothecene production of these two phylogenetic species that express different chemotypes. Six isolates of F. graminearum and F. meridionale were evaluated during 2012/13 and 2013/14 soybean growing seasons for pod blight severity, percentage of seed infected in pods and kernel weight reduction. The results showed a higher aggressiveness of both F. graminearum and F. meridionale species during the 2013/14 season. However, the differences in pathogenicity observed between the seasons were not reflected in a distinct trichothecene concentration in soybean seeds at maturity. Fusarium meridionale isolates showed similar pathogenicity to F. graminearum isolates but they were not able to produce this toxin in planta during the two field trials.

Effect of environmental factors on Fusarium population and associated trichothecenes in wheat grain grown in Jiangsu province, China

The present study was performed to identify prevailing Fusarium species and the environmental factors affecting their frequencies and the contamination of grain with major mycotoxins in Jiangsu province. The precipitation levels were 184.2mm, 156.4mm, and 245.8mm in the years 2013–2015, respectively, and the temperature fluctuated by an average of 10.6±7.2°C in 2013, 10.9±7.2°C in 2014, and 10.6±6.3°C in 2015. Co-occurrence of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3ADON), and 15-acetyldeoxynivalenol (15ADON) were observed in wheat. The average concentrations of DON were 879.3±1127.8, 627.8±640.5, and 1628.6±2,168.0μg/kg in 2013–2015, respectively. The average concentrations of 3ADON were 43.5±59.0, 71.2±102.5, and 33.5±111.9μg/kg in 2013–2015, respectively. We found that the average concentration of DON in wheat was positively correlated with precipitation (r=0.998, p<0.01), and 3ADON was negatively correlated with precipitation (r=−0.887, p<0.05). However, there was no correlation between precipitation and 15ADON or nivalenol (NIV). The differences in temperature were not as significant as the differences in rainfall amount over a short time period. Therefore, there were no correlations between temperature and the concentrations of trichothecenes, excluding 3ADON (r=0.996, p<0.01). Our data indicated that Fusarium asiaticum is the primary pathogenic fungus prevalent in the Fusarium head blight disease nursery. The trichothecene chemotype composition differed between Fusarium graminearum sensu stricto (s. str.) and F. asiaticum isolates. The 3ADON chemotype was found only among strains of F. asiaticum. The NIV chemotype was not observed among strains of F. graminearum, while the 15ADON chemotype represented 100% of the F. graminearum strains collected. The results of this study indicated no correlations between environmental conditions and the species or genetic chemotype composition of pathogens in Jiangsu province in 2013–2015.

Changes in contents of trichothecenes during commercial grain milling

Abstract The aim of this study was to determine changes in contamination with mycotoxins group of trichothecenes in naturally infested grain samples of 4 cereal species at every stage of the technological process. Samples were collected from the technological lines of a mill located in Poland in the years 2006–2008. They were analyzed in terms the concentration of trichothecenes A and B. The mean concentrations of total mycotoxins, rye grain contained the greatest amounts of these metabolites at 0.080 mg/kg, barley grain had a significantly lower total trichothecene concentration of 0.052 mg/kg. Mean contents of total toxins in oat and wheat grains were the lowest, did not differ significantly and amounted to 0.015 mg/kg and 0.014 mg/kg, respectively. This study showed a significant effect of the milling process on levels of mycotoxins in the final product.

Relationships between Genetic Diversity and Fusarium Toxin Profiles of Winter Wheat Cultivars.

Fusarium head blight is one of the most important and most common diseases of winter wheat. In order to better understanding this disease and to assess the correlations between different factors, 30 cultivars of this cereal were evaluated in a two-year period. Fusarium head blight resistance was evaluated and the concentration of trichothecene mycotoxins was analysed. Grain samples originated from plants inoculated with Fusarium culmorum and naturally infected with Fusarium species. The genetic distance between the tested cultivars was determined and data were analysed using multivariate data analysis methods. Genetic dissimilarity of wheat cultivars ranged between 0.06 and 0.78. They were grouped into three distinct groups after cluster analysis of genetic distance. Wheat cultivars differed in resistance to spike and kernel infection and in resistance to spread of Fusarium within a spike (type II). Only B trichothecenes (deoxynivalenol, 3-acetyldeoxynivalenol and nivalenol) produced by F. culmorum in grain samples from inoculated plots were present. In control samples trichothecenes of groups A (H-2 toxin, T-2 toxin, T-2 tetraol, T-2 triol, scirpentriol, diacetoxyscirpenol) and B were detected. On the basis of Fusarium head blight assessment and analysis of trichothecene concentration in the grain relationships between morphological characters, Fusarium head blight resistance and mycotoxins in grain of wheat cultivars were examined. The results were used to create of matrices of distance between cultivars – for trichothecene concentration in inoculated and naturally infected grain as well as for FHB resistance Correlations between genetic distance versus resistance/mycotoxin profiles were calculated using the Mantel test. A highly significant correlation between genetic distance and mycotoxin distance was found for the samples inoculated with Fusarium culmorum. Significant but weak relationships were found between genetic distance matrix and FHB resistance or trichothecene concentration in naturally infected grain matrices.

A comparison of contents of group A and B trichothecenes and microbial counts in different cereal species

Concentrations of trichothecenes and the amounts of microbial biomass were compared in grain of bread wheat, durum wheat, triticale, rye, oat and barley. Grain samples came from lines regionalised in Poland grown under identical climatic and agricultural conditions in 2007. Among the six analysed cereals, the highest mean concentration of toxic metabolites of 151.89 µg/kg was found for grain of Triticum durum, whereas the lowest was for barley grain (25.56 µg/kg). The highest contamination with microscopic fungi was recorded in case of barley and rice grain (mean concentrations of ergosterol were 12.53 mg/kg and 11.24 mg/kg, respectively). In case of the analysed cereals, the total microbial biomass expressed in the amount of ATP turned out to be the highest (4.7 × 105 relative light units [RLU]) for rye and oat (3.2 × 105 RLU). The results of the applied classical discrimination analysis indicate a significant diversification of species in terms of all the 11 analysed metabolites.

Bioherbicidal activity from washed spores of Myrothecium verrucaria

The fungal plant pathogen, Myrothecium verrucaria, is highly virulent to several important weed species and has potential utility as a bioherbicide. However the production of macrocyclic trichothecene mycotoxins by this fungus presents significant safety concerns. It was discovered that trichothecenes are removed from M. verrucaria spores by repeated washes with water. These washed spores retained bioherbicidal efficacy against kudzu when tested in field trials and on sicklepod when tested under greenhouse conditions. Changes in the growth medium combined with washing spores with water resulted in greater than 95% reduction in roridin A and verrucarin A. Washing spores reduced trichothecene concentrations in spore preparations with no significant effect on plant biomass reduction, thus demonstrating the possibility of M. verrucaria formulations with improved safety to researchers, producers and applicators.

Macrocyclic trichothecene production and sporulation by a biological control strain of Myrothecium verrucaria is regulated by cultural conditions

Myrothecium verrucaria is a pathogen of several invasive weed species, including kudzu, and is currently being evaluated for use as a bioherbicide. However, the fungus also produces macrocyclic trichothecene mycotoxins. The safety of this biological control agent during production and handling would be improved if an inoculum could be produced without concomitant accumulation of macrocyclic trichothecenes. Sporulation and trichothecene production by M. verrucaria was evaluated on standard potato dextrose agar (PDA) and a series of complex and defined media. Sporulation on PDA and on agar media with nitrogen as ammonium nitrate or potassium nitrate was more than ten-fold greater then sporulation on the medium with ammonium sulphate as the nitrogen source. Accumulation of macrocyclic trichothecenes was strongly affected by the media composition, with higher levels often associated with higher carbon content in the media. Overall, incubation in continuous darkness resulted in higher macrocyclic trichothecene concentrations. Results support the hypothesis that accumulation of macrocyclic trichothecenes by this fungus can be altered by manipulating carbon and nitrogen sources. Furthermore, the biosynthesis of these mycotoxins may be independent of sporulation, demonstrating that the bioherbicide can be readily produced on solid substrates while simultaneously yielding conidia that are less threatening to worker safety. A more detailed implementation of the concepts demonstrated in this study will facilitate the safe and economical production of this bioherbicide.

Concentrations of Some Metabolites Produced by Fungi of the Genus Fusarium and Selected Elements in Spring Spelt Grain

ABSTRACTThe aim of this study was to determine the content of selected elements and metabolites produced by fungi of the genus Fusarium in spelt (Triticum spelta L.) grain and husks and common wheat (T. aestivum L.) grain. Concentrations of trichothecenes, a volatile metabolite trichodiene (TRICH), as well as ergosterol (ERG) and adenosine 5′‐triphosphate (ATP) (a total microbial biomass indicator), were assessed. Toxin concentrations in spelt grain and husks harvested in 2003 and 2004 were comparable. Average deoxynivalenol concentrations reached 450 and 523 μg/kg in grain and 2,162 and 855 μg/kg in husks, respectively. Spelt grain, in comparison with common wheat grain, contained significantly higher concentrations of P, S, Mg, Zn, and Cu and a lower concentration of Al, whereas the concentrations of Ca, Fe, and Pb were significantly higher in the husks than in the grain of this cereal. A comparison of concentrations of Fusarium spp. metabolites in the grain of spelt and common wheat showed that the total concentration of mycotoxins and TRICH was slightly lower in T. spelta, whereas common wheat grain contained lower concentrations of ERG and ATP. The obtained results indicate that spelt husks contain considerable concentrations of trichothecenes.

Contents of trichothecenes in oats during official variety, organic cultivation and nitrogen fertilization trials in Finland

Natural toxins, such as mycotoxins, have emerged as a significant factor affecting the safety image of cereal grains as a raw material for the food and feed industry. The aim of the present study was to investigate the contents of trichothecenes in representative samples of oats during official variety, nitrogen fertilization and organic farming trials in Finland, 1997–1999. Further objectives were to promote industry and commerce by selection of high-quality oat varieties for various applications. The official variety trials conducted at 8–10 locations were managed following standard protocol. There were 2 types of agronomy trial, the first included comparison of oat cultivars grown in conventional and organic farming systems at 6 locations, and the second used 5 nitrogen rates (0, 40, 80, 120 and 160 kg N ha-1) at 2 locations. Regardless of wet cold summer occurring in Finland during 1998, the concentrations of Fusarium toxins were lowest during this 3 year monitoring period. More mycotoxins were produced during the warm, dry summers of 1997 and 1999 than in 1998. In all, 55% of the oat samples in the official variety trials contained deoxynivalenol (DON) within the range of 50– 896 µ g kg-1. The differences in DON concentrations between organic and conventional cultivation were small. The results showed also that the use of various nitrogen fertilization levels only slightly affected the trichothecene concentrations. The contents of trichothecenes in Finnish grains appeared to be similar to or lower than those reported earlier in the Northern Hemisphere.;

Mechanisms of deoxynivalenol-induced gene expression and apoptosis

Fusarium infection of agricultural staples such as wheat, barley and corn with concurrent production of deoxynivalenol (DON) and other trichothecene mycotoxins is an increasingly common problem worldwide. In addition to its emetic effects, chronic dietary exposure to DON causes impaired weight gain, anorexia, decreased nutritional efficiency and immune dysregulation in experimental animals. Trichothecenes are both immunostimulatory or immunosuppressive depending on dose, frequency and duration of exposure as well as type of immune function assay. Monocytes, macrophages, as well as T- and B-lymphocytes of the immune system can be cellular targets of DON and other trichothecenes. In vitro exposure to low trichothecene concentrations upregulates expression both transcriptionally and post-transcriptionally of cytokines, chemokines and inflammatory genes with concurrent immune stimulation, whereas exposure to high concentrations promotes leukocyte apoptosis with concomitant immune suppression. DON and other trichothecenes, via a mechanism known as the ‘ribotoxic stress response’, bind to ribosomes and rapidly activate mitogen-activated protein kinases (MAPKs). The latter are important transducers of downstream signalling events related to immune response and apoptosis. Using cloned macrophages, two critical upstream transducers of DON-induced MAPK activation have been identified. One transducer is double-stranded RNA (dsRNA)-activated protein kinase (PKR), a widely expressed serine/threonine protein kinase that can be activated by dsRNA, interferon and other agents. The other transducer is haematopoetic cell kinase (Hck), a non-receptor associated Src oncogene family kinase. Pharmacological inhibitors and gene suppression studies have revealed that Hck and PKR contribute to DON-induced gene expression and apoptosis. PKR, Hck and other kinases bind to the ribosome and are activated following DON interaction. Future studies will focus on the sequence of molecular events at the ribosome level that drive selective activation of these upstream kinases.

Natural occurrence of 16 Fusarium toxins in edible oil marketed in Germany

A total of 110 samples of edible oil marketed in Germany were analysed for 13 trichothecene toxins of the A- and B-type by gas chromatography/mass spectrometry, and for zearalenone (ZEA) and its derivatives by high performance liquid chromatography with fluorescence and UV-detection. Fusarium toxins were found in oil from soybean, sunflower and corn germ with up to 14 samples positive for at least one of the toxins 4, 15-diacetoxyscirpenol, HT-2 toxin, T-2 toxin, deoxynivalenol, 15-acetyldeoxynivalenol, ZEA and α-zearalenol. Scirpentriol, 15-monoacetoxyscirpenol, T-2 tetraol, T-2 triol, neosolaniol, 3-acetyldeoxynivalenol, nivalenol, fusarenon-X and β-zearalenol were not found. Trichothecene concentrations did not exceed 116 μg/kg, whereas up to 1730 μg/kg of ZEA were found in corn germ oil. Toxins were detected in refined and non-refined samples as well as in those of conventional and ecological production.

A rapid and sensitive cytotoxicity screening assay for trichothecenes in cereal samples

The application of a bioassay for toxicity screening of cereal samples has been studied. The sensitive Swiss mouse 3T3 fibroblasts (3T3 cells) were used to determine the cytotoxicity of Fusarium trichothecenes T-2 toxin (T-2), HT-2 toxin (HT-2), deoxynivalenol (DON) and nivalenol (NIV) in cereal samples. Clean-up of the sample extracts was performed using the MycoSep™ #225 column and the cytotoxic effects of the samples were determined using the 5-bromo-2′-deoxyuridine (BrdU) bioassay assessing DNA-synthesis. Trichothecene-free samples of wheat, wheat bran and barley had no toxic effect on the 3T3 cells at sample concentrations up to 400 mg/ml cell culture medium while a few oat samples had an inhibiting effect on the DNA-synthesis. IC 50 values (50% response compared with untreated cells) of trichothecene-free wheat and oats spiked with T-2, HT-2, DON and NIV were similar to IC 50 values of pure toxins. A cytotoxicity screening of 33 different cereal samples showed that all samples contaminated with a trichothecene concentration higher than the IC 50 values of T-2, HT-2, DON and NIV had a clear inhibiting effect on DNA-synthesis of the 3T3 cells. The culture of 3T3 cells on microtiter plates in combination with the simple and rapid sample clean-up using the MycoSep™ #225 column provides a sensitive and easy method for the screening of trichothecenes in cereal samples.