Antiviral Activity of Trichothecene Mycotoxins (Deoxynivalenol,Fusarenon-X,and Nivalenol) against Herpes Simplex Virus Types 1 and 2

The effect of trichothecene mycotoxins, deoxynivalenol (DON), fusarenon-X (FX) and nivalenol (NIV), on plaque formation of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) in HEp-2 cells was examined. The 50% effective concentrations (EC₅₀) of DON, FX, and NIV for HSV-1 plaque formation were 160, 56, and 120 ng/ml, respectively. Those for HSV-2 plaque formation were 94, 26, and 50 ng/ml, respectively. These three mycotoxins showed about 2-fold higher selectivity to HSV-2 than to HSV-1. Plaque formation of HSV-1 was not inhibited with trichothecenes at concentrations completely inhibiting plaque formation when cells were treated during virus adsorption period or 15 hr before infection. These results indicate that trichothecenes affect replication of HSV-1 after virus adsorption, but not before or during virus adsorption to the host cells.     

Cytotoxic and immunotoxic effects of Fusarium mycotoxins using a rapid colorimetric bioassay

A colorimetric MTT (tetrazolium salt) cleavage test was used to evaluate cytotoxicity of twenty-three Fusarium mycotoxins on two cultured human cell lines (K-562 and MIN-GL1) as well as their inhibitory effect on proliferation of phytohemagglutinin-stimulated human peripheral blood lymphocytes. The values of 50% inhibition of lymphocyte blastogenesis were very close to the 50% cytotoxic doses observed with the more sensitive cell line (MIN-GL1). T-2 toxin was the most cytotoxic with CD₅₀ and ID₅₀ values less than 1 ng/ml. Type A trichothecenes were the most cytotoxic followed by the type B trichothecenes; the non-trichothecenes were the least cytotoxic. The MTT cleavage test, in conjunction with cell culture, is a simple and rapid bioassay to evaluate cytotoxicity and immunotoxicity of Fusarium mycotoxins.Abbreviations Ac acetyl - ACU acuminatin - DAS diacetoxyscirpenol - DON deoxynivalenol - FUS fusarenon-X - HT-2 HT-2 toxin - MC mononuclear cell - MTT 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide - NEO neosolaniol - NIV nivalenol - NT-1 4,8-diacetoxy T-2 tetraol - PBS phosphate buffered saline - TAT-2T tetraacetoxy T-2 tetraol - T-2 T-2 toxin     

Natural Occurrence of 16 Fusarium Toxins in Grains and Feedstuffs of Plant Origin from Germany

A total of 220 samples comprising cereals, cereal byproducts, corn plants and corn silage as well as non-grain based feedstuffs was randomly collected during 2000 and 2001 from sources located in Germany and analysed for 16 Fusarium toxins. The trichothecenes scirpentriol (SCIRP), 15-monoacetoxyscirpenol (MAS), diacetoxyscirpenol (DAS), T-2 tetraol, T-2 triol, HT-2 and T-2 toxin (HT-2, T-2), neosolaniol (NEO), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivealenol (15-ADON), nivalenol (NIV) and fusarenon-X (FUS-X) were determined by gas chromatography/mass spectrometry. Zearalenone (ZEA) and α- and β-zearalenol (α- and β-ZOL) were analysed by high performance liquid chromatography with fluorescence and UV-detection. Detection limits ranged between 1 and 19 μg/kg. Out of 125 samples of a group consisting of wheat, oats, corn, corn byproducts, corn plants and corn silage only two wheat samples did not contain any of the toxins analysed. Based on 125 samples the incidences were at 2–11% for DAS, NEO, T-2 Triol, FUS-X, α- and β-ZOL, at 20–22% for SCIRP, MAS, T-2 tetraol and 3-ADON, at 44–74% for HT-2, T-2, 15-ADON, NIV and ZEA, and at 94% for DON. Mean levels of positive samples were between 6 and 758 μg/kg. Out of 95 samples of a group consisting of hay, lupines, peas, soya meal, rapeseed meal and other oilseed meals, 64 samples were toxin negative. DAS, T-2 triol, NEO and FUS-X were not detected in any sample. The incidences of DON and ZEA were at 14 and 23% respectively, those of the other toxins between 1–4%, mean levels of positive samples were between 5 and 95 μg/kg.     

Trichothecenes and Mycoflora in Wheat Harvested in Nine Locations in Buenos Aires Province, Argentina

A total of 120 freshly harvested wheat samples from the 2004 season in nine locations from Northern Buenos Aires Province, Argentina, were analysed for trichothecene natural occurrence and associated mycoflora, and for determining the influence of commonly used fungicide field treatment and the cultivar type on trichothecene contamination. The trichothecenes T-2 tetraol, T-2 triol, HT-2 and T-2 toxin (HT-2, T-2), diacetoxyscirpenol (DAS), nivalenol (NIV), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON) were analysed by gas chromatography and electron capture detection. Detection limits ranged from 4 to 20 μg/kg. The isolation frequencies of species were calculated. Alternaria alternata, Fusarium graminearum, Fusarium poae and Fusarium semitectum were the predominant fungal species identified as endogenous mycoflora. The type of cultivar and the fungicide field treatment did not affect significantly the trichothecene contamination. The trichothecenes type A detected were HT-2 and T-2 triol toxins and the type B were DON, NIV and 3-ADON. Based on 120 samples the incidences were 21.7% for 3-ADON, 22.5% for HT-2, 27.5% for T-2 triol and 85% for DON. NIV was confirmed in one sample. Mean levels of trichothecene positive samples were between 7 and 2788 μg/kg.     

Mycotoxins in horse feed

A total of 62 samples of commercial horse feed preparations (complementary feeds) containing cereal mixtures (“muesli” or mash, n = 39; pelleted feeds, n = 12), and plain horse feed grains (maize, n = 5; oats, n = 4; barley, n = 2) were purchased from 21 different producers/distributors from the German market. All samples were analysed by competitive enzyme immunoassays (EIA) for six different mycotoxins (mycotoxin groups). Analytes (detection limit, mean recovery) were: deoxynivalenol (DON, 10 μg/kg, 84%), zearalenone (ZEA, 5 μg/kg, 93%), fumonisin B₁ (FB₁, 2 μg/kg, 113%), T-2 toxin (T-2, 0.1 μg/kg, 71%), sum of T-2 + HT-2 toxin (T-2/HT2, 0.2 μg/kg, 97%), ochratoxin A (OTA, 0.2 μg/kg, 67%), and total ergot alkaloids (Generic Ergot Alkaloids “GEA”, 30 μg/kg, 132%). All samples contained DON (16–4,900 μg/kg, median 220 μg/kg), T-2/HT-2 (0.8–230 μg/kg, median 24 μg/kg), and T-2 (0.3–91 μg/kg, median 7 μg/kg). ZEA was detected in 98% of the samples (7–310 μg/kg, median 61 μg/kg). Most samples (94%) were positive for FB₁ (2–2,200 μg/kg, median 27 μg/kg). Ergot alkaloids were detected in 61% of samples (28–1,200 μg/kg, median 97 μg/kg), OTA was found in 42% of samples (0.2–4 μg/kg, median 0.35 μg/kg). The results demonstrate that a co-contamination with several mycotoxins is very common in commercial horse feed from the German market. The toxin concentrations were in most cases well below the levels which are usually considered as critical or even toxic. The highest mycotoxin concentrations were mostly found in single-grain cereal feed: the maximum values for DON and FB₁ were found in maize, the highest T-2/HT-2 toxin concentrations were found in oats, and the highest concentration of ergot alkaloids was found in barley. In composed feeds, no correlation between cereal composition and mycotoxin levels could be found.     

Occurrence of trichothecenes, zearalenone and ochratoxin A in cereals and mixed feed from central Lithuania

A total of 92 samples — 23 winter wheat, 12 summer barley, 5 oats and 52 mixed feed — were collected from a state factory in Kaunas, Lithuania and were analysed for the presence of trichothecenes, zearalenone (ZEN) and ochratoxin A (OA) using gas chromatography with electron capture detection and immunoaffinity column/high performance liquid chromatography with fluorescence and UV detections. Deoxynivalenol (DON), nivalenol (NIV), T-2 toxin and HT-2 toxin were detected at concentrations above 10 μg/kg in 68%, 48%, 38% and 8% of cereal samples, respectively, and in 98%, 88%, 12% and 8% of samples of mixed feed for swine and poultry. More than 10 μg/kg of zearalenone and ochratoxin A were found in 58% and 92% of the mixed feed samples, respectively. The highest concentrations of all analysed trichothecenes in Lithuanian mixed feed and cereal grains, with an exception of T-2 toxin in one oat lot and one sample of mixed feed and OA in two mixed feed samples, were lower than those reported as Lithuanian advisory or tolerance limits.     

Study on biodegradation of some A- and B-trichothecenes and ochratoxin A by use of probiotic microorganisms

In vitro biodegradation experiments were done using some probiotic microorganisms. DifferentSaccharomyces cerevisiae, Lactobacilli andBacilli strains were tested for their ability to degrade Nivalenol (NIV), Deoxynivalenol (DON), Diacetoxyscirpenol (DAS), T2-Toxin and Ochratoxin A (OTA). The concentrations of selected mycotoxins were in the range of natural occurring toxin contaminations (1ppm for NIV and DON, 500ppb for DAS and T-2 and 50ppb for OTA). No alteration of concentrations could be registered for the tested trichothecenes. The best results could be achieved in experiments with OTA by up to 94% detoxification. Influence of toxins on colony forming unit of all tested microorganisms were recorded. Especially T-2 toxin and DAS have a slowing effect on growth of some strains.     

Effect of 4-week feeding of deoxynivalenol- or T-2-toxin-contaminated diet on lipid peroxidation and glutathione redox system in the hepatopancreas of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.)

The purpose of study was to investigate the effects of T-2 toxin (4.11 mg T-2 toxin and 0.45 mg HT-2 toxin kg^?1 feed) and deoxynivalenol (5.96 and 0.33 mg 15-acetyl deoxynivalenol (DON)?kg^?1 feed) in 1-year-old common carp juveniles in a 4-week feeding trial. The exposure of mycotoxins resulted in increased mortality in both groups consuming mycotoxin-contaminated diet. Parameters of lipid peroxidation were not affected during the trial, and antioxidant defence also did not show response to oxidative stress; however, glutatione peroxidase activity slightly, but significantly, decreased in the T-2 toxin group. Glutathione S-transferase activity showed moderate decrease as effect of T-2 toxin, which suggests its effect on xenobiotic transformation. Reduced glutathione concentration showed moderate changes as effect of DON exposure, but T-2 toxin has no effect. Expression of phospholipid hydroperoxide glutathione peroxidase (GPx4) genes showed different response to mycotoxin exposure. T-2 toxin caused dual response in the expression of gpx4a (early and late downregulation and mid-term upregulation), but continuous upregulation was found as effect of deoxynivalenol. Expression of the other gene, gpx4b, was upregulated by both trichothecenes during the whole period. The results suggested that trichothecenes have some effect on free radical formation and antioxidant defence, but the changes depend on the duration of exposure and the dose applied, and in case of glutathione peroxidase, there was no correlation between expression of genes and enzyme activity.     

Intake estimates for trichothecene toxins of the population in Southwest Germany in 1998 and in 1999

The intake of theFusarium toxins deoxynivalenol (DON), nivalenol (NIV), HT-2 and T-2 toxin (HT-2, T-2), 3-, 15-acetyldeoxynivalenol (3-, 15- ADON), and fusarenon-X (FUS-X) was calculated for adults, children and babies, for an area of southwest Germany and two years (1998, 1999). Estimates were based on consumption data of bread and pasta by both adults and children and of infant food by babies, reported for the German population in a study on behalf of the European Union, and on toxin contents of a total of 208 samples of these commodities. No exceeding of the tolerable daily intake (TDI) of DON, NIV and the sum of HT-2 and T-2, as stated by the EU, was found for adults (70 kg body weight (BW)) and for babies (10 kg BW), independent of year and level of consumption. For children (20 kg BW) the intake of DON exceeded the TDI in 1998 for high, and in 1999 for both mean and high consumers. For both years the intake of the sum of HT-2 and T-2 was below the TDI following mean but above this value following high consumption. The intake of NIV was far below the TDI for both levels of ingestion. The daily intake of each of the three toxins 3-, 15- ADON and FUS-X was below 0.03, 0.11 and 0.05 μg/kg/BW for adults, children and babies, respectively. Presented at the 27^th Mykotoxin-Workshop, Dortmund. Germany, June 13–15, 2005     

Comparison of in vitro cytotoxicity of Fusarium mycotoxins,deoxynivalenol, T-2 toxin and zearalenone on selected human epithelial cell lines

Three human epithelial cell lines (CaCo-2, HEp-2 and HeLa) implicated as potential targets for three Fusarium toxins were tested for the extent of survival on exposure to increasing toxin concentration and incubation periods. Cytotoxicity assay using 3(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) was carried out with deoxynivalenol (DON), T-2 toxins and zearalenone (ZON) on CaCo-2, HEp-2 and HeLa cell lines. Of the three cell lines used, HeLa was the most sensitive, eliciting cell death after 2 days exposure at 100 ng ml^–1with T-2 toxin. HeLa was the only cell line to exhibit cytotoxicity towards ZON showing cell death at 1000 ng ml^–1after 2 days which increased to 4 days, showing substantial cell death at 200 ng ml^–1. HEp-2 was sensitive to DON showing cell death after 2 days (100 ng ml^–1) with complete cell death occurring at 200 ng ml^–1 after 4 days of exposure. Substantial cytoxicity of T-2 towards HEp-2 occurred after 2 days at 1000 ng ml^–1 and complete cell death occurred with 100 ng ml^–1 at day 4. The CaCo-2 cell line was generally resistant to the mycotoxins tested between 100 and 1000 ng ml^–1. This study shows that cytotoxicity of Fusarium toxins to epithelium cell lines is concentration- and time- dependant and results from ZON–HeLa interaction indicate possible cell type-mycotoxin specificity.     

Sources of variation in the analysis of trichothecenes in cereals by gas chromatography-mass spectrometry

The sources of variation In the analysis of trichothecenes in cereals by gas chromatography mass spectrometry (GC-MS) were studied and Identified. Ways to decrease some of the variations identified are presented and discussed. The method Is validated for deoxynivalenol, nivalenol, 3-acetyldeoxynivalenol, fusarenon-X, diacetoxyscirpenol, HT-2 toxin and T-2 toxin. The general variability in the performance of the GC-MS apparatus itself, as well as derivatisation, matrix effect, injection and quantification were identified to be the sources of variation. The use of internal standards was studied in order to decrease the variation due to problems with derivatisation and the overall sample preparation. Two derivatisation reagents were compared, one of which was found to be more effective for decreasing this variation. The calibrants prepared in the cereal extract reduced the variation due to the matrix effect and thus improved the quantification. “Constant Standards” were introduced in order to detect and decrease the variation caused by injection and the apparatus itself. The validation study proved that this analytical method for trichothecenes was adequately reliable and sensitive. The relative standard deviation varied between 3–9% and the recovery between 46–90% for the different trichothecenes determined. The limit of detection for a range of trichothecenes varied from 5 μg/kg to 15 μg/kg.     

Fusarium mycotoxins in forage maize — Detection and evaluation

The deoxynivalenol concentrations found in forage maize ranged between 0.24 and 14.29 mg/kg DM (detected by ELISA). When highly contaminated samples were analysed for deoxynivalenol by HPLC or LC-MS the resulting concentrations were in the mean about 50% lower. Furthermore, using LC-MS other type-A and type-B trichothecenes, zearalenone and α-zearalenol were found in these samples. The differences between ELISA and HPLC/LC-MS data for deoxynivalenol are assumed to result from cross-reactions of other trichothecenes with the antibodies used in ELISA and toxin losses from sample purification procedures needed for HPLC and LC-MS analysis. Presented at the 26^th Mykotoxin-Workshop in Herrsching, Germany, May 17–19, 2004     

Mycotoxins and fungi in wheat harvested during 1990 in test plots in the state of São Paulo,Brazil

Wheat from two cultivars with contrasting characteristics were harvested in ten experimental plots located in wheat producing areas of the State of São Paulo, Brazil. The samples (10 of each cultivar) were analyzed by a gaschromatographic method for deoxynivalenol (DON), nivalenol (NIV), diacetoxyscirpenol (DAS), toxins T-2 (T-2) and HT-2, T-2 tetraol, T-2 triol, and by a thin-layer chromatographic method for zearalenone (ZEN), aflatoxins B₁, B₂, G₁, G₂, ochratoxin A and sterigmatocystin. No mycotoxins were detected in 13 samples. DON was found in four samples (0.47–0.59 µg/g), NIV in three samples (0.16–0.40 µg/g), T-2 in two samples (0.40, 0.80 µg/g), DAS in one sample (0.60 µg/g), and ZEN in three samples (0.04–0.21 µg/g). The wheat samples were also examined for the incidence of fungi.Alternaria, Drechslera, Epicoccum andCladosporium were the prevailing genera. Among theFusarium spp.,F. semitectum was present in 19 samples andF. moniliforme in 18 samples. NoF. graminearum was isolated in the samples.Abbreviations DAS diacetoxyscirpenol - DON deoxynivalenol - NIV nivalenol - T-2 T-2 toxin - ZEN zearalenone     

Mycotoxins (trichothecenes, zearalenone and fumonisins) in cereals associated with human red-mold intoxications stored since 1989 and 1991 in China

Two corn powder samples implicated in the human food poisoning that occurred in Guangxi province in 1989, and eight wheat and two barley samples linked to an episode that involved about 130,000 people in gastrointestinal disorders in Anhui province in 1991 were analyzed for trichothecenes including deoxynivalenol (DON), nivalenol (NIV) and their esters, zearalenone (ZEA) and fumonisins (FMs) by gas chromatography/mass spectroscopy and high performance liquid chromatography, and T-2 toxin by enzyme-linked immunosorbent assays. DON was detected in all samples as a major trichothecene (16-51,450 microg kg(-1)), and NIV was in one corn, one barley and all wheat at relatively low levels (10-6935 microg kg(-1)). ZEA was found in all corn and barley, and six wheat samples (46-3079 microg kg(-1)). In addition, 3-acetyl-DON (2544 microg kg(-1)) and 15-acetyl-DON (2537 microg kg(-1)) were detected separately in one corn and one wheat sample. The highest levels of these mycotoxins were found in one wheat sample associated with the human intoxication in Anhui province. FMs in corn were below 1000 microg kg(-1). Risks of DON and ZEA on the people who consumed the causative cereals were assessed.     

Occurrence of type A,B and D trichothecenes in barley and barley products from the Bavarian market

Fifty-nine samples of barley and barley products were analysed for 18 trichothecene mycotoxins by a sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method (detection limits 0.062-0.70 μg/kg) after sample extract clean-up on MycoSep®-226 columns. The samples were collected in 2009 from barley processing facilities (mills and malt houses) and at wholesale and retail stage from the Bavarian market. The predominant toxins were T-2 toxin (T-2), HT-2 toxin (HT-2) and deoxynivalenol (DON). For all samples, the mean levels of T-2 and HT-2 were 3.0 μg/kg and 6.8 μg/kg with rates of contamination of 63% and 71%, respectively. The maximum values were 40 μg/kg for T-2 and 47 μg/kg for HT-2. The rate of contamination with DON was high (95%) with a low mean level of 23 μg/kg. The DON levels ranged between 3.4 to 420 μg/kg. For T-2 tetraol, a mean level of 9.2 μg/kg and a maximum level of 51 μg/kg with a rate of contamination of 71% were determined. NIV was detected in 69% of the samples with a mean level of 11 μg/kg and a maximum level of 72 μg/kg. Other type A and B trichothecenes were detected only in traces. Type D trichothecenes, fusarenon-X, verrucarol and 4,15-diacetylverrucarol were not detected in any sample. Winter barley and malting barley were the most contaminated groups of all samples in this study. In malting barley, the highest levels of contamination with type A trichothecenes were found. In contrast, winter barley showed the highest contamination with type B trichothecenes. The lowest mycotoxin concentrations were found in de-hulled and naked barley and in pearl barley.     

Effects of deoxynivalenol (DON) and related compounds on bovine peripheral blood mononuclear cells (PBMC) in vitro and in vivo

The Fusarium toxin deoxynivalenol (DON) often co-occurs along with the acetylated derivatives 3-acetyl-DON and 15-acetyl-DON in diets for ruminants. De-epoxy-DON is formed by rumen micro-organisms, while the acetylated DON derivatives might also undergo ruminal metabolism with de-epoxy-DON as an end product. However, despite the fact that de-epoxy-DON is the predominant substance finally absorbed, a complete degradation of the mother compounds can not be assumed for all feeding and metabolic situations of the cow, and thus raising the question of their possible post-absorptive effects. Hence, the aim of the study was to examine the effects of all four compounds on the concanavalin A stimulated proliferation of bovine peripheral blood mononuclear cells (PBMC) using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) as indicator in vitro and ex vivo. Among the DON-related compounds, DON and 15-acetyl-DON resulted in a similar IC₅₀ (i.e. the concentration where the proliferation was inhibited by 50%) of 0.5 μM, whereas 3-acetyl-DON was less toxic (IC₅₀ = 2.6 μM), while actually no IC₅₀ could be estimated for de-epoxy-DON which was characterized by a maximum inhibition of approximately 24% at the highest tested in vitro concentration of 18.29 μM. For the in vivo experiment, 14 Holstein cows were used and fed either an uncontaminated control diet (CON) or a diet contaminated with Fusarium toxins, with DON being the predominating toxin for 18 weeks when blood was collected for PBMC isolation and subsequent proliferation/viability assay. The complete diets for the CON and FUS group contained 0.4 and 4.6 mg DON/kg DM, respectively, at that time. Exposure of dairy cows to the FUS diet resulted in maximum serum de-epoxy-DON levels of 52 ng/ml (0.19 μM), while levels of the unmetabolized DON reached maximum levels of 9 ng/ml (0.03 μM). The PBMC of these cows were slightly less viable, by approximately 18% (p = 0.057), while stimulation capability was not decreased at the same time. Although de-epoxy-DON was characterized by the lowest in vitro toxicity among the tested DON-related compounds, there appeared to be a lower viability of the PBMC isolated from cows fed the FUS diet, which had nearly exclusively de-epoxy DON in serum beside slight traces of unmetabolized DON. Thus, the factors responsible for these apparent discrepancies need to be clarified.     

Natural multi-occurrence of mycotoxins in rice from Niger State,Nigeria

Twenty-one rice samples from field (ten), store (six) and market (five) from the traditional rice-growing areas of Niger State, Nigeria were analysed for aflatoxins (AFs), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), fumonisin B₁ (FB₁) and B₂ (FB₂), and patulin (PAT) by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) respectively. T-2 toxin was determined using TLC only. AFs were detected in all samples, at total AF concentrations of 28–372 μg/kg. OTA was found in 66.7% of the samples, also at high concentrations (134–341 μg/kg) that have to be considered as critical levels in aspects of nephrotoxicity. ZEA (53.4%), DON (23.8), FB₁ (14.3%) and FB₂ (4.8%) were also found in rice, although at relatively low levels. T-2 toxin was qualitatively detected by TLC in only one sample. Co-contamination with AFs, OTA, and ZEA was very common, and up to five mycotoxins were detected in a single sample. The high AF and OTA levels as found in rice in this study are regarded as unsafe, and multi-occurrences of mycotoxins in the rice samples with possible additive or synergistic toxic effects in consumers raise concern with respect to public health.     

The effect of vitamin E supplementation on reduction of lymphocyte DNA damage induced by T-2 toxin and deoxynivalenol in weaned pigs

The objective of the present study was to establish the effect of deoxynivalenol (DON) and T-2 toxin on lipid peroxidation, lymphocyte DNA fragmentation and immunoglobulin production in weaned pigs, and furthermore, to evaluate the potential of vitamin E (α-tocopheryl acetate) in prevention of toxin mediated changes. Forty-eight weaned castrated male crossbred pigs (mean live weight at the beginning of the experimental period was 11.7 kg) were randomly assigned to five experimental groups: control (without toxin and vitamin E), T-2 (3 mg/kg T-2 toxin), T-2 + E (3 mg/kg T-2 toxin + 100 mg/kg vitamin E), DON (4 mg/kg DON) and DON + E (4 mg/kg DON + 100 mg/kg vitamin E). After 14 days of treatment blood was collected for analysis. Lipid peroxidation was studied by assays of malondialdehyde (MDA), total antioxidant status (TAS) of plasma and erythrocyte glutathione peroxidase (GPx). DNA damage in lymphocytes was measured by comet assay. Serum immunoglobulin levels were evaluated by enzyme-linked immunosorbent assay (ELISA) and the hepatotoxicity was studied by measuring plasma liver enzyme levels (alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ-glutamyl-transferase (GGT). Production parameters of both DON groups were significantly impaired in comparison to the control. DON significantly increased the amount of DNA damage in lymphocytes by 28%. Moreover, the levels of TAS were lowered by addition of DON. T-2 toxin significantly impaired daily live weight gain and feed conversion, increased the amount of DNA damage in lymphocytes by 27%, decreased total serum IgG and did not alter plasma TAS. Plasma and 24-h urinary malondialdehyde (MDA) excretion rate and erythrocyte Gpx levels did not differ among the groups. Supplementation with vitamin E did not improve production parameters impaired by DON and T-2 toxin and only partially protected lymphocyte DNA from toxin impact. To our knowledge, these are the first data on genotoxic effects of moderate doses of DON and T-2 toxin on pig lymphocytes. The effect of DON and T-2 toxin on the immune system was reflected as a change in immunoglobulin synthesis, which might be toxin and species specific. According to other results no major induction of oxidative stress could be proven. Enhancement of antioxidant status with vitamin E in the case of DON and T-2 toxin intoxication can be beneficial for remaining the lymphocyte DNA integrity.     

Occurrence of Trichothecenes in feed and cereals in Finland

In 1985 82 samples of feed and food grain were analyzed for trichothecenes deoxynivalenol (DON), nivalenol (NV), diacetoxyscirpenol (DAS), T-2 toxin, HT-2 toxin and fusarenon-X (F-X). Trichothecenes were found in 77 of these samples. The highest amounts were DON 6300 ug/kg and DAS 1680 ug/kg.In 1986, in a corresponding study of 113 samples, trichothecenes were found in 110 samples. A new trichothecene in Finland, 3-acetyldeoxynivalenol (3-AcDON), was identified in 35 of these samples in concentration of 2–211 ug/kg.Analyzing methods were gas chromatography and GC-mass spectrometry. It is characteristic of the feed samples suspected of causing outbreaks in animals in Finland that several trichothecenes are often found in the same sample. As an example of this is a poultry feed with following results: DON 33 ug/kg, 3-AcDON 21 ug/kg, DAS 45 ug/kg, T-2 toxin 40 ug/kg, HT-2 toxin 12 ug/kg.     

Fusarium toxins in cereals grown in Switzerland

770 cereal samples of Swiss origin which were collected in various feed mills and cereal collection centres in the years 2000 – 2002 were assayed for Deoxynivalenol (DON) and zearalenone (ZEA). 137 samples were also assayed for T-2 toxin. The prevalence of DON and ZEA contamination was higher in cereals harvested in the rainy summer 2002 than in the previous years. T-2 toxin levels exceeding 100 μg/kg were found only in three oats samples. High levels ofFusarium toxins do not frequently occur in Swiss cereals. Presented at the 25^th Mykotoxin Workshop in Giessen, Germany, May 19–21, 2003