Fusaria and Fusarium toxins in New Zealand maize plants

A time course study was made of the development of Fusarium infection and the appearance of the three Fusarium toxins, nivalenol (NV), deoxynivalenol (DON) and zearalenone (ZEN), in various fractions of maize plants from two sites in New Zealand, one in the Manawatu region and one in the Waikato. Fusarium infection was seen in leaf axil fractions in January, at the time of tassel emergence, and was detectable in stalks, leaf blades, rachis and peduncles during February and in kernels in April. NV, DON and ZEN were only detectable some time after infection was demonstrable. NV, in high concentrations relative to DON (up to 287 mg/kg for NV and up to 8 mg/kg for DON), was found in fractions from the Manawatu site where F. crookwellense and F. culmorum were the predominant toxigenic species. NV and DON at similar levels (up to 25 mg/kg) were found in fractions from the Waikato site at which F. graminearum and F. subglutinans predominated. Highest levels of NV and DON were in rachis and peduncle. ZEN was found most consistently in leaf axils and blades at both sites (up to 8 mg/kg at the Manawatu site and up to 75 mg/kg at the Waikato site) but at times there were high levels in rachis fractions (up to 417 mg/kg at the Manawatu site). This revised version was published online in June 2006 with corrections to the Cover Date.     

Modified use of a commercial ELISA kit for deoxynivalenol determination in rice and corn silage

The analysis of deoxynivalenol (DON) in silage samples using enzyme-linked immunosorbent assay (ELISA) often leads to an overestimation. To better analyze DON in rice and corn silages using a commercially available ELISA kit, a cleanup method using a MultiSep #226 column was developed. As a result, overestimation of DON by the influence of specific cross-reaction with acetyldeoxynivalenol (AcDON) was confirmed. In samples where AcDON was not detected by liquid chromatography with mass spectrometry (LC-MS), no samples showed a significant difference (P?<?0.05) in DON amounts between ELISA with cleanup and LC-MS analysis. For the recovery study, blank silage was spiked with 0.5 or 1.0 mg/kg DON. The mean recoveries of DON determined by ELISA with cleanup and LC-MS analysis were 112 and 96 %, respectively, and the relative standard deviation for the repeatability (RSD_r) were 8.2 and 9.8 %, respectively. No samples showed a significant difference (P?<?0.05) in DON concentration determined by either ELISA or LC-MS analysis. A collaborative study to validate this rapid method was carried out using four samples, two rice and two corn silage, by 10 participating laboratories. Each sample was analyzed using blind duplicates. The mean values of DON detected were 1.5–2.3 mg/kg, RSD_r and the relative standard deviation for the reproducibility (RSD_R) were 4.1–12.7 and 7.6–23.4 %, respectively, and the HorRat values were 0.5–1.6. Therefore, the overestimation of DON by the influence of nonspecific cross-reaction with sample matrix was reduced by the cleanup method using a MultiSep #226 column, and analysis of DON in silage was improved. This use of this method for estimation of DON contamination in silage allows rapid detection at the place of use that is likely to result in improved animal health.     

Fusarium toxin residues in physiological samples of piglets

Physiological samples of 100 piglets fed diets containing 0.01, 0.06, 0.15, 0.22 and 0.42 mg ZON and 0.2, 0.8, 1.0, 1.9 and 3.9 mg DON per kg over a period of 35±1.5 days were investigated for concentrations of deoxynivalenol (DON) and zearalenone (ZON) and their metabolites. DON was detected in serum, bile and urine in increasing concentrations corresponding to the diet contamination. The metabolite de-epoxy-DON was detected only in urine. The DON contamination of the diet was closely reflected by the serum concentrations of the piglets. ZON and its metabolite α-zearalenol were detected in bile fluid, liver and urine, while β-zearalenol was only detected in bile fluid. In serum neither ZON nor its metabolites were found. The total concentration of ZON plus its metabolites in the bile fluid corresponded well with the dietary contamination. For all analyses it has to be noted that toxin residues were detectable even in individual samples of piglets fed the control diet containing 0.01 mg ZON/kg and 0.2 mg DON/kg. Presented at the 25^th Mykotoxin Workshop in Giessen, Germany, May 19–21, 2003     

Deoxynivalenol (DON) in raw and cooked Pasta

29 pasta samples were purchased from retail shops in Berlin in April and May 2001 and were analysed for their content of Deoxynivalenol (DON). Since it is much more important for the consumer to know the level of contamination with this mycotoxin in the ready to eat pasta we determined DON additionally in the cooked pasta and cooking water. 90% of the raw samples contained less than 0.5mg DON/kg, but three out of 29 samples up to 0.84mg/kg. The amount of DON decreased during cooking, which resulted in 60–80% lower DON levels in the ready to eat products. DON was analysed by HPLC-DAD, confirmatory analyses were carried out by GC-MS after derivatization of the analyte.     

An Investigation of the Soil Property Changes and Heavy Metal Accumulation in Relation to Long-term Wastewater Irrigation in the Semi-arid Region of Iran

Concentrations and spatial distribution of Zn, Cu, Cd, and Pb along two landscapes including a wastewater-irrigated area and a control area were determined to assess the impact of long-term wastewater irrigation and landscape properties on heavy metal contamination. Some disturbed and undisturbed soil samples were taken from soil trenches and soil cores, located on three main landscape positions (upper slope, midslope, and lower slope) in northwestern Iran. The investigation showed that the mean concentration of the heavy metals followed the order Zn > Cu > Pb > Cd in the wastewater-irrigated soil and Pb > Zn > Cu > Cd in the control soils. On average, compared to similar positions in the control region, the wastewater-irrigated regions contained 3.0 (midslope) to 4.9 (lower slope), 2.7 (midslope) to 4.6 (lower slope), 3.3 (upper slope) to 4.1 (lower slope), and 1.7 (upper slope) to 2.6 (lower slope) times higher amounts of Zn, Cu, Cd, and Pb, respectively. Significant positive relationships (P < 0.05) were recorded between the heavy metals concentration with <0.002 mm particle-size fraction and organic matter content, the fractions linked to runoff and soil erosion. It is believed that the two soil fractions play a crucial role in the distribution of the metals along the wastewater-irrigated landscape. Despite the significant increase of heavy metals (P < 0.05) in the wastewater-irrigated soils compared with control soils, the concentration of all evaluated metals was below the maximum accepted limits (Zn < 300 mg/kg, Cu < 100 mg/kg, Cd < 5 mg/kg, and Pb < 100 mg/kg), and grouped as “not-enriched” to “moderately-enriched” categories regarding the topsoil enrichment index. Overall, the lower slope was shown to be more contaminated with the heavy metals compared to the other positions.     

The potential effects of antioxidant feed additives in mitigating the adverse effects of corn naturally contaminated with <Emphasis Type="Italic">Fusarium</Emphasis> mycotoxins on antioxidant systems in the intestinal mucosa,plasma, and liver in weaned pigs

Seventy-two piglets (6.0 kg BW) were randomly distributed within six different dietary treatments to evaluate the effect of deoxynivalenol (DON) and the potential of four antioxidant feed additives in mitigating the adverse effects of DON on growth performances and oxidative status. Dietary treatments were as follows: control diet 0.8 mg/kg DON; contaminated diet (DON-contaminated diet) 3.1 mg/kg DON; and four contaminated diets, each supplemented with a different antioxidant feed additive, DON + vitamins, DON + organic selenium (Se)/glutathione (GSH), DON + quercetin, and DON + COMB (vitamins + Se/GSH + quercetin from the other treatments). Although DON was the main mycotoxin in the contaminated diet, this diet also contained 1.8 mg/kg of zearalenone (ZEN). The “mycotoxin” effects therefore included the combined effect of these two mycotoxins, DON, and ZEN. The DON-ZEN ingestion did not affect growth performances, average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency (G:F ratio), but partially induced oxidative stress in weaned pigs as shown by increased malondialdehyde (MDA) content in the plasma and superoxide dismutase (SOD) activity in liver (P?<?0.05). However, no change in the activity of other antioxidant enzymes or GSH concentrations was observed in plasma and liver of piglets fed the DON-contaminated diet (P?>?0.05). Supplementation with individual antioxidant feed additive had a limited effect in weaned pigs fed DON-ZEN-contaminated diets. Combination of antioxidants (vitamins A, C, and E, quercetin, and organic Se/GSH) reduced plasma and liver MDA content and SOD activity in liver (P?<?0.05) of piglets fed DON-ZEN-contaminated diets. Furthermore, this combination also reduced MDA content in the ileum (P?<?0.05), although activity of glutathione peroxidases (GPx), SOD or catalase (CAT) in the ileum was not affected by DON-ZEN contamination or antioxidant supplements. In conclusion, DON-ZEN contamination induced oxidative stress in weaned pigs and combination of antioxidant feed additives restored partially the oxidative status. Further studies will be necessary to assess whether the effects of antioxidant feed additives on oxidative status are specific when feed is contaminated with DON-ZEN.     

Investigation of the distribution of deoxynivalenol and ochratoxin A contamination within a 26 t truckload of wheat kernels

A study was conducted to determine the distribution of deoxynivalenol (DON) and ochratoxin A (OTA) in a lot of 261 of wheat kernels. Within this study, two different sampling and sample preparation strategies were carried out. On the one hand, following the official commission regulation 401/2006/EC, an aggregate sample out of 100 incremental samples was build, homogenized and prepared for laboratory analysis. On the other hand each individual subsample was investigated for its deoxynivalenol and ochratoxin A content. The determined concentration of DON in the individual samples was in a range from 830 up to 2655 (μg/kg, for OTA results ranged from < 0.2 up to 8.6 μg/kg. Thus, a coefficient of variance of 25% for DON and 200% for OTA was achieved. From this, a spot formation for OTA was observed and the average value of the 100 incremental samples did not correspond to the achieved value for the aggregate sample. While the DON contamination at this concentration range seems to be more even, consequently the result of the aggregate sample was in accordance with the average value. In addition a sample communition study was performed to answer the question whether the time consuming process of grinding of the whole aggregate sample is necessary or not. The results of this study show that contamination of whole wheat kernels with DON is at the same level within a 1 kg sample (CV 16%), while OTA contamination shows high variability (CV 94%). At least for OTA this study indicated that an extensive and complete sample communition of the high volume aggregate sample is necessary.     

On the effects of the <Emphasis Type="Italic">Fusarium</Emphasis> toxin deoxynivalenol (DON) administered per os or intraperitoneal infusion to sows during days 63 to 70 of gestation

Six pregnant sows of 180.6 ± 5.6 kg were fed either a Fusarium-contaminated (4.42 mg DON and 48.3 μg ZON per kg, DON per os, n = 3) or a control diet (0.15 mg DON and 5 μg ZON/kg) in the period of days 63 and 70 of gestation. On day 63 of gestation, sows fed the control diet were implanted with an intraperitoneal osmotic minipump (delivery rate of 10 μL/h, for 7 days) containing 50 mg pure (98%) DON in 2 ml 50% DMSO (DON ip, n = 3). Frequent plasma samples were taken to estimate the kinetics after oral and ip DON exposure. The intended continuous delivery of DON by the intraperitoneal minipump could not be shown, as there was a plasma peak (C_max) of 4.2–6.4 ng DON/mL either immediately (sow IP-2+3) or 2.5 h (sow IP-1) after implantation of the pump followed by a one-exponential decline with a mean half-time (t_1/2) of 1.75–4.0 h and only negligible DON plasma concentrations after 12 h. Therefore, the DON ip exposure has to be regarded as one single dose 1 week before termination of experiment. The DON per os sows showed a mean basis level (after achieving a steady state) of DON plasma concentration of about 6–8 ng/mL, as also indicated by the plasma DON concentration at the termination of the experiment. On day 70, caesarean section was carried out, the fetuses were killed immediately after birth, and samples of plasma, urine, and bile were taken to analyze the concentration of DON and its metabolite de-epoxy-DON. At necropsy there were no macroscopic lesions observed in any organ of either sows or piglets. Histopathological evaluation of sows liver and spleen revealed no alterations. The proliferation rate of peripheral blood mononuclear cells (PBMC) with or without stimulation was not affected by the kind of DON treatment. The exposure of pregnant sows at mid-gestation (days 63–70, period of organogenesis) to a Fusarium toxin-contaminated diet (4.42 mg DON and 0.048 mg ZON per kg) or pure DON via intraperitoneal osmotic minipump did not cause adverse effects on health, fertility, maintenance of pregnancy, and performance of sows and their fetuses. However, DON was detected in fetus plasma, indicating that this toxin can pass the placental barrier and may cause changes in the proportion of white blood cells (lower monocyte and neutrophil and higher lymphocyte proportion in DON per os fetuses).     

Assessment of Metal Availability in Soils from a Pb-Zn Mine Site of South-Central Spain

Heavy metal pollution of the soils around an abandoned Pb-Zn mine site located in the Alcudia Valley (South Central Spain) have been characterized by analysis of extractable and total metal concentrations in 60 samples of arable, pasture, and mine lands. The samples showed a broad range of size-particle distribution, cation exchange capacity, and pH values as well as high levels of total metal concentrations (up to 98510 mg kg^?1 of Pb, up to 20912 mg kg^?1 of Zn, and up to 61 mg kg^?1 of Cd). In order to assess the potential availability of metals the metal partitioning in two different soil size fractions (<2 mm and <63 μm) was determined using EDTA and CaCl₂ as sequestering reagents. The average contents of Pb, Zn, and Cd in the <63 μm particle size fraction for both extractions were higher than those of the <2 mm fraction due to the high metal adsorption capacity of the fine soil particles. Concentrations of heavy metals extracted by CaCl₂ were up to three orders of magnitude lower than those extracted by EDTA, because CaCl₂ only extracts the easily mobile fraction. Metal concentrations extracted by both procedures in the two granulometric fractions increased with total metal concentrations, thus increasing the potential environmental risk associated to heavy metal pollution.     

Urinary deoxynivalenol (DON) and zearalenone (ZEA) as biomarkers of DON and ZEA exposure of pigs

Four diets contaminated with 1.1 to 5.0 mg/kg deoxynivalenol (DON) and 0.4 to 2.4 mg/kg zearalenone (ZEA) were fed to four groups of six growing Large White pigs. Urine samples were collected after 3 to 4 days and again after 6 to 7 days on the diets. On each sampling day, half of the animals were sampled in the morning, after an 8-h fast, and the other half were sampled in the afternoon, after 7 h of ad libitum access to feed. The urinary concentrations of DON, DON-glucuronide, DON-3-sulphate, de-epoxy-DON, as well as of ZEA, ZEA-14-glucuronide, α-zearalenol and α-zearalenol-14-glucuronide, analysed using LC-MS/MS, were used to calculate urinary DON and ZEA equivalent concentrations (DONe and ZEAe). The urinary concentration of DONe (P?<?0.001), but not of ZEAe (P?=?0.31), was lower in the fasted than that in the fed animals. The urinary DONe/creatinine and ZEAe/creatinine ratios were highly correlated with DON and ZEA intake per kg body weight the day preceding sampling (r?=?0.76 and 0.77; P?<?0.001). The correlations between DON intake during the 7 h preceding urine sampling in the afternoon and urinary DONe/creatinine ratio (r?=?0.88) as well as between mean ZEA intake during 3 days preceding urine sampling and urinary ZEAe/creatinine ratio (r?=?0.84) were even higher, reflecting the plasma elimination half-time of several hours for DON and of more than 3 days for ZEA. ZEAe analysed in enzymatically hydrolysed urine using an ELISA kit was highly correlated with the LC-MS/MS data (r?=?0.94). The urinary DONe and ZEAe to creatinine ratios, analysed in pooled urine samples of several pigs fed the same diet, can be used to estimate their exposure to DON and ZEA.     

Field incidence of mycotoxins in commercial popcorn and potential environmental influences

Popcorn ear damage by insects and mycotoxin levels in kernels were monitored in several commercial popcorn fields in central Illinois over a 4-year period. Aflatoxin was rare, but fumonisin and deoxynivalenol (DON) were commonly encountered each year, and occurred at mean levels in fields up to 1.7 mg/kg (sample max. 2.77 mg/kg) and 1.9 mg/kg (sample max. 2.66 mg/kg), respectively. Neither fumonisin nor DON levels were significantly correlated with the percent of ears with visibly moldy insect-damaged kernels. Significant correlations were noted for the percent of ears with early caterpillar damage and both fumonisin and DON levels overall for some years and at specific sites in other years. Fumonisin levels were generally more highly correlated with insect damage than DON levels. Insect damaged kernels had 100- to 500-fold or greater levels of fumonisin compared to noninsect-damaged kernels, while DON levels were closer to 10- to 30-fold higher in insect damaged versus nondamaged kernels. A high percentage of DON-contaminated kernels were not insect damaged in 2007 and 2008. In some cases, differing mycotoxin levels for the same hybrid and same year planted at different locations appeared to be due to the prior crop. Higher DON levels in 2008 than other years were most likely associated with higher levels of rainfall and cooler temperatures than average during ear fill. While kernel sorters are reported to remove mycotoxin-contaminated popcorn kernels to acceptible levels, consideration of environmental factors that promote mycotoxins in popcorn should result in more effective control measures in the field.     

Preliminary exposure assessment of deoxynivalenol and patulin in South Africa

Deoxynivalenol (DON) and patulin (PAT) are mycotoxins widely regulated internationally. DON is frequently found in cereals, whereas PAT is commonly found in apple juices. A survey of South African commercial products was conducted on DON levels in maize meal and wheat flours, and on PAT levels in apple juices. DON levels in 23 wheat flour samples (mean of 16 positives, 29 μg/kg) were equal to or below 100 μg/kg and in wheat consumers contributed 6–13% of the provisional maximum tolerable daily intake (PMTDI; 1 μg/kg body weight per day) for DON set by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Sixteen of 18 maize meal samples were contaminated, with a mean for positive samples of 294 μg/kg, and the probable daily intakes ranged from 3.67 μg/kg body weight per day in rural infants to 1.39 μg/kg body weight per day in urban adults. PAT levels in 20 of 30 apple-juice samples were below the detection level of 10 μg/l. Mean of positive samples was 210 μg/l, with three samples exceeding the South African legal limit of 50 μg/l and the highest level (1,650 μg/l) showing the possibility of a brief but high exposure of 37 μg/kg body weight per day (or 9,250% of the JECFA PMTDI of 0.4 μg/kg body weight per day) in young children.     

Cumulation of mycotoxins in maize cobs infected with<Emphasis Type="Italic">Fusarium gramihearum</Emphasis>

Maize cobs withFusarium ear rot were collected at 1986 season and five infected byFusarium graminearum were analyzed for presence of triohothecenes and zearalenone. Collected material was subsampled forFusarium damaged kernels and corresponding axial stems and healthy looking kernels. All investigated cobs contained deoxynivalenol (DON) (range 18.0–131.5 mg/kg) and zearalenone (ZEA) (range 0.38–2.17 mg/kg), in four cobs 15-acetyl-deoxynivalenol (15-AcDON) (range 5.2–6.2 mg/kg) was present and two cobs besides three all metabolites contained 3-acetyl-deoxynivalenol (3-AcD0N) (range 0.5–0.8 mg/kg).The average of individual toxins amount in axial stems: in mg/kg was equal to: DON — 110.36, ZEA — 4.57, 15-AcD0N — 16.66, and 3-AcD0N — 1.32.Fusarium damaged kernels contained in average the following amount (mg/kg) of: DON 77.00, ZEA 0.98, 15-AcD0N 3.78 and 3-AcD0N 0.06. Healthy looking kernels contained DON 1.96 mg/kg and ZEA 0.07 mg/kg only. Cooccurrence of 3-AcDON and 15-AcDON in two samples was an interesting finding. The amount of DON in total cob was highly correlated (r = 0.94) with percentage ofFusarium damaged kernels in given ear.     

Occurrence of deoxynivalenol (DON) and ochratoxin A (OTA) in dog foods

The commercially available dog food samples (29 dry foods and 11 wet foods) were analysed for deoxynivalenol (DON) and ochratoxin A (OTA) using ELISA. All (100%) dry foods were contaminated with DON with various amount of the toxin (22-1837 μg/kg). In wet food 3 samples were found to be positive for DON in the range of 95-170 μg/kg. There were a few samples contaminated with OTA: 3 samples in dry foods (7-40 μg/kg) and 2 samples in wet foods (45 and 115 μg/kg).     

Sample clean-up methods,immunoaffinity chromatography and solid phase extraction,for determination of deoxynivalenol and deepoxy deoxynivalenol in swine serum

Concentrations of deoxynivalenol (DON) and deepoxy deoxynivalenol (DOM-1) in animal blood are important parameters for studies in toxicology and biological detoxification of DON. Clean-up methods, using either immunoaffinity chromatography (IAC) or solid phase extraction (SPE), were compared in order to determine the free form of DON or DOM-1 and the sum amount (free form plus glucuronide conjugated form of DON or DOM-1), respectively, in swine serum. Detection was achieved by high performance liquid chromatography with ultraviolet detection (HPLC-UV). Compared with the SPE-HPLC method, the IAC-HPLC method provided lower quantitation limit (DON: 18 vs 42 ng/ml; DOM-1: 21 vs 30 ng/ml) and higher recoveries (DON: 93.4–102.7% vs 63.7–85.3%; DOM-1: 85.5–91.1% vs 68.0–82.6%). Compared with previously published methods, the developed IAC-HPLC method removed analytical interferences from swine serum in one quick and easy step, and eliminated steps of extraction with organic solvent and/or pre-purification using SPE cartridges. This IAC-HPLC method was used to analyze swine serum samples collected from pigs that were evaluated in a feeding trial of a microbiological detoxification of DON. No DON or DOM-1 were detected in serum samples from pigs given a toxin-free diet or a microbial control diet. In serum samples from pigs given a DON diet (5 mg/kg of DON), free form DON and sum free DON + conjugated DON were 38.8 ± 13.7 and 49.8 ± 14.1 ng/ml, respectively. In serum samples from those given a detoxified-DON diet (DON was transformed to DOM-1), free form DOM-1 was detected but not quantified, and the sum DOM-1 was found as 47.5 ± 6.3 ng/ml.     

Survey and risk assessment of the mycotoxins deoxynivalenol,zearalenone, fumonisins,ochratoxin A,and aflatoxins in commercial dry dog food

The aim of the present study was to investigate the occurrence of mycotoxins in commercial dog food, as a basis to estimate the risk of adverse effects. Seventy-six dry dog food samples from 27 producers were purchased from retail shops, supermarkets, and specialized pet food shops in Vienna, Austria. The frequency and levels of deoxynivalenol (DON), zearalenone (ZEA), fumonisins (FUM), ochratoxin A (OTA). and aflatoxins (AF) in dry dog food were determined. Mycotoxin analysis were performed by commercial enzyme-linked immunosorbent assay (ELISA) test kits. Confirmatory analyses were done for DON, ZEA, and FUM by high performance liquid chromatography (HPLC) after extract clean-up with immunoaffinity columns. The correlations between ELISA and HPLC results for DON and ZEA were acceptable and indicated that ELISA could be a simple, low cost, and sensitive screening tool for mycotoxins detection, contributing to quality and safety of pet food. DON was the mycotoxin most frequently found (83% positives; median 308 μg/kg, maximum 1,390 μg/kg). ZEA (47% positives, median 51 μg/kg and maximum 298 μg/kg) and FUM (42% positives, median 122 μg/kg and maximum 568 μg/kg) were also frequently detected in dog food. OTA was less frequently found (5%, median 3.6 μg/kg, maximum 4.7 μg/kg. AF were not detected (<0.5 μg/kg) in any sample. The results show that dry dog food marketed in Vienna are frequently contaminated with mycotoxins (DON > ZEA > FUM > OTA) in low concentrations, but do not contain AF. The high frequency of Fusarium toxins DON, ZEA, and FUM indicates the need for intensive control measures to prevent mycotoxins in dog foods. The mycotoxin levels found in dry dog food are considered as safe in aspects of acute mycotoxicoses. However, repeated and long-time exposure of dogs to low levels of mycotoxins may pose a health risk.     

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.     

Analysis of theFusarium toxins Deoxynivalenol and Zearalenone in Austrian feeds of the crop years 2002 and 2003

164 feed samples were collected in Lower Austria, Upper Austria, Styria and Carynthia in the crop years 2002 and 2003 and were analysed by means of the RIDASCREEN^® ELISA-assay for theFusarium toxins Deoxynivalenol (DON) and Zearalenone (ZON). The collected samples consisted of swine feeds, cereals and silages. DON concentrations were found up to the range of mg/kg (ppm), in contrast ZON results remained significantly lower.     

Simultaneous occurrence of mycotoxins in human food commodities from Cameroon

Eighty-two samples of dried food commodities from Cameroon were screened and quantified for different mycotoxins, including fumonisin B₁ (FB₁), zearalenone (ZEA), deoxynivalenol (DON), aflatoxin (AF) and ochratoxin A (OTA), by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), respectively. The percentage of positive samples was as follows: FB₁ 41%, AF 51%, ZEA 57%, DON 65% and OTA 3%. High FB₁ contents were found in maize, averaging 3,684 μg/kg (range: 37-24,225 μg/kg), whereas the highest average ZEA level was found in peanuts (70 μg/kg), followed by maize (69 μg/kg), rice (67 μg/kg) and beans (48 μg/kg) with no ZEA was detected in soybeans. DON contents were low, ranging from 13 to 273 μg/kg, and for AF the average content was 2.6 μg/kg with peanuts and maize as principal substrates. The incidence of OTA was low, with a mean level of 6.4 μg/kg recorded. The majority (79%) of samples contained more than one mycotoxin and the most frequent co-occurrence found was FB₁ + ZEA + DON, detected in 21% of samples (mainly maize) analysed. Co-contamination with FB₁ + ZEA + DON + AF was found in 11% of the samples. Although a large proportion of samples had fairly low levels of individual mycotoxins, this should be of concern as the co-occurrence of mycotoxins may generate additive or synergistic effect in humans, especially if the respective commodities are consumed almost on a daily basis.