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.     

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.     

Co-occurrence of mycotoxins in swine feed produced in Portugal

A total of 404 samples of commercial swine feed from Portugal feed mills were analysed by HPLC methods for the presence of mycotoxins: 277 samples of feed for fattening pigs were analysed for ochratoxin A (OTA), zearalenone (ZEA), and deoxynivalenol (DON), and 127 samples of feed for sows were analysed for ZEA and fumonisins (FB₁ + FB₂). Concerning feed for fattening pigs, 21 (7.6%) samples were positive for OTA, (2–6.8 μg/kg), 69 (24.9%) were positive for ZEA (5–73 μg/kg), and 47 (16.9%) were positive for DON (100–864 μg/kg). In feed for sows, the results showed 29.9% of positive samples for ZEA (5–57.7 μg/kg) and 8.7% positive samples for FB₁ and FB₂ (50–391.4 μg/kg). Co-occurrence of DON/ZEA was found most frequently, but simultaneous contamination with OTA/ZEA and OTA/DON was also found.     

Occurrence of fumonisins in maize imported into Iran during 2001–2002

Fumonisins, fungal toxins found primarily in maize and produced by various Fusarium species, have been shown to cause a variety of significant adverse health effects in livestock and experimental animals, and are probable human carcinogens. Thirty-three maize samples were collected at ports from bulk shipments, which were imported into Iran from six countries during 2001–2002, and analysed by HPLC for the most abundant of the naturally occurring fumonisin analogues, namely fumonisins B₁ (FB₁), B₂ (FB₂) and B₃ (FB₃). Of the 33 samples, 21 (64%) were found to contain FB₁ (58–512 μg/kg) at levels above 10 μg/kg. The frequency of FB₁ found in maize samples imported from Uruguay and Canada was 75%, followed by China and Argentina (67%), USA (60%), and Brazil (50%). The average FB₁ level was 266 and 169 μg/kg for positive and all samples, respectively. Medians were 250 and 146 μg/kg for positive and all samples, respectively. FB₂ levels ranged from not detected (<10 μg/kg) to 53 μg/kg, whereas no sample had an FB₃ level above the detection level (10 μg/kg). This is the first report of fumonisin contamination of imported maize in Iran. Although, the level of all detected fumonisins were below the Iranian and FDA tolerance levels for foods and feeds, It is necessary to maintain the strict rules to ensure continued safety of imported maize.     

Occurrence of Fumonisin B<Subscript>1</Subscript> in Corn from the Main Corn-Producing Areas of China

Fumonisin B₁ (FB₁) is the most abundant of the fumonisin mycotoxins, mainly produced in maize by F. verticillioides and F. proliferatum. A total of 282 corn samples harvested in 2005 from six provinces, the main corn-producing areas of China, were analyzed for FB₁ using high-performance liquid chromatography. All samples except one were (99.6%) positive for FB₁ at levels varying from 3 to 71,121 ng/g with mean and median levels for all samples of 6,662 and 1,569 ng/g, respectively. During an analysis of the distribution pattern for FB₁, it became apparent that 43.6% of tested samples had FB₁ concentrations below 1,000 ng/g, while 25.2% contained in excess of 5,000 ng/g. The average exposure to FB₁ (1.1 μg/kg body weight/day) is within the provisional maximum tolerable daily intake of 2 μg/kg body weight/day set by the Joint FAO/WHO Expert Committee on Food Additives.     

Determinatıon of ochratoxin A and total aflatoxin levels in corn samples from Turkey by enzyme-linked immunosorbent assay

Forty-seven samples of corn were collected from various street bazaars and market outlets in different regions of Turkey and total aflatoxin (AF) and ochratoxin A (OTA) levels were determined by enzyme-linked immunosorbent assay (ELISA) following sample preparation. Levels of AF and OTA in corn samples ranged between 1.75–120.3 μg/kg and 1.08–8.57 μg/kg, respectively. Although 53% of the samples analysed had no detectable levels of AF, 4% of similar samples were found to contain AFs above the acceptable limit of 10 μg/kg in Turkey. For OTA, 4% of the corn samples had levels above the acceptable limit (3 μg/kg) in Turkey, with over 43% samples not found to contain this mycotoxin. Although the levels of mycotoxins analysed in this study were not found to be high and the percentage of samples contaminated above permitted limits were low, the importance of overall daily dietary intake should not be underestimated and control of these fungal metabolites in corn must be explored to minimise the hazards they may cause in humans.     

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.     

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.     

Content of the <Emphasis Type="Italic">Alternaria</Emphasis> mycotoxin tenuazonic acid in food commodities determined by a stable isotope dilution assay

The Alternaria mycotoxin tenuazonic acid (TA) was quantified in fruit juices (n = 50), cereals (n = 12) and spices (n = 38) using a recently developed stable isotope dilution assay (SIDA). [¹³ C₆,¹⁵ N]-TA was used as the internal standard. Method validation revealed low limits of detection (LODs) of 0.15 μg/kg (fruit juices), 1.0 μg/kg (cereals) and 17 μg/kg (spices). The respective limits of quantitation were about three times higher. Recovery was about 100% for all matrices. The precision (relative standard deviation of replicate analyses of naturally contaminated samples) was 4.2% (grape juice; 1.7 μg/kg), 3.5% (whole wheat flour; 36 μg/kg) and 0.9% (curry powder; 215 μg/kg). The median content of TA in the analyzed samples was 1.8 μg/kg (fruit juices), 16 μg/kg (cereals) and 500 μg/kg (spices). Positive samples amounted to 86% (fruit juices), 92% (cereals) and 87% (spices).     

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.     

Mycotoxins in several Polish food products in 2004–2005

In 2004–2005, samples of several selected Polish foods such as cereal products, nuts, dried fruits, coffee and culinary spices collected from Warsaw market and taken from food producers were analyzed on presence of aflatoxin B₁, B₂, G₁, G₂ (AF), ochratoxin A (OTA), zearalenone (ZEA) and deoxynivalenol (DON). After extraction and clean-up of extracts on immunoaffinity columns (IAC), mycotoxin analyses were carried out by HPLC using fluorescence and UV detectors. The concentrations of aflatoxins and ochratoxin A depending on the kind of sample ranged from 0.02 to 7.8 (one sample, of peanuts) and 0.02–11.9 μg/kg (one coffee sample), respectively. The levels of ZEA and DON were found to be below 50 °g/kg.     

Aflatoxins in peanut butter in Khartoum State,Sudan

Forty-three peanut butter samples from Khartoum State, Sudan, were analyzed for aflatoxins (AFs, AFB₁ + AFB₂ + AFG₁ + AFG₂) using high performance liquid chromatography (HPLC) with fluorescence detection after extraction with methanol:water (8:1, v/v) and clean-up using chloroform. All samples were contaminated with AFs, with total AF levels ranging between 26.7 and 853 μg/kg, and a mean total AF level of 287 ± 200.5 μg/kg. The highest concentrations were found for AFB₁, (28 positive samples, maximum 534 μg/kg), while AFG₁ was most frequently detected (43 positive samples, maximum 401 μg/kg). AFB₂ (42 positive samples, maximum 3.2 μg/kg) and AFG₂ (4 positive samples, maximum 30 μg/kg) were also present in these samples. The mean AF contamination levels found in this study exceeded by far all international regulations concerning maximum levels for this group of toxins. From the data, it is concluded that the levels of AF contamination in peanut butter from the Kartoum area are quite alarming, and may pose serious health hazards to consumers. Therefore, an intervention strategy to manage AF in peanut butter is urgently needed.     

A survey on the occurrence of mycotoxins in wheat and maize from western Romania

Samples of wheat (n = 25) and maize (n = 30) for animal consumption, collected in 1997 after harvest from western Romania, were analyzed by enzyme immunoassays for mycotoxin contamination. Toxins analyses included deoxynivalenol (DON), 3-acetylDON, 15- acetylDON, fusarenone X (FX), T-2 Toxin (T-2), diacetoxyscirpenol (DAS), zearalenone (ZEA), fumonisin B1 (FB1), aflatoxin B1 (AFB1), ochratoxin A (OA), and citrinin (CT). DON and acetylDONs were the major contaminants in wheat (100%) and maize (46%). Median values for DON, 3-acetylDON, and 15-acetylDON were 880 μg kg-1, 66 μg kg- 1, and 150 μg kg-1 in wheat, and 890 μg kg-1, 180 μg kg-1, and 620 μg kg- 1 in maize, respectively. Additionally, 3,15-diacetylDON was detected in some samples by HPLC-EIA analysis. All samples were negative for FX (<150 μg kg-1). T-2 was found in wheat (n = 6) and maize (n = 1) at levels between 13 and 63 μg kg- 1. DAS (2.6 μg kg-1) was found in one maize sample. ZEA occurred in all wheat and in four maize samples, median values were 10 μg kg-1 and 250 μg kg-1, respectively. One maize sample contained FB1 (140 μg kg-1). All samples were AFB1-negative (<4 μg kg-1). OA was found in one wheat sample (37 μg kg- 1), CT was found in one maize sample (580 μg kg- 1). This first reported natural occurrence of a range of mycotoxins in Romanian feeding stuff shows that DON and acetyl DONs may be present at levels which may affect animal production. This revised version was published online in June 2006 with corrections to the Cover Date.     

<Emphasis Type="Italic">Fusarium</Emphasis> toxins in cereals of integrated and organic cultivation from the Federal State of Brandenburg (Germany) harvested in the years 2000–2007

Over a period of 8 years (2000–2007), wheat (n = 407) and rye (n = 510) samples of integrated and organic cultivation in the Federal State of Brandenburg were analyzed by HPLC for the Fusarium toxins deoxynivalenol (DON) and zearalenone (ZEA). In the years 2002 and 2007, the overall contamination level was higher than in the other years. The percentage of DON-positives (>50 μg/kg) varied from 5 to 86%, the median and maximum levels varied from 50 to 380 μg/kg and from 50  to 10,400 μg/kg, respectively. The percentage of ZEA-positives (>3 μg/kg) varied from 2 to 41%, the median and maximum levels varied from 8 to 84 μg/kg and from 10 to 451 μg/kg. In the 8 years of testing, frequency and levels of DON and ZEA were significantly lower in cereals of organic cultivation compared with cereals of integrated cultivation.     

Determination of ergot alkaloids in feed by HPLC

A HPLC method for the determination of ergometrine, ergotamine, ergocristine, α-ergocryptine and ergocornine in cereals for animal feed and in mixed feed with high cereal content was developed. Samples were extracted under acidic conditions using a mixture of phosphoric acid and acetonitrile, the extract purified with solid phase extraction cartridges (strong cation exchange), and ergot alkaloids detected after gradient elution on a C18 column by HPLC with fluorescence detection. Detection and determination limits for each individual alkaloid were at 5 (μ/kg and 10 (μg/kg, respectively. With this method, high recovery (82–120%) and good reproducibility was achieved for wheat, rye and mixed feeds, at a sum of total determined alkaloids of < 500 (μg/kg. This method was used to analyse Bavarian feeds (n=124) over three years (2005–2007), and ergot alkaloids were detected in 91 % of the samples. The majority of positive samples had ergot alkaloid contents of < 250 μg/kg, the median alkaloid level was at 70 (μg/kg. The maximum sum of total determined alkaloids exceeded 1000 (μg/kg in wheat, triticale, rye, and mixed feeds, the highest result was obtained for mixed feed (4880 (μg/kg). Parts presented at the Feed Safety Conference, Namur, Belgium, Nov 27–28, 2007     

Survey of fungal counts and natural occurrence of aflatoxins in Malaysian starch-based foods

In a survey of starch-based foods sampled from retail outlets in Malaysia, fungal colonies were mostly detected in wheat flour (100%), followed by rice flour (74%), glutinous rice grains (72%), ordinary rice grains (60%), glutinous rice flour (48%) and corn flour (26%). All positive samples of ordinary rice and glutinous rice grains had total fungal counts below 103 cfu/g sample, while among the positive rice flour, glutinous rice flour and corn flour samples, the highest total fungal count was more than 103 but less than 104 cfu/g sample respectively. However, in wheat flour samples total fungal count ranged from 102 cfu/g sample to slightly more than 104 cfu/g sample. Aflatoxigenic colonies were mostly detected in wheat flour (20%), followed by ordinary rice grains (4%), glutinous rice grains (4%) and glutinous rice flour (2%). No aflatoxigenic colonies were isolated from rice flour and corn flour samples. Screening of aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2 using reversed-phase HPLC were carried out on 84 samples of ordinary rice grains and 83 samples of wheat flour. Two point four percent (2.4%) of ordinary rice grains were positive for aflatoxin G1 and 3.6% were positive for aflatoxin G2. All the positive samples were collected from private homes at concentrations ranging from 3.69–77.50 μg/kg. One point two percent (1.2%) of wheat flour samples were positive for aflatoxin B1 at a concentration of 25.62 μ};g/kg, 4.8% were positive for aflatoxin B2 at concentrations ranging from 11.25–252.50 μg/kg, 3.6% were positive for aflatoxin G1 at concentrations ranging from 25.00–289.38 μg/kg and 13.25% were positive for aflatoxin G2 at concentrations ranging from 16.25–436.25 μg/kg. Similarly, positive wheat flour samples were mostly collected from private homes. This revised version was published online in June 2006 with corrections to the Cover Date.     

Airborne mycotoxins in dust from grain elevators

Workers in grain elevators are exposed to grain dust and may therefore have an increased risk of inhalatory contact with mycotoxins. To study the mycotoxin burden of such environments, settled grain dust samples (n=35) were collected from several locations of a total of 13 grain elevators in Germany, and analysed for ochratoxin A (OTA, detection limit 0.01 ng/g), deoxynivalenol (DON, detection limit 15 ng/g), and zearalenone (ZEA, detection limit 6 ng/g), respectively. Cytotoxicity of these samples was assessed by a MTT bioassay with a swine kidney target cell line. Additionally, the airborne dust concentration of these locations was determined. Nearly all settled dust samples contained OTA (96%), DON (100%), and ZEA (100%) with median concentrations of 0.4 ng/g, 416 ng/g, and 126 ng/g, respectively. Cytotoxic effects in varying degrees from weakly to highly toxic were caused by crude extracts of 86% of the dust samples. However, cytotoxicity did not correlate with mycotoxin levels in these samples and thus indicated the presence of cytotoxic compounds of unknown origin. Based on the mycotoxin findings in settled dust samples and the airborne dust concentrations, the average airborne mycotoxin concentrations were estimated to be 0.002 ng/m³ (OTA), 2 ng/m³ (DON), and 1 ng/m³ (ZEA), respectively. The relevance of these findings for occupational health was assessed by comparison with WHO recommendations for the maximum tolerable daily (oral) intake (TDI). Even in a worst case scenario, the calculated inhalatory intake was far below the TDI values. However, considering the uncertainties resulting from different exposure pathways, namely oral ingestion versus inhalation, further research should primarily address the problem of how adequate assessment criteria for airborne exposure to mycotoxins could be established. Presented at the 28^th Mykotoxin-Workshop, Bydgoszcz, Poland, May 29–31, 2006     

Fusarium toxin contents of maize and maize products purchased in the years 2000 and 2001 in Germany

Samples (n=106) of maize and maize products were analysed for 13 trichothecene toxins and zearalenone (ZON). All 14 toxins examined were detected, although with varying frequency. Cooccurrence of two or more toxins was observed in 96% of samples. The toxins of the scirpenol group scirpentriol, 15-monoacetoxyscirpenol and diacetoxyscirpenol were detected in 14, 27 and 3% of the samples analysed, the toxins of the T-2 group T-2 toxin, HT-2 toxin, T-2 triol und T-2 tetraol were found in 33, 66, 2 and 7%. Toxin content was higher in feeds than in foods (semolina and flour). In food samples, the German regulatory level for DON (500 μg/kg) was not exceeded, three samples of maize flour contained ZON above the regulatory level (50 μg/kg). Presented at the 26^th Mykotoxin-Workshop in Herrching, Germany, May17–19, 2004     

Occurrence ofAlternaria andFusarium mycotoxins in winter wheat from domestic crop in year 2003

The aim of this study was a monitoring of the occurrence ofAlternaria andFusarium mycotoxins in winter wheat from domestic crop in the year 2003. Altenuene was determined in 56 (100%) samples of winter wheat, range 14.5–41 μg/kg, mean 25 μg/kg. Alternariol was determined in 16 (28.6%) samples of winter wheat, range 6.3–22.1 μg/kg, mean 5.7 μ/kg. DON was determined in 42 (100%) samples of winter wheat, range 250–3500 μg/kg, mean 330 μg/kg. T2-toxin was determined in 42 (100%) samples of winter wheat, range 25–337 μg/kg, mean 99 μg/kg. ZEA was not determined in samples of winter wheat. Presented at the 26^th Mykotoxin-Workshop in Herrsching, Germary, May 17–19, 2004 Financial support. Supported (one part of experiments, the determination of Fusarium mycotoxins) by the Ministry of Agricu ture of the Czech Rebublic (Propect No QF3121)