Relationship between Fusarium graminearum and Alternaria alternata contamination and deoxynivalenol occurrence on Argentinian durum wheat

A mycological survey was carried out on durum wheat (Triticum durum) samples from the main production area of Argentina. The isolation frequency and relative density of species of dematiaceous fungi, and genus Fusarium were calculated. Alternaria alternata and Fusarium graminearum were the predominant fungal species. An analysis of deoxynivalenol (DON) natural contamination was also performed on a limited number of samples (60). DON contamination levels in positive samples ranged from 26 to 6400 μg/kg. The non-parametric techniques applied showed that there is a positive relationship between DON contamination and F. graminearum relative densities and a negative relationship between DON contamination and A. alternata relative densities. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

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.     

Deoxynivalenol in lebensmitteln: Ergebnisse einer Pilotstudie

Cereal food products (n=333) were purchased in retail stores from Germany in 2001 and analysed for deoxynivalenol (DON), either by enzyme immunoassay or by HPLC after immunoaffinity chromatographic cleanup. Detection limits were dependent of the sample matrix and varied from 20–100 μg/kg. The overall DON incidence was 53%, with mean and median levels for positives of 251 μg/kg and 142 μg/kg, respectively. The contamination with DON (mean/median value, μg/kg) as found for bread (90/87), wheat flour (161/124), and noodles (472/297) indicate that the levels of DON in cereal foods were significant in view of the tolerable daily intake (1 μg/kg body weight) as established by the European Union scientific committee on food.     

Differences Between Barley, Oats and Wheat in the Occurrence of Deoxynivalenol and Other Trichothecenes in Norwegian Grain

The amount of the Fusarium toxins deoxynivalenol (DON). 3-acety-DON. ivalenol and fusarenon × have been determined in about 3000 samples of barley, oats and wheat grown in Norway. The samples were collected in the period 1988–1994. and grouped according to grain species, year of production, district and sample category. The DON content was found to be significantly higher in oats than both barley and wheat in grain produced by ordinary grain producers in Norway. Of samples collected from grain silos. 68.7% of the barley. 97.1% of the oats and 66.3% of the wheat samples were found to contain 30 mg/kg of DON or more. The proportion containing 1000 μg kg or more was 2.1.13.4and0.0% in burley, oats and wheat, respectively. However, these variations in contamination level were not reflected in the results from two field trials, where the grain species were grown side-by-side in the same fields. The differences in contamination level between the grain species seem to be due partly to edaphic and agrotechnical factors and partly to variation in the susceptibility to different Fusarium species in interaction with climatic factors. Nivalenol(≥ 50 μg/kg) was detected in 8.3% of the barley samples. 8.2% of the oats and 0.4% of the wheat samples. Samples collected at the grain silos were then not included.     

Mycotoxins in foods in Lower Saxony (Germany): results of official control analyses performed in 2009

In this presentation, the mycotoxin levels—as analysed by the analytical centre for mycotoxin surveillance of the state food laboratory (LAVES Braunschweig)—for approximately 500 food samples are reported. The samples were collected in the year 2009 at retail in the German federal state of Lower Saxony. Aflatoxin and ochratoxin A were analysed in dried fruits, spices, cereals and tree nuts. Ochratoxin A was detected in all samples of dried vine fruits, at levels up to 8.1 μg/kg. Aflatoxins and ochratoxin A were also found in nutmeg and curry powder: the maximum regulatory levels for aflatoxins were exceeded in 25% of the nutmeg samples. Nearly all samples of basmati rice contained aflatoxins, although at levels below the maximum regulatory level in all but one sample. Aflatoxins were also detected in about 50% of hazelnut samples, in 20% of the samples the maximum levels was exceeded (maximum 23.2 μg/kg). In contrast, aflatoxin contents in pistachios were surprisingly low. Fusarium toxins were analysed in cereals and cereal products such as flour, bread, and pasta. Deoxynivalenol (DON) was the predominant toxin found in these samples: DON was found in about 40% of the samples, although the maximum levels were not exceeded (max. 418 μg/kg). Fumonisins (FBs) and zearalenone (ZEA) were specifically analysed in maize products (snacks, flour and oil). Most of these samples (80%) were positive, but at levels not exceeding the maximum levels. Maximum levels were 98 μg/kg (ZEA) and 577 μg/kg (sum of FB₁ and FB₂). Ergot alkaloids (six major alkaloids) were analysed in rye flour, and approximately 50% were positive. The highest concentration of ergot alkaloids was 1,063 μg/kg; the predominant alkaloids were ergotamine and ergocristine. In conclusion, the results indicate that continuous and efficient control measures for mycotoxins in a wide range of critical foods are necessary to ensure compliance with maximum levels. Although the mycotoxin levels in the vast majority of samples were below maximum levels, year-to-year variation and changes in the production of relevant commodities may result in a different picture in the future.     

Zearalenone, deoxynivalenol and fumonisins in mixed-feed for laying hens

Fusarium toxins are secondary metabolites produced byfungi of these genera in many commodities under certain conditions. A study was carried out to investigate the co-occurrence of zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FB₁ and FB₂) in 52 samples of mixed-feed for poultry contaminated withFusarium verticillioides. The zearalenone and deoxynivalenol were checked using immunoaffinity column and the extraction of fumonisin was performed by strong anion exchange (SAX) solid phase column. Detection and quantification were determined by high performance liquid chromatography (HPLC). The limit of detection was 5 μg/kg for ZEN, 100 μg/kg for DON and 50 and 100 μg/kg for FB₁ and FB₂ respectively.Fusarium toxins were detected in 20 samples. Sixteen samples were positive for ZEN (30.7%) presenting levels that ranged from 7.4 μg/kg to 61.4 μg/kg (mean=27.0 μg/kg). 13.5% of the samples presented contaminations of DON, with levels ranging from 100.0 μg/kg to 253 μg/kg (mean=l18.07 μg/kg). FB₁ was detected in 19.2% of samples, with levels ranging from 50.0 μg/kg to 110.0 μg/kg (mean=73.6 μg/kg). FB2 was not detected in any sample. In positive samples simultaneously contamination with two or three mycotoxins were detected in 9 of them (17.3%).     

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.     

Deoxynivalenol in Mehlproben des Jahres 1999 aus dem Einzelhandel

To study the levels of deoxynivalenol (DON) in retail cereal products, wheat and rye samples were purchased in 1999 from supermarkets and “organic food” shops in Munich, Germany. DON was analysed by an enzyme immunoassay (EIA), 78 of these samples were additionally analysed by HPLC. The following contamination rates (%) and mean DON levels were found: wheat flour type 405 (n=42): 71%, 200 µg/kg; flour type 550 (n=9): 33%, 410 µg/kg; flour type 1050 (n=11): 91%, 370 µg/kg; bread-baking wheat premixes (n=14): 79%, 210 µg/kg; whole grain flour (n=20): 65%, 300 µg/kg; whole grain wheat (n=8): 75%, 280 µg/kg, wheat bran (n=20): 85%, 830 µg/kg; rye flour and grits (n=7): 29%, 120 µg/kg. HPLC confirmed the results obtained by EIA. Further analysis of 16 wheat flour (405) samples in May 2000 showed a similar frequency (69%) and mean DON level (270 µg/kg) as for samples from 1999. It is concluded that with DON levels in wheat for human consumption ranging from 200–400 µg/kg, the intake of DON has to be taken seriously in the light of the temporary tolerable daily intake of 1 µg DON per kg body weight as proposed within the European Union.     

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.     

<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.     

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)     

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).     

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.     

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     

Untersuchungen zu DON-Gehalten in Getreideerzeugnissen aus dem deutschen Einzelhandel im Jahr 2006

Samples of soft wheat flour (n=78), durum wheat semolina (n=6), and pasta (made from durum wheat, n=49) were purchased in January-April 2006 from retail outlets in Hesse, Germany. Samples were analysed for deoxynivalenol (DON) by enzyme immunoassay. The detection limit of the method was 10 μg/kg, with recoveries of 81–85% (RSD_r: 12–17%). DON was detected in 84% of all samples, but the contamination level was low. Median/maximum values for DON in wheat flour, wheat semolina, and pasta were 28μg/kg/217 μg/kg, 38μg/kg/203 μg/kg, and 24μg/kg/119 μg/kg, respectively. Compared with results obtained from previous years, significantly lower DON levels were observed in these commodities.     

Natural occurrence of zeralenone in feeds and feedstuffs used in poultry and pig nutrition in colombia

A total of 200 samples of feedstuffs and mixed feeds used for poultry and pig nutrition in Colombia were analysed for zearalenone using a LC technique with a limit of detection of 20 μg/kg. Samples of grain sorghum, maize, processed soybean, rice meal, cottonseed meal, and poultry and pig feeds, representative of the Colombian production for the 1995–1996 harvest, were taken at feed manufacturing plants located in different cities of the country. Zearalenone was detected in 25 of 45 samples of sorghum, 2 of 33 samples of maize, 7 of 22 samples of rice meal, 9 of 17 samples of cottonseed meal, 11 of 30 samples of poultry feed and 6 of 16 samples of pig feed. Zearalenone was not detected in soybean or other feedstuff s analysed (wheat by- products, cassava meal, palm). Overall levels of zearalenone ranged from 29 to 3956 μg/kg, with a mean value of 436 μg/kg. Only one of the 6 positive samples of pig feed had a zearalenone concentration above 500 μg/kg, which is normally considered as the safe level for pigs.     

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.     

The Effect of Nitrogen Fertilization on Fusarium Head Blight Development and Deoxynivalenol Contamination in Wheat

The impact of nitrogen (N) fertilization on the development of Fusarium head blight (FHB) in wheat and the resulting deoxynivalenol (DON) contamination in the kernels was studied. In a first experiment, the disease was assessed on two locations under natural infection pressure. Five different types of nitrogen fertilizer (both organic and mineral) were investigated, each applied at five input rates from 0 to 160 kg N/ha. With all fertilizers, a significant increase of disease intensity was observed with increasing N input, while the type of N fertilizer had poor or no effects on FHB. Depending on the fertilizer used, the percentage of diseased spikelets increased from 2.2% at zero N rate up to 6.6% at 160 kg N input per hectare. In a second series of trials, three spring wheat varieties including one Durum wheat line were artificially inoculated with a Fusarium graminearum and a F. culmorum strain, known producers of DON. A mineral N fertilizer was applied at five input levels from 0 to 160 kg N/ha. A significant increase in FHB intensity and DON contamination in the grain was observed with increasing N from 0 to 80 kg/ha. At higher input rates, relevant in contemporary crop husbandry, disease intensity and toxin contamination remained at constant levels. It is concluded that adaptation of N fertilization represents no relevant tool in managing FHB in practical wheat cultivation.     

First survey on the natural occurrence of Fusarium mycotoxins in Bulgarian wheat

Wheat for human consumption (140 samples) was collected after harvest from all regions of Bulgaria. The 1995 crop year was characterized by heavy rainfall in the spring and summer months. The internal mycoflora of wheat samples was dominated by Fusarium spp. and Alternaria spp., and storage fungi were rarely present. The samples were analysed for contamination with Fusarium mycotoxins deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON), T-2 Toxin (T-2), diacetoxyscirpenol (DAS), and zearalenone (ZEA), using enzyme immunoassay methods. DON and ZEA were the predominant toxins, with a contamination frequency of 67% and 69%, respectively. The average levels of these toxins in positive samples were 180 g/kg (DON) and 17 g/kg (ZEA), maximum concentrations were 1800 g kg^–1 and 120 g kg^–1, respectively. Acetyl derivatives of DON, namely 3-AcDON and 15-AcDON, were found in 2.1 % and 0.7% of the samples, at at maximum level of about 100 g kg^–1. Only one sample was positive for T-2 (55 g/kg), DAS was not detected. This is the first report about the natural occurrence of a range of Fusarium mycotoxins in wheat for human consumption in Bulgaria.Abbreviations 3-AcDON 3-acetyldeoxynivalenol - 15-AcDON 15-acetyldeoxynivalenol - DAS diacetoxyscirpenol - DON deoxynivalenol - EIA enzyme immunoassay - T-2 T-2 toxin - ZEA zearalenone