Natural Occurrence of Aflatoxins from Maize in Iran

Fifty-one maize samples, intended for animal feed and human consumption, were collected from the four main maize production provinces in Iran and analyzed by high-performance liquid chromatography (HPLC) for contamination by four naturally occurring aflatoxin analogues (AFB1, AFB2, AFG1, and AFG2). AFB1 was detected in 58.3, and 80% of the maize samples obtained from Kermanshah and Mazandaran provinces, respectively. The maximum AFB1 (276.3 μg/kg) and highest level of total aflatoxins (AFT) (316.9 μg/kg) were detected in a maize sample collected from Kermanshah province. The mean aflatoxin level from contaminated samples (52.60 μg/kg) from Kermanshah was significantly higher (P < 0.0001) than those in maize from the other three provinces and exceeded all the maximum tolerated levels (MTLs) set for AFT in maize. The level of AFB1 in 15.68% of the total samples was above the MTL (5 μg/kg) for AFB1 in maize in Iran. The mean contamination level of AFT (23.86 μg/kg) in the positive samples was higher than MTL for maize in Iran (20 μg/kg) intended for animal feed. The levels of AFB1, AFB2, AFG1, and AFG2 ranged between not detected (<0.1 μg/kg) and 276.3, 30.4, 9.1, and 1.1 μg/kg in maize grain, respectively.     

A survey of fumonisins (B1, B2, B3) in Indonesian corn-based food and feed samples

In this paper a survey is described for determination of contamination level of fumonisins (B¹, B², B³) in Indonesian cornbased feed and food samples. The survey was conducted from February to May 2001. Foodstuffs, which are consumed directly such as snacks and other products, were investigated for fumonisin contamination. Of 105 food and feed samples purchased from local retail stores and local poultry shops around Yogyakarta, Java, Indonesia were analyzed using ELISA. Results indicate that 74.3% of samples analyzed were contaminated in a large range of 10.0 – 3307 μg/kg, and the concentration of fumonisins depends on the type of samples. Detection limit of the method used was 9 μg/kg.From eight food samples of maize flour, and corn-based beverages and cereals, none was contaminated (below detection limit). For food samples of industrial products (19 samples), 13 were contaminated in the range of 22.8 – 105 μg/kg and 19 of 20 samples from home made products were contaminated between 12.9 – 234 μg/kg. The food samples contaminated in highest level occurred in corn. Of ten samples, 6 were contaminated from 68.0 – 2471 μg/kg. For feed samples, 17 corn samples were evaluated. Of those samples, 16 contained in a large range of 17.6 – 3306 μg/kg.     

Natural occurrence of ochratoxin A and citrinin in food stuffs in Egypt

Natural occurrence of ochratoxin A (OA) and citrinin in cereals (274 samples) and animal tissues (250 samples) have been investigated during a period of more than 2 years. OA was found in cereals and animal tissues while citrinin was found in cereals only. The highest level of OA (up to 80.0 μg/kg) was found in yellow corn, 52.8% of contaminated samples while respectively 55.9% and 39.4% of barley and rice samples were contaminated with citrinin, with the highest level up to 100.0 and 27.92 μg/kg for barley and rice respectively. The frequent contamination of animal kidney with OA (28% positive out of 150 tested) average concentration 12.33 μg/kg. 2% of liver and 4% of muscles tissue were observed.     

Occurrence of fumonisins (B<Subscript>1</Subscript>, B<Subscript>2</Subscript>, B<Subscript>3</Subscript>) in maize-based food and feed samples from Indonesia

A survey to evaluate the contamination level of total fumonisins in maize-based foodstuffs, maize and feed from Indonesia is described. The analyses were carried out by enzyme-linked immunosorbent assay (ELISA). Samples were collected from local retail stores around Yogyakarta, Indonesia between February and May 2001. The 101 samples were classified into six categories, i.e. industrially-produced food (n=24), products of small food manufacturers (n=17), maize flour (n=4), maize for food (n=9), maize for feed (n17), and formulated feed (n30). Control of the method showed that the detection limit was 8.7 μg/kg and repeatability is shown by relative standard deviation (RSD) of analyses of contaminated maize (n=5) of 10 %. Results of analyses indicate that 80 samples analysed were contaminated over a large range from 10.0-3307 pg/kg, and the concentration of fumonisins depended on the type of sample. Of four samples of maize flour, none were contaminated (below detection limit). Of 24 samples of industrially produced food, 14 were contaminated in the range 22.8 - 105 μg/kg and 18 of 19 food samples from small manufacturers were contaminated ranging from 12.9 to 234 μg/kg. The highest contamination was observed in maize samples: six of ten samples of maize for food were contaminated between 68.0 - 2471 μg/kg and 16 of 17 samples for feed contained fumonisins over a large range from 17.6 to 3306 μg/kg.     

Fusarium toxins in cereals — Results from eight German Federals States in 2000

A total of 353 cereal samples from 8 German Federal States were analysed for Fusarium toxins in the 2000' survey. The level of mycotoxin contamination of the samples was higher than in the previous year due to higher moisture during flowering in some regions. Concerning deoxynivalenol (DON), the prevailing mycotoxin in all the samples, we have to consider, that 24% of the samples as well as the average concentration exceed the advisory level of 500 μg/kg. The application of Fusarium active fungicides reduced the DON level from 963 μg/kg (samples without treatment) to 630 μg/kg (all samples from treated and untreated fields). It has to be emphasised that the number of nivalenol (NIV) positive samples increased in comparison to the previous year from 9% to 17%. In addition, an increase in the level of NIV was detected, the maximum concentration in winter wheat was 3480 μg/kg. Cereals were rather moderately contaminated with zearalenone (ZEA), more than 80% of the samples contained less than 50 μg/kg.     

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.     

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.     

Zeralenone and ergosterol contents in corn samples of Transylvania, Romania

70 corn samples for feed use out of the harvests of the years 2001 and 2002 were collected randomly in Transylvania, Romania and analysed for zearalenone and for ergosterol as fungal mass indicator. High ergosterol contents were found in some samples with a maximum value of 72 mg/kg, zearalenone contents showed values between 4 and 2250 μg/kg in samples positive, indicating a rather high degree of fungal contamination as well as a serious zearalenone contamination. Presented at the 25^th Mykotoxin Workshop in Giessen, Germany, May 19–21, 2003     

Analysis of T-2 and HT-2 toxins in oats and other cereals by means of HPLC with fluorescence detection

Bearing in mind the high toxicity of T-2 and HT-2 toxins which occur in cereals (mainly in oats) EU plans legal limits for these mycotoxins. The occurrence data are insufficient because reliable and sensitive analysis methods are not available. A sensitive HPLC gradient method was developed which is applicable with common HPLC equipment (HPLC with fluorescence detection). After extraction of the toxins from sample matrix with methanol/water the diluted extracts were cleaned-up using immunoaffinity columns and then derivatized with 1-anthroylnitrile/DMAP. The T-2 and HT-2 toxins were separated from peaks of the cereal matrix and derivatization reagent by means of a relatively complex HPLC gradient method. The method was validated for oats, wheat, rye, barley, and maize. The recovery rates were in the range of 70–99%, the precision (RSD_R) of 3–8%. The limits of detection of T-2 and HT-2 toxins were 1 μg/kg. A total of 119 samples of cereals and cereal products was analyzed according to the optimized method. The analyses of 54 samples of dehulled oats and of 11 samples of processed oat products from food industry had a contamination frequency of 100%. The contents (sum of T-2 and HT-2 toxins) amounted to 3 to 174 μg/kg for the dehulled oats and to 4 to 48 μg/kg for the processed oat products. 29 samples of maize and maize products had a contamination frequency of 80% (2–106 μg/kg in the sum of T-2 and HT-2 toxins). In the samples of wheat and barley the toxins were detected only occasionally (contents: 1–10 μg/kg), in rye not at all.     

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.     

Natural occurrence of aflatoxin B<Subscript>1</Subscript> in some Indonesian food and feed products in Yogyakarta in year 1998–1999

A survey was conducted between 1998–1999 to evaluate the level of aflatoxin B₁ (AfB₁) contamination in some selected Indonesian food products, mainly peanuts and peanut products for sale in supermarkets or traditional markets in Yogyakarta, Indonesia. Quantitative analysis was carried out on 118 samples using the ELISA (Enzyme-Linked Immunosorbent Assay) technique. The results indicate that (61.1%) samples were contaminated with AfB₁ at range 2.0 to 249.0 μg/kg. Approximately 50% of the baby food products analysed were contaminated with AfB₁ and the maximum level found was 7.0 μg/kg. In corn products and fermented products, AfB₁ was detected in 66.7 and 50.0% of samples, respectively. A level as high as 5.6 μg/kg of AfB₁ was found in the corn and 6.0 μg/kg in fermented product. AfB₁ was also detected in all rice products, feed products, and other processed products at levels of up to 7.0, 27.0, and 26.0 μg/kg, respectively.     

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.     

Natural occurrence of aflatoxins,deoxynivalenol, fumonisins and zearalenone in maize from Entre Ríos Province,Argentina

In Entre Ríos, Argentina, corn is one of the most important cereal grains produced, being an important income for the regional economy. The aim of this work was to assess aflatoxins, zearalenone, deoxynivalenol (DON) and fumonisins (FB) in corn harvest in 2003 and 2004 in the most contaminated departments found in previous studies in selected sampling places. At the harvest time, when the trucks arrived to store plants, samples of corn were taken from seven different positions of the trucks and from five in the trailer. Composite samples were randomised reduced to 10 kg. The samples were analysed by immunological tests, by thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and/or gas liquid chromatography-electron capture detector (GLC-ECD). In 2003 average contamination was 3.19 u.g/kg for aflatoxins, 118.5 μg/kg for deoxynivalenol, 230.8 μg/kg for zearalenone and 10200 μg/kg of total fumonisins (HPLC and ELISA quantification showed a linear correlation (r² =0.9618), but RIDASCREEN®FAST values were 1.7 higher than HPLC values); in 2004 deoxynivalenol and zearalenone were not detected and an average of 2.0 μg/kg for aflatoxins and 4700 μg/kg for total fumonisins was found.This province, with the earliest harvested corn in the country each summer, tends to display different contaminations from the rest of the provinces, probably due to climate characteristics, particularly hotter weather.     

Vorkommen und Nachweis von Fusarinsäure in Weizen

An analytical method for the detection of fusaric acid in wheat and wheat products has been developed as well as employed for the survey of randomly selected samples (n=33). While the application of GC-FID proved to be not sensitive enough for this purpose, the sensitivity and selectivity of GC-MS permitted the quantification of the mycotoxin in 78% of the wheat samples.The measured concentrations ranged from 7 to 570 μg/kg (average value 134 μg/kg). However, clear differences were recognizable regarding the type of sample: While field samples showed 332 μg/kg on an average, cleaned grains designated for human consumption contained averaged 138 μg/kg, and wholemeal products 68 μg/kg.Although the limited number of samples does not permit final conclusions, the toxicological relevance of fusaric acid alone may probably be classified as small. To what extent the well known cumulative effects with deoxynivalenol and fumonisin B1 impair this evaluation has to remain elusive.     

Metabolisierung von Deoxynivalenol beim Schwein: Bestimmung von DON und DOM-1 im Urin vom Schwein

This paper describes a feeding study with 7 pigs, which were fed with deoxynivalenol contaminated oats at a level of 0.23 mg/kg body mass/day over 16 experiment days. The contamination level of consumed feed was 14.4 mg DON/kg in the ration. The parallel control group of 7 pigs were fed with DON free oats. Urine samples were taken each two days. The content of DON and DOM-1 (de-epoxy deoxynivalenol) in urine was determined. The mean concentration of DON in urine of animals in trial group was 580 μg/l, whereas DOM-1 32 μg/l.     

Risk Assessment for Natural Uranium in Subsurface Water of Punjab State,India

Traces of uranium were measured by laser fluorimeter in 235 subsurface water samples collected from four districts of Punjab state in India. The concentration of U in water samples ranged between <2–644 μg/L with a mean value of 73.1 μg/L. The radiological risk was observed to be in the range of 5.55 × 10^?6–1.78 × 10^?3 with a mean value of 2.03 × 10^?4, which is around 22% more than the maximum acceptable level (l.67 × 10^?4) as per guidelines of India's Atomic Energy Regulatory Board. The mean of chemical toxicity risk, expressed as life time average daily dose (LADD) was worked out to be 5.56 μg/kg/day with a range of 0.15–48 μg/kg/day by considering a bodyweight of 51.5 ± 8.5 kg, water ingestion rate of 4.05 L/d, and life expectancy of 63.7 yrs for an adult Indian reference man and compared with the reference dose (4.53 μg/kg/day). The average exposure level of U was comparatively high and the chemical toxicity was expected to be more. The mean of hazard quotient (LADD/ RfD) for all four districts was found to be greater than 1, indicating that groundwater may not be suitable for consumption from a chemical toxicity point of view.     

Nachweis von Zearalenon-4-β-D-Glucopyranosid in Weizen

Maize (Zea mays) cell cultures were used for the production of zearalenone-4-β-D-glucopyranoside as standard compound. Wheat samples were extracted with acetonitrile: water, applied to a florisil column and eluted with methanol:ethyl acetate. For determination and quantification of zearalenone-4-β-D-glucopyranoside and zearalenone a LC-MS method was developed. A concentration of 10 μg/kg zearalenone-4-β-D-glucopyranoside and zearalenone was detectable. The recovery rates were calculated to be 69% and 89% at a concentration level of 100 μg/kg for zearalenone-4-β-D-glucopyranoside and zearalenone, respectively.24 Bavarian wheat samples from harvest 1999 were analyzed. Zearalenone was present in 22 out of 24 field samples, the levels ranged from 11–860 μg/kg. Zearalenone-4-β-D-glucopyranoside was found in 10 out of the zearalenone positive samples (42%) at levels ranging from 17 to 104 μg/kg. The amounts of zearalenone-4-β-D-glucopyranoside were correlated to those of zearalenone (r2=0,86; b=0,10).     

Comparing aflatoxin contamination in chilies from Punjab,Pakistan produced in summer and winter

A comparison was made of total aflatoxins (AFs) in 43 samples of chilies collected during winter and 42 in summer to determine the effect of season on contamination. The samples were analyzed by HPLC with fluorescence detection. The limits of detection and quantification for AFB₁ and AFG₁ were 0.05 μg/kg and 0.50 μg/kg, whilst for AFG₂ and AFB₂ they were 0.10 μg/kg and 0.60 μg/kg. In the winter samples, AFs were detected in 18 (72%) whole and 14 (60%) ground chilies, with concentration ranges 0.00-52.30 μg/kg and 0.00-74.60 μg/kg respectively. In the summer samples, 17 (64%) whole and 12 (76%) ground chilies were contaminated with AFs at concentrations 0.00-61.50 μg/kg and 0.00-95.90 μg/kg respectively. The percentage of samples greater than the European Union statutory limit for AFB₁ and total AF for whole chilies were 48 and 36%, compared with ground chili values of 50 and 45%, respectively, in the winter season. In the summer season, the samples greater than the European Union limit for AFB₁ and total AF in whole chilies were 52 and 38%, compared with values of 54 and 49% in ground chilies respectively. AF contamination was found to be higher in summer chili samples and hence winter chilies may provide a better quality product with respect to AF contamination. The ability to undertake this analysis in Pakistan will enhance greatly the ability to improve chili production in that country, as described herein.     

Deoxynivalenol (DON) in hartweizen

Deoxynivalenol (DON) in Durum Wheat High contamination levels of deoxynivalenol (DON) were found during an investigation of noodles in 2001 and initiated an additional survey of Durum wheat used as raw material for these products. Analyses of 53 samples of Durum wheat by HPLC revealed 89% positives and a median concentration of 790 ug/kg, which clearly exceeded the EU action level of 500 μg/kg. Based on these findings, producers increased quality controls on Durum wheat. In 2002 a total of 60 samples were analysed and 85% positives found. The median, however, lay at 215 μg/kg, below a probable maximum level of 500 μg/kg and demonstrated the positive efforts of food producers to decrease the contamination levels of DON in pasta products.     

Aflatoxin B<Subscript>1</Subscript> contamination in feed from Puglia and Basilicata regions (Italy): 5 years monitoring data

During a 5-year period from 2010 to 2014, n = 919 samples of feed and raw materials were analyzed for aflatoxin B₁ (AFB₁) contamination using accredited ELISA screening methods. Only 0.76 % of these samples were non-compliant with maximum levels set by the European Union Regulation 32/2002. Non-compliant samples were mainly from the province of Bari (n = 3 samples, mean AFB₁ value 7.03 μg/kg), although the highest AFB₁ levels were found in two samples from the provinces of Foggia and Brindisi, at 32.6 ± 3.6 μg/kg and 31.0 ± 4.0 μg/kg, respectively. Mean AFB₁ levels in samples contaminated but compliant with the limits ranged from 1.4 to 2.2 μg/kg. Considering the great importance of climate conditions in mycotoxins production, during crops production and during the critical phases of materials storage and/or transport, to better understand the variability in contamination levels, the analytical results were reviewed in term of temperature and relative environmental humidity in the sampling areas. Correlations between aflatoxin B₁ levels in feed and these climate factors might explain seasonal and annual variations in contamination levels. The data from the present study provide useful suggestions for the organization of targeted monitoring plans and the protection of consumers, as well as for improvement in the quality standards of zootechnological activities and feed industry.