Mycoflora and mycotoxins natural occurrence in corn from entre rios province, Argentina

Corn samples were collected in 1999 from three departments of Entre Réos province, Argentina, and were surveyed for mould contamination and natural occurrence ofFusarium mycotoxins, ochratoxin A and aflatoxins.Fusarium verticillioides was the most prevalent fungal species recorded at all departments. Zearalenone, deoxynivalenol and ochratoxin A were not found in any samples. Only one of the 52 corn samples analysed was contaminated with aflatoxin B₁ (17 μg/kg). Fumonisin B₁ was found in 58 % of samples (range of positive samples: 47– 3,347 μg/kg), fumonisin B₂ in 33.0 % (range of positive samples: 23–537 μg/kg) and fumonisin B₃ in 25.0 % (range of positive samples: 24–287 μg/kg) of them. This is the first report on the natural occurrence of mycotoxins in corn from Entre Ríos province, Argentina. Levels of fumonisins were lower than detected in other Argentinian provinces.     

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.     

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)     

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.     

Mycotoxicological control on raw material and tablets of cascara sagrada (Rhamnus purshiana)

Among phytotherapic medicines, tablets of cascara sagrada (Rhamnus purshiana) dried bark, usually used as laxative, are commercially widespread in our market. Taking into account natural origin and/or inappropriate procedures that may allow the occurrence of toxinogenic Aspergillus flavus group, a study on susceptibility to aflatoxin contamination and natural aflatoxin incidence was performed by TLC and HPLC methods. This survey allows one to conclude that bark of Cascara Sagrada is a good substrate for the growth of A. parasiticus NRRL 2999 and for aflatoxins production. Natural anatoxins presence was detected on 2 from 9 raw material samples. One of them (irradiated sample) had only aflatoxin B1 (10 μg/kg) and the other (pasteurized) was positive for aflatoxin B1 (19 μg/kg); G1 (6 μg/kg) and B2 (1.46 μg/kg). Only one from 10 lots of tablets analyzed was positive for aflatoxin B1 (5.42 μg/kg) and B2 (0.32 μg/kg).Therefore, adequate quality control including an aflatoxins assay must be performed to guarantee the harmlessness of natural drugs.     

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.     

Incidence and levels of fumonisin contamination in Colombian corn and corn products

A survey was conducted to determine the levels of fumonisins B1 and B2 in corn and corn-based products available in Colombia for human and animal consumption. A total of 120 samples were analyzed by acetonitrile-water extraction, cleanup with a strong-anion-exchange column, and liquid chromatography with o-phthaldialdehyde-2-mercaptoethanol derivatization and fluorescence detection. The samples of corn and corn-based products for animal intake were taken at different feed manufacturing plants, whereas the samples used for human foods where purchased from local retail stores. The number of positive samples for fumonisin B1 was 20.0% higher in corn and corn-based products for animal intake (75.0%) than in corn and corn-based products for human consumption (55.0%). The levels of fumonisin B1 were also higher in corn and corn-based products for animal intake (mean = 694 μg/kg; range = 32–2964 μg/kg), than in corn and corn-based products for human intake (mean = 218 μg/kg; range = 24–2170 μg/ kg). The incidence and levels of fumonisin B2 were lower than those for fumonisin B1. Corn and corn-based products for animal consumption had an incidence of fumonisin B2 of 58.3%, with a mean value of 283 μg/kg, and a range of 44–987 μg/kg. The incidence of fumonisin B2 in corn-based products for human intake was 35.0%, with a mean value of 118 μg/kg and a range of 21–833 μg/kg. The highest incidence and levels of fumonisins were found in samples of hominy feed, with concentrations ranging from 86 to 2964 μg/kg fumonisin B1 and 57 to 987 μg/kg fumonisin B2.     

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.     

Factors affecting the selenium intake of people in transbaikalian Russia

The selenium concentration in foods grown and consumed and in plasma, red blood cells, and toenails of people living in the district of Chita in the transbaikalian part of Russia were studied in August 1991. Preliminary results from the area have suggested low selenium intakes and the possible occurrence of cardiomyopathy (Keshan disease) in the population. A low selenium concentration in foods grown locally was found: mean selenium concentration in wheat grains was 1, 5, and 28 μg/kg, respectively, in three villages studied, that of oats was beween 3–6 μg/kg, and of cow's milk 10–27 μg/kg dry matter. The selenium concentration of bread was considerably higher, between 87–337 μg/kg dry wt, presumably because wheat imported from the US had been used for baking. Occasional samples of pork, beef, and mutton contained between 32–318 μg selenium/kg dry wt. Low selenium concentrations were observed in samples of soil and river water. The mean plasma selenium concentration of 52 persons was 1.02 μmol/L, including 33 children and 19 adult subjects. The selenium concentrations in red blood cells and toenails were 1.95 μmol/L and 0.61 mg/kg, respectively. No symptoms of heart disease caused by selenium deficiency were observed. It is concluded that the selenium status of people was fairly good thanks to the contribution to dietary intake of imported wheat with a high selenium content. As the selenium concentration was very low in foods grown in the area, the selenium intake of the population will be reduced to a very low level if only locally produced foods are consumed.     

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     

Potential pathogenicity of Cladosporium tennuisimum,Phaeoisaria clematidis and Ramichloridium subulatum in a mouse model

In Sri Lanka, rice is the main staple which is mostly processed into parboiled rice. The levels of aflatoxin B₁ (AFB₁) and aflatoxin G₁ (AFG₁) in parboiled and raw milled rice collected from major rice producing areas and rice consuming townships were estimated. In almost all the samples of parboiled rice examined, the AFB₁ and AFG₁ contents were significantly higher than in raw milled rice. The highest AFB₁ content was 185 µg/kg and AFG₁ content 963 g/kg. These samples were collected from a major rice producing/milling district where the mean relative humidity is 78% and mean annual temperature 27 °C which is the highest amongst the rice growing areas in Sri Lanka. Raw rice was either free of aflatoxins or when toxins were detected, they occurred in less than 10% of the samples. The frequency of occurrence of surface fungal flora (Aspergillus/Penicillium) and aflatoxin content in market samples was closely related. Brownish or greenish moldy rice samples with fermented odour contained over 1000 g/kg of AFB₁.     

A survey of aflatoxin B<Subscript>1</Subscript> and total aflatoxin contamination in baby food,peanut and corn products sold at retail in Indonesia analysed by ELISA and HPLC

Aflatoxin contamination has been well known as a world-wide health-threatening problem in tropical countries including Indonesia. This research was undertaken to determine the degree of aflatoxin contamination in different Indonesian foodstuffs. A preliminary survey was carried out to evaluate the level of total aflatoxin (AfT) and aflatoxin B₁ (AfB₁) contamination of baby foods, peanut products, and corn products, which were purchased from traditional markets and supermarkets in Indonesia during the year 2001-2002. Eighty two peanut products, 12 baby foods products, and 11 corn products from different brands were analysed for AfT and AfB₁ using the Enzyme-Linked Immunosorbent Assay (ELISA) method. The results indicate that, of the brands analysed, 35% of the peanut products were contaminated with aflatoxins at various levels (range 5 to 870 μg/kg). Peanut-chilli sauces had the highest percentage of AfT contamination 9/12 (75%), which was followed by traditional snacks 5/11 (45%), peanut butter 4/11 (40%), flour egg coated peanut 6/16 (37%), and peanut cake 3/10 (30%). Fried peanuts and roasted peanut were found to contain aflatoxin at relatively lower percentages of 9% and 8%, respectively. From the 12 analysed baby food samples, on the other hand, no sample was found to be contaminated with aflatoxins. Two of 11 samples (18%) of corn based products were contaminated with AfT, ranging between 5.8 and 12.4 μg/kg. Additionally, 30 selected samples in different concentration ranges were further analysed to verify the correlation between ELISA and HPLC techniques and results were compared.     

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%).     

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.     

Aflatoxin content and number of fungi in poultry feedstuffs from Indonesia

The content of aflatoxin and associated fungi was determined in 56 samples, including 34 of corn, 10 of soybean meal, nine of rice bran and three of broken rice, collected from different poultry farms and poultry feedmills situated around Jakarta-Bogor, Indonesia.
Ninety-one per cent of the corn samples contained aflatoxins and the total concentration ranged from 22 to 6171 μ g/kg. With rice bran, 100% of the samples were positive for aflatoxin B₁, ranging from 36 to 71 μ g/kg. No aflatoxin was detected in samples of soybean meal or broken rice. All the samples were contaminated by several fungi (8 times 10³–5 times 10⁶cfu/g) and further identification was limited to Aspergillus flavus and A. parasiticus. The dominant species was A. flavus (2 times 10³–4 times 10⁶cfu/g in corn samples, 1·0 times 10³–1·0 times 10⁵cfu/g in soybean meal, 2 times 10⁴–4·4 times 10⁵cfu/g in rice bran and 2 times 10⁴–6 times 10⁴cfu/g in broken rice). Some of the corn samples also contained A. parasiticus (2 times 10³–9·5 times 10⁴cfu/g).     

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.     

Aflatoxin B<Subscript>1</Subscript> in chilies from the Punjab region,Pakistan

The occurrence of aflatoxin B1 (AFB1) in chilies from Pakistan was determined by using HPLC in work undertaken in Pakistan. Whole (n = 22) and powdered (n = 22) chilies were analyzed. Sixteen (73.0%) and 19 (86.4%) samples of whole and ground chilies, respectively, were contaminated. The mean concentration in powdered chilies (32.20 μg/kg) was higher statistically than in whole chilies (24.69 μg/kg). Concentrations ranged from 0.00 to 89.56 μg/kg for powdered chilies, compared with 0.00–96.3 μg/kg for whole chilies. The limits of detection and quantification were 0.05 μg/kg and 0.53 μg/kg, respectively. The concentrations were high in general and greater than the statutory limit set by the European Union. There is considerable scope for improvements in chili production in Pakistan.     

Development of a method for the determination of citrinin in barley,rye and wheat by solid phase extraction on aminopropyl columns and HPLC-FLD

A method for the determination of the mycotoxin citrinin (CT) in rye, wheat and barley is described. The proposed method is based on ethyl acetate extraction, solid phase clean-up (SPE) on aminopropyl columns and reversed phase high performance liquid chromatography with fluorescence detection (RP-HPLC-FLD). The limits of detection and quantification of CT amounted to 0.6–0.9 μg/kg and 1.7– 3.3 μg/kg with mean recovery rates in the range of 77–92% (RSD 4.8–5.5%). This method can also be used for the determination of CT in red-fermented rice. Presented at the 29^th Mykotoxin-Workshop, Fellbach, Germany, May 14–16, 2007     

Natural Occurrence of Mycotoxins in Staple Cereals from Ethiopia

The occurrence of mycotoxins in barley, sorghum, teff (Eragrostis tef) and wheat from Ethiopia has been studied. Samples were analyzed for aflatoxin B₁ (AFB1), ochratoxin A (OTA), deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN) using high performance liquid chromatography (HPLC) and for fumonisins (FUM) using enzyme linked immunosorbent assay (ELISA). AFB1 and OTA were detected in samples of all the four crops. AFB1 was detected in 8.8% of the 352 samples analyzed at concentrations ranging from trace to 26 μg kg⁻¹. OTA occurred in 24.3% of 321 samples at a mean concentration of 54.1 μg kg⁻¹ and a maximum of 2106 μg kg⁻¹. DON occurred in barley, sorghum and wheat at 40–2340 μg kg⁻¹ with an overall incidence of 48.8% among the 84 mainly ‘suspect’ samples analyzed; NIV was co-analyzed with DON and was detected at 40 μg kg⁻¹ in a wheat sample and at 50, 380, and 490 μg kg⁻¹ in three sorghum samples. FUM and ZEN occurred only in sorghum samples with low frequencies at concentrations reaching 2117 and 32 μg kg⁻¹, respectively. The analytical results indicate higher mycotoxin contamination in sorghum, which could be related to the widespread storage of sorghum grain in underground pits leading to elevated seed moisture contents. This is the first report on the occurrence of OTA in teff.     

Aflatoxin contamination of pearl millet during field and storage conditions with reference to stage of grain maturation and insect damage

Aflatoxin contamination in five varieties of pearl millet (ICMH-451, ICMP-50I, ICTP-8203, WCC-75 and ICMV-155) was studied from field and storage conditions in three districts of Andhra Pradesh State, India and the inter-relationships between various parameters such as stage of grain maturation in the field and insect pest infestation in storage in relation to aflatoxin production were evaluated. Aflatoxin contamination was more frequent in the seed samples collected from the fields during rainy season than winter season. All major aflatoxins were isolated from one or the other varieties of pearl millet, whereas aflatoxin G₂ was not commonly observed in the seed samples collected during winter. Among all the varieties tested, ICMH-451 was vulnerable to aflatoxin contamination whereas ICMV-155 was the least susceptible variety. The higher amount of aflatoxins was observed in the matured seed samples followed by pre-matured and milky stage. Among all the toxins reported in the field, aflatoxin B₁ was found in higher concentration (185 (μg/kg) followed by B₂ (105 μg/kg). The four major types of aflatoxins with higher levels (35, 40, 140, 190 μg/kg of G₁, G₂, B₂, B₁ were reported in the rainy season seed samples after six months of storage, whereas aflatoxin G₁ was not observed in any variety of stored seed sample from winter. Statistical analysis revealed that the aflatoxin incidence in relation to different parameters studied was significantly different for each factor. The relationship between aflatoxin contamination and insect damaged-grain clearly indicated that the seed samples with 16-40% of insect damage contained higher amounts of aflatoxins (758 μg/kg). Presented at the 29^th Mykotoxin-Workshop, Fellbach, Germany, May 14–16, 2007