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Mycotoxins are toxins produced by fungi that contaminate many key food crops as they grow in the field and during storage. Specific mycotoxins are produced by different fungi. Each type of fungus and mycotoxin have their own optimal temperatures and water activities for growth and production. The legislative limits for various mycotoxins in foodstuffs to protect human health vary between countries but all commodities have their levels evaluated based on the concentrations from one aggregated grain sample. This approach assumes that the variation in toxin levels is uniform and random without spatial trends. This study investigates the spatial distribution of four mycotoxins (aflatoxin, deoxynivalenol, fumonisin and zearalenone) in bins of clean and dirty corn when stored in an environmental cabinet for two months under different temperature and humidity conditions. The bins of clean and dirty corn each had 12 CO2/humidity/temperature sensors installed in three layers, and samples were extracted for mycotoxin analysis from locations close to each sensor following storage. Using Mann–Whitney U and Kruskal–Wallis H statistical tests, significant differences were found between mycotoxin levels attributable to the different environmental conditions and spatial locations of samples. Variations in aflatoxin and zearalenone concentrations were most pronounced for the range of temperature and humidity conditions chosen. By understanding the patterns of spatial variability in mycotoxin concentrations and identifying zones at high risk of contamination, as well as what conditions are favorable, targeted interventions could be implemented to reduce food waste. This work also has implications for how levels of mycotoxins in foodstuffs are sampled and measured. [ABSTRACT FROM AUTHOR]

Biological control is known to be effective in reducing aflatoxin contamination of corn, and some transgenic corn hybrids incur reduced damage from corn earworm ( Helicoverpa zea) than comparable non- Bt hybrids. We conducted seven field trials over two years to test the hypothesis that transgenic insect protection and biological control could be integrated to prevent mycotoxin contamination of corn. Corn hybrid N78N3111, expressing the highest degree of transgenic insect protection, was nearly 100 percent free from corn earworm damage and generally had less than half as much contamination from fumonisin compared to N78NGT, a near isogenic corn hybrid without insect protection. This insect protection, however, did not significantly prevent aflatoxin contamination. Soil application of non-aflatoxigenic biocontrol strains of Aspergillus flavus significantly reduced aflatoxin concentrations in corn. Biocontrol strain 21882 of A. flavus was especially effective and reduced aflatoxin contamination by about 90 percent over the seven field trials. There was no significant interaction between the insect protection and biocontrol treatments. Although no synergies were detected, the reduction of mycotoxins by both strategies supports application of both strategies in tandem. Economic factors external to the cost of the technologies will be a major determinant if the mycotoxin mitigation attained by use of these technologies will have a positive economic benefit. [ABSTRACT FROM AUTHOR]

Corn infected with Ustilago maydis, causal agent of common smut disease, produces galls that are used as food in certain cultures, but may be contaminated with mycotoxins. The objective of this study was to determine mycotoxin levels in common smut galls (CSGs) collected from the field at corn ear reproductive stages R1 through R5 and in commercial CSGs products. The study was conducted in 2012 and 2013. A simple extraction method for five mycotoxins was devised and the results showed the presence of these compounds in CSGs in corn during ear development at various physiological stages. Fumonisin was the major mycotoxin in CSG samples in both 2012 (63%, ≤150.7 μg g −1 ) and 2013 (46.9%, ≤20.9 μg g −1 ); followed by aflatoxin (2012: 2%, ≤14.7 ng g −1 ; 2013: 30.6%, ≤10.8 ng g −1 ) and zearalenone (2012: ≤41.70 ng g −1 ; 2013: ≤12.40 ng g −1 ). Deoxynivalenol (DON) was only detected in 2012 (≤1.6 μg g −1 ), and cyclopiazonic acid was only detected in 2013 (≤3.18 μg g −1 ). Commercial canned and fresh CSG samples also contained detectable amounts of mycotoxins including aflatoxin, fumonisin, CPA, and DON. Aspergillus flavus was isolated from selected 2013 CSG field samples at R2 or older (0–1.6 × 10 6 cfu/g), whereas Fusarium spp were isolated at R1 or older (0–7.5 × 10 7 cfu/g). These results indicate that CSGs can be infected with mycotoxigenic fungi and contaminated with mycotoxins. The incidence of mycotoxins in commercially available CSG products was highly variable and warrants further study. [ABSTRACT FROM AUTHOR]

Drought stress in the field has been shown to exacerbate aflatoxin contamination of maize and peanut. Drought and heat stress also produce reactive oxygen species (ROS) in plant tissues. Given the potential correlation between ROS and exacerbated aflatoxin production under drought and heat stress, the objectives of this study were to examine the effects of hydrogen peroxide (H2O2)-induced oxidative stress on the growth of different toxigenic (+) and atoxigenic (-) isolates of Aspergillus flavus and to test whether aflatoxin production affects the H2O2 concentrations that the isolates could survive. Ten isolates were tested: NRRL3357 (+), A9 (+), AF13 (+), Tox4 (+), A1 (-), K49 (-), K54A (-), AF36 (-), and Aflaguard (-); and one A. parasiticus isolate, NRRL2999 (+). These isolates were cultured under a H2O2 gradient ranging from 0 to 50 mM in two different media, aflatoxin-conducive yeast extract-sucrose (YES) and non-conducive yeast extract-peptone (YEP). Fungal growth was inhibited at a high H2O2 concentration, but specific isolates grew well at different H2O2 concentrations. Generally the toxigenic isolates tolerated higher concentrations than did atoxigenic isolates. Increasing H2O2 concentrations in the media resulted in elevated aflatoxin production in toxigenic isolates. In YEP media, the higher concentration of peptone (15%) partially inactivated the H2O2 in the media. In the 1% peptone media, YEP did not affect the H2O2 concentrations that the isolates could survive in comparison with YES media, without aflatoxin production. It is interesting to note that the commercial biocontrol isolates, AF36 (-), and Aflaguard (-), survived at higher levels of stress than other atoxigenic isolates, suggesting that this testing method could potentially be of use in the selection of biocontrol isolates. Further studies will be needed to investigate the mechanisms behind the variability among isolates with regard to their degree of oxidative stress tolerance and the role of aflatoxin production. [ABSTRACT FROM AUTHOR]

Biological control of the fungus Aspergillus flavus has been shown to be effective in reducing aflatoxin contamination in corn. This study compared field application of a bioplastic-based formulation for delivering atoxigenic A. flavus isolates in Northern Italy and the Mississippi Delta. Due to an extremely hot and dry summer at the Italy site in 2012, aflatoxin contamination was approximately seven times higher than in 2011. In 2011, and 2012, application of bioplastic granules inoculated with the atoxigenic isolate A. flavus NRRL 30797 at 15 and 30 kg ha −1 resulted in a reduction of aflatoxin contamination by 67.2 ± 4.1% and 94.8 ± 5.3%, respectively. The higher application rate was also effective when soil abundance of A. flavus was artificially increased by applying contaminated corn residues. At the Mississippi site, summer 2012 was also hot and dry, with high levels of aflatoxin contamination. In fields planted with non-Bt or Bt hybrids, application of biocontrol granules inoculated with A. flavus NRRL 30797 or NRRL 21882 at 30 kg ha −1 reduced aflatoxin contamination to up to 89.6%. Field experiments on two continents showed that bioplastic-based A. flavus formulations markedly reduced aflatoxin contamination under different agro-environmental conditions and infestation intensities. [ABSTRACT FROM AUTHOR]

Abstract: Contamination of corn, cotton, peanuts and tree nuts by aflatoxins is a severe economic burden for growers. A current biocontrol strategy is to use non-aflatoxigenic Aspergillus flavus strains to competitively exclude field toxigenic Aspergillus species. A. flavus K49 does not produce aflatoxins and cyclopiazonic acid (CPA) and is currently being tested in corn-growing fields in Mississippi. We found that its lack of production of aflatoxins and CPA resulted from single nucleotide mutations in the polyketide synthase gene and hybrid polyketide-nonribosomal peptide synthase gene, respectively. Furthermore, based on single nucleotide polymorphisms of the aflatoxin biosynthesis omtA gene and the CPA biosynthesis dmaT gene, we conclude that K49, AF36 and previously characterized TX9-8 form a biocontrol group. These isolates appear to be derived from recombinants of typical large and small sclerotial morphotype strains. This finding provides an easy way to select future biocontrol strains from the reservoir of non-aflatoxigenic populations in agricultural fields. [Copyright &y& Elsevier]

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Abstract: A severe infestation by aflatoxin-producing fungi diminished food quality of southern United States corn (maize) in 1998. Corn hybrids (65) naturally infected with Fusarium spp. and Aspergillus spp. were evaluated from 1998 to 2001 for resistance to mycotoxin contamination. Kernel corn samples were assayed at harvest for aflatoxins and fumonisins. In 1998, samples from all hybrids exceeded 20ppb aflatoxin (mean levels: 21–699ppb) and 2ppm fumonisins (mean levels: 23–79ppm), the maximum levels permitted by United States Food and Drug Administration guidelines. Samples from hybrids planted in the same and other locations in Arkansas in 1999 and 2001 were shown by similar methods to contain aflatoxin levels ranging from not detected to 255.3ppb and fumonisin levels from 0.3 to 83.6ppm. The fumonisin levels in 2001 were very high in all hybrids, ranging from 8 to 83.6ppm while aflatoxin levels were low ranging from <5 in most hybrids to 131ppb. The presence of aflatoxin B1 and B2 in samples was confirmed by thin layer chromatography and liquid chromatography/mass spectrometry and fumonisins B1, , , and by liquid chromatography/mass spectrometry. During the period studied, a positive correlation was observed between aflatoxin and fumonisin levels, indicating that natural infection with Fusarium spp. did not appear to protect against aflatoxin production. [Copyright &y& Elsevier]

Aflatoxins present important food safely problems in both developed and developing countries. Contamination is monitored in developed countries using enzyme-linked immunusorbent assay(ELISA)- and high-performance liquid chromatography(HPLC)-based assays, both of which may be too expensive for routine use in many developing countries. There is a need for inexpensive alternative approaches to detect aflatoxins in lots of foods and feeds. Reviewed here are culture-based methods that determine if a sample is contaminated with aflatoxigenic fungi. These approaches include 1) blue fluorescence of aflatoxin B1, particularly when enhanced by includingβ-cyclodextrin in the culture medium, 2) yellow pigment production, and 3) color change on exposure to ammonium hydroxide vapor. The presence of aflatoxin B1 can be detected by its blue fluorescence, which is enhanced when the toxin complexes with the hydrophobic pocket ofβ-cyclodextrin. The yellow pigment and ammonium hydroxide vapor tests are based on the production of yellow anthraquinone biosynthetic intermediates in the aflatoxin pathway. These compounds act as pH indicator dyes, which are more visible when they have turned red at alkaline pH. Because these tests are based on two different mechanisms, it has been possible to combine them into a single test. In a study of 517 A. flavus isolates from the Mississippi Delta, the combined assay reduced false positives for aflatoxigenicity to 0%, and false negatives to 7%. The increased predictive power of the combined cultural assay may enable its use for inexpensively identifying potential aflatoxin contamination in feeds and foods. [ABSTRACT FROM AUTHOR]

Abstract: Studies on biological control of aflatoxin production in crops by pre-infection with non-toxigenic Aspergillus flavus strains have created a need for improved methods to screen isolates for aflatoxigenicity. We have evaluated two empirical aflatoxigenicity tests: (i) yellow pigment production, and (ii) the appearance of a plum-red color in colonies exposed to ammonium hydroxide vapor. Yellow pigments from aflatoxigenic A. flavus were shown to function as pH indicator dyes. Seven pigments representing most of the pigmentation in extracts have been isolated using color changes when chromatography spots were exposed to ammonium hydroxide vapor to guide fractionation. Their structures have been shown to be norsolorinic acid, averantin, averufin, versicolorin C, versicolorin A, versicolorin A hemiacetal and nidurufin, all of which are known anthraquinone pigments on, or associated with, the aflatoxin biosynthetic pathway in Aspergillus spp. Thus, the basis of both empirical tests for aflatoxigenicity is detecting production of excess aflatoxin biosynthetic intermediates. [Copyright &y& Elsevier]