Mycological and Multiple Mycotoxin Surveillance of Sorghum and Pearl Millet Produced by Smallholder Farmers in Namibia.

Mycological (mycotoxigenic Fusarium and aflatoxigenic Aspergillus spp.) and multiple mycotoxins [aflatoxin B1 (AFB1), fumonisin B (FB), deoxynivalenol and zearalenone] surveillance was conducted on raw whole grain sorghum (Sorghum bicolor) and pearl millet (Pennisetum glaucum) produced on smallholder farms, and processed products sold at open markets in northern Namibia. Fungal contamination was determined with morphological methods as well as with quantitative Real-Time PCR (qPCR). The concentrations of multiple mycotoxins in samples were determined with liquid chromatography tandem mass spectrometry. The incidence of mycotoxigenic Fusarium spp., Aspergillus flavus and A. parasiticus, as well as the concentrations of AFB1 and FB were significantly (P < 0.001) higher in the malts as compared to the raw whole grains, with Aspergillus spp. and AFB1 exhibiting the highest contamination (P < 0.001). None of the analysed mycotoxins were detected in the raw whole grains. Aflatoxin B1 above the regulatory maximum level set by the European Commission was detected in sorghum (2 of 10 samples; 20%; 3-11 µg/kg) and pearl millet (6 of 11 samples; 55%; 4-14 µg/kg) malts. Low levels of FB1 (6 of 10 samples; 60%; 15-245 µg/kg) were detected in sorghum malts and no FB was detected in pearl millet malts. Contamination possibly occurred postharvest, during storage, and/or transportation and processing. By critically monitoring the complete production process, the sources of contamination and critical control points could be identified and managed. Mycotoxin awareness and sustainable education will contribute to reducing mycotoxin contamination. This could ultimately contribute to food safety and security in northern Namibia where communities are exposed to carcinogenic mycotoxins in their staple diet.

Rural Subsistence Maize Farming in South Africa: Risk Assessment and Intervention models for Reduction of Exposure to Fumonisin Mycotoxins.

Maize is a staple crop in rural subsistence regions of southern Africa, is mainly produced for direct household consumption and is often contaminated with high levels of mycotoxins. Chronic exposure to mycotoxins is a risk factor for human diseases as it is implicated in the development of cancer, neural tube defects as well as stunting in children. Although authorities may set maximum levels, these regulations are not effective in subsistence farming communities. As maize is consumed in large quantities, exposure to mycotoxins will surpass safe levels even where the contamination levels are below the regulated maximum levels. It is clear that the lowering of exposure in these communities requires an integrated approach. Detailed understanding of agricultural practices, mycotoxin occurrence, climate change/weather patterns, human exposure and risk are warranted to guide adequate intervention programmes. Risk communication and creating awareness in affected communities are also critical. A range of biologically based products for control of mycotoxigenic fungi and mycotoxins in maize have been developed and commercialised. Application of these methods is limited due to a lack of infrastructure and resources. Other challenges regarding integration and sustainability of technological and community-based mycotoxin reduction strategies include (i) food security, and (ii) the traditional use of mouldy maize.

Detoxification of the Fumonisin Mycotoxins in Maize: An Enzymatic Approach.

Enzymatic detoxification has become a promising approach for control of mycotoxins postharvest in grains through modification of chemical structures determining their toxicity. In the present study fumonisin esterase FumD (EC 3.1.1.87) (FUMzyme®; BIOMIN, Tulln, Austria), hydrolysing fumonisin (FB) mycotoxins by de-esterification, was utilised to develop an enzymatic reduction method in a maize kernel enzyme incubation mixture. Efficacy of the FumD FB reduction method in "low" and "high" FB contaminated home-grown maize was compared by monitoring FB1 hydrolysis to the hydrolysed FB1 (HFB1) product utilising a validated LC-MS/MS analytical method. The method was further evaluated in terms of enzyme activity and treatment duration by assessing enzyme kinetic parameters and the relative distribution of HFB1 between maize kernels and the residual aqueous environment. FumD treatments resulted in significant reduction (≥80%) in "low" (≥1000 U/L, p < 0.05) and "high" (100 U/L, p < 0.05; ≥1000 U/L, p < 0.0001) FB contaminated maize after 1 h respectively, with an approximate 1:1 µmol conversion ratio of FB1 into the formation of HFB1. Enzyme kinetic parameters indicated that, depending on the activity of FumD utilised, a significantly (p < 0.05) higher FB1 conversion rate was noticed in "high" FB contaminated maize. The FumD FB reduction method in maize could find application in commercial maize-based practices as well as in communities utilising home-grown maize as a main dietary staple and known to be exposed above the tolerable daily intake levels.

Marasas et al. 1984 "Toxigenic Fusarium Species: Identity and Mycotoxicology" revisited.

This study was conducted to determine the species identity and mycotoxin potential of 158 Fusarium strains originally archived in the South African Medical Research Council's Mycotoxigenic Fungal Collection (MRC) that were reported to comprise 17 morphologically distinct species in the classic 1984 compilation by Marasas et al., Toxigenic Fusarium Species: Identity and Mycotoxicology. Maximum likelihood and maximum parsimony molecular phylogenetic analyses of single and multilocus DNA sequence data indicated that the strains represented 46 genealogically exclusive phylogenetically distinct species distributed among eight species complexes. Moreover, the phylogenetic data revealed that 80/158 strains were received under a name that is not accepted today (ex F. moniliforme) or classified under a different species name. In addition, gas chromatography-mass spectrometry (GC-MS) and/or high-performance liquid chromatography-mass spectrometry (HPLC-MS)-based mycotoxin analyses were conducted to determine which toxins the strains could produce in liquid and/or solid cultures. All of the trichothecene toxin-producing fusaria were nested within the F. sambucinum (FSAMSC) or F. incarnatum-equiseti (FIESC) species complexes. Consistent with this finding, GC-MS analyses detected trichothecenes in agmatine-containing broth or rice culture extracts of all 13 FSAMSC and 10/12 FIESC species tested. Species in six and seven of the eight species complexes were able to produce moniliformin and beauvericin, respectively, whereas B-type fumonisins were only detected in extracts of cracked maize kernel cultures of three species in the F. fujikuroi (FFSC) species complex.

Biologically Based Methods for Control of Fumonisin-Producing Fusarium Species and Reduction of the Fumonisins.

Infection by the fumonisin-producing Fusarium spp. and subsequent fumonisin contamination of maize adversely affect international trade and economy with deleterious effects on human and animal health. In developed countries high standards of the major food suppliers and retailers are upheld and regulatory controls deter the importation and local marketing of fumonisin-contaminated food products. In developing countries regulatory measures are either lacking or poorly enforced, due to food insecurity, resulting in an increased mycotoxin exposure. The lack and poor accessibility of effective and environmentally safe control methods have led to an increased interest in practical and biological alternatives to reduce fumonisin intake. These include the application of natural resources, including plants, microbial cultures, genetic material thereof, or clay minerals pre- and post-harvest. Pre-harvest approaches include breeding for resistant maize cultivars, introduction of biocontrol microorganisms, application of phenolic plant extracts, and expression of antifungal proteins and fumonisin degrading enzymes in transgenic maize cultivars. Post-harvest approaches include the removal of fumonisins by natural clay adsorbents and enzymatic degradation of fumonisins through decarboxylation and deamination by recombinant carboxylesterase and aminotransferase enzymes. Although, the knowledge base on biological control methods has expanded, only a limited number of authorized decontamination products and methods are commercially available. As many studies detailed the use of natural compounds in vitro, concepts in reducing fumonisin contamination should be developed further for application in planta and in the field pre-harvest, post-harvest, and during storage and food-processing. In developed countries an integrated approach, involving good agricultural management practices, hazard analysis and critical control point (HACCP) production, and storage management, together with selected biologically based treatments, mild chemical and physical treatments could reduce fumonisin contamination effectively. In rural subsistence farming communities, simple, practical, and culturally acceptable hand-sorting, maize kernel washing, and dehulling intervention methods proved to be effective as a last line of defense for reducing fumonisin exposure. Biologically based methods for control of fumonisin-producing Fusarium spp. and decontamination of the fumonisins could have potential commercial application, while simple and practical intervention strategies could also impact positively on food safety and security, especially in rural populations reliant on maize as a dietary staple.