A quantitative detection tool was developed to enable the monitoring of fumonisin-producing fungi in food and feed commodities. To this end, a quantitative PCR (TaqMan) was developed that targets a conserved region in the polyketide synthase gene fum1, which is involved in the biosynthesis of fumonisin. Hence, this method specifically detected isolates from the fumonisin-producing species Fusarium verticillioides, F. proliferatum, F. nygamai and F. globosum whereas isolates of the fumonisin non-producing species F. equiseti, F. graminearum, F. oxysporum, F. semitectum and F. subglutinans that commonly occur on maize were not detected. Moreover, a few fumonisin non-producing F. verticillioides isolates did not generate any fluorescent signals and were therefore not detected. The correlation between quantitative PCR and mycotoxin content was determined using field samples collected at homestead farms in South Africa. Among 40 samples from the Eastern Cape collected in 2005 a good correlation (R2=0.8303) was found between pg fungal DNA and fumonisin content. A similar correlation (R2=0.8658) was found among 126 samples collected from four provinces in South Africa in 2007. These observations indicate that samples containing ≥ 40 pg fungal DNA/mg sample are suspected of also exceeding the 1 mg/kg total fumonisin level and therefore do not comply with the European Commission limit for fumonisins B1+B2 for maize intended for direct human consumption that applies from 1 October 2007. Combined with the very high maize intake, our results indicate that fumonisin levels in maize from South African homesteads regularly exceed the tolerable daily intake for fumonisins.

Fusarium spp. produce fumonisins -mycotoxins that are of importance to maize production in South Africa. Fumonisins have been associated with human oesophageal cancer and cause various diseases in animals that are of concern to the animal feed industry. Maize samples, collected from subsistence farm fields in the Eastern Cape, KwaZulu-Natal, Limpopo and Mpumalanga provinces of South Africa during the 2006 and 2007 growing seasons, were analysed for Fusarium spp. and contamination with fumonisins. Fusarium verticillioides was the most common Fusarium species in maize followed by F. subglutinans and F. proliferatum. Levels of contamination with fumonisins ranged from 0 μg/g to 21.8 μg/g, depending on the region where samples were collected. Levels of fumonisins were highest in northern KwaZulu-Natal (Zululand) where 52% and 17% of samples collected in 2006 and 2007, respectively, exceeded 2 μg/g. Regression analyses showed a positive correlation between fumonisin-producing Fusarium spp. determined by real-time polymerase chain reaction and concentration of fumonisins (r = 0.93). Many samples from Zululand, and some from Mokopane (Limpopo) and Lusikisiki (Eastern Cape), contained fumonisins at levels well above the maximum levels of 2 μg/g set by the Food and Drug Administration (USA) and therefore also the limit of 1 μg/g set by the European Union for food intended for direct human consumption. Regulations governing contamination of grain with fumonisins are not yet implemented in South Africa. The high incidence of fumonisins in subsistence farming systems indicates the need for awareness programmes and further research.

Using micro high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) a simple and fast method for the quantitative determination of 26 mycotoxins was developed. Sample preparation consists of a single extraction step and a dilute-and-shoot approach without further cleanup. With a total run time of 9 min and solvent consumption below 0.3 mL per chromatographic run, the presented method is cost-effective. All toxins regulated by the European Commission with maximum or guidance levels in grain products (fumonisins B1 and B2 (FB1 and FB2)); deoxynivalenol (DON); aflatoxins B1, G1, B2, and G2 (AFB1, AFG1, AFB2, and AFG2); ochratoxin A (OTA); T-2 and HT-2 toxins; and zearalenone (ZEN) can be quantified with this method. Furthermore, the enniatins B, B1, A, and A1 (EnB, EnB1, EnA, and EnA1); beauvericin (BEA); 3-acetyl-deoxynivalenol (3-AcDON); fusarin C (FusC); sterigmatocystin (STC); gliotoxin (GT); and the Alternaria toxins alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT), tentoxin (TEN), and altertoxin I (ATX I) can also be quantified. For all regulated compounds, recoveries ranged between 76 and 120%. For all other toxins, the recovery was at least 51%. The method was applied for the analysis of 42 maize samples from field trials in South Africa.

Mycotoxigenic fungi are common pathogens of maize and groundnuts; they produce mycotoxins which reduce the yield and quality of these grain crops. Numerous agricultural practices including crop rotation and storage methods have been shown to impact mycotoxin accumulation. Therefore, the farming and storage practices in maize and groundnut subsistence farming systems in Pongola, Vryheid, Jozini, Manguzi and Mbazwana Districts of northern KwaZulu-Natal (South Africa) were surveyed to determine their potential role in promoting or mitigating mycotoxin contamination. A questionnaire about agricultural farming practices and storage facilities was presented to 65 subsistence maize and/or groundnut farmers. At least 90% of the farmers surveyed were not aware of mycotoxins and their consequences to animal and human health. The majority of the farmers did not practise crop rotation. However, they practised intercropping and sorted damaged and mouldy grain (maize and groundnuts) before storage. The damaged or mouldy grain was largely used as animal feed, thereby exposing animals to an increased risk of mycotoxicoses. Metal tanks and inqolobane (a type of wooden structure) were identified as the most common storage structures. Harvested homegrown maize was mostly used for the farmers’ own consumption but also sometimes sold to the local community. The implementation of mycotoxin awareness campaigns is necessary, particularly in these districts. The storage facilities used by the subsistence farmers allowed increased moisture and insect invasion. The need for the surveillance of mycotoxins in subsistence-farmed food crops is vital.

No abstract