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B-series fumonisins (FBs) are a family of carcinogenic mycotoxins that commonly occur in maize. These mycotoxins cause multiple diseases in animals and are epidemiologically associated with several human diseases in populations for which maize is a dietary staple. FBs are produced by multiple genera of the fungi Aspergillus, Fusarium and Tolypocladium, but the plant pathogen Fusarium verticillioides is considered the primary cause of FB contamination in maize. One F. verticillioides strain, MRC 826, is reported to produce high levels of FBs. However, in the current study, 18 isolates derived from strain MRC 826 exhibited highly variable levels of FB, which negatively correlated (r=-0.333; P

Differential anti-proliferative and pro-apoptotic effects of aqueous extracts of green rooibos (Rg; Aspalathus linearis) and green tea (GT; Camellia sinensis) and an aspalathin-enriched extract of green rooibos (GRE), were investigated in primary rat hepatocytes (PH) and human liver (HepG2) and colon (HT-29) cancer cells. Rooibos flavonoids, aspalathin and luteolin, and the green tea flavanol, epigallocatechin gallate (EGCG), were included to assess their contribution relative to their extract concentrations. GRE was the most effective in reducing cell growth parameters which was associated with a high total polyphenol content and high ferric reducing potential. Differential cell responses were noticed with HepG2 cells more sensitive than PH toward the induction of apoptosis by GRE. Luteolin induced apoptosis in PH and HepG2 cells while aspalathin lacked any effect. EGCG induced apoptosis in HepG2 cells while PH were resistant. HT-29 cells were resistant to apoptosis induction by the tea and pure flavonoids. Differences existed in the individual effects of the major rooibos and GT flavonoids against cell growth parameters compared to their equivalent concentrations in the extract mixtures. Diversity of the flavonoid constituents, physicochemical properties and cellular redox status governing cell survival are likely to explain the differential cell responses.

The formal honeybush tea industry has reached a 20-year milestone on 24 February 2019, giving an opportunity to reflect on the research targeting phenolic composition, quality and bioactivity, since a previous review published in 2008. This research underpins ongoing endeavours to unlock the potential of this indigenous agricultural commodity. The current review illustrates that the bulk of the research followed a clear ‘vision’ to enhance consumer confidence in the product and by exploring value-adding opportunities. Gaps in knowledge were highlighted that could drive future research directions.

The fumonisins are carcinogenic mycotoxins that have been implicated in various adverse human health effects. Recently, the Codex Alimentarius Commission set maximum levels (MLs) for fumonisin mycotoxins at 4000 μg/kg for raw maize and 2000 μg/kg for maize flour and maize meal and these have been incorporated into South African health regulations. The MLs are intended to be health protective such that fumonisin exposures will be below the provisional maximum tolerable daily intake (PMTDI) of fumonisin of 2 μg/kg body weight/day set by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). South Africa has an extensive commercial maize farming industry as well as a large subsistence farming population reliant on home-grown maize. The available data on fumonisin contamination levels indicates that these MLs are readily met by the commercial crop. However, the situation in subsistence farming areas such as in the Eastern Cape Province region is of concern. The daily consumption of large amounts of highly fumonisin-contaminated home-grown maize is of relevance mainly due to rudimentary milling, which is not subject to the reduction in mycotoxin contamination associated with commercial milling. Analysis of historical fumonisin data from the Eastern Cape region indicates that approximately 12.8% of raw maize and 25% of the rural locally produced flour would not meet the respective MLs of 4000 μg/kg ML and 2000 μg/kg. Further analysis shows that given the high maize intake levels of this population, only prohibitively low MLs for raw maize of around 100–300 μg/kg would be health protective for different age groups in this area. Based on this analysis, the lowering of fumonisin exposure in subsistence farmers requires an integrated approach and cannot be achieved purely by regulatory means.

Fumonisin B1 (FB1), a group 2B natural occurring carcinogenic mycotoxin, modulated lipid and fatty acid (FA) constituents of lipid rafts isolated from primary hepatocytes following exposure to a cytotoxic concentration of FB1 (250 μM). The major effects observed in rafts, included a significant (p

Populations consuming aflatoxin (AF) and fumonisin (FN)-contaminated foods may be at increased risk for hepatocellular carcinoma (HCC) and developmental disorders; consequently, development of intervention strategies to reduce AF/FN-induced liver disease and adverse health effects in humans could be very useful. Calcium montmorillonite clay (NovaSil) has been shown to absorb AF in vitro, in multiple animal models, as well as in human studies. In the present study, we aimed to evaluate whether uniform particle size NovaSil (UPSN) possessed an ability to modulate the co-carcinogenic potentials of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) in F344 rats. Sequential treatment of FB1 following AFB1 synergistically induces preneoplastic alterations as well as liver damage, indicating that AFB1 acts as an initiator while FB1 as a promoter in the carcinogenesis model, confirming findings from previous studies. The enterosorbent agent UPSN clay at dose of up to 0.5% in diet was shown to be effective in modulating the toxicity and carcinogenicity of co-exposure to AFB1 and FB1, as demonstrated by significant reduction in number and size of hepatic GST-P+ foci, in alterations indicative of liver toxicity, and in levels of AFB1 and FB1 biomarkers.

Antimutagenesis studies against the tobacco-specific mutagens 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N-oxide, 4 (methyl-nitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), were conducted using hot water aqueous extracts of rooibos ( Aspalathus linearis ), honeybush ( Cyclopia intermedia ), and cancer bush ( Sutherlandia frutescens ). Aqueous extracts of both “fermented” and “unfermented” (green) rooibos and honeybush were included, while extracts of green and black teas ( Camellia sinensis ) served as benchmarks. A polyphenol-enriched methanol extract of unfermented rooibos (RgM) was included to further elucidate the possible role of rooibos polyphenols. Studies were performed in the presence of the metabolic activation against Salmonella typhimurium tester strain TA1535, using the standard plate incorporation and micro-suspension, pre-incubation assays. The mutagenic effects of NNK against the strain TA1535 was best demonstrated using the standard plate incorporation assay, while a higher mutagenicity was demonstrated for NNAL using the micro-suspension, pre-incubation method. Black tea and RgM exhibited the highest protection against NNK-induced mutagenesis followed by the aqueous extracts of rooibos ≥ green tea ≥ honeybush ≥ cancer bush. Black tea, green tea, RgM and unfermented rooibos were the most effective against NNAL-induced mutagenesis, followed by fermented rooibos. The two honeybush extracts exhibited similar, but the weakest protective response. When considering the amount of total polyphenols (TPP) incorporated in the plate incorporation assay, cancer bush exhibited similar protection to that of fermented and unfermented honeybush against NNK mutagenesis. The involvement of specific polyphenol-cytochrome P450 (CYP450) interactions is likely to be involved in the protection against tobacco-related mutagenesis. Polyphenol constituents of rooibos, honeybush and cancer bush could play an important role in the protection against mutagenesis induced by the major tobacco-specific carcinogens.

Interest in Cyclopia spp. (honeybush) as a source material for production of polyphenol-enriched extracts (PEEs) for the food ingredient and nutraceutical markets requires investigation of their safety. PEEs of Cyclopia subternata (PECsub) and Cyclopia genistoides (PECgen) were fed (2.5 g/kg feed) to male Fischer rats for 28 days, while PECsub, having the highest total polyphenol content and antioxidant activity, was also fed for 90 days. Their dietary intake did not significantly (P ≥ 0.05) affect body weight gain or relative liver and kidney weight. PECsub resulted in a significant (P

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&reg, 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 &ldquo, low&rdquo, and &ldquo, high&rdquo, 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 (&ge, 80%) in &ldquo, (&ge, 1000 U/L, p &lt, 0.05) and &ldquo, (100 U/L, p &lt, 0.05, &ge, 0.0001) FB contaminated maize after 1 h respectively, with an approximate 1:1 &micro, mol conversion ratio of FB1 into the formation of HFB1. Enzyme kinetic parameters indicated that, depending on the activity of FumD utilised, a significantly (p &lt, 0.05) higher FB1 conversion rate was noticed in &ldquo, 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.

What are the molecular events that fumonisin-induced porcine pulmonary edema syndrome and equine leucoencephalomalacia have in common? Do these animal diseases relate mechanistically to fumonisin toxicity in laboratory rats? There is considerable data indicating that disruption of sphingolipid metabolism plays an important early role in all of these diseases. In vitro studies have revealed that fumonisins and structurally related Alternaria alternata f. sp. lycopersici-toxin (AAL-toxin) are potent inhibitors of the enzyme sphinganine (sphingosine) N-acyl transferase (ceramide synthase). Soon after cultured cells or animals are exposed to fumonisins there is a dramatic increase in the free sphingoid base, sphinganine, in tissues, serum and/or urine. Also, free sphingosine concentration increases, complex sphingolipid concentration decreases, and sphingoid base degradation products and other lipid products also increase. It is hypothesized that disruption of sphingolipid metabolism is an early molecular event in the onset and progression of cell injury and the diseases associated with consumption of fumonisins. However, the exact mechanisms responsible for the diseases will not be easily revealed since the role of sphingolipids in cellular regulation is very complex and not yet fully understood. While fumonisin B1 is non-genotoxic it is a complete carcinogen in rat liver. Recent studies indicate that fumonisins inhibit hepatocyte proliferation in rat liver. It has been hypothesized that hepatotoxicity and effects on hepatocyte proliferation are critical determinants for fumonisin B1 cancer initiation and promotion. Alternatively, recent studies have found that fumonisin B1 has mitogenic activity in cultured fibroblasts. It is conceivable that the mitogenic, cytostatic and cytotoxic potential of fumonisin may all contribute to the animal diseases including liver cancer in rats.

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.

Fumonisin esterase FumD (EC 3.1.1.87), FUMzyme® (BIOMIN, Austria), effectively detoxifies fumonisin B mycotoxins (FB) by hydrolysis and removal of the tricarballylic acid groups. The current study evaluated FumD detoxification of total FB (FBT) in commercial maize utilising an experimental dry milling plant by introducing the enzyme during the kernel conditioning stage. Total FB and the hydrolysed product of FB1, HFB1, in maize and milling products were determined by LC-MS/MS. During maize conditioning of 4 h 10 min substantial FB1 hydrolysis was achieved between 1 and 4 U FumD/100 g maize. Complete conversion into HFB1 was delayed and only achieved at the highest enzyme concentration (32 U/100 g maize) reaching a 1:1 M conversion ratio. Dry milling of maize containing 3.29 ± 0.20 μmole FBT/kg (2354 ± 140 μg/kg) in the absence of FumD, resulted in a 2.5-fold increase in the FBT concentration in total hominy feed (8.34 ± 0.22 μmol/kg) (5979 ± 158 μg/kg) compared to the levels that prevail in Super (0.52 ± 0.07 μmol/kg) (347 ± 48 μg/kg) and Special (1.70 ± 0.01 μmol/kg) (1213 ± 8 μg/kg) maize meal, and Semolina (1.07 ± 0.14 μmol/kg) (765 ± 100 μg/kg) milling products. Introduction of FumD (40 U/kg) mainly impacted the total hominy feed product (germ + hominy milling fractions), constituting up to 30% of the reconstituted whole maize. A 99% reduction in FBT was obtained in total hominy feed, 48% in Semolina, 7% in Special maize meal, whereas no reduction was recorded in Super maize meal. FB1 reduction rates depend on the contamination level, kernel moisture and the diffusion rate from inner kernel layers to the kernel surface/aqueous interface. Risk modelling in children and adults indicated that FumD-treated whole maize and the resultant Semolina milling product intended for human consumption reduces the risk of exposure to FBT. However, no reduction in the exposure risk was observed when considering the Super and Special maize meal milling products. FB reduction in total hominy feed could open up new applications, such as its dietary incorporation as a source of fibre, minerals and bioactive plant constituents in maize-based food. In addition, the animal feed industry and subsistence maize farming communities using rudimentary milling processes, could also benefit.

In developing countries the enforcement of compliance to detailed mycotoxin regulations ensures protection of the population from adverse health effects of mycotoxin exposure. In low-income or developing countries mycotoxin regulations are either lacking or poorly enforced which create scenarios where mycotoxin exposures occur above levels set by health regulatory bodies. Population groups that are the worst affected include subsistent maize growing farmer communities where mono-cereal crops are cultivated and locally consumed, and mycotoxin contamination are not monitored. Other factors that aggravate the situation include the consumption of highly mycotoxin contaminated unprocessed maize, the lack of knowledge about the adverse effects as well as traditional uses of maize products not intended for human consumption during periods of food insecurity. These scenarios require ingenious ways to reduce mycotoxin exposure in poor rural communities where access to sophisticated mycotoxins reduction techniques is not available or practically viable. Although community-based and culturally acceptable methods have, to some extent, been adapted the efficacy thereof varies due to the lack of sufficient training. Integration of these methods with more sophisticated technological methods is envisaged, and will be based on a better understanding of mycotoxin biosynthesis and fungus-host interactions on a molecular level. In addition, other methods which include the detoxification of mycotoxins utilising degradation enzymes, clay adsorbents, utilisation of non-toxigenic fungal strains and resistant maize cultivars to fungal infections are just a few approaches under scrutiny. The introduction of good agriculture practices and storage techniques and the identification of critical control points during hazard analyses need to be further explored. Introduction of mycotoxin monitoring programs and validated screening procedures to monitor exposure should be a priority in the future, to facilitate community-based and effective intervention programmes of mycotoxin reduction.

Objectives The relationship between polyphenol constituents, antioxidant properties of aqueous and methanol extracts of green tea (Camellia sinensis), the herbal teas, rooibos (Aspalathus linearis) and honeybush (Cyclopia spp.), against skin cell viability was investigated in vitro. Methods The effect of extracts, characterised in terms of polyphenol content and antioxidant properties, on cell viability of premalignant, normal and malignant skin cells was determined. Key findings Phenolic composition, particularly high levels of potent antioxidants, of rooibos and green tea methanol extracts was associated with a strong reduction in cell viability specifically targeting premalignant cells. In contrast, the aqueous extracts of Cyclopia spp. were more effective in reducing cell viability. This correlated with a relatively high flavanol/proanthocyanidin content and ABTS radical cation scavenging capacity. The major green tea flavanol (epigallocatechin gallate) and rooibos dihydrochalcone (aspalathin) exhibited differential effects against cell viability, while the major honeybush xanthone (mangiferin) and flavanone (hesperidin) lacked any effect presumably due to a cytoprotective effect. The underlying mechanisms against skin cell viability are likely to involve mitochondrial dysfunction resulting from polyphenol–iron interactions. Conclusions The polyphenol constituents and antioxidant parameters of herbal tea extracts are useful tools to predict their activity against skin cell survival in vitro and potential chemopreventive effects in vivo.

Differential sensitivity of primary hepatocytes and Chang cells to the cancer promoter fumonisin B1 (FB1)-induced cytotoxic effects were investigated in relation to changes in membrane lipid distribution. In contrast to primary hepatocytes, Chang cells were resistant to FB1-induced cytotoxic effects. This was associated with a high cholesterol (Chol) and sphingomyelin (SM) and low phosphatidylcholine (PC) content, resulting in a significant (P

International audience; Fumonisin B1 (FB1), a congener of fumonisins produced by Fusarium species, is the most abundant and most toxicologically active fumonisin. FB1 causes severe mycotoxicosis in animals, including nephrotoxicity, hepatotoxicity, and disruption of the intestinal barrier. However, mechanisms associated with FB1 toxicity are still unclear. Preliminary studies have highlighted the role of liver X receptors (LXRs) during FB1 exposure. LXRs belong to the nuclear receptor family and control the expression of genes involved in cholesterol and lipid homeostasis. In this context, the toxicity of FB1 was compared in female wild-type (LXR+/+) and LXR, double knockout (LXR-/-) mice in the absence or presence of FB1 (10mg/kg body weight/day) for 28days. Exposure to FB1 supplemented in the mice's drinking water resulted in more pronounced hepatotoxicity in LXR-/- mice compared to LXR+/+ mice, as indicated by hepatic transaminase levels (ALT, AST) and hepatic inflammatory and fibrotic lesions. Next, the effect of FB1 exposure on the liver transcriptome was investigated. FB1 exposure led to a specific transcriptional response in LXR-/- mice that included altered cholesterol and bile acid homeostasis. ELISA showed that these effects were associated with an elevated FB1 concentration in the plasma of LXR-/- mice, suggesting that LXRs participate in intestinal absorption and/or clearance of the toxin. In summary, this study demonstrates an important role of LXRs in protecting the liver against FB1-induced toxicity, suggesting an alternative mechanism not related to the inhibition of sphingolipid synthesis for mycotoxin toxicity.

Modulation of the expression of hepatic and renal genes encoding xenobiotic metabolizing enzymes by an aspalathin-enriched green rooibos (Aspalathus linearis) extract (GRE) was investigated in the liver and kidneys of F344 rats following dietary exposure of 28 d, as well as selected xenobiotic metabolizing genes in rat primary hepatocytes. In the liver, GRE upregulated genes (p

An aspalathin-enriched green rooibos (Aspalathus linearis) extract (GRE) was fed to male Fischer rats in two independent studies for 28 and 90 days. The average dietary total polyphenol (TP) intake was 756 and 627 mg Gallic acid equivalents (GAE)/kg body weight (bw)/day over 28 and 90 days, respectively, equaling human equivalent doses (HEDs) of 123 and 102 GAE mg/kg bw/day. Aspalathin intake of 295 mg/kg bw/day represents a HED of 48 mg/kg bw/day (90 day study). Consumption of GRE increased feed intake significantly (p &lt, 0.05) compared to the control after 90 days, but no effect on body and organ weight parameters was observed. GRE significantly (p &lt, 0.05) reduced serum total cholesterol and iron levels, whilst significantly (p &lt, 0.05) increasing alkaline phosphatase enzyme activity after 90 days. Endogenous antioxidant enzyme activity in the liver, i.e., catalase and superoxide dismutase activity, was not adversely affected. Glutathione reductase activity significantly (p &lt, 0.05) increased after 28 days, while glutathione (GSH) content was decreased after 90 days, suggesting an altered glutathione redox cycle. Quantitative Real Time polymerase chain reaction (PCR) analysis showed altered expression of certain antioxidant defense and oxidative stress related genes, indicative, among others, of an underlying oxidative stress related to changes in the GSH redox pathway and possible biliary dysfunction.

Populations consuming aflatoxin (AF) and fumonisin (FN)-contaminated foods may be at increased risk for hepatocellular carcinoma (HCC) and developmental disorders; consequently, development of intervention strategies to reduce AF/FN-induced liver disease and adverse health effects in humans could be very useful. Calcium montmorillonite clay (NovaSil) has been shown to absorb AF in vitro, in multiple animal models, as well as in human studies. In the present study, we aimed to evaluate whether uniform particle size NovaSil (UPSN) possessed an ability to modulate the co-carcinogenic potentials of aflatoxin B

The effect of bottle ageing on the antioxidant activity of Pinotage, Cabernet Sauvignon, Chardonnay and Chenin blanc wines, using the 2,2'-azino-di-(3-ethylbenzothialozine-sulphonic acid) radical cation (ABTS•+) and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH0) scavenging assays, was determined. Storage at 0°C, 15°C or 30°C for a period of 12 months resulted in a significant (p ≤ 0.05) decrease in both the total antioxidant activity (TAAAnTs and TAAoPPH) and the total phenol content of the wines. The antioxidant potency of the total phenols of most of the wines, which is a ratio of antioxidant activity to the total phenol content, also decreased. The total anthocyanins in the red wines decreased significantly (p ≤ 0.05) over 12 months except for storage at 0°C, while the flavanol content of the Pinotage, Cabernet Sauvignon and Chardonnay wines increased up to nine months storage with a subsequent decrease to 12 months. The flavonol content of all the wines decreased, while only minor changes in their hydroxycinnamate content were observed during the storage period. Understanding the complexity of these reactions may provide clues for stabilising especially red wines to preserve the antioxidant activity without losing the beneficial effects of colouring and flavour development during bottle ageing.

Among the many hurdles faced by developing countries, food safety with respect to mycotoxin contamination has frequently been side-lined with few countries having regulations and with poor enforcement where they do exist. Whereas commodity exporters may have the resources for engaging commercial accredited laboratories, the greatest challenge is found in rural, predominantly subsistence or smallholder farms, where conventional food surveillance is lacking. Rapid methods, designed for use in field conditions, where electricity is lacking or unreliable, can offer some solution to these problems. The World Food Programme's ‘Blue Box’ is an example of how technology can be adapted for these rural areas. The recent development of temperature stable aptamers and smart mobile phone technology may further enhance efforts to provide food safety in these areas.

Since the discovery of the fumonisins in 1988, extensive academic studies have generated much knowledge, including data on chemistry, biochemistry, toxicology, methods of analysis, natural occurrence in food supplies, fate during various processing procedures, and human and animal exposures. These mycotoxins have also been assessed by the International Agency for Research on Cancer (IARC) and have twice been the subject of risk assessments by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). The outcome of these investigations has been of a nature to alert risk managers to the necessity of controlling human exposure. However, the fumonisins occur mostly in maize, a world staple crop which is consumed in various communities at levels which can be as many as 100-fold different. Compounding the problem is the fact that maize is widely used as animal feed in many developed countries, whereas in Africa and some other developing countries, it is the primary food source. This contrast produces a problem for risk managers, partly solved at national level by the regulation of maximum tolerated levels (MTLs) applicable to individual countries. MTLs at an international level are currently under discussion at the Codex Committee on Contaminants in Food. The divergence in use and consumption and the fact that maize in various regions of the world can also vary greatly in contamination levels, leads to a dilemma for setting such MTLs, which would need to be low to protect the high maize consumers, but might then cause rejection of high amounts of the world supply. Higher MTLs, acceptable to maize exporters, would only protect the low maize consumers. This dilemma may only be solved by accepting that harmonizing regulations for raw maize is problematic and a more nuanced approach may be required.

Aflatoxins (AFs) and fumonisins (FBs) can co-contaminate foodstuffs and have been associated with hepatocellular and esophageal carcinomas in humans at high risk for exposure. One strategy to reduce exposure (and toxicity) from contaminated foodstuffs is the dietary inclusion of a montmorillonite clay (UPSN) that binds AFs and FBs in the gastrointestinal tract. In this study, the binding capacity of UPSN was evaluated for AFB1, FB1 and a combination thereof in Fischer 344 rats. Rats were pre-treated with different dietary levels of UPSN (0.25% or 2%) for 1 week. Rats were gavaged with a single dose of either 0.125 mg AFB1 or 25 mg FB1 per kg body weight and a combination thereof in the presence and absence of an aqueous solution of UPSN. The kinetics of mycotoxin excretion were monitored by analyzing serum AFB1 -albumin, urinary AF (AFM1) and FB1 biomarkers over a period of 72 h. UPSN decreased AFM1 excretion by 88-97%, indicating highly effective binding. FB1 excretion was reduced, to a lesser extent, ranging from 45% to 85%. When in combination, both AFB1 and FB1 binding occurred, but capacity was decreased by almost half. In the absence of UPSN, the combined AFB1 and FB1 treatment decreased the urinary biomarkers by 67% and 45% respectively, but increased levels of AFB1 -albumin, presumably by modulating its cytochrome metabolism. UPSN significantly reduced bioavailability of both AFB1 and FB1 when in combination; suggesting that it can be utilized to reduce levels below their respective thresholds for affecting adverse biological effects.

Maize is the predominant food source contaminated by fumonisins and this has particular health risks for communities consuming maize as a staple diet. The main biochemical effect of fumonisins is the inhibition of ceramide biosynthesis causing an increase in sphingoid bases and sphingoid base 1-phosphates and a depletion of the complex sphingolipids, thereby disrupting lipid metabolism and sphingolipid-mediated processes and signalling systems. Attempts to use the elevation of sphinganine as a human biomarker of fumonisin exposure have to date been unsuccessful. Consequently, recent research has focussed on developing a urinary exposure biomarker based on the measurement of the nonmetabolised toxin. In animals, fumonisins are poorly absorbed in the gut and are mostly excreted unmetabolised in faeces, with only a small percentage (0.25-2.0%) in urine. This appears to also be true in humans were fumonisin B1 (FB1) is detectable in urine soon after exposure, but in very small amounts relative to total intake. However, with modern sensitive and selective analytical methods such as liquid chromatography-tandem mass spectrometry, these low levels can be readily determined. The first study to show a positive correlation between consumption of maize and urinary FB1 was conducted in a Mexican population consuming tortillas as a staple food. Further validation of this relationship was achieved in a South African subsistence farming community with a positive correlation between urinary FB1 and fumonisin exposure, as assessed by food analysis and food intake data. The most recent developments are aimed at measuring multiple mycotoxin biomarkers in urine, including FB1. Current exposure studies in Guatemala are combining the urinary biomarker with measurement of sphinganine-1-phosphate in blood spots as a measure of biochemical effect. Thus, the urinary FB1 biomarker could contribute considerably in assessing the adverse health impact of fumonisin exposure.

Mycotoxin contamination of maize and maize-based food and feed products poses a health risk to humans and animals if not adequately controlled and managed. The current study investigates the effect of dry milling on the reduction of fumonisins (FB), deoxynivalenol (DON) and zearalenone (ZEA) in maize. Five composite samples, constructed to represent different mycotoxin contamination levels were degermed yielding degermed maize and the germ. The degermed maize was milled under laboratory conditions and four major milling fractions (SPECIAL, SUPER, semolina (SEM) and milling hominy feed) collected. The whole maize, degermed maize and total hominy feed (germ + milling hominy feed) were reconstructed to ensure homogenous samples for mycotoxin analyses. For comparison, commercial dry milling fractions (whole maize, SPECIAL, SUPER and total hominy feed), collected from three South African industrial mills, were analysed for the same mycotoxins and hence a more accurate assessment of the distribution between the different milling fractions. The distribution of the mycotoxins during the experimental dry milling of the degermed maize differs, with FB mainly concentrated in the SPECIAL, DON in the SEM whereas ZEA was equally distributed between the two milling fractions. Distribution of mycotoxins between the fractions obtained during commercial dry milling generally provided similar results with the total hominy feed containing the highest and the SUPER milling fractions the lowest mycotoxin levels although variations existed. Although milling is an effective way to reduce mycotoxins in maize, kernel characteristics and resultant fungal colonisation may impact on the distribution of specific mycotoxins among the different milling fractions. Differences in industrial dry milling practices and problems encountered in sampling bulk maize remain a large problem in assessing mycotoxin contamination in milling fractions intended for human consumption.

The chemopreventive properties of the herbal teas rooibos (Aspalathus linearis) and honeybush (Cyclopia spp.) have been demonstrated on mouse skin in vivo but the underlying mechanisms are not clear. The aim of the current study was to determine the anti-proliferative and pro-apoptotic activity of methanol and aqueous extracts of rooibos and two Cyclopia species in different skin cells, using green tea (Camellia sinensis) as a benchmark. Extracts were also characterised for their major individual polyphenols by high performance liquid chromatography and spectroscopically for the total polyphenol (TP) groups. The methanol extract of rooibos, containing higher levels of polyphenols than its aqueous extract, displayed similar activity to green tea as it selectively targeted premalignant cells by inhibiting cell proliferation at lower concentrations whilst inducing apoptosis via membrane depolarisation at higher concentrations. Specific roles of the major rooibos dihydrochalcones and flavanol/proanthocyanidin-type (FLAVA) compounds are likely to be involved. The aqueous extracts of the Cyclopia species were more active against cell proliferation and at inducing apoptosis which was associated with a higher FLAVA content and a reduced TP/FLAVA ratio. In contrast, their methanol extracts exhibited a cytoprotective effect against apoptosis which was related to their monomeric xanthone and flavanone content. The underlying chemopreventive properties of green tea and the herbal teas appear to be associated with diverse and complex monomeric/polymeric polyphenolic cell interactions.

Background Sutherlandia frutescens (L.) R. Br is endemic to Southern Africa where it has been traditionally used for cancer and diabetes. In recent times it has been marketed for its reputed (but not proven) anticancer, antidiabetic and anti-HIV properties. Little is known about the mutagenic and antimutagenic potential of extracts and common marker compounds of Sutherlandia frutescens. Therefore this study aimed to investigate the putative efficacy and possible long-term adverse effects of using this herb. Methods Ethylacetate (EA) and 50% Methanol (MeOH) extracts were screened for mutagenic and antimutagenic activity using the Ames assay utilising TA97a, TA98, TA100 and TA102 in the presence and absence of metabolic activation. Four compounds, L-arginine, L-canavanine, GABA and D-pinitol known to occur in sutherlandia were also included. The total polyphenolic content of the both extracts was determined using the Folin-Ciocalteau method and FRAP and ABTS were used to determine the anti-oxidant potential of the extracts. Results The extracts and the standards did not show any cytotoxicity except in TA97a. The EA extract exhibited antimutagenicity against all the bacterial strains at all concentrations tested. The MeOH extract showed both pro-mutagenic and antimutagenic activities with 2-acetamidofluorene and aflatoxin B1 in the presence of metabolic activation of TA98 and TA100, respectively. All compounds, except L-canavanine exhibited antimutagenic activity against all strains. L-canavanine, on the other hand showed co-mutagenicity with 9-aminoacridine on TA97a, at all test concentrations. The extracts and pure compounds exhibited their antimutagenic activity in a dose response manner. L-arginine and GABA showed an some antimutagenic response. EA extract had three times the total phenolic content (12.56 μg GE / mg) observed in the MeOH extract. There was correlation between total phenolic content, antioxidant potential and antimutagenicity. Conclusion Both extracts exhibited a protective effect, with the EA extract exhibiting greater potency. L-canavanine acted as a co-mutagen in a dose response manner without metabolic activation. It is suggested that the EA extract be priotized for future development work as it showed a better risk profile and activity.

Dietary co-exposure to aflatoxin B1 (AFB1) and fumonisin B1 (FB1) and their interaction on hepatocellular carcinogenesis is of particular concern in toxicology and public health. In this study we evaluated the liver preneoplastic effects of single and sequential dietary exposure to AFB1 and FB1 in the F344 rat carcinogenesis model. Serum biochemical alterations, liver histopathological changes, and the formation of liver glutathione S transferase positive (GST-P+) foci were the major outcome parameters examined. Compared to the AFB1-only treatment, the FB1-only treatment induced less dysplasia, and more apoptosis and mitoses. Sequential AFB1 and FB1 treatment lead to increased numbers of dysplasia, apoptosis and foci of altered hepatocytes, as compared to either mycotoxin treatment alone. More importantly, sequential exposure to AFB1 and FB1 synergistically increased the numbers of liver GTP-P+ foci by approximately 7.3-and 12.9-fold and increased the mean sizes of GST-P+ foci by 6- and 7.5-fold, respectively, as compared to AFB1- or FB1-only treatment groups. In addition, liver ALT and AST levels were significantly increased after sequential treatment as compared to single treatment groups. The results demonstrate the interactive effect of dietary AFB1 and FB1 in inducing liver GST-P+ foci formation and provide information to model future intervention studies.

Mangiferin displays an extensive spectrum of pharmacological properties, including antioxidant activity. Its phase II metabolism in the presence of Aroclor 1254-induced and un-induced microsomal and cytosolic fractions from rat liver and the antioxidant potency of the glucuronidated conjugates were investigated. Mangiferin was not a substrate for the cytosolic sulphotransferases. Glucuronidation led to the formation of two monoglucuronidated metabolites of mangiferin and a monoglucuronidated metabolite of homomangiferin (a minor constituent of the mangiferin standard). Deconjugation utilising glucuronidase resulted in the disappearance of the metabolites, with the concomitant formation of the two parent compounds. Considering steric hinderance caused by the C-2 glucosyl moiety and the relative acidity of the xanthone OH groups, the 6-OH of mangiferin and, to a lesser degree the 7-OH, are likely to be the primary glucuronidation targets. The ferric iron reducing ability of the glucuronidated reaction mixture was reduced, while the free radical scavenging abilities of mangiferin, utilising on-line post-column HPLC-DAD-DPPH and HPLC-DAD-ABTS + assays, were eliminated, providing further evidence that the catechol arrangement at C-6 and C-7 was the preferred site of conjugation. This paper provides the first evidence that the glucuronidated metabolites of mangiferin resulted in a loss in free radical scavenging and ferric iron reducing ability.

The steroid hormone output of the adrenal gland is crucial in the maintenance of hormonal homeostasis, with hormonal imbalances being associated with numerous clinical conditions which include, amongst others, hypertension, metabolic syndrome, cardiovascular disease, insulin resistance and type 2 diabetes. Aspalathus linearis (Rooibos), which has been reported to aid stress-related symptoms linked to metabolic diseases, contains a wide spectrum of bioactive phenolic compounds of which aspalathin is unique. In this study the inhibitory effects of Rooibos and the dihydrochalcones, aspalathin and nothofagin, were investigated on adrenal steroidogenesis. The activities of both cytochrome P450 17α-hydroxylase/17,20 lyase and cytochrome P450 21-hydroxylase were significantly inhibited in COS-1 cells. In order to study the effect of these compounds in H295R cells, a human adrenal carcinoma cell line, a novel UPLC-MS/MS method was developed for the detection and quantification of twenty-one steroid metabolites using a single chromatographic separation. Under both basal and forskolin-stimulated conditions, the total amount of steroids produced in H295R cells significantly decreased in the presence of Rooibos, aspalathin and nothofagin. Under stimulated conditions, Rooibos decreased the total steroid output 4-fold and resulted in a significant reduction of aldosterone and cortisol precursors. Dehydroepiandrosterone-sulfate levels were unchanged, while the levels of androstenedione (A4) and 11β-hydroxyandrostenedione (11βOH-A4) were inhibited 5.5 and 2.3-fold, respectively. Quantification of 11βOH-A4 showed this metabolite to be a major product of steroidogenesis in H295R cells and we confirm, for the first time, that this steroid metabolite is the product of the hydroxylation of A4 by human cytochrome P450 11β-hydroxylase. Taken together our results demonstrate that Rooibos, aspalathin and nothofagin influence steroid hormone biosynthesis and the flux through the mineralocorticoid, glucocorticoid and androgen pathways, thus possibly contributing to the alleviation of negative effects arising from elevated glucocorticoid levels.

The British mycologist, I.B. Pole-Evans, was appointed as the first South African government mycologist in 1905 following the Anglo-Boer War (1899-1902). The Onderstepoort Veterinary Research Institute was founded in 1908 with the Swiss veterinarian, Arnold Theiler, as the first director. Thus, the stage was set for the commencement of mycotoxicological research when the Union of South Africa came into being in 1910. The first accounts of this pioneering research appeared in the 'Seventh and eight reports of the Director of Veterinary Research, Union of South Africa. 1918' in which D.T. Mitchell reported on the experimental reproduction of the neurotoxic syndrome, diplodiosis, in cattle with pure cultures of Stenocarpella maydis (= Diplodia zea) isolated by P.A. Van der Bijl and grown on sterile maize kernels. This is the first report of the experimental reproduction of a veterinary mycotoxicosis with a pure culture of a fungus in South Africa and possibly in the world. This seminal research was followed by a great deal of multidisciplinary research on veterinary mycotoxicoses as well as human syndromes in which fungal toxins are suspected to be involved, taxonomy of mycotoxigenic fungi and chemistry of mycotoxins in South Africa. The mycotoxicoses studied in South Africa include the following (more or less in chronological order): diplodiosis, Paspalum staggers, aflatoxicosis, human hepatocellular carcinoma, ochratoxicosis, lupinosis, facial eczema, tremorgenic mycotoxicosis, hyperoestrogenism, stachybotryotoxicosis, ergotism, leukoencephalomalacia and human oesophageal cancer. A major breakthrough in mycotoxicological research was made in South Africa in 1988 with the isolation and chemical characterisation of the carcinogenic fumonisins produced by Fusarium verticillioides in maize. Current research at the PROMEC Unit of the South African Medical Research Council on the risk assessment of fumonisins and intervention methods to reduce fumonisin intake by rural populations on a maize staple diet is highlighted. This paper concludes with a selected list of mycotoxicological publications by South African mycologists/plant pathologists, veterinarians and chemists/biochemists.

Background: The consumption of maize highly contaminated with carcinogenic fumonisins has been linked to high oesophageal cancer rates. The aim of this study was to validate a urinary fumonisin B1 (UFB1) biomarker as a measure of fumonisin exposure and to investigate the reduction in exposure following a simple and culturally acceptable intervention. Methods: At baseline home-grown maize, maize-based porridge, and first-void urine samples were collected from female participants (n = 22), following their traditional food practices in Centane, South Africa. During intervention the participants were trained to recognize and remove visibly infected kernels, and to wash the remaining kernels. Participants consumed the porridge prepared from the sorted and washed maize on each day of the two-day intervention. Porridge, maize, and urine samples were collected for FB1 analyses. Results: The geometric mean (95% confidence interval) for FB1 exposure based on porridge (dry weight) consumption at baseline and following intervention was 4.84 (2.87–8.14) and 1.87 (1.40–2.51) μg FB1/kg body weight/day, respectively, (62% reduction, P < 0.05). UFB1C, UFB1 normalized for creatinine, was reduced from 470 (295–750) at baseline to 279 (202–386) pg/mg creatinine following intervention (41% reduction, P = 0.06). The UFB1C biomarker was positively correlated with FB1 intake at the individual level (r = 0.4972, P < 0.01). Urinary excretion of FB1 was estimated to be 0.075% (0.054%–0.104%) of the FB1 intake. Conclusion: UFB1 reflects individual FB1 exposure and thus represents a valuable biomarker for future fumonisin risk assessment. Impact: The simple intervention method, hand sorting and washing, could positively impact on food safety and health in communities exposed to fumonisins. Cancer Epidemiol Biomarkers Prev; 20(3); 483–9. ©2011 AACR.

Subsistence farming communities with low socio-economic status reliant on a mono cereal maize diet are exposed to fumonisin levels that exceed the provisional maximum tolerable daily intake of 2 μg kg −1 body weight day −1 recommended by the Joint FAO/WHO Expert Committee on Food Additives. In the rural Centane magisterial district, Eastern Cape Province, South Africa, it is customary during food preparation to sort visibly infected maize kernels from good maize kernels and to wash the good kernels prior to cooking. However, this customary practice seems not to sufficiently reduce the fumonisin levels. This is the first study to optimise the reduction of fumonisin mycotoxins in home-grown maize based on customary methods of a rural population under laboratory-controlled conditions. Maize obtained from subsistence farmers was analysed for the major naturally occurring fumonisins (FB 1 , FB 2 and FB 3 ) by fluorescence HPLC. Large variations were observed in the unsorted and the experimental maize batches attributable to the non-homogeneous distribution of fumonisin contamination in maize kernels. Optimised hand-sorting of maize kernels by removing the visibly infected/damaged kernels (fumonisins, 53.7 ± 15.0 mg kg −1 , 2.5% by weight) reduced the mean fumonisins from 2.32 ± 1.16 mg kg −1 to 0.68 ± 0.42 mg kg −1 . Hand washing of the sorted good maize kernels for a period of 10 min at 25 °C resulted in optimal reduction with no additional improvement for wash periods up to 15 h. The laboratory optimised sorting reduced the fumonisins by 71 ± 18% and an additional 13 ± 12% with the 10 min wash. Based on these results and on local practices and practicalities the protocol that would be recommended to subsistence farmers consists of the removal of the infected/damaged kernels from the maize followed by a 10 min ambient temperature water wash.

Fusarium verticillioides causes several animal diseases and the contamination maize suggests that it could adversely affect human health. The fumonisin B mycotoxins were characterized from the fungal culture material and shown to be the causative principle responsible for the major mycotoxicological effects of the fungus in experimental and farm animals. The main focus was on the toxicological effects in rats and mice, the outcome of which played an important role in setting risk assessment parameters for exposure of the fumonisins to humans. The International Agency for Research on Cancer characterized the fumonisins as Group 2B carcinogens. Several controversial findings regarding the toxicological effects of the culture material of the fungus, the genotoxicity and carcinogenicity of pure fumonisin B(1) (FB(1)) in rats have been reported that should be clarified prior to assessing the risk in humans. The underlying differences between the diets with the high protein levels are likely to sensitize the kidneys to FB(1)-induced toxic and carcinogenic effects. Several other dietary factors, such as plant extracts (antioxidants) and dietary Fe, could either stimulate or inhibit cancer induction of FB(1), which complicates the comparison of toxicological effects in experimental animals. Cognisance should be taken of the modulating role of dietary constituents as it will determine the outcome of toxicological assays and determine the threshold of an adverse effect in a specific target organ to be used in determining risk assessment parameters.

A validated culturally specific dietary assessment method was used to determine the habitual maize intakes of black Xhosa-speaking Africans living in the Centane region of the Eastern Cape Province to assess their exposure to the carcinogenic fumonisin mycotoxins. The mean total dry weight maize intakes of home-grown, commercial or combined (both maize sources) were 474, 344, 462 g day(-1), respectively. When considering the total mean levels of fumonisin in home-grown maize (1142 microg kg(-1)) and commercial maize (222 microg kg(-1)), the probable daily intakes (PDI's), expressed as microg kg(-1) body weight day(-1) were 12.1 (95%CI: 0.3-4926.5) and 1.3 (95%CI: 1.0-1.8) for men and 6.7 (95%CI: 1.0-457.8) and 1.1 (95%CI: 0.9-1.3) for women, consuming home-grown and commercial maize, respectively. Based on the different maize-based beer drinking frequencies the PDI's varied between 6.9 and 12.0 microg kg(-1)/drinking event. Depending on the maize intake patterns an exposure "window" exists where fumonisin exposure is below the recommended group provisional maximum tolerable daily intake (PMTDI) for fumonisins of 2 microg kg(-1)bw day(-1). The assessment of fumonisin exposure and development of preventative strategies depend, not only the accurate determination of total fumonisin levels in maize, but also on the distinct dietary patterns of a specific population.

Aims: To investigate the antifungal activity of aqueous and organic extracts of four weedy plant species viz. Tagetes minuta, Lippia javanica, Amaranthus spinosus and Vigna unguiculata against isolates of four agriculturally important fungi, i.e. Fusarium verticillioides, F. proliferatum, Aspergillus flavus and A. parasiticus. Methods and Results: Dried powdered aerial parts of the plants were extracted sequentially with hexane, dichloromethane, methanol and water and tested for activity using a serial microdilution assay. Results were read every day over 120 h. All extracts except for the water extracts showed growth inhibitory activity against most isolates of the Fusarium spp. The most active were the methanol and hexane extracts of V. unguiculata and A. spinosus with minimum inhibitory concentration (MIC) values of

Cancer incidence rates and patterns are reported for a rural population, living in the Eastern Cape Province of South Africa for the period 1998-2002. The population-based cancer registry has operated for 20 years, using both active and passive methods for case finding, through collaborations with 19 health facilities: 11 district hospitals, 7 referral hospitals and 1 regional laboratory. The age standardized incidence rates for all cancers were 73.1 per 100,000 in males and 64.1 per 100,000 in females. The leading top 5 cancers for males were oesophagus (32.7 per 100,000), lung (5.8 per 100,000), prostate (4.4 per 100,000), liver (4.4 per 100,000) and larynx (2.5 per 100,000) whereas for females they were cervix (21.7 per 100,000), oesophagus (20.2 per 100,000), breast (7.5 per 100,000), ovary (0.9 per 100,000) and liver (0.9 per 100,000). The incidence of Kaposi sarcoma was low, and higher for males (1.6 per 100,000) than females (0.3 per 100,000). Lung cancer in both males and females was relatively low compared to the high incidence of oesophagus cancer.

Aspalathin (2',3,4,4',6'-pentahydroxy-3'-C-beta-d-glucopyranosyldihydrochalcone) is the major flavonoid present in the South African herbal tea rooibos. In vitro metabolism of aspalathin and a structural analogue nothofagin, lacking the A ring catechol group, was investigated by monitoring the formation of glucuronyl and sulfate conjugates using Aroclor 1254 induced and uninduced rat liver microsomal and cytosolic subcellular fractions. Following glucuronidation of both aspalathin and nothofagin, HPLC-DAD, LC-MS, and LC-MS/MS analyses indicated the presence of two metabolites: one major and one minor. Only one aspalathin metabolite was obtained after sulfation, while no metabolites were observed for nothofagin. Two likely sites of conjugation for aspalathin are 4-OH or 3-OH on the A-ring. For nothofagin, the 4-OH (A-ring) and 6'-OH (B-ring) seem to be involved. The glucuronyl conjugates of aspalathin lack any radical scavenging properties in online postcolumn DPPH radical and ABTS radical cation assays. Deconjugation assays utilizing glucuronidase and sulfatase resulted in the disappearance of the metabolites, with the concomitant formation of the unconjugated form in the case of the glucuronidated product. The balance between conjugated and unconjugated forms of aspalathin could have important implications regarding its role in affecting oxidative status in intra- and extracellular environments in vivo.

Altered membrane integrity in hepatocellular carcinoma (HCC) tissue was indicated by an elevation in cholesterol and significant decrease in phosphatidylcholine (PC). The resultant decreased phosphatidylcholine/phosphatidylethanolamine (PC/PE) and increased cholesterol/phospholipid ratios are associated with decreased fluidity in the carcinoma tissue. The lower PC was associated with a decrease in the quantitative levels of the saturated (C16:0, C18:0), omega6 (C18:2, C20:4) and omega3 (C22:5, C22:6) fatty acids (FAs), resulting in reduced long-chain polyunsaturated fatty acids (LCPUFAs), total PUFA and an increase in omega6/omega3 FA ratio. In PE, the saturated and omega3 (C22:5, C22:6) FAs were reduced while the total omega6 FA level was not affected, leading to an increased omega6/omega3 FA ratio. Increased levels of C18:1omega9, C20:2omega6 and reduction of 22:6omega3 in PC and PE suggest a dysfunctional delta-6 desaturase. The reduced PC/PE ratio resulted in a decreased C20:4omega6 (PC/PE) ratio, implying a shift towards synthesis of the 2-series eicosanoids. Lipid peroxidation was reduced in both hepatitis B negative (HBV(-)) and positive (HBV(+)) HCC tissues. Glutathione (GSH) was decreased in HCC while HBV had no effect, suggesting an impairment of the GSH redox cycle. In contrast HBV infection enhanced GSH in the surrounding tissue possibly to counter oxidative stress as indicated by the increased level of conjugated dienes. Apart from the reduced LCPUFA, the low level of lipid peroxidation in the carcinoma tissue was associated with increased superoxide dismutase and glutathione peroxidase activity. The disruption of the redox balance, resulting in increased cellular antioxidant capacity, could create an environment for resistance to oxidative stress in the carcinoma tissue. Alterations in membrane cholesterol, phospholipids, FA parameters, C20:4omega6 membrane distribution and low lipid peroxidation are likely to be important determinants underlying the selective growth advantage of HCC cells.

The chemoprotective properties of unfermented and fermented rooibos (Aspalathus linearis) and honeybush (Cyclopia intermedia) herbal teas, and green and black teas (Camellia sinensis) were investigated against fumonisin B1 (FB1) promotion in rat liver utilizing diethylnitrosamine (DEN) as cancer initiator. The various teas differently affected the clinical chemical parameters associated with liver and kidney damage associated with FB1 suggesting specific FB1/iron/polyphenolic interactions. Green tea enhanced (P0.05) the FB1-induced reduction of the oxygen radical absorbance capacity, while fermented herbal teas and unfermented honeybush significantly (P0.05) decreased FB1-induced lipid peroxidation in the liver. The teas exhibited varying effects on FB1-induced changes in the activities of catalase, glutathione peroxidase (GPx) glutathione reductase (GR) as well as the glutathione (GSH) status. Unfermented rooibos and honeybush significantly (P0.05) to marginally (P0.1) reduced the total number of foci (10microm), respectively, while all the teas reduced the relative amount of the larger foci. Fermentation seems to reduce the protective effect of the herbal teas. Differences in the major polyphenolic components and certain FB1/polyphenolic/tissue interactions may explain the varying effects of the different teas on the oxidative parameters, hepatotoxic effects and cancer promotion in rat liver.

The in vitro antioxidant activity of aqueous extracts prepared from four Cyclopia spp. (unfermented and fermented) was assessed using radical (ABTS *+) scavenging, ferric ion reduction, and inhibition of Fe2+-induced microsomal lipid peroxidation as criteria. Aqueous extracts of unfermented and fermented Aspalathus linearis (rooibos) and Camellia sinensis teas (green, oolong, and black) were included as reference samples. Qualitative and quantitative differences in phenolic composition were demonstrated for the Cyclopia spp. The xanthone glycoside, a.k.a. mangiferin, was the major monomeric polyphenol present in the Cyclopia extracts, with both unfermented and fermented C. genistoides extracts containing the highest quantities. Fermentation resulted in a significant reduction in extract yields and their total polyphenolic and individual polyphenol contents. Unfermented plant material should preferentially be used for preparation of extracts, as fermentation significantly ( P0.05) lowered antioxidant activity of all species, except in the case of C. genistoides, where the ability to inhibit lipid peroxidation was not affected. Unfermented plant material also retained the highest concentration of mangiferin. Overall, extracts of unfermented Cyclopia were either of similar or lower antioxidant activity as compared to the other teas. However, the presence of high levels of mangiferin merits the use of Cyclopia spp. and, in particular, C. genistoides, as an alternative herbal tea and potential dietary supplement.

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.

1 H and 13 C NMR spectroscopy of both fumonisin B 3 and B 4 , as well as high-performance liquid chromatography (HPLC) analysis of samples of fumonisin B 3 used as standards, showed in each case the presence of two stereoisomers, which could not be separated by preparative chromatography. The 2,3-anti relative configuration for the two minor stereoisomers of fumonisin B 3 and B 4 was deduced from the NMR data, and their 2S,3R absolute configurations were established by application of Mosher's method using the fumonisin B 3 sample. Samples of fumonisin B 3 and B 4 can contain between 10 and 40% of fumonisin B compounds of the 3-epi series. The 3-epi-FB 3 , determined by HPLC with fluorescence detection of the o-phthaldialdehyde derivative and confirmed by liquid chromatography-tandem mass spectrometry, was found to occur naturally in a range of maize samples at levels much lower than FB 3 (

Antimutagenic activity of aqueous extracts of the South African herbal teas, Aspalathus linearis (rooibos) and Cyclopia spp. (honeybush) was compared with that of Camellia sinensis (black, oolong and green) teas in the Salmonella mutagenicity assay using aflatoxin B(1) (AFB(1)) and 2-acetylaminofluorene (2-AAF) as mutagens. The present study presents the first investigation on antimutagenic properties of C. subternata, C. genistoides and C. sessiliflora. The herbal teas demonstrated protection against both mutagens in the presence of metabolic activation, with the exception of "unfermented" (green/unoxidised) C. genistoides against 2-AAF, which either protected or enhanced mutagenesis depending on the concentration. Antimutagenic activity of "fermented" (oxidised) rooibos was significantly (P0.05) less than that of Camellia sinensis teas against AFB(1), while for 2-AAF it was less (P0.05) than that of black tea and similar (P0.05) to that of oolong and green teas. Antimutagenic activity of unfermented C. intermedia and C. subternata exhibited a similar protection as fermented rooibos against AFB(1). Against 2-AAF, fermented rooibos exhibited similar protective properties than unfermented C. intermedia and C. sessiliflora. Unfermented rooibos was less effective than the C. sinensis teas and fermented rooibos, but had similar (P0.05) antimutagenicity to that of fermented C. sessiliflora against AFB(1) and fermented C. subternata against 2-AAF. Fermented C. intermedia and C. genistoides exhibited the lowest protective effect against 2-AAF, while fermented C. intermedia exhibited the lowest protection when utilising AFB(1) as mutagen. Aspalathin and mangiferin, major polyphenols in rooibos and Cyclopia spp., respectively, exhibited weak to moderate protective effects when compared to the major green tea catechin, (-)epigallocatechin gallate (EGCG). Antimutagenic activity of selected herbal tea phenolic compounds indicated that they contribute towards (i) observed antimutagenic activity of the aqueous extracts against both mutagens and (ii) enhancement of the mutagenicity of 2-AAF by unfermented C. genistoides. Antimutagenic activity of the South African herbal teas was mutagen-specific, affected by fermentation and plant material, presumably due to changes and variation in phenolic composition.

Unfermented rooibos tea is known to contain higher levels of total polyphenols and flavonoids than its fermented counterpart, making it the obvious choice for the preparation of flavonoid-enriched fractions. Evaluation of aqueous extracts and crude polyphenolic fractions of unfermented and fermented rooibos showed anti- and/or pro-oxidant activities, using a linoleic acid-Tween-buffer emulsion for lipid peroxidation and the deoxyribose degradation assay, based on a Fenton reaction model system containing FeCl3-EDTA and H2O2 for the generation of hydroxyl radicals. Except for the ethyl acetate fraction, with the highest total polyphenol (TP) content and offering the least protection presumably due to pro-oxidant activity, the inhibition of lipid peroxidation by the samples correlated moderately with their TP content in a linear relationship (r = 0.896, P0.01). Using the deoxyribose degradation assay, the pro-oxidant activity of the aqueous extracts and their crude polymeric fractions (0.1 mg/mL in the reaction mixture) was linear with respect to their dihydrochalcone (aspalathin and nothofagin) (r = 0.977, P = 0.023) and flavonoid (r = 0.971, P = 0.029) content. Pro-oxidant activity was demonstrated for pure aspalathin. Using the same assay, but with ascorbate added to regenerate Fe3+ to Fe2+, the aqueous extract and crude polymeric fraction of fermented rooibos displayed hydroxyl radical scavenging activity. Fermentation (i.e., oxidation) of rooibos decreased the pro-oxidant activity of aqueous extracts, which was contributed to a decrease in their dihydrochalcone content. The in vitro pro-oxidant activity displayed by flavonoid-enriched fractions of rooibos demonstrates that one must be aware of the potential adverse biological properties of potent antioxidant extracts utilized as dietary supplements.