Aflatoxins in Food

Initially, we set out to explore the presence of aflatoxins in food. However, as we dove deeper into the subject, we stumbled upon some unexpected discoveries. Hence this article ventures beyond aflatoxins in food. Here we we explore European safety limits, safety limits in humans, average daily intakes based on diet type and scientifically proven methods to reduce and eliminate aflatoxins.

Summary

• 25% of all food crops world wide are contaminated with mycotoxins.
• Out of 300 identified mycotoxins, aflatoxins are thought to be some of the most potent mutagenic and carcinogenic substances known.
• There are no official safety limits in humans. Instead, a “provisional” safety limit is used.
• Provisional safety limit has several problems because it does not take into consideration: 1. additive or synergistic effects for two or more mycotoxin and 2. toddlers, infants and children.
• Peanuts and cereals from Asia (China) and South America (Argentina) are the most frequently contaminated sources of aflatoxin B1.
• On average, one tablespoon of peanut butter can give you 57% of your daily provisional safety limit for aflatoxin B1.
• On a daily basis, vegetarians and vegans consume much higher levels of aflatoxin B1.
• Toddlers, infants and children following vegetarian and vegan diets exceed aflatoxin B1 limits by a magnitude of 4 or more.
• Soaking, washing and pressure cooking can reduce or eliminate aflatoxin B1 in food.

Contents

1. Mycotoxins
2. Mycotoxins a World Wide problem
3. Mycotoxin Contamination
4. Mycotoxin Health Effects
5. Aflatoxin B1 Safety Limits in Humans
6. EU Limits for Aflatoxins in Food
7. Worst case Aflatoxin B1 Estimates in EU Food
8. Estimating Aflatoxin B1 in Various Diets
9. Aflatoxin B1 estimates discussed
10. Special Consideration for Children
11. Reducing Aflatoxins

Mycotoxins

Mycotoxins are naturally occurring substances mainly produced by three fungal genera: Aspergillus, Fusarium, and Penicillium. Out of the 300 mycotoxins identified, eight are consistently found in the food chain (Table. 1) and pose serious health risks worldwide ¹.

Aflatoxins are a group of mycotoxins split into different groups. These comprise aflatoxin B1, B2, G1, G2 and M1. Amongst these five toxins, aflatoxin B1 (AFB1) is the most common and most toxic. The International Agency for Research on Cancer (IARC) has classified aflatoxins as group 1 carcinogens for humans ².

Mycotoxins a World Wide problem

Two decades ago, the Food and Agriculture Organisation (FAO) estimated 25% of all food crops world wide to be contaminated by mycotoxins. Since then, there has been a rise in extensive standards, legislations and good agricultural practices. All of which are enforced at a world, European Union (EU) and national levels ⁴. However, despite the ongoing battle to reduce mycotoxins, its prevalence remains the same ⁵.

At present, an estimated 4.5 billion people are chronically exposed to largely uncontrolled levels of mycotoxins ⁶. Although such occurrences are less frequent in developed countries, exposure is chronic. There are detectable levels of mycotoxins in 60 to 80% of foods ⁷. Recent estimations have predicted further increases in levels of DON and AFB1 contamination in European cereals ⁸. A result attributed to the effects of global warming.

Mycotoxin Contamination

Regardless of agricultural, storage and processing practices implemented, mycotoxin contamination is considered a persistent and unpredictable problem ⁹. Their ability to withstand heat, physical and chemical treatments make them especially difficult to control. Certain factors encourage even higher levels of mycotoxins in food. These include climatic conditions, insect infestation, drought before harvest and sub optimal storage practices.

Mycotoxin Health Effects

Mycotoxins exert a multitude of toxic effects on animals and humans. Exposure ranges from acute (high mycotoxin doses, short-term exposure) to chronic (lower mycotoxin doses, long-term exposure). The main target organs are the liver, gut, lungs, kidneys, central nervous system and the immune system. Once a mycotoxin reaches its target(s), it may exert haemorrhagic, dermatotoxic, hepatotoxic, nephrotoxic, neurotoxic, oestrogenic, teratogenic, immunosuppressive, mutagenic and/or carcinogenic effects ^{10 11}.

An individual’s susceptibility depends on several factors, including gender, age, diet, overall health, dose, type of mycotoxin, and length of exposure. Another layer of complexity is added in light of the evidence that mycotoxins have additive and/or synergistic effects (that is they can amplify the effects of each others) ^{12 13 14 15 16}. And since food contains a mixture of mycotoxins, determining their safety limits and mechanisms of toxicity is challenging.

Aflatoxin B1 Safety Limits in Humans

Although aflatoxins “are some of the most potent mutagenic and carcinogenic substances known”, there are no official safety limits for AFB1 in humans ¹⁷. A threshold at which AFB1  shows no effects has not been determined. And without a threshold, it’s not possible to establish Tolerable Daily Intake (TDI) – the daily intake of a substance established to be safe for long term human consumption. Instead, AFB1 ‘safety limit’ is based on a non official figure – the ‘provisional maximal’ TDI (PMTDI). The PMTDI for AFB1 is 1 ng/kg^-1 bw/day^-1 (1 nanogram per kilogram of body weight per day) ¹⁸. Hence a person weighing 70 kg would have a PMTDI of 70 ng AFB1. 

It’s important to note that not all bodies agree with this provisional safety limit. The European Union Scientific Committee for Food (SCF) and Joint Expert Committee on Food Additives (JECFA) concluded that exposure to as little as 1 ng/kg^-1 bw/day^-1 or even less, may contribute to risk of liver cancer ^{19 20}. There are two additional concerns with AFB1 PMTDI: 1. it does not take into consideration health risks due to additive and/or synergistic effects for two or more mycotoxins and 2. it does not take into consideration toddlers, infants and children who have an unfavourable ratio of food intake per body weight. Hence the general consensus is to reduce AFB1 exposure as low as possible.

EU Limits for Aflatoxins in Food

The European Union has set its own regulations for levels of aflatoxins in food (Table. 2). The maximal tolerated levels for total aflatoxins (sum of B1, B2, G1, G2) range between 4-15 µg/kg (micrograms per kilogram). While maximal AFB1 levels range from 2-12 µg/kg. 

Groundnuts (e,g., peanuts) and cereals (e.g., rice and maize) are some of the most frequently contaminated sources of AFB1. The maximum limit for these foods is 2 µg/kg. In contrast, the limit for tree nuts (e.g., almonds, pistachios and apricot kernels) is 4 times greater (8µg/kg). 

Worst case Aflatoxin B1 estimates in EU food

In order to visualise the EU limits, we produced a “worst case” exposure plot for a 70kg person (Figure.1). One table spoon of dried spice (e.g., chilli, ginger, turmeric, black pepper) may contain 36% (25ng) of the daily limit for AFB1. While 15g of dried fruits (e.g., figs, dates and raisins) 43% (30ng).

The most frequently contaminated sources of AFB1 are peanuts, cereals (e.g., oats and rice) and cereal products (e.g., breakfast cereal and bread). These foods may contain the following amounts of AFB1 PMTDI: peanut butter 57% (40ng), cooked oats 229% (160ng), cooked rice 314% (220ng), breakfast cereal 200% (140ng), and bread 71% (50ng).

Although almonds are less frequently contaminated than peanuts and cereals, it’s possible for one tablespoon of almond butter to exceed the daily limit by 285% (200ng).

Consuming these foods in the stated quantities would equal to ~964 calories and exceed the AFB1 PMTDI by 1250% (875ng). Adjusting food to equal 2000 calories yields a total of 1800ng of AFB1 (2571% of PMTDI) (approximately 0.9ng AFB1 per calorie).

These worst case estimates fully comply with current EU limits for AFB1.

Estimating Aflatoxin B1 Various Diets

As of yet, no studies have measured AFB1 occurrence in alternative diets (Keto, Paleo, Vegetarian, Vegan, Whole Food Plant Based (WFPB) and Vegan keto). Given the nature of AFB1 toxicity, the worst case estimates, and the rise in popularity of alternative diets, we decided to investigate this matter further.

Mean AFB1 concentration in European foods was taken from EFSA CONTAM Panel and averaged (Table. 3). The panel lists a total of 57,983 samples, taken from 26 European countries between 2013-2018. Most of the samples came from Germany and Netherlands (27% for each), followed by France (11%). The median limit of quantification (LOQ) was 1 µg/kg. This is the the lowest analyte concentration that can be quantitatively detected with a given accuracy and precision (usually 95%).

Out of 18 food categories, all were contaminated with AFB1. The highest mean AFB1 concentration was for the food category ‘legumes, nuts and oilseeds’, and for the food category ‘herbs, spices and condiments’. In contrast, meat and meat products, fish and other seafood, milk and dairy products, and infant food contained the lowest.

Using the data above we calculated the mean intake of AFB1 for various diet groups (Figure. 2.). The two groups estimated to have the highest concentration were Whole Food Plant Based (WFPB) (0.8 ng/kg^-1 bw/day^-1) and vegan keto (0.9 ng/kg^-1 bw/day^-1).

Aflatoxin B1 estimates discussed

Our estimates for the standard diet were higher than those reported in the literature ¹⁷. This is due to the nature of our crude estimates (we assumed equal intakes from all food categories). In reality, the standard diet has higher intakes of animal products, processed foods (lower in AFB1) and less wholesome plant foods (higher in AFB1).

According to SCF and JECFA, our WFPB and vegan keto groups are at risk for AFB1-induced health effects, even though estimated AFB1 exposure was less than AFB1 PMTDI. The risk is likely to be even greater due to concomitant presence of mycotoxins in food which can lead to additive or even synergistic effects. Animal models and cell line studies have shown AFB1 synergy with OTA ²⁵, DON and fumonisin B1 ²⁶, and various other mycotoxins ²⁷. This toxicological risk is completely ignored by AFB1 PMTDI and is especially of concern given the chronic daily exposure to aflatoxins.

No AFB1 exposure data was available to compare our keto, paleo, WFPB and vegan keto groups. However, total aflatoxin occurrence data was available for vegetarian and vegan groups. Given that AFB1 is the dominant aflatoxin, it is reasonable to assume that our estimates are lower than what has been previously reported ²⁸. This suggests vegetarians and especially vegans could be at high risk as well.

We expect to see substantial fluctuations of AFB1 concentrations during warmer and more humid months. In addition, we predict higher AFB1 exposure in EU countries with higher percentage of imported foods from Asia and South America. For example, the United Kingdom (UK) is one of the biggest importers of food in the world ²⁹. A large proportion of problematic foods (e.g., peanuts, rice and nuts) are imported from China ³⁰, Argentina ³¹, India ³² Pakistan ³³ and Africa ³⁴ where mycotoxins occurrence is much higher. Although we couldn’t find any AFB1 data for the UK, in Scotland peanut butter and rice were found to be contaminated above the legal limits³⁵. In light of this, it is reasonable to assume higher AFB1 exposures in UK populations in comparison to France or Spain, where total imports are significantly lower.

It is important to emphasise that Germany, France, and Netherlands made up 60% of mean AFB1 estimates in Table. 3. These three countries have more funding, better equipment and more trained staff to perform the extremely costly and complex analysis. The same can be said for their agricultural sector, which is more geared towards good agricultural practices that reduce AFB1. Unfortunately, the same can not be said for other EU countries and this puts these populations at higher risk.

Lastly it is worth mentioning that mean AFB1 concentrations were determined using numerous analytical techniques. Most of which vary considerably in their limits of detection (LOD) and limits of quantification (LOQ) ³⁶. These two crucial parameters determine the level of accuracy and precision obtained in a given analyte. Furthermore, different methods can be used to calculate detection limits. These include visual evaluation, signal-to-noise, standard deviation of blank and curve calibration ³⁷. Together, these variables would have introduced greater levels of uncertainties in our AFB1 exposure estimates.

Special Consideration for Children

The existing legal maximum limits are set for food in adults only, with no consideration for children who usually consume the same foods ³⁸.

Children always account for the higher exposures because of the strong influence of body weight on exposure calculation with respect to food intake. For example, the average weight for a 10 year old child is ~25kg, making their AFB1 PMTDI 25ng per day. Therefore just one tablespoon of peanut butter (~ 30ng AFB1) would exceed their daily limit.

According to our estimates, children following vegan, vegan keto or WFBP diets could be exceeding AFB1 PMTDI by a magnitude of 4 or even more!

Reducing Aflatoxin B1

With questionable safety limits and chronic daily exposures, reducing dietary aflatoxin to as low as possible is extremely important. Luckily, there are several quick and easy methods available (Table. 4).

It is very likely that using a combination of soaking, washing and pressure cooking will result in total AFB1 elimination in all seed types (grains, legumes, beans, nuts, seeds etc..).

Unfortunately it is not possible to achieve the same flavours and textures when pressure cooking nuts and seeds. We recommend sourcing these ingredients from businesses and farmers that implement good agricultural practices. Regular peanut butter can be replaced with jungle peanut butter which is naturally free from aflatoxins. Alternatively, look for peanut products made from soaked and washed peanuts.

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