Fact sheet:

Aflatoxin B1

Version: 09/01/2015

GMP+ International B.V.

All rights reserved. The information in this publication may

be consulted on the screen, downloaded and printed as long
as this is done for your own, non-commercial use. For other
desired uses, prior written permission should be obtained
from the GMP+ International B.V. Address

DISCLAIMER: In spite of all the care and attention given to

the content of this fact sheet, it is possible that the
information is incomplete and/or not fully up-to-date. GMP+
International B.V. is not liable for any inaccuracies in this
fact sheet. You may therefore not derive any rights from this info@gmpplus.org
text. www.gmpplus.org
 Content

General Summary

Summary of GMP+ products standards for the animal feed sector

More Facts
1. Nature, history and prevalence of aflatoxin b1
2. Transmission to the environment, plants, animals and humans
3. Diagnose of poisoning
4. Potential hazards and adverse effects
5. Severity of the hazard
6. Standards
7. Analysis methods
8. Control measures
9. References
10. Websites

APPENDIX / APPENDICES

Fact sheet: Aflatoxin B1 2 / 25

Version: 09/01/2015 GMP+ International B.V.
 General Summary

Name: Aflatoxin B1

code: C01

Description: A mycotoxin (a toxic metabolic product from moulds) which is produced by the
moulds Aspergillus falvus and A. parasiticus.
Aflatoxins are very toxic and carcinogenic for the liver. Aflatoxins are mostly found on
agricultural products from tropical and subtropical areas. Weather conditions during
growing and especially during flowering have a major influence on the toxin level.
Type: chemical

Severity: high

Control measures: - (Locked) closed storage

- (Locked) closed transport
- After gathering direct transport to the client, with the advice To be fed direct or
silage / acidification'.
- Analyse intermediate product(s)
- Avoid accumulations
- Correct storage duration and temperature
- Cultivation measures
- Decrease chance of infection
- Dry or aerate products
- Drying
- Entry and reception check
- Establish purchasing requirements
- EWS
- Finished product analysis
- Gathering in a clean and dry location
- Harvesting conditions
- Maximum / low AW
- Maximum temperature
- Moisture analysis
- Preservation
- Prevention of overheating: max AW, storage duration and temperature
- Purchase requirements + inspection in connection with concentration effect.
- Quick cooling after pelletizing
- Received product/raw material analysis
- Start culture addition
- Sterilization time and temperature
- Supplier assessment
- Supplier contract
- Training farmers on Good Agricultural Practices
- Variety choice

The control measures specified in this fact sheet are all control measures which can be used depending on the
product and/or process step.

Fact sheet: Aflatoxin B1 3 / 25

Version: 09/01/2015 GMP+ International B.V.
 SUMMARY OF GMP+ SPECIFIC FEED SAFETY LIMITS FOR THE ANIMAL FEED SECTOR
Contaminant Product Action limit(1) Rejection limit(1) Source Supplementary Analysis method7
requirements
Chemical: Mycotoxin

C1 Aflatoxin B1 Feed materials intended for (direct) delivery to - 0.005 mg/kg GMP+ OZM Part 2;
dairy farmers OSP1a to 1e

Feed materials - 0.02 mg/kg Commission

Regulation (EU) No
574/2011 amending
Complementary and complete feed with the - 0.01 mg/kg Annex I to Directive
exception of: 2002/32/EC

- compound feed for dairy cattle and calves, dairy - 0.005 mg/kg
sheep and lambs, dairy goats and kids, piglets and
young poultry animals.

- compound feed for cattle (except dairy cattle and - 0,02 mg/kg
calves), sheep (except dairy sheep and lambs),
goats (except dairy goats and kids), pigs (except
piglets) and poultry (except young animals).

[1 ] Action limit: A feasible limit agreed in consultation with the sector, supplier or customer. If this limit is exceeded then an investigation into the cause should be undertaken and corrective measures should be
taken to remove or control that cause. Maximum levels in mg/kg (ppm) of the feed materials or compound feeds, derived to a moisture content of 12% unless mentioned differently.
Rejection limit: A feasible limit agreed in consultation with the sector, supplier or customer. If this limit is exceeded then the product is not suitable for use as feed material or animal feed. Maximum levels in mg/kg
(ppm) of the feed materials or compound feeds, derived to a moisture content of 12% unless mentioned differently.
[7] The research methods (OZM) can be found via the PDV website (www.pdv.nl ; quality; research methods)

Fact sheet: Aflatoxin B1 4 / 25

Version: 09/01/2015 GMP+ International B.V.
 More facts

Chemical name
(6aR-cis)-2,3,6a,9a-Tetrahydro-4-methoxycyclopenta(c)furo[3,2:4,5]furo[2,3-h][1]-
benzopyran-1,11-dione.

CAS-number
1162-65-8.

Synonyms
2,3,6a,9a-tetrahydro-4-methoxycyclopenta[c]furo[3,2:4,5]furo[2,3-h][l]benzopyran-1,11-
dione; 6-methoxydifurocoumarone; Afla B1; AFB1.

In this fact sheet will be referred to aflatoxin B1 as AFB1.

Additional information about moulds can be found in the fact sheet Moulds of GMP+
International.

1. Nature, history and prevalence aflatoxin B1

Aflatoxins are a group of mycotoxins (difuranocoumarins) produced by the Aspergillus
fungus. Although 20 compounds have been isolated, the term aflatoxins usually refers to 4
compounds produced by A. flavus and A. parasiticus, named B1, B2, G1 and G2. The 4
substances are distinguished on the basis of their fluorescent colour, B standing for blue and
G for green (WHO3, 1979). A. flavus and A. parasiticus are especially found in areas with hot,
humid climates. A. flavus is favouring the aerial parts of plants (leaves, flowers) and
produces B aflatoxins. A. parasiticus produces both B and G aflatoxins, is more adapted to a
soil environment and has more limited distribution (EFSA1, 2007). A. flavus may also produce
cyclopiazonic acid (CPA), which is toxic to a variety of animals and has been implicated in
human poisoning. CPA and aflatoxins commonly occur together in contaminated agricultural
commodities (Horn and Dorner, 1999). Speijers and Speijers (2004) indicate that AFB1 can
also be found together with mycotoxins ochratoxin and mycotoxins produced by Fusarium
fungi.

Aflatoxins are found in food and feed1 of vegetal origin as a result of fungal contamination
both pre- and post- harvest, with the rate and degree of contamination dependent on
temperature, humidity, soil and storage conditions. Although exposure to aflatoxins is
generally considered to occur mainly from imported materials, contamination of European
agricultural products cannot be excluded (EFSA2, 2004).

Among the naturally occurring aflatoxins (B1, B2, G1 and G2), AFB1 is the most important
compound with respect to both, prevalence and toxicity for humans and animals (EFSA2,
2004). Cows fed rations containing AFB1 and B2 excrete metabolites in the milk called
aflatoxin M1 and M2 (M stands for milk) (WHO3, 1979; EFSA2, 2004). Hereafter called AFM1
and AFM2.

AFB1 has a low solubility in water (Elmholt, 2008). In moderately polar solvents e.g.,
chloroform and methanol, aflatoxins are freely soluble. As pure substances, the aflatoxins
are very stable at high temperatures. However, they are relatively unstable, when exposed to
light (WHO3, 1979).
The molecular formula of AFB1 is C17H12O6. For the chemical structure of AFB1, see figure 1.

1
For the definition of feed, see GMP+ standard A2

Fact sheet: Aflatoxin B1 5 / 25

Version: 09/01/2015 GMP+ International B.V.
 Figure 1. Chemical structure AFB1

2. Transmission and likelihood of occurrence

Environment
As stated by the EFSA1 (2007) aflatoxins are a result of fungal contamination both pre- and
post-harvest, with the rate and degree of contamination dependent on temperature, humidity,
soil and storage conditions. Harvest conditions are also of importance. Several of these
environmental pre-harvest, harvest and post-harvest conditions are addressed below.
The agricultural soil serves as a reservoir for populations of A. flavus (Horn and Dorner,
1999) and A. parasiticus (Starr and Selim, 2008). Uncultivated soils contain low amounts
of A. flavus. Cultivated soil is indicated as the primary source of A. flavus and A.
parasiticus in groundnuts (IARC2, 2002). Crops grown in different soil types may have
significantly different levels of aflatoxin contamination. For example, ground nuts grown in
light sandy soils support rapid growth of the fungi, particularly under dry conditions.
Heavier soils result in less contamination of groundnuts due to their high water holding
capacity, which helps the plant to prevent drought stress (USDA, 2009).
In an experiment of Angle and Wagner (1980) the decomposition of aflatoxin in soil was
studied. AFB1 was rapidly reduced to aflatoxin B2 when added to the soil.
Studies concerning groundwater AFB1 contamination leached from soil are inconsistent.
The results of groundwater leaching studies are contradictory. Goldberg and Angle
(1985) state that AFB1 contamination of groundwater is not expected unless the soils are
extremely sandy or shallow. These authors demonstrated that AFB1 retains in the upper
20 cm. No AFB1 was found in the leachate from the soils. On the other hand Starr and
Selim (2008) state that groundwater may be contaminated via leaching, and surface
waters during rains resulting in runoff.
Regional differences in aflatoxin contamination of crops may be attributable to climatic
conditions (Horn and Dorner, 1999). Tropical conditions (high temperature and high
humidity) favour the development of aflatoxins, and contamination is usually higher in
crops grown in tropical climates than in temperate climates (USDA, 2009).
A. flavus is reported to grow within the temperature range 10-43C. The optimal growth
rate occurs at a little above 30C. The aflatoxins are produced by A. flavus over the
temperature range 15 - 37C, at least. It is not possible to specify an optimum
temperature for the production of the toxins, although production between 20-30C is
reported to be significantly greater than at higher and lower temperatures. The effect of
temperature on the behaviour of A. parasiticus is similar to that described for A. flavus
(FAO1, 2001).
The optimal water activity for growth of A. flavus and A. parasiticus is high (about 0.99).
The minimum water activity for growth has not been defined precisely. In general,
production of toxins appears to be favoured by high water activity. Crop moisture
content of more than 12% - 14% favours aflatoxin production during storage (FAO1,
2001, US-ISU, 2012) depending on the storage time. On the other hand drought stress
accompanied by elevated temperatures during seed development promotes A. flavus
invasion and subsequent aflatoxin contamination of crops (Horn and Dorner, 1999).
Drought stress is a major factor to contribute to pre-harvest aflatoxin contamination of
crop (Guo et al., 2008).
High crop density increases the aflatoxin production (USDA, 2009).

Fact sheet: Aflatoxin B1 6 / 25

Version: 09/01/2015 GMP+ International B.V.
 Weed competition: increases aflatoxin production (USDA, 2009).
Damage of crop may occur during cultivation, harvest, transport, drying or storage.
During cultivation insect infestations can increase the occurrence of Aspergillus and
levels of aflatoxins, especially during pollination in drought-stressed corn. Insect damage
to the corn ear allows easy access of Aspergillus, which subsequently infects the kernels.
Insects also transport Aspergillus spores to the silks and kernels thereby facilitating
aflatoxin contamination (USDA, 2009). Broken kernels are also subject to fungal
infection. This is caused by e.g. harvesting machines and other machines or equipment
in contact with the crop (Bruns, 2003).
Storage conditions: temperature, humidity, presence of insects and rodents. Harvested
crops that are stored under conditions favourable for Aspergillus spp. growth and
aflatoxin production are of risk. Aflatoxins can be produced in a stored crop with a
moisture content above 12%-14% and at storage temperatures greater than 21C
(FAO1, 2001; US-ISU, 2012). Residues of previously stored / handled crops can
contaminate subsequent stored / handled crops (US-ISU, 2012).

Plants
Some crop genotypes are susceptible to Aspergillus infection and insect or microbial
infestation and increase the risk of aflatoxin contamination (USDA, 2009). Damaged crops
are also more susceptible for infestation.
Agricultural soil serves as a reservoir for populations of A. flavus and infect crops (e.g.
ground nuts) in direct contact with soil populations, whereas above-ground crops, such as
corn and cottonseed, may be infected with spores from soil through dispersal by wind or
insects (Horn and Dorner, 1999). A small percentage of the AFB1 in soil is taken up by plants
(Mertz et al., 1981). Crop debris may be infected with Aspergillus spp. and serve as a
reservoir (Horn et al., 1995). For example high numbers build up because some peanuts are
not harvested but remain in the ground and act as a nutrient source (IARC2, 2002).

The food and feed most likely to be contaminated with aflatoxins are shown in table 1.
Products derived from processing thereof are also likely to be contaminated with aflatoxins.
Some examples of derived products are given in table 1, however these are not exhaustive.
Other derived products can also be of risk.
2 1 1
Table 1. Aflatoxin (B1, B2, G1, G2) risk food and feed (EFSA , 2004; EFSA , 2007; GMP+ , 2013).
*
Feed / food
Cocoa bean
Coconut, including:
Coconut expeller
Coconut extracted
Cotton seed
Cotton seed expeller
Cotton seed extracted
Fruits, dried
Groundnut, including:
Ground nut expeller
Groundnut extracted
Maize, including:
Germ
Gluten
Silage
Palm kernel, including:
Palm kernel expeller
Palm kernel extracted
Rice, including:
Rice bran
Safflower seed, including:
Safflower seed expeller
Safflower seed extracted

Fact sheet: Aflatoxin B1 7 / 25

Version: 09/01/2015 GMP+ International B.V.
 *
Feed / food
Spices
Sunflower seed, including:
Sunflower seed expeller
Tree nut, including:
Almonds
Cashew nuts
Hazelnuts
Vegetable oils, crude
*
Products derived from these products are also of risk

Petterson (2004) addresses the susceptibility of oilseeds and their by-products related to
aflatoxin. The oilseeds produced in tropical and subtropical areas are often infected by A.
flavus or A. parasiticus during both plant growth and storage and are consequently
contaminated with aflatoxins. The high water activity in oilseeds in relation to moisture
content may also promote growth of fungi and the production of aflatoxins. Oilseed expeller
and extracted meals are often transported by ship from the production place to the
consumption country and, during transport they may easily become moist, leading to
continued fungal growth and toxin production.

With respect to feed originating from Europe, only few data are available, as aflatoxin
formation was previously considered to occur mainly in geographic regions with a tropical or
subtropical climate. However, recently some reports have challenged this general
assumption. In 2013 maize intended for animal feed from Bulgaria, Hungary, Romania and
Serbia was contaminated with AFB1 causing a rise of AFM1 levels in milk in several EU
member states. The reported AFB1 levels in maize were up to 204 ppb (DG SANCO, 2013).
In 2003, Italy reported an increase in the number of milk samples exceeding the statutory
limit of AFM1, which were linked to a high contamination of locally grown maize that was
used as animal feed. Thus, although exposure to aflatoxins is generally considered to occur
mainly from imported materials, contamination of European agricultural products cannot be
entirely excluded (EFSA2, 2004).

Processing influences the AFB1 content. Wet milling reduces the concentration of aflatoxin in
maize starch, destined for food, to 1% of the levels found in the raw grain. Similarly dry
milling reduces aflatoxin in food products (grits, low-fat meal and low-fat flour) to 6-10% of
the original concentrations (WHO1, 1998). Mind that this reduction of AFB1 in food may
consequently increase the AFB1 level in feed.
In groundnuts roasting reduces aflatoxin levels by 50-80% (WHO1, 1998). The AFB1 content
in wheat is also affected by heating. Hwang and Lee (2006) reported that the level of AFB1 in
dried wheat was decreased to 50% and 90% by heating at 150 and 200C, respectively.
When 10% water was intentionally added to the wheat, the reduction was even higher.
Under unfavourable circumstances during ensiling high temperature can develop, followed
by mould growth and subsequent AFB1 production (EFSA2, 2004). Cavallarin et al. (2011)
found that aflatoxins accumulated in whole crop maize silages as a result of aerobic
exposure.
In the removal of oil from oilseeds, most of the aflatoxins are found in the oilseed meal. Small
amounts remaining in the crude vegetable oil are mainly taken out in the soap stock, the by-
product from the alkali-refining step. The remaining traces of aflatoxins are removed in the
bleaching refining steps to give aflatoxin-free refined oil (Parker and Melnick, 1966). However
residues may be present in unrefined oil (Idris et al., 2010).

AFB1 data of feed of vegetal origin can be found in the GMP+ Monitoring database of GMP+
International.

Fact sheet: Aflatoxin B1 8 / 25

Version: 09/01/2015 GMP+ International B.V.
 Animals
Exposure
AFB1 exposure occurs via feed of vegetal origin. Risk products are mentioned above in the
paragraph Plants. No data were found concerning oral AFB1 and AFM1 exposure from
feed of animal origin, e.g. via dairy products.

AFB1 data in feed can be found in the GMP+ Monitoring database of GMP+ International.

Absorption
AFB1: Rodent studies have shown that under normal conditions 50% of the orally
administered dose of AFB1 is quickly absorbed from the duodenal region of the small
intestine (Myknnen et al., 2005).
In ruminants, AFB1 is absorbed rapidly through the rumen wall (Moschini et al., 2007) and a
considerable part of the ingested AFB1 is degraded in the rumen (EFSA2, 2004).
AFM1: No data were found.

Distribution
AFB1: In ruminants the absorbed AFB1 is distributed to several tissues, e.g. the liver, kidney,
spleen, mammary gland and bile (Hayes et al., 1977). In poultry orally exposed to AFB1
residues were found in liver and muscles (Hussain et al., 2010). Oral AFB1 exposure in pigs
resulted in AFB1 residues in liver, kidney and muscle (Beaver et al., 1990). Several AFB1
metabolites are also detected in the above-mentioned tissues, depending on the animal
species (Hayes et al., 1977; Beaver et al., 1990; Hussain et al., 2010).
Placental transfer is suggested by several authors (Kihara et al., 2000; Wangikar et al.,
2005).
AFM1: No data were found.

Metabolism
AFB1: In ruminants the absorbed fraction of AFB1 is extensively metabolised in the liver,
resulting predominantly in AFM1, which enters the systemic circulation or it is conjugated to
glucoronic acid, and subsequently excreted via bile (EFSA2, 2004).
AFM1: No data were found.

Excretion
AFB1: Lthy et al. (1980) studied the metabolism and tissue distribution of AFB1 in pigs. The
major excretory route was found to be the faeces (accounting for 51 and 65% of the dose in
the two pigs). Less than 20% of the dose was excreted in the urine.
AFM1: In ruminants AFM1 can be excreted via the kidneys or appears in milk. It can also be
conjugated to glucuronic acid, and subsequently excreted via bile (EFSA2, 2004).

Humans via animal products

AFB1: There is AFB1 carry-over to edible animal tissues / products. Distribution to kidney,
liver and muscles has been established (Hayes et al., 1977; Stubblefield et al., 1981; Beaver
et al., 1990; Hussain et al., 2010). However no AFB1 data in edible products of animal origin,
besides eggs with a carry-over of 0.02% (Oliveira et al., 2000), were found. No products of
animal origin are therefore indicated as AFB1 risk products.

AFM1: Milk is contaminated with AFM1, following exposure of lactating animals to AFB1
present in feed. This carry-over rate varies in individual animals, from day to day and from
one milking to the next as it is influenced by various factors, including the feeding regime,
health status and individual biotransformation capacity, and finally by the actual milk
production. For high yielding dairy cows with a production of up to 40 litres of milk per day,
carry-over percentages as high as 6.2 % have been reported (EFSA2, 2004). Petterson

Fact sheet: Aflatoxin B1 9 / 25

Version: 09/01/2015 GMP+ International B.V.
 (2004) reviewed AFB1 (feed) -AFM1 (milk) transfer data and concluded that the mean
transfer of AFB1 in feed to AFM1 in milk in high milk producing (>25kg/day) is 2.66 1.24%.
Milk is processed into various dairy products. The processing techniques used affect the
AFM1 content in these products, however experimental results of various authors are
contradictory.
The results of studies on heat processing of dairy products on the amount of AFM1 are
ambiguous but most of the studies indicate that treatments like pasteurization and
sterilization do not cause an appreciable change in the amount of AFM1 in these products
(Prandini et al., 2009).
Evaporated, concentrated or dried milks are results of a partial or complete removal of water
from milk, with or without heating, that leads to a concentration of milk solids and
contaminants such as AFM1; this may make the toxin more susceptible to oxygen, light or
other destabilizing factors. Large losses of AFM1 were reported in some studies, whereas in
other studies milk concentration did not affect the AFM1 content (Prandini et al., 2009).
AFM1 is mainly soluble in the aqueous phase of milk or adsorbed to casein particles; data of
several studies show that a small proportion of AFM1 in milk is carried-over to cream, and yet
a smaller proportion to butter. The remainder of AFM1 in milk, however, remains in skim milk
and buttermilk (Prandini et al., 2009).
AFM1 seems to be predominantly associated with casein, so that cheese curd contains a
higher concentration than whey. Association of AFM1 with casein can be expressed as an
enrichment factor (EF) for AFM1 during cheese-making. Studies showed that the
concentration of AFM1 is about 3 fold higher in many soft cheeses and about 5 fold higher in
hard cheeses than in milk (Prandini et al., 2009).
In summary milk and dairy products should be considered as AFM1 risk product.

Humans
Exposure
AFB1: No human AFB1 exposure data via products of products of animal origin were found.
The most important source of human AFB1 exposure is via products of vegetal origin.
Though a wide range of foods may be contaminated with aflatoxins (B1, B2, G1, G2), they
have been most commonly associated with tree nuts, groundnuts, figs and other dried fruits,
spices, crude vegetable oils, cocoa beans, maize, rice, cottonseed and copra.
The EU in the Scientific Cooperation Assessment Project (SCOOP) on aflatoxins indicated
an intake range of 0.03 to 1.3 g/kg bodyweight per day for AFB1. Population groups with
high nut consumption are exposed to higher levels of aflatoxins (EFSA1, 2007).
Data from Australia indicated an average estimated intake of 0.15 g/kg bodyweight per day
of aflatoxins. A series of Chinese studies reported intakes ranging from 0 to 91 g/kg
bodyweight per day of AFB1.
AFM1: Exposure occurs via diary products, following exposure of lactating animals to AFB1
in feed (EFSA2, 2004).

Unfortunately data reported in literature on the toxicokinetics (absorption, distribution,

metabolism and excretion) of AFM1 are scarce (Caloni1 et al., 2011) and are only discussed
briefly below.

Absorption
AFB1: After oral uptake AFB1 is efficiently absorbed and metabolized prior to excretion by
faecal and urinary routes (Myknnen et al, 2005).
AFM1: absorption, however poorly, of AFM1 has been detected using a human in vitro model
(Caloni2 et al., 2006; Caloni1, 2011).

Distribution
AFB1 is concentrated in the liver and to a lesser extent in kidney, and it is also found in the
blood (in the intestine area) as free AFB1 or its water-soluble metabolites (Myknnen et al,
2005).

Fact sheet: Aflatoxin B1 10 / 25

Version: 09/01/2015 GMP+ International B.V.
 AFM1: no data were found.
Aflatoxins cross the human placenta (IARC2, 2002).

Metabolism
AFB1: The liver is the major site of aflatoxin metabolism. In the liver enzymes, have been
shown to be responsible for the metabolism of the absorbed aflatoxins. These enzymes
convert AFB1 to its carcinogenic form and can also oxidize AFB1 to various other
derivatives, including AFM1 (Myknnen et al, 2005).
AFM1: no data were found.

Excretion
AFB1: Absorbed AFB1 and its metabolites are excreted in urine, while elimination to faeces
is a route for both the unabsorbed AFB1 and biliary excretion of metabolites formed from the
absorbed toxin (Myknnen et al, 2005).
AFM1: AFM1 is excreted in human breast milk (Sadeghi et al., 2009) and urine (IARC2,
2002).

3. Diagnose of poisoning

Animals
Since milk contaminated with the AFM1, following exposure of lactating animals to AFB1
present in feed, it is expected that milk can be used as a biomarker for AFB1 exposure.
Possibly other biomarkers can be used that are present in urine, faeces or bile.
Disease history, necropsy findings, and microscopic examination of the liver should indicate
the nature of the liver toxin, but liver changes are somewhat similar in Senecio poisoning.
The liver shows proliferation and fibrosis of the bile ductules (Kahn, 2005).
Feed can be analysed for AFB1 and AFM1.

Humans
AFB1 metabolites can be useful biomarkers of human exposure to aflatoxins and AFM1,
aflatoxin Q1 and aflatoxin P1 be excreted in urine and bile and have all been detected in
human urine samples. Also the aflatoxin-albumin adduct in serum and plasma together with
urinary aflatoxin metabolites and adducts have been valuable biomarkers of aflatoxin
exposure in epidemiological studies (EFSA1, 2007). AFM1 is excreted in human breast milk,
which can be used as a biomarker for lactating mothers exposure to AFB1 (Sadeghi et al.,
2009).
Food can be analysed for AFB1 and AFM1.

4. Potential adverse effects

AFM1 is less toxic than AFB1.

Environment
No data were studied.

Animals
AFB1: Aflatoxicosis affects young animals: growing poultry (especially ducklings and
turkey poults), young pigs, pregnant sows and calves. Adult cattle, sheep and goats are
relatively resistant to the acute form of the diseasebut are susceptible if toxic feed is fed over
long periods (Kahn, 2005). Bovine species are generally less sensitive compared to non-
ruminants because aflatoxins are partly degraded by the fore-stomach flora, however dairy
cattle is of specific risk for its transmission of AFB1 into AFM1in milk (EFSA2, 2004).

Fact sheet: Aflatoxin B1 11 / 25

Version: 09/01/2015 GMP+ International B.V.
 The liver is the primary target in different animal species (Wijnands and Van Leusen, 2000).
Experimental and field studies indicated that long term exposure to relatively low
concentrations of aflatoxins may result in hepatic fibrosis and liver cell tumours in birds, pigs,
and sheep (EFSA2, 2004). The International Agency for Research on Cancer (IARC1, 1993)
states that there is sufficient evidence in experimental animals for the carcinogenicity of
naturally occurring mixtures of aflatoxins and of AFB1. Mutagenicity is reported (Wild and
Turner, 2002; IARC3, 2012), however specific data concerning tested animal species and
experimental designs were not found.
AFB1 is also immunosuppressive in animals. Exposure to aflatoxin results in increased
susceptibility to bacterial and parasitic infections (IARC2, 2002). This is confirmed by the
Food and Agriculture Organization of the United Nations (FAO2, 1994), stating that chronic
exposure with low levels of aflatoxin have been associated with an increased susceptibility to
disease in poultry, pigs and cattle. Vaccine failures have also been reported. Reduced weight
gain has also been reported, for example, in cattle, pigs and poultry (FAO2, 1994).
The results of a study by Wangikar et al. (2005) indicated that AFB1 was found to be toxic to
rabbit embryos when given by oral route during gestation.

AFM1: No data were found concerning susceptible animal species.

The IARC3 (2012) states that there is sufficient evidence for the carcinogenicity in
experimental animals of naturally occurring mixtures of aflatoxins and of the individual AFM1
Mind that aflatoxins can interact with other mycotoxins, also present in feed. Interaction with
other mycotoxins can lead to synergistic, additive and antagonistic effects (Speijers and
Speijers, 2004).

In Appendix I the potential adverse effects of AFB1 and AFM1 in animals are shown. For
toxicity data, see Appendix II.

Humans
AFB1: The IARC1 (1993) states that there is sufficient evidence in humans for the
carcinogenicity of AFB1.
In young children aflatoxin exposure has been associated with growth impairment (IARC2,
2002), underweight (USDA, 2009) and impairment of cellular immunity that could decrease
host resistance to infections (EFSA1, 2007). They become more susceptible to infectious
diseases such as malaria, diarrhoea, and respiratory infections in childhood and later in life.
Several authors report mutagenicity of AFB1 (Wild and Turner, 2002; IARC3, 2012).
Gastrointestinal bleedings have been observed in several cases of aflatoxicosis (WHO4,
1999).

AFM1: Since AFM1 is excreted in human and animal milk, infants and young children are
exposed to high levels of AFM1 and are therefore more susceptible for toxic AFM1 effects.
The IARC has classified AFM1 as a possible human carcinogen (group 2b) (IARC1, 1993).

In Appendix I the potential adverse effects of AFB1 and AFM1 in humans are shown. For
toxicity data, see Appendix II.

Fact sheet: Aflatoxin B1 12 / 25

Version: 09/01/2015 GMP+ International B.V.
 5. Severity of the potential adverse effects
The severity of contamination of food and / or feed with AFB1 and AFM1 is based on the
worst case scenario, see table 2 and 3 respectively.

The severity of AFB1 toxicosis in animals is classified as high because:

The IARC1 (1993) states that there is sufficient evidence in experimental animals for the
carcinogenicity of naturally occurring mixtures of aflatoxins and aflatoxins B1, G1 and M1;
Teratogenic effects (toxic to embryos) are observed (IARC2, 2002);
Mutagenicity has been reported by several authors (Wild and Turner, 2002; IARC3,
2012).

The severity of AFB1 toxicosis in humans is classified as high because:

The IARC1 (1993) states that there is sufficient evidence in humans for the
carcinogenicity of AFB1;
Impairment of cellular immunity has been observed (EFSA1, 2007);
Mutagenicity is reported by many authors (Wild and Turner, 2002; IARC3, 2012);

Table 2. Severity of AFB1

Severity
Low Medium High
Animals x
Humans x

The severity of AFM1 toxicosis in animals is classified as high because:

The IARC3 (2012) states that there is sufficient evidence in experimental animals for the
carcinogenicity of naturally occurring AFM1;

The severity of AFM1 toxicosis in humans is classified as high because:

The IARC has classified AFM1 as a possible human carcinogen (group 2b) (IARC1,
1993).

Table 3. Severity of AFM1

Severity
Low Medium High
Animals x
Humans x

6. Legislation and standards

AFB1: The FDA has stated AFB1 action levels in feed for different animal species (US-FDA,
2009). These maxima tend to be higher than maxima within the European Union.
Several countries have regulations, standards or guidelines for AFB1 in feed. Also several
market driven concepts within countries might have standards or guidelines for AFB1.
No other standards were studied.
AFM1: no data related to feed were found.

Fact sheet: Aflatoxin B1 13 / 25

Version: 09/01/2015 GMP+ International B.V.
 7. Method of analysis
Sampling: In GMP+ standards BA4 and BA13 requirements concerning sampling are stated
(GMP+1,2, 2013).

Analysis: For the monitoring of the presence of aflatoxins in food and feed, various validated
methods of analysis exist. In GMP+ standard BA4 requirements concerning analysis are
stated (GMP+1, 2013).
AFB1: For the determination of AFB1 these methods are based on solid phase extraction
(SPE) cleanup in combination with liquid chromatography, and on immuno affinity (IA)
cleanup in combination with liquid chromatography (EFSA2, 2004). The PDV suggests HPLC
or HPLC iodine or bromine derivatisation, depending on matrix and detection limits (PDV1,
1995; PDV2, 1996; PDV3, 1996; PDV4, 2002; PDV5, 2005).
AFM1: In dairy product the confirmation of the presence of AFM1 is done with SPE and high-
performance liquid chromatography (HPLC). Milk samples can be screened to detect the
presence of AFM1 using enzyme-linked immunosorbent assay (ELISA). ELISA is not fully
reliable due to cross-reaction interferences (Chen et al., 2005). No analysis data were found
concerning other matrices.

8. Possible control measures

Aflatoxins presence in food and feed is a result of mould contamination both pre- and post
harvest. Control measures should focus on these parts.

The HACCP system should be used to assess the risk of moulds and aflatoxins (AFB1 and
AFM1) in feed and consequently the control measures needed to control this risk. All parts in
the feed and food chain should be included, starting at cultivation or husbandry. For more
information concerning possible mould control measures in general is referred to the fact
sheet Moulds of GMP+ International. In Appendix III of the fact sheet Moulds the possible
general undesirable moulds control measures are mentioned, applicable for several part in
the feed and food chain. These general control measures can be complemented with the
specific possible mould control measures for specific parts in the feed and food chain. These
specific control measures are included in the appendices of the fact sheet Moulds.

In addition to the control measures as mentioned in the fact sheet Moulds there are specific
aflatoxin control measures. These are also related to specific activities within each part of the
feed and food chain. The specific control measures are included in the appendices:
Cultivation, harvesting, processing, transport and storage on farm, see Appendix III;
Husbandry, see Appendix IV;
Transport, see Appendix V;
Storage, see Appendix VI;
Processing of feed, see Appendix VII.

The possible control measures should focus on products with a higher risk to contain
aflatoxins (AFB1 and AFM1) are summarised in table 4. It should be kept in mind that also
products derived from the risk products mentioned are also of risk. Examples of derived
products are given in table 4, however these examples are not exhaustive. Other derived
products can also be of risk.

Fact sheet: Aflatoxin B1 14 / 25

Version: 09/01/2015 GMP+ International B.V.
 Table 4. Products with a higher risk to contain aflatoxins (AFB1 and AFM1).
Crop with a moisture content of more than 12% - 14%;
vegetal origin

Products of

Damaged crop;
in general

Crop debris;
Harvested crops that are stored under conditions favourable for Aspergillus spp. growth
and aflatoxin production: stored crop with a moisture content above 12%-14% and at
storage temperatures greater than 21C;
Residues of previously stored / handled crops.
Cocoa beans;
Specific products of vegetal origin

Coconut: expeller and extracted;

Cotton seed: expeller and extracted;
Fruits, dried;
Groundnuts: expeller and extracted;
Maize, including germ, gluten, silage;
Palm kernel, including expeller and extracted;
Rice, including bran;
Tree nuts, including almonds, cashew nuts, hazelnuts;
Safflower seed, including expeller and extracted
Spices
Sunflower seed, including expeller and extracted;
Vegetable oils, crude

Milk and other dairy products

of animal
Products

origin

Aflatoxicosis affects young animals: growing poultry (especially ducklings and turkey
poults), young pigs, pregnant sows and calves. Dairy cattle are not sensitive species as
such however the transmission of AFB1 to AFM1 in milk makes dairy cattle a critical species
related to the high level of AFM1 exposure of infants and young children via milk and dairy
products.

9. References
1. Angle and Wagner, Decomposition of aflatoxin in soil, Soil Science Society of American
Journal, Volume 40, 1980, pages 1237-1240
2. Beaver et al., Distribution of aflatoxin in tissues of growing pigs fed an aflatoxin-
contaminated diet amended with a high affinity aluminosilicate sorbent, Veterinary and
Human Toxicology, Volume 32(1), 1990, pages 16-18
3. Bruns, Controlling aflatoxin and fumonisin in maize by crop management, Journal of
Toxicology: Toxin Reviews, Volume 22(2&3), 2003, pages 153-173
4. Caloni1 et al., Transport of aflatoxin M1 in human intestinal Caco-2/TC7 cells, Frontiers in
Pharmacology, Volume 3, 2011, pages 1-11
5. Caloni2 et al., Aflatoxin M1 absorption and cytotoxicity on human intestinal in vitro model,
Toxicon, Volume 47(4), 2006, pages 409-415
6. Cavallarin et al., Aflatoxin accumulation in whole crop maize as a result of aerobic
exposure, Journal of the Science of Food and Agriculture, Volume 91(13), 2011, pages
2419-2425
7. Chen et al., Determination of aflatoxin M1 in milk and milk powder using high-flow solid-
phase extraction and liquid chromatography-tandem mass spectrometry, Journal of
Agricultural and Food Chemistry, Volume 53, 2005, pages 8474-8480
8. DG SANCO (European Health and Consumer Protection Directorate General),
Notification detail 2013.0268, RASFF Portal, 2013

Fact sheet: Aflatoxin B1 15 / 25

Version: 09/01/2015 GMP+ International B.V.
 9. EC (European Commission), Commission Recommendation of 17 August 2006 on the
prevention and reduction of Fusarium toxins in cereals and cereal products
(2006/583/EC), 2006
10. EFSA1, Opinion of the Scientific Panel on contaminants in the food chain on a request
from the Commission related to the potential increase of consumer health risk by a
possible increase of the existing maximum levels for aflatoxins in almonds, hazelnuts and
pistachios and derived products, The EFSA Journal 446, 2007
11. EFSA2, Opinion of the Scientific Panel on contaminants in the food chain on a request
from the Commission related to aflatoxin B1 as undesirable substance in animal feed, The
EFSA Journal 39, 2004
12. Elmholt, Mycotoxins in the soil environment, Secondary metabolites in soil ecology (ed.
Karlovsky), Chapter 9, 2008, pages 167293
13. FAO1 (Food and Agriculture Organization of the United Nations), Manual on the
application of the HACCP system in Mycotoxin prevention and control, 2001
14. FAO2 (Food and Agriculture Organization of the United Nations), Grain storage
techniques, FAO Agricultural Services Bulletin No. 109, 1994
15. GMP+1 International, Minimum requirements for sampling and analysis, GMP+ BA4,
2013
16. GMP+2 International, Minimum requirements for sampling, GMP+ BA13, 2013
17. GMP+3 International, GMP+ B6, Feed materials cultivation, GMP+ Feed Safety
Assurance scheme, 2013
18. GMP+5 International, GMP+ B1: Production, Trade and Services, GMP+ Feed Safety
Assurance scheme, 2013
19. GMP+6 International, GMP+ B4, Transport, GMP+ Feed Safety Assurance scheme, 2013
20. GMP+7 International, GMP+ B4.1, Road transport, GMP+ Feed Safety Assurance
scheme, 2013
21. GMP+8 International, GMP+ B4.2, Affreightment of short sea shipping and inland
waterway transport, GMP+ Feed Safety Assurance scheme, 2013
22. GMP+9 International, GMP+ B4.3, Short sea shipping and inland waterways transport,
GMP+ Feed Safety Assurance scheme, 2013
23. GMP+10 International, GMP+ B4.4, Sea transport and affreightment, GMP+ Feed Safety
Assurance scheme, 2013
24. GMP+11 International, GMP+ B4.5, Rail transport and affreightment, GMP+ Feed Safety
Assurance scheme, 2013
25. GMP+12 International, GMP+ B5, Storage & transhipment, GMP+ Feed Safety Assurance
scheme, 2013
26. GMP+13 International, GMP+ B2, Quality control of feed materials, GMP+ Feed Safety
Assurance scheme, 2013
27. Goldberg and Angle, Aflatoxin movement in soil, Journal of Environmental Quality,
Volume 14(2), 1985, pages 224-228
28. Gua et al., Drought stress and preharvest aflatoxin contamination in agriculture
commodity: genetics, genomics and proteomics, Journal of Integrative Plant Biology,
Volume 50(10), 2008, pages 1281-1291
29. Hayes et al., Bovine liver metabolism and tissue distribution of aflatoxin B1, Journal of
Agricultural and Food Chemistry, Volume 25(5), 1977, pages 1189-1193
30. Hwang and Lee, Reduction of aflatoxin B1 contamination in wheat by various cooking
treatments, Food Chemistry, Volume 98(1), 2006, pages 7-75
31. Horn and Dorner, Regional Differences in Production of Aflatoxin B1 and Cyclopiazonic
Acid by Soil Isolates of Aspergillus flavus along a Transect within the United States,
Applied and Environmental Microbiology, Volume 65(4), 1999, pages 1444-1449
32. Horn et al., Effect of corn and peanut cultivation on soil populations of Aspergillus flavus
and A. parasiticus in Southwestern Georgia, Applied and Environmental Microbiology,
Volume 61(7), 1995, pages 2472-2475

Fact sheet: Aflatoxin B1 16 / 25

Version: 09/01/2015 GMP+ International B.V.
 33. Hussain et al., Residues of alflatoxin B1 in broiler meat: Effect of age and dietary
aflatoxin B1 levels, Food and Chemical Toxicology, Volume 48(12), 2010, pages 3304-
3307
34. IARC1 (International Agency for Research on Cancer), Naturally occurring aflatoxins
(group 1) and aflatoxin M1 (group 2), Summaries & Evaluations, Volume 56, 1993, page
245
35. IARC2 (International Agency for Research on Cancer), IARC monographs on the
evaluation of carcinogenic risks to humans, Some traditional herbal medicines, some
mycotoxins, naphthalene and styrene, Volume 82, 2002
36. IARC3 (International Agency for Research on Cancer), Chemical agents and related
occupations, a review of human carcinogens, IARC Monographs, Volume 100F, 2012
37. Idris et al., Determination of aflatoxin levels in Sudanese edible oils, Food and Chemical
Toxicology, Volume 48, 2010, pages 2539-2541
38. Kahn (ed.), The Merck Veterinary Manual, Ninth edition, 2005
39. Kihara et al., Effects of prenatal aflatoxin b1 exposure on behaviors of rat offspring,
Toxicological Sciences, Volume 53(2), 2000, pages 392-399
40. Lthy et al., Metabolism and tissue distribution of [14C]-aflatoxin B1 in pigs, Food and
Cosmetics Toxicology, Volume 18(3), 1980, pages 253-256
41. Mertz et al., Absorption of aflatoxin by lettuce seedlings grown in soil adulterated with
aflatoxin B1, Journal of Agricultural and Food Chemistry, Volume 29(6), 1981, pages
1168-1170
42. Moschini et al., Mucosal absorption of aflatoxin B1 in lactating dairy cows, Italian Journal
of Animal Science, Volume 6(1), 2007, pages 324-326
43. Myknnen et al., Fecal and urinary excretion of aflatoxin B1 metabolites (AFQ1, AFM1
and AFB-N7-guanine) in young Chinese males, International. Journal of Cancer, Volume
115(6), 2005, pages 879-884
44. Oliveira et al, Aflatoxin B1 residues in eggs of laying hens fed a diet containing different
levels of the mycotoxin, Food additives and contaminants, Volume 17(6), 2000, pages
459-462
45. Parker and Melnick, Absence of aflatoxin from refined vegetable oils, The Journal of
American Oil Chemists Society, Volume 43, 1966, pages 635-638
46. PDV1, OSP-01a Aflatoxine B1: HPLC met jodiumderivatisering, 1995
47. PDV2, OSP-01b Aflatoxine B1: HPLC met broomderivatisering, 1996
48. PDV3, OSP-01c Aflatoxine B1: HPLC bepaalde diervoedergrondstoffen, 1996
49. PDV4, OSP-01e Aflatoxine B1: HPLC met broomderivatisering, 2002
50. PDV5, OSP-01d Aflatoxine B1: HPLC met broom- en jodiumderivatisering, 2005
51. Petterson, Controlling mycotoxins in animal feed. In: Mycotoxins in food, detection and
control, 2004, pages 262-304
52. Prandini et al., On the occurrence of aflatoxin M1 in milk and dairy products, Food and
Chemical Toxicology, Volume 47, 2009, pages 984-991
53. Sadeghi et al, Incidence of aflatoxin M1 in human breast milk in Tehran, Iran, Food
Control, Volume 20(1), 2009, pages 75-78
54. Speijers and Speijers, Combined toxic effects of mycotoxins, Toxicological Letters, 153,
2004, pages 91-98
55. Starr and Selim, Supercritical fluid extraction of aflatoxin B1 from soil, Journal of
Chromatography A, Volume 1209, 2008, pages 37-43
56. Stubblefield et al, Transmission and distribution of aflatoxin in contaminated beef liver
and other tissues, Journal of the American Oil Chemists Society, Volume 58(12), 1981,
pages 1015-1017
57. USDA, Aflatoxins: A Focus on Aflatoxin contamination: Updated Version, June 2009
58. US-FDA, Guidance for Industry: Action Levels for Poisonous or Deleterious Substances
in Human Food and Animal Feed: contains nonbinding recommendations, 2009
59. US-ISU (United States Iowa State University), Aflatoxins in corn, PM 1800, 2012
60. Wangikar et al., Effects of aflatoxin B1 on embryo fetal development in rabbits, Food and
Chemical Toxicology, Volume 43(4), 2005 pages 607-615

Fact sheet: Aflatoxin B1 17 / 25

Version: 09/01/2015 GMP+ International B.V.
 61. Wijnands and Van Leusden (RIVM), An overview of adverse health effects caused by
mycotoxins and bioassays for their detection, RIVM report 257852 004, 2000
62. WHO1, Safety evaluation of certain food additives and contaminants: Aflatoxins, WHO
food additives series No. 40, 1998
63. WHO2, Evaluation of certain mycotoxins in food, WHO Technical Report Series 906,
2002
64. WHO3, Mycotoxins, Environmental Health Criteria, No. 11, 1979
65. WHO4, Toxic effects of mycotoxins in humans, Bulletin of the World Health Organization,
No. 77 (9), 1999
66. Wild and Turner, The toxicology of aflatoxins as a basis for public health decisions,
Mutagenesis, Volume 17(6), 2002, pages 471-481

10. Websites
1. https://www.gmpplus.org/pagina/288/home_un.aspx
2. http://www.inchem.org/documents/jecfa/jecmono/v040je16.htm
3. http://www.inchem.org/documents/iarc/vol82/82-04.html
4. http://www.inchem.org/documents/iarc/vol56/09-afl.html
5. http://fsrio.nal.usda.gov/document_fsheet.php?product_id=226
6. http://www.efsa.europa.eu/EFSA/efsa_locale-178620753812_1178620761977.htm
7. http://www.efsa.europa.eu/EFSA/efsa_locale-178620753812_1178620763196.htm

Fact sheet: Aflatoxin B1 18 / 25

Version: 09/01/2015 GMP+ International B.V.
 APPENDIX I Potential adverse effect of aflatoxin B1 (AFB1) and aflatoxin M1 (AFM1)

2,3 2,3 2,3 2,3 3 3 3

AFB1 Death Carcinogen Mutagen Reproductive Internal injury Neurological Immunological Organs
(physical
contamination)
2,3

Animals x x x x x

Humans x x x

3 3 3 3
AFB1 Dermal and Respiratory Musculo- Cardiovascular Gastrointestinal Hematological Endocrine Body weight
3 3
ocular skeletal
Animals x

Humans x x

2,3 2,3 2,3 2,3 3 3 3

AFM1 Death Carcinogen Mutagen Reproductive Internal injury Neurological Immunological Organs
(physical
contamination)
2,3

Animals x

Humans x

3 3 3 3
AFM1 Dermal and Respiratory Musculo- Cardiovascular Gastrointestinal Hematological Endocrine Body weight
3 3
ocular skeletal
Animals

Humans

2
This potential adverse effect is classified as high severity for animals
3
This potential adverse effect is classified as high severity for humans

Fact sheet: Aflatoxin B1 19 / 25

Version: 09/01/2015 GMP+ International B.V.
 APPENDIX II Toxicity data aflatoxin B1 and M1

NOEL (mg/kg BW) ADI (mg/kg BW) LD50 (mg/kg BW)

Aflatoxin B1 30 g/kg BW/day (mice) ALARA2 (EFSA1, 2007) 5,5 (rat) (EFSA1, 2007)
(EFSA1, 2007)1
Aflatoxin M1 - - -
1
= during 4 weeks, NOEL for immune suppression
2
= ALARA = As Low As Reasonably Achievable

Fact sheet: Aflatoxin B1 20 / 25

Version: 09/01/2015 GMP+ International B.V.
 APPENDIX III Aflatoxin control measures: cultivation, harvesting, processing,
transport and storage of feed on farm.
Requirements and control measures related to feed ingredient cultivation are stated in the
GMP+ B6 standard (GMP+3, 2013). This standard should be complied with. Additional
recommendations for control measures can be found in the fact sheet Moulds. Besides the
control measures mentioned in the fact sheet Moulds there are no specific
recommendations for aflatoxins control measures.

Concerning aflatoxins monitoring programs the suggested mould monitoring programs in the
fact sheet Moulds can be used. In addition to mould monitoring programs, implementation
of aflatoxins monitoring programs is advised. As a guideline the mould monitoring programs
in the fact sheet Moulds can be used, replacing the word moulds by aflatoxins, with a
focus on AFB1 monitoring.

Fact sheet: Aflatoxin B1 21 / 25

Version: 09/01/2015 GMP+ International B.V.
 APPENDIX IV Aflatoxins control measures: husbandry.

Recommendations for control measures can be found in the fact sheet Moulds. Besides the
control measures mentioned in the fact sheet Moulds there are no specific
recommendations for aflatoxins control measures.

Concerning aflatoxins monitoring programs the suggested mould monitoring programs in the
fact sheet Moulds can be used. In addition to mould monitoring programs, implementation
of aflatoxins monitoring programs is advised. As a guideline the mould monitoring programs
in the fact sheet Moulds can be used, replacing the word moulds by aflatoxins, with a
focus on AFB1 monitoring.

Fact sheet: Aflatoxin B1 22 / 25

Version: 09/01/2015 GMP+ International B.V.
 APPENDIX V Aflatoxins control measures: transport of feed.
In GMP+ B1 standard general control measures and requirements are stated which should
be complied with (GMP+5, 2013). Requirements and control measures related to transport
are stated in various GMP+ standards (GMP+6,7,8,9,10,11, 2013). The applicable GMP+
standard depends on the means of transport. The applicable standard should be complied
with.

Recommendations for control measures can be found in the fact sheet Moulds. Besides the
control measures mentioned in the fact sheet Moulds there are no specific
recommendations for aflatoxins control measures.

Concerning aflatoxins monitoring programs the suggested mould monitoring programs in the
fact sheet Moulds can be used. In addition to mould monitoring programs, implementation
of aflatoxins monitoring programs is advised. As a guideline the mould monitoring programs
in the fact sheet Moulds can be used, replacing the word moulds by aflatoxins, with a
focus on AFB1 monitoring.

Fact sheet: Aflatoxin B1 23 / 25

Version: 09/01/2015 GMP+ International B.V.
 APPENDIX VI Aflatoxins control measures: storage of feed.
In GMP+ B1 standard general control measures and requirements are stated which should
be complied with (GMP+5, 2013). Specific requirements and control measures related to
storage are stated in the GMP+ B5 standard (GMP+12, 2013). This standard should be
complied with.

Recommendations for control measures can be found in the fact sheet Moulds. Besides the
control measures mentioned in the fact sheet Moulds there are no specific
recommendations for aflatoxins control measures.

Concerning aflatoxins monitoring programs the suggested mould monitoring programs in the
fact sheet Moulds can be used. In addition to mould monitoring programs, implementation
of aflatoxins monitoring programs is advised. As a guideline the mould monitoring programs
in the fact sheet Moulds can be used, replacing the word moulds by aflatoxins, with a
focus on AFB1 monitoring.

Fact sheet: Aflatoxin B1 24 / 25

Version: 09/01/2015 GMP+ International B.V.
 APPENDIX VII Aflatoxins control measures: processing of feed.
In GMP+ B1 and B2 standard general control measures and requirements are stated which
should be complied with (GMP+5,13, 2013). These standards should be complied with.

Recommendations for control measures can be found in the fact sheet Moulds. Besides the
control measures mentioned in the fact sheet Moulds there are no specific
recommendations for aflatoxins control measures.

Concerning aflatoxins monitoring programs the suggested mould monitoring programs in the
fact sheet Moulds can be used. In addition to mould monitoring programs, implementation
of aflatoxins monitoring programs is advised. As a guideline the mould monitoring programs
in the fact sheet Moulds can be used, replacing the word moulds by aflatoxins, with a
focus on AFB1 monitoring.

Fact sheet: Aflatoxin B1 25 / 25

Version: 09/01/2015 GMP+ International B.V.