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Fact sheet:
Moulds and yeasts
Version: 09/01/2015
GMP+ International B.V.
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Content
General Summary
Summary of GMP+ products standards for the animal feed sector
More Facts
1. Nature, history and prevalence of moulds and yeasts
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: Moulds and yeasts 2 / 34

Version: 09/01/2015 GMP+ International B.V.
General Summary
Name: Moulds and yeasts
code: M05
Description: Moulds and fungal spores always occur on the feed. Moulds can be harmful. They
can spoil feed materials, be pathogenic and produce toxic mycotoxins such as
aflatoxin (harmful metabolic products).
Type: microbiological
Severity: high
Control measures: - (Locked) closed storage

- (Locked) closed transport
- Aerobic environment (Brewers' grain)
- After gathering direct transport to the client, with the advice To be fed direct or
silage / acidification'.
- Air filtration
- Analyse intermediate product(s)
- Bark peeling (or entry check)
- Check of odour and appearance
- Chemical treatment
- CIP (cleaning in place)
- Clean cheese
- Climate/temperature control
- Closed production system
- Correct storage duration and temperature
- Covering hay bales
- Covering production area.
- Dedicated storage/transport
- Destruction of affected product
- Dry or aerate products
- Dry plant/dry storage conditions
- Drying
- Entry and reception check
- Establish purchasing requirements
- Extraction with ethanol/nitric acid
- Finished product analysis
- Gathering in a clean and dry location
- Grinding line: extra attention to cleaning
- Heating
- High salt content
- High temperature and high pressure
- Low moisture percentage
- Maximum / low AW
- Maximum Aw liquid-rich end products
- Maximum pH
- Maximum pH liquid-rich end products
- Maximum temperature
- Packaged end products
- Packaged raw materials
- Preservation
- Prevention accumulation of organic material
- Prevention of overheating: max AW, storage duration and temperature
- Purity strains control
- Quick cooling after pelletizing
- Quick processing of the product
- Received product/raw material analysis
- Rotation of crops
- Stainless steel transport/installation
- Sterilization time and temperature
- Supplier contract
- Training farmers on Good Agricultural Practices
- Use new packaging material
- Use tapwater
- Water management
The control measures specified in this fact sheet are all control measures which can be used depending on the
product and/or process step.

SUMMARY OF GMP+ SPECIFIC FEED SAFETY LIMITS FOR THE ANIMAL FEED SECTOR
Contaminant Product Action limit(1) Rejection limit(1) Source Supplementary Suggestion for
requirements analysis method
Microbiological: Microbiological contamination
6
M5a Moulds Feed materials 10 CFU/g In the TNO report Norm Enumeration NEN-
for fungal load in animal ISO 21527-1:2008
feed (D4.16) you can
read the foundation of the Enumeration NEN-
new standards and the ISO 21527-2:2008
suggestions for analysis (if aw-value 0.95)
methods.
6
M5b Yeast Feed materials 12% moisture content or 10 CFU/g Enumeration NEN-
aw-value 0.95 ISO 21527-1:2008
Feed materials 12% moisture content or aw- -

value 0.95
[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.
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.

More facts
Chemical name
Not applicable.
CAS-number
Not applicable.
Synonyms
Not applicable.
This fact sheet focuses on mould as undesirable substance.
1. Nature, history and prevalence of moulds

Moulds are part of a large microorganism group called fungi and are present in every part in
the world. The gross appearance of a mould growing on a feed is often enough to indicate its
genus. Macroscopically the mould consists of a mass of branching, intertwined filaments
called hyphae (singular hypha), and the whole mass of these hyphae is known as a
mycelium. Hyphae may be classed as vegetative or fertile based on their biological function.
The reproductive parts or structures of moulds are the spores, which are mainly asexual.
Such spores are produced in large numbers and are readily spread by air. Spores that settle
on favourable substrates can initiate a new phase of growth and develop into a new
mycelium. A few kinds of moulds produce sclerotia (singular sclerotium) which are tightly
packed masses of hyphae, often thick-walled, within the mycelium. These sclerotia are
considerably more resistant to heat and other adverse conditions than the rest of the
mycelium (Lasztity, 2009).
Moulds are not necessarily hazardous to humans and animals. They can be used
intentionally in food processing e.g. in cheese, citric acid processing. However moulds are
also related to spoilages of feed and food. A picture of mouldy grain is shown in figure 1.
Figure 1. Mouldy grain (source: ukmalt.com)
Moulds can grow pre-harvest or postharvest, during storage, processing, or feeding (Whitlow
and Hagler1, 2007). Mould growth is related to damaged crop (e.g. damaged kernels) (FAO1,
2007), weather extremes (causing plant stress or excess hydration of stored feed), to
inadequate storage practices, to low feedstuff quality, and to faulty feeding conditions. In
general, environmental conditions heat, water, and insect damage cause plant stress and
predispose plants in the field to mycotoxin contamination. Because feed can be
contaminated pre-harvest, control of additional mould growth is dependent on storage

management. After harvest, temperature, moisture content, and insect activity are the major
factors influencing mould contamination of feed and food (Coulombe, 1993).
Detrimental effects of moulds on feed are:

a) Mycotoxin production;
b) Heating and moisture increase;
c) Mustiness (staleness);
d) Loss of nutritional value
a) Mycotoxin and MVOCs production

Mycotoxins can be formed on crops in the field, during harvest, or during storage,
processing or feed (Whitlow and Hagler1, 2007). Fusarium and Claviceps are so-called
field moulds and produce mycotoxins in the field. Penicillium and Aspergillus are
storage moulds and produce mycotoxins mainly during storage (PDV1, 2003). Examples
of mycotoxins of which fact sheets are available at GMP+ International are aflatoxin B1,
deoxynivalenol, fumonisin B1, ochratoxin A and zearalenone. Examples of moulds
commonly reported in feed or food and some of the mycotoxins produced are given in
table 1. These mycotoxins are not addressed to in this fact sheet. For more information is
referred to the specific mycotoxins fact sheets available at GMP+ International.
Table 1. Fungi commonly reported in feed or food.

Fungus Mycotoxin
Aspergillus Aflatoxin
Citrinin
Fusarium Deoxynivalenol
Fumonisin
Gliotoxin
T-2 toxin
Zearalenone
Penicillium Citrinin
Ochratoxin
Patulin
PR-toxin
Roquefortine
Moulds also produce microbial volatile organic compounds (MVOCs). More than 200
compounds have been regarded as MVOCs. The compounds also have other
environmental sources than mould metabolism. Thus, compounds originating solely from
mould metabolism hardly exist (Korpi et al., 2007) and are therefore not addressed to in
this fact sheet.
b) Heating and moisture increase

Mould growth in feed is accompanied by rising temperatures and moisture content
(FAO2, 1980). Mould species are known to produce water during their growth, which
enables other moulds to grow (Mills, 1989). A. candidus, can raise the moisture level of
the infested grain to 18% or higher.
Moulds raise the temperature of stored feed through their respiration. Development of
these moulds often occurs in pockets of increased moisture within bulks. Mould-induced
heating of stored feed attains temperatures of 55C and remains at this level for weeks.
The heating then either gradually subsides or passes into the next stage where
thermophilic moulds take over. These sometimes carry the temperature to 60C and may
be succeeded by other thermophilic microorganisms that carry it up to 75C (Mills, 1989).
The practice of storing bags of feed in large piles promotes mould activity. The "sweating"
of bags within a large stack is evidence of mould damage to the stored feed (FAO2,
1980).

c) Mustiness
Feed that is damaged by moulds tends to become lumpy. Feed grains suffer
discoloration while damaged maize turns a dark brown with some blackened kernels
being evident (FAO2, 1980). The musty smell reduces feed palatability and will reduce
dry matter intake of animals.
d) Loss of nutritional value

After field inoculation moulds utilize plant nutrients (Vieira, 2003) and decease the
nutritional value of feed (Bartov and Paster, 1986; Jin et al., 1999). Moulds are
considered the principal reason for the destruction of fats in grain during storage, which
occurs more rapidly than the destruction of other nutrients such as carbohydrates and
proteins (Bartov et al., 1982; Jen et al., 1999). Decreases in protein content and amino
acid levels are usually not measurable until the grain or feed is in an advanced stage of
deterioration. Mould growth may also result in lowered protein availability (Jen et al.,
1999). Heating, generated during mould growth may also affect the chemical composition
and nutritional value of feed (Sauer, 1988). During heating Maillard products may be
formed (Van Wagenberg et al., 2002). Maillard products are non-enzymatic polymers of
hemicellulose and soluble sugars with the amino groups of proteins. The nitrogen in
Maillard products is generally unavailable for digestion (Guerrero, 2006).
2. Transmission and likelihood of occurrence
Environment
There are several parameters that affect the growth of moulds in feed and food. Spores of
moulds have different germination parameters. Examples of parameters affecting the growth
of moulds are:
a) pH
A fairly wide range of pH levels will support mould growth, although they do not grow well
at extremely low or high pH levels (Whitlow and Hagler2, 2007). The pH range of growth
for moulds indicated by Ray (2004) is 1.5 to 9.0.
b) Moisture content (water activity [aw])
In general, moulds require less moisture than bacteria and yeasts. Moulds differ
considerably among themselves as to optimum water activity and range of water activity
for germination of spores. Water activity is the amount of water that is freely available
and not bound to feed and food molecules. The minimum water activity for spore
germination has been found to be as low as 0.62 for some moulds and as high as 0.93
for others. Each mould has an optimum of water activity and a range of water activity for
growth. The reduction of water activity below the optimum for a mould delays the
germination of the spores and reduces the growth rate (Lasztity, 2009). Whitlow and
Hagler2 (2007) state that water activity must generally be above 0.7 aw.
c) Oxygen and other gases
Moulds require free oxygen for growth (Lasztity, 2009). An oxygen level as low as 0.5%
can support mould growth; thus there can be pockets of adequate oxygen within silage
and stored high moisture grain storage, especially near the feed surfaces. High levels of
carbon dioxide can prevent mould growth even when oxygen is at levels high enough to
support mould growth (Whitlow and Hagler2, 2007).
d) Nutrient content
In order to grow and function normally, the fungi require the following: water, source of
energy, source of nitrogen, vitamins and related growth factors, and minerals.

e) Antimicrobials
Some antimicrobials, either natural constituents or chemically produced are known to
inhibit mould growth, e.g. coumarin (Ojala et al., 2000), potassium sorbate (Mehyar et al.,
2011).
f) Storage temperature
Most moulds grow well at ordinary room temperatures. The optimal temperature for most
moulds is between 25 and 30 C. Nevertheless it should be noted that some moulds
grow fairly well at temperatures of freezing or just above, and others can grow slowly at
sub-zero temperatures (Lasztity, 2009). Whitlow and Hagler2 (2007) indicate that
temperatures may range from -5 to 60 C.
g) Relative humidity
The relative humidity of the storage environment is important for the growth of moulds.
Generally the humidity level for spoilage moulds is above 70% (Mills, 1989).
h) Presence/activities of other microorganisms
Some microorganisms produce substances that are either inhibitory or lethal to others.
These include antibiotics, bacteriocins, hydrogen peroxide, and organic acids (such as
lactic acid). Ojala et al. (2000) observed antimicrobial properties of coumarin. Possible
mechanisms of interference are: competition for nutrients; competition for
attachment/adhesion sites; unfavourable alteration of the environment and/or
combinations of these (USDA, 2011). On the other hand Whitlow and Hagler2 (2007)
observed the opposite. While silage pH is generally low enough to prevent most mould
growth, yeasts are active at a lower pH and their activity can raise the pH to a point
conducive to mould growth.
Mould spores are present in the atmosphere (Sesartic and Dallafior, 2011). Airborne spores
can be carried long distances. Factors determining atmospheric transport and deposition are
spore size or shape, wind speed, temperature, atmospheric pressure or precipitation (Hirst et
al., 1967). Maus et al. (2001) studied the survival of mould spores in air filters and noticed
that after 5 days of incubation the number of studied spores remained stable in filters
exposed to air flow. Moulds and spores may also be present in dust (Schober, 1991; Hicks
et al., 2005).
Mould and mould spores can be present in surface water and in drinking water (Collins
and Willoughby, 1962; Warris et al., 2001; Hageskal et al., 2007). The presence of moulds in
drinking water seems to be related to the water source used and its treatment. Ground water
contains fewer moulds compared to surface water and spring/well water (Warris et al.,
2001; Hageskal et al., 2007; Pereir et al., 2009). Water treatment affects the presence of
moulds and mould spores. If water is stored in a manner that allows it to have contact with
ambient air (e.g., storage in a surface water reservoir), mould contamination will occur, and a
diversity of moulds can be recovered from outlets (e.g., taps, showers). On the contrary, in
closed water supply systems (e.g., storage of ground water in reservoirs where contact with
ambient air is prevented), mould contamination appears to be insignificant. The results of the
study by Warris et al. (2001) suggest that water purification procedures, such as chlorination,
do not eliminate mould spores.
Mould and mould spores are naturally present in soil (Mills, 1989; Rydjord et al., 2007). The
soil reservoir is the primary source of most mould infections in animals (Kahn, 2005). Mould
spores are known to survive in soil. Leong et al. (2006) studied the effects of water activity
and temperature on the survival of mould spores. Temperature and aw affect the survival of
these spores. At a low temperature spores survive for a longer period than at high
temperatures. The authors found that at 1C, spores survived for well over a year at 0.9 aw
and below.

Insects can physically damage stored crop (FAO3, 1994), making the crop accessible for
moulds. Insect can also spread moulds and spores. Mites feed on broken or damaged crop,
spread mould spores throughout the stored crop and into adjoining bins (Jacobs, 1988).
Moulds and mould spores are found in manure of various animal species (Lund, 1974;
Mc Carthy, et al., 2011). During composting the number of moulds is reduced however the
number of spores do not seem to be affected or even increased (Mc Carty et al., 2011). The
results of this assessment suggest that mould spores may be present in compost.
Mills (1989) mentions that moulds spores exist in harvesting equipment and within storage
structures and are gathered e.g. by the combine harvester and distributed among the crop.
Plants
Moulds play a role in the breakdown of leaves, wood and other plant debris and are as such
also present in these media (US-EPA, 2012). Mould spores are present in decaying plant
debris (Mills, 1989), lying ready to infect the growing plant in the field. In figure 1 the life
cycle of Phaeosphaeria nodorum is shown.
Figure 1. Life cycle of Phaeosphaeria nodorum (source: HGCA).
Cultivar differences exist and are of influence on mould susceptibility. The absence of a
protective sheaf makes sorghums and millets more susceptible than maize. However,
these crops do vary in their susceptibility due to the size and chemical constituents of the
grain. Larger, soft grain varieties are more susceptible than those with small, hard grains,
and the phenolic content of maize and sorghum is directly correlated with resistance to pest
damage; red and brown sorghum is less susceptible than white sorghum. Sorghum heads
that are open are more easily damaged by birds and so more prone to fungal invasion. Small
grain millet is much less damaged by insects than larger sorghum grains and both are less
susceptible than maize (FAO4, 2007).
Damaged crop is susceptible for mould infection (UoI, 2000), like broken grain kernels
(HGCA1, 2004).

There are various mould species and infestation of crops depends on the species involved,
but moulds are also known to infest a variety of crops, as shown in table 2.
2
Table 2. Some moulds related to crops (Christ, 1998; CCC, 2003; Ruhl, 2007; HGCA , 2012).
Crop Mould Related disease
Wheat Puccinia graminis f. tritici Stem rust
Phaeosphaeria nodorum Leaf and glume blotch
Blumeria graminis f. tritici Powdery mildew
Rye Puccinia graminis f. secalis Stem rust
Blumeria graminis f. secalis Powdery mildew
Barley Puccinia hordei Leaf rust
Blumeria graminis f. hordei Powdery mildew
Grasses Rhizoctonia solani Rhizoctonia stunt
Oats Ustilage segetum Covered smut
Blumeria graminis f. avenae Powdery mildew
Cereals (all) Cochliobolus sativus Foot rot
Alternaria spp. Black point
Rhizoctonia solani Rhizoctonia stunt
Maize Ustilago maydis Maize smut
Sphacelotheca reiliana Head smut
Cochliobolus heterostrophus Southern corn leaf blight
Soybean Septoria glycines Brown spot
Phytophthora sojae Phytophthora rot
Potato Phytophthora infestans Late blight
Rapeseed Rhizopus stolonifer Black mould rot
Because there are numerous moulds and any plant can be infested by moulds, no specific
mould if focussed on. Based on the worst-case scenario it is assumed that all untreated /
unprocessed products of vegetal origin can carry moulds and spores.
Moist products of vegetal origin are of risk especially when oxygen is available on the
surfaces, e.g. wet brewers grain, silage (cereal/grass) (Adams et al., 1993; Driehuis et
al., 2001; Keller et al., 2012). Postharvest spoilage of grass and maize silage by moulds is
dominated by Penicillium roqueforti, Aspergillus fumigatus and Zygomycetes (Storm et al.,
2008). Penicillium moulds are commonly found because they are acid tolerant and have a
low oxygen requirement. P. roqueforti is able to invade silage and is considered an indicator
of mould invasion. It destabilizes silage and results in an increased silage temperature, which
paves the way for other potentially pathogenic organisms. In the instable silage, toxin-
producing moulds and yeasts (Monascus ruber, Aspergillus fumigatus, Byssochlamys spp.)
may be found together with potentially pathogenic bacteria, such as Listeria monocytogenes
and Clostridia butyricum (Berge, 2011).
During storage mould and spores can infest any product of vegetal origin (untreated /
unprocessed or treated / processed) especially products high in moisture. Moulds and
spores can be present on the crop itself but also within the storage facility. Conditions
affecting the growth of moulds and spores are mentioned in the paragraph Environment,
above.
Mycelium of Aspergillus niger is a product consisting of the biomass of A. niger from the citric
acid fermentation that can be used in feed when dried. Drying should effectively kill residual
moulds. The A. niger strain is non-toxic and non-pathogenic and has a history of safe use in
production of food ingredients. The compositional data and the manufacturing process do not
give rise to concerns (EFSA1, 2010).
The growth of moulds can be inhibited or moulds can be killed by processing techniques
(e.g. heat treatment) or additives (e.g. organic acids). Concerning mould spores, this might
not be as effective compared to moulds.

Animals
Exposure:
Ruminants are relatively highly exposed due to the high proportion of forages they receive in
the ration (Morgavi et al., 2004). Mouldy feed however has been associated with reduced
palatability and reduced feed intake (Eduard, 2007).
Moulds and mould spores can also be inhaled from contaminated feed. This route of
exposure is not addressed to in this fact sheet.
No data were found concerning levels of exposure.
Absorption:
Lund (1974) studied the presence of moulds in the bovine rumen and found that the fungal
content of the rumen fluid seemed to be dependent on the diet of the animal, and no
particular fungal flora could be associated with the rumen. Spores are also found in the
rumen (Smith et al., 1987).
No data were found concerning the absorption of moulds as such.
Distribution:
White and Smith (1973) showed that A. fumigatus had a marked affinity for the (bovine)
placenta. They also suggest that the spores can reach the (bovine) placenta. No other data
were found concerning the distribution of moulds and mould spores after oral exposure.
Metabolism:
No data were found concerning mould and mould spore metabolism in animals.
Excretion:
Viable moulds and spores are excreted in faeces of ruminants (Lund, 1974; Smith et al.,
1987).
No data were found concerning the excretion of mould and mould spores in milk.
Humans via animal products

No data were found concerning the transfer of moulds and mould spores to products of
animal origin via feed. Due to the lack of data no specific products of animal origin can be
identified as of risk.
For these reasons the paragraphs humans will not be addressed to in the chapters:
transmission and likelihood of occurrence, diagnosing of poisoning, potential adverse
effects and severity of the potential adverse effects.
Not related to mould transfer from feed to a product of animal origin is the following. During
storage moulds may grow in products of animal origin. Specifically if these products are not
dried sufficiently as is reported in fish meal (FAO5, 1986; Windsor, 2001) and in meat (and
bone) meal (Husband, 1993).
Besides this it should be kept in mind that in food processing certain moulds are used
intentionally, e.g. the use of moulds in cheese processing. Mould contamination of products
of animal origin can also occur unintentionally after food processing. These moulds do not
originate from feed and are not addressed to in this fact sheet.
Humans
Exposure:
Not addressed to (see bold underlined text in chapter 2, section humans via animal
products).
Absorption:

products).
Distribution:
products).
Metabolism:
products).
Excretion:
products).
3) Diagnose of poisoning
Animals
Lund (1974) found that viable moulds were excreted in faeces of cattle. Toxins produced by
moulds can also been found in urine and faeces (Dailey et al., 1977) and possibly in blood.
Other body fluids or tissue may also be used.
Also history of exposure and analysis of feed can be included.
Humans
products).
4) Potential adverse effects

The potential effects of mycotoxins are not included in this fact sheet. These effects can be
severe. For more in formation about the potential adverse effects of mycotoxins is referred to
the fact sheets of specific mycotoxins available at GMP+ International.
Environment
No data were studied.
Animals
Ruminants are relatively highly exposed due to the high proportion of forages they receive in
the ration (Morgavi et al., 2004).
Mouldy feeds are less palatable and animals may eat less. This will lead to a reduction in
nutrient intake and so decrease weight gain and milk production. Even if mycotoxins are not
produced it has been estimated that the presence of mould alone will cause a loss in
performance of 5-10%. Mouldy feed may not be as digestible and the energy content will
therefore be reduced. Furthermore, the mould itself will use the protein, carbohydrate and fat
from the feed for its own development, reducing that available to the animal. Fat intake,
especially, is reduced in mouldy feed and energy availability may be reduced by as much as
10% (FAO4, 2007).
Scruggs and Blue (1994) reported two cases of herds with liver toxicity and
photosensitisation associated with feeding of mouldy alfalfa. A. fumigatus is associated with
bovine abortion (White and Smith, 1973).
Vesel et al. (1981) found that feeding maize silage with the mould P. roqueforti resulted in
loss of appetite, stoppage of rumen activity and gut inflammation. The mycotoxin analysed in
this silage was PR-toxin.

In Appendix I the potential adverse effects of moulds in animals are shown.
Humans
products).
5) Severity of the potential adverse effects

The severity of mycotoxins is not included in this fact sheet. The severity can be high. For
more in formation about the severity of mycotoxins is referred to the fact sheets of specific
mycotoxins available at GMP+ International.
The severity of mould contamination of food and / or feed materials is based on the worst
case scenario, as shown in table 3., and is based upon the potential adverse effects stated in
chapter 4 and Appendix I.
The severity of mould toxicosis in animals is classified as high because:

Mould: A. fumigatus is associated with bovine abortion (White and Smith, 1973).
The severity of mould and mould spores toxicosis in humans is not classified (see bold
underlined text in chapter 2, section humans via animal products).
Table 3. Severity of mould and mould spores

Severity
Low Medium High
Animals x
Humans Not classified
6) Legislation and standards

In Belgium Ovocom (2012) has stated mould action levels in feed. No other data concerning
mould legislation and standards were found.
7) Methods of analysis
Sampling: In GMP+ standard BA13 requirements for sampling are stated (GMP+3, 2013).
Analysis: There are various laboratory methods for the detection of moulds in food and feed.
Most methods detect moulds and yeast simultaneously.
The methods include different plating methods, fluorescent antibody techniques, direct
microscopical methods, ELISA techniques, ergosterol and chitin determinations, Howard
mould counts and colour measurements. The results of the study by Rabie et al. (1997)
indicate that the choice of method will, to some extent, depend on which mould species are
considered to be most important and, in particular, whether crop is freshly harvested or has
been stored. No single set of parameters will equally enumerate all species that occur in a
specific sample (Rabie et al., 1997).
A number of molecular methods based on immunological and genotypic techniques have
been developed for revealing the presence of undesirable microorganisms, including fungi, in
different food matrices (Van der Vossen and Hofstra, 1996; Li et al., 2000).
Some methods are highlighted below.

Culture and colony counting methods
Traditional culture techniques for detection of food-borne fungi (moulds and yeast) involve
the use of selective microbiological media, followed by the isolation of pure cultures and
finally the application of confirmatory tests. Although effective, these procedures are labour-
intensive and require several days (Bleve et al., 2003). In addition, viable bacterial strains in
the environment can enter a dormancy state where they become non-culturable (viable-but
non-culturable (VBNC)) which can subsequently lead to an underestimation of pathogen
numbers or a failure to isolate a pathogen from a contaminated sample (Xu et al., 1982;
Toze, 1999). VBNC cells are defined as those cells that have lost the ability to express genes
but may return to a culturable state, as is demonstrated in bacteria (Del Mar Lle et al.,
2000).
PCR-method
PCR (Polymerase Chain Reaction) is a molecular technique. Konietzny and Greiner (2003)
conclude that this technique may be applied to the screening of agricultural commodities for
the absence of mycotoxin producers. However, conventional PCR methods as culture and
colony counting methods do not distinguish among viable, VBNC and dead cells. The
presence of these cells limits the use of PCR for microbiological monitoring of food samples,
where metabolically injured or nonviable cells are generally present after the stresses
imposed during food processing (Bleve et al., 2003).
ELISA-method
Several ELISA methods (Enzyme-Linked ImmunoSorbent Assay) are available. Anand and
Rati (2006) describe the use of ELISA for monitoring of A. ochraceus in poultry feed.
8) Possible control measures

The HACCP system should be used to assess the risk of undesirable moulds in feed and
consequently the control measures needed to control this risk. All parts in the feed and food
chain should be included, starting from cultivation or husbandry.
Possible control measures are related to specific activities within each part of the feed and
food chain. However some control measures, e.g. concerning hygiene, are applicable in
each part of the feed and food chain.
In Appendix II possible general mould control measures are mentioned, applicable for
several part in the feed and food chain.
The 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:
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
undesirable moulds are summarised in table 4. There are no specific products of animal
origin of risk.
Table 4. Products with a higher risk to contain undesirable moulds or mould spores.
Any product in contact with contaminated manure, compost and plant debris;
Any product in contact with contaminated (harvesting) equipment that has not been
Products in general
sufficiently and regularly cleaned;

Any product in contact with contaminated water (e.g. spring water, surface water);
Any product in contact with contaminated soil. Also the adherence of contaminated soil to
vegetable surface is a risk.
Any product in contact with pests (e.g. insects, mites);
Any product in contact with contaminated air and/or dust (e.g. dust);
During storage mould and mould spores can infest any product of vegetal or animal origin
(untreated / unprocessed or treated / processed) especially products high in moisture.
Moulds and mould spores can be present on the product itself but also within the storage
facility.
Because there are numerous moulds and any plant can be infested by moulds, no specific mould
Specific products of
if focussed on. Based on the worst-case scenario it is assumed that all untreated / unprocessed
vegetal origin
products of vegetal origin can carry moulds and mould spores;

Plant debris;
Damaged crop (e.g. broken grain kernels);
Moist products of vegetal origin (e.g. wet brewers grain, silage (e.g. grass and maize);
Because of the absence of a protective sheaf sorghums and millets are susceptible to moulds.
However, these crops do vary in their susceptibility due to the size and chemical constituents of
4
the grain. Larger, soft grain varieties are also susceptible to moulds (FAO , 2007).
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10. Websites
1. http://www.hgca.com/document.aspx?fn=load&media_id=7326&publicationId=8742
2. http://www.hgca.com/document.aspx?fn=load&media_id=794&publicationId=1303
3. http://www.hgca.com/content.search/4/4/Site%20Search/Site%20Search/Redirect.mspx?
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4. http://www.hgca.com/content.search/4/4/Site%20Search/Site%20Search/Redirect.mspx?
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7. http://www.fao.org/docrep/T1838E/T1838E00.htm#Contents
8. http://www.fao.org/docrep/010/a1416e/a1416e00.htm

APPENDIX I Potential adverse effects of moulds and mould spores
1,2 1,2 1,2 1,2 2 2 2

Death Carcinogen Mutagen Reproductive Internal injury Neurological Immunological Organs
(physical
1,2
contamination)
Animals
X (mould) X (mould)
Humans
Not classified (see bold underlined test in chapter 2, section humans)
2 2 2 2
Dermal and Respiratory Musculo- Cardiovascular Gastrointestinal Hematological Endocrine Bodyweight
2 2
ocular skeletal
Animals
X (mould)
Humans
Not classified (see bold underlined test in chapter 2, section humans)
1
This potential adverse effect is classified as high severity for animals
2
This potential adverse effect is classified as high severity for humans

APPENDIX II Possible general control measures
In GMP+ B1 standard general control measures and requirements are stated which should
be complied (GMP+1, 2013). Additional recommendations for general control measures are
stated below in this appendix.
Personnel and other people
Each employee should be required to report to work in clean clothes and maintain
adequate, daily personal hygiene. It is highly recommended that employers provide clean
uniforms for use by the employees. The uniforms, footwear and other garments used in
the facility should stay on site and are not allowed to take home by the employees. If
reusable garments are used, the facility should provide for proper sanitizing (AFIA,
2010)3;
Persons suffering from communicable illness should not be allowed to work in the facility
(AFIA, 2010);
Every person must wash their hands with soap and water after each use of the restroom
and break facilities (AFIA, 2010);
Procedures should be in place to ensure that all visitors to the site, including staff,
contractors, transport operators and customers, are aware of the potential impact of their
actions on all aspects of product safety (AFIA, 2010);
Provide for sanitizing of footgear if deemed necessary and practical (AFIA, 2010);
Facilities
Facilities for employees can include the following (AFIA, 2010):

o Separate facilities for workers on the pre-processing side and workers on the post-
processing side of the plant;
o Adequate facilities for showering and dressing so as to minimize contamination carried
on clothing, shoes or the person;
o Adequate lavatory and toilet accommodations.
Sanitation and cleaning
The following applies to all areas:

Cleaning and sanitizing procedures should be appropriate for each specific piece of
equipment or area (AFIA, 2010);
Cleaning should include the following: the interior and exterior of production machinery
as well as ceilings, roof structure, wall cavities, ledges or rafters (AFIA, 2010). Dust
extractors / collectors and coolers should be cleaned regularly (FEDIOL, 2009)3;
Magnets should be cleaned at regular intervals to prevent feed from building up at these
points (AFIA, 2010);
Dry cleaning of spillages by sweeping and/or vacuuming is preferable to wet. If the use of
water is necessary, appropriate water temperatures and sanitizing steps should be
specified (and appropriate to the area being cleaned) (AFIA, 2010).
Settled dust should be removed using a vacuum cleaner rather than sweeping. Vacuum
cleaners should be dedicated to either pre- or post-process and should not be used in
other locations. Vacuum collection of dust is preferable to using compressed air, which
would increase air-borne dust. Employees should avoid using compressed air whenever
possible to remove dust from equipment or from clothing of personnel (AFIA, 2010);
3
These publications are Salmonella focussed. It assumed that the general Salmonella control measures are also
effective in controlling moulds.

Decontamination after mould detection
Once a mould contamination has been detected, decontamination should take place.
A procedure should be in place in case of the detection of mould.
In some instances, equipment may need to be partially disassembled to allow access for
cleaning (AFIA, 2010);
When partial disassembly is not a solution, dry flushing may be an option. The flush
should be isolated, discarded and not re-used within the facility (AFIA, 2010);
In case of a mould contamination in feed the cause of the contamination should be
determined. Appropriate measures should be taken to eradicate the cause. A procedure
should be in place describing how to handle mould contaminated products.
Pest control
An effective pest control program should be in place. Pests include rodents, birds,
insects, but also wild and domesticated animals (e.g. dogs and cats);
Keep the grounds surrounding the facility well drained and free of unnecessary
vegetation, such as weeds and high grass (AFIA, 2010);
Keep all areas within and around the plant free from accessible waste and trash;
Minimize dirt, dust, spilt feed and other organic materials and clean up spills promptly
(AFIA, 2010);
Unused equipment should be stored in a manner that eliminates pest infestation (AFIA,
2010);
Dust control and air flow
Dust is inherent in feed manufacturing. However, dust is a primary vehicle by which

microorganisms can be transmitted within the facility. Therefore, measures taken to control
and minimize dust are crucial to the success of any microorganism control program.
Ventilation within the production area should provide sufficient air exchange to prevent
accumulation of steam, condensation or dust and to remove contaminated air (AFIA,
2010);
Air drawn into the production area should be taken from areas likely to be clean (AFIA,
2010);
The control of air flow (and the dust contained in that air) should be designed so that
contamination does not spread from raw material areas into finished product areas of the
plant (AFIA, 2010);
Equipment has appropriate dust extraction capability (AFIA, 2010);
All processing and handling equipment should be constructed and installed in a manner
to minimize leakage, spillage and dust accumulation (AFIA, 2010);
Dust collection systems should be adequate to control dust and to aid in keeping the
plant clean (AFIA, 2010);
Dust extractors / collectors should be cleaned regularly and condensation should be
prevented where possible (FEDIOL, 2009);
Remove settled dust with a vacuum cleaner equipped with a highly efficient filter rather
than sweeping. Avoid using compressed air to remove dust from equipment or from
clothing or shoes of persons (AFIA, 2010);
Dust from cleaning, maintenance and overhaul of equipment such as extractors,
cyclones and filters, should not re-enter the finished meal stream (FEDIOL, 2009), they
should be disposed of (AFIA, 2010);

Moisture control
The areas should be designed for wet weather operation, so that the loading and
unloading of feed occurs without significant water damage to the feed (AFIA, 2010);
Roofs, ceilings and walls should be leak-proof. Construct storage-area walls and floors in
such a manner as to keep out moisture (AFIA, 2010);
Keep feed dry at all times. (AFIA, 2010);
Avoid or correct conditions conducive to the formation of condensation in buildings,
equipment (AFIA, 2010) and storage and transport entities;
Pellet coolers should be operated in a manner to prevent condensation on interior
surfaces that encourage mould growth and should be regularly monitored for
contamination (AFIA, 2010);
Any wet material should be disposed of as waste or recycled through effective heat-
processing step(s) (AFIA, 2010).

APPENDIX III Mould control measures: cultivation, harvesting, processing, transport
and storage of feed on farm.
Specific requirements and control measures related to feed ingredient cultivation are stated
in the GMP+ B6 standard (GMP+2, 2013). This standard should be complied with. Additional
recommendations for general control measures are stated below in this appendix.
Cultivation, harvesting, processing, transport and storage on farm
Use the HACCP system to assess the risk of mould and mould spores in feed. Besides
others, the following items should be addressed:
Cultivation: Timing of the production cycle. Planting must take place at the recommended
time to avoid problems caused by the crop maturing too early or more particularly, too
late, during periods of prolonged rainfall. If plants mature when the relative humidity is
high or whilst it is raining they will be prone to invasion by moulds, as well as being
subject to pre-harvest damage by storage insect pests, which themselves will help with
the diffusion and multiplication of mould spores (Bruns, 2003; FAO4, 2007);
Cultivation: Apply crop rotation (FAO4, 2007);
Cultivation: Remove crop debris or apply tillage e.g. by ploughing, by burning stubbles.
Crop debris should not remain in the surface layer of the soil (PDV2, 2000; FAO4, 2007);
Cultivation: Use disease and mould-free sowing seed: buy and plant certified seed
(PDV2, 2000; FAO4, 2007);
Cultivation: Do not used by insects infested sowing seeds (FAO4, 2007);
Cultivation: Plant seed varieties that are mould and mycotoxin resistant (FAO4, 2007) or
less susceptible for mould and mycotoxin contamination (PDV2, 2000);
Cultivation of cereals: Use long straw varieties. Compared to short straw varieties the
ears of long straw varieties are less easy infected via spattered water from the ground
during e.g. raining. Long straw varieties also dry more quickly after precipitation (PDV2,
2000);
Cultivation: Plant seed pest resistant varieties. Varieties with resistance to insects and to
viral and bacterial disease are available and should be used. Also keep in mind that
cultivar differences exist for many pre-harvest factors including pest resistance, drought
tolerance, stalk strength and husk cover. There are significant differences in keeping
qualities between different crop varieties. Local varieties of maize, which have small
cobs with tight, elongated husks that completely enclose the kernels, are relatively
resistant to insect attack and therefore less susceptible to fungi. The sheathing leaves
provide a physical barrier to entry. Composite and more particularly, hybrid varieties tend
to have much poorer husk cover, the leaves fail to fully enclose the larger cobs of these
varieties. Consequently, they are much more prone to damage by insects and moulds
and extra care must be taken during crop maturation and drying to ensure that these
varieties are not invaded by pests and diseases (in practice, without artificial drying, this
is difficult to achieve where rains are prolonged). Conversely, the tight husks of small
local varieties restrict water loss and slow drying (FAO4, 2007);
Cultivation: Prevent drought stress caused by the lack of water. Drought stress is known
to facilitate attack by mould. Where irrigation systems function, farmers should try to
supplement local rainfall to avoid drought stress (FAO4, 2007);
Cultivation: Weeding will help the plant fight against insect pests which damage the
growing plant as does the application of insecticides (FAO4, 2007);
Cultivation: When necessary apply effective pesticides to control fungi and insects;
Cultivation: Avoid plant stress by providing sufficient plant nutrients. The application of
fertilizer should be timely and in the correct quantity (FAO4, 2007). The dosage of excess
fertilizer leas to an increase level of nutrients in the plant, e.g. nitrogen. This increases
the risk of mould infection. The plant grows so quickly resulting in a weak plant. Excess
fertilizer encourages more leaf on the plant so that the crop dries less quickly. The plant
is more attractive to insects because of a higher protein level and ripening is slower

(PDV2, 2000);
Harvesting: Harvesting machinery should be clean before harvesting begins (Bruns,
2003);
Harvesting: Properly setting of harvesting machinery and driving harvesting machinery at
ground speeds that do not overload the threshing mechanism in order to minimize kernel
breakage (Bruns, 2003);
Harvesting: Plentiful amounts of air should be blown through the crop during threshing to
remove foreign matter, weed seed, and light chaffy grain that may be infected with
moulds and contaminated with mycotoxins (Bruns, 2003);
Harvesting: Harvest as quickly as possible (FAO4, 2007). Harvest crop when the crop is
properly ripened. The correct moisture content at the moment of harvesting. It may be
necessary to dry the harvested crop (PDV2, 2000);
Harvesting: Avoid harvesting of wet / moist crop and when needed dry the crop to a safe
storage moisture level (FAO4, 2007);
Harvesting: Avoid harvesting lodged or fallen material, contact with soil can increase
mould and mycotoxin contamination (Whitlow, 2006; EC, 2006);
Harvesting: Avoid field drying (FAO4, 2007);
Drying: See Appendix VII Processing;
Transport and storage: Avoid carry-over of mould infected crop via transport and storage
entities to other feed transported by these transport and storage entities (e.g. clean
transport and storage entities). For more information see Appendices V and VI.
Mould monitoring programs:

Implement various mould-monitoring programs, as stated hereafter. In GMP+ Appendix

BA13 a sampling protocol is stated for microbiological examination (GMP+3, 2013). Several
types of monitoring programs are suggested:
Implement a mould monitoring program, if applicable, meeting the requirements stated in
applicable cultivation quality systems (e.g. Eurep-GAP). Concerning mould monitoring
programs for the transport and storage of feed is referred to appendices X and XI
respectively. For specific information concerning mould monitoring in cultivation, see
below;
Implement a mould monitoring program on the following products, if applicable, which
were determined of risk by the HACCP study: e.g. sowing seeds, irrigation water, soil,
fertilizer, manure, compost materials, residues and spills in specific areas, silage, moist
feed;
Implement a mould monitoring at critical points in cultivation, as is determined using the
HACCP system. Also include machinery and equipment in the monitoring program. A
representative number of samples should be taken and examined from the critical points.
Critical points where samples can be taken are e.g.:
o Dust from ledges inside and the top of feed silos / bins;
o Machinery and equipment in direct contact with feed;
In the event of mould (contamination corrective measures will be taken immediately, see
section Decontamination after mould detection in Appendix II.

APPENDIX IV Mould control measures: husbandry.
Husbandry
Use the HACCP system to assess the risk of moulds in feed during husbandry. Besides
Food producing animals are kept and certified according to the standards stated in a
quality program (e.g. IKB, KKM, QM-Milch etc.) addressing and controlling hygiene and
microorganisms;
Home mixing of feed: the home mixing of feed should be certified in compliance with a
quality program addressing hygiene and microorganisms;
Home mixing of feed: cleaning procedures of vehicles and equipment used for milling
activities should be in place;
In case of buying feed at third parties, suppliers can be selected based on their
implementation of programs for the control of microorganisms or hygiene;
Feed fed should be dry (if applicable), without organoleptic abnormalities (e.g. normal
smell and temperature, absence of mould);
Feed storage systems should operate on a first-in-first-out system (AFMA, 2003);
Water: food-producing animals should have access to clean drinking water (e.g. mind
when well water possibly contaminated with manure / excrements of pests and wild and
domesticated animals);
Feeding system: avoid stepping on feed and feeding feed on walkways;
Feeding system: avoid the possibility of manure, litter, water, soil, slurry or sewage to get
in contact with feed;
Feeding system: leave no residues in the feeding system after feeding;
Feeding systems: open systems attract birds, rodents, insects and pets. A pest control
program should be in place: see section Pest control. For possible control measures
concerning storage of feed in general: see Appendix VI Storage;
Feeding systems: cleaning procedures should be in place for feeding systems, including
equipment and machinery used.
Pig feeding systems with their complex of troughs, hoppers and pipes have been shown
to be particularly difficult to clean and disinfect effectively because of inaccessible
surfaces and pooling of wash water (EFSA2, 2008);
If wet cleaning is needed, any residual water after wet cleaning should be removed and
all contact surfaces should be dry
Mould monitoring programs: husbandry

Implement a mould monitoring program, if applicable, meeting the requirements stated in
applicable husbandry quality systems (e.g. IKB, KKM, QM-Milch). In case of home mixing
of feed or drying of feed, the monitoring program on feed in Appendix XII is referred to.
Concerning mould monitoring programs for the transport and storage of feed is referred
to appendices XI and XI respectively. For specific information concerning mould
monitoring in husbandry, see below;
Implement a mould monitoring program on products which were determined of risk by the
HACCP study: e.g. feed (incl. on farm produced products of vegetal origin, compound
feed, silages, milk replacers, additives, premixtures, water (e.g. drinking water, well
water), residues and spills in specific areas (e.g. in trench silo, in feeding system),
compost and manure;
Implement a mould monitoring at critical points in husbandry, as is determined using the
HACCP system. Also include machinery and equipment in the monitoring program. A

Mould monitoring programs: husbandry
representative number of samples should be taken and examined from the critical points.
Critical points where samples can be taken are e.g.:
o Dust from ledges inside and the top of feed silos / bins;
o Feeding systems;
o Troughs (incl. milking parlour);
o Buckets and teats (e.g. used for milk replacer feeding);
o Machinery used for mixing feed or other machinery in direct contact with feed;
o Milking equipment;
o Equipment in direct contact with feed;
In the event of mould contamination corrective measures will be taken immediately, see

APPENDIX V Mould control measures: transport of feed.
be complied with (GMP+1, 2013). Specific requirements and control measures related to
transport are stated in various GMP+ standards (GMP+4,5,6,7,8,9, 2013). The applicable GMP+
standard depends on the means of transport. The applicable standard should be complied
with. Additional recommendations for general control measures are stated below in this
appendix.
Transport
Use the HACCP system to assess the risk of moulds in the feed during transport. Besides
Mind the temperature of feed at loading. The temperature should be low enough to
ensure that no condensation and heating will occur;
If covers are used during transport, they must be maintained in a clean condition by
being cleaned, sanitised and dried regularly (EFSA2, 2008);
Avoid damage caused by pests (PDV2, 2000);
If transport can be made airtight, fumigate if pests are present (FAO1, 2007);
Avoid temperature fluctuations (PDV2, 2000);
Fungicides or approved preservatives can be used to reduce mould contamination within
the transported lot (PDV2, 2000).
Mould monitoring programs: transport

Implement a mould monitoring program on feed that is determined of risk by the HACCP
study, also residues and spills in specific areas;
Implement a mould monitoring program inline at critical points in transporting feed, as is
determined using the HACCP system. Also include machinery and processing equipment
in the monitoring program. A representative number of samples should be taken and
examined from the critical points. Critical points where samples can be taken are e.g.:
o Surfaces of e.g. trailers, containers and holds;
o Covers;
o Equipment used during loading and unloading;
o Dust bag.

APPENDIX VI Mould control measures: storage of feed.
be complied with (GMP+1, 2013). Specific requirements and control measures related to
transport are stated in the GMP+ B5 standard (GMP+10, 2013). Additional recommendations
for general control measures are stated below in this appendix.
Storage
Use the HACCP system to assess the risk of mould in the feed during storage and
transhipment. Besides others, the following items should be addressed:
Clean the harvested crop by removing fines (damaged kernels and small pieces of
foreign material (Whitlow, 2006);
Careful and rapid drying of feed, if the feed is too moist. See Appendix VII Processing.
Examples of maximum moisture content for various crops are given in table VI.1.
Table VI.1 maximum moisture content for various crops long-term storage (more than one crop
1,2,3,4,5 4
year) (White and Toman, 1994; CA , 1995; DAFWA, 2005; HGCA , 2005; ISU, 2008).
Product % Moisture
Beans 15
Chick peas 14
Cow peas 15
Lentils 15
Maize 15,5
Oats 14
Peas 15
Pulses 14
Rape seed 12
Sorghum 14-14,5
Soy bean 11-12
Wheat 14-14,5
Frequently monitor temperature and moisture at several depths until these have
stabilised for long-term storage. Crops should than be monitored regularly (FSA, 2007).
In case the temperature or moisture content increases above an acceptable level, drying
or ventilating should be considered. See Appendix VII Processing;
Insect and mite populations should be monitored using traps and/or grain sieving (FSA,
2007);
If authorised, top dressings may be used to control mites as long as the moisture content
in the bulk is low enough (HCGA4, 2005);
If authorised, insecticides may be used to control insects as long as the moisture content
in the bulk is low enough (HCGA4, 2005);
Application of storage fungicides (White and Toman, 1994);
A pest control program should be in place: see section Pest control;
Clean stored crop (e.g. with sieves) in order to remove damaged crop, e.g. cereal kernels
(AFMA, 2003; FAO4, 2007);
Storage entities should have adequate head space and ventilation to avoid re-absorption
of moisture by (dried) grains (FSA, 2007);
Perforations in ducts/flooring should be kept clean to ensure adequate airflow (FSA,
2007);
Store bags on a platform raised above the floor (FAO4, 2007);
Storage systems should operate on a first-in-first-out system (AFMA, 2003);
Avoid storing feed in bins/silos exposed to sunlight. Feed in bins/silos exposed to sunlight
is prone to moisture migration. The feed thus tends to cake on the bin walls, particularly
in corrugated bins. This feed becomes mouldy and then continually seeds fresh feed with
spores (AFMA, 2003). Bins/silos should be situated such to avoid moisture migration
caused by sunlight.
Regular cleaning and treatment with, if authorised, a mould inhibitor of the conveyor
system (e.g. in dry powder form) will reduce or prevent mould colonization (AFMA, 2003).

Mould monitoring programs: storage

types of monitoring programs are suggested: on feed and on critical points in the storage of
feed, as stated below:
Implement a mould monitoring program on feed that is determined of risk by the HACCP
study, also residues and spills in specific;
Implement a mould monitoring program inline at critical points in storing feed, as is
examined from the critical points. Critical points where samples can be taken are e.g.:
o The unloading pit / dumping pit for feed, e.g. dust from auger system;
o Dust from augers, conveyer belts, elevators;
o Dust from air aspiration cyclone system;
o Dust from vacuum cleaners;
o Machinery and equipment in direct contact with feed;
o Dust from cleaning, maintenance and overhaul of equipment;
o Dust from ledges inside of feed silos / bins, or dust from sieves;
o If applicable: inside of coolers where condensation sites are possible e.g. the top of
the pellet cooler;
o If applicable: dust from coolers, taken below coolers or on associated framework,
ledges etc.;

APPENDIX VII Mould control measures: processing of feed.
In GMP+ B1 and B2 standard general control measures and requirements are stated which
should be complied with (GMP+1,11, 2012). Additional recommendations for general control
measures are stated below in this appendix.
Purchasing and receiving feed
In case risk products are used, as determined by using the HACCP system, a procedure
should be in place to assure safe use as feed (e.g. analysis of moulds prior unloading);
Do not use risk products as feed for susceptible animals, as determined by using the
HACCP system, unless it is assured that the occurrence of moulds is at an acceptable
safe level and under control;
Verify if the supplier / producer of feed (also include premixes, additives etc.) has
assessed the hazard and risk of moulds;
Selecting of the suppliers based on their implementation of programs for the control of
moulds and hygiene;
Feed arriving at the production site should be subjected to arrival inspections and include
the following:
o Inspection of documentation, invoices and seals (if applicable);
o Assessment of transport entities with respect to maintenance, sanitation, cleanliness
and, if applicable, covered transport entities;
o Verification of ingredient identity (e.g. by documents, visual inspection);
o Inspection of ingredient for quality indicators, including the following:
Visible evidence of water damage;
Visible faecal or vermin contamination;
Temperature check;
Aroma;
The receiving area should be designed for wet weather operation, so that the unloading
of ingredients occurs without significant water damage to the ingredients.
Drying and ventilating

Dry up to a moist content unfavourable for the growth of moulds, however avoid cracking
of kernels during drying;
Careful and rapid drying or ventilating of feed, if the feed is too moist or the temperature
is to high. Examples of maximum moisture content for various crops are given in table
VII.1
Table VII.1 maximum moisture content for various crops long-term storage (more than one crop
1,2,3,4,5 4
year) (White and Toman, 1994; CA , 1995; DAFWA, 2005; HGCA , 2005; ISU, 2008).
Product % Moisture
Beans 15
Chick peas 14
Cow peas 15
Lentils 15
Maize 15,5
Oats 14
Peas 15
Pulses 14
Rape seed 12
Sorghum 14-14,5
Soy bean 11-12
Wheat 14-14,5
o Hot-air drying: drying temperature should be set according to manufacturers
instructions (FSA, 2007). Care should be taken not to dry to too low moisture
content by high-temperature drying. This can cause stress cracking of kernels
and results in greater amounts of mechanical damage each time the product is
handled which makes it more susceptible for mould infestation (White and Toman,

1994);
o Fan drying: an airflow of at least 180 m3/hour/tonne is recommended during
drying (FSA, 2007);
o Fan drying: cereal depth of the drying zone should be based on fan capacity and
availability of added heat. Grain depth should be reduced when cereals entering
store has high moisture content (>20% moisture) (FSA, 2007). Where for the
drying of rapeseed cereal driers are used, the bed depth should be reduced by
50% to 70%, depending on the output (HCGA4, 2005);
o The cereal surface of the drying zone should be levelled so that cereal depth is
uniform (FSA, 2007);
o Cereal stirrers can be used to increase airflow and mix cereals from different
drying zones (FSA, 2007).
The dried feed should be cooled up to a temperature as low to prevent condensation and
heating during storage:
o Hot-air drying: cereals should be cooled back to near-ambient temperature (FSA,
2007);
o Fan drying: cool night-time temperatures should be used to cool feed. Cereal is
quickly cooled when ambient air temperature is at least 5C below cereal
temperature (FDSA, 2007)
o Fan drying: an airflow of 10 m3/hour/tonne is recommended for cooling cereals
(FSA, 2007);
o Where for the cooling of rapeseed cereal coolers are used, the bed depth should
be reduced by 50% to 70%, depending on the output (HCGA4, 2005);
o Feed should be continuously cooled (FSA, 2007).
Conditioning, heat or other technological treatment (e.g. pelleting, expanding, extrusion)
The use of high temperatures to accomplish pasteurisation during processing is based on the
destructive effects of time and temperature on microorganisms. Microbiologists have
identified at least 11 factors or parameters of microorganisms and their environment that can
affect heat destruction. These factors include moisture or water activity, fat levels, presence
of salts, presence of carbohydrates, pH, protein content, number of organisms, age of
organisms, inhibitory compounds, and time and temperature history. Despite the influence
these factors can have on the resistance of microorganisms to heat, thermal destruction
during the processing steps of pelleting or extrusion is the most critical control step for
destruction or reduction of microorganisms (AFIA, 2010)4.
Microbial populations are not killed instantly upon exposure to heat, moisture and
pressure. The effective heat treatment should be determined at the individual plants.
Conditioning times in conventional equipment generally range from a few seconds to
several minutes, depending on the equipment involved. The pelleting process can also
be augmented with the use of expanders (Jones4, 2011). The EFSA2 (2008) also
includes conditioning prior to pelleting or expanding as usual processing procedures
performed in most feed mills when heat treated feed is manufactured.
Besides heat treatment other method may be effective in killing moulds. These methods
should be validated and verified by experimental studies;
The effectiveness of the treatment in killing moulds should be verified.
After heat treatment, cooling and drying should occur as rapidly as possible to reduce
condensation in processing and conveying equipment, storage containers, packaging
equipment and in the distribution of finished products (AFIA, 2010);
Thermocouples or temperature monitoring devices should be strategically placed in the
process flow to monitor the actual temperature (AFIA, 2010);
4
These publications are Salmonella focussed. It is assumed that the general Salmonella control measures are
also effective in controlling moulds.

Coolers and dryers
After heat processing, cooling and drying should occur as rapidly as possible to reduce
condensation in processing and conveying equipment, storage entities, packaging
equipment and in the distribution of finished products (AFIA, 2010);
Packaging
In case of bagging, new bags (incl. big bags) are recommended for finished product
packaging. Previously used bags should be sanitised before used for packaging (AFIA,
2010).
Mould monitoring programs: processing

BA13 a sampling protocol is stated for microbiological examination (GMP+3, 2012). Three
types of monitoring programs are suggested: on feed and on critical points in the storage of
feed, as stated below:
Implement a mould monitoring program on the following ingredients and intermediate
products, which were determined of risk by the HACCP study: e.g. feed materials,
processing aids, water, pressed meal before and after heat treatment, residues and spills
in specific areas;
Implement a mould monitoring program inline at critical points in processing feed, as is
examined from the critical points. Critical points, besides the ones indicated in the GMP+
standards, where samples can be taken are e.g.:
o The unloading pit / dumping pit for feed (SVA, 2011)4, e.g. dust from auger system
(DEFRA, 2009)4;
o Dust from ledges inside of feed silos / bins, or dust from sieves (DEFRA, 2009);
o Dust from hammer mills (DEFRA, 2009);
o Inside of coolers where condensation sites are possible e.g. the top of the pellet
cooler (SVA, 2011);
o Dust from coolers, taken below coolers or on associated framework, ledges etc.
(DEFRA, 2009);
o Dust from air aspiration cyclone system. Unless this is not accessible vacuumed or
swept dust could be used (DEFRA, 2009);
o Dust from vacuum cleaners (DEFRA, 2009);
o Dust from pellet shakers (DEFRA, 2009);
o Dust from augers, conveyer belts, elevators;
Implement a mould monitoring program on feed (e.g. compound feed, feed ingredients
directly delivered to farmers (e.g. cereals)), with a higher frequency of analysis in feed of
high(er) risk (based on the HACCP study);


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Fact sheet:

Moulds and yeasts

GMP+ International B.V.

All rights reserved. The information in this publication may

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

Summary of GMP+ products standards for the animal feed sector

Fact sheet: Moulds and yeasts 2 / 34

Name: Moulds and yeasts

Control measures: - (Locked) closed storage

Fact sheet: Moulds and yeasts 3 / 34

Feed materials 12% moisture content or aw- -

Fact sheet: Moulds and yeasts 4 / 34

This fact sheet focuses on mould as undesirable substance.

1. Nature, history and prevalence of moulds

Figure 1. Mouldy grain (source: ukmalt.com)

Fact sheet: Moulds and yeasts 5 / 34

Detrimental effects of moulds on feed are:

a) Mycotoxin and MVOCs production

Table 1. Fungi commonly reported in feed or food.

b) Heating and moisture increase

Fact sheet: Moulds and yeasts 6 / 34

d) Loss of nutritional value

2. Transmission and likelihood of occurrence

Fact sheet: Moulds and yeasts 7 / 34

Fact sheet: Moulds and yeasts 8 / 34

Figure 1. Life cycle of Phaeosphaeria nodorum (source: HGCA).

Fact sheet: Moulds and yeasts 9 / 34

Fact sheet: Moulds and yeasts 10 / 34

Humans via animal products

Fact sheet: Moulds and yeasts 11 / 34

4) Potential adverse effects

Fact sheet: Moulds and yeasts 12 / 34

5) Severity of the potential adverse effects

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

Table 3. Severity of mould and mould spores

6) Legislation and standards

Fact sheet: Moulds and yeasts 13 / 34

8) Possible control measures

Fact sheet: Moulds and yeasts 14 / 34

sufficiently and regularly cleaned;

products of vegetal origin can carry moulds and mould spores;

Fact sheet: Moulds and yeasts 15 / 34

Fact sheet: Moulds and yeasts 16 / 34

Fact sheet: Moulds and yeasts 17 / 34

Fact sheet: Moulds and yeasts 18 / 34

Fact sheet: Moulds and yeasts 19 / 34

Fact sheet: Moulds and yeasts 20 / 34

1,2 1,2 1,2 1,2 2 2 2

Fact sheet: Moulds and yeasts 21 / 34

Personnel and other people

Facilities for employees can include the following (AFIA, 2010):

Sanitation and cleaning

The following applies to all areas:

Fact sheet: Moulds and yeasts 22 / 34

Dust control and air flow

Dust is inherent in feed manufacturing. However, dust is a primary vehicle by which

Fact sheet: Moulds and yeasts 23 / 34

Fact sheet: Moulds and yeasts 24 / 34

Cultivation, harvesting, processing, transport and storage on farm

Fact sheet: Moulds and yeasts 25 / 34

Mould monitoring programs:

Implement various mould-monitoring programs, as stated hereafter. In GMP+ Appendix

Fact sheet: Moulds and yeasts 26 / 34

Mould monitoring programs: husbandry