General Review of Mycotoxins

Dr.Kedar Karki
 Mycotoxins

• Mycotoxins are secondary metabolites produced by fungi

present in feed.
• Mycotoxin's production depends on fungus specie and
strand, plant specie, environmental moisture and
temperature, presence of pests, etc.
• Mycotoxins cause damages in feed quality.
• Their incidence depends on geographical area and
season.
• Mycotoxins are toxic: they produce mycotoxicosis and
drop of performance.
• Their presence in feed can be reduced by applying
Hazard Analysis and Critical Control Points.
 Mycotoxin toxicity
Main factors that influence toxicity of
mycotoxins are:
• Bioavailability.
• Combined effects between several mycotoxins.
• Amount of mycotoxins consumed. Continuous or
intermittent ingestion of the contaminated feed.
• Animal weight, age, physiological and health
status.
 Mycotoxicosis

• Mycotoxicosis (1962, Forgacs and Carl): host's

intoxication as a result of ingestion of a toxic
substance of fungal origin.Some cases show
evident symptoms that can be easily associated
to mycotoxicosis.
• In the other hand, subclinical mycotoxicosis is
only recognizable by drop of performance and
health status.
• Susceptibility to mycotoxicosis depends on
animal specie, age, sex and coexistence with
other illness.
 Mycotoxicosis can cause:

• NUTRITIONAL • HEALTH ASPECTS

ASPECTS • Mycotoxicosis typical
• Feed consume of every mycotoxin.
decrease.
• Immunosupression:
• Nutrient absorption arise of other
decrease. pathologies.
 MYCOTOXINS CLASSIFICATION
AND MODE OF ACTION
• Lots of described mycotoxins.

• Varied molecular weights and

structures: difficult to classify.

• Mycotoxins persist in food

• They keep associated to chain.
fungus or substrate.

• Many of them are stable to

chemical/physical treatments.
 Classification according to
pathology
Hepatotoxins: sporidesmine, aflatoxins, luteoskirin, cycloclorotin,
rubratoxins, sterigmatocistin.
Nephrotoxins: ochratoxin, citrinin.
Neurotoxins: penitrem A, patulin, fumonisins, citreoviridin.
Toxins of intestinal tract: trichotecenes, T-2 toxin, deoxynivalenol
(Don, Vomitoxin), HT2 toxin, fusarenone.
Steroidal; strogenic (Zearalenone), D vitamin analogous
Haemorrhagic and circulatory toxins: Ergot alkaloids, aflatoxins.
Classification according to
chemical structure
 Chemical structure

• Chemical structure determines:

• Mycotoxin's mode of action.
• Mycotoxin's method of detoxification
Chemical structure and mode of
action Mode of action:
• specific biochemical interaction through
which a substance produces its biological
effect.
• In order to achieve a biological effect, an
interaction with a receptor is essential.
Chemical structure and mode of
action
• Chemical groups of the receptor must
interact with chemical groups of the
substance, that is, chemical structures in
the binding point must be complementary.
 KEY-LOCK MODEL

Mycotoxins with shared chemical structure

may interact with the same receptors an thus have an alike biological effect.
Chemical structure and mode of
action

• Into practice T2 toxin, HT2 toxin,

deoxynivalenol, nivalenol have a
sesquiterpene group in their structure and
they all have necrotic effects on mucous
membranes.
Chemical structure and mode of
action
• B1, B2, G1, G2 aflatoxins, sterigmatocistin
have cumarinic group in their structure
and they all have haemorrhagic effects
alike anticoagulant active principles used
in human pharmacology: warfarine,
acenocumarol. These active principles
also show a cumarinic structure.
Aflatoxin B1
MYCOTOXIN ANALYSIS IN FEED
MANUFACTURING
• Decision making in raw material
purchasing
 Usual methods of analysis

• Thin layer Chromatography (TLC).

• Liquid Chromatography (HPLC).
• Gas Chromatography-Mass spectrometry
(GC-MS).
• Immunoassay(ELISA).
Maximum allowed concentration

• Gradually more countries legislate about mycotoxin limits

in fodder and raw materials destined to animal nutrition.

Maximum concentrations are set depending on:

• Mycotoxin's toxicity
• Animal specie sensitivity and age
• Fungi load characteristic of plant specie
• Availability and limits of analysis method.
• Maximum concentration for each mycotoxin depends on
every country.
Maximum allowed concentration
• Maximum concentration depends on
animal specie and age and on raw
material or fodder.
 INTERPRETATION OF RESULTS

• Interpretation of results

Considering the results obtained in the lab, decisions are

made taking into account:
• Concentration of each mycotoxin (individual effect).
• Concentration of all mycotoxins analysed as a whole
(combined effects).
• Possible bias of analysis.
• Presence of non-analysed mycotoxins.
COMBINED EFFECTS
 There are more possibilities of
finding combined effects in
mycotoxins...
• With similar molecular structure.
Synthesized by the same fungal strand or
specie.
Synthesized by the same fungal family.
 The appearance of combined
effects depends on:
• Concentration of each mycotoxin.
• Animal sensitivity (specie, age, health
status).
 Additive effect

• Combined effect of A and B mycotoxins is

equal to the addition of the effect of each
mycotoxins.
 Additive effect

• Aflatoxins + Deoxynivalenol
Poultry: decrease in proventriculus weight, increase of
DHL enzyme, indicator of tissue damage.

• Aflatoxins + Cyclopiazonic acid

Poultry: growth decrease.
Pigs: feed intake and growth decrease; inflammation and
necrosis of the gastrointestinal tract. hepatotoxicity.
 Additive effect

• Aflatoxins + Diacetoxyscirpenol
Pig: Weight and growth decrease, alteration of
blood parameters that indicates hepathotoxicity.

• Aflatoxins + Moniliformin
Poultry: weight and efficiency decrease.
Increase of heart's relative weight. Biochemical
parameters indicate nephro and
 Additive effect

• Citrinin + ochratoxin A
Pig: nephrotoxicity. They affect transport of several molecules and
protein synthesis.

• Fusaric
Poultry: feed consumption and growth decrease. Nephro and
cardiotoxicity.

• Ochratoxin A + T-2 toxin

Poultry: Weight decrease, increase of kidney, liver, proventriculus
and gizzard relative weight. Nephro and hepatotoxicity.
 Additive effect

• Fumonisin B1 + Diacetoxyscirpenol
Turkey: Weight decrease. Hepatotoxicity.

• Fumonisin B1 + T-2 toxin

Poultry: weight and efficiency decrease. Nephro and
hepatotoxicity.

• Deoxynivalenol + Moliniformine
Turkey: weight of kidney and heart increases. Tissular
damage in myocardium.
 Synergic effect

•
Combined effect of A and B mycotoxins is
higher than the addition of the effect of
each mycotoxin.
 Synergic effect

• Aflatoxins + Ochratoxin A
Poultry: Weight decrease, mortality increase.
Hepatotoxicity and severe nephrotoxicity.

• Aflatoxins + Toxin T-2 Very important

because of its prevalence
Poultry: weight decrease, increase of kidney,
gizzard and heart relative weight; decrease of
the medium corpuscular volume and of the
potassium plasma levels.
 Synergic effect

• Deoxynivalenol + Fumonisin B1
Pigs: great weight decrease.

• Deoxynivalenol + Zearalenone
Pigs: theratogenesis in piglets.
 Antagonistic effects

• Combined effect of A and B mycotoxins is

less than the addition of the effect of each
mycotoxin. (but higher than the effect of
each mycotoxin separately).
 Antagonistic effects

• Citrinin + ochratoxin A
Poultry: the presence of these mycotoxins
together reduces the toxic effects of the
mycotoxins separately (growth and water
consumption decrease).

• Aflatoxins + diacetoxyscirpenol
Poultry: the presence of these myocotoxins
together reduces the toxic effects of the
mycotoxins separately (growth and feed
consumption decrease).
 BIAS OF THE ANALYTICAL
METHOD
• 1. Not representative sampling
Sample not representative, because of the big sample
size or the king of storage/container of the raw material.
2. Not validated analysis method
Analysis method should have been validated by a
prestigious institution like International Organization for
Standardization (ISO).
3. Low quality standards
Trouble to get mycotoxin standards in some countries.
% recovery of solid standards by dissolution <100%.
Standard solution not stable.
4. Procedure for sample extraction
PRESENCE OF NON-ANALYSED
MYCOTOXINS
• Not-analysed mycotoxins
In raw materials there could be mycotoxins whose are
not analysed:
1. WELL-KNOWN MYCOTOXINS Whose analysis is not
performed because of economic reasons, lack of
validated methods, presence unlikely, etc.
2. LITTLE-KNOWN MYCOTOXINS Mycotoxins whose
incidence and effects are little known.
THERE ARE MORE THAN 300 DESCRIBED
MYCOTOXINS
 Reference:
• Interpretation of the results of
mycotoxin's analysis in feed
• Author: Paper Presented at Biovet
Symposium 2007 (Courtesy of Biovet SA)