ANTI-NUTRITIONAL FACTORS

1. INTRODUCTION
Anti-nutritional substance are defined as those generated in feed stuffs by the normal
metabolism of the species from which the material originates and by different mechanism
exerting effects contrary to optimum nutrition. These anti-nutritive substances are often
referred to as toxic factors because of the deleterious effects they produce when eaten by
animals. The term toxic factor is misleading because there is an implication that the
substances are lethal beyond the certain level .
Anti-nutrients(Anti-nutritional factors)
Definition:
Substances which either by themselves or through their metabolic products, interfere with
food utilisation and affect the health and production of animals
General characteristics
 Products of secondary metabolism
 Found in virtually all plants to some degree
 Common in tropical forages
 Defensive role: Bitter, poisonous, bad odour, anti nutritive / immune suppresive
There are many anti nutritional factors present in feeds and fodders which
affect the utilization of nutrients .Some of these are toxic if they are consumed in large
quantities. Few of them are dealt here.

2. LITERATURE REVIEW
Despite the efficiency of thermal treatment to reduce protease inhibitors, residual
inhibition (10-20%) is maintained (Carvalho et al., 1998). Phytic acid (Inositol
hexaphosphoric acid) forms insoluble salts with essential minerals like calcium, iron,
magnesium and zinc in food rendering them unavailable for absorption into the blood stream
(Bingham, 1978). Phytic acid and its hydrolysis products are associated with inhibition of
calcification in rats (Robert andYudkin, 1999). The earliest interest in the toxicity of oxalate
arose because of instances of severe or fatal human poisoning following the eating of large
quantity of the leaves of certain plants i.e., rhubarb, known to contain relatively large
amounts of oxalates, (Osagie, 1998). Plants evolved these substances to protect themselves
and to prevent them from being eaten. However, if the diet is not varied, some of these
toxins build up in the body to harmful levels (Norman and Potter, 1987). Aflatoxin in
groundnut has been found to cause severe liver damage if eaten, they are heat stable and can
be degraded by strong acid and alkaline treatment (Harold and Pattee, 1985).
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3. CLASSIFICATION OF ANTINUTRITIONAL FACTORS
On the basis of the types of nutrients affected and the biological response produced in
the animal the toxic factors can be classified into 5 major groups as follows.
1. Substance depressing digestion or metabolic utilization of proteins
A. Protease inhibitors
B. Lectins of ricins (Haemagglutinins)
C. Saponins
D. Polyphenolic compounds (tannins)
2. Substaces reducing the solubility or interfering with the utilization of mineral
elements
A. Phytic acid
B. Oxalic acid
C. Glucosinolates
D. Gossypol
3. Substance inactivating or increasing the requirements of certain vitamins and
hormones
A. Antivitamins A, D, E, K and antipyridoxins
B. Mimosine antihormone
4. Cyanogens
5. Nitrate and nitrite
6. Moulds and mycotoxins in animal feedstuffs

1. Substance depressing digestion or metabolic utilization of proteins:

A. Protease inhibitor
Substances that inhibit proteolytic enzymes and thereby growth of nonruminants are
disturbed. In case of soybean two types of protease inhibitors are found
1. Kunitz inhibitors have few disulphide bonds and inhibit trypsin
2. Bowmanbrik inhibitors have a high proportion of disulphide bonds and inhibit trypsin
and chymotrypsin Feeding raw soybeans to pigs chicks and rats have resulted in
reduced growth rate , pancreatic hyperplasia and low production although ruminants
are capable of utilizing raw soybeans without suffering any deleterious effects, a
better response in milk production and growth rate is obtained on diets containing
treated soybean.

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 The inhibitory substances are mostly heat labile and thus before feeding any
leguminous grains to non ruminants, they must be treated by heat . Among the anti-nutritional
factors present in soybean seed, the main ones are protease inhibitors – Kunitz trypsin
inhibitor (KTI) and Bowman-Birk inhibitor, and lectins. Protease inhibitors represent 6% of
the protein present in soybean seed. Approximately, 80% of the trypsin inhibition is caused
by KTI, which strongly inhibits trypsin and therefore reduces food intake by diminishing
their digestion and absorption. Another effect of KTI is the induction of pancreatic enzyme,
hyper secretion and the fast stimulation of pancreas growth, hypertrophy and hyperplasia.
Due to this, raw soybean cannot be used for feeding monogastric animals. Heat treatment
doesn’t completely eliminate these factors and may decrease protein solubility. Despite the
efficiency of thermal treatment to reduce protease inhibitors, residual inhibition (10-20%) is
maintained (Carvalho et al., 1998). By this reason, a part of the breeding program of the
Maize Research Institute Zemun Polje is aimed at developing soybean cultivars with reduced
trypsin inhibitors content.

B. Lectins of ricins
It is found in both plant and animal tissue. In castor bean cakes a toxic fraction
capable of agglutinating human red blood cells was noted as ricins. Lectins are protein in
nature digestion resistant by pancreatic juice. Although very resistant to destruction by dry
heat, lectins are destroyed by the same conditions as those used to inactivate protease
inhibitors.
C.Saponins
The important common forages which have caused saponin poisoning in livestock are
lucerne, white clover, red clover and soybean.

Chemical structure and general properties

Saponins are plant glycosides which yield on hydrolysis sugars (pentose, hexose and
uronic acid) and aglycones derived from polycyclic ring systems and are known as
sapogenins. Saponins are divided into two main groups from the point of view of chemical
nature of sapogenins: steroids or triterpenoides with the triterpenoid sapogenins further
divided into three classes based on the compounds ursane and oleanane/lupine. They have
three significant characteristics
1. A bitter taste
2. Foaming in aqueous solution
3. Haemolysis of red blood cells
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 Due to bitter taste of saponins it has been suggested that the reduced feed intake is due
to unpalatability.
Biological activity of saponins can be related to the following characteristic properties
i. Structurally saponins have a lipid soluble aglycone and a water soluble glycine part
which jointly confers lower surface tension and thus forms stable foam when
dissolved in water/
ii. Most of the saponins readily combine with cholesterol resulting minimisation of the
activity of original saponins.
iii. They ossess the ability to haemolyse red blood cells
iv. They tend to alter the permeability of the cell wall and thereby exert a general
toxicity on many organized tissues
Adverse action on excessive eating
1. It increases the respiratory rate which later becomes irregular.
2. It inhibits action of certain enzymes for e.g. α-chymotrypsin.
3. In general the effects of injestion of saponins include excessive salvation, increase
respiratory tract secretion, gastroenteritis, vomiting, diarrhoea, haemolysis,
haematuria and damage to livers and kidney tissues, cystitis, bloating, reduction in
gastric motility, reduction of cholesterol absorption from the gut, lowering of blood
and liver cholesterol levels, reduction of food intake and growth rate.

C. Polyphenolic compounds (tannins)

It is present in number of vegetable extracts which are responsible for converting
putrescible animal skin into the stable product leather by the tanning process. It is also known
as tannic acid, gallotannin, gallotannic acid. These substances are also termed as group of
phenolic non-nitrogenous plant toxins that are frequently glycosides with astringent
properties.
Types of tannin
Chemically tannins may be grouped into two main categories
1. Hydrolysable tannins
2. Condensed tannins
Most tannin abstracts appear to contain mixture of both types of tannins but generally
one or the other predominates at least in any given part of the plant

1. Hydrolysable tannins

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 It can be readily hydrolysed by water, acids, bases or enzymes. Hydrolysable tannins
are those in which gallic acid and its related compounds like hexahydro-xydiphenic acid are
linked in sufficient proportion to a sugar by glycosidic linkages to provide polyphenolic
compounds of relatively high molecular weight. Such compounds are also know as
gallotannins because of the presence of gallic acid. On hydrolysis such compounds yield
glucose or some other polyhydric alcohol together with gallic acid or other phenolic acids
related to it.

2. Condensed tannin
Compounds contain only phenolic nuclei although polysaccharides or proteins may be
irreversibly linked to them in vivo or during the course of isolation. On treatment with
hydrolytic reagents tannins of this class give no significant yields of lower molecular weight
compounds but instead tend to polymerise especially in acid solution to yield insoluble
amorphous red colured products known as phlobaphene. Most tannis of this type are formed
by the condensation of two or more molecules of flavan-3-ons such as catechin or flavin-3,4-
diols such as leucocyanidin or the mixture of the two. Tannins of this type are usually termed
non-hydrolysable or more commonly condensed.since the majority of tannins appear to be
formed by the polymerization of flavans, it is convenient to refer to this group specifically as
flavolans and to use the term condensed tannin in this wider sense.

Distribution of tannins in livestock feeds and fodders

Name of the feed stuff Approx. %
Sorghum 0.004-10.50
Milo 2.00-3.00
Salseed meal 9.00-12.00
Mango seed kernel 5.00-7.00
Mustard oil cake 2.80-3.20
Rape seed cake 3.00-3.50
Lucerne meal 0.10-3.50
Methods of detannification
1. Physical treatments:

 Soaking and cooking of tannin containing feed stuffs have been found very effective
in decreasing the tannin content. However, these treatments cause a substantial loss of
dry matter between 20-70 %.

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  Anaerobic storage of moist sorghum grains for two and nine days at 25 0C resulted in
40% and 90% reduction in tannins respectively. The nutritive value of the treated
grains was found to be higher.

2.Chemical treatments:

a) Addition of tannin complexing agents like polyethylene glycol (PEG) and

polyvinyl pyroldone(PVP) prevent formation of complexes between tannin and
protein as well as break the already formed complexes, thus liberating protein.

b) Treatment with alkalies like NaOH, Ca(OH) 2 and lime water of sal seed meal was
found to be very effective in removing tannins(74-100%), but treatments with Na2CO3
and NaHCO3 were comparatively less effective( about 50%) removal.

c) Formaldehyde treatment
d) Treatment with methanol and acetone
e)Treatment with 0.1% HCl

2.Substances reducing the solubility or interfering with the utilization of mineral

elements
Gossypol
It is a toxic phenolic compound present in cotton seeds. Heat treatment destroys the
gossypol. In ruminants ,it is less toxic.
Glucosinolates:
These are found in almost all the species of crucifera family. The plants , seeds and oil
cakes of different mustard and rape varieties are rich sources of this glucosinolates .These
compounds reduce the incorporation of iodine into the precursor of thyroxine resulting in
iodine deficiency and development of goiter. Prolonged water soaking or cooking of feeds
inactivates the effects of goiterogens

Phytic acid:
Phytic acid (Inositol hexaphosphoric acid) forms insoluble salts with essential
minerals like calcium, iron, magnesium and zinc in food rendering them unavailable for
absorption into the blood stream (Bingham, 1978). About half of the phytic acid phosphorus

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content taken by man is excreted unchanged. The remaining unavailable for utilization, the
knowledge of phytate content can be made use of in calculating the quantity of phosphorus
available out of the total phosphorus in a diet. Phytic acid and its hydrolysis products are
associated with inhibition of calcification in rats (Robert andYudkin, 1999).
Oxalate:
The name oxalate is attributed to its occurrence in plant oxalis (Wood sorel). The
earliest interest in the toxicity of oxalate arose because of instances of severe or fatal human
poisoning following the eating of large quantity of the leaves of certain plants i.e., rhubarb,
known to contain relatively large amounts of oxalates, (Osagie, 1998).

3.Substance inactivating or increasing the requirements of certain vitamins:

a) Anti-Vitamin A:
Raw soyabeans contain an enzyme Lipoxygenase, which catalyses oxidation of
carotene, the precursor of vitamin-A. It has been noted that 30% of ground, raw soyabeans in
the diet of dairy cattle produces a sharp lowering of vitamin A and carotene in blood plasma.
The enzyme can be destroyed by heating soyabeans for 15 minutes with steam at atmospheric
pressure.

b) Anti-Vitamin D:
Rachitogenic activity of isolated soya protein(unheated) has been found with chicks
and pigs. The effect could be partially eliminated by increasing the vitamin D in diet by 8-10
folds. Autoclaving eliminates the rachitogenic activity.

c) Anti-Vitamin E:

containing raw kidney beans (phaselous vulgaris) produce muscular dystrophy in

lambs by reducing plasma vitamin E. Alcohol extraction of the beans reveals 2 factors with
anti-vitamin E activity, one being alcohol soluble heat stable, the other being heat liable and
alcohol insoluble. By autoclaving beans the anti vitamin activity is eliminated.

d) Anti-Vitamin K:
“Sweet clover disease” is characterised by a fatal haemorrhagic condition in cattle and
has been known over 20 years. The active principle responsible for this disease is dicoumarol,
which reduces the prothrombin level of the blood, thus interfering with blood clotting

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mechanisms. The effect is due to reducing vitamin K utilisation in the production of thrombin
by the liver
e) Anti-Pyridoxine:

It has been demonstrated that the nutritive value of linseed meal for chicks can be
considerably improved after extracting the meal with water and autoclaving. An antagonist of
pyridoxine, (a member of B vitamins) from linseed which has been identified as 1- amino-D-
proline, and occurs naturally in combination with glutamic acid as a peptide is known as
linatine.

f) Mimosine (Leucaena leucocephala):

The plant subabul and other legumes contains a toxic substance known as Mimosine
which is free amino acid, present at about 3 to 5% of dry matter. The young leaves are
relatively richer in mimosine content.
Mimosine , is also present in leucocephala forage and its seeds. The poor
nutritive value of leucocephala arises principally from its toxic amino acid, mimosine and its
immediate degradation product, i.e., 3-hydroooxy-4(1H) pyridine(DHP). The toxicity
depends upon different classes of ruminants and on the different geographical regions of the
same class of ruminants. Alopecia, loss of appetite , excessive salivation, in coordination of
gait, enlarged thyroid gland and poor breeding performance were reported in cattle and
buffaloes grazing/fed on lucaena. Poor wool growth and haemorrhagic cystitis were observed
in sheep fed on leucaenia diet .Thyroid enlargement was also observed. In goats also alopecia
and enlarged thyroid were observed. In some geographical regions leucaena feeding didnot
show any toxicity symptoms in ruminants like in Hawaii.
The Mimosine acts as an analog of tyrosine and blocks its iodination for the synthesis
of hormone thyroxine by inhibiting the enzymes tyrosinase and tyrosine carboxylase. The
mimosine occurs in almost all parts of the plants and its level in young plants is about 12
percent of dry matter and 3 to 5 percent in the seeds.
Mimosine is degraded in the rumen by the enzymes mimosinase produced by bacteria
to 3,4 DPH which is further broken down to pyruvic acid and ammonia. These bacteria are
present in ruminants

4.Cyanogens:

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 In plants the glucoside is non-toxic in the intact issues and as stated earlier, when the
plants are damaged or begin to decay, hydrolytic enzymes from the same plant is released,
liberating HCN. This reaction can take place in the rumen by microbial activity. The HCN is
rapidly absorbed and some is eliminated through the lungs, but the greater part is rapidly
detoxified in the liver by conversion to thiocyanate. Excess cyanide ion can quickly produce
anoxia of the central nervous system through inactivating the cytochrome oxidase system,
and death can result within a few seconds. Based on the intensity animal shows nervousness,
abnormal breathing, trembling or jerking muscles, blue coloration of the lining of the mouth,
spasms or convulsions and respiratory failure.

5.Nitrates and Nitrites:

Animal forages and drinking water when contaminated with inorganic nitrates and
nitrites causes an acute toxicosis in cattle resulting from the formation of methmoglobin(a
true oxidation product of haemoglobin) which is unable to transport oxygen because the iron
is in the ferric form rather than the usual ferrous state. Symptoms seen in acute toxicity
include laboured breathing (dyspena), grinding of teeth, uneasiness and excessive salivation.

6.Aflatoxins:
Aflatoxins are common term used for a group of toxins used for a group of toxins
which differ in their chemical structure and in intensity of producing toxic effects. As the
various compounds of this group are produced by the fungus Aspergillus flavus and A.
Parasiticus they resemble very much to each other and are commonly termed aflatoxins.
The production of toxins by the above two fungi depends on the following
conditions:
1. The strain of the fungus
2. The nature of the substrate
3. The temperature and humidity of the environment immediately surrounding the
area of mould growth.
4. The optimum temperature of the aflatoxins is around 270C.

Toxicosis in farm animals by aflatoxins:

Species Age Aflatoxin content effects
(mg/kg;ppm)

Calf Weaning 0.2-2.2 Stunting, death, liver

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 damage
Steer 2 yrs 0.2-0.7 Liver damage
Cow 2 yrs 2.4 Liver damage
Pig Newborn 0.23 Stunting
Pig 2 weeks 0.17 Anorexia, Stunting
Pig 4-6 weeks 0.4-0.7 Stunting, Liver
damage
Chicken 1+ weeks 0.8 Stunting, Liver
damage
Duck 0.3 Liver damage, death

4. CONCLUSION:
Anti-nutritional factors are present in different food substances in varying amounts,
depending on the kind of food, mode of its propagation, chemicals used in growing the crop
as well as those chemicals used in storage and preservation of the food substances It is found
in most food substances which are poisonous to animals or in some ways limit the nutrient
availability to the body.
Plants evolved these substances to protect themselves and to prevent them from being eaten.
However, if the diet is not varied, some of these toxins build up in the body to harmful levels
(Norman and Potter, 1987). Some vitamins in food may be destroyed by anti-nutritional
substances. Aflatoxin in groundnut has been found to cause severe liver damage if eaten, they
are heat stable and can be degraded by strong acid and alkaline treatment (Harold and Pattee,
1985). These anti-nutritional factors must be inactivated or removed, if values of food
substances are to be fully maintained.

BIBLIOGRAPHY :
 Banerjee G.C.1998. Feeds and principles of Animal Nutrition. Oxford & IBH
publishing Co. PVT. LTD.
 McDonald, P., R.A.Edwards and IFD Greenhalgh. 1987. Animal Nutrition ELBS
Publication(4th Edition)
 Morrison, F.B.1984.Feeds and Feeding. C.B.S Publishers and Distributors, Jain
Bhawan, New Delhi, India
 Ranjhan S.K.1993 Animal Nutrition and Feeding Practices in India. Vikash
Publishing House Pvt. Ltd., India

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