Tannins (commonly referred to as tannic acid) are water-soluble polyphenols that are present in many

plant foods. They have been reported to be responsible for decreases in feed intake, growth rate, feed
efficiency, net metabolizable energy, and protein digestibility in experimental animals. Therefore, foods
rich in tannins are considered to be of low nutritional value. However, recent findings indicate that the
major effect of tannins was not due to their inhibition on food consumption or digestion but rather the
decreased efficiency in converting the absorbed nutrients to new body substances. Incidences of certain
cancers, such as esophageal cancer, have been reported to be related to consumption of tannins-rich
foods such as betel nuts and herbal teas, suggesting that tannins might be carcinogenic. However, other
reports indicated that the carcinogenic activity of tannins might be related to components associated with
tannins rather than tannins themselves. Interestingly, many reports indicated negative association
between tea consumption and incidences of cancers. Tea polyphenols and many tannin components
were suggested to be anticarcinogenic. Many tannin molecules have also been shown to reduce the
mutagenic activity of a number of mutagens. Many carcinogens and/or mutagens produce oxygen-free
radicals for interaction with cellular macromolecules. The anticarcinogenic and antimutagenic potentials of
tannins may be related to their antioxidative property, which is important in protecting cellular oxidative
damage, including lipid peroxidation. The generation of superoxide radicals was reported to be inhibited
by tannins and related compounds. The antimicrobial activities of tannins are well documented. The
growth of many fungi, yeasts, bacteria, and viruses was inhibited by tannins. We have also found that
tannic acid and propyl gallate, but not gallic acid, were inhibitory to foodborne bacteria, aquatic bacteria,
and off-flavor-producing microorganisms. Their antimicrobial properties seemed to be associated with the
hydrolysis of ester linkage between gallic acid and polyols hydrolyzed after ripening of many edible fruits.
Tannins in these fruits thus serve as a natural defense mechanism against microbial infections. The
antimicrobial property of tannic acid can also be used in food processing to increase the shelf-life of
certain foods, such as catfish fillets. Tannins have also been reported to exert other physiological effects,
such as to accelerate blood clotting, reduce blood pressure, decrease the serum lipid level, produce liver
necrosis, and modulate immunoresponses. The dosage and kind of tannins are critical to these effects.
The aim of this review is to summarize and analyze the vast and sometimes conflicting literature on
tannins and to provide as accurately as possible the needed information for assessment of the overall
effects of tannins on human health.

Tannins in Tea
Wikipedia: Tannin
Last Updated: Mar. 20, 2014
by Labrau, licensed under CC BY-SA 3.0.

Tannins are a class of compounds in tea, especially black tea, which tend to
have a bitter flavor and astringent properties. Teas high in tannins can be
described as tannic.
Tannins are naturally occurring and common, and are an important
component of red wine. They are also responsible for the dark color in some
streams, as they are found in leaves and wood, and are released as organic
matter breaks down.
The name "tannin" originated in the historical use of these compounds in

tanning animal hides to make leather.

Tasting and seeing tannins

People talk of some teas having a "tannic quality", which usually describes the
pairing of a sharp bitterness with a strong astringency, in black teas with a
dark color. This bitterness contrasts with the bitterness imparted by caffeine,
which need not be paired with astringency, and can occur in teas with a very
light color.
Less oxidized teas, like green tea, can be highly astringent and bitter even
while being light in color. This is in large part because they contain other
chemicals, such as catechins, which are share certain similarities with tannins.

Myths about tannins and health

Some common myths and misconceptions about the tannins in tea:

Tea contains tannic acid - False. Tea does not contain any tannic acid.
This myth stems from confusion between tannins (a broad class of
compounds) and tannic acid, a specific type of tannin. Tea contains
tannins other than tannic acid.
Tannins are bad for you. - False. As explained below, tannins have both
positive and negative effects on health, and like most substances, are
healthiest in moderation.
Tannins taste bad or make a tea low quality - False. Too high a
concentration of tannins in a cup of tea can result in an unpleasantly
astringent cup, but tannins also impart a sensation of body and strength
to a tea, and make for a richer mouthfeel: a black tea with too few
tannins would be more likely to seem watery, weak, or bland.

Tannins are responsible for the rich reddish-brown colors in some rivers and streams. Public domain photo byPseudopanax.

American society, unfortunately, has had numerous health fads characterizing

whole class of compounds as either "good" or "bad", and tannins have fallen
onto the "bad" side of public perception, much as antioxidants have fallen onto
the "good" side. Most people, however, are surprised to learn that the tannins
in tea are antioxidants. Our page on antioxidants explains more.
James Norwood Pratt in his Tea Dictionary goes even farther, claiming that
the term "tannins" is an obsolete term for the polyphenols in tea. He notes that
the term was adopted in the 1800's, when the distinction between the
polyphenols and tea and the tannic acids in oak were not well-understood.[1]
However, the term tannin is widely used and is an accepted scientific term;
although the association between tannic acid and tea has long been viewed
as a misconception[2], the broader class of compounds, including the
polyphenols in black tea, are commonly described as tannins, even in the
contemporary scientific literature.

What types of tannins are found in tea?

The tannins found in tea are called thearubigins, a class of chemicals which
includes theaflavins. These chemicals are formed in black tea when the
antioxidants inherent in green tea, called catechins, becomeoxidized. Tea does
not contain any tannic acid.[2]
The tannins in tea are thus responsible for the antioxidant activities of black
tea and other dark (oxidized) teas, including more-oxidized oolong teas.
However, these same chemicals can also have negative impacts on health;
the tannins are also responsible for tea's inhibiting effect on iron absorption.[3]

Not all dark-colored substances in tea are tannins

Many teas, including some oolongs, or Japanese hojicha, are roasted, giving
them a dark color which superficially resembles black tea. These other
substances have a distinct flavor and mouthfeel--as is evidenced by the much
smoother flavor and lower astringency of hojicha and some roasted oolongs,
relative to black teas that are similarly dark. Oolongs that are both oxidized
and roasted can have a dark color from a combination of tannins and various
compounds formed by the roasting.

Nutritional and health effects of tannins

Tannins are a diverse class of compounds, and have diverse effects on
health. Tannins are often consideredantinutritional, as animal studies suggest
they can reduce net metabolizable energy and protein digestibility. However,
these same compounds often exhibit anti-carcinogenic and anti-mutagenic
properties, probably due to their antioxidant activity. Tannins also have welldocumented antimicrobial properties, effective against many bacteria, fungi,
and viruses.[4]
Many specific compounds within the class of tannins have other biological
activity or medical uses, which include speeding of blood clotting, reduction of
blood pressure, and effects on the liver and immune system.[4]

Low tannin teas (especially black teas)

People who dislike teas with tannic qualities, or wish to avoid tannins for
medical reasons, have a lot of options. Green and white teas, and lowoxidation oolongs tend to contain little to no tannins, although they are rich in
the chemically-similar catechins, sometimes classified as pseudotannins.
Some low-tannin black teas include:

Darjeeling first flush - Some Darjeeling first flush teas are unusually light in

character; this is due in large part to the thin, dry air that exists in the
early spring at high altitudes. First flush Darjeeling teas are harvested
from the new growth that follows just after the first spring rains, so the
air is still thinner and drier. This makes the leaf dry out faster during
processing, so it does not oxidize fully, thus retaining similar
characteristics to green teas or greener oolongs.

Gold teas, including Yunnan pure gold and golden monkey - Some highgrade black teas are made mostly or exclusively from golden tip. These
teas can have a very light color to the dry leaf, and, although they
produce a dark-colored infusion when brewed, are often markedly lower
in tannins than their lower-grade counterparts. Be careful though, as
many teas labelled as "Yunnan Gold" or "gold tip" teas still contain a lot
of larger, more mature leaves, and can still be quite tannic.
References
1. James Norwood Pratt, Tea Dictionary, 2010, pp. 263.
2. Stephen R. Wheeler, Tea and Tannins, Science, Letters, Vol. 204, No. 4388, pp. 6-8 , April 1979.
3. Effect of tea and other dietary factors on iron absorption., Critical Reviews in Food Science and
Nutrition, Sep. 2000, Vol. 40, No. 5, pp. 371-98.
4. Chung KT, Wong TY, Wei CI, Huang YW, Lin Y., Tannins and Human Health: A Review, Critical
Reviews in Food Science and Nutrition, Vol. 38, No. 6, pp. 421-64, Aug. 1998.

Tannins: fascinating but sometimes dangerous

molecules
Welcome to the Tannin webpage. We offer a variety of information on tannins
including, but not limited to, their biosynthesis, chemical structures, toxicology,
positive effects, chemical analysis....
Tannins are naturally occurring plant polyphenols. Their main characteristic is that
they bind and precipitate proteins. They can have a large influence on the
nutritive value of many foods eaten by humans and feedstuff eaten by animals.
Tannins are common in fruits (grapes, persimmon, blueberry, etc.), in tea, in
chocolate, in legume forages (trefoil, etc.), in legume trees
(Acacia spp., Sesbania spp., etc.), in grasses (sorghum, corn, etc.).
Tannins contribute to many aspects of our daily lives. They are responsible
for the astringent taste we experience when we partake of wine or unripe
fruits, and for the enchanting colors seen in flowers and in autumn leaves.
For more information on tannins explore the following topics:
[Definition] [Occurrence] [Biosynthesis] [Chemical
structure] [Interaction with other macromolecules] [Nutritional
effects] [Chemical analysis] [References] [Return to list of toxicants]
Definition
The word tannin is very old and reflects a traditional technology. "Tanning" (waterproofing
and preserving) was the word used to describe the process of transforming animal hides
into leather by using plant extracts from different plant parts of different plant species.

o
o

Plant parts containing tannins include bark, wood, fruit, fruitpods, leaves, roots, and
plant galls.
Examples of plant species used to obtain tannins for tanning purposes are wattle
(Acacia sp.), oak (Quercus sp.), eucalyptus (Eucalyptus sp.), birch (Betula sp.), willow
(Salix caprea), pine (Pinus sp.), quebracho (Scinopsis balansae) .

Tannins are phenolic compounds that precipitate proteins. They are composed of a very diverse group of
oligomers and polymers. There is some confusion about the terminology used to identify or classify a substance as a
tannin, In fact,

o
o

not only tannins bind and precipitate proteins (other phenolics such as pyrogallol and resorcinol also have this
property),
not all polyphenols precipitate proteins or complex with polysaccharides.

One of the most satisfactory definition of tannins was given by Horvath (1981):
"Any phenolic compound of sufficiently high molecular weight containing sufficient hydroxyls and other suitable
groups (i.e. carboxyls) to form effectively strong complexes with protein and other macromolecules under the
particular environmental conditions being studied"
Tannins can complex with:

Proteins
Starch
Cellulose
Minerals

Occurrence
Tannins are widely distributed in the plant kingdom. They are common
both in Gymnosperms and Angiosperms. Within Angiosperms, tannins
are more common in Dicotyledons than in Monocotyledons.
Examples of families of Dicotyledons rich in tannins are:

Leguminosae : Acacia sp. (wattle); Sesbania sp.; Lotus sp.

(trefoil); Onobrychis sp. (sainfoin);
Anacardiaceae: Scinopsis balansae (quebracho)
Combretaceae: myrobalan
Rhizophoraceae : mangrove
Myrtaceae: Eucalyptus sp., Mirtus sp. (Myrtle)
Polinaceae: canaigre.

Other important tannin containing plants are Quercus sp. (oak), Acer sp. (maple), Betula sp. (birch), Salix
caprea (willow), Pinus sp. (Pine), Sorghum sp.
Tannins are located mainly in the vacuoles or surface wax of the plants. In these sites they do not interfere with
plant metabolism. Only after cell breakdown and death can they act and have metabolic effects.
Location of the tannins in various plant tissues:

Bud tissues - most common in the outer part of the bud, probably as protection against freezing
Leaf tissues - most common in the upper epidermis. However, in evergreen plants, tannins are evenly
distributed in all leaf tissues. They serve to reduce palatability and, thus, protect against predators.
Root tissues - most common in the hypodermis (just below the suberized epidermis). They probably act as a
chemical barrier to penetration and colonization of roots by plant pathogens.
Seed tissues - located mainly in a layer between the outer integument and the aleurone layer. They have
been associated with the maintainance of plant dormancy, and have allelopathic and bactericidal properties.
Stem tissues - often found in the active growth areas of the trees, such as the secondary phloem and xylem
and the layer between epidermis and cortex. Tannins may have a role in the growth regulation of these
tissues. They are also found in the heartwood of conifers and may be a contribute to the natural durability
of wood by inhibiting microbial activity.

Biosynthesis

Shikimic acid pathway.

There are three large classes of secondary metabolites in plants:
1.
2.
3.

Nitrogen containing compounds

Terpenoids
Phenolics

Tannins belong to the phenolics class. All phenolic compounds (primary and secondary) are, in one way or another,
formed via the shikimic acid pathway, also known as the phenylpropanoid pathway.
The same pathway leads to the formation of other phenolics such as isoflavones, coumarins, lignins and aromatic
aminoacids (tryptophan, phenylalanine and tyrosine).
The two main categories of tannins that impact animal nutrition are hydrolyzable tannins (Hts) and condensed tannins
identified more correctly as proanthocyyanidins (Pas) that are resistant to hydrolytic degragation. An example of how
several common tannins are formed is as follows:

o
o
o
o

Gallic acid is derived from quinic acid.

Ellagotannins are formed from hexahydroxydiphenic acid esters by the oxidative coupling of neighboring
gallic acid units attached to a D-glucose core.
Further oxidative coupling forms the hydrolyzable tannin (HT) polymers.
Proanthocyanidin (PA) biosynthetic precursors are the leucocyanidins (flavan-3,4-diol and flavan-4-ol)
o Upon autoxidation, in the absence of heat, they form anthocyanidin and 3-deoxyanthocianidin,
which, in turn, polymerize to form PAs.

Chemical Structure
Tannins are one of the many types of secondary compounds found in plants
Characteristics of tannins:

oligomeric compounds with multiple structure units with free phenolic groups,
molecular weight ranging from 500 to >20,000,
soluble in water, with exception of some high molecular weight structures,
ability to bind proteins and form insoluble or soluble tannin-protein complexes.

Tannins are usually subdivided into two groups:

Hydrolyzable tannins (HT)

Proanthocyanidins (PA) (often called Condensed Tannins)

Hydrolyzable tannins

HTs are molecules with a polyol (generally D-glucose) as a

central core. The hydroxyl groups of these carbohydrates are
partially or totally esterified with phenolic groups like gallic acid
(-->gallotannins) or ellagic acid (--> ellagitannins). HT are
usually present in low amounts in plants.
Some authors define two additional classes of hydrolyzable
tannins: taragallotannins(gallic acid and quinic acid as the
core) and caffetannins (caffeic acid and quinic acid)
1. Gallotannins:

The phenolic groups that esterify with the core are

sometimes constituted by dimers or higher oligomers
of gallic acid (each single monomer is called galloyl)
Each HT molecule is usually composed of a core of D-glucose and 6 to 9 galloyl groups
In nature, there is abundance of mono and di-galloyl esters of glucose (MW about 900). They are not
considered to be tannins. At least 3 hydroxyl groups of the glucose must be esterified to exhibit a sufficiently
strong binding capacity to be classified as a tannin.
The most famous source of gallotannins is tannic acid obtained from the twig galls of Rhus semialata. It has
a penta galloyl-D-glucose core and five more units of galloyl linked to one of the galloyl of the core.

2. Ellagitannins:

The phenolic groups consist of hexahydroxydiphenic acid, which spontaneously dehydrates to the lactone
form, ellagic acid.
Molecular weight range: 2000-5000.

HT properties:

hydrolyzed by mild acids or mild bases to yield carbohydrate and phenolic acids
Under the same conditions, proanthocyanidins (condensed tannins) do not hydrolyze.
HTs are also hydrolyzed by hot water or enzymes (i.e. tannase).

Proanthocyanidins (condensed tannins)

PAs are more widely distributed than HTs. They are oligomers or polymers of
flavonoid units (i.e. flavan-3-ol) linked by carbon-carbon bonds not
susceptible to cleavage by hydrolysis.

PAs are more often called condensed tannins due to their condensed
chemical structure. However, HTs also undergo condensation reaction. The
term, condensed tannins, is therefore potentially confusing.

The term, proanthocyanidins, is derived from the acid catalyzed oxidation reaction that produces red anthocyanidins
upon heating PAs in acidic alcohol solutions.

The most common anthocyanidins produced are cyanidin (flavan-3-ol, from procyanidin) and delphinidin
(from prodelphinidin)

PAs may contain from 2 to 50 or greater flavonoid units; PA polymers have complex structures because the flavonoid
units can differ for some substituents and because of the variable sites for interflavan bonds.

Anthocyanidin pigments are responsible for the wide array of pink, scarlet, red, mauve, violet, and blue colors in
flowers, leaves, fruits, fruit juices, and wines. They are also responsible for the astringent taste of fruit and wines.

PA carbon-carbon bonds are not cleaved by hydrolysis.

Depending on their chemical structure and degree of polymerization, PAs may or may not be soluble in aqueous
organic solvents.
Interaction with other Macromolecules
Tannins have a major impact on animal nutrition because of their ability to form complexes with numerous types of
molecules, including, but not limited to,

Carbohydrates,
Proteins,
Polysaccharides,
Bacterial cell membranes,
Enzymes involved in protein and carbohydrates digestion.

Carbohydrates
Both starch and cellulose are complexed by tannins (especially by PAs):

Starch-tannin interaction - starch has the ability to form hydrophobic cavities that allow inclusion complexes with
tannins and many other lipophyllic molecules. Only starch, among the molecules that are bound by tannins, has this
embedding characteristic.
Cellulose-tannin interaction - cellulose has a direct surface interaction with tannins.
Cell wall carbohydrate-tannin interaction - this association is less understood. One explanation is that tannins
associate with plant cell walls in a manner reminiscent to that of lignin. However, another explanation is that this
association is merely an artifact of tannin isolation from non-living cells. Indeed, the location of tannins and cell wall
carbohydrates is quite different in living cells than in plant cells after digestion by animals.
Tannin-carbohydrate interactions are increased by carbohydrates with high molecular weight, low solubility and
conformational flexibility. These interactions are probably based on hydrophobic and hydrogen linkages.

Proteins
The capacity of tannins to bind proteins has been recognized for centuries. Leather tanning is a very ancient practice.
Tannin-protein interactions are specific and depend on the structure of both the protein and tannin.

Protein characteristics that favor strong bonding

large molecular size,
open and flexible structures,
richness in proline.
Tannin characteristics that favor strong bonding
high molecular weight,
high conformational mobility.

Chemical linkages
Tannin-protein interactions are most frequently based on hydrophobic and hydrogen bonding. Ionic and covalent
bonding occur less frequently.

The tannin's phenolic group is an excellent hydrogen donor that forms strong hydrogen bonds with the protein's
carboxyl group.

For this reason, tannins have a greater affinity to proteins than to starch.

Hydrophobic bonds are stronger at higher ionic strength (higher tannin/protein ratios) and higher temperatures.

Covalent bonding occurs only under oxidizing conditions such as

autoxidation over time, or

action of oxidative enzymes (i.e. polyphenoloxydases and peroxidases).

Covalent bonding is far more difficult to disrupt than the previous types of bonding and is nutritionally very
important because of its irreversible nature.

Precipitation of proteins by tannins is maximum at pH values near the isoelectric point of the protein.

In solution at high pH, phenolic hydroxyls are ionized and proteins have net negative charges. Under these
conditions, precipitation does not occur because proteins exhibit repulsive forces.

Strong complexes with tannins are formed by tannin-binding agents like polyvinylpyrrolidone (PVP) and polyethylen
glycol (PEG), and protein denaturants like phenol.

To have high protein affinity, tannins must be small enough to penetrate interfibrillar region of protein molecules but
large enough to crosslink peptide chains at more than one point.

HTs and PAs form tannin-protein complexes in similar manners. Proteins thus bound are generally resistant to attack
by proteases and hence may be unavailable for livestock nutrition. However, it is hypothesized that HTs may have a
less damaging effect on protein digestion because these tannins may hydrolyze in the acidic gastric environment and
release the bound proteins.

When soluble tannins interact with proteins, both soluble and insoluble complexes are formed; their relative
proportion depends on the concentration and size of both molecules.

Soluble complexes are favored when protein concentration is in excess (fewer tannin attachment sites per
each protein molecule). Soluble complexes represent an analytical problem because they do not precipitate
and, thus, are difficult to measure.

Insoluble complexes are formed when tannins are present in excess and form an hydrophobic outer layer
in the complex surface.

Nutritional Effects: toxic and antinutritional effects

Tannins act as a defense mechanism in plants against pathogens, herbivores and hostile environmental conditions.
Generally, tannins induce a negative response when consumed. These effects can be instantaneous
like astrigency or a bitter or unpleasant taste or can have a delayed response related to antinutritional/toxic effects.
This section will cover the effect of tannins on:
Intake, Feed digestibility, Toxicity to microorganisms, Toxicity to ruminants, Toxicity to
monogastric, Animals' defense mechanisms
Tannins negatively affect an animal's feed intake, feed
digestibility, and efficiency of production. These effects
vary depending on the content and type of tannin ingested
and on the animal's tolerance, which in turn is dependent
on characteristics such as type of digestive tract, feeding
behavior, body size, and detoxification mechanisms.
Sites of action of tannins :

Oral cavity - mastication ruptures the plant cell tissue and

exposes proteins and carbohydrates to tannins.

Rumen and gastrointestinal tract lumen - unbound tannins complex dietary proteins and metabolic proteins (e.g.
bacteria, enzymes, epithelial cells).

Intake
Tannins may reduce intake by decreasing palatability and by negatively affecting digestion.

Palatability is reduced because tannins are astringent. Astringency is the sensation caused by the formation of complexes
between tannins and salivary glycoproteins.

Digestibility reduction (see following section) negatively influences intake because of the filling effect associated with
undigested feedstuff.
Several studies have reported higher feed intakes and weight gains when tannin-free diets were compared to tannincontaining ones.

o
o
o
o

Low palatability depresses feed intake and, thus, animal productivity.

Some caution must be taken when interpreting these results. In many trials, commercial tannins sources were used. These types
of tannins are usually more effective at lowering feed intakes than naturally-occurring tannins.
Another likely problem in many trials is that often only extractable tannins are measured and insoluble tannins are not quantified.
However, insoluble tannins may have equal or greater biological activity than those that are more easily extracted.
When naturally-occurring tannins are used, these tannins do not always reduce intake. In some trials, tannin-rich diets were eaten
in equal or larger amounts than low or free tannin diets.

The form in which the forage is fed may influence how tannins affect feed intake. Forages rich in tannins are eaten in
larger amounts when field dried rather than fresh frozen. Indeed, drying reduces the solubility of tannins and, hence,
reduces their ability to complex proteins (tannins become more polimerized, resulting in a lower number of free hydroxyls
available for binding the proteins).
Intake in animal diets rich in tannins can be increased by using a compound with a high affinity for tannins, like PEG
(polyethylene glycol).

o
o
o

PEG has a higher affinity to tannins than do proteins.

PEG can be sprayed on the forages or added in the diet and is fairly inexpensive.
PEG utilization can increase feed palatability and digestibility and result in higher animal productivity.

Feed intake may also be decreased by low molecular weight phenolics. They predominate during the early stages of
plant growth and are then converted to oligomers and finally to polymers (tannins) when the plant mature. These low MW
phenolics are absorbed into the body and exhibit systemic effects such as alteration of physiological systems, increased
energy requirements due to detoxification, and subsequent growth rate reduction.

Feed digestibility
Usually PAs are not absorbed through the digestive tract. Instead, free tannins and complexed forms remain in the
rumen, decreasing protein and plant cell wall digestibility.
Carbohydrate digestibility

Several studies have shown that tannins decrease organic matter and fiber digestion.
The lower digestibility is the result of the interaction of tannins with cellulase enzymes and rumen bacteria.
In some cases, lower fiber digestibility can be the result of a shortage of ruminally fermented nitrogen due to the
complexation of proteins by tannins.
Field drying and treatments with PEG are able to limit these negative effects.
In some cases, lower digestibility was compensated by higher protein outflow from the rumen.
Protein digestibility

In in vivo studies, protein digestibility is greatly reduced when tanniniferous feeds are part of the diet.
Plants high in PAs often have proteins linked tightly to the plant cell wall (neutral-detergent insoluble nitrogen, NDIN)
and lignin (acid detergent lignin, ADL) components, and, thus, may show negative digestion coefficients when
ingested.
After ingestion, PAs may also form detergent insoluble tannin-protein complexes with proteins they
encounter. These two factors may cause the amount of NDIN and ADL excreted in the feces to exceedthe
amount ingested.
When dietary content of tannins increases, fecal nitrogen excretion increases due to lower digestibility of nitrogen
fractions and formation of tannin-protein complexes.

However, despite the decrease in apparent nitrogen digestibility, nitrogen retention does not always decrease with
increasing tannin concentration in the diet . In many cases, nitrogen retention increases as a result of
decreased urinary excretion.
The effect of tannins on protein digestibility of a specific feed can be estimated using Lucas's test (Van Soest, 1994,
page 360).
If protein digestibility is not affected by tannins, proteins behave as a uniform fraction, with a regression
coefficient (true digestibility) equal to or larger than 0.88, with a negative intercept(estimate of metabolic
endogenous nitrogen, usually about 0.5% of dry matter intake or smaller) and with a low standard error.
However, if protein digestibility is affected by tannins, the proteins will behave as a non-uniform fraction,
with a regression coefficient (true digestibility) smaller than 0.88, and with a larger negative intercept and a
higher standard error
Tannin solubility plays a role in determining a tannin's efficiency in binding proteins and/or fiber.
If the ratio of soluble to insoluble tannins is high, then protein digestibility is affected more than fiber
digestibility.
If the same ratio is low, fiber digestibility is the most affected.

Toxicity to microorganisms
Tannin toxicity to rumen microorganisms has been described for several bacteria species such as Streptococcus
bovis, Butyvibrio fibrosolvens, Fibrobacter succinogenes, Prevotella ruminicola, and Ruminobacter amylophilis.

Three mechanisms of toxicity have been identified

enzyme inhibition and substrate deprivation,
action on membranes,
metal ion deprivation.
Tannins induce changes in morphology of several species of ruminal bacteria.
Microrganism defense mechanisms involve
secretion of binding polymers,
synthesis of tannin-resistant enzymes,
biodegradation of tannins (peculiarity of some recently discovered bacteria that are able to tolerate high
levels of PA).

Toxicity to ruminants
Hydrolizable tannins
Hydrolizable tannins are toxic to ruminants. Tannin toxicity from HTs may occur in animals fed oak (Quercus spp.)
and several tropical tree legumes (e.g.Terminalia oblongata and Clidema hirta)
Microbial metabolism and gastric digestion convert HTs into absorbable low molecular weight metabolites. Some of
these compounds are toxic.
1.
2.

The major lesions associated with HT poisoning are hemorrhagic gastroenteritis, necrosis of the
liver, and kidney damage with proximal tuberal necrosis,
High mortality and morbidity were observed in sheep and cattle fed oaks and other tree species
with more than 20% HT.

Protanthocyanidins
Toxicity from PA is difficult to separate from their effects on the digestion of proteins and carbohydrates.
1.
2.
3.

PAs are not absorbed by the digestive tract,

PAs may damage the mucosa of the gastrointestinal tract, decreasing the absorption of nutrients,
PAs may reduce the absorption of essential aminoacids. The most susceptible amino acids are
methionine and lysine.
o Decreased methionine availability could increase the toxicity of cyanogenic glycosides,
because methionine is involved in the detoxification of cyanide via methylation to
thiocyanate.

Toxicity to monogastrics
Animals fed diets with a level of tannins under 5% experience

depressed growth rates,

low protein utilization,
damage to the mucosal lining of the digestive tract,
alteration in the excretion of certain cations, and
increased excretion of proteins and essential amino acids.
In poultry, small quantities of tannins in the diet cause adverse effects

levels from 0.5 to 2.0% can cause depression in growth and egg production,
levels from 3 to 7% can cause death.
In swine, similar harmful effects of tannins have been found.
The addition of additional proteins or amino acids may alleviate the antinutritional effects of tannins.
Levels of tannins above 5% of the diet are often lethal.
Animal defense mechanisms

Hoatzin: a ruminant-like bird that eats a lot of tannin-rich leaves

Some insects consume leaves with high levels of tannins. They are able to adapt to tannins using several available
mechanisms

alkaline gut pH,

presence of surfactants to decrease affinity between ingested tannins and protein,
presence of peritrophic membranes that absorb tannins and are then excreted in the feces.
Many tannin-consuming animals secrete a tannin-binding protein (mucin) in their saliva.

Tannin-binding capacity of salivary mucin is directly related to its proline content. Advantages in using salivary
proline-rich proteins (PRPs) to inactivate tannins are
PRPs inactivate tannins to a greater extent than do dietary proteins; this results in reduced fecal nitrogen
losses,
PRPs contain non specific nitrogen and nonessential amino acids; this makes them more convenient for an
animal to exploit rather than using up valuable dietary protein.
There are species differences in the amount of PRP that different species produce to bind tannins
Ability to tolerate tannins - deer> goat> sheep> cattle
Consumption of high tannin diets stimulates the development of the salivary glands to permit more PRP
production,
Some researchers claim that sheep and cattle do not have any PRPs.
Nutritional Effects: positive effects

The presence of tannins in food sources for monogastric animals, is generally viewed adversely, though their
contribution to red wines is certainly an exception. However, inruminants, tannins can induce beneficial effects. For
example,

In sheep and cattle higher retention of nitrogen has been observed in sheep and cattle with low to moderate levels of
tannins in forages,
In these cases, the lower apparent and true digestibility of nitrogen was compensated for by reduced urinary
loss of hydrogen,
Moderate levels of tannins (less than 4% ) in forage legumes can have beneficial responses in ruminants, resulting in
higher growth rates and milk yield,
However, even in ruminants, levels of tannins exceeding 6% of the diet result in negatively affect growth rates and
milk yield.
Several mechanisms have been suggested to explain how tannins influence protein utilization by ruminants -

Rumen escape
One mechanism postulated is that tannins complex proteins at the pH of the
rumen (5 to 7) and protect them from microbial enzymes. Subsequently,
these complexes dissociate in contact with gastric (pH 2.5-3.5) and
pancreatic (pH 8) secretions.

High quality dietary proteins would be protected, at least in part,

from degradation in the rumen and would then be digested more
effectively in the intestine. However,
o Even when released, tannins are still biologically active
and can react with digestive enzymes or other proteins.
o Indeed, in nonruminants, tannins decrease intestinal
absorption of amino acids (especially methionine) and reduce growth.
o Tannin-protein complexes that are strong enough to survive the environment of the rumen may not
be broken down and digested in the lower tract.

Urea recycling
Another hypothesis is that tannins may increase efficiency in nitrogen recycling to the rumen.
Some facts -

Tannins lower the rate of protein degradation and deamination in the rumen resulting in lower rumen
ammonia concentration.
o This results in lower plasma urea nitrogen (PUN).
Lower PUN means lower urinary nitrogen excretion with less wastage of nitrogen.
Larger amounts of nitrogen are recycled because tannins stimulate increased saliva production.

Microbial efficiency
In diets based on tanniniferous forages, nitrogen rumen outflow is often larger than nitrogen intake. Several studies
have reported an increase in protein flow when moderate doses of tannins were used. This has been attributed to -

increased rumen escape of dietary proteins,

increase in microbial protein flow (up to 28% in sheep).

The larger microbial flow could be the result of

Increased saliva production, increased rumen turnover rate, and hence, increased microbial outflow,

Increased nitrogen recycling to the rumen,

Decreased proteolysis and slower fermentation of proteins and non-protein nitrogen in the rumen
(particularly important in legume silages); this results in a more even nitrogen availability to bacteria.

Microbial flow is usually measured using a microbial internal marker (diaminopimelic acid, DAPA). However, tannins
may reduce the extraction of microbial cells walls from digesta and make microbial flows measured with DAPA
unreliable
Chemical Analysis
The amount and type of tannins synthesized by plants varies considerably depending on plant species, cultivars,
tissues, stage of development, and environmental conditions. Therefore, the study of the nutritional effects of
tannins on animals requires quantification of the tannins present in a particular diet.. Due to the complexity of
tannins, several methods have been developed for their quantification. None of them, however, is completely
satisfactory.
Sample preparation
The first factor to consider is how the forage or the feed is consumed by the animal - feeds should be analyzed in the
form eaten by the animals
If the samples are collected fresh and they have to be stored, freeze-drying is the gentlest method of preservation
and is recommended instead of freezing, air or oven-drying.

If freeze drying is too expensive or the equipment is not available, freezing without thawing of the sample
before extraction is suggested.
If drying is the only means available for preserving the material, drying temperature should be higher than
40 C (to avoid oxidation by the still active enzymes) and lower than 60 C (to avoid heat damage and
polymerization).

Sample handling
After cutting the sample should be -

Stored in a cold, dark container,

Cut in small pieces and freeze with liquid nitrogen,
Pulverized with a mortar and a pestle, and
Immediately extracted or freeze dried and stored at -4 C.

Sample extraction
Tannins are extracted with an aqueous organic solvent.

70% acetone and 30% water is a more effective extractant than alcoholic solvents.
o Acetone inhibits tannin-protein interaction. This is a limitation in protein precipitation assays.
In many plants, there is a large fraction (sometimes >50%) of the tannins that cannot be extracted (insoluble
tannins).
o This unextractable fraction cannot be ignored because of its nutritional effects.