Review: Supplementation of phytase and carbohydrases

to diets for poultry

T. A. Woyengo and C. M. Nyachoti1
Department of Animal Science, University of Manitoba, Winnipeg, Manitoba, Canada R3T 2N2.
Received 17 September 2010, accepted 30 December 2010.
Woyengo, T. A. and Nyachoti, C. M. 2011. Review: Supplementation of phytase and carbohydrases to diets for poultry. Can.
J. Anim. Sci. 91: 177
192. Feedstuffs of plant origin contain anti-nutritional factors such as phytic acid (PA) and non-
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 105.104.230.235 on 05/17/20

starch polysaccharides (NSP), which limit nutrient utilization in poultry. Phytic acid contains phosphorus, which is poorly
digested by poultry, and has the capacity to bind to and reduce the utilisation of other nutrients, whereas NSP are
indigestible and have the capacity to reduce nutrient utilisation by encapsulation. Supplemental phytase and NSP-
degrading enzymes (carbohydrases) can, respectively, hydrolyze PA and NSP, alleviating the negative effects of these anti-
nutritional factors. In feedstuffs of plant origin, PA is located within the cells, whereas NSP are located in cell walls, and
hence it has been hypothesized that phytase and carbohydrases can act synergistically in improving nutrient utilization
because the carbohydrases can hydrolyze the NSP in cell walls to increase the accessibility of phytase to PA. However, the
response to supplementation of a combination of these enzymes is variable and dependent on several factors, including the
type of carbohydrase supplement used, dietary NSP composition, calcium and non-phytate phosphorus contents, and
endogenous phytase activity. These factors are discussed, and areas that need further research for optimising the use of a
combination of phytase and carbohydrases in poultry diets are suggested.

Key words: Phytase, carbohydrases, nutrient utilization, poultry

Woyengo, T. A. et Nyachoti, C. M. 2011. Rations enrichies de phytase et de carbohydrases pour la volaille. Can. J. Anim.
Sci. 91: 177
192. Les aliments d’origine végétale renferment des facteurs antinutritionnels comme l’acide phytique (PA) et
For personal use only.

des polysaccharides non amylacés (PNA), qui restreignent l’assimilation des éléments nutritifs chez la volaille. L’acide
phytique contient du phosphore que digère mal la volaille et qui peut se lier à d’autres éléments nutritifs pour en réduire
l’assimilation, tandis que les PNA ne peuvent être digérés et diminuent l’assimilation des autres nutriments par
encapsulation. Les suppléments de phytase et d’enzymes dégradant les PNA (carbohydrases) peuvent, respectivement,
hydrolyser l’acide phytique et les PNA, et en atténuer les incidences négatives sur la nutrition. Dans les aliments d’origine
végétale, l’acide phytique se trouve à l’intérieur des cellules, alors que la PNA se situe dans la paroi cellulosique. On a donc
formulé l’hypothèse que la phytase et les carbohydrases pourraient agir en synergie et rehausser l’assimilation des éléments
nutritifs, les seconds hydrolysant les PNA de la paroi cellulosique pour permettre à la phytase d’accéder plus aisément à
l’acide phytique. Néanmoins, la réaction à un supplément combinant ces deux enzymes varie et dépend de plusieurs
facteurs parmi lesquels figurent le type de carbohydrase employé, la composition des PNA dans l’aliment, la teneur en
calcium et en phosphore non-phytate, et l’activité endogène de la phytase. Les auteurs discutent de ces facteurs et suggèrent
les aspects sur lesquels il faudrait entreprendre des recherches plus poussées en vue d’optimiser la combinaison de la
phytase et des carbohydrases dans la ration de la volaille.

Mots clés: Phytase, carbohydrases, assimilation des éléments nutritifs, volaille

Phosphorus is the third most expensive component of
poultry diets after energy and amino acids. Thus, there is
need to optimize the utilization of P by poultry to reduce
the cost of feeding. Feedstuffs of plant origin constitute a
high proportion of poultry feeds because they are cheap-
er than their animal counterparts. However, approxi-
mately two-thirds of P in vegetable feedstuffs is poorly
digested by poultry because it is bound to phytic acid
(PA), which is poorly hydrolyzed by poultry, as they do
not produce sufficient amounts of phytase enzyme that
hydrolyses PA (Bedford 2000). Because of the low
capacity of poultry to digest PA-bound P, inorganic
1
Corresponding author (martin_nyachoti@umanitoba.ca).
sources of P, which are expensive, are added to feeds to
meet the P requirements of poultry, leading to increased
cost of feeding (Selle and Ravindran 2007). Furthermore,
PA can reduce the utilisation of nutrients other than P by
binding to them (Lenis and Jongbloed 1999). Thus, the
presence of PA in poultry diets can result in reduced
efficiency of nutrient utilization and hence increased cost
of feeding, and environmental pollution.
In addition to PA, feedstuffs of plant origin also
contain non-starch polysaccharides (NSP) in their cell
wall, which are indigestible and capable of reducing
Abbreviations: AA, amino acids; AME, apparent metabolisable
energy; FTU, fytase unit; NSP, non-starch polysaccharides; PA,
phytic acid

Can. J. Anim. Sci. (2011) 91: 177
192 doi:10.4141/CJAS10081 177

 178 CANADIAN JOURNAL OF ANIMAL SCIENCE

utilisation of nutrients in poultry by limiting the avail-

ability for digestion and absorption (Bedford and
Schuzle 1998; Kim et al. 2005). The presence of NSP
in poultry diets can thus also result in reduced efficiency
of nutrient utilisation.
Supplementation of poultry diets with phytase and
NSP-degrading enzymes (carbohydrases) may alleviate
the anti-nutritional effects that are associated with PA
and NSP, respectively. The NSP can limit the accessi-
bility of phytase to PA (Kim et al. 2005). Thus, phytase
and carbohydrases may act synergistically in improving
nutrient utilisation in poultry because carbohydrases can
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hydrolyze NSP to increase the accessibility of phytase to

PA. The effect of supplementing phytase and carbohy-
drases individually to poultry diets on nutrient utilisation
has been evaluated in several studies and it has been
reviewed (Bedford 1995, 2000; Bedford and Schuzle
1998; Adeola and Sands 2003; Selle et al. 2006;
Selle and Ravindran 2007). However, information on
the effect of combining phytase and carbohydrases on Fig. 1. Phytic acid.
performance and nutrient digestibility in poultry has not
been reviewed. The objective of this paper is to review embryonic cells. Its concentration in various feed
results from studies on the effects of combining phytase ingredients that are commonly used in poultry feeds
and carbohydrases on nutrient utilisation in poultry, and also varies depending on several factors, including type
to discuss factors that influence the combination effects and variety of ingredient, and year of harvesting (Steiner
of phytase and carbohydrases on nutrient utilization in et al. 2007). It is, however, generally higher in cereal
For personal use only.

poultry. Also, areas that need further research for milling by-products and oil seed meals than in grain
optimising the use of a combination of phytase and legumes and cereal grains as indicated by phytate-P
carbohydrases in poultry diets are suggested. concentration in these feed ingredients (Table 1). Its
high concentration in cereal by-products is due to its
I. PHYTIC ACID high concentration in the aleurone cells of cereal grains,
Phytic acid is a constituent of plant seeds in which it which form part of by-products from these grains
serves as the major storage form of P (Maenz 2001). (Steiner et al. 2007).
Chemically, it consists of an inositol ring with 6 phos-
phate groups and 12 protons, i.e., 2 protons per Effect of Phytic Acid on Mineral Digestibility
phosphate group (Fig. 1; Maenz 2001; Cowieson et al. Poultry digest the PA-bound P poorly because they do
2004). Of the 12 protons, 6 dissociate at a pKa value of not produce sufficient amount of phytase (Kornegay
approximately 1.5, 3 at pKa values between 5.7 and 7.6, 2001). For example, Rutherfurd et al. (2004) and
and the remaining three at pKa values greater than 10, to Cowieson et al. (2006a) found the digestibility of PA-
leave a negatively charged PA at these acidic, neutral and bound P in broilers to be as low as 10%. The unabsorbed
basic pH conditions (Maenz 2001). Phytic acid occurs as P is excreted via feces, which can lead to environmental
phytate (i.e., a mixed salt of cations, mainly K and Mg,
and to a lesser extend Ca, Fe and Zn) in spherical Table 1. Mean concentration of phytate-P and phytase activity of
inclusions called globoids, which are located within common feed ingredientsz
protein bodies (Ockenden et al. 2004; Joyce et al. 2005;
Ingredient Phytate P (g kg 1) Phytase activity (FTU kg 1)
Lin et al. 2005) in cereal grains, grain legumes and
oilseeds that constitute the majority of ingredients in Wheat 2.2, 2.9 1193, 2886
poultry feeds. In soyabean, however, most of the phytate Barley 2.2, 2.6 582, 2323
(85 to 90%) in protein bodies does not occur as spherical Triticale 2.5, 2.8 1688, 2799
Rye 2.2, 2.4 5130, 6016
inclusions; it interacts directly with the protein (Prattley Corn 2.1 B50
and Stanely 1982). Sorghum 2.4 B50
Phytic acid is not uniformly distributed in plant seeds, Wheat bran 7.9 2957, 9945
but localized within certain seed components. For Rye bran 4.9 9241
instance, in wheat (Joyce et al. 2005) and barley Soyabean meal 4.5 B50
Canola meal 6.7 B50
(Ockenden et al. 2004) phytate is concentrated in the
aleurone cells, whereas in corn (Lin et al. 2005) and z
Two values separated by a comma were obtained from two different
oilseeds like soyabean (Prattley and Stanely 1982) and sources; sources of data
Eeckhout and De Paepe (1994), Selle et al.
canola (Yiu et al. 1982) it is concentrated in the (2003a) and Steiner et al. (2007).
 WOYENGO AND NYACHOTI * PHYTASE AND CARBOHYDRASES FOR POULTRY DIETS
pollution (Knowlton et al. 2004; Toth et al. 2006). Phytic
acid can also reduce the digestibility of other minerals
because in its natural state in plants, it is complexed with
minerals in globoids within protein bodies (Ockenden et
al. 2004; Joyce et al. 2005; Lin et al. 2005). Furthermore,
because of its negative charges at acidic, neutral and
basic pH conditions, PA can complex cations at all pH
conditions in the gastro-intestinal tract, thereby reducing
the bioavailability of the cations, especially divalent
cations, which form insoluble complexes with PA at
small intestinal pH conditions (Maenz et al. 1999).
Several studies on the effect of PA on mineral
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digestibility have indeed shown a negative relationship
between dietary PA concentration and mineral digest-
ibility. Ravindran et al. (2006) reported that an increase
in the level of PA concentration in diets from 10.4 to 13.6
g kg 1 resulted in reduced ileal digestibility of Ca and Fe
from 37.7 to 36.0% and from 21.8 to 20.3%, respectively.
Urbano et al. (1999) similarly reported decreased Ca
digestibility from 82.1 to 59.8% in rats fed lentil-based
diets due to an increase in dietary PA concentration from
4.77 to 6.46 mg g1. In pigs, we have observed reduced
apparent ileal digestibility of Ca, Mg, Na and K in
piglets by a mean of 38 percentage points due to sup-
plementation of a casein-cornstarch-based diet with PA
at 2% (Woyengo et al. 2009). Kemme et al. (1999) also
For personal use only.
reported reduced ash digestibility in growing pigs from
56.9 to 54.7% due to an increase in dietary PA
concentration from 9.4 to 14.3 g kg1.
Effect of Phytic Acid on Digestibility of Amino
Acids and Energy
The digestibility of amino acid (AA) and energy, like that
of minerals, is negatively affected by PA. Ravindran
et al. (2000), feeding broilers on wheat-sorghum-
soyabean meal-based diets, reported a decrease in ileal
digestibility of nitrogen of 2.3 percentage points due to
an increase in dietary PA concentration from 10.4 to 15.7
g kg1. In their study, the ileal digestibility of all the
essential AA was also reduced by the increase in dietary
PA concentration. More recently, Cowieson et al.
(2006a) precision fed growing broilers on 5 g of casein
for 48 h and observed a decrease in total AA digestibility
by 8.9 and 11.9% due to addition of 0.5 and 1 g of PA to
5 g of casein, respectively. An increase in PA concentra-
tion in corn-soyabean meal-based diets for broilers from
10.0 to 13.6 g kg 1 by the addition of rice bran has also
been reported to result in a decrease in apparent
metabolizable energy (AME) value of the diet by 2.1%
(3353 vs. 3281 kcal kg 1; Ravindran et al. 2006).
A mechanism by which PA can adversely affect AA
and energy digestibility before feed ingestion has been
proposed. Phytic acid in its natural state is complexed
with AA in protein bodies (Ockenden et al. 2004; Joyce
et al. 2005; Lin et al. 2005) and thus it can minimize the
availability of these nutrients for digestion and absorp-
tion (Lenis and Jongbloed 1999). Also, three mechan-
isms by which PA can adversely affect AA and energy
179
digestibility after feed ingestion have been proposed.
First, the terminal amino groups on proteins and free
amino groups on basic AA possess positive charges at
pH below the iso-electric point, i.e., at pH less than 5
(Prattley et al. 1982; Maenz 2001). Consequently, at
acidic pH in the pro-ventriculus and ventriculus of
poultry, PA can interact with protein directly to form
electrostatic bonds (Prattley and Stanely 1982), thereby
reducing the availability of protein for digestion in the
pro-ventriculus and ventriculus of poultry (Cowieson
et al. 2004). This hypothesis has been confirmed by an in
vitro study by Kies et al. (2006), who incubated 10 g of
casein with or without 1 mg of PA and observed its
(casein) complete dissolution at pH 2 in the absence of
PA, and almost complete precipitation (99%) in the
presence of PA.
Second, protein possesses a net negative charge at a
pH above the isoelectric point, but below pH 10 (Maenz
2001). Hence, in the small intestine, where the pH is
within the range of 5 to 10, PA can bind protein through
multivalent cations to form a PA-mineral-protein com-
plex that is resistant to enzymatic hydrolysis (Maenz
2001; Adeola and Sands 2003). This ability of PA to bind
protein via cations was clearly demonstrated by Prattley
et al. (1982), who incubated serum albumin with PA
either in the presence or absence of Ca, and observed the
formation of PA
protein complexes only in the presence
of Ca. Third, endogenous proteolytic enzymes, like any
other protein, can be bound by PA in both the stomach
and small intestine, thereby reducing their ability to
digest dietary proteins (Cowieson et al. 2006a; Selle et al.
2006). For instance, Singh and Krikorian (1982) mea-
sured the activity of trypsin in vitro at pH 7.5 using
casein as a substrate and reported an increase in
inhibition of enzyme activity from 2.7 to 42.5% (after
incubation for 20 min) due to an increase in the level of
phytate concentration from 10 to 90 mM. We also
observed a 46% reduction in pepsin activity in piglets
from supplementation of a casein-cornstarch-based diet
with PA at 2% (Woyengo et al. 2010a).
Direct binding of energy-generating molecules such
as carbohydrates, lipids and protein by PA can reduce
energy digestibility (Thompson et al. 1987; Selle and
Ravindran 2007). Phytic acid can also reduce energy
digestibility indirectly by binding endogenous enzymes
and metal cofactors of enzymes involved in hydrolysis of
energy generating molecules (Thompson et al. 1987),
and by increasing the endogenous secretion of Na,
which is involved in AA and glucose absorption from
the gastro-intestinal tract (Woyengo et al. 2009).
II. PHYTASES
Phytases are enzymes that catalyze step-wise cleavage of
phosphate groups from PA (Selle and Ravindran 2007).
The site of initial removal for some phytases is the
phosphate group at the C6 position on the inositol ring,
whereas for others it is the phosphate group at position
C3 (Fig. 1) (Kornegay 2001). Phytases, whose site of
 180 CANADIAN JOURNAL OF ANIMAL SCIENCE

initial hydrolysis is the phosphate group at the C6
position are known as 6-phytases, whereas those whose
initial site of initial hydrolysis is the phosphate group at
the C3 position are known as 3-phytases (Selle and
Ravindran 2007). Their preferred molecule for the
removal of the phosphate group is, however, a fully
phosphorylated PA followed by penta-, tetra-, tri-, di
and mono-esters of inositol in descending order of
preference (Wyss et al. 1999; Lassen et al. 2001; Vats
and Banerjee 2004). They thus normally hydrolyze all
the available fully phosphorylated PA molecules to
penta-esters of inositol before they hydrolyze the latter
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(Maenz 2001; Pandey et al. 2001). Among micro-
organisms, the major phytase producers are fungi, yeast
and bacteria (Table 2; Pandey et al. 2001). Most
phytases produced by these micro-organisms are 3-
phytases (Maenz 2001) except for a few like Basidiomy-
cete fungi- (Lassen et al. 2001) and Escherichia coli
bacteria- (Greiner et al. 1993) derived phytases, which
are 6-phytases.
In plants, phytases occur in seeds, where their major
role appears to be the release of P from PA during
germination for utilisation by the developing plant
(Centeno et al. 2001). The plant phytases are 6-phytases

to tetra-esters of inositol and so on (Wyss et al. 1999).
Phytases are normally inhibited by their product
(inorganic P; Greiner et al. 1993; Hu et al. 1996; Greiner
2002; Lopez et al. 2000), and like other enzymes, they
have pH and temperature ranges at which they function
optimally (Greiner and Konietzny 2006). Their activity
is measured in fytase units; where 1 fytase unit is the
amount of phytase that liberates 1 mmol of inorganic
phosphate per minute from 0.0051 mol L 1 sodium
phytate at pH 5.5 and at temperature of 378C (Engelen
et al. 1994). Fytase units are commonly abbreviated
as FTU, though other abbreviations including FYT,
U, and PU have also been used (Selle and Ravindran
2007). In the current paper, FTU is used to denote fytase
For personal use only.
(Maenz 2001; Schlemmer et al. 2001), and among
feedstuffs that are commonly used in formulation of
poultry feeds, the activity of phytase is higher in
‘‘viscous grains’’ such as wheat, rye and barley, and
their milling by-products than in corn, sorghum and
oilseed meals (Table 1).
Plant phytases have been shown to significantly
hydrolyze PA in poultry (Paik 2003) and pigs (Rapp
et al. 2001). They are, however, not as effective as
microbial phytase. For instance, PA hydrolysis in the
stomach of minipigs fed a diet supplemented with the
microbial (Aspergillus niger) phytase at 818 FTU kg1
was found to be 17% higher than in those fed a diet with
supplemented plant (wheat) phytase at 1192 FTU kg1

units.
Sources of Phytases
Phytases are produced to a greater extent by micro-
organisms and plants, and to a lesser extent by animals
(Rapp et al. 2001). The recovery of the wheat phytase in
the duodenum of the same animals was also lower than
that of Aspergillus niger phytase (45 vs. 70%; Rapp
et al. 2001). Phillippy (1999) has similarly reported a
higher recovery of A. niger phytase than wheat phytase

Table 2. Sources and properties of phytases

Optimum
Origin Expression host temperature (8C) Optimum pH Reference

Fungi
Aspergillus. niger

2.5
4.5 Tomschy et al. (2002)
A. niger PhyA A. oryzae 50 2.5
5.5 Lassen et al. (2001)
A. fumigates A. fumigates
4.0, 6.0
6.5 Pasamontes et al. (1997)
A. ficuum A. niger 58 5.5, 2.5 Ullah et al. (2000)
A. pediades PhyA A. oryzae 50 5.0
6.0 Lassen et al. (2001)
Peniophora lycii

4.0
4.5 Tomschy et al. (2002)
P. lycii A. oryzae 58 3.0 Ullah et al. (2000)
P. lycii PhyA A. oryzae 50
55 4.0
4.5 Lassen et al. (2001)
Ceriporia sp. PhyA1 A. oryzae 55
60 5.5
6.0 Lassen et al. (2001)
Ceriporia sp. PhyA2 A. oryzae 40
45 5.5
6.0 Lassen et al. (2001)
Rhizopus oligosporus
65 5.0 Casey and Walsh (2004)
Bacteria
Escherichia coli
55 4.5 Greiner et al. (1993)
E. coli Bacillus subtilis
2.0
4.5 Adeola et al. (2004)
E. coli Schizosaccharomyces pombe 55 3.5 Seonho et al. (2005)
Klebsiella sp. strain ASR1 E. coli 45 5.0 Sajidan et al. (2004)
Obesumbacterium proteus E. coli 40
50 4.9 Zinin et al. (2004)
Plants
Barley (PI)
55 6.0 Sung et al. (2005)
Barley (PII)
50 5.0 Sung et al. (2005)
Wheat
50 5.0
6.0 Paik (2003)
Lupinus albus var. Amiga
50 5.0 Greiner (2002)
 WOYENGO AND NYACHOTI * PHYTASE AND CARBOHYDRASES FOR POULTRY DIETS
(95 vs. 70%) after 1 h of preincubation with 5 mg of
pepsin mL 1.
The poorer performance of plant phytases compared
with microbial phytases is due to their narrower
optimum pH range (Greiner and Konietzny 2006). The
optimum pH for plant phytases ranges from 5.0 to 6.0,
whereas that for A. niger, Peniophora lycii and Escher-
ichia coli derived phytases, the most commonly utilized
microbial phytses (Lei and Stahl 2001; Greiner and
Konietzny 2006; Selle and Ravindran 2007), ranges
from 2.0 to 5.5 (Table 2), which is closer to the physio-
logical pH (2 to 5) of the pro-ventriculus and ventriculus
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of poultry (Simon and Igbasan 2002). The poorer
performance of plant phytases compared with microbial
phytases is also attributed to greater resistance of the
latter to protease enzymes in the gastro-intestinal tract
due to a higher degree of glycosylation (Phillippy 1999).
Animal phytases are produced by intestinal mucosa
(Yang 1991; Applegate et al. 2003). Their activity has
been found to be higher in the duodenum than in other
intestinal segments of rats (Lopez et al. 2000) and
poultry (Maenz and Classen 1998) and their production
appears to be induced by the presence of its substrate
(PA) in the intestine (Lopez et al. 2000) and reduced by
increased concentration of its product (inorganic P) in
the same gastro-intestinal tract compartment (Hu et al.
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1996).
Intestinal phytase has been reported to increase PA
hydrolysis by 24% in rats fed a purified diet after
intestinal perfusion with pure PA for 20 d (Lopez et al.
2000). Its effect on the hydrolysis of PA in conventional
(unpurified) diets has, however, been reported to be
negligible (Jongbloed et al. 1992; Rapp et al. 2001), and
it is attributed to the failure of PA to induce mucosal
phytase production since, in conventional feeds, it
occurs as phytate in protein bodies and thus it rarely
comes in contact with the intestinal mucosa (Lopez et al.
2000).
Phytases for the Poultry Feed Industry
Because of their ability to liberate P from PA, phytases
are commercially produced and added to poultry feeds to
improve nutrient digestibility and utilization. An ideal
phytase for the poultry feed industry would be the one
that is resistant to acidic pH and protease enzymes in the
stomach and small intestine, where P absorption takes
place, is cost-effective to produce, and is resistant to high
temperatures (65 to 808C) that are encountered during
feed pelleting (Lei and Stahl 2001). Because microbial
phytases compared with other phytases have a greater
ability to hydrolyze PA in the gastro-intestinal tract, a lot
of research has been focused on the identification and
testing of efficacy of the former for use in the animal feed
industry (Pandey et al. 2001). It has, however, been
difficult to obtain native microbial phytases with all the
above-mentioned attributes (Lei and Stahl 2001). Thus,
several microbial phytases have been modified by proces-
ses such as genetic transformation and thermo-protective
181
coating in an effort to achieve these desired character-
istics (Lei and Stahl 2001; Garrett et al. 2004; Barletta
2007). Currently, most of the phytases that are commer-
cially available to the feed industry are derived from
micro-organisms, especially fungi and bacteria (Lei and
Stahl 2001; Selle and Ravindran 2007). Thus, this review
will focus on the microbial phytases.
Sites of Supplemental Microbial Phytase
Activity in the Digestive Tract
Supplemental microbial phytases hydrolyze PA in some,
but not all, gastro-intestinal tract compartments in
poultry. Yu et al. (2004) observed the highest activity of
a fungal (P. lycii) phytase in the crop and ventriculus,
followed by the duodenum and jejunum, and negligible
activity in the ileum. Onyango et al. (2005), feeding
broilers on diets supplemented with either bacterial
(E. coli) or P. lycii phytase at 1000 FTU kg 1, observed
the highest activity of P. lycii phytase in the crop (404
FTU kg1) followed by the gizzard (63 FTU kg 1),
and negligible activity in the jejunum (25 FTU kg1)
and ileum (6 FTU kg1). For E. coli phytase, however,
although the activity was highest in the crop (649 FTU
kg 1), it remained relatively high in the pro-ventriculus
and ventriculus combined (406 FTU kg 1) and jejunum
(554 FTU kg1), and was only low in the ileum (91 FTU
kg 1). The major sites of activity of supplemental fungal
phytases are the crop and the ventriculus in poultry,
because they have maximal activity at the acidic pH that
is within the pH range present in these gastro-intestinal
tract regions and are susceptible to proteolysis that
occurs in the small intestine (Simon and Igbasan 2002).
When compared with fungal phytase, E. coli phytase
remains active up to the jejunal region because it is
more resistant to the proteolysis that occurs in the small
intestine (Onyango et al. 2005).
Effect of Supplemental Microbial Phytase on
P Utilization
Phytase supplementation has been shown to improve P
digestibility in poultry (Table 3). Because of this im-
proved digestibility of P due to phytase, the non-phytate
P in poultry diets has been reduced by 0.10 to 0.20
percentage points without any significant effect on
performance (Table 4). Phytase supplementation has
also been shown to increase the digestibility of other
minerals and AA, and AME values (Ravindran et al.
2001; Onyango et al. 2004; Silversides et al. 2004;
Cowieson et al. 2006b; Ravindran et al. 2006; Selle
et al. 2009). However, based on the observed improve-
ment in ileal P digestibility of 7.2 to 20.6 percentage
points by phytase in the studies presented in Table 3,
phytase hydrolyzed 20 to 65% of phytate P in diet,
which is incomplete and variable hydrolysis. Some of the
factors that could affect the efficacy of phytase with
regard to PA-P hydrolysis include dietary level of
inorganic (non-phytate) P and Ca, dietary endogenous
phytase activity and dietary non-starch polysaccharides
 182 CANADIAN JOURNAL OF ANIMAL SCIENCE

Table 3. Results from selected studies on the effect of phytase on ileal phosphorus digestibility in broilers

Diet type Phytase level (FTU kg 1) Increase in P digestibility (percentage points) Reference

Corn-soyabean meal 500 15.7 Dilger et al. (2004)

Corn-soyabean meal 750 11.8 Rutherfurd et al. (2004)
Corn-soyabean meal 1000 16.8 Ravindran et al. (2006)
Corn-soyabean meal 600 13.5 Woyengo et al. (2010b)
Corn-soyabean meal 700 20.5 Emiola et al. (2007)
Corn-soyabean meal 1000 20.6 Olukosi et al. (2007)
Corn-soyabean meal 1000 20.6 Tiwari et al. (2010)
Wheat-soy protein 600 11.8 Afsharmanesh et al. (2008)
Wheat-soyabean meal 500 7.2 Woyengo et al. (2008)
Wheat-soyabean meal 500 14 Selle et al. (2009)
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 105.104.230.235 on 05/17/20

composition. The effects of these factors on the efficacy III. NON-STARCH POLYSACCHARIDES
of phytase are subsequently discussed in Section VI Non-starch polysaccharides are complex carbohydrates
below. other than starch that are naturally found in plant cell

Table 4. Results from selected studies on the effect of supplementing low-P diets with phytase on performance of poultry

Poultry Non-phytate P in Performance for Performance for BD

type Basal dietz BDy (%) Response criterion BD vs. PCx phytasew vs. PC Reference

Broiler Corn-SBM 0.45 BWG Similar Similar Powell et al. (2008)

Broiler Wheat-SBM 0.45 BWG Similar Similar Wu et al. (2003)
Broiler Corn-SBM 0.40 BWG Similar Similar Powell et al. (2008)
Broiler Corn-SBM 0.35 BWG Lower Similar Angel et al. (2006)
For personal use only.

Broiler Corn-SBM 0.34 BWG Lower Similar Olukosi et al. (2007)

Broiler Corn-SBM 0.31 BWG Lower Similar Pillai et al. (2006)
Broiler Wheat-SBM 0.30 BWG Lower Similar Wu et al. (2003)
Broiler Wheat-SBM 0.26 BWG Lower Similar Selle et al. (2009)
Broiler Corn-SBM 0.24 BWG Lower Similar Dilger et al. (2004)
Broiler Corn-SBM 0.24 BWG Lower Similar Onyango et al. (2004)
Broiler Wheat-SBM 0.23 BWG Lower Similar Silversides et al. (2004)
Broiler Corn-SBM 0.22 BWG Lower Lower Angel et al. (2006)
Broiler Wheat-SBM 0.15 BWG Lower Lower Zyla et al. (1999a)
Broiler Corn-SBM 0.12 BWG Lower Lower Dilger et al. (2004)
Laying Corn-SBM 0.25
0.20
0.15v BWG, egg production Similar Similar Keshavarz (2000a)
hens
Laying Corn-SBM 0.20
0.15
0.10u BWG, egg production Lower Similar Keshavarz (2000a)
hens
Laying Corn-SBM 0.15
0.10
0.10t BWG, egg production Lower Similar Keshavarz (2000a)
hens
s
Laying Wheat-SBM 0.20
0.15 BWG, egg production Similar Similar Silversides et al. (2006)
hens
Growing Corn-SBM 0.30
0.25
0.20r BWG Similar Similar Keshavarz (2000b)
pullets
q
Laying Corn-SBM 0.10 Egg production Lower Similar Gordon and Roland (1997)
hens
Laying Corn-SBM 0.15p Egg production Lower Similar Liu et al. (2007)
hens

z
SBMsoyabean meal.
y
BDbasal diet without phytase.
x
PC positive control (non-phytate P adequate) diet; Similargrowth performance of broilers fed the basal diet without phytase was similar to that
of broilers fed the positive control diet.
w
BDphytasebasal diet plus phytase; Similargrowth performance of broilers fed the basal diet plus phytase was similar to that of broilers fed
the positive control diet; Lowergrowth performance of broilers fed the basal diet plus phytase was lower than that of broilers fed the positive
control diet.
v
Each non-phytate P sequence was used for 30 to 42, 42 to 54, and 54 to 66 wk of age, respectively.
u
Each non-phytate P sequence was used for 30 to 42, 42 to 54, and 54 to 66 wk of age, respectively.
t
Each non-phytate P sequence was used for 30 to 42, 42 to 54, and 54 to 66 wk of age, respectively.
s
Each non-phytate P sequence was used for 34 to 49 and 50 to 64 wk of age, respectively.
r
Each non-phytate P sequence was used for 0 to 6, 6 to 12, and 12 to 18 wk of age, respectively.
q
Hens were fed the diets from week 21 to wk 38 of age.
p
Hens were fed the diets from week 23 to wk 28 of age.
 WOYENGO AND NYACHOTI * PHYTASE AND CARBOHYDRASES FOR POULTRY DIETS
walls (McDonald et al. 1995). Some of the NSP found
in feedstuffs that are commonly used to formulate
poultry diets include cellulose, b-glucans, arabinoxylans,
pectins, gums and mucillages (McDonald et al. 1995).
Non-starch polysaccharides are indigestible by poultry
(Bedford 2000), and they can reduce the availability of
nutrients within the cells by encapsulation (Bedford
and Schuzle 1998). Furthermore, soluble b-glucans and
arabinoxylans can increase digesta viscosity, which,
in turn, can reduce nutrient intake and digestibility
(Bedford and Schulze 1998).
Several studies have been conducted to evaluate the
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 105.104.230.235 on 05/17/20
anti-nutritive effects of NSP. Zyla et al. (1999b) reported
a positive correlation between arabinoxylans concentra-
tion and viscosity of wheat digested in vitro, whereas
Carre et al. (2002) found negative correlation between in
vitro viscosity and AME of wheat in broilers. Boros
et al. (2002) observed a 9% reduction in FCR and fat
digestion of broilers due to an increase in dietary
concentration of soluble arabinoxylans from 3.36 to
7.74%. In pigs, Zijlstra et al. (1999) found a negative
correlation between digestible energy in growing pigs
and total NSP, insoluble NSP and xylose concentrations
in wheat-based diets. Soluble NSP concentration in the
diets was, however, not related to the digestible energy
indicating that viscosity might not be a problem in pigs.
For personal use only.
IV. NON-STARCH POLYSACCHARIDES
DEGRADING ENZYMES
Carbohydrases are enzymes that hydrolyze NSP into
oligo-saccharides and monosaccharides (Bedford 2000).
These carbohydrases are not produced by vertebrates
such as poultry, but by plants and micro-organisms with
the purpose of providing sugars (Bedford and Schulze
1998). Because of their ability to hydrolyze NSP, NSP
degrading carbohydrases are commercially produced
and added to poultry feeds to improve nutrient digest-
ibility and performance.
The types and concentrations of NSP vary among
feedstuffs. Thus, the types of carbohydrases that are
supplemented to diets to improve nutrient availability
have been determined based on the dietary NSP compo-
sition. For instance, wheat, rye, triticale and corn are rich
in arabinoxylans, whereas barley and oats are rich in b-
glucans (Bedford 1995; Choct 1997). Hence, xylanases
are supplemented to wheat-, rye-, triticale- and corn-
based diets, whereas b-glucanases are supplemented to
barley- and oat-based diets with the view of improving
nutrient digestibility and performance.
Several studies have shown improved nutrient utiliza-
tion in poultry due to carbohydrase supplementation.
For instance, Wu et al. (2004b) observed increased
AME, body weight gain and feed conversion ratio of
3-wk-old broilers fed wheat-based diets by 1.1, 2.6 and
1.5%, respectively, due to supplementation of the diets
with xylanase at 1000 XU kg 1. The duodenal, jejunal
and ileal digesta viscosities were also reduced by 11.3,
20.9 and 18.1%, respectively. Lazaro et al. (2003)
183
supplemented rye-based diets for broilers (4 to 25 d)
with an enzyme preparation containing 858 units of
b-glucanase and 864 units of xylanase g1 at a rate of
500 ppm to determine the effect of enzyme on digesta
flow rate in the gastro-intestinal tract and performance
of broilers. Enzyme supplementation reduced the time
required to recover 1% (0.78 vs. 0.98 h) and 50% (4.2 vs.
6.5 h) of the marker in feces. Enzyme supplementation
also improved weight gain (31.7 vs. 38.3 g d1), feed
intake (63.6 vs. 66.7 g d1) and FCR (1.96 vs. 1.71
g g1) and reduced intestinal viscosity from 321 to 86
cps. Xylanase supplementation has also been found to
improve the pancreatic lipase and chymotrypsin activ-
ities in broilers fed wheat-based diets by 23 and 9%,
respectively (Engberg et al. 2004).
Some studies have, however, not shown a beneficial
effect of carbohydrases. For example, Preston et al.
(2000) observed a non-significant effect of xylanase on
the performance of broilers fed wheat-based diets
supplemented with xylanase at 2500 XU kg 1. We
also did not observe an improvement in growth perfor-
mance or nutrient digestibility of broilers fed wheat-
based diets due to the addition of xylanase to the diets at
1250 or 2500 XU kg1 (Woyengo et al. 2008). This
variable response to carbohydrase supplementation is
probably due to several factors, including type of sup-
plemental carbohydrase and dietary NSP concentration,
which are discussed later in Section VI.
V. EFFECT OF COMBINING PHYTASE AND
CARBOHYDRASES
In feedstuffs of plant origin, PA is located within the
cells (Prattley and Stanley 1982). However, NSP in cell
walls of the feedstuffs can reduce the availability of cell
contents by encapsulation (Bedford and Schuzle 1998).
Hence, the presence of NSP in vegetable feedstuffs can
limit the accessibility of phytase to PA, leading to
reduced PA degradation (Kim et al. 2005). Furthermore,
the soluble NSP can increase digesta viscosity, which, in
turn, can decrease the absorption of nutrients released
by the action of phytase. Supplementation with carbo-
hydrases, can improve the availability of nutrients
within the cells and reduce digesta viscosity (Bedford
2000), implying that carbohydrases can also increase the
efficacy of phytase by increasing the accessibility of
phytase to PA and absorption of nutrients released by
phytase. Several studies have shown that supplementa-
tion of a combination of phytase and carbohydrases to
corn-, wheat- or barley-based diets for poultry is more
beneficial with regard to nutrient utilization than
supplementation of the enzymes individually. We have
observed greater body weight gain by broilers fed low-P
corn-based diet from day 1 to day 22 of age after
supplementation of a combination of phytase and a
cocktail of carbohydrases (xylanase, b-glucanase, pecti-
nase, cellulase, mannanase and galactanase) than
after supplementation of phytase alone (673 vs. 632 g;
Woyengo et al. 2010b).
 184 CANADIAN JOURNAL OF ANIMAL SCIENCE
For wheat-based diets, Selle et al. (2009) observed
increased ileal digestibility of AA in broilers fed low-P
wheat-based diet by an average of 8.6% when phytase
and xylanase were supplemented in combination, which
was higher than the increases of 4.8 and 5.5%
for xylanase and phytase, respectively, when the
two enzymes were supplemented individually. Also,
Ravindran et al. (1999) reported a greater improvement
in overall ileal digestibility of AA in broilers when
phytase and xylanase were supplemented in combina-
tion than when the two enzymes were supplemented
alone, whereas Zyla et al. (1999a) reported greater
Can. J. Anim. Sci. Downloaded from www.nrcresearchpress.com by 105.104.230.235 on 05/17/20
improvement in body weight gain in broilers when
phytase and xylanase were supplemented in combina-
tion than when the two enzymes were supplemented
individually. For barley-based diets, Juanpere et al.
(2005) observed that supplementation of a combination
of phytase and b-glucanase improved P retention in
broilers by 15%, which was greater than improvement
observed for phytase (1.6%) alone; b-glucanase alone
did not improve P retention.
Several studies have, however, not shown any bene-
ficial effect of combining phytase and carbohydrases on
nutrient utilization in poultry. Selle et al. (2003b), Wu
et al. (2004a), Juanpere et al. (2005), Olukosi and Adeola
(2008), and Woyengo et al. (2008) did not observe any
For personal use only.
synergistic interaction between phytase and xylanase on
P, Ca or AA digestibilities, or performance of broilers
fed wheat-based diets. Silversides et al. (2006) similarly
observed no synergistic interactions between phytase and
xylanase on egg production by laying hens fed wheat-
based diets. Also, Cowieson and Adeola (2005), Juan-
pere et al. (2005), Olukosi et al. (2007, 2008), Lu et al.
(2009) and Tiwari et al. (2010) did not observe synergistic
interactions between phytase and carbohydrases on P,
Ca or AA digestibilities, or performance of broilers fed
corn-based diets. In all these studies in which there were
no synergistic interactions between phytase and carbo-
hydrases, the carbohydrases did not improve perfor-
mance of poultry or nutrient digestibility when they were
supplemented alone except in the studies of Selle et al.
(2003b), Wu et al. (2004a), Cowieson and Adeola (2005),
Olukosi et al. (2007, 2008) and Tiwari et al. (2010).
Synergism between supplemental phytase and carbo-
hydrases on nutrient utilization and performance can
occur if: (i) the supplemental carbohydrases are able to
hydrolyze NSP in the diet, (ii) NSP are limiting the
accessibility of phytase to PA, (iii) the PA made available
by carbohydrases is hydrolyzed by phytase, and (iv)
nutrients made available by the actions of phytase are
limiting performance. Thus, synergism between phytase
and carbohydrases is dependent on the type of supple-
mental carbohydrase, dietary NSP concentration, diet-
ary concentration of multivalent cations that affects PA
solubility, dietary concentration of non-phytate P and
endogenous phytase activity in the basal diet. All these
factors are discussed below.
VI. FACTORS THAT AFFECT INTERACTIONS
BETWEEN PHYTASE AND CARBOHYDRASES
Type of Supplemental Carbohydrases
As previously indicated, different types of NSP are
present in different feedstuffs at different concentrations.
Thus, the supplemental carbohydrases will effectively
hydrolyze NSP to increase the accessibility of phytase to
PA if their substrates are the major NSP in the diet. For
instance, the efficacy of supplemental carbohydrases in
wheat-, rye-, triticale- or corn-based diets can be higher if
they (carbohydrases) have higher xylanase activity,
because the major NSP in these feedstuffs are arabinox-
ylans. In the study of Juanpere et al. (2005), in which
there was no synergistic interaction between phytase and
carbohydrase on growth performance, AME, or reten-
tion of Ca or P of broilers fed corn-based diet, the
carbohydrase used was a-galactosidase. Thus, this could
partly explain why the authors did not observe the
interactions.
Oilseed meals, which account for approximately 50%
of PA in practical poultry diets, contain NSP whose
composition is different from those found in cereal grains
(Choct 1997). Thus, supplementation of phytase plus a
preparation containing carbohydrase enzymes that tar-
get all the major NSP in the diet (in both cereal grains
and oilseed meals), may result in improved nutrient
utilization than supplementation of phytase plus carbo-
hydrases that only target the major NSP in cereal grains
as has been done in most cases. This hypothesis is
supported by results from our recent study, which
showed additive effects of phytase and a multi-carbohy-
drase product in corn-based diets for broilers (Woyengo
et al. 2010b). It would thus, be interesting to see the effect
of supplementing a preparation containing carbohydrase
enzymes that target all the major NSP in wheat-based
diets for poultry on nutrient utilization.
Dietary Concentration of Non-starch
Polysaccharides
The efficacy of carbohydrases appears to be partly
dependent on the concentration of the target NSP (their
substrate) in the diet as indicated by results from studies
of Adeola and Bedford (2004) and Jozefiak et al. (2006).
Adeola and Bedford (2004) observed improved perfor-
mance of White Pekin ducks after xylanase supplemen-
tation of wheat-based diets when the wheat was high in
NSP (15.3%), but not when it was low in NSP content
(9.4%). Jozefiak et al. (2006) reported increased growth
performance and reduced ileal digesta viscosity of
broilers fed a rye-based diet (which was high in total
NSP 13.78%), but not wheat- and triticale-based diets,
which were lower in total NSP content (11.46 and
13.06%, respectively). Thus, a higher dietary NSP con-
centration may result in a greater response to carbohy-
drases that target them and hence synergism between
phytase and the carbohydrases.
 WOYENGO AND NYACHOTI * PHYTASE AND CARBOHYDRASES FOR POULTRY DIETS
In our broiler study (Woyengo et al. 2008) in which
synergistic interactions between phytase and xylanase on
growth performance, or Ca, P or AA digestibilities were
not observed, the wheat used to formulate the basal diet
was lower in total NSP content (8.2%) than the 10.9%
in the basal diet used in the study reported by Selle et al.
(2009) in which synergism between the two enzymes was
observed on AA digestibility in broilers. The NSP in
wheats used in the studies of Ravindran et al. (1999)
and Zyla et al. (1999a), in which synergistic interactions
between phytase and xylanase on growth performance
and AA digestibilities were observed, respectively, were
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not reported. The wheats used in these studies of
Ravindran et al. (1999) and Zyla et al. (1999a) were,
however, low in AME values (2646 and 2897 kcal kg1,
respectively), implying that they were also high in NSP
content because the AME values of the wheats were
similar to those reported by Selle et al. (2009); 2940 kcal
kg1), and AME value of wheat is negatively correlated
with NSP content (Zyla et al. 1999b; Carre et al. 2002).
Ravindran et al. (1999) did not observe any synergistic
interaction between phytase and xylanase in wheat-
based diets for broilers when the AME value for wheat
used was 3239 kcal kg1.
The NSP in wheats used in other broiler studies (Wu
et al. 2004a; Juanpere et al. 2005; Olukosi and Adeola
For personal use only.
2008) and in a layers study (Silversides et al. 2006) in
which no synergistic interactions between phytase and
xylanase were observed on nutrient utilization were also
not reported. However, the NSP in wheat used in the
study of Juanpere et al. (2005) could have been low
because the digesta viscosity for the diet used in this
study was only 5.2 cps, which is comparable with that
observed in our study (Woyengo et al. 2008; 3.65 cps) in
which we did not observe any synergistic interactions
between the two enzymes on growth performance, or
Ca, P or AA digestibilities of broilers. The NSP content
in the wheat used in the studies of Olukosi and Adeola
(2008) and Silversides et al. (2006) could also have been
low because the studies were conducted in North
America, where wheat has generally high AME content
(greater than 3200 kcal kg 1) (Bedford et al. 1998; Scott
et al. 1998a,b), implying that it has a low NSP content.
In the study of Wu et al. (2004a), supplementation of
xylanase alone improved growth performance, digesta
viscosity and weight of small intestine relative to live
body weight of broilers, implying that the wheat used
was high in NSP content. It should, however, be noted
that the phytase product used in this study of Wu et al.
(2004a) had side activities of cellulase, xylanase and b-
glucanase, which might have hydrolyzed the NSP that
the supplemental xylanase was targeting. Therefore, the
lack of synergistic interactions between phytase and
xylanase in the same study could be attributed to the
fact that the phytase product used contained NSP-
degrading enzymes. In the study of Selle et al. (2003b),
supplementation of xylanase alone also improved AA
digestibility and body weight gain of broilers, whereas
185
supplementation of a combination of phytase and
xylanase resulted in numerically higher AA digestibility
and body weight gain of broilers compared with those
fed diets supplemented with phytase or xylanase alone.
The AME values for the wheat used in the study of Selle
et al. (2003b) were fairly low (ranging from 2818 to 3184
kcal kg1, as-fed basis), and hence they may explain the
observed responses to enzyme supplementations.
From these studies, it is apparent that synergism
between phytase and xylanase in wheat-based diets for
broilers may not occur if wheat used has a total NSP
content of less than 10% or has an AME value of higher
than 3000 kcal kg 1.
The digesta viscosity for the barley-based diet used in
the study of Juanpere et al. (2005) in which synergistic
interaction between phytase and b-glucanase on P
retention in broilers was observed was high (13.7 cps).
This indicates that the barley used in this study was also
high in NSP and, hence, the reason for positive interac-
tion between the enzymes on P retention. There is,
however, limited information on the influence of NSP
or AME value of barley on the interaction between
phytase and b-glucanase because there is limited research
on the effect of supplementing a combination of phytase
and b-glucanase to barley-based diets on nutrient uti-
lization in poultry; this has only been determined in the
study of Juanpere et al. (2005). Thus, it is difficult to
determine the content of NSP in barley below which
phytase and b-glucanase are not expected to interact
synergistically with regard to nutrient utilization by
poultry.
Corn has lower NSP content than wheat or barley
(Choct 1997). Thus, the lack of synergistic interactions
between phytase and xylanase in corn-based diets used
in the broiler studies of Cowieson and Adeola (2005),
Olukosi et al. (2007, 2008), Lu et al. (2009) and Tiwari
et al. (2010) could partly be attributed to the lower
concentration of arabinoxylans (xylanase substrate) in
corn. Supplementation of xylanase alone improved
growth performance, nitrogen, energy and P digestibil-
ities of broilers in the studies of Cowieson and Adeola
(2005), Olukosi et al. (2007, 2008) and Tiwari et al.
(2010). However, the xylanase products used in these
studies also contained amylase and protease activities,
which might have improved the availability of nutrients
that were not encapsulated in cell walls because they are
not NSP-degrading enzymes. Supplementation of xyla-
nase alone did not affect performance of broilers in the
study of Lu et al. (2009) in which the xylanase product
used did not contain activities of other enzymes. Thus,
xylanase alone (as the only NSP-degrading enzyme in
the diet) may not interact synergistically with phytase in
corn-based diets for poultry.
Dietary Digestible Phosphorus Concentration
The catalytic activity of phytase is inhibited by its end
product, inorganic P (Greiner et al. 1993), indicating
that an increase in dietary concentration of non-phytate
 186 CANADIAN JOURNAL OF ANIMAL SCIENCE
P can result in decreased hydrolysis of PA by phytase.
Also, the efficiency of absorption of P from gut is
negatively correlated with dietary digestible P concen-
tration (Saddoris et al. 2010), meaning that a greater
dietary supply of non-phytate P (than required by an
animal) can result in reduced utilization of P released
from phytate by phytase. Carbohydrase and phytase can
act synergistically if phytase is able to hydrolyze PA and
if the nutrients released by phytase from PA are limiting
performance. Thus, the efficacy of phytase and hence
synergism between phytase and carbohydrases can be
limited by high dietary supply of non-phytate P.
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Several studies have shown little or no response to
phytase supplementation when the diets are formulated
to meet or exceed the available P recommendation. For
instance, Wu et al. (2003) observed greater improvement
in ileal P digestibility (by 23.7 vs. 7.4%) in broiler chicks
due to phytase when the dietary non-phytate P was
0.30% than when it was 0.45% (the dietary recom-
mended level for broiler chicks), whereas Driver et al.
(2005) reported 9.3 and 0% improvement in tibia ash of
broiler chicks due to phytase when the enzyme was
supplemented to diets with 0.24 and 0.50% non-phyate
P contents, respectively. With regard to a combination
of phytase and carbohydrases, Ravindran et al. (1999)
observed improved AA digestibility in broilers fed P-
For personal use only.
adequate diets due to supplementation of a combination
of phytase and xylanase compared with supplementa-
tion of phytase or xylanase alone, but this did not
translate into improved growth performance. Selle et al.
(2003b) reported that supplementation of a combination
of phytase and xylanase to broiler diets resulted in a
tendency for superior growth performance than that for
phytase or xylanase alone when the basal diet was
inadequate in non-phytate P (0.39%), but not when the
basal diet was adequate in non-phytate P (0.52%). Also,
the basal diets used in the study of Wu et al. (2004a), in
which there were no synergistic interactions between
phytase and carbohydrases on the response criteria
measured, the dietary level of non-phytate P was
adequate, and this could partly explain the lack of
interactions between the enzymes.
Manangi and Coon (2008) fed broiler chicks diets
containing 0.45, 0.38, 0.33, 0.28, 0.23, 0.18, 0.13 and
0.08% non-phytate P and observed a linear increase in
PA-bound P hydrolysis with the decreased dietary non-
phytate P concentration due to phytase supplementa-
tion, indicating that phytase is more effective if dietary
non-phytate P is reduced to as low level as possible.
However, the performance of broilers was not fully
restored by phytase when dietary non-phytate P was
reduced to 0.22% and below, whereas that of laying hens
was fully restored by phytase when dietary non-phytate P
was reduced to 0.10% (see Table 4). This is attributed to
the higher dietary requirement of non-phytate P by
broilers than by laying hens and the inability of supple-
mental phytase to hydrolyze enough PA-bound P to
meet the requirements. Thus, the non-phytate P content
in diets for broilers should not be reduced beyond 0.23%
if phytase is to be supplemented alone, whereas that for
laying hens can be reduced to as low as 0.10% when
phytase is to be supplemented alone.
If supplemental phytase alone can restore perfor-
mance of broilers fed basal diets whose non-phytate P
has been reduced to 0.23%, then there is a possibility
that the performance of broilers fed diets in which non-
phytate P is reduced to 0.22% or below 0.22% will
be restored by supplementation of a combination of
phytase and carbohydrases. For instance, Selle et al.
(2009) observed restoration of growth performance and
bone mineralization of broilers by supplemental phytase
alone when a basal diet with inadequate non-phytate P
content (0.26%) was fed, implying that there was an
opportunity to further reduce the dietary non-phytate P
content without an effect on growth performance and
bone mineralization of broilers if phytase and xylanase
were to be supplemented in combination. Based on
results from the studies of Olukosi et al. (2007), Tiwari
et al. (2010) and Woyengo et al. (2010b), supplementa-
tion of carbohydrases individually can improve the
digestibility of dietary P by an average of 0.09 percen-
tage points. Thus, the non-phytate P in basal diets for
broilers may be reduced to 0.14% without any affect on
performance and nutrient utilization when phytase and
carbohydrases are supplemented in a combination. For
laying hens, however, all the inorganic sources of P, such
as mono- and di-calcium phosphates, may be removed
from their diets without any affect on performance when
phytase is supplemented alone. Therefore, phytase and
carbohydrases may not act synergistically in improving
laying hens’ performance if the latter is being limited by
P availability.
Endogenous Phytase Activity
As discussed in Section II, wheat, barley, triticale and
rye, and their milling by-products have high endogenous
phytase activity. The digestibility of P in these feed
ingredients is thus higher than that in other grains like
corn. For instance, Francesch et al. (2005) observed
higher P digestibility in barley- than in corn-based diets
for laying hens (49 vs. 39%). Due to the higher
digestibility of P in diets based on wheat, barley, triticale
and rye, and their products, the impact of phytase on P
availability and utilization in diets based on these feed
ingredients is lower. Scott et al. (2001) reported im-
proved egg production and egg shell quality due to
phytase supplementation to corn-based, but not to
wheat-based diets after feeding laying hens on the basal
diets (corn- and wheat-based diets) that contained
similar calculated non-phytate P content. In our labora-
tory, we also observed a greater increase in P digestibility
(20.5 vs. 7.2 percentage units) in broilers due to phytase
when diets were based on corn (Emiola et al. 2007), but
not wheat (Woyengo et al. 2008). Similarly, Juanpere
et al. (2005) observed a positive interaction between
phytase and b-glucanase in a low-P barley-based diet
 WOYENGO AND NYACHOTI * PHYTASE AND CARBOHYDRASES FOR POULTRY DIETS
(with lower endogenous phytase activity; 257 FTU kg1)
for broilers, but no synergistic interaction between
phytase and xylanase in a low-P wheat-based diet (with
higher endogenous phytase activity; 459 FTU kg 1) for
broilers. We also did not observe any synergistic inter-
action between supplemental phytase and xylanase in a
low-P wheat-based diet (with high endogenous phytase
activity; 524 FTU kg 1) for broilers (Woyengo et al.
2008).
Some of the endogenous phytase present in feed
ingredients is inactivated by the heat generated during
feed pelleting, which is a common practice in the poultry
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feed industry, because it is sensitive to heat. For
example, Slominski et al. (2007) observed reduced
activity of phytase in wheat-based diet by 59% after
steam pelleting at 678C, whereas Afsharmanesh et al.
(2008) observed reduced activity of phytase in wheat
grain by 43% due to heat treatment at 808C. Wu et al.
(2003) and Silversides et al. (2004) reported improved
performance of broilers fed low-P wheat-based diets that
had been pelleted at 65 and 808C, respectively, due to
phytase supplementation, whereas Selle et al. (2009)
observed a positive interaction between phytase and
xylanase with regard to AA digestibility in broilers fed a
low-P wheat-based diet that had been pelleted at 708C.
The optimum temperature for plant phytases ranges
For personal use only.
from 50 to 558C (Table 2), and they lose considerable
amounts of their activities if they are subjected to
temperatures of 608C and above (Paik 2003; Sung
et al. 2005). Thus, the impact of supplemental phytase
and hence of a combination of supplemental phytase
and carbohydrases may be higher if viscous grain-based
diets are pelleted. This is because feeds are subjected to
high temperatures (65 to 808C) during the pelleting,
which can destroy the endogenous phytase, thereby
increasing the impact of supplemental phytase on
nutrient digestibility. However, there is limited informa-
tion on the effect of combining phytase and carbohy-
drases in pelleted viscous grain-based diets on nutrient
utilization in poultry, and thus, there is a need to do
more research in this area.
Dietary Concentration of Multivalent Cations
Phytic acid can react with multivalent cations at
intestinal pH to form insoluble PA-mineral complexes
that cannot be hydrolyzed by phytase (Maenz 2001).
Thus, an increase in dietary concentration of multivalent
cations can result in reduced efficacy of phytase on PA
hydrolysis and hence synergism between phytase and
carbohydrases. Of the multivalent cations that are
present in poultry diets, Zn2 is the most potent
inhibitor of PA hydrolysis followed by Fe2, Mn2,
Fe3, Ca2 and Mg2 in decreasing order of potency
(Maenz et al. 1999). However, the efficacy of phytase is
influenced more by Ca2 than the other cations,
because Ca is the major multivalent cation in practical
diets (Maenz 2001).
187
The activity of phytase was shown to be reduced in
vitro by almost 50% when Ca was added to the
incubation medium at 10% of the recommended level
for broiler chicks (Tamim et al. 2004). Phytase activity
in vivo (in broiler chicks) was reported to be reduced by
more than 60% when Ca concentration in a low-P diet
(0.13% non-phytate P) was increased from 0.2 to 0.7%
(Tamim et al. 2004). Li et al. (1999) found that a
reduction in dietary Ca level from adequate (0.8%) to
inadequate (0.4%) in the presence of microbial phytase
at 750 FTU kg1 increased the digestibility of phytate P
in piglets by 10.9 and 5.7 percentage units at the
duodenal and ileal levels, respectively. Because of this
reduced activity of phytase due to Ca, the dietary level
of Ca is often reduced by 10 to 50% when phytase is
supplemented. Manangi and Coon (2008) observed an
increase in P equivalency values for phytase in broilers
due to a reduction in dietary non-phytate P when dietary
Ca level was low (0.50%), and an increase in P
equivalency values for phytase due to an increase in
dietary non-phytate P when dietary Ca level was
adequate (0.90%). These indicate that it is critical to
also reduce dietary non-phytate P level when dietary Ca
level is reduced to maintain the same Ca to non-phytate
P ratio. Calcium to non-phytate P ratios of 2.1:1 to 2.7:1
in phytase-supplemented broiler diets have been re-
ported to be optimal (Qian et al. 1997). However, it
might be appropriate to use a basal diet with a higher
(wider) Ca to non-phytate P ratio (by increasing Ca
content and reducing non-phyate P in the diet) when a
combination phytase and carbohydrases is supplemen-
ted because higher amounts of Ca would be required
for utilization of the extra P made available due to
the addition of carbohydrases to phytase-supplemented
diets. The addition of vitamin D to the diets, which
increases the Ca absorption (Bronner 1987), or organic
acids such as citric acid, which increase the solubility of
Ca-PA complexes (Boling et al. 2000), may alleviate the
negative effects of Ca on PA hydrolysis by phytase and
hence result in improved performance and bone miner-
alization. However, this hypothesis needs to be tested.
VII. CONCLUSIONS
Synergism between phytase and carbohydrase in poultry
diets depends on several factors including the type of
carbohydrase product used, the composition and con-
tent of dietary NSP, the dietary concentration of non-
phytate P and Ca, and the endogenous phytase activity
in the diets used. The synergistic effects are more
pronounced in diets that use carbohydrase product
with enzyme activities that target most of the major
NSP in the basal diet. Supplemental xylanase can
interact synergistically with phytase in wheat-based diets
for poultry only if the wheat used has a higher
concentration of NSP (greater than 10%). The utiliza-
tion of phytase in broiler diets can be improved by
carbohydrases if the non-phytate P in the basal diet is
reduced by a bigger margin (to around 0.14%) than is
 188 CANADIAN JOURNAL OF ANIMAL SCIENCE
often used (to a range of 0.23 to 30%) when phytase is
supplemented alone. However, there may be no need to
reduce the dietary Ca level to keep a narrow Ca to non-
phytate P ratio as is often done when phytase is
supplemented alone, because a greater amount of Ca
would be required for the utilization of the extra P made
available due to the addition of carbohydrases to
phytase-supplemented diets. Phytase and carbohydrases
may not interact synergistically or additively in poultry
diets if the basal diet has high endogenous phytase
activity. There is, however, a need to establish the endo-
genous phytase activity level beyond which phytase and
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