Effects of residual superdoses of phytase on growth performance, tibia

mineralization, and relative organ weight in ducks fed phosphorus-deficient

diets

L. Fan,∗,1 Z. Z. He,†,1 X. Ao,‡ W. L. Sun,∗ X. Xiao,∗ F. K. Zeng,∗ Y. C. Wang,∗ and J. He∗,2

∗
College of life science and engineering, Southwest University of Science and Technology, Mianyang, Sichuan,
621010, P. R. China; † Faculty of Animal science, University of Adelaide, Adelaide, 5005, Australia; and ‡ Tie Qi
Li Shi Group. Co., Mianyang, Sichuan, 621006, P. R. China

ABSTRACT This study was conducted to determine
the effects of residual superdoses of phytase on growth
performance, tibia mineralization, and relative organ
weight in ducks fed phosphorus-deficient diets. In Exp.
1, 4 kinds of commercial phytase were used to determine
retention rate of phyatse with the phytase C being the
highest via both high water-bath temperature (90%)
and pelleting (50%), followed by phytase A, B, and
D. In Exp. 2, a total of 560 male ducks were blocked
based on body weight, and then allocated randomly to
7 treatments (5 replicates with 16 birds per replicate).
Treatments included a maize-soybean meal-based diet
with recommended calcium and 4.0 g non-phytate phos-
phorus (nPP)/kg starter diet or 3.8 g nPP/kg grower
diet (positive control; PC), an nPP-deficient diet with
1.3 g nPP/kg starter diet or 1.1 g nPP/kg grower diet
(negative control; NC), NC diets with increasing levels
of residual phytase C (500, 1,000, 2,000, 3,000, and
4,000 units/kg feed) after pelleting. Birds fed NC diets
Key words: ducks, phytase, performance, tibia mineralization
had lower (P < 0.05) average daily gain (ADG) and
average daily feed intake (ADFI) throughout the exper-
iment compared with those fed PC diet. Supplementing
NC diet with increasing residual superdoses of phytase
improved (P < 0.05) ADG and ADFI quadratically in
the entire experiment, while reduced feed-to-gain ratio
(P < 0.05) quadratically during day 0 to 14. On day
14 and 35, birds fed NC diet had lower (P < 0.05)
tibia length, weight, ash, calcium, phosphorus, and
manganese contents than those fed PC diet. Increasing
residual superdoses of phytase in NC diet increased
(P < 0.05) tibia weight and ash, calcium, phosphorus
contents quadratically on day 14 and 35. NC treatment
increased (P < 0.05) the duodenum, jejunum, ileum,
and cecum index compared with other treatments on
day 14 and 35. Taken together, feeding increasing resid-
ual superdoses of phytase could counteract or exceed
the negative effects of NC diet on growth performance,
tibia mineralization, and relative organ weight in ducks.
2019 Poultry Science 98:3926–3936
http://dx.doi.org/10.3382/ps/pez114

INTRODUCTION thermore, several studies have previously demonstrated

Phosphorus (P) is an essential macromineral in-
volved in normal growth, bone formation, and a variety
of metabolic pathways (NRC, 1994). However, it is
well documented that about 60 to 70% P in cereal
grains (Nelson, 1967) and 35 to 50% P in oilseeds such
as soybean meal and rapeseed cake (Eeckhout and De
Paepe, 1994) are in the form of phytate (Cheryan, 1980;
Harland and Oberleas, 1999; Adeola and Sands, 2003).
Phytate, the salt of phytic acid (myo-inositol hexak-
isphosphate, InsP6), is unavailable to poultry due to the
lack of the necessary endogenous enzymes to properly
hydrolyze phytic acid (Adeola and Sands, 2003). Fur-

C 2019 Poultry Science Association Inc.
Received November 11, 2018.
Accepted February 26, 2019.
1
These authors contributed equally to this work.
2
Corresponding author: hjsc646@aliyun.com
that phytate may bind nutrients (such as calcium,
zinc, ferrum, magnesium, copper, starch, lipids, and
certain amino acids), which makes them less available
to poultry (Viveros et al., 2002; Cowieson et al., 2006;
Onyango and Adeola, 2009; Pirgozliev et al., 2012).
The low non-phytate phosphorus (nPP) diets in
ducks have negative influence on growth performance
and mineral utilization. Ducks fed low nPP diets have
been shown to decrease final body weight (BW), feed
intake, nutrient utilization, and bone mineralization
(Ravindran et al., 1995; Rodehutscord et al., 2003; Dai
et al., 2018) as well as influence the cecal microbiota
(Dai et al., 2018). Due to the rapid genetics progress
in ducks during recent decades, growth performance
has been improved greatly, which also may cause
serious health issues, such as leg problems, abnormal
bone, and skeletal development (Adeola, 2010, 2018;
Zeng et al., 2015). Williams et al. (2000) indicated
that selected fast-growing strains have shown lower

3926
 RESIDUAL SUPERDOSES OF PHYTASE IN DUCKS
Table 1. The determination of 4 kinds of commercial microbial phytase.
Determined activity,
Items Moisture, % units/g
A 3.23 10,777
B 0.67 11,714
C 7.16 10,837
D 9.32 11,665
bone-ash content than slow-growing strains. Therefore,
it is suggested that ducks diets should be higher in Ca
and P than current recommendations in order to reach
skeletal integrity for modern strains.
The use of phytase in low nPP diets benefits
phytate hydrolysis (Orban et al., 1999), and has been
demonstrated to improve the availability of P, BW
gain, bone mineralization, and nutrient utilization in
ducks (Orban et al., 1999; Rodehutscord et al., 2006;
Adeola, 2010). These benefits have been observed in
a range of doses from 500 to 2,000 units/kg of phytase
in diets, with the industry using a uniform inclusion
level around 500 units/kg mark. With the development
of technology, the production cost of phytase has
decreased greatly, which makes it cost-effective to
include superdoses by the industry. In a recent study,
superdoses (15,000 units/kg) of phytase improved BW
gain, feed intake, feed efficiency, ileal digestibility, and
retention of P in 43-day-old ducks fed diets with low
nPP (Adeola, 2018), which was consistent with the
findings in broilers fed diets with superdoses of phytase
(Walk et al., 2013, 2014; Taheri and Taherkhani,
2015; Farhadi et al., 2017; Pieniazek et al., 2017). The
improvements may be attributed to nearly complete
hydrolysis of phytate and the removal of antinutritive
effects (Shirley and Edwards, 2003), higher nutrient
digestion and feed efficiency (Dos Santos et al., 2013;
Walk et al., 2013, 2014), increased immunity and an-
tioxidant status (Liu et al., 2008, 2010a; Karadas et al.,
2010), altered gene expression associated with growth
(Liu et al., 2010b; Woyengo et al., 2011), and decreased
excreta moisture content (Delezie et al., 2015).
It is supposed that the inclusion level of above 1,500
units/kg of phytase is defined as superdoses. Due to
the aforementioned achievements in the phytase appli-
cation, the potential of superdoses of phytase (>1,500
units/kg) has gained scientific and industrial interest.
Notwithstanding, little information about the super-
doses of phytase in ducks was available. Ducks differ in
P digestion from other poultry species (Rodehutscord
and Dieckmann, 2005). Different types of phytase are
denatured to different extents when subjected to the
temperature and moisture of preconditioning during
pelleting. However, the previous studies analyzed the
phytase activity before pelleting. Undoubtedly, the re-
search about the effect of superdoses of phytase on
ducks, especially in the form of residual phytase activ-
ity after pelleting, would be of great value. Therefore,
3927
Strains of bacteria Color Coated Form
Aspergillus oryzae White with Yes Fine granule
little yellow
Aspergillus oryzae White Yes Fine granule
Aspergillus niger White No Powder
Aspergillus niger White Yes Fine granule
the objective of this study was to further investigate the
influences of residual superdoses of phytase on growth
performance, tibia mineralization, and relative organ
weight in ducks fed P-deficient diets.
MATERIALS AND METHODS
Experimental Design and Duck Husbandry
In Exp. 1, 4 kinds of commercial microbial phytase
(A from Aspergillus oryzae, B from A. oryzae, C from
Aspergillus niger, and D from A. niger), all of which are
expressed in Pichia pastoris and guaranteed to contain
10,000 units/g, are selected for the determination of
phytase activity. One unit of phytase activity is defined
as the quantity of enzyme required to hydrolyze 1 μmol
of inorganic P/min, at pH 5.5, from an excess of 5.0 mM
sodium phytate at 37◦ C (Engelen et al., 1994). Then,
4 kinds of commercial microbial phytase were treated
by high water-bath temperature and pelleting. The 4
kinds of commercial microbial phytase were treated by
the water bath at 85◦ C for 3 and 4 min, and then cooled
in ice water at 0◦ C for 30 min to determine the residual
phytase activity. The negative control (NC) starter diet
with 1 g phytase A, B, C, and D/kg feed was pelleted
for the determination of retention rate of the phytase
activity. Mash feed was steam conditioned for 30 s and
pelleted at 85◦ C, with temperatures monitored within
the conditioner and as the feed exited the pelleter.
In Exp. 2, the Animal Welfare Committee of South-
west University of Science and Technology approved the
animal care protocol used for this experiment. A total
of 560 one-day-old male Cherry Valley ducks with an
average initial BW of 55.1 ± 0.2 g were blocked on
the basis of BW, and placed in stainless steel battery
brooders (2.0 × 1.0 m). All ducks were housed in an
environmentally controlled facility.
This 5-wk experiment consisted of 7 treatments with
5 pens per treatment and 16 ducks per pen in a random-
ized complete block design. A 2-phase feeding program
was used: a starter diet from day 1 to 14 and a grower
diet from day 15 to 35. As presented in Table 4, all diets
were formulated to meet or exceed the duck’s nutri-
tional requirements (NRC, 1994) with the exception of
nPP levels. Dietary treatments included a corn-soybean
meal-based diet with recommended levels of nPP (4.0 g
nPP/kg starter or 3.8 g nPP/kg grower diet) as
positive control (PC); an nPP-deficient corn-soybean
3928 FAN ET AL.
meal-based diet (1.3 g nPP/kg starter or 1.1 g nPP/kg
grower diet) as NC; NC diets which were supplemented
with graded levels of residual phytase (500, 1,000, 2,000,
3,000, and 4,000 units/kg feed). Expected residual phy-
tase activities were confirmed by analysis (Table 5). To
achieve maximize homogeneity of added phytase with
diet, at first a small quantity of each complete diet was
mixed with the enzyme and then mixed with whole of
the diet to get the final concentration. Diets were fed in
pellet form and feed and water were provided ad libitum
throughout the experiment.
Proximate analysis of diets for dry matter (DM) and
ash was carried out according to AOAC (2000). The
DM of the feed was determined after drying for 24 h at
103◦ C. Ash was determined after ignition of a weighed
sample in a muffle furnace (Nabertherm, Bremen, Ger-
many) at 550◦ C for 6 h. The ash was then digested in
aqua regia (HCl/HNO3 mixture), and the solution was
used for calcium and P determination. Calcium con-
centration was determined using an atomic absorption
spectrophotometer (Varian’50, Varian, Palo Alto, CA),
and the concentration of P was analyzed spectrophoto-
metrically (NanoDrop 2000c, Thermo Scientific, MA)
using the method of Cavell (1995).
Sampling and Measurements
The ducks were weighed and feed intake was recorded
on day 1, 14, and 35, and average daily gain (ADG),
average daily feed intake (ADFI), and feed-to-gain ra-
tio (F/G) were calculated.
At the end of the second week and the experiment,
4 ducks were randomly selected from each pen and the
ducks were sacrificed by cervical dislocation. The heart,
liver, spleen, lung, bursa fabricius, left tibia, duodenum,
jejunum, ileum, and cecum were removed by trained
personnel. The heart, liver, spleen, lung, bursa fabri-
cius, and left tibia were weighed after flushing with
saline. Organ size was expressed as a percentage of BW.
The duodenum, jejunum, ileum, and cecum index was
expressed as a length percentage of BW.
The left tibia bones were removed, defleshed from
adherent soft tissues, and then stored at –20◦ C until
further analysis (length and mineral determination).
The left tibia bones were dehydrated in ethanol for
72 h, defatted for 72 h in diethyl ether: methanol (9:
1), and dried at 105◦ C for 24 h to constant weight.
Then, tibia bones were ashed in a muffle furnace at
600◦ C overnight in porcelain crucibles and the tibia
ash percentage was expressed as grams of ash per
100 g of dry, fat-free weight of the tibia. The con-
centrations of Ca, Zn, Cu, Fe, Mn, and Mg in the
tibia were determined using an atomic absorption spec-
trophotometer (Varian’50) according to AOAC (2000),
and the concentration of P was determined spectropho-
tometrically (NanoDrop 2000c) using the method of
Cavell (1995).
Statistical Analysis
In Exp. 1, data were analyzed by ANOVA using the
GLM procedure of SAS (SAS Inst. Inc., Cary, NC). Sig-
nificant differences among all dietary treatments were
determined at P < 0.05 by Tukey tests. Variability in
the data is expressed as the standard error of means
(SEM). Quadratic regression analysis was performed
for the residual phytase activity correlation between
high water-bath temperature and pellet.
In Exp. 2, data were analyzed by ANOVA using
the GLM procedure of SAS (SAS Inst. Inc.) with the
pen being the experimental unit. Significant differences
among all dietary treatments were determined at P <
0.05 by Tukey tests. Variability in the data is expressed
as SEM. The dose–response effect of supplemental phy-
tase was computed using orthogonal polynomial con-
trast for liner and quadratic effects. The optimal re-
quirement of residual superdose was analyzed with the
broken quadratic model.
RESULTS
Determination of Commercial Microbial
Phytase
In Exp. 1, the product information about 4 kinds of
commercial phytase (A, B, C, and D), which are guar-
anteed to contain 10,000 units/g, is shown in Table 1.
All of the determined phytase activities were greater
than 10,000 units/g. The retention rate of 4 kinds of
commercial phytase was significantly different (P <
0.05) with C being the highest (88 to 92%), followed by
A (83 to 88%), B (41 to 42%), and D (1 to 5%) by high
water-bath temperature for 3 and 4 min (Table 2). After
pelleting, the retention rate of C (50%) and A (49%)
was higher (P < 0.05) than that of B (44%) and D
(17%). The high water-bath temperature and time were
significantly correlated with pelleting treatment (P <
0.05). The results of regression equation are as follows:
Retention rate of pelleting
= a ∗ retention rate of high water-bath temperature2
+ b ∗ retention rate of high water-bath temperature
+ c (Table 3).
In Exp. 2, the determined residual activity of phytase
in the diets was in agreement with formulated values
after pelleting (Table 5).
Growth Performance
During day 0 to 14, birds fed NC diet with low
nPP level (2.7 g nPP/kg diet less nPP vs. PC) had
lower ADG and ADFI (P < 0.05), but higher F/G
(P < 0.05; Table 6). During day 15 to 35, birds fed
NC diet had lower (P < 0.05) ADG, ADFI, and F/G
than those fed PC diet. In the entire experiment, ADG
and ADFI in NC treatment were reduced (P < 0.05)
 RESIDUAL SUPERDOSES OF PHYTASE IN DUCKS 3929
Table 2. Retention rate of 4 kinds of commercial microbial phytase activity in high water-bath temperature
and the pelleted NC starter diets.

Retention rate, %
Items1 A B C D SEM P-value

Treated for 3 min 88.56a 42.41b 92.82a 5.65c 0.48 < 0.01
Treated for 4 min 83.66b 41.09c 88.00a 1.58d 0.50 < 0.01
Pelleted 49.99a 44.68b 50.24a 17.50c 1.01 < 0.01
1
Treated for 3 min, the 4 kinds of commercial microbial phytase were treated by the water bath at 85◦ C for 3 min,
and then cooled in ice water at 0◦ C for 30 min to determine the residual phytase activity; treated for 3 min, the 4 kinds
of commercial microbial phytase were treated by the water bath at 85◦ C for 4 min, and then cooled in ice water at 0◦ C
for 30 min to determine the residual phytase activity; pelleted, negative control maize-soybean meal-based starter diets
with low nPP content and 10,000 phytase A, B, C, and D units/kg feed, respectively, were pelleted.

Table 3. Parameters of quadratic regression analysis between high water-bath temperature and
pellet.1

Items2 a b c R2(3) P-value

Treated for 3 min –0.007 1.084 11.644 0.911 < 0.01

Treated for 4 min –0.007 0.968 16.013 0.925 < 0.01
Pelleted 0.000 1.000 0.000 1.000 < 0.01
1
Treated for 3 min, the 4 kinds of commercial microbial phytase were treated by the water bath at 85◦ C
for 3 min, and then cooled in ice water at 0◦ C for 30 min to determine the residual phytase activity; treated
for 3 min, the 4 kinds of commercial microbial phytase were treated by the water bath at 85◦ C for 4 min, and
then cooled in ice water at 0◦ C for 30 min to determine the residual phytase activity; pelleted, negative control
maize-soybean meal-based starter diets with low nPP content and 10,000 phytase A, B, C, and D units/kg
feed, respectively, were pelleted.
2
Regression equation: retention rate of pelleting = a ∗ retention rate of high water-bath temperature2 + b
∗
retention rate of high water-bath temperature + c.
3
Pearson correlation coefficient between high water-bath temperature and pellet.

compared with PC treatment. Supplementation of NC
diet with increasing residual superdoses of phytase im-
proved (P < 0.05) ADG and ADFI quadratically from
day 0 to 14, day 15 to 35, and day 0 to 35. In ad-
dition, F/G was decreased (P < 0.05) quadratically
with increasing residual superdoses of phytase during
day 0 to 14.
Tibia Mineralization
On day 14 and 35, the tibia length and weight in
NC treatment were lower (P < 0.05) than those in PC
treatment (Table 7). Addition of increasing residual su-
perdoses of phytase in NC diet increased (P < 0.05)
tibia length and weight quadratically on day 14, but
only increased (P < 0.05) tibia weight quadratically on
day 35.
On day 14, birds fed NC diet had lower (P < 0.05)
tibia ash, Ca, P, Cu, Mn, and Mg, but higher (P < 0.05)
Fe and Zn contents than those fed PC diet (Table 8).
Besides, NC treatment reduced (P < 0.05) tibia ash,
Ca, P, Zn, and Mn contents, while increased Fe content
(P < 0.05) without effect on Cu or Mg on day 35 com-
pared with PC treatment. Supplementation of NC diet
with increasing residual superdoses of phytase increased
(P < 0.05) tibia ash, Ca, P, and Mg contents quadrati-
cally, but decreased (P < 0.05) Fe content quadratically
on day 14. On day 35, addition of increasing residual
superdoses of phytase in NC diet increased tibia P, Zn,
and Mg quadratically as well as Ca linearly.
Relative Organ Weight
The duodenum, jejunum, ileum, and cecum index
as a length percentage of BW in NC treatment was
greater (P < 0.05) than those in other treatments on
day 14 and 35 (Table 9). Birds fed NC diet had higher
(P < 0.05) relative weight of heart and liver on day 14
and 35, but lower (P < 0.05) relative weight of bursa
fabricius on day 14 and lung on day 35 than those fed
PC diet (Table 10). Dietary treatment did not affect
(P > 0.05) relative weight of spleen and lung on day 14
or spleen and bursa fabricius on day 35. The relative
weight of heart in NC treatment with residual phytase
500 units/kg diets was higher (P < 0.05) than that in
PC treatment on day 14. There was no difference (P >
0.05) in relative weight of liver and bursa fabricius on
day 14 or heart, liver, and lung on day 35 between PC
and NC treatments with residual superdoses of phytase.
Optimal Requirement
Parameters of fitness between selected index and
phytase retention via quadratic broken line model
were shown in Table 11. With the addition of mea-
surement as the abscissa (x) and the evaluation index
3930 FAN ET AL.

Table 4. Diet composition (as-fed basis).

Starter1 Grower1
2 2 2
Item, % NC PC NC PC2

Maize 520.0 520.0 656.0 656.0

Soybean meal (CP 43%) 260.0 260.0 57.5 57.5
Rice bran meal 50.0 50.0 50.0 50.0
Soybean oil 6.0 6.0 12.0 12.0
DDGS 60.0 60.0 60.0 60.0
Cottonseed meal (CP 42%) 50.0 50.0 107.0 107.0
Dialcium phosphate — 16.7 — 15.8
Limestone 12.7 7.6 13.8 10.3
Bentonite 21.2 9.6 2.18 9.5
Salt 2.0 2.0 3.0 3.0
Sodium bicarbonate 2.0 2.0 2.0 2.0
Choline chloride (50%) 1.5 1.5 1.2 1.2
DL-Met (99%) 2.7 2.7 2.2 2.2
L-Lys∼ =SO4 (70%) 5.6 1.5 8.6 8.6
L-Threonine 1.5 1.5 1.7 1.7
L-Tryptophan (20%) 1.8 1.8 0.2 0.2
Vitamin premix3 1.5 1.5 1.5 1.5
Trace mineral premix4 1.5 1.5 1.5 1.5
Analytical composition
ME, kcal/kg5 2700 2700 2900 2900
Crude protein, g/kg 212.0 214.0 161.0 162.0
Lys, g/kg 12.6 12.6 10.6 10.6
Met+Cys, g/kg 9.0 9.0 7.3 7.3
Thr, g/kg 8.6 8.6 6.7 6.7
Ca, g/kg 6.9 8.5 6.8 8.4
Total P, g/kg 5.0 7.7 4.3 7.1
Non-phytate P, g/kg5 1.3 4.0 1.1 3.8
1
Provided starter diets during week 0 to 2; provided grower diets during week 3 to 5.
2
NC = negative control diet with 2.5 and 2.3 g non-phytate P during starter and grower, respectively;
PC = positive control diet with 4.0 and 3.8 g non-phytate P during starter and grower, respectively.
3
Provided per kilogram of diet: vitamin A, 2,500 IU; vitamin D3 ,400 IU; vitamin E, 10 IU; vitamin K3 ,
0.5 mg; vitamin B1 , 2.0 mg; vitamin B6 , 2.5 mg; vitamin B12 , 0.02 mg; nicotinic acid, 55 mg; pantothenic acid,
10 mg; folic acid, 1.0 mg; and biotin, 0.1 mg.
4
Provided per kilogram of diet: 60 mg Fe (FeSO4 ·7H2 O); 8 mg Cu (CuSO4 ·5H2 O); 60 mg Zn (ZnSO4 ·7H2 O);
50 mg Mn (MnSO4 ·H2 O); 0.1 mg Se (Na2 SeO3 ·5H2 O); and 0.2 mg I (KI).
5
Calculated values.

Table 5. Expected and analyzed residual phytase activity in the According to the above model, the optimal require-
pelleted diets. ment of residual phytase superdose ranged from 555
Expected phytase Analyzed phytase
to 3,333 units/kg diet for all traits with the best
Items1 activity, units/kg activity, units/kg supplementation levels around 1,306 to 1,777 units/kg
diet for growth performance.
D 0–14
PC 0 0
NC 0 0
NC + 500 phytase units/kg 500 517
NC + 1,000 phytase units/kg 1,000 945
DISCUSSION
NC + 2,000 phytase units/kg 2,000 1,932
NC + 3,000 phytase units/kg 3,000 3,001 Determination of Commercial Microbial
NC + 4,000 phytase units/kg 4,000 3,961
Day 15 to 35
Phytase
PC 0 0
NC 0 0 There were significant quality differences in these 4
NC + 500 phytase units/kg 500 558 kinds of commercial phytase (A, B, C, and D). The de-
NC + 1,000 phytase units/kg 1,000 986 termined phytase activities also confirmed that all of
NC + 2,000 phytase units/kg 2,000 1,933
NC + 3,000 phytase units/kg 3,000 2,990 them were guaranteed to contain 10,000 units/g. How-
NC + 4,000 phytase units/kg 4,000 3,972 ever, after high water-bath temperature treatment for
1
PC, positive control maize-soybean meal-based diet with adequate
3 min, the retention rates of C and A came close to
Ca and nPP content; NC, negative control maize-soybean meal-based 90% and B was 40% with D being the lowest (nearly
diet with low nPP content; treatments with residual phytase activity of 5%). Extending the high water-bath temperature time
0, 500, 1,000, 2,000, 3,000, and 4,000 units/kg feed.
to 4 min only decreased the retention rate slightly com-
pared with that for 3 min, which may indicate that
as the ordinate (y), the quadratic broken line fitting high water-bath temperature treatment for 3 min may
equation is as follows: when x < optimal dose, y = be proper for evaluation. After pelleting, the similar
rate constant ∗ (optimal requirement- x)2 + maximum trend was observed in retention rate of 4 kinds of com-
value; when x > optimal dose, y = maximum value. mercial pyhtase. Notwithstanding, the retention rate of
 RESIDUAL SUPERDOSES OF PHYTASE IN DUCKS 3931
Table 6. Effects of superdoses of phytase on growth performance in ducks fed phosphorus-deficient diets.1

Residual phytase activity, units/kg P-value4

2 2 3
Items PC NC 500 1,000 2,000 3,000 4,000 SEM Treatment Linear Quadratic

Day 0 to 14
ADFI, g 62.9a 12.6d 45.3c 56.9b 59.6a,b 60.2a,b 60.6a,b 2.88 < 0.01 < 0.01 < 0.01
ADG, g 43.7a 7.4d 31.3c 38.8b 42.0a 43.6a 44.0a 2.14 < 0.01 < 0.01 < 0.01
F/G 1.44b 1.71a 1.45b 1.47b 1.42b,c 1.38c 1.38c 0.02 < 0.01 < 0.01 < 0.01
Day 15 to 35
ADFI, g 215.7a 122.0e 173.0d 197.9b,c 198.7b,c 209.2a 201.3b 3.59 < 0.01 0.001 < 0.01
ADG, g 104.9b,c 63.6e 93.6d 105.4b,c 107.2a,b 110.7a 110.2a 2.06 < 0.01 < 0.01 < 0.01
F/G 2.06a 1.92b 1.85b,c 1.88b,c 1.85b,c 1.89b,c 1.83c 0.01 < 0.01 0.62 0.35
Day 0 to 35
ADFI, g 148.4a 75.7e 117.0d 135.9c 137.4c 143.6a,b 139.3b 4.65 < 0.01 < 0.01 < 0.01
ADG, g 77.4b 39.8e 66.0d 75.8b,c 78.0a,b 80.7a 80.6a 2.38 < 0.01 < 0.01 < 0.01
F/G 1.92a 1.90a 1.77b 1.79b 1.76b 1.78b 1.73b 0.02 < 0.01 0.09 0.15
1
Means represent 5 pens of ducks with 16 birds per pen (n = 5/group).
2
PC, positive control maize-soybean meal-based diet with adequate Ca and nPP content; NC, negative control maize-soybean meal-based diet with
low nPP content.
3
Pooled standard error of the means.
4
Treatment, treatment effect; linear, linear effect for treatment with residual phytase activity of 0, 500, 1,000, 2,000, 3,000, and 4,000 units/kg;
quadratic, quadratic effect for treatment with residual phytase activity of 0, 500, 1,000, 2,000, 3,000, and 4,000 units/kg.
a–c
Means in the same row with different superscripts differ (P < 0.05).

Table 7. Effects of superdoses of phytase on tibia development in ducks fed phosphorus-deficient diets.1

Residual phytase activity, units/kg P-value4

2 2 3
Items PC NC 500 1,000 2,000 3,000 4,000 SEM Treatment Linear Quadratic

Day 14
Length, mm 84.1a 58.4c 78.0b 81.9a,b 84.1a 85.3a 84.1a 1.65 < 0.01 0.02 0.01
Weight, g5 1.75a,b 0.67c 1.40b 1.50b 1.78a,b 1.85a 1.86a 0.08 < 0.01 < 0.01 < 0.01
Day 35
Length, mm 119.6a 99.0b 115.7a 116.0a 116.3a 116.9a 118.6a 1.29 < 0.01 0.19 0.43
Weight, g 5.35a 3.13c 4.43b 5.04a,b 5.17a,b 5.21a,b 5.02a,b 0.13 < 0.01 < 0.01 < 0.01
1
Means represent 5 pens of ducks with 16 birds per pen (n = 5/group).
2
PC, positive control maize-soybean meal-based diet with adequate Ca and nPP content; NC, negative control maize-soybean meal-based diet with
low nPP content.
3
Pooled standard error of the means.
4
Treatment, treatment effect; linear, linear effect for treatment with residual phytase activity of 0, 500, 1,000, 2,000, 3,000, and 4,000 units/kg;
quadratic, quadratic effect for treatment with residual phytase activity of 0, 500, 1,000, 2,000, 3,000, and 4,000 units/kg.
5
Organ size was expressed as a percentage of BW.
a–c
Means in the same row with different superscripts differ (P < 0.05).

every phytase reduced even more sharply after pellet-
ing compared with high water-bath temperature treat-
ment. Similarly, the retention rates of C (50%) and A
(50%) were higher than that of B (44%) and D (17%).
Based on the above results, C was chosen to be used
in the next animal experiment for further study about
residual phytase superdose in ducks. Besides, the re-
gression equation between the retention rate of pel-
leting and high water-bath temperature treatment
for 3 min was established to evaluate the phytase
quality:
Retention rate of pelleting = a ∗ retention rate of
high water-bath temperature2 + b ∗ retention rate of
high water-bath temperature + c.
Correspondingly, the determined residual activity of
phytase (C) was similar to the expected values after
pelleting in Exp. 2.
Growth Performance
Dietary P is an essential nutrient for normal growth,
skeletal system development, maintenance (Shastak
and Rodehutscord, 2013), and a variety of metabolic
processes in bacterial cell (Durand and Komisarczuk,
1988). The nPP requirements used in the present study
were based on the NRC (1994) requirement for duck.
The actual nPP requirement of ducks was 4 g nPP/kg
diet for 1 to 14 D of age and 3 g nPP/kg diet for 15 to
56 D of age (NRC, 1994). As expected, a lower level of
nPP (1.3 g nPP/kg diet) in the diet decreased ADG and
ADFI by 48 and 49% throughout the experiment, which
was in agreement with recent studies (Zeng et al., 2015;
Dai et al., 2018). Feeding 1.3 g nPP/kg diet, which was
the same nPP level in our study, reduced BW gain and
feed intake by 49 and 37% in ducks from the age of day
7 to 17 (Adeola, 2010). Dai et al. (2018) reported that
2.2 g nPP/kg diet reduced ADG and ADFI by 13 and
11% in ducks from day 1 to 21. Zeng et al. (2015) also
indicated a lower BW at the age of day 14 and ADFI
3932 FAN ET AL.

Table 8. Effects of superdoses of phytase on tibia mineralization in ducks fed phosphorus-deficient diets.1

Residual phytase activity, units/kg P-value4

2 2 3
Items PC NC 500 1,000 2,000 3,000 4,000 SEM Treatment Linear Quadratic

Day 14
Ash, % 47.9a 32.4d 38.1c 43.0b 44.7a,b 46.2a,b 45.7a,b 1.00 < 0.01 < 0.01 < 0.01
Ca, % 19.0a 12.8d 14.9b,c 15.6b 14.0c 18.0a,b 17.4a,b 0.49 < 0.01 0.03 0.08
P, % 8.76a 5.70d 7.38c 7.87b,c 8.03b 8.37a,b 8.32a,b 0.18 < 0.01 < 0.01 < 0.01
Cu, mg/kg 3.94a,b 3.28c 3.90a,b 3.58b,c 3.76b 4.18a 3.98a,b 0.07 < 0.01 0.06 0.17
Fe, mg/kg 128b,c 183a 146b 136b,c 122c 121c 118c 4.44 < 0.01 < 0.01 < 0.01
Zn, mg/kg 247b 278a 247b 248b 249b 225c 246b 3.81 < 0.01 0.36 0.44
Mn, mg/kg 12.6b 10.9c,d 11.3c 13.4a 10.3d 12.2b,c 12.8a,b 0.24 < 0.01 0.59 0.44
Mg, mg/kg 3610a 2210d 2550c 3090b 3403a,b 3476a 3462a 93.8 < 0.01 < 0.01 < 0.01
Day 35
Ash, % 53.9a,b 50.2c 51.4b,c 52.9b 53.9a,b 54.0a,b 55.3a 0.42 0.06 0.15 0.06
Ca, % 21.8a 20.2c 20.3c 20.9b 21.2a,b 20.8b 21.9a 0.21 0.18 0.02 0.07
P, % 10.5a 9.1b 9.3b 10.1a 10.3a 10.3a 10.6a 0.12 < 0.01 < 0.01 < 0.01
Cu, mg/kg 3.98c 3.85c 3.78c 4.36a,b 4.70a 4.28b 4.26b 0.10 0.15 0.40 0.14
Fe, mg/kg 113b 127a 111b 123a 98c 111b 107b 1.94 < 0.01 0.16 0.32
Zn, mg/kg 228a 196c 204c 239a 245a 229b 239a 3.67 < 0.01 0.04 0.02
Mn, mg/kg 14.5a 9.50b 13.6a 13.4a 13.6a 14.0a 14.7a 0.31 < 0.01 0.13 0.26
Mg, mg/kg 3429a,b 3581a 2824c 3054b 3488a 3474a 3648a 68.2 < 0.01 < 0.01 < 0.01
1
Means represent 5 pens of ducks with 16 birds per pen (n = 5/group).
2
PC, positive control maize-soybean meal-based diet with adequate Ca and nPP content; NC, negative control maize-soybean meal-based diet with
low nPP content.
3
Pooled standard error of the means.
4
Treatment, treatment effect; linear, linear effect for treatment with residual phytase activity of 0, 500, 1,000, 2,000, 3,000, and 4,000 units/kg;
quadratic, quadratic effect for treatment with residual phytase activity of 0, 500, 1,000, 2,000, 3,000, and 4,000 units/kg.
a–c
Means in the same row with different superscripts differ (P < 0.05).

Table 9. Effects of superdoses of phytase on intestinal tract index in ducks fed phosphorus-deficient diets.1

Residual phytase activity, units/kg

Items4 PC2 NC2 500 1,000 2,000 3,000 4,000 SEM3 P-value

Day 14
Duodenum 34.8b 116a 41.7b 37.9b 35.0b 35.2b 33.8b 4.94 < 0.01
Jejunum 84.9b 262.2a 103.5b 92.6b 87.6b 84.0b 84.2b 10.7 < 0.01
Ileum 84.7b 258.2a 106.3b 91.9b 89.8b 86.3b 82.6b 10.3 < 0.01
Cecum 16.5b 56.7a 20.1b 19.1b 18.9b 17.8b 17.1b 2.40 < 0.01
Day 35
Duodenum 12.0b 36.4a 13.2b 11.8b 11.7b 11.9b 11.3b 1.52 < 0.01
Jejunum 29.2b 90.5a 32.8b 29.7b 29.5b 28.2b 27.9b 3.95 < 0.01
Ileum 30.2b 91.7a 34.3b 29.9b 29.5b 29.0b 28.0b 3.99 < 0.01
Cecum 6.90b 20.7a 7.71b 6.40b 6.72b 6.11b 6.42b 0.83 < 0.01
1
Means represent 5 pens of ducks with 16 birds per pen (n = 5/group).
2
PC, positive control maize-soybean meal-based diet with adequate Ca and nPP content; NC, negative control maize-soybean
meal-based diet with low nPP content.
3
Pooled standard error of the means.
4
The duodenum, jejunum, ileum, and cecum index was expressed as a length percentage of BW.
a,b
Means in the same row with different superscripts differ (P < 0.05).

during day 15 to 35 in ducks fed 2.3 g nPP/kg diet com-
pared with those fed 2.8 g nPP/kg diet. Feeding the low
nPP diet (3.0 g nPP/kg starter diet and 2.0 g nPP/kg
grower diet) to ducks reduced ADG and ADFI by 30
and 22% in the overall period (Adeola, 2018). Ducks fed
low nPP diet had higher F/G during day 0 to 14, but
lower F/G during day 15 to 35 without any effect on
overall F/G in the current study. Previous studies have
shown that the results of F/G in ducks fed low nPP
diet are not so consistent, especially in the grower diet.
Adeola (2018) observed increased F/G from day 1 to 15,
day 15 to 29, day 29 to 43, and the whole experiment
in ducks fed low nPP diet. Adeola (2010) demonstrated
that F/G in ducks fed low nPP diet was increased dur-
ing day 7 to 17. Zeng et al. (2015) reported higher F/G
in ducks fed low nPP diet during day 1 to 14 without
any effect on F/G during day 15 to 35. Others also ob-
served the similar negative effects of low nPP diet on
growth performance in ducks (Ravindran et al., 1995;
Orban et al., 1999; Rodehutscord et al., 2003, 2006).
It has also been well documented that low nPP diet
in broilers induced reduction in growth performance of
broilers (Cowieson et al., 2015; Taheri and Taherkhani,
2015; Farhadi et al., 2017; Pieniazek et al., 2017).
In this study, the addition of increasing residual
phytase (500 to 4,000 units/kg) in low nPP diet
improved ADG and ADFI by 50 and 47% throughout
the experiment in a dose-dependent manner. Feeding
low nPP diet (1.3 g nPP/kg) with increasing phytase
levels from 500 to 1,500 units/kg in ducks linearly
 RESIDUAL SUPERDOSES OF PHYTASE IN DUCKS 3933
Table 10. Effects of superdoses of phytase on relative organ weight in ducks fed phosphorus-deficient diets.1

Residual phytase activity, units/kg

Items4 PC2 NC2 500 1,000 2,000 3,000 4,000 SEM3 P-value

Day 14
Heart, % 6.17c 9.39a 7.68b 6.83b,c 6.96b,c 6.38c 6.12c 0.22 < 0.01
Liver, % 28.4b 36.1a 29.9b 27.6b 29.4b 27.7b 27.0b 0.64 < 0.01
Spleen, % 1.24 1.33 0.88 1.11 1.20 1.22 1.16 0.05 0.49
Lung, % 8.95 8.14 7.87 9.30 8.40 8.15 7.87 0.25 0.72
Bursa fabricius, % 2.29a 1.56b 2.73a 2.47a 2.45a 2.46a 2.51a 0.11 0.04
Day 35
Heart, % 5.82b 6.74a 5.61b 5.70b 5.19b 5.43b 5.35b 0.10 < 0.01
Liver, % 22.0b 35.1a 23.4b 22.1b 21.9b 21.2b 23.0b 0.83 < 0.01
Spleen, % 0.87 0.95 0.97 0.79 0.84 0.88 0.84 0.05 0.97
Lung, % 11.29a 7.78b 9.35a,b 9.80a,b 9.54a,b 10.59a 9.14a,b 0.35 0.04
Bursa fabricius, % 1.14 1.03 1.45 1.44 1.29 1.10 1.38 0.06 0.44
1
Means represent 5 pens of ducks with 16 birds per pen (n = 5/group).
2
PC, positive control maize-soybean meal-based diet with adequate Ca and nPP content; NC, negative control maize-soybean
meal-based diet with low nPP content.
3
Pooled standard error of the means.
4
Organ size was expressed as a percentage of BW.
a–c
Means in the same row with different superscripts differ (P < 0.05).

Table 11. Parameters of fitness between selected index and phytase, especially at high dose, have been reported
phytase retention via Broken quadratic.1 (Walk et al., 2013, 2014; Taheri and Taherkhani, 2015;
Optimal Maximum Goodness
Farhadi et al., 2017; Pieniazek et al., 2017).
Items Rate constant requirement value of fit (R2 ) The low nPP NC diet decreased performance com-
pared with the PC diet, which indicated that P was a
Day 0 to 14
ADG –7.319∗ 10−6 1,776 43.2 0.9828 limiting nutrient for the growth performance of ducks
ADFI –2.120∗ 10−5 1,323 59.9 0.9818 used in the present study. In most cases, performance
FCR –6.467∗ 10−8 1,385 0.73 0.7393 in response to increasing residual phytase superdose
Day 14 to 35
ADG –1.426∗ 10−5 1,555 109.5 0.9555 was in between that of the PC and NC groups. The
ADFI –4.539∗ 10−5 1,315 202.8 0.9308 responses of ducks in growth performance were positive
FCR –4.467∗ 10−9 1,369 0.54 0.3436 and linear or quadratic with increasing residual su-
Day 1 to 35
ADG –1.145∗ 10−5 1,599 79.8 0.9749
perdoses of phytase. Supplementation of residual 500,
ADFI –3.543∗ 10−5 1,306 139.8 0.9545 1,000, 2,000, 3,000, and 4,000 units phytase/kg diet to
FCR –4.537∗ 10−9 1,417 0.56 0.6896 NC diet increased ADG by 66, 90, 96, 110, and 102%,
Day 14 respectively, in the current study. This observation is
Tibia ash –5.339∗ 10−6 1,676 45.5 0.8863
Tibia length –9.315∗ 10−6 1,333 84.3 0.8934 also consistent with ADFI. Recently, Adeola (2018)
Tibia weight –6.660∗ 10−8 3,245 1.87 0.8236 demonstrated the similar improvement in BW gain (26
Day 35 to 47%), feed intake (15 to 26%), and BW in ducks fed
Tibia ash –3.355∗ 10−7 3,333 54.0 0.4647
Tibia length –7.842∗ 10−7 555 117.1 0.5155 diets containing 500, 1,000, 1,500, or 15,000 phytase
Tibia weight –1.214∗ 10−6 1,265 5.14 0.6932 units/kg, relative to the NC diet. Previous studies also
1
With the addition of measurement as the abscissa (x) and the
observed that BW gain and feed intake were increased
evaluation index as the ordinate (y), the quadratic broken line fitting by 5, 48, or 62% and 6, 34, or 35%, respectively, in
equation is as follows: ducks with increasing addition of 500, 1,000, and 1,500
When x < optimal dose, y = rate constant ∗ (optimal requirement- phytase units/kg in NC diets (Adeola, 2010). The con-
x)2 + maximum value.
When x > optimal dose, y = maximum value. sistent findings in broilers fed NC diets with superdoses
of phytase that the increase in the growth performance
of broilers to a level was comparable to the PC diet has
increased feed intake, feed efficiency, BW gain, and also been shown by several previous investigators (Walk
final BW (Adeola, 2010). Supplemental phytase in et al., 2014; Manobhavan et al., 2016). Interestingly,
low nPP diets (500 to 2,000 units/kg) was able to inconsistent results in F/G during day 0 to 14, day
improve growth performance in ducks (Orban et al., 15 to 35, or day 0 to 35 were observed in the present
1999; Rodehutscord et al., 2006; Adeola, 2010). In study between NC and PC groups. Others observed
agreement with our results, Adeola (2018) reported consistent increase in F/G in ducks (Adeola, 2010; Zeng
that superdoses (15,000 units/kg) of phytase increased et al., 2015; Dai et al., 2018) or broilers (Cowieson et
feed intake, feed efficiency, and BW gain in 43-day-old al., 2014, 2015; Beeson et al., 2017). Walk et al. (2014)
ducks fed diets with low nPP (3.0 g nPP/kg starter and Vieira et al. (2015) reported an increase in F/G of
diet and 2.0 g nPP/kg grower diet). In a variety of broilers fed with an inadequate-P diet. Feeding phytase
studies, similar improvements in growth performance of superdose (4,000 to 6,000 units/kg) increased F/G
broiler fed low nPP diets supplemented with increasing during day 1 to 21 in broilers (Walk et al., 2013, 2014).
3934
The use of phytase increases phytate hydrolysis
(Orban et al., 1999) and has been demonstrated to
improve nutrient utilization (Rodehutscord et al., 2006;
Adeola, 2010). Although we did not measure nutrient
digestibility in this study, the growth data suggest that
the availability of P and nutrient utilization was likely
to improve with supplemental phytase in low nPP
diets (500 to 2,000 units/kg) as the findings of previous
studies in ducks (Orban et al., 1999; Rodehutscord et
al., 2006; Adeola, 2010). Adeola (2018) reported that
superdoses (15,000 units/kg) of phytase increased ileal
digestibility and retention of P in 43-day-old ducks
fed diets with low nPP (3.0 g nPP/kg starter diet
and 2.0 g nPP/kg grower diet). Feeding low nPP diet
(1.3 g nPP/kg) with increasing phytase levels from
500 to 1,500 units/kg in ducks linearly increased the
utilization of P (Adeola, 2010). Several studies have
demonstrated that phytate-P (60 to 70% of total P in
diet) is less available to non-ruminant animals because
it can bind nutrients (minerals, Cu, starch, or AA) to
reduce utilization (Viveros et al., 2002; Pirgozliev et al.,
2012). Walk et al. (2014) observed the improvements
in performance by phytase and explained that phytase
can largely release the P in low nPP diets, eliminating
the adverse effects of phytate on digestibility of various
nutrients in broilers (Farhadi et al., 2017).
Tibia Mineralization
The tibia is usually used to evaluate mineralization
because 60 to 80% P in the skeletal tissue plays key
roles in bone mineralization (Adeola, 2018). Besides,
Zeng et al. (2015) reported that bone mineral con-
centration may be more sensitive than performance in
assessing mineral deficiency. In the current study, the
tibia length, weight, ash, Ca, and P were decreased by
low nPP diet compared with PC diet, but improved by
supplementation of NC diet with increasing residual su-
perdoses of phytase, which was in agreement with pre-
vious studies. Feeding low nPP diet decreased tibia ash
and mineralization in ducks, whereas bone mineral con-
tent, bone density, or bone ash increased linearly when
diets were supplemented with phytase (Rodehutscord
and Dieckmann, 2005; Adeola, 2010, 2018; Zeng et al.,
2015). It is speculated that low P intake due to low
nPP diet in ducks reduced the synthesis and deposi-
tion of hydroxyapatite and bone mineralization in the
tibia. Increases in tibia ash percentage were good in-
dication of increased bone mineralization. There were
linear and quadratic effects of residual phytase super-
dose supplementation to the low nPP diet on tibia P.
The low nPP diet decreased tibia ash much less on
day 35 (6%) compared with that on day 14 (32%) in
this study, which indicated that age might also be a
factor. The present study found a consistent improve-
ment in growth performance and tibia mineralization
with the increasing residual phytase superdose (500 to
4,000 units phytase/kg) compared with NC diet, which
FAN ET AL.
implied increased P digestibility and utilization. More-
over, addition of phytase in low nPP diets increased
the retention of P, K, Na, Mg, S, Cu, Fe, and Mn
(Cowieson et al., 2006) and the availability of P, Ca,
Zn, and Cu (Sebastian et al., 1996) in broilers. This is
supported by previous studies that demonstrated the
improvements may be attributed to hydrolysis of phy-
tate and the removal of antinutritive effects (Shirley and
Edwards, 2003), higher nutrient digestion and retention
(Dos Santos et al., 2013; Walk et al., 2013, 2014), in-
creased immunity and antioxidant status (Liu et al.,
2008, 2010a; Karadas et al., 2010), altered gene expres-
sion associated with growth (Liu et al., 2010b; Woyengo
et al., 2011), and decreased excreta moisture content
(Delezie et al., 2015). Most of the tibia minerals of ducks
fed diets with residual phytase superdose were nearly
equivalent to those fed PC diet in the current study.
Similar to the results of the present study, numerous
studies about nPP diet in broilers found tibia strength
and ash content were reduced by low nPP diet, while
increased by phytase addition (Cowieson et al., 2014,
2015; Akter et al., 2016). Nevertheless, the reason for
the increase in tibia Fe for NC diet increased on day 14
and 35 was not clear.
Relative Organ Weight
Measurement of digestive and immune organ weights
may reflect the digestive and immune status in ducks
(Feng et al., 2017). In the current study, birds fed NC
diets had greater duodenum, jejunum, ileum, and ce-
cum index on day 14 and 35, which indicated that ducks
were sensitive to low nPP diet among all the digestive
organs. This may mirror the changes in F/G. The in-
crease in intestinal index may also be due to decreased
BW. Supplementation of increasing residual phytase su-
perdose counteracted the negative effects of low nPP
diet on intestine development with the increasing BW.
The relative weights of heart, liver, lung, spleen, and
bursa fabricius in ducks fed NC diets with increasing
residual phyatse superdoese were comparable to those
in ducks fed PC diet. To our knowledge, this is the first
study to determine the effects of low nPP diet and the
residual phyatse superdose on relative organ weight. It
was supposed that the negative effects of low nPP on
intestine may be related to its influence on bacterial
community in the gut (Dai et al., 2018). It is suggested
that ducks fed low nPP diet had poor growth perfor-
mance, but higher α-diversity of cecal microbial popu-
lation because they need spend more energy to compete
with the gut microbial population to obtain more P and
other nutrients (Dai et al., 2018). The regulation of the
dietary P supply could improve growth performance
of ducks by regulating gut health (Dai et al., 2018),
which may reflect the improvement in growth perfor-
mance in the present study. However, studies about the
interaction between nPP levels and gut microbiota of
ducks are poorly understood. Therefore, more studies
 RESIDUAL SUPERDOSES OF PHYTASE IN DUCKS
are needed to demonstrate the hypothesis that low nPP
diet could affect gut bacterial colonization and impair
gut health of ducks and addition of residual phytase
superdose could counteract these negative effects.
Optimal Requirement
Based on growth performance and tibia quality as
a response of supplemental residual phytase superdose,
the optimal requirement of residual phytase superdose
ranged from 555 to 3,333 units/kg diet for all traits
with the best supplementation levels around 1,306 to
1,777 units/kg diet for growth performance. In agree-
ment with the results of performance, ducks fed NC
diets with residual 3,000 units phytase/kg diet during
day 0 to 14, 1,000 units/kg during day 15 to 35, and
2,000 units/kg during the overall period could achieve
the performance comparable to those fed PC diet.
CONCLUSIONS
Feeding low nPP diet (2.7 g nPP/kg diet reduc-
tion compared with than PC) led to adverse effects
on growth performance, tibia mineralization, and rel-
ative organ weight compared with the PC diet, which
indicated that P was indeed a limiting nutrient in the
diet. Supplementation of residual superdoses of phytase
consistently improved ADG, ADFI, and tibia mineral-
ization during the whole experiment. Moreover, resid-
ual phytase superdose at 4,000 units/kg did not cause
detrimental effects on growth performance, tibia min-
eralization, or relative organ weight in ducks. However,
the data suggest that growth performance of ducks fed
NC diets with residual 3,000 units phytase/kg diet dur-
ing day 0 to 14, 1,000 units/kg during day 15 to 35, and
2,000 units/kg during the overall period was compara-
ble to PC diet in this study. Correspondingly, the opti-
mal requirement of residual phytase superdose ranged
from 1,306 to 1,777 units/kg diet for growth perfor-
mance based on the quadratic broken line model.
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