Influence of particle size and calcium source on production performance,

egg quality, and bone parameters in laying ducks

S. Wang, W. Chen, H. X. Zhang, D. Ruan, and Y. C. Lin1

Institute of Animal Science, Guangdong Academy of Agricultural Sciences, State Key Laboratory of Livestock
and Poultry Breeding, Key Laboratory of Animal Nutrition and Feed Science in South China,
Ministry of Agriculture Guangdong Public Laboratory of Animal Breeding and Nutrition Guangdong,
Key Laboratory of Animal Breeding and Nutrition Guangzhou 510640, China

ABSTRACT The influence of calcium source (limestone
and oyster shell) and particle size (<0.1 mm; 0.85 to
2 mm) on laying performance, egg quality, and bone
properties were examined in laying ducks by a 2 × 2
factorial arrangement of treatments. Longyan females
(288) with similar BW at 24 wk of age were randomly
allotted into 4 treatments, each with 6 replicates of 12
individually caged birds and studied over the follow-
ing 12 wk. Particle size affected egg weight and feed
conversion (P < 0.05). Large particle size increased
shell breaking strength, albumen height, Haugh unit,
and shell content of phosphorus and magnesium (P <
Key words: calcium source, particle size, productive performance, egg quality
0.05), but had no effect on egg shape, yolk color, shell
thickness, or the weight proportion of shell. There were
no effects of particle size on tibial properties: dry de-
fatted weight, calcium content, or breaking strength.
Limestone increased albumen height, shell content of
calcium and phosphorus, and the breaking strength of
tibia (P < 0.05). It is concluded that limestone with
a large particle size provided for superior productive
performance, egg quality, and bone characteristics and
is more suitable than oyster shell for practical applica-
tions.
2014 Poultry Science 93:2560–2566
http://dx.doi.org/10.3382/ps.2014-03962

INTRODUCTION be used as a complete calcium source for laying hens

Calcium is a critical factor for egg production, but
little is known about the optimal way of providing this
for laying ducks. Many studies with hens have shown
that calcium sources with large particle size prolong
the residence time of the particles in the stomach of
poultry, favoring solubilization so that calcium ions are
made available in sufficient quantities for absorption.
Zhang and Coon (1997a) noted that large particle size
limestone (>0.8 mm) with lower solubility in vitro (30
to 50%) was retained in the gizzard for a longer time,
thereby increasing its solubility in vivo to as much as
94%. Rao and Roland (1989) showed that an increased
proportion of calcium was solubilized from larger par-
ticles of limestone in the digestive system during a 25-h
period, providing greater availability of calcium ions
for absorption. Additional sources of calcium have been
introduced and are in practical use with birds, such
as egg shell, oyster shell, and others. Gongruttananun
(2011) indicated that ground, sterilized eggshell could
©2014 Poultry Science Association Inc.
Received February 10, 2014.
Accepted June 25, 2014.
1 Corresponding author: lyc0123@tom.com
and Safaa et al. (2008) showed that 40% oyster shell,
substituted for limestone, did not affect the laying per-
formance of brown-shell laying hens. There are few
studies on the use of 100% oyster shell as the calcium
source and none examining different of calcium sources
and particle sizes for laying ducks. A large number of
experiments has been carried out in hens, but the lit-
erature for ducks is very limited. The crop of ducks is
just a distensible gullet, whereas that of hens is a stor-
age organ. The ovipository cycle and laying time also
differ between ducks and hens. These differences may
prevent simple extrapolation of findings on calcium
sources and different particle size from hens to ducks.
The present study has investigated the effect on laying
performance, egg quality, and tibial indices of oyster
shell or limestone, each of 2 particle sizes as calcium
sources for laying ducks.
MATERIALS AND METHODS
Experimental Design, Birds, and Housing
This study was approved by the Animal Care and
Use Committee of Guangdong Academy of Agricul-
ture Science. Longyan ducks (288), of the same genetic

2560
 PARTICLE SIZE AND CALCIUM SOURCE FOR DUCKS 2561
Table 1. Composition and nutrient levels of the diets (g/kg, as fed basis)
Small particles (SP) Large particles (LP)

Item Limestone Oyster shell Limestone Oyster shell

Ingredient
 Corn 552.9 553.1 554 555
  Soybean meal 242 244 245 244.5
  Wheat bran 87 82.7 79 80.3
  Calcium hydrogen phosphate 12.2 12.2 12.3 12.2
  Limestone (LP 34.23%)1 — — 94 —
  Oyster shell (LP 34.85%)1 — — — 92.3
  Limestone (SP 35.65%)1 90.2 — — —
  Oyster shell (SP 34.85%)1 — 92.3 — —
  dl-Methionine 1.65 1.65 1.65 1.65
  l-Lysine·HCl 0.55 0.55 0.55 0.55
  Sodium chloride 3.5 3.5 3.5 3.5
 Premix2 10 10 10 10
 Total 1,000 1,000 1,000 1,000
Calculated analysis3        
  AME (kcal/kg) 2.50 2.50 2.50 2.50
  CP (%) 17.25 17.83 17.46 17.17
  Total Lys (%) 0.87 0.87 0.87 0.87
  Total Met (%) 0.43 0.43 0.43 0.43
  Ca (%) 3.45 3.37 3.49 3.28
  Total P (%) 0.57 0.54 0.54 0.52
  Available P (%) 0.35 0.35 0.35 0.35
1Measured values of calcium content.
2Provided the following (per kg of diet): vitamin A, 12,000 IU; vitamin D3, 2,000 IU; vitamin E, 38.7 IU; vitamin K3, 1.0 mg; vitamin B1, 3.0 mg;
vitamin B2, 9.6 mg; vitamin B6, 6.0 mg; vitamin B12, 0.03 mg; choline chloride, 1,000 mg; d-pantothenic acid, 28.5 mg; folic acid, 0.6 mg; biotin, 0.15
mg; Fe, 50 mg; Cu, 10 mg; Mn, 90 mg; Zn, 90 mg; I, 0.50 mg; Se, 0.40 mg.
3The CP, Ca, and total P are measured values in the mixed feeds.

background and of comparable BW at 24 wk of age,
were assigned at random to 4 experimental groups,
each containing 6 replicates of 12 individually caged
(27.8 × 40 × 55 cm3) birds and studied for 12 wk.
Each replicate consisted of 2 banks of 6 adjacent cages
and each bank shared a deep feed trough enabling in-
dividual feeding of the 6 birds without spillage at 0700
and 1500 h twice daily. Birds were fed the same, thor-
oughly mixed corn-soybean-based diet, formulated for
laying ducks (Table 1), but with 4 different calcium
components (see below), each providing 3.6% calcium
and exceeding the NRC recommendation (NRC, 1994).
The daily allowance of feed (average 144 g) was the
maximum for individuals without their leaving refus-
als. A 2 × 2 factorial arrangement was used to compare
the calcium sources, ground limestone or oyster shell,
each provided at 2 particle sizes. Each calcium source
was purchased commercially and ground coarsely and
finely. Large particles (LP) were between 1.18 and 2
mm (>84%), as determined by sieve analysis using a vi-
bratory shaker and small particles (SP) were almost all
less than 0.1 mm (Table 2). The ducks were all housed
in the same room, with incandescent lighting of 10 lx,
providing 15L:9D. After 12 wk of feeding, 2 birds of
average BW for the treatment group were selected from
each replicate and heparinized blood was collected from
the wing vein at 1000 h, 7 to 9 h after the previous ovi-
positioning, when the next ovum is in the shell gland.
Those birds were then stunned and exsanguinated, and
the tibias were collected.
Productive Performance
The birds were fed the 4 diets for a 7-d adaptation
period after which feed intake was recorded daily on a
per replicate basis (2 troughs). Likewise, the number of
total, broken, and shell-less eggs were recorded daily by
replicate. All eggs produced were individually weighed
and graded daily (European Economic Community,
1989). Egg production, egg weight, egg yield (average
egg weight per day per duck), ADFI, and feed conver-
sion ratio (FCR, feed/eggs) were calculated daily from

Table 2. Particle size distribution of limestone and oyster shell

Small particles (%) Large particles (%)

Size (mm) Limestone Oyster shell Size (mm) Limestone Oyster shell

>0.18 0.36 0.82 >2 1.1 0.38

0.15 to 0.18 0.2 0.34 1.18 to 2 95.32 84.96
0.1 to 0.15 1.06 1.02 0.85 to 1.18 3.56 13.52
<0.1 98.28 97.7 <0.85 0.11 1.14
2562 Wang et al.
these data but are presented as the averages for the
complete 12-wk period.
Egg Quality
Egg quality was measured on 8 eggs collected at
random from each replicate once weekly during wk 9
to 12, and the average of 32 eggs from each replicate
was calculated. Yolk color, albumen height, and Haugh
units were measured on the day of collection using an
Egg Analyzer (model EA-01, ORKA Food Technology,
Ramat HaSharon, Israel). The strength of the shell
was determined on the vertical axis using an Egg Force
Reader (model EFR-01, ORKA Food Technology). Egg
length, egg width, and eggshell thickness were mea-
sured using a digital micrometer, the distance between
the blunt and pointed ends for length, and the diameter
at mid-length for the width. The egg shape index was
calculated as egg length divided by width. The shells
with membranes were weighed, washed under a gentle
flow of water to remove adhering albumen, then ashed
at 700°C. Shell (1 g) was heated with 15 mL of nitric
acid and 2 mL of perchloric acid until the appearance
of white fumes then quantitatively diluted to 100 mL
with water. Calcium was determined by titration with
EDTA and phosphorus by ammonium molybdenate and
UV spectrophotometry. Magnesium was determined by
atomic absorption spectrometry (Hitachi/Z-2000, Hita-
chi Limited, Tokyo, Japan), as described by Alfrey et
al. (1974).
Plasma Analysis
Freshly collected blood (2 samples from each repli-
cate) was centrifuged (1,200 × g) at 4°C for 10 min and
plasma was held at –20°C until used. Plasma calcium,
phosphorus, and alkaline phosphatase contents were
determined for all samples by an automatic biochemi-
cal analyzer (Synchron cx5, Beckman Coulter Inc., Los
Angeles, CA), with reagents from BioSino Bio-Technol-
ogy and Plasma Science Inc. (Beijing, China).
Tibial Characteristics
Two pairs of tibias were collected from each replicate
for analyses. The left tibias were ashed for 24 h, and the
content of tibial ash was determined on a dry defatted
weight basis (Lin and Shen, 1979). Calcium and phos-
phate in bone ash were determined, as described above
for shell. Right tibias were cleaned of all tissue and
bone mineral density of the right tibia was measured
at the Guangzhou Overseas Chinese Hospital with a x-
ray osteodensitometer (Lunar Prodigy, General Electric
Company, Fairfield, CT) and the breaking strength of
the middle of the bone was determined with a materials
tester (Instron 4411, Instron Corporation, Grove City,
PA) using software version 8.09, a standard 50-kg load
cell, and a modified shear plate (8 cm in length and 1
mm in width), as described by Riczu et al. (2004).
Scanning Electron Microscopy
The morphology of freshly fractured, nondecalcified
eggshell fragments from the limestone diets with SP
and LP was evaluated. For each treatment, 12 shell
fragments from the blunt end were air-dried, mounted
with conductive carbon cement, scanned, and imaged
with an environmental scanning electron microscope
(XL-30-ESEM, FEI, Eindhoven, the Netherlands) at
an accelerating voltage of 20 kV; stubs were oriented
to provide a transverse view of the eggshell (Figure 1).
Images of transverse sections of eggshell from ducks fed
limestone with LP and SP are shown at 400×. The crys-
tal layers are shown at higher magnifications (6,400×).
Statistical Analysis
Replicate (n = 6) was taken as the experimental unit.
Except where noted otherwise, 2 sampled birds per rep-
licate were used. The effects of source and particle size
were analyzed by the 2-way ANOVA procedure of SAS
(SAS Institute Inc., 2003) with P < 0.05 indicating sig-
nificance. Analyses used the following model:
Yijk = μijk + Si + PSj + (S × PS)ij + εijk,
where Yijk is the replicate value of a given variable, μijk
is the mean, Si is the effect of calcium source (i = 1 or
2, limestone or oyster shell), PSj is the effect of particle
size (j = 1 or 2, LP or SP), (S × PS)ij is the effect of in-
teraction between the 2 main effects, and εijk is the ran-
dom error. No significant interactions were detected, so
the model was reduced to just the 2 main effects. For
some variables, pooling or averaging within replicates
over time was used as there was no experimental inter-
est in possible temporal changes.
RESULTS
Productive Performance
There was no statistical difference between the slight-
ly higher egg production of ducks on the LP than the
SP diets (Table 3). Increasing the particle size from SP
to LP increased egg yield and decreased FCR (both P
< 0.05). The source of calcium had no effect on these
indices of productive performance over the whole 12-wk
period. There was no mortality during the experiment.
Egg Quality
As shown in Table 4, LP increased the breaking
strength of the shell (P < 0.05), and improved the ap-
pearance of eggs based on increased albumen height
and Haugh units (P < 0.01). Particle size did not af-
fect egg shape index, yolk color, eggshell thickness, or
weight. The LP diets resulted in higher phosphorus and
lower magnesium contents of eggshells (P < 0.01) than
did SP diets, but calcium content was unaffected. The
 PARTICLE SIZE AND CALCIUM SOURCE FOR DUCKS 2563
Table 3. Effect of calcium source and particle size on productive performance in laying ducks
Size1 Source P-value3

Item SP LP Limestone Oyster shell SEM2 Size Source

Egg production (%) 93.39 95.72 94.89 94.22 1.41 0.11 0.64
Egg yield (g/d) 60.78 62.35 61.87 61.35 0.71 <0.05 0.48
Feed conversion4 (g/g) 2.39 2.33 2.35 2.37 0.02 <0.05 0.34
1Smaller particles (SP) and larger particles (LP).
2Derived from ANOVA error mean square for n = 6 replicates.
3No interactions were significant.
4All ducks were given 144 g/d of feed.

source of calcium affected the albumen height, shell cal-
cium content, and shell phosphorus content (P < 0.05),
both being higher with the limestone diet. There were
no differences between limestone and oyster shell on the
remaining indices of egg quality.
Plasma Variables and Tibial Characteristics
Particle size and sources of Ca did not significantly
affect plasma calcium, phosphorus, or alkaline phos-
phatase activity (Table 5). The larger particle size of
the calcium sources increased (P < 0.05) the breaking
strength of fresh tibia, dry weight, and calcium content
of defatted tibia of the ducks (Table 6). The slightly
greater density of the defatted tibia and increased tibial
ash and phosphorus in the diets with LP were not sig-
nificant. Inclusion of limestone rather than oyster shell
did increase the breaking strength of the tibias (P <
0.05), but not the other tibial characteristics. Overall,
the best tibial indices were obtained from use of lime-
stone of large particle size.
Scanning Electron Microscopy
Typical images of scanned transverse sections of
eggshells from ducks fed the limestone diets, LP and

Figure 1. Environmental scanning electron microscope images of transverse sections of eggshell from ducks fed limestone with larger particles
(LP) is shown at 400× in panel a, and from ducks fed limestone with smaller particles (SP) in panel b. Higher magnifications, showing the crystal
layer (6,400×), indicate greater regularity with the LP diet (panel c) than the ragged, loose organization from the SP diet (panel d). Arrows in
panel a indicate the shell membrane (SM) and mammillary layer (ML).
2564 Wang et al.
Table 4. Effect of particle size and source of calcium on egg quality in laying ducks
Size1 Source P-value3

Variable SP LP Limestone Oyster shell SEM2 Size Source

Shape index 1.37 1.36 1.36 1.37 0.01 0.60 0.52

Breaking strength (kg) 3.87 4.13 3.99 4.01 0.12 <0.05 0.87
Albumen height (mm) 5.38 6.02 5.88 5.52 0.17 <0.01 <0.05
Yolk color 4.11 4.17 4.17 4.11 0.10 0.60 0.60
Haugh units 70.67 75.93 74.75 71.85 1.44 <0.01 0.06
Shell thickness (mm) 0.33 0.32 0.32 0.33 0.03 0.15 0.24
Shell weight (g) 6.17 6.18 6.16 6.20 0.49 0.87 0.72
Shell calcium (% DM) 36.12 36.34 36.47 35.99 0.20 0.29 <0.05
Shell phosphorus (% DM) 0.146 0.195 0.185 0.156 0.01 <0.01 <0.01
Shell magnesium (% DM) 0.160 0.148 0.155 0.153 0.003 <0.01 0.623
1Smaller particles (SP) and larger particles (LP).
2Derived from ANOVA error mean square for n = 6 replicates.
3No interactions were significant.

SP, are shown in Figure 1. The overall structure of the
eggshell from the LP diet (Figure 1a) appeared to be
dense and pyknotic and showed close arrangement,
tightly connected into a mass with fuzzy boundaries.
Eggshell from the SP diet was less substantial with
a loose arrangement of clearly discernible mammillae
(Figure 1b). Connection was poorer, with obvious gaps,
between the mammillary layer and shell membranes.
The crystal layer of the LP eggshell was more regular
(Figure 1c), whereas that of the SP was ragged and
loose (Figure 1d).
DISCUSSION
Effect of Particle Size on Productive
Performance, Egg Quality, Tibial
Characteristics, and Scanning Electron
Microscopy of Eggshell
The present study demonstrated a positive effect on
most productive traits, including egg yield and FCR,
in laying ducks with intake of larger particle size of
the calcium source. There is little information in sup-
port of this observation in the available literature, and
nothing for ducks. Skřivan et al. (2010) showed that
feeding younger and older hens a calcium source with
large particle size (0.8 to 2 mm) increased egg produc-
tion, egg weight, and FCR. Koreleski and Swiatkiewicz
(2004) observed a nonsignificant increase in hen-day
egg production (from 90.1 to 91.8%) when one-half of
the fine limestone (<0.4 mm) was replaced with par-
ticulate limestone grit (2.0 to 4.0 mm). Consistent with
Koreleski and Swiatkiewicz (2004), who found that the
limestone grit increased eggshell breaking strength, the
present results using ducks demonstrate greater break-
ing strength and enhanced egg appearance. A similar
effect on albumen height and Haugh unit was shown
in chickens fed limestone of large particle size (Skřivan
et al., 2010). Little is known about factors influencing
the mineral composition of the shell so the reasons why
shell %P was increased by LP and by limestone, whereas
%Mg decreased with LP are not obvious. There are re-
ports (Guinotte and Nys, 1991; Pizzolante et al., 2009;
Araujo et al., 2011) indicating no significant effect of
limestone particle size on productive performance and
egg quality.
In the ducks studied here, particle size of the cal-
cium source influenced the quality of bone, indicated
by breaking strength, dry defatted weight, and calcium
content of the tibia, but not that of calcium, in egg-
shells. This outcome was consistent with particulate
limestone increasing accumulation of medullary bone
in the proximal tarsometatarsus, improved breaking
strength of the tibia and radiographic densities of tibia
and keel (Fleming et al., 1998). Similarly, Saunders-
Blades et al. (2009) showed that a mixed particle size
calcium source (1/3 particulate) improved tibial weight,
density, mineral content, and breaking strength. Some-
what different results have also been obtained with

Table 5. Effect of particle size and source of calcium on plasma variables in laying ducks
Size2 Source P-value4

Variable1 SP LP Limestone Oyster shell SEM3 Size Source

Ca (mM) 3.50 3.43 3.35 3.57 0.16 0.67 0.19

P (mM) 2.28 1.91 1.85 2.34 0.32 0.26 0.16
AKP (U/L) 111 97 112 96 1.59 0.26 0.16
1Alkaline phosphatase (AKP).
2Smaller particles (SP) and larger particles (LP).
3Derived from ANOVA error mean square for n = 6 replicates.
4No interactions were significant.
 PARTICLE SIZE AND CALCIUM SOURCE FOR DUCKS
Table 6. Effect of calcium source and particle size on tibial characteristics in laying ducks
Size1
Variable SP LP
Breaking strength (N) 166.4 178.8
Density (g/cm2) 0.26 0.30
Dry defatted weight (g) 2.97 3.24
Tibia ash (%) 60.64 61.95
Tibia P (%) 10.32 10.58
Tibia Ca (%) 21.97 22.86
1Smaller particles (SP) and larger particles (LP).
2Derived from ANOVA error mean square for n = 6 replicates.
3No interactions were significant.
hens, such as increased shell calcium (as well as phos-
phorus) from large particle limestone (Skřivan et al.,
2010) and a conditional effect of particle size on tibial
strength (Koreleski and Swiatkiewicz (2004).
Larger particle size limestone (>0.8 mm) with low-
er in vitro solubility (30 to 50%) was retained in the
gizzard for a longer time, which increased the in vivo
solubility (Rao and Roland, 1989; Zhang and Coon,
1997a). The longer retention time increases the dura-
tion of providing Ca to the intestine, allowing a more
consistent supply of calcium during the night when the
calcification takes place. Although this could account
for the beneficial effects of LP found here in ducks,
it did not result in any change in plasma calcium or
phosphorus measured at 1000 h, but eucalcemia is
tightly regulated. Net enteric uptake of calcium was
not measured here, but the presumed greater availabil-
ity from the LP sources could account for the increase
observed in tibial calcium. Bain (1992) found that the
fracture toughness of the eggshell was a measure of its
resistance to unstable crack growth, and in this respect
there appeared to be a link between the latter and the
structural organization of the mammillary layer. Larger
particle size clearly improved the mammillary layer of
the duck’s eggs, which could explain the significant in-
crease in their mechanical strength found here.
Effect of Calcium Source on Productive
Performance, Egg Quality, and Tibial
Characteristics
The source of calcium, limestone, or oyster shell did
not generally affect productive performance, egg qual-
ity, or tibial characteristics with the exception of albu-
men height, shell content of calcium and phosphorus,
and breaking strength of the tibia, all of which were
higher with limestone. Most published research with
hens has the same findings. For example, Richter et al.
(1999) found that various calcium sources (limestone,
limestone together with oyster shells, marine shells, and
shells treated with phosphoric acid) did not affect feed
intake, laying performance, egg weight, feed conversion,
or mortality. There were minimal influences on stabil-
2565
Source P-value3
Limestone Oyster shell SEM2 Size Source
180.3 164.9 5.81 <0.05 <0.05
0.28 0.28 0.02 0.10 0.86
3.16 3.05 0.12 <0.05 0.33
61.23 61.36 0.84 0.13 0.88
10.44 10.45 0.15 0.10 0.92
22.36 22.48 0.35 <0.05 0.74
ity variables of eggshells such as deformation, ultimate
strength, weight/100 g of eggs, shell weight/cm2, cracks,
and eggs without shells. Similarly, Safaa et al. (2008)
showed that 40% substitution of limestone with oyster
shell did not affect productive performance or egg qual-
ity of brown-shell laying hens. In contrast to the present
findings, Çatlı et al. (2012) indicated that partial sub-
stitution of limestone with oyster shell meal in laying
hens resulted in significant increases in egg production
(numbers and mass) and greater feed intake. Earlier
studies concerning oyster shell meal supplementation
showed beneficial effects on eggshell breaking strength
(Brister et al., 1981; Makled and Charles, 1987; Proud-
foot and Hulan, 1987; Guinotte and Nys, 1991; Grizzle
et al., 1992; Ahmad and Balander, 2004).
In the ducks studied here, oyster shell decreased albu-
men height and marginally (P = 0.06) reduced Haugh
unit, suggesting that it compromised freshness of the
eggs. Because the oyster shells sometimes contained
meat residues, there is the possibility of microbial con-
tamination in shells and most research indicates that
oyster shell is less soluble than limestone (Zhang and
Coon, 1997b; Lichovnikova, 2007). Limestone and oys-
ter shell of the same particle size would be expected
to experience similar retention in the digestive tract,
so digestibility of calcium and phosphorus must differ.
It is assumed that reduced net absorption of calcium
and phosphorus in ducks fed oyster shell resulted in
decreased shell contents of these elements and break-
ing strength of the tibia compared with ducks given
limestone.
In conclusion, limestone with large particle size was
shown to increase productive performance, and improve
the quality of eggs and bone. Calcium sources with large
particle size, rather than finely ground, significantly
improved productive performance, egg appearance, egg
quality, and the quality of the tibia in ducks in heavy
production. There is no specific information regarding
particle size and calcium source in NRC (1994). Oyster
shell could be used as a substitute for limestone with a
small decrease in egg quality and tibial characteristics.
On the basis of all variables considered here, limestone
of larger particle size was the better calcium source for
laying ducks.
2566 Wang et al.
ACKNOWLEDGMENTS
The authors sincerely thank W. Bruce Currie (emeri-
tus professor, Cornell University, Ithaca, NY) for his
help in presentation of this manuscript. This work was
supported by the Earmarked Fund for Modern Agro-
industry Technology Research System (CARS-43-13),
National Natural Science Foundation of China (Grant
No. 31301995), and Technology Research of Animal
Feed of the Guangdong Academy of Agricultural Sci-
ences (2011A020102009).
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