Accepted Manuscript

Title: Effect of different levels of palygorskite inclusion on

pellet quality, growth performance and nutrient utilization in
broilers
Author: Lei Zhang Rui Yan Ruiqiang Zhang Chao Wen
Yanmin Zhou
PII:
DOI:
Reference:

S0377-8401(16)30333-9
http://dx.doi.org/doi:10.1016/j.anifeedsci.2016.10.019
ANIFEE 13657

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Animal

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7-7-2016
21-10-2016
29-10-2016

Feed

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Technology

Please cite this article as: Zhang, Lei, Yan, Rui, Zhang, Ruiqiang, Wen, Chao, Zhou,
Yanmin, Effect of different levels of palygorskite inclusion on pellet quality, growth
performance and nutrient utilization in broilers.Animal Feed Science and Technology
http://dx.doi.org/10.1016/j.anifeedsci.2016.10.019
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Effect of different levels of palygorskite inclusion on pellet quality, growth

performance and nutrient utilization in broilers

Lei Zhang, Rui Yan, Ruiqiang Zhang, Chao Wen, Yanmin Zhou*

College of Animal Science and Technology, Nanjing Agricultural University,

Nanjing 210095, P. R. China

* Corresponding author. Tel.: +86 025 84396067; fax: +86 025 84395314
E-mail address: zhouym6308@163.com (Y.M. Zhou)

Highlights

1. The quality of pellets was improved with palygorskite inclusion.

2. The palygorskite supplementation improved growth performance of broilers.
3. The optimal level of palygorskite inclusion in the pellet diets was 10 g/kg.

ABSTRACT
The aim of the present study was to investigate the effects of different levels of
palygorskite on the pellet quality, growth performance and nutrient utilization in
broilers. Three hundred one-day-old Arbor Acres broilers were randomly divided into
five treatment groups with six replicates of ten broilers each for a 42-day feeding trial.
The birds were fed diets supplemented with 0, 5, 10, 15 and 20 g/kg palygorskite,
respectively. The results showed that the hardness and durability index of pellets were
improved (linear and quadratic, P < 0.001) with the increasing level of palygorskite
inclusion. The palygorskite supplementation improved average daily gain (ADG) and
average daily feed intake (ADFI) of broilers (linear and quadratic, P < 0.001). The
broilers fed the diet supplemented with 10 g/kg palygorskite had the highest ADG
among groups. Compared with the control group, neither 5 g/kg nor 10 g/kg
palygorskite inclusion affected feed per unit gain (F/G) of broilers. However, the F/G
in grower period increased 1.6% with diet 15 g/kg (1.83 vs 1.80) and 3.3% with diet
20g/kg (1.86 vs 1.80). The results indicated that palygorskite inclusion could enhance
the pellet hardness and durability index, and improve the growth performance of
broilers. In additional, the optimal level of supplemented palygorskite in the pellet
diets of broilers was 10 g/kg.
Abbreviations: ADFI, average daily feed intake; ADG, average daily gain; F/G, feed
per unit gain; PDI, pellet durability index; DM, dry matter; OM, organic matter; CP,
2

crude protein; EE, ether extract; AIA , acid-insoluble ash.

Keywords: Palygorskite; Pellet quality; Growth performance; Nutrient utilization;

Broilers
1. Introduction
The physical form of feed (mash, pellet and crumble) is a key factor in meat
yield of broiler (Rezaian et al., 2007; Ahmed and Abbas, 2013; Sena et al., 2014). It is
generally accepted that the feeding of pellets improves broiler growth performance
compared to mash (Choi et al., 1986; Sena et al., 2014). Pelleted feed also has the
benefits of decreased ingredient segregation, increased digestibility, destruction of
pathogenic organisms, thermal modification of starch and protein and increased
palatability (Abdollahi et al., 2013), but pellet feeding alone is not enough to ensure
enhanced performance of poultry. The quality of pellets must be taken into account
also (Corzo et al., 2011; Lilly et al., 2011). In turn, pellets often disintegrate,
especially when high proportions of fat are included (Angulo and Brufau, 1995;
Catal-Gregori et al., 2009). By virtue of excellent adsorption and rheological
properties, clays were used as a pellet binder to improve pellet durability index and
hardness (Angulo et al., 1996; Pappas et al., 2010; Zhang et al., 2015).
Palygorskite is a naturally available nanorod-like silicate clay mineral composed
of

ribbons

of

2:1

phyllosilicate

units

with

an

theoretical

formula

of

Si8Mg8O20(OH)2(H2O)44H2O (Bradley, 1940; Galn, 1996). Each ribbon is connected

to the next by the inversion of SiO4 tetrahedron along a set of Si-O-Si bonds, forming
zeolite-like channels with a size of about 0.37 nm0.64 nm (Chisholm, 1990;
Chisholm, 1992; McKeown et al., 2002). The special pores and channels, fine particle
size and fibrous habit of palygorskite endow it with high surface area, ion-exchange

and adsorption ability, rheological and catalytic properties (Galn, 1996; Murray,
2000). Palygorskite is applicable for use in many industrial areas. As an additive,
palygorskite is effective in binding noxious compounds and then expelling them from
the body of animals (Phillips, 1999), treating gastroenteric diseases and creating an
intestinal barrier in both animals and humans (Szajewska et al., 2006). The recent
studies have found that dietary supplementation of palygorskite could improve growth
performance, intestinal integrity, nutrient utilization and litter quality in animals
(Pappas et al., 2010; Zhang et al., 2013; Tang et al., 2014; Lv et al., 2015).
Shurson et al. (1984) found silicate minerals, as a nonnutritive additive, tend to
reduce the digestible energy and metabolizable energy. Lv et al. (2015) reported that a
lower dietary concentration of palygorskite (2000 mg/kg) was more effective than a
greater concentration (3000 mg/kg) to improve growth performance and increased dry
matter, energy and crude protein utilization. These data seem to indicate that a
relatively low concentration of dietary clay mineral may be more effective than a high
dose. Traditionally, clays have been incorporated in diets (10-20 g/kg) as a
technological additive to improve feed manufacture (Angulo and Brufau, 1995). The
addition of clays at lower level was not sufficient to optimize pellet quality, especially
when high proportions of fat are included (Angulo et al., 1996). Therefore, the aim of
this study was to investigate the effects of different amounts of palygorskite on feed
technological characteristics, growth performance, and nutrient utilization of broilers.
2. Materials and methods
2.1. Materials
Natural palygorskite was provided by Jiangsu Sinitic Biotech. Co., Ltd. (Xuyi,
Jiangsu, China) as a powder (AFG-200 mesh). The chemical composition was
determined using an Epsilon 5 X-ray Fluorescence (XRF) spectrophotometer

(PANalytical, Netherland). The major chemical components of palygorskite are

showed in Table 1.
2.2. Birds and diets
Three hundred one-day-old hateched Arbor Acres broilers with initial body
weight of 40.2 0.3 g were obtained from a commercial hatchery (Hewei Group Co.,
Ltd, Anhui, China) and randomly divided into five treatment groups with six
replicates of ten broilers for a 42-day feeding trial. The starter and grower diets (Table
2) were offered to the chickens from 1 to 21 days and 22 to 42 days, respectively. The
palygorskite levels were 0, 5, 10, 15 and 20 g/kg. Corn was ground in a hammer mill
(SFSP, Liyang Yuda Machinery Co., Ltd., Jiangsu, China) to pass through screen sizes
of 2.0 mm. The rest of ingredients (soybean meal, wheat middling, corn gluten meal,
premix, amino acids, minerals, salt and palygorskite) as mash were added directly into
the mixer. The added soybean oil was sprayed into the moving mass of dry ingredients
in the mixer via two jets. Diets were mixed in a double-screw paddle mixer (SSHJ,
Liyang Yuda Machinery Co., Ltd., Jiangsu, China). Following mixing, the mash was
steam-conditioned to 70C for 30 s and pelleted using a pellet mill (SZLH508, Liyang
Yuda Machinery Co., Ltd., Jiangsu, China). Dimensions of the die ring were 2.5 mm
diameter and 25 mm thickness and 4.0 mm diameter and 40 mm thickness for starter
and grower diets, respectively.
2.3. Experimental procedure and sampling
All procedures about broiler were approved by the Nanjing Agricultural
University Institutional Animal Care and Use Committee. All broilers were placed in
three-layer cages (1.2 m 0.8 m 0.5 m) and housed in an environmentally
controlled room maintained at 32 to 34C for the first week and then reduced by 2 to
3C per week to a final temperature of 20C. Broilers were allowed ad libitum access

to feed and water.

After fasting for 12 h, the birds of each replicate cage were weighed at 21 and 42
days of age. Feed intake was recorded during the 42-day trial. The average daily gain
(ADG), average daily feed intake (ADFI) and feed per unit gain (F/G) were
calculated.
At 21 and 42 days of age, one bird per replicate was randomly selected and
weighed following a 12-h fast. Blood samples were taken from jugular vein, collected
in plastic tubes and centrifuged for 15 min at 4500 g at 4C to obtain the blood
serum. Then, the broilers were killed by cervical dislocation and necropsied
immediately. The jejunal digesta samples were collected and stored at 20C until
analysis of enzyme activity.
From 17 to 19 and 38 to 40 days of age, excreta samples were collected from
each replicate. Feathers and shredded dry skin were removed carefully before
collection. The excreta samples were stored in sealed plastic bags and preserved
immediately in freezer (20C).
At 35 days of age, six male birds per treatment (one chicken per replicate) were
selected by similar body weight and allocated into metabolism cages (40 cm 40 cm
35 cm). Followed by a 32-h fast, the broilers were delivered their respective
experimental diet (60 g/each bird) by forced-feeding. Six hours after feeding, the
chickens were slaughtered by cervical dislocation. The total digesta was collected for
analyzing the amounts of digesta residue.
2.4. Analytical procedures
Pellet durability index (PDI) were determined according to the procedure of
Pfost (1963). In detail, pellet samples were first sieved on the appropriate sieve to
remove fines. Then the samples of sieved pellets (500 g) were tumbled in the

tumbling can device (THB30, Wuxi Taihu Grain Machinery Co., Ltd., Jiangsu, China).
After tumbling for 10 min at 50 r/min (Thomas and van der Poel, 1996), samples were
sieved and the amount of pellets not passing the sieve (diameter of 2 mm) was
determined. Pellet durability index was then calculated as the ratio of intact pellets
after tumbling to whole pellets at the start. Hardness was measured using 100 pellets
of each diet on a hardness tester (GWJ-1, Jiangsu Tongjun Instrument Technology Co.
Ltd., Jiangsu, China). Pellets, one at a time, were placed in the hardness tester and the
force needed to brake them was measured in kg.
The concentrations of glucose, serum urea nitrogen, total protein, albumin, total
cholesterol and triglyceride were determined by an automatic biochemistry analyzer
(VITALAB-Selectra-E,

Netherlands).

All

kits

were

provided

by

KeHua

Bioengineering Co., Ltd. (Shanghai, China).

The excreta and feed samples were dried for 48 h in an oven at 65C and then
allowed to equilibrate to atmospheric conditions for 24 h. After ground through a
0.45-mm screen, standard chemical analyses were carried out according to AOAC
(1990). Dry matter (DM) was analysed by drying samples to a constant weight at
105C (method 934.01). The ash content was determined after combustion at 550C
for 8 h (method 942.05). The organic matter (OM) was calculated as the weight loss
upon ashing. Nitrogen content was determined using the Kjeldahl technique with
CuSO4 as a catalyst (method 988.05) and crude protein (CP) was calculated as
N6.25. Ether extract (EE) was determined using HCl treatment prior to extraction
with petroleum ether in a continuous extractor (Soxhlet) (method 920.39).
Acid-insoluble ash (AIA) was determined using the method of Choct and Annison
(1992). The apparent digestibility of nutrients was calculated according to Zhou et al.
(2014).

The digesta samples of gastrointestinal tract were dried at 65 C for 48 h and

then at 105 C for 5 h in an oven. The dried samples were weighed after cooling down
to room temperature in a desiccator. The amount of digesta residue of gastrointestinal
tract was expressed as the weight (g) of the dried samples. The ratio of digesta residue
was calculated as the percentage (%) of amount of digesta residue to feed intake (dry
basis).
After thawing, the jejunal digesta samples were homogenized (1/9, w/v) with
ice-cold 0.9% sodium chloride solution using an Ultra-Turrax homogenizer (IKA,
Germany) and then centrifuged at 4500 g for 20 min at 4C. The supernatant was
collected and used for assaying enzyme activity. The activities of amylase, trypsin and
lipase as well as protein concentration were determined with commercial kits
(Jiancheng Bioengineering Institute, Nanjing, Jiangsu, China).
2.5. Statistical analysis
Data was analyzed using SPSS statistical software (Ver.16.0 for windows, SPSS
Inc., Chicago, IL, USA). The means and their pooled standard errors were presented.
Polynomial contrasts were used to test the linear and quadratic effects of palygorskite.
Significance was declared at P < 0.05.
3. Results
3.1. Feed technological characteristic
The findings showed that pellet quality of the starter and grower diets
significantly increased (linear and quadratic, P < 0.001, Table 4) as palygorskite
inclusion.
3.2. Growth performance
During starter period, the palygorskite supplementation improved ADG (linear
and quadratic, P < 0.001, Table 5) and ADFI (linear, P = 0.027 and quadratic, P =

0.007) of broilers. During grower and the whole period, the palygorskite
supplementation improved ADG and ADFI (linear and quadratic, P < 0.001). The
broilers fed the diet supplemented with 10 g/kg palygorskite had the highest ADG
among groups. Compared with the control group, neither 5 g/kg nor 10 g/kg
palygorskite inclusion affected feed/gain ratio (F/G) of broilers. However, the F/G in
grower period increased 1.6% with diet 15 g/kg (1.83 vs 1.80) and 3.3% with diet
20g/kg (1.86 vs 1.80).
3.3. Blood serum biochemical parameters
The palygorskite supplementation did not affect the concentrations of glucose,
serum urea nitrogen, total protein, albumin, globulin total cholesterol or triglyceride in
the blood serum (Table 6).
3.4. Apparent nutrient digestibility
The apparent digestibility of crude protein increased as palygorskite inclusion
during 17 to 19 d (quadratic, P = 0.025, Table 7).
3.5. The amount of digesta residue in the gastrointestinal tract
The amounts of digesta residue in the gastrointestinal tract was increased (linear
and quadratic, P < 0.001, Table 8) as palygorskite inclusion. The ratio of digesta
residue in the gastrointestinal tract was increased (linear and quadratic, P < 0.001).
3.6. Digestive enzyme activity in the jejunal digesta
No significant effect was found on the digestive enzyme activities in the jejunal
digesta with the palygorskite supplementation (Table 9).
4. Discussion
Clays can be incorporated as agglomerant in animal diets in order to improve
feed manufacture (Angulo and Brufau, 1995; Angulo et al., 1996; Pappas et al., 2010;
Zhang et al., 2015). In the present study, we found that dietary palygorskite inclusion

improved the hardness and durability index of pellets. Angulo et al. (1996) reported
that sepiolite, a clay with similar physical properties to palygorskite (Galn, 1996),
improved the performance of pelleted diets. Furthermore, the same authors reported
that sepiolite was more effective when fat and fiber levels were high (Angulo and
Brufau, 1995). Pappas et al. (2010) found that pellets manufactured with 1.0% added
palygorskite showed better pellet quality than those manufactured without
palygorskite. Improvement of pellet hardness and durability could be attributed to the
special sorptive and colloidal/rheological properties, which are the basis for its most
technological applications (Galn, 1996; Liu, 2007). In detail, the presence of
micropores and channels in palygorskite together with the fine particle size and
fibrous habit account for its very high surface area (Zhou, 2011; Xu et al., 2012).
Palygorskite may absorb polar liquid and form gel, which in turn may improve pellet
hardness and durability due to the increase of solid-solid bonding interaction.
Generally, clays have been incorporated in animal diets as a nonnutritive additive
to improve performance (Angulo and Brufau, 1995). However, the results of previous
experiments on the effect of clays on growth performance of birds are inconsistent.
Chalvatzi et al. (2014) found that the incorporation of 1.0% palygorskite in the laying
hen diets for 24 weeks improved laying percentage and feed conversion ratio. In
contrast, Qiao et al. (2015) found that dietary 2.0% palygorskite did not affect laying
performance of hens. Pappas et al. (2010) reported no significant effect of 1.0%
palygorskite supplementation on the live mass and feed conversion ratio of broilers.
Chen et al. (2016) and Cheng et al. (2016) reported neither 0.5% nor 1.0 %
palygorskite inclusion affected growth performance of broilers. In the present study,
palygorskite inclusion significantly enhanced ADFI and ADG during the overall
experimental period, but had no significant influence on feed conversion ratio. The

10

results in our study could be attributed to improved pellet quality by the addition of
palygorskite. Some studies indicated that broilers preferred hard to soft pellets,
resulting in an increase in feed intake (Jensen et al., 1962; Nir et al., 1994; Parsons et
al., 2006; Bouvarel et al., 2009). Vandegrift et al. (2003) reported that there was a
positive correlation between body weight and feed intake. Thus, the improved ADG
of broilers fed diets with palygorskite inclusion could be attributed to the increased
feed intake. It was notable that relatively high levels (20 g/kg) of dietary palygorskite
inclusion markedly reduced ADG compared with 1.0% and 1.5% palygorskite during
22-42 d and 1-42 d. This was in line with a previous study which reported that a lower
concentration of clay in the diet was more effective than a higher concentration (Oguz
and Kurtoglu, 2000). As indicated in the Table 2 and 3, the nutrients content in the
diets as organic matter, crude protein and ether extract were gradually reduced with
the increasing level of palygorskite inclusion. The adverse effects of higher
concentrations of palygorskite may be associated with the nutrition dilution and the
binding potential for nutrients aroused by palygorskite supplementation (Chalvatzi et
al., 2014; Zhang et al., 2015).
In the current study, palygorskite supplementation did no affect the blood serum
biochemical constituents of broilers. The obtained results in this study were in
agreement with the findings by Pappas et al. (2010) who reported that the
supplementation of 1.0% palygorskite in broiler diets did not affect hematocrit and
white blood cell values, as well as protein, albumin and globulin concentration in
blood. Moreover, the blood parameters of broilers fed a diet with 10% palygorskite
ranged within physiological values (Pappas et al., 2010).
Early works on the supplementation of clays in animal feed have found that
improvement of growth performance is also associated with improvements of nutrient

11

digestibility (Ouhida et al., 2000; Safaeikatouli et al., 2012). In the present study, the
addition of palygorskite inclusion tended to increase the nitrogen retention during
17-19 d. Safaeikatouli et al. (2012) explained that the improvement of nutrient
utilization may be attributed to the reduction of digesta passage rate. In our result,
palygorskite inclusion in the diets increased the amounts of digesta residue in
digestive tract of broilers, indicating the palygorskite could increase the digesta
retention time, which may contribute to enhanced nutrient utilization in our study.
It has been reported that 2.0% palygorskite inclusion in diets (mash) significantly
enhanced protease activity, but did not affect the activities of amylase, trypsin or
lipase in jejunal digesta in laying hens (Qiao et al., 2015). The actions of clay on
digestive enzymes have been reported that pancreatic enzymes can be adsorbed to the
surface of calys, forming complexes which are active over a range of different
digestive pH (Cabezas et al., 1991). In the current study, palygorskite inclusion in
diets (pellet) did not affect the digestive enzyme activities. The discrepancy may be
due to absorbing polar liquid by palygorskite and forming gel during pelleting process.
In this situation, the adsorptive capability for the digestive enzymes in gastrointestinal
tract was reduced accordingly.
5. Conclusions
In conclusion, our results suggested that palygorskite inclusion could enhance the
pellet hardness and durability index, and thus improve the average daily gain of
broilers. In additional, the optimal level of supplemented palygorskite in the pellet
diets of broilers was 10 g/kg.

12

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instead of zinc oxide on growth performance, apparent nutrient digestibility
and fecal zinc excretion in weaned piglets. J. Anim. Sci. 85, 435-439.

16

Thomas, M., van der poel, A.F.B., 1996. Physical quality of pelleted animal feed: 1
Criteria for pellet quality. Anim. Feed Sci. Technol. 61, 89112.
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nonlinear dynamics of daily broiler growth and feed intake. Poult. Sci. 82,
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Zhang, R.Q., Yang, X., Chen, Y.P., Yan, R., Wen, C., Liu, W.B., Zhou, Y.M., 2015.
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17

Table 1
Chemical analysis of palygorskite.
Chemical analysis (g/kg)
SiO2

563.2

Al2O3

102.5

MgO

133.4

CaO

101.2

Na2O

7.7

K 2O

11.5

Fe2O3

60.2

LOI1

20.2

Total

1000.0

LOI, loss on ignition.

18

Table 2
Composition and nutrient content (g/kg as fed) of the basal diets in starter period.
Items

Level of palygorskite (g/kg)

5
10
15

0
Ingredients
Corn
Soybean meal
Wheat middling
Corn gluten meal
Soybean oil
Limestone
Dicalcium phosphate
L-Lysine
DL-Methionine
Salt
Premix 1

492
330
50
44
35
11
20
3.7
1.3
3
10
0
1000

492
330
50
44
35
11
20
3.7
1.3
3
10
5
1005

492
330
50
44
35
11
20
3.7
1.3
3
10
10
1010

492
330
50
44
35
11
20
3.7
1.3
3
10
15
1015

20
492
330
50
44
35
11
20
3.7
1.3
3
10
20
1020

Palygorskite
Total2
Calculated analysis
AME (MJ/kg)
12.5
12.5
12.4
12.3
12.3
Crude protein
216
215
214
213
212
Calcium
10.0
9.95
9.90
9.85
9.80
Available phosphorus
4.70
4.68
4.65
4.63
4.61
Lysine
12.0
11.9
11.9
11.8
11.8
Methionine
5.00
4.98
4.95
4.93
4.90
Methionine +cysteine
8.40
8.36
8.32
8.28
8.24
Analyzed nutrient composition
Dry matter
900
900
898
896
890
Crude protein
223
222
220
219
219
Organic matter
851
846
839
835
824
Ether extract
47.9
47.3
46.9
46.1
45.7
Ash
49.6
53.7
59.2
61.1
66.1
1
Premix provided the following per kilogram of diet: vitamin A (transretinyl acetate),
10,000 IU; vitamin D3 (cholecalciferol), 3,000 IU; vitamin E (all-rac--tocopherol
acetate), 30 IU; menadione, 1.3 mg; thiamine, 2.2 mg; riboflavin, 8 mg; nicotinamide,
40 mg; choline chloride, 600 mg; calcium pantothenate, 10 mg; pyridoxineHCl, 4 mg;
biotin, 0.04 mg; folic acid, 1 mg; vitamin B12 (cobalamin), 0.013 mg; Fe (from ferrous
sulfate), 80 mg; Cu (from copper sulfate), 8 mg; Mn (from manganese sulfate), 110
mg; Zn (from zinc sulfate), 65 mg; I (from calcium iodate), 1.1 mg; Se (from sodium
selenite), 0.3 mg.
2
Total feed was 1000 g basal diet plus 0, 5, 10, 15 and 20 g of palygorskite,
respectively.

19

Table 3
Composition and nutrient content (g/kg as fed) of the basal diets in grower period.
Items

Level of palygorskite (g/kg)

5
10
15

0
Ingredients
Corn
Soybean meal
Wheat middling
Corn gluten meal
Soybean oil
Limestone
Dicalcium phosphate
L-Lysine
DL-Methionine
Salt
Premix 1

547
250
50
60
46
13
16
4.1
0.9
3
10
0
1000

547
250
50
60
46
13
16
4.1
0.9
3
10
5
1005

547
250
50
60
46
13
16
4.1
0.9
3
10
10
1010

547
250
50
60
46
13
16
4.1
0.9
3
10
15
1015

20
547
250
50
60
46
13
16
4.1
0.9
3
10
20
1020

Palygorskite
Total2
Calculated analysis
AME (MJ/kg)
13.1
13.0
13.0
12.9
12.8
Crude protein
196
195
194
193
192
Calcium
9.60
9.55
9.50
9.46
9.41
Available phosphorus
4.00
3.98
3.96
3.94
3.92
Lysine
10.5
10.5
10.4
10.3
10.3
Methionine
4.40
4.38
4.36
4.33
4.31
Methionine +cysteine
7.50
7.46
7.43
7.39
7.35
Analyzed nutrient composition
Dry matter
898
895
895
894
893
Crude protein
207
203
199
198
197
Organic matter
845
840
837
833
830
Ether extract
63.1
62.6
62.1
61.8
60.9
Ash
52.7
55.2
57.9
61.4
63.1
1
Premix provided the following per kilogram of diet: vitamin A (transretinyl acetate),
10,000 IU; vitamin D3 (cholecalciferol), 3,000 IU; vitamin E (all-rac--tocopherol
acetate), 30 IU; menadione, 1.3 mg; thiamine, 2.2 mg; riboflavin, 8 mg; nicotinamide,
40 mg; choline chloride, 600 mg; calcium pantothenate, 10 mg; pyridoxineHCl, 4 mg;
biotin, 0.04 mg; folic acid, 1 mg; vitamin B12 (cobalamin), 0.013 mg; Fe (from
ferrous sulfate), 80 mg; Cu (from copper sulfate), 8 mg; Mn (from manganese sulfate),
110 mg; Zn (from zinc sulfate), 65 mg; I (from calcium iodate), 1.1 mg; Se (from
sodium selenite), 0.3 mg.
2
Total feed was 1000 g basal diet plus 0, 5, 10, 15 and 20 g of palygorskite,
respectively.

20

Table 4
Influence of palygorskite on the pellet durability index (PDI) and pellet hardness of starter and grower diets.
Level of palygorskite (g/kg)

P-value

Items

SEM
0

10

15

20

Hardness (kg)

1.51

3.13

4.02

4.45

4.97

PDI (%)

93.1

94.0

96.2

96.1

Hardness (kg)

1.50

1.64

2.97

PDI (%)

92.8

93.3

94.4

Linear

Quadratic

0.281

<0.001

<0.001

97.2

0.399

<0.001

<0.001

3.60

4.04

0.236

<0.001

<0.001

94.7

97.7

0.410

<0.001

<0.001

Starter diet

Grower diet

SEM, total standard error of means.

a-e

Means within a row with different superscripts are different at P < 0.05.
Table 5
Influence of palygorskite on the average daily gain, average daily feed intake and feed per unit gain of broilers.
Items

Level of palygorskite (g/kg)

0

10

21

15

20

SEM1

P-value
Linear

Quadratic

Starter period
Average daily gain (g/bird)

34.1

36.4

37.2

37.0

36.8

0.229

<0.001

<0.001

Average daily feed intake (g/bird)

46.5

49.6

50.6

49.7

49.8

0.437

0.027

0.007

Feed per unit gain (g feed/g gain)

1.37

1.36

1.36

1.34

1.35

0.00805

0.424

0.715

Average daily gain (g/bird)

80.9

86.3

92.2

90.8

87.9

0.746

<0.001

<0.001

Average daily feed intake (g/bird)

146

155

166

166

163

1.66

<0.001

<0.001

Feed per unit gain (g feed/g gain)

1.80

1.79

1.80

1.83

1.86

0.0115

0.060

0.093

Average daily gain (g/bird)

57.5

61.4

64.7

63.9

62.4

0.464

<0.001

<0.001

Average daily feed intake (g/bird)

96.1

102

108

108

107

0.964

<0.001

<0.001

Feed per unit gain (g feed/g gain)

1.67

1.66

1.67

1.69

1.71

0.00904

0.097

0.162

Grower period

Whole trial period

SEM, total standard error of means.

a-c

Means within a row with different superscripts are different at P < 0.05.
Table 6
Influence of palygorskite on the serum biochemical indices of broilers.
Level of palygorskite (g/kg)
Items
0
5
10
15

22

20

SEM1

P-value
Linear

Quadratic

21d
Glucose (mmol/L)

13.4

13.7

13.2

13.3

12.8

0.142

0.121

0.196

Serum urea nitrogen (mmol/L)

0.635

0.658

0.658

0.703

0.710

0.0141

0.048

0.147

Total protein (g/L)

28.7

29.6

29.6

30.9

29.3

0.357

0.329

0.266

Albumin (g/L)

16.7

17.0

16.8

16.7

16.8

0.187

0.930

0.985

Globulin (g/L)

11.9

12.7

12.8

14.2

12.5

0.292

0.209

0.144

Total cholesterol (mmol/L)

3.51

3.48

3.34

3.36

3.19

0.0613

0.075

0.207

Triglyceride (mmol/L)

0.507

0.570

0.490

0.472

0.493

0.0146

0.231

0.487

Glucose (mmol/L)

12.8

13.4

13.6

13.2

13.5

0.220

0.444

0.642

Serum urea nitrogen (mmol/L)

0.678

0.647

0.625

0.633

0.640

0.0130

0.354

0.422

Total protein (g/L)

32.7

34.0

32.7

34.7

35.3

0.524

0.114

0.265

Albumin (g/L)

16.1

16.2

16.9

17.7

17.3

0.318

0.077

0.206

Globulin (g/L)

16.6

17.8

15.9

17.0

18.0

0.503

0.598

0.715

Total cholesterol (mmol/L)

2.89

2.65

2.82

2.71

2.96

0.0584

0.619

0.335

Triglyceride (mmol/L)
0.505
0.422
0.478
0.508
1
SEM, total standard error of means.
Table 7
Influence of palygorskite on the apparent digestibility of nutrients in broilers.

0.533

0.0202

0.325

0.326

42d

Items

Level of palygorskite (g/kg)

23

SEM1

P-value

10

15

20

Linear

Quadratic

Organic matter (%)

74.0

75.6

75.2

73.8

72.8

0.457

0.194

0.114

Crude protein (%)

57.8

61.7

61.7

60.9

60.5

0.491

0.198

0.025

Ether extract (%)

69.8

71.8

69.1

69.0

69.2

0.596

0.348

0.638

Organic matter (%)

74.3

76.5

75.1

74.6

74.6

0.502

0.748

0.688

Crude protein (%)

59.0

62.9

62.4

62.2

62.6

0.620

0.137

0.142

Ether extract (%)

73.3

74.9

73.8

73.8

72.9

0.348

0.467

0.352

17-19 d

38-40 d

SEM, total standard error of means.

a-b

Means within a row with different superscripts are different at P < 0.05.

24

Table 8
Influence of palygorskite on the amounts and ratio of digesta residue in the gastrointestinal tract.
Level of palygorskite (g/kg)

Items

P-value

SEM1

10

15

20

Amounts of digesta residue (g)

31.6

35.3

43.8

46.1

47.8

0.657

<0.001

<0.001

Ratio of digesta residue (%)

58.7

65.6

81.7

85.9

89.3

2.94

<0.001

<0.001

Linear

Quadratic

SEM, total standard error of means.

a-d

Means within a row with different superscripts are different at P < 0.05.

Table 9
Influence of palygorskite on the intestinal enzymes activities of broilers.
Level of palygorskite (g/kg)

P-value
SEM1

Items
0

10

15

20

Linear

Quadratic

Amylase (IU/mg prot)

52.6

54.8

53.5

53.7

52.1

3.80

0.880

0.825

Trypsin (IU/g prot)

7.76

7.54

7.43

7.38

7.61

0.427

0.959

0.839

21d

25

Lipase (IU/mg prot)

47.5

48.1

47.0

46.7

48.0

4.54

0.910

0.993

Amylase (IU/mg prot)

50.9

53.2

53.8

53.6

51.8

5.26

0.829

0.269

Trypsin (IU/g prot)

7.66

7.92

7.65

7.65

7.34

0.661

0.687

0.923

Lipase (IU/mg prot)

39.8

40.4

38.1

38.0

39.0

2.77

0.689

0.546

42d

SEM, total standard error of means.

26