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International Journal of Agriculture and Crop Sciences.

Available online at www.ijagcs.com


IJACS/2013/6-18/1239-1245
ISSN 2227-670X 2013 IJACS Journal

Various Physico-chemical Properties of Dietary Fiber


Sources of Poultry Diets
H.R. Hemati Matin1, F. Shariatmadari*2, M.A. Karimi Torshizi3
School of Agriculture, Tarbiat Modares University, Tehran-Iran, 14115-336

*Corresponding author email: shariatf@modares.ac.ir, shariat_madari@yahoo.com

ABSTRACT: This study was carry out to investigated the proximate composition together with some
water and oil functional characteristics of wheat bran (WB), barley bran (BB), and soybean hulls (SH) as
poultry dietary fiber (DF) sources. The results showed that moisture contents (range 6.04 to 7.71%) were
similar between tested DF sources. The SH had notably higher crude protein content rather that of WB
and BB (P<0.05). The ash content of SH and BB did not significantly differ but were higher than that of
WB (P<0.05). The order of DF ether extract was SH> BB > WW (P<0.05). The highest and lowest
amounts of crude fiber, and acid detergent fiber were related to BB and WB, respectively (P<0.05).
Moreover, BB was dedicated the highest level of neutral detergent fiber, acid detergent lignin, bulk
density, cation-exchange capacity, and water holding capacity rather other DF sources. The pH of all
three DF was slightly acidic (range from 5.25 to 6.00), although their pH did not significantly differ. The BB
and SH viscosity were statically similar but both were higher than WB viscosity (P<0.05). The order of
water insoluble solids of DF sources was as following BB>WB>SH (P<0.05). No significantly differences
were found between the swelling capacity of BB and SH but both were higher than that of WB (P<0.05).
The oil holding capacity of DF sources for the tested oils (corn, olive oil and melted tallow) was in order
SH>BB>WB, respectively (P<0.05). The emulsifying activity of the tested DF sources for corn and olive
oils was similar. The measured emulsion stability for corn oil was similar between SH and BB but both
were higher than ES of WB. The results of current study indicated that the physico-chemical properties of
various tested DF sources are dependent on their composition, especially fiber content. These data
provide a clear insight into how fiber fraction of poultry diets interacts with water and oil and how the
potential nutritional consequences were determined in terms of physico-chemical properties.
Keywords: Barley bran; poultry diet; soybean hulls; wheat bran
Abbreviations: ADF-Acid Detergent Fiber; ADL-Acid Detergent Lignin; BB-Barley Bran; BD-Bulk Density;
CEC-Cation-exchange Capacity; CF-Crude Fiber; CP-Crude Protein; DF-Dietary Fiber; DM-Dry Matter;
EA-Emulsifying Activity; ES-Emulsion Stability; EE-Ether Extract; GIT-Gastrointestinal Tract; NDF-Neutral
Detergent Fiber; OHC-Oil Holding Capacity; SH-Soybean Hulls; SC-Swelling Capacity; TDS-Total
Dissolved Solute; WHC-Water Holding Capacity; WIS-Water-insoluble Solids; WB-Wheat Bran

INTRODUCTION

Dietary fiber (DF) is an important component of poultry diets. The inclusion of suitable level of DF in poultry
diets and the recognition of DF as a feed substance with significant nutritional value has increased dramatically
over the past 40 years (Owusu-Asiedu et al., 2006; Mateos et al., 2012). Dietary fiber defines as cell walls of plant
tissues (McDonald et al., 2002) that are resistant to digestion, absorption in gastrointestinal tract (GIT), and are
heterogeneous in terms of nutritional, chemical and physical nature (Sarikhan et al., 2010; Mateos et al., 2012). It is
approved that amount and type of DF affect the feature of the GIT functions and thereby poultry production
(Owusu-Asiedu et al., 2006; Jimnez-Moreno et al., 2009; Saki et al., 2010). On the other hand, the increased
awareness of the potential health benefits of DF has undoubtedly encouraged nutritionists to recognize in more
details DF sources properties. Little scientific literature exists that investigated the physico-chemical properties of
the DF sources. It seems more comprehensive information needs regarding the characteristics of DF sources.
Therefore, in the present study, wheat bran, barley bran as well as soybean hulls were selected as a poultry DF
sources and their characteristics (proximate analysis, water and oil related specification) were evaluated and
compared.
Intl J Agri Crop Sci. Vol., 6 (18), 1239-1245, 2013

MATERIALS AND METHODS

Dietary Fiber Samples


Wheat bran (WB), barley bran (BB) and soybean hulls (SH) were used as representation of DF sources in
poultry diets which purchased from the Central Cooperative Poultry, Tarkhineh Company and Shayan Protein and
Energy Company (Tehran, 2013, Iran), respectively. The milled DF sources powders were transferred to airtight
plastic bags and stored in a desiccator at room temperature (24C) prior to parameters determination.

Proximate Analysis
The chemical compositions of DF sources were determined based on procedures described by the AOAC
(1995) methods with some modifications. Briefly, the moisture was determined as the weight loss for 16 h at 105C
followed 2 h at 130C at air-oven drying (Debon and Tester, 2001). The crude protein (CP) was estimated by
TM
multiplying the nitrogen content, determined by a Kjeltec (Foss, 8400), with a factor of 6.25. The ash content was
determined in a muffle furnace by slowly heating 2 g of each sample up to 500C for 16 h (Debon and Tester,
2001). The ether extract (EE) was ascertained using a Soxhlet apparatus with extra petroleum benzine. The crude
fiber (CF) was measured after samples were boiling for 30 minutes in each sulfuric acid (1.25%) and sodium
TM
hydroxide (1.25%) using Fibertec (Foss, 2010). Moreover, the neutral detergent fiber (NDF), acid detergent fiber
(ADF) and acid detergent lignin (ADL) were determined based on Van Soest et al (1991) methods.

Physico-chemical Properties
The bulk density (BD) was measured using Wang and Kinsella (1976) method with slight modification. A
pre-weighed graduated cylinder (50 mL) was filled to 50 mL with each DF samples, followed by packing with gently
tapping the cylinder on the bench-top until the volume was constant. The BD was recorded as a ration of weight (g)
of DF sample to its volume (mL).
The cation-exchange capacity (CEC) was determined according to Moorman et al (1983) procedure. In
brief, the cationic functional groups of sample (500 mg) were first converted into their acidic form by stirring in 25
mL of 2 M HCl overnight. After centrifugation (15 min, 2,500 g, 25C), the residue was washed extensively with
ultrapure water until the total dissolved solute (TDS) of the washing water was similar to that of the ultrapure water
(<5.0 mg/L). Subsequently, the acidic residue was suspended in 25 mL of 0.3 M NaCl and was centrifuged under
the same above condition. The supernatant was then titrated with 0.01 M KOH, and the CEC was expressed as
milliequivalents per kg (meq/kg) of DF. The oil holding capacity (OHC) of all DF samples was determined by the
method of Caprez et al (1986). About 2 g of each sample was placed in a 50 mL graduated centrifuge tube,
followed by the addition of 20 mL of each tested oils (corn oil, olive oil, melted tallow). The tube was left for 30 min
at room temperature (25C) with agitation. Subsequently, the mixture was centrifuged (2,500 g, 25C, 30 min) and
the oil supernatant was decanted and weighed. The OHC of the individual DF samples was expressed as gram of
oil held by one gram of corresponding DF samples. The emulsifying activity (EA) and emulsion stability (ES) of the
DF samples were determined according to Yasutmasu et al (1972) method. Briefly, one gram of each DF samples
was weighed into 50 mL graduated centrifuge tube. Then, 12.5 mL of ultrapure water was added and mixture was
homogenized at 12,000 rpm for 30 second (WTW-DISPER D-8). An aliquot (12.5 mL) of each tested oils (corn oil,
olive oil, melted tallow) were then added into the slurry and further homogenized 12,000 rpm for 60 second. The
emulsion formed was centrifuged (1,200 rpm, 25C, 5 min) and the EA was calculated from the ration of the depth
of the emulsified layer to the depth of the total volume of content inside the centrifuge tube (as a percentage). For
determining ES, the emulsion formed was further heated in a 80C water bath for 30 min and then cooled down to
room temperature (25C) followed by centrifugation under the above same conditions. The ES was calculated in
the same way as EA and also expressed as percentage of the unheated control.
The pH values of 0.1% (w/v) DF suspensions were determined at 25C with a calibrated pH meter
(Metrohm, Germany) (Debon and Tester, 2001). The viscosities of 0.1% (w/v) aqueous were determined at 25C
by a viscometer (DV-II LV, Brookfield, Stroughton, MA, USA) previously calibrated with a silicone standard. The
water holding capacity (WHC) of DF samples was measured by the modified centrifugation methods described by
Suzuki et al (1996). In brief, 20 mL of ultrapure water was added into a centrifuge tube containing 200 mg of DF
samples. Subsequently, the centrifuge tubes were incubated in a 25C shaking water bath for 24 h. After
centrifuged (14,000 g for 30 min at 25C) (Hettich Zentrifugen D-78532), the supernatant was discarded and the
moisture content of pre-weighed pellet was determined after dehydration in an air oven for 2 h at 120C. The WHC
of each DF sample was expressed as the weight of water held by one gram of corresponding DF samples. The
water-insoluble solids (WIS) was determined following the method of Massiot and Renard (1997) with slight
modifications and based on the method of Huang et al (2011). The samples were homogenized in cold ultrapure

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water for 1 min (DF-to-water ratio of 1:30, w/v). After filtration, the residue was washed with 70% ethanol, dried by
solvent-exchange and air-oven at 40C for 48 h. Then, the content of WIS was corrected for residual protein and
ash contents. The swelling capacity (SC) of DF was measured based on the method of Gmez-Ordez et al
(2010). Briefly, dry powder samples (500 mg) of DF sources were weighed in a 10 mL measuring cylinder (0.1 mL
graduations) and 10 mL ultrapure water was added. Then, it was stirred gently to eliminate trapped air bubbles and
left on a level surface at room temperature overnight (18 h) to allow sample to settle. The volume (mL) occupied by
the samples was measured and SC was expressed as mL/g of dry sample.

Statistical Analysis
All data were analysed for normal distribution using the NORMAL option of the UNIVARIATE procedure
and for homogeneity of variances for treatment means through the Levenes Test, using the HOVTEST option of
GLM procedure of SAS (2004). All determinations were performed at least in triplicate and were analysed as a
completely randomized design by the GLM procedure of SAS (2004). Significant differences were compared by
Tukey tests (P<0.05). All differences were considered significant at P0.05.

RESULTS AND DISCUSSION

The Analysis of Chemical Composition


Table 1 shows the proximate composition of different tested DF sources. Narrow range of moisture (6.04
to7.71%) cause to have no significant differences in dry matter (DM). As explained above, the moisture content
was calculated as weight losses at 105C overnight and 130C for 2 h. It is showed that this method would result in
accurate estimation rather than weight losses at 105C overnight (Debon and Tester, 2001). Differences in
moisture contents might be due to processing or storage condition (Idouraine et al., 1996). It seems neither
processing and nor storage condition did not influence tested DF sources moisture in this study. Moreover, the
calculated moisture content is <10%, which are in agreement with the results of other studies for various DF
sources (Hassani, 1989; Idouraine et al., 1996).
The results indicated that SH had significantly higher CP content rather WB and BB (P<0.05). In the
present study, the CP content of SH (25.26%) is much higher than other studies (9 to 10%: Hassani, 1989; 11.1%:
Chee et al., 2005; 14%: Ward and Reichert, 1985). The presence of whole and crushed the grains of soybean
grains in SH which used in current experiment seems be the most important reason for the difference in CP
contents. A range of 9.2 to 18.7% for CP was reported from different sources for SH (Cole et al., 1999). Crop
varieties, sample size, and preparation methods are effective in protein contents (Hassani, 1989). The measured
CP content for WB (14.05%) fall in considerable range with other studies (11.8 to 14.5%: Pomeranz, 1988; 14%:
Dreher, 1987; 6.8 to 16.88%: Hassani, 1989; 15.4%: Chee et al., 2005; 12.6 to 20.0%: Peterson et al., 1983).
No significantly difference was found among the ash content of SH and BB but both was higher than that of
WB (P<0.05). The range (4.59 to 6.93%) ash contact measured for DF sources in this experiment show that their
ash contents are low and have small proportion. The obtained values for WB (4.59%) and for SH (6.46%) in this
study are analogous with other researches (WB, 6.08%; SH, 4.32%; Hassani, 1989; WB, 6.20 to 11%; Pomeranz,
1988; and WB, 5.1%, SH, 4.9%; Chee et al., 2005). Differences in soil are the main reason for variation in ash
contents of crop products. Generally, because of systematic data on the ash composition of DF sources are limited
the comparing of different studies is difficult.
The highest EE of DF sources was in order the SH >BB >WW (P<0.05). Based on reported EE values of
oat bran (9.5%: Youngs, 1986) and wheat (3.6 to 5.6%: Pomeranz, 1988; 3.6%: Chee et al., 2005) obtained results
in current study for WB (3.68%) are consistency but for SH is inconsistency (1.5%: Chee et al., 2005). Differences
in EE might be related to the methods of measurement, type of solvent, and variety differences.
The highest and lowest amounts of CF were related to BB and WB, respectively (P<0.05). The amount of
measured DF for WB (8.25%) is in agreement with other research (8.7%) but it is lower for SH (13.97% vs. 32.5%)
(Chee et al., 2005). This inconsistency for SH can be attributed to presence of whole grain in used SH in this study.
The highest and lowest ADF content were related to BB and WB, respectively (P<0.05). The measured ADF values
for WB (11.22%) fall in the reported range of values in other researches (11.48 to 15.86%: Hassani, 1989; and
10.7%: Chee et al., 2005) but is lower that other study (Idouraine et al., 1995). The ADF results of SH (18.95%)
and BB (32.21%) is opposed with others (SH, 39.48 to 46.00%: Hassani, 1989; 36.80%: Chee et al., 2005; 48.82%:
Idouraine et al., 1995; and BB, 24.76%: Idouraine et al., 1995). The highest level of NDF was dedicated to BB
(P<0.05). Moreover, the highest amount of measured lignin attributed to BB (P<0.05). The measured NDF value for
WB is slightly less than, but close to, reported values (41.0%: Chee et al., 2005) but, for SH (26.00%) is much
lower than reported values (52.9%: Chee et al., 2005). Finally, it must be noted that differences in the amount of

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CF and its components can be depends on the conditions of separated and proportion of products, environmental
conditions, DF varieties, and intrinsic characteristics of DF sources.
Furthermore, Table 1 illustrated that the BD of BB was significantly higher than that of the DF sources
(P<0.05). The variation of BD between three DF sources would mainly depend on their differences in particle
density as well as interstitial air volume during processing and preparation conditions (Singh et al., 2003). However,
the very porous structure of the BB seems let to occupy with higher air which resulted in lower BD compared to that
of other tested DF sources.
The highest measured CEC order was attained to BB>WB>SH (P<0.05). The CEC are depended on the
sulfate, acid uronice content, and charge density (Carvalho et al., 2009). Despite significantly differences, CEC was
considerably low in tested DF sources (9.6 to 29.9 meq/kg). The results are considerable with cellulose values in
other researches (22.7 to 23 meq/kg: Chau and Huang, 2003; 2004); 19.0 meq/kg: Thompson and Weber, 1981).
Finally, the low CEC of the various DF sources could attribute to low acid uronice content (Wong et al., 2005).

Physico-chemical Functional Properties


Table 2 shows the measured water related characteristics of the tested DF sources. The pH of all three DF
sources was slightly acidic (ranged from 5.25 to 6.00). The results suggest that their incorporation in a diet system
might decrease final pH of diet. The difference in DF sources pH influences fiber solubility. This difference in fiber
solubility could affect other chemical properties of the DF sources (e.g., viscosity) and influence their ability to react
with other nutrients in GIT (Luccia and Kunkel, 2002). Little information is available about pH of DF source
suspensions to allow useful comparisons.
No statistical differences were observed between viscosity of BB and SH but both were higher than WB
viscosity (P<0.05). The viscosity of DF source suspensions are depends on the solubility, molecular weight, and
structural chemical bonds of the DF sources (Choct, 2002). Moreover, the molecular size, spatial shape, and
concentration of the DF sources affect DF viscosity. The solubility, molecular weight, spatial shape, and structural
linkages might have caused the differences in DF viscosity in this study. In fact, the spatial structure of BB and SH
have the water absorption and gel formation characteristics and can cause increased viscosity of the prepared
suspensions.
The highest WHC of DF sources was related to BB source (P<0.05). The order of WIS of DF sources was
as BB>WB> SH (P<0.05). No significantly differences were observed between the SC of BB and SH but both of
them were higher than WB. Associated water is important in emerge of DF sources properties. Such water would
affect DF sources metabolic activity across with GIT. Water soluble and insoluble portions of DF sources have
critical roles in WHC, WIS, and SC of DF sources. It showed that DF sources with high insoluble portion induce
lower WHC and SC and higher WIS (Jimnez-Moreno et al., 2009; Gmez-Ordez et al., 2010; Mossami et al.,
2011).
As shown in Table 2, SC and WHC of DF sources follow a same trend as higher CF content led to higher
SC and WHC. It demonstrated that WHC of DF sources is attributed to insoluble fiber, while other ascribed it to
high acid uronice content (Femenia et al., 1997; Ruprez and Saura-Calixto, 2001). However, there is general
agreement that this character depends on experimental conditions (temperature, pH, time, and characteristics of
centrifuge), particle size and processing (Michel et al., 1988; Suzuki et al., 1996; Femenia et al., 1997). The WHC
of a DF sources measures the amount of water retained by the fiber after subject to a stress such as centrifugation
(Nelson, 2001). It is an important hydration property of a DF source that needs to be measured before their
incorporation on the diets. The results showed that BB exhibited high value of WHC than that of other DF sources
(P<0.05). In the present study, however, the values obtained for DF sources were lower than that of some DF
cereal byproducts (WB: 6.40 to 6.60 g/g, Adams et al., 1986; Ralet, 1990; oat bran: 5.50 g/g, Cadden, 1987) as
well as some commercial DF-rich supplements (AGIOLAX: 6.60 g/g; Goni and Martin-Carron, 1998), but higher
than that of wheat (2.6 g/g), corn (2.5 g/g) and soy bran (2.4 g/g) (Dreher, 1987). The WHC determined by
centrifugation method in the present study represented all three types of waters (water bound to the hydrophilic
polysaccharides, held within the fiber matrix, trapped within the cell-wall lumen) associated with the fibers (Fleury
and Lahaye, 1991). Numerous factors could influence the DF water related parameters. Although raw sources of
DF is determinant factor for its chemical composition and structure, also, final microstructure (fiber length, particle
size and porosity) and processing conditions are important (Nelson, 2001). The microstructure of DF is believed to
have more profound effect that its chemical composition on the WHC (Robertson and Eastwood, 1981). In the
present study, differences in WHC between three DF source samples (be higher in BB) might be due to their
variation in particle size. Since the smaller the particle size of water insoluble portions, the larger surface area for
exposure of their hydrophobic regions would be results which might cause to lower WHC value. However, it is
understood that reduction in WB particle size decreased its WHC due to the loss of insoluble fiber (Cadden et al.,

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1987). This observation emphasizes the importance of DF structure with water related parameters (Esposito at al.,
2005). Furthermore, the methods of measurement and DF nature (pH, ionic strength, presence of other functional
materials) are also crucial important factors which affect the WHC, WIS and SC of DF.
The measured oil related parameters of tested DF are shows at Table 3. The OHC for the tested oils (corn,
olive oil and melted tallow) showed that high ability was related to SH>BB>WB, respectively (P<0.05). The OHC is
another physico-chemical property of a high fiber ingredient that can be applied in the formulated diets. In the
present study, only OHC of BB for corn and olive oils (1.79 and 1.81 g/g, respectively) was comparable to that of
wheat DF (2.3 g/g) (Thebaudin et al., 1997). Although the OHC values (corn and olive) for WB was notably low

(1.21 and 1.31 g/g, respectively), they were comparable to that a FIBREX (1.29 g/g) (Abdul-Hamid and Luan,
2000). The ability of a fiber to bind oil is more of a function of that porosity of fiber structure than the affinity of the
fiber molecule for oil (Nelson, 2001). Therefore, pre-processing of DF with water and reduction of particle size
would result in lower OHC value (Thebaudin et al., 1997; Nelson, 2001), since the number of pores in fiber is
reduced or the pores are pre-occupied by water. However, it seems that a more complex mechanism determining
that variations of OHC of DF might be involved such as number of lipophilic sites, overall hydropoboicity and
capillary attraction (Kinsella, 1976). The DF with a high OHC allows the stabilization of high fat content and
emulsion by retaining the fat in formulated diet. On the other hand, the high OHC might interference with lipid
absorption in GIT (Carvalho et al., 2009).
No significantly differences were observed among the EA of the tested DF for corn and olive oils. The
measured corn oil ES, also, showed that no differences had between SH and BB but both was higher than ES of
WB (P<0.05). Also, the highest and lowest olive oil ES were related to BB and WB, respectively (P<0.05). The
results showed that the emulsion formed by DF (for corn and olive oils), except BB, was undesirable, since their EA
values (27.00 to 38.84%) were <50% (Wang and Kinsella, 1976; Yasutmasu et al., 1972). These findings are in

compares with rice bran (14.4%) and FIBREX (3.46%) (Abdul-Hamid and Luan, 2000). The notably low values of
the EA of tested DF are suggesting their poor potential to act as an emulsifier in formulated diets. Furthermore,
despite significant differences in the ES, emulsion formed by all DF were very stable which was evident by their
similarly high percentage of ES after incubation at 80C water bath for 30 min.

Table 1. Chemical composition, bulk density and cation-exchange capacity of tested dietary fibers (dmb)
Treatment DM Pr EE Ash CF NDF ADF ADL BD CEC
Wheat bran 92.29 14.05b 3.68b 4.59b 8.25b 31.73ab 11.22c 8.63c 0.48a 16.90b
Soybean hulls 93.88 25.26a 12.07a 6.46a 13.97ab 26.00b 18.95b 12.28b 0.42a 9.60c
Barley bran 93.96 8.10c 4.19ab 6.93a 21.39a 54.83a 32.21a 14.64a 0.25b 29.90a
SEM 3.202 3.202 1.717 0.456 2.476 5.641 3.889 1.119 0.045 3.763
P-value 0.47 0.002 <0.001 0.002 0.013 0.004 0.001 0.003 0.013 0.003
DM: Dry Matter; Pr: Protein; EE: Ether Extract; CF: Crude Fiber; NDF: Neutral Detergent Fiber; ADF: Acid Detergent Fiber;
ADL: Acid Detergent Lignin; BD: Bulk Density (g/mL); CEC: Cation-Exchange Capacity (meq/kg); Means with different
superscripts in same column are significantly different (P<0.05).

Table 2. Some water related properties of tested dietary fibers


Treatment pH Viscosity (cP) WHC (g/g) WIS SC (mL/g)
Wheat bran 5.26 1.74b 3.41b 61.55b 5.75b
Soybean hulls 5.99 1.93a 3.91b 52.41c 7.35a
Barley bran 6.00 2.00a 4.57a 68.19a 8.00a
SEM 0.208 0.050 0.217 2.917 0.448
P-value 0.30 0.017 0.009 0.002 0.037
WHC: Water Holding Capacity; WIS: Water Insoluble Solids (g/100 g DM corrected for nitrogen and ash); SC: Swelling
Capacity; Means with different superscripts in same column are significantly different (P<0.05).

Table 3. Some oil related properties of tested dietary fibers


Treatment OHCC (g/g) OHCO (g/g) OHCT (g/g) EAC (%) EAO (%) ESC (%) ESO (%)
Wheat bran 1.21c 1.31c 2.83c 32.84 27.00 81.92b 92.89b
Soybean hulls 2.77a 2.90a 4.25a 29.50 28.39 96.08a 96.03ab
Barley bran 1.79b 1.81b 3.16b 52.00 49.72 95.86a 96.66a
SEM 0.288 0.297 0.271 5.505 5.573 3.126 0.777
P Value 0.001 0.001 0.001 0.21 0.17 0.33 0.031
OHCC: Oil Holding Capacity for Corn; OHCO: Oil Holding Capacity for Olive; OHCT: Oil Holding Capacity for Tallow; EAC:
Emulsifying Activity for Corn; EAO: Emulsifying Activity for Olive; ESC: Emulsion Stability for Corn; ESO: Emulsion Stability for
Olive; Means with different superscripts in same column are significantly different (P<0.05).

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CONCLUSION

Chemical and physical properties of tested dietary fibers which would provide a clue to its physiological
function will helpful for their selection to including in poultry diets. Tested dietary fiber sources could be as a good
source of dietary fiber with multiple functional tasks and significant technical advantages. The results of current
study indicated that DF components would likely be the main determining factors for physic-chemical and functional
properties assessed in this trial.

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