Archives of Animal Nutrition

ISSN: 1745-039X (Print) 1477-2817 (Online) Journal homepage: http://www.tandfonline.com/loi/gaan20

Effects of glyceryl polyethylene glycol ricinoleate

on nutrient utilisation and performance of broiler
chickens

Sebastian Andrzej Kaczmarek, Maciej Bochenek, Anne-Cathrine Samuelsson

& Andrzej Rutkowski

To cite this article: Sebastian Andrzej Kaczmarek, Maciej Bochenek, Anne-Cathrine

Samuelsson & Andrzej Rutkowski (2015) Effects of glyceryl polyethylene glycol ricinoleate on
nutrient utilisation and performance of broiler chickens, Archives of Animal Nutrition, 69:4,
285-296, DOI: 10.1080/1745039X.2015.1061722

To link to this article: http://dx.doi.org/10.1080/1745039X.2015.1061722

Published online: 06 Jul 2015.

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 Archives of Animal Nutrition, 2015
Vol. 69, No. 4, 285–296, http://dx.doi.org/10.1080/1745039X.2015.1061722

Effects of glyceryl polyethylene glycol ricinoleate on nutrient

utilisation and performance of broiler chickens
a
Sebastian Andrzej Kaczmarek *, Maciej Bochenekb, Anne-Cathrine Samuelssonc and
Andrzej Rutkowskia
a
Department of Animal Nutrition and Feed Management, University of Life Sciences, Poznan, Poland;
b
Biochem Polska Sp. z o.o., Toruń, Poland; cAkzoNobel SC AB, Stenungsund, Sweden
Downloaded by [Florida Atlantic University] at 02:37 06 November 2015

(Received 18 February 2015; accepted 9 June 2015)

A completely randomised design study with a 2 × 2 factorial arrangement was

conducted to observe effects of an emulsifier (glyceryl polyethylene glycol ricinoleate
(GPR)) on nutrient utilisation and performance of broiler chickens. A total of 384 male
broiler chickens were used to determine the influence of GPR (without addition or
added at 0.04% of diet) and two levels of apparent metabolisable energy (AMEN)
(according to standard requirements (Diets SE) or energy reduced by 0.4 MJ/kg diet
(Diets LE)) on birds’ performance and apparent total tract digestibility (ATTD). ATTD
of ether extract (EE) and AMEN were measured on d 14 and 35, and ATTD of neutral
detergent fibre (NDF) was measured on d 35. All diets were based on wheat–maize–
soybean meal with rapeseed oil and lard (blend of 50:50) as fat sources. During the
grower period and the whole trial, birds fed Diets supplemented with GPR were
characterised by higher body weight gain (BWG) and lower feed conversion ratio
(FCR) compared to chicken receiving diets without GPR (p < 0.05). At the end of
experiment, birds fed Diet LE without GPR were characterised by lower BWG and
higher FCR (p < 0.05). Supplementation with GPR caused a higher ATTD of EE for
diets SE and LE at d 14 and 35 (p < 0.05). Moreover, the GPR addition to Diet LE
improved ATTD of NDF at d 35 (p < 0.05). In conclusion, the findings suggest that
GPR effects the digestion of a blend of animal fat/rapeseed oil positively, even in a
practical maize–wheat-based broiler diet with decreased AMEN level.
Keywords: broilers; digestibility; emulsifiers; fat; nutrient availability; performance

1. Introduction
To increase the energy concentration in broiler chicken diets, usually animal fats and
vegetable oils are added. In consequence, the growth performance of birds can be
improved. According to Noy and Sklan (1995), lipase secretion in young birds is
insufficient to digest fats and oils properly, but Al-Marzooqi and Leeson (1999) reported
that exogenous lipase had no effect on digestion of fats and birds’ growth performance.
Under normal physiological conditions, the gastrointestinal tract of a bird is an aqueous
environment. Fatty acids as hydrophobic components have to aggregate to form micelles
to get absorbed. Emulsifiers, such as bile salts, naturally mediate this process. According
to Noy and Sklan (1995) and Uni et al. (1999), the limited digestion of dietary fats in
young birds is due to a reduced secretion capacity of bile salts and lipase until the end of
second week of life. It could be assumed that addition of bile salts or exogenous
emulsifiers to broiler diets could affect fat emulsification and, consequently, absorption

*Corresponding author. Email: sebak1@up.poznan.pl

© 2015 Taylor & Francis

 286 S. Andrzej Kaczmarek et al.

positively. Available research data regarding effectiveness of exogenous emulsifiers are

limited and inconsistent. Polin et al. (1980) showed that dietary supplementation of bile
salts improves emulsification, micelle formation and, consequently, fat digestibility. Also,
results of Zhang et al. (2011) demonstrated that a supplementation of lysophosphatidyl-
choline can improve fat digestion of broilers during the first 21 d of experiment. On the
other hand, some researchers reported that the addition of emulsifiers had no positive
effects on birds’ growth performance (Soares and Lopez-Bote 2002; Azman et al. 2004).
Glyceryl polyethylene glycol ricinoleate (GPR) is an emulsifying agent that may also
be used to enhance dietary fat availability. However, there is lack of information on the
use of GPR supplements in non-ruminants. Udomprasert and Rukkwamsuk (2006) found
that GPR may improve average daily weight gain and feed conversion ratio (FCR) of
weanling pigs. According to Amitava et al. (2010), a dietary addition of GPR at 1% may
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improve chickens’ live weight by up to 5% and, in this way, improves feed conversion.
The aforementioned authors showed that the GPR effect on fat utilisation was evidenced
from improvements in apparent total tract digestibility (ATTD) of fat and overall fat
metabolisability. In these trials conducted with piglets and chickens, the authors used
vegetable fats to increase the dietary energy concentration. According to Tancharoenrat
et al. (2013), in comparison with vegetable fats animal fats fed to broiler chickens are
characterised by poorer digestibility and a lower level of apparent metabolisable energy
(AMEN). The primary reason for inconsistency in results is therefore attributable to the
tested fat sources in which vegetable fats were more easily digestible than animal fats.
Moreover, the differences may be attributed to the interaction between the exogenous
emulsifier and the fat source.
According to the authors’ knowledge, in the literature no data are available on the
effects of GPR in broiler diets containing a blend of rapeseed oil and lard as fat source.
Therefore, the objectives of this trial were to determine the effects of GPR on growth
performance and nutrient digestibility in broiler chickens when they received high-fat
diets or diets with a reduced AMEN level achieved by a deteriorated fat content.

2. Materials and methods

2.1. Birds and experimental design
All animal procedures were conducted in accordance with the guidelines of the Polish
Council of Animal Care. The protocol for this study was approved by the Local Animal
Care Committee of the Poznan University of Life Sciences (permission number: 33/2013).
The experiment was conducted with 384 male broiler chickens (Ross 308) (initial
body weight 40 ± 2 g). The 12 replicate pens of 8 birds each were randomly assigned to 4
dietary treatments. All diets were based on wheat–maize–soybean meal with rapeseed oil
and lard (blend of 50:50) as fat sources (Table 1). Independent of allocation to an
experimental group, broilers received a starter feed until d 14 of experiment, which
contains less AMEN and more CP than the feed offered during the grower period from
d 14 to 35 of experiment (Table 1).
The AMEN contents of Diets SE met or exceeded recommendations for broiler
chickens (Aviagen 2009) (12.7 and 13.0 MJ/kg for starter and grower diets, respectively).
Diets LE were formulated to have a decreased AMEN content (0.41 and 0.42 MJ/kg less
for starter and grower diets, respectively), which was achieved by a reduction in the
rapeseed oil and lard contents (Table 1). The contents of other nutrients in Diets LE were
nearly similar to the respective Diets SE. Furthermore, in both fattening periods, Diets SE
 Archives of Animal Nutrition 287

Table 1. Composition and nutrient contents of the experimental diets (as feed basis).

Starter period (d 0–14) Grower period (d 14–35)

Diet SE* Diet LE§ Diet SE Diet LE

Ingredients [%]
Wheat 36.24 38.40 40.90 45.40
Soybean meal 31.86 31.36 23.86 23.15
Maize 20.00 20.00 21.00 21.00
Rapeseed meal 3.00 3.00 5.00 5.00
Rapeseed oil 2.74 1.91 3.08 2.18
Lard 2.74 1.91 3.08 2.18
Monocalciumphosphate 0.683 0.679 0.595 0.592
Limestone 0.336 0.337 0.274 0.258
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NaCl 0.15 0.15 0.154 0.154

NaHCO3 0.25 0.25 0.20 0.20
Lysine-HCl 0.283 0.291 0.260 0.272
Methionine 0.289 0.286 0.211 0.206
Threonine 0.104 0.105 0.064 0.076
Premix# 1.00 1.00 1.00 1.00
Econase XT† 0.01 0.01 0.01 0.01
Quantum Blue 5G‡ 0.01 0.01 0.01 0.01
TiO2 0.30 0.30 0.30 0.30
AMEN [MJ/kg] (calculated) 12.68 12.27 12.98 12.56
Analysed content (n = 8)
Dry matter [%] 88.5 88.5 88.1 88.7
Crude protein [%] 23.5 23.6 19.8 19.9
Ether extract [%] 7.16 5.65 7.99 6.23
Gross energy [MJ/kg] 18.1 17.6 18.1 17.7
Calcium [%] 0.90 0.89 0.83 0.84
Phosphorus [%] 0.57 0.56 0.54 0.55
TiO2 [%] 0.30 0.30 0.29 0.31
Neutral detergent fibre [%] – – 12.5 12.4
Notes: *SE, Energy supply according to standard recommendations; §LE, Low-energy supply (AMEN reduced);
#
Provides per kg diet: 11,250 IU vitamin A, 2500 IU cholecalciferol; 80 mg vitamin E, 2.50 mg menadione,
0.02 mg vitamin B12, 1.17 mg folic acid, 379 mg choline, 12.5 mg D-pantothenic acid, 7.0 mg riboflavin,
41.67 mg niacin, 2.17 mg thiamin, 0.18 mg D-biotin, 4.0 mg pyridoxine, 0.09 mg ethoxyquin, 73 mg Mn, 55 mg
Zn, 45 mg Fe, 20 mg Cu, 0.62 mg I, 0.3 mg Se, 60 mg salinomycin; †Enzyme premix, provides 16,000 BXU of
endo-1,4-beta-xylanase per kg diet; ‡Enzyme premix, provides 500 FTU of 6-phytase per kg diet.

and LE were offered either without supplementation of an emulsifier or supplemented

with commercially produced GPR (E 484, Bredol® 683; AkzoNobel SC AB,
Stenungsund, Sweden) at concentrations of 0.04%. When added to the experimental
diets, GPR was premixed with water. This procedure was followed by transferring the
screened and premixed portions to a stainless steel horizontal feed mixer (100 or 300
MPW, Zuptor sp. z o.o., Gostyń, Poland; mixing time was 4 min, mixing band: 27.4 rev/
min) for mixing of the completed diet. All ingredients except minerals, vitamins, amino
acids and fat were ground in a Skiold Disk mill (SK2500, Skiold A/S, Sæby, Denmark)
with disc distance set at 1.8 mm. All diets were provided ad libitum in mesh form from d
1 to 35 and were supplemented with titanium dioxide (TiO2) as marker to calculate the
digestibility. To provide a commercial formulation, exogenous enzymes were used. All
diets were supplemented with 0.01% of Econase XT 25P (AB-Vista, Marlborough, UK,
 288 S. Andrzej Kaczmarek et al.

providing 16,000 BXU of endo-1,4-beta-xylanase per kg diet) and 0.01% of Quantum

Blue 5G (AB-Vista, Marlborough, UK, providing 500 FTU of 6-phytase per kg diet).
For estimation of nutrient digestibility, collection trays were installed in floor pens on
d 14 and 35 to allow excreta collection. For this purpose, all birds were removed from
pens and wire floor trays were placed above the collection tray. After approximately 3 h,
five excreta samples per treatment, free from contamination like feed or feathers, were
collected. Five excreta samples per treatment were collected and one excreta sample
represented one pen (eight birds).

2.2. Chemical analysis

Feed samples were analysed in duplicate for crude protein, ether extract (EE), crude fibre,
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Ca and total P using AOAC (2005) methods: 976.05, 920.39, 2002.04 and 965.17,
respectively. For all chemical analyses, samples were ground to pass a 1 mm sieve.
Prior to analysis, excreta samples were homogenised by a stomacher homogeniser
(Interscience, Saint-Nom-la-Bretèche, France), then freeze-dried (Christ 1825
Medizinischer Apparatebau 326; Martin Christ GmbH, Osterode, Germany) and ground
(1 mm screen) excreta samples were analysed for nitrogen, EE and neutral detergent fibre
(NDF) using AOAC (2005) methods. TiO2 was determined according to the method of
Short et al. (1996); the samples were prepared according to the procedure proposed by
Myers et al. (2004). Gross energy was determined using an adiabatic bomb calorimeter
(KL 12Mn, Precyzja-Bit PPHU, Poland) standardised with benzoic acid.
Fatty acid profile of diets was determined according to Cieślak et al. (2009) using a
gas chromatograph (GC Bruker 456-GC, USA). A Chrompack CP7420 column (100 m,
0.25 mm, 0.25 µm film thickness, Agilent HP) and a flame ionisation detector were used.
Hydrogen was used as carrier gas at a constant flow of 40 psi. The oven temperature was
programmed as follows: initially 120°C for 7 min, then increasing at 7°C/min to 140°C
holding for 10 min and then increasing at 4°C/min to 240°C. A sample volume of 2 µl
was injected into the column. The qualitative and quantitative identification of fatty acid
peaks was made using a method based on external standards comparing to Supelco 37
Component FAME Mix (Sigma Aldrich, Bellefonte, PA, USA) using Galaxie Work
Station 10.1 (Varian, Walnut Creek, CA, USA).

2.3. Calculations and statistical analysis

ATTD and AMEN contents of the diets were calculated relative to the ratio of TiO2 to the
content of the nutrient in question in the feed or excreta. Digestibility of various dietary
components (using fat calculation as an example) and AMEN content of experimental
diets were calculated according to following equations:

Digestibility½%¼f1  ½ðTiO2diet =TiO2excreta ÞðFatexcreta =Fatdiet Þg100%;

where the contents of TiO2 and fat in diets and excreta are given as %.
AMEN [MJ/kg] =

GEdiet  ½GEexcreta ðTiO2diet =TiO2excreta Þ  0:0344fNdiet  ½Nexcreta ðTiO2diet =TiO2excreta Þg;

 Archives of Animal Nutrition 289

where GE is the gross energy [MJ/kg], N is nitrogen [%] and TiO2 [%] is the dietary
marker. The AME value was corrected to zero nitrogen balance using 34.4 MJ/kg N
retained (Hill and Anderson 1958).
Two-way analysis of variance was performed using the R environment (R Development
Core Team 2014) and using the “agricolae” package (De Mendiburu 2014) according to the
following general model:

Yij ¼ μ þ αi þ βj þ ðαβÞij þ εij ;

where Yij is the measured dependent variable, µ is overall mean, αi is the effect of diet (SE
or LE), βj is the effect of emulsifier (αβ) is the interaction between diet and emulsifier and
εij is the random error. Means were compared pairwise using Duncan’s multiple range test.
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In the event of absence of interactions significances, main effects were discussed. All data
are presented as means with pooled standard error of the mean (SEM).

3. Results
The analysed chemical and fatty acid composition of the experimental diets is presented in
Tables 1 and 2. All analysed nutrients were recovered at concentrations similar to
formulated values, confirming the accuracy of formulation and mixing. All diets were

Table 2. Fatty acid composition [% of total fatty acids] and ratio of unsaturated to saturated fatty
acids in the experimental diets.

Starter period (d 0–14) Grower period (d 14–35)

Diet SE* Diet LE§ Diet SE Diet LE

C6:0 (Caproic acid) 0.11# 0.1 0.12 0.12

C8:0 (Caprylic acid) 0.09 0.08 0.12 0.14
C10:0 (Capric acid) 0.08 0.07 0.09 0.07
C12:0 (Myristic acid) 0.10 0.11 0.15 0.10
C14:0 (Myristic acid) 0.06 0.04 0.05 0.07
C16:0 (Palmitic acid) 17.4 17.4 18.0 17.9
C18:0 (Stearic acid) 7.30 6.21 7.31 6.24
C21:0 (Heneicosanoic acid) 0.65 0.40 0.67 0.41
C23:0 (Tricosanoic acid) 0.19 0.15 0.20 0.15
C24:0 (Lignoceric acid) 0.17 0.15 0.18 0.15
Sum of saturated fatty acids 26.2 24.7 26.9 25.1
C16:1 (Palmitoleic acid) 0.60 0.23 0.59 0.21
C18:1 (Oleic acid) 40.1 37.7 40.1 37.5
C18:1 (Vaccenic acid) 2.55 2.5 2.54 2.46
C18:2 (Linoleic acid) 24.5 29.0 23.8 28.7
C18:3 (Linolenic acid) 5.00 5.10 4.70 5.00
C20:2 (Eicosadienoic acid) 0.23 0.14 0.17 0.13
C22:1 (Erucic acid) 0.22 0.2 0.24 0.21
C20:3 (Dihomo-γ-linolenic acid) 0.10 0.31 0.13 0.40
C20:4 (Arachidonic acid) 0.20 0.11 0.23 0.15
Sum of unsaturated fatty acid 73.9 75.3 73.1 74.9
Ratio unsaturated to saturated fatty acids 2.82 3.05 2.72 2.98

Notes: *SE, Energy supply according to standard recommendations; §LE, Low-energy supply (AMEN reduced);
#
n = 2.
 290 S. Andrzej Kaczmarek et al.

characterised by a high EE content. The determined fatty acid profile showed high content
of unsaturated fatty acids (>70%). The dominating unsaturated fatty acids were oleic acid
(38–40%) and linoleic acid (24–29%). The dominating saturated fatty acid was palmitic
acid (17–18%). Differences in fatty acid composition between Diets SE and LE were
negligible.
The overall mortality of approximately 4% (0–35 d of experiment) was not related to
dietary treatment at any phase measured (data not shown).
During the first 2 weeks of experiment, there was no effect of GPR inclusion on the
performance of birds (Table 3) and broilers were characterised by similar body weight
gain (BWG), feed intake (FI) and FCR. From d 14 to 35, the decreased AMEN level in the
not supplemented Diet LE caused a reduced BWG (~1.5%, p < 0.05). GPR supplementa-
tion improved BWG, particularly when Diet LE was fed, resulting in an interaction
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between GPR inclusion and AMEN level (p = 0.016). The experimental treatments did
not affected FI at any time measured (Table 3). During the grower period, birds fed diets
with GPR were characterised by a lower FCR (p = 0.002), and during the whole
experimental period, this effect was still significant for Diet LE. No significant interac-
tions between treatments on FCR were observed.
In the starter period, the ATTD of EE was increased by GPR inclusion (p < 0.05), but
not by AMEN level of diet (Table 4). There was a tendency of an improved AMEN level
after GPR inclusion (p < 0.1). In Diet LE, the determined AMEN level was by 0.40 MJ/kg
lower than in Diet SE. During d 14–35 of experiment, the GPR supplementation improved
the ATTD of GE only in Diet LE (p < 0.05), resulting in a significant interaction between
GPR inclusion and AMEN level (Table 5). Furthermore, the GPR supplementation
resulted in an increased ATTD of NDF (p < 0.05), independent of AMEN level of diet.
However, this improvement of ATTD by GPR addition was greater for Diet LE, resulting
in a significant interaction between GPR inclusion and AMEN level. Multiple compar-
isons showed that the ATTD of EE was significantly improved after GPR addition, but
there were no interactions of treatment effects revealed. Finally, a significant improvement
of the AMEN level after GPR addition was only shown for Diet LE and as main effect a
significantly higher AMEN level of Diets SE vs. Diets LE were observed (difference:
0.46 MJ/kg, p < 0.05). This resulted in a significant interaction between GPR inclusion
and AMEN level.
Considering the entire experimental period, the use of GPR improved the broiler
BWG when the energy-reduced Diets LE were fed and the ATTD of EE at the end of the
grower period (d 35). At d 14 of experiment, GPR inclusion tended to improve the dietary
AMEN contents irrespective of the applied diet, but at d 35 of experiment this improve-
ment was noticeable only in birds fed Diet LE.

4. Discussion
Given the relatively high-energy requirements of recent genotypes of broiler chickens, it is
common practice to increase their dietary energy contents by high inclusion rates of fat.
Tancharoenrat et al. (2013, 2014) demonstrated that the utilisable AME contents and
digestibility of fat were influenced by the age of broilers. They concluded that the lower
AME and fat digestibility determined during week 1 of life confirm that the physiological
ability to digest and absorb fats is immature at hatch, but this ability develops rapidly with
progressing age. Furthermore, the digestibility of dietary fat depends on the chemical
nature of its constituent fatty acids. Fats rich in unsaturated fatty acids are better digested
and absorbed than saturated fats (Dänicke 2001).
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Table 3. Performance of birds fed Diets SE* or LE#, supplemented or not with glyceryl polyethylene glycol ricinoleate (GPR) (n = 10).

Starter period (d 0–14) Grower period (d 14–35) Whole experimental period (d 0–35)
♦ ‡
BWG Feed intake FCR BWG Feed intake FCR BWG Feed intake FCR
GPR inclusion† AMEN level [kg/bird] [kg/bird] [kg:kg] [kg/bird] [kg/bird] [kg:kg] [kg/bird] [kg/bird] [kg:kg]

− SE 0.412a 0.515a 1.25a 1.627a 2.591a 1.59a 2.04a 3.105a 1.52ab

− LE 0.386a 0.493a 1.28a 1.566b 2.554a 1.63a 1.95b 3.047a 1.56a
+ SE 0.388a 0.480a 1.24a 1.656a 2.567a 1.55b 2.04a 3.047a 1.49b
+ LE 0.390a 0.500a 1.28a 1.668a 2.592a 1.55b 2.06a 3.092a 1.50b
SEM§ 0.006 0.007 0.013 0.009 0.012 0.010 0.011 0.017 0.007
Main effects
− 0.399 0.504 1.27 1.598 2.573 1.61 1.997 3.078 1.54
+ 0.388 0.490 1.26 1.662 2.579 1.55 2.050 3.069 1.50
SE 0.400 0.498 1.25 1.641 2.579 1.57 2.041 3.077 1.51
LE 0.388 0.497 1.28 1.617 2.573 1.59 2.005 3.069 1.53
p-Value
GPR inclusion 0.384 0.346 0.866 <0.001 0.804 0.002 0.005 0.805 0.001
AMEN level 0.329 0.934 0.163 0.098 0.789 0.248 0.044 0.821 0.067
GPR × AMEN 0.256 0.163 0.798 0.016 0.200 0.318 0.007 0.129 0.339
Notes: *SE, Energy supply according to standard recommendations; #LE, Low-energy supply (AMEN reduced); †Supplementation with GPR is indicated by +, no supplementation by
–; ♦BWG, body weight gain; ‡FCR, feed conversion ratio; §SEM, standard error of the mean.
ab
Means in the same column with different superscripts are significantly different at p < 0.05.
Archives of Animal Nutrition
291
 292 S. Andrzej Kaczmarek et al.

Table 4. Influence of dietary energy level (SE* or LE#) and inclusion of glyceryl polyethylene
glycol ricinoleate (GPR) on dietary AMEN level and apparent total tract digestibility (ATTD) of
ether extract on d 14.

GPR inclusion† AMEN level ATTD of ether extract AMEN [MJ/kg]

– SE 0.777a 12.62a
– LE 0.771a 12.20b
+ SE 0.822a 12.82a
+ LE 0.814a 12.44ab
SEM‡ 0.059 0.494
Main effects
– 0.774 12.41
+ 0.818 12.63
SE 0.800 12.72
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LE 0.793 12.32
p-Value
GPR inclusion 0.011 0.092
AMEN level 0.667 0.003
GPR × AMEN 0.958 0.878

Notes: *SE, Energy supply according to standard recommendations; #LE, Low-energy supply (AMEN reduced by
0.419 MJ/kg); †Supplementation with GPR is indicated by +, no supplementation by –; ‡SEM, Standard error of
the mean;
ab
Means in the same column with different superscripts are significantly different at p < 0.05.

Table 5. Influence of dietary energy level (Diets SE* or LE#) and inclusion of glyceryl poly-
ethylene glycol ricinoleate (GPR) on AMEN level of diet, apparent total tract digestibility (ATTD) of
gross energy (GE), neutral detergent fibre (NDF) and ether extract (EE) on d 35.

ATTD

GPR inclusion† AMEN level GE NDF EE AMEN [MJ/kg]

− SE 0.704 ab
0.066 bc
0.814 b
12.89a
− LE 0.668c 0.028c 0.796c 11.97b
+ SE 0.695b 0.124ab 0.845a 12.61a
+ LE 0.722a 0.182a 0.846a 12.62a
SEM‡ 0.044 0.138 0.041 0.077
Main effects
− 0.686 0.047 0.805 12.43
+ 0.701 0.153 0.846 12.61
SE 0.700 0.095 0.829 12.75
LE 0.695 0.105 0.821 12.29
p-Value
GPR inclusion 0.002 <0.001 <0.001 0.151
AMEN level 0.504 0.667 0.128 <0.001
GPR × AMEN <0.001 0.036 0.750 <0.001

Notes: *SE, Energy supply according to standard recommendations; #LE, Low-energy supply (AMEN reduced by
0.419 MJ/kg); †Supplementation with GPR is indicated by +, no supplementation by –; ‡SEM, Standard error of the
mean.
abc
Means in the same column with different superscripts are significantly different at p < 0.05.

Surprisingly, in the present study, the addition of GPR had no effect on broiler
performance from d 0 to 14 of experiment, but affected the performance during the
grower period and subsequently in the whole experiment. Serafin and Nesheim (1970)
and Noy and Sklan (1995) attributed the poor digestibility of dietary fat to the limited
 Archives of Animal Nutrition 293

secretion of lipase and/or bile salts by chicks. On the other hand, Meng et al. (2004)
speculated that the deficient secretion of lipase and bile salts in young birds, when
calculated per gram of FI, is less dramatic. This indicates that the lipase secretion of
young birds may not be as inadequate as expected when their FI is considered. This could
explain the lack of response of birds after GPR inclusion (d 0–14). However, in this study,
the GPR inclusion improved the ATTD of EE and tended to enhance dietary AMEN on d
14. Additionally, fat content in practical broiler starter diets is low. In the starter period of
this study, Diet LE contained about 5.7% EE (Table 1) and the average daily feed intake
was approximately 35 g/bird (Table 3). This means that birds consumed approximately
2 g fat per day. Based on these data and digestibility results (Table 4), it can be concluded
that after GPR inclusion the improvement in EE utilisation was only about 0.09 g digested
EE per bird per day and in the whole starter period only 1.2 g digested EE. The above-
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mentioned data and the inherent bird variability as well as measurement accuracy may
explain why birds failed to respond to GPR inclusion during the starter period. Al-
Marzooqi and Leeson (1999) reported that using an emulsifier alone or in combination
with lipase had no effect on performance results irrespective of age of birds. In contrast to
Al-Marzooqi and Leeson (1999), who used diets based on maize, in our trial wheat–maize
diets were used. It is well known that maize-based diets are less viscous than wheat-based
diets. The effect of viscosity on fat emulsification and digestion is well accepted in
literature (Choct and Annison 1992; Dänicke, Jeroch, et al. 1999; Dänicke, Simon, et
al. 1999; Meng et al. 2004). It could be speculated that discrepancies between the
previously performed trial (Al-Marzooqi and Leeson 1999) and the presented results
may be attributed to differences in diet composition and its viscosity. On the other
hand, Amitava et al. (2010) found that an exogenous emulsifier improved the performance
of birds during the starter and grower periods, but Zhang et al. (2011) found such a
positive effect only during the starter period. In both investigations (Amitava et al. 2010;
Zhang et al. 2011), maize-based diets were used. Other researchers found that exogenous
emulsifiers may improve the performance of weanling pigs but some inconsistency in
results was also observed (Øverland and Sundstøl 1995; Soares and Lopez-Bote 2002;
Udomprasert and Rukkwamsuk 2006).
Zhang et al. (2011) and Amitava et al. (2010) concluded that the improved perfor-
mance after addition of an emulsifier may be related to the observed improvement of
digestibility of crude fat or of selected fatty acids. Throughout this experiment, the
emulsifier improved the digestibility of EE at d 14 and 35. Consequently, as ATTD of
EE increased after GPR inclusion, a tendency of a higher AMEN level for Diets SE and
LE was observed on d 14 (Table 4). After GPR inclusion, the ATTD of GE was improved
for Diet LE only. These results appear to contradict those obtained by Al-Marzooqi and
Leeson (1999), who reported that the presence of a supplemental emulsifier in the diets of
broiler chickens was not proved to be an effective method to improve fat digestibility and
increase the AME level of diet. On the other hand, our findings agree with the report of
Zhang et al. (2011), who showed that emulsifier addition improved the AME level of
diets. However, the reason behind the failure of the emulsifier to improve the dietary
AME level on d 35 is unknown. It could be speculated that older birds were characterised
by larger capacity of digestive tract, which may mask the positive effects of the emulsifier.
ATTD of EE and GE obtained in this study clearly showed that the GPR-supplemented
diets were utilised better. The determined EE digestibility in this trial was high and
increased with age of birds. These findings are in general agreement with those of
Tancharoenrat et al. (2013, 2014), who reported that the ATTD of EE (from soybean
oil) increased with age of birds and was almost 60% at first week and 95% at fifth week of
 294 S. Andrzej Kaczmarek et al.

bird’s life. In this study, the high fat digestibility in rapeseed oil was expected because it
contains high concentrations of unsaturated fatty acids (Table 2) which are better digested
and absorbed by poultry than saturated ones (Tancharoenrat et al. 2014). Crude rapeseed
oil is primarily composed of long-chain unsaturated fatty acids: oleic, linoleic and
linolenic. Garrett and Young (1975) found that long-chain unsaturated fatty acids could
be easily absorbed even in the absence of bile salts. Additionally, in this study, a blend of
50:50 lard and rapeseed oil was used. It is known that the utilisation of saturated fatty
acids is improved in the presence of high concentrations of unsaturated fatty acids (Young
and Garrett 1963; Tancharoenrat et al. 2013, 2014). High concentrations of unsaturated
fatty acids can act as emulsifiers (Tancharoenrat et al. 2013, 2014) to enhance the
emulsification of saturated fatty acids. The analysed dietary fatty acid profile (Table 2)
showed that mainly unsaturated fatty acids prevailed. This may explain the observed high
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ATTD of EE irrespective of the addition of an emulsifier.

In our experiment, the expected low ATTD of NDF was higher when diets were
supplemented with GPR, resulting in a GPR inclusion × AMEN interaction. Nevertheless,
to the authors’ knowledge, in the literature no data are available about the effect of fat
level or addition of emulsifiers on fibre digestibility in birds. However, addition of fats to
ruminant diets reduced fibre digestibility due to the following reasons: physical coating of
the fibre with fat, modification of rumen microbial population and reduction in cation
availability (Palmquist and Jenkins 1980). Lee et al. (2004) found that surfactants might
improve in vitro and in situ rumen fermentation. The aforementioned authors observed
higher ruminal microbial population and hydrolytic enzyme activities in rumen fluids. The
use of GPR in this study improved EE digestibility, so it could be speculated that the
observed changes in total tract NDF digestibility may be related to enhanced accessibility
of microbes to fibre and higher activity of bacteria in lower parts of the gastrointestinal
tract. On the other hand, degradation of fibre-fractions determined using marker method
has limitations. According to De Vries et al. (2014), when degradation of fibre-fractions is
the matter of interest, separation of marker and specific digesta fractions occurs, the
marker method has limitations. According to the aforementioned authors, digestibility
values determined using the marker method are somewhat lower than those obtained by
total collection, particularly in fibre-rich diets. The above data show that care should be
taken when the marker method is used to estimate fibre-fractions digestibility, and the
result obtained for NDF digestibility in this study should be confirmed by further trials.
In conclusion, the findings of the current study suggest that the positive effects of
GPR on blend of animal fat/rapeseed oil digestion are substantial even in a practical
maize–wheat-based broiler diet with decreased AMEN level. Consequently, the
applied emulsifier improved BWG of chickens fed Diets LE and FCR in both Diets LE
and SE.

Disclosure statement
No potential conflict of interest was reported by the authors.

ORCID
Sebastian Andrzej Kaczmarek http://orcid.org/0000-0003-3638-4192
 Archives of Animal Nutrition 295

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