Effect of Extrusion on Nutrients Digestibility, Metabolizable Energy and Nutritional Value of Yellow Lupine Seeds for Broiler Chickens

Abstract

The aim of investigations was the estimation of nutritional value of actual cultivated yellow lupine cultivars in raw and extruded form, and their usefulness for broiler chickens. Two experiments were conducted with male Ross 308 chickens. In a digestibility trial 60 fifteen-day old birds were randomly assigned to three dietary treatments (20 replications in each). Birds were kept in individual pens. Digestibility was calculated using the difference method. From day 16 to 21, diets contained lupine meal in raw or extruded form and the basal diet in the ratio 20:80. Subsequently, the total digestibility of dry matter and crude fat, also apparent nitrogen retention and AME
In the second experiment the one day old birds were randomly assigned to ten dietary treatments (10 replications in each) and were fed diets with increasing amounts of raw or extruded lupine from 10 to 30%. The chickens in control treatments were fed a diet without lupine.
Extruded yellow lupine meal was characterized by lower phytic P content in comparison to raw yellow lupine meal. The content of remaining ingredients and antinutritional substances was similar. Yellow lupine seeds post extrusion were characterized by better total crude fat digestibility, nitrogen retention and AME
By applying diets in the broilers consisting of 10 to 30% of lupine seeds post extrusion (experiment II), improved apparent fat digestibility, apparent nitrogen retention and AME

Full text

Ann. Anim. Sci., Vol. 16, No. 4 (2016) 1059–1072 DOI: 10.1515/aoas-2016-0025
EFFECT OF EXTRUSION ON NUTRIENTS DIGESTIBILITY,
METABOLIZABLE ENERGY AND NUTRITIONAL VALUE OF YELLOW
LUPINE SEEDS FOR BROILER CHICKENS* *
Andrzej Rutkowski1, Sebastian A. Kaczmarek1, Marcin Hejdysz1♦, Dorota
Jamroz
2
1Department of Animal Nutrition and Feed
Management,
Poznan University of Life Sciences,
Wołyńska 33, 60-637 Poznań, Poland
2Department of Animal Nutrition and Feed Quality, Wrocław University of Environmental
and Life Sciences, Chełmońskiego 38 C, 51-630 Wrocław, Poland
♦Corresponding author: marhej@up.poznan.pl
Abstract
The aim of investigations was the estimation of nutritional value of currently cultivated yellow
lupine cultivars in raw and extruded form, and their usefulness for broiler chickens. Two experi-
ments were conducted with male Ross 308 chickens. In a digestibility trial 60 fteen-day-old birds
were randomly assigned to three dietary treatments (20 replications in each). Birds were kept in
individual pens. Digestibility was calculated using the difference method. From day 16 to 21, diets
contained lupine meal in raw or extruded form and the basal diet in the ratio 20:80. Subsequently,
the total digestibility of dry matter and crude fat, also apparent nitrogen retention and AMEN
value of lupine seeds were determined on chickens fed different forms of lupine. The ileal digest-
ibility of crude protein and amino acids of lupine seeds was also analyzed. In the second experi-
ment the one-day-old birds were randomly assigned to ten dietary treatments (10 replications in
each) and were fed diets with increasing amounts of raw or extruded lupine from 10 to 30%. The
chickens in control treatments were fed a diet without lupine. Extruded yellow lupine meal was
characterized by lower phytic P content in comparison to raw yellow lupine meal. The content of
remaining ingredients and antinutritional substances was similar. Yellow lupine seeds post extru-
sion were characterized by better total crude fat digestibility, nitrogen retention and AMEN values,
compared to raw seeds. Higher ileal digestibility was conrmed in numerous amino acids, except
lysine, threonine and valine (P≤0.05). By feeding the broilers with diets consisting of 10 to 30% of
lupine seeds post extrusion (experiment II), improved apparent fat digestibility, apparent nitrogen
retention and AMEN values were achieved in young chickens (P<0.01). Using 10 and 20% of lupine
in the diets showed signicant positive effects of extrusion on body weight gains, feed intake and
feed conversion rate. The performance indices of chickens were drastically decreased by use of
25% ratio of both raw and extruded yellow lupine in the diet. This effect was heightened by a 30%
share in feed mixtures.
Key words: broiler chickens, lupine, extrusion, digestibility
*The study was realized within the research project “Improvement of native protein, feeds, their
production, trade turnover and utilization in animal feed” nanced by the Ministry of Agriculture and
Rural Development of Poland NO 505.037.07.
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A. Rutkowski et al.
1060
Legume seeds are an important protein source for humans and animals in many
countries. Yellow lupine protein is rich in lysine, but it is decient in sulfur-contain-
ing amino acids and leucine (Sujak et al., 2006). Lupine seeds are rich in crude fat
~5–9%, but also the content of dietary ber is relatively high (Gdala, 1998; Zalewski
et al., 2001; Zduńczyk et al., 2014; Krawczyk et al., 2015; Kaczmarek et al., 2016).
Legume seeds, including lupines, are also rich in microelements, e.g., Mn, Zn, Cu
(Wasilewko and Buraczewska, 1999) and antioxidants – selenium and tocopherol
(Erbaş et al., 2005).
In the past, the use of yellow lupine (Lupinus luteus) as a valuable source of pro-
tein for poultry was limited due to high alkaloid and non-starch polysaccharide (NSP)
contents that negatively affect growth, feed intake and nutrient utilization (Kluge et
al., 2002; Olkowski et al., 2005). Current yellow lupine cultivars are characterized
by low concentrations of alkaloids, but they still contain other anti-nutritional fac-
tors, which can reduce nutrient digestibility. The main lupine anti-nutritional factors
are: α-galactosides (Gdala, 1998; Zalewski et al., 2001), proteases and α-amylase in-
hibitors (Erbaş et al, 2005; Guillamon et al., 2008), saponins, quinolizidine alkaloids
(Wasilewko and Buraczewska, 1999; Mańczak et al., 2007), glycosides, phytates (Jul
et al., 2003). According to some researchers quinolizidine and indole alkaloids and
rafnose content depends on lupine species and variety (Smulikowska et al., 1999;
Wasilewko and Buraczewska, 1999; Kaczmarek et al., 2014; Rutkowski et al., 2015).
According to available literature data (Orda et al., 2006; Rutkowski et al., 2015), it
has been shown that yellow lupine can be a benecial component of poultry diets.
The nutritive value of seeds can be efciently improved by extrusion (Leontowicz
et al., 2001; Kiczorowska and Lipiec, 2002 a, b). The extrusion process reduces the
activity of thermolabile harmful substances, decreases the negative action of oligo-
saccharides and protease inhibitors, and can inactivate pathogens (Guillamon et al.,
2008; Nalle, 2009). Additionally, according to Alonso et al. (2000) extrusion de-
creases phytic P content in seeds because some molecules of inositol hexaphosphate
are hydrolyzed to penta-, tetra- and triphosphates. However, the effect of extrusion
on the nutritive value of yellow lupine is not well understood. Moreover, there is no
comparative information on the inuence of extrusion on the nutritive value of yel-
low lupine and broiler chickens performance.
The objective of this study was to determine AMEN and digestibility of selected
nutrients of raw and extruded yellow lupine seeds and compare the effects of the use
of different amounts of raw or extruded yellow lupine seeds in place of soya bean
meal in broiler diets.
Material and methods
All procedures used in the experiment were accepted by the Local Ethics
Commission with respect to animal experimentation.
Lupine seed and processing
Raw or extruded yellow lupine (Lupinus luteus) cv. Mister seeds harvested in
2014 were used. Seeds were extruded with KNZ 2 extruder (Russia) (500 kg/h), at
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Raw and extruded lupine seeds in broiler diets 1061
temperature 135±10°C, pressure 30 kg/m2, cylinder speed 1000 rotations per minute.
Seeds were analyzed for basal nutrients, amino acids and anti-nutritive substances
(Table 1).
Table 1. Chemical composition of raw and extruded yellow lupine seeds var. Mister
Item Lupine seeds
raw extruded
Dry matter (%) 89.01 89.35
Crude ash (% DM) 4.15 4.14
Crude protein (% DM) 38.98 39.12
Crude ber (% DM) 19.23 19.18
ADF (% DM) 24.24 23.96
NDF (% DM) 28.24 28.41
Crude fat (% DM) 5.26 5.42
Gross Energy (MJ/kg DM) 20.49 20.55
WEV (cP) 1.09 1.17
Amino acid (g/16 g N)
Asp 8.81 8.64
Thr 3.17 3.21
Ser 4.24 4.34
Glu 24.46 24.05
Pro 6.08 6.16
Gly 3.47 3.52
Ala 2.83 2.85
Val 3.17 3.26
Ile 3.20 3.28
Leu 6.50 6.52
Tyr 3.24 3.18
Phe 4.24 4.17
His 3.32 3.22
Lys 4.76 4.81
Arg 10.12 10.30
Anti-nutrients in DM
Total alkaloids (mg/kg) 27.0 28.0
Lupinine, in total alkaloids (%) 63.29 64.58
Sparteine, in total alkaloids (%) 33.6 34.1
Ammodendrine, in total alkaloids (%) 3.12 3.18
Oligosaccharides (%) 8.57 8.61
Rafnose (%) 1.10 1.14
Stachyose (%) 4.94 4.97
Verbascose (%) 2.53 2.50
P phyt. (%) 0.70 0.59
P phyt./P total 75 63
WEV – water extract viscosity.
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A. Rutkowski et al.
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Bird management and sample collection
Experiment 1
The experiment was conducted on sixty 16-day-old Ross 308 male chickens,
reared in the rst days of life in pens and fed basal diets (BD) (Table 2). At day 16
of age the birds were randomly allocated to individual pens and assigned to three
dietary treatments, each with 20 replications. Chickens in control treatment were fed
BD, in treatment 2 with 80% BD + 20% raw lupine, and in treatment 3 with 80%
BD + 20% of extruded lupine. For the determination of total or ileal digestibility as
well as AMEN, 3 g/kg of titanium dioxide was included into diets as a non-absorbable
marker. The difference method for the digestibility calculation was used (Hejdysz et
al., 2015). All diets were offered ad libitum in mash form.
Table 2. Composition of basal diet, % (Experiment 1)
Components g/kg–1
Maize 600.0
Soya bean meal 293.5
Soya oil 41.6
Fish meal 29.4
Monocalcium phosphate 10.3
Limestone 5.1
Premix110.0
NaCl 2.0
NaHCO30.1
DL-methionine 2.7
L-lysine 1.7
L-threonine 0.6
TiO23.0
Calculated
Metabolizable energy (MJ/kg) 12.5
Analyzed
Crude protein (g/kg–1) 212
1provides per kg diet: IU: vit. A – 11250, cholecalciferol – 2500; mg: vit. E – 80, menadione – 2.50, vit. B12 – 0.02,
folic acid – 1.17, choline – 379, D-pantothenic acid – 12.5, riboavin – 7.0, niacin – 41.67, thiamin – 2.17, D-biotin –
0.18, pyridoxine – 4.0, ethoxyquin – 0.09, Mn – 73, Zn – 55, Fe – 45, Cu – 20, I – 0.62, Se – 0.3, salinomycin – 60;
g: Ca – 3.8.
For purposes of acclimatization, during ve days, birds were fed experimental di-
ets. On days 19 and 20 of age, excreta were individually collected, daily, and immedi-
ately frozen for analyses (n=20/treatment). On day 21 of age, all chickens from each
group were sacriced by cervical dislocation and their ileum removed. Digesta were
ushed from the terminal ileum (15 cm, adjacent to the ileocecal junction) and pooled
(4 birds/sample) to provide sufcient material for chemical analyses (n=5).
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Raw and extruded lupine seeds in broiler diets 1063
Table 3. Composition of starter and grower diets, g/kg–1 (Experiment 2)
Component
Starter Grower
L30 L25 L20 L10 CL30 L25 L20 L10 C
Maize (9.4)1333.6 372.7 380.1 389.5 395.0 305.4 343.3 356.6 361.5 367.2
Yellow lupine (37.8)1300.0 250.0 200.0 100.0 - 300.0 250.0 200.0 100.0 -
Wheat (11.8)1100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0
Soya oil 75.0 66.0 66.0 60.0 56.0 100.0 92.0 87.0 85.0 81.0
Soya bean meal (41.7)1- 50.0 100.0 200.0 300.0 - 50.0 100.0 200.0 300.0
Rapeseed meal (34.9)150.0 40.0 40.0 40.0 40.0 50.0 40.0 40.0 40.0 40.0
DDGS (36)150.0 40.0 40.0 40.0 40.0 50.0 40.0 40.0 40.0 40.0
Pea (22.6)150.0 40.0 40.0 40.0 40.0 50.0 40.0 40.0 40.0 40.0
Limestone (<2mm) 6.2 6.2 - - - 6.2 6.2 - - -
Monocalcium phosphate 13.1 13.2 13.2 12.5 12.5 13.2 13.3 13.2 12.5 12.5
Premix210.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0
NaHCO33.1 3.2 3.2 2.5 2.2 3.6 3.7 3.7 3.2 2.5
NaCl 0.5 0.6 0.7 1.2 1.5 0.4 0.6 0.6 1.0 1.5
L-lysine HCL 4.1 3.8 3.4 2.3 1.2 3.5 3.3 2.8 1.7 0.6
DL-methionine 1.9 1.9 1.9 1.7 1.6 2.0 2.0 2.0 1.8 1.7
L-threonine 1.2 1.1 0.9 0.3 - 1.3 1.2 1.0 0.3 -
L-valine 1.3 1.3 0.6 - - 1.4 1.4 0.9 - -
TiO2- 3.0 3.0 3.0 3.0 3.0
Calculated
Metabolizable energy (MJ/kg) 12.2 12.2 12.3 12.1 12.1 12.9 12.9 13.0 12.9 12.9
Analyzed
Crude protein (g/kg) 220 221 220 223 220 210 211 209 2 11 210
1 % crude protein; 2provides per kg diet: IU: vit. A – 11250, cholecalciferol – 2500; mg: vit. E – 80, menadione – 2.50, vit. B12 – 0.02, folic acid – 1.17, choline – 379, D-
pantothenic acid – 12.5, riboavin – 7.0, niacin – 41.67, thiamin – 2.17, D-biotin – 0.18, pyridoxine – 4.0, ethoxyquin – 0.09, Mn – 73, Zn – 55, Fe – 45, Cu – 20, I – 0.62, Se – 0.3,
salinomycin – 60; g: C – 3.8.
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A. Rutkowski et al.
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Experiment 2
Eight hundred one-day-old male Ross 308 chickens of average initial weight
44.3 g were randomly assigned to ten treatments. Each treatment comprised ten rep-
lications of 8 birds each. Chickens were kept in pens (0.5 m2, each with 8 birds) on
straw litter. The environmental conditions were typical for broiler rearing; the light-
ing program in the rst 7 days was 24 h/day, and after that 18h light: 6h darkness.
Birds were reared in two identical rooms with controlled environmental conditions.
Composition of diets was shown in Table 3. Control diet was based on maize
and wheat, and contained soybean meal (SBM), rapeseed meal, distilled dried grain
with solubles (DDGS) and pea as protein sources. In experimental diets SBM was
replaced (w/w) by growing amounts of ground yellow lupine seeds, raw or extruded
(Table 3). Control diets (5 and 10) did not contain lupine seeds. Diets were isoen-
ergetic and isonitrogenous. The chickens were fed starter (1–14 days) and grower
diets (15–35 days) in mash form. Diets and water were accessible ad libitum. The
composition of diets was calculated based on chemical analyses of feed compounds
applied using simple linear optimization.
The body weight gains (BWG) of chickens and feed intake (FI) were measured
for each cage after day 14 and day 35 of experiment and mortality was registered as
it occurred.
In Experiment 2, to allow the digestibility to be determined, 3 g/kg titanium di-
oxide was included as a non-absorbable marker in diets fed during days 33 and 34
of growth experiment. The oor of each cage was covered with thick plastic foil and
excreta were collected twice a day. The samples were immediately homogenized and
frozen for chemical analyses (n=10) similar as in Experiment 1.
Chemical analyses
For chemical analyses, representative samples of seeds and diets were ground
to pass through a 0.5 mm sieve. Raw and extruded seeds, diets, ileum digesta and
excreta were analyzed in duplicate for dry matter (DM), nitrogen retention, ether
extract (EE), using methods AOAC (2007) no: 934.01, 976.05, 920.39, respectively.
Acid detergent ber (ADF), and neutral detergent ber (NDF) were analyzed only in
seeds by methods 942.05, 973.18 (AOAC, 2007), respectively.
The amino acid (AA) content was determined in raw and extruded seeds, diets
and ileal digesta with the assistance of an AAA-400 Automatic Amino Acid Analyzer
(INGOS s.r.o., Praha, Czech Republic) using ninhydrin for post-column derivatiza-
tion. Before analyses, the samples were hydrolyzed with 6NHCl for 24 h at 110°C
(procedure 994.12; AOAC, 2007). Gross energy (GE) was determined in raw and
extruded seeds, diets and excreta using an adiabatic bomb calorimeter (KL 12Mn,
Precyzja-Bit PPHU, Poland) standardized with benzoic acid. Titanium dioxide was
estimated according to Short et al. (1996) and samples were prepared in accordance
with the procedure proposed by Myers et al. (2004).
In the yellow lupine samples, the rafnose family oligosaccharides (RFO) were
extracted and analyzed using high-resolution gas chromatography as described pre-
viously by Zalewski et al. (2001). Phytic phosphorus was determined by extracting
the sample in hydrochloric acid (0.2 M). Next, iron-ammonium sulfate was added
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Raw and extruded lupine seeds in broiler diets 1065
to the centrifuged extract which was heated and then centrifuged. Bipyridine solu-
tion was added to the supernatant and absorbance was determined using spectro-
photometer Media (Marcel Lamidey S.A., Châtillon, France) at 519 nm wavelength
(Haug and Lantzsch, 1983). The water extract viscosity (WEV) of lupin seeds was
measured in vitro. Prior to the determination of WEV, lupin samples were ground in
a mill using a sieve with 0.5 mm mesh and then 1 g of each of the examined cultivars
was mixed with 5 ml distilled water for 1 h at 40°C. The samples were centrifuged at
10 000 g for 10 min at 4°C, the supernatant was withdrawn and viscosity was deter-
mined in a Brookeld Digital DV-II+ cone/plate viscometer (Brookeld Engineer-
ing Laboratories Inc., Stoughton, MA, USA) maintained at 40°C at a shear rate of
12 · s–1. WEV units are mPas·s = cP = 1 × 100 dyne s cm−2
The metabolizable energy of diets used in growth experiment was calculated on
the basis of the European Tables of Energy Values of Feeds for Poultry (1989) and
Smulikowska and Rutkowski (2005).
Calculations of results and statistical analysis
Experiment 1
Using crude fat (CF) calculation as an example, the following equation was used
to calculate the digestibility (AID – apparent ileal digestibility and ATTD – apparent
total tract digestibility) of various dietary components of the basal and experimental
diets:
DC (%) = {1 −[(TiO2% diet / TiO2% digesta/excreta) × (CF%digesta/excreta / CF% diet)]}
The following equation was used to calculate the digestibility coefcients of vari-
ous dietary components and the AMEN level of lupine seeds:
DCCF = (DCCFdiet × CCFdiet–DCCFbasal × CCF basal × 0.20) / (CCFdiet–CCFbasal × 0.20)
where:
DCCF = digestibility coefcient of CF,
DCdiet = digestibility coefcient CF in diet,
Cdiet= concentration of CF in diet,
DCbasal = digestibility coefcient CF in basal diet,
Cbasal = concentration of CF in diet.
AMEN of all basal and experimental diets was calculated using the above equa-
tions and was corrected to zero nitrogen balance using 34.4 kJ/g N retained (Hill and
Anderson, 1958).
All data had been previously explored to discard any possible outliers. Analyses
were performed using the appropriate procedures of SAS Software (2009) (distribu-
tion analyses; outliers were dened as observations whose distance to the location
estimate exceeded 3 times the standard deviation). The obtained results were sub-
jected to the T test, at the level of P<0.05.
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A. Rutkowski et al.
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Experiment 2
The apparent digestibility of nutrients and apparent metabolizable energy value
of diets were determined by the classical method. AMEN was corrected to zero nitro-
gen balance using 34.4 kJ/g N retained (Hill and Anderson, 1958)
Statistical calculations were conducted with the assistance of the SAS® v.9.1
package (2009). Differences between treatments and experimental factors were de-
termined by employing the two-way linear model of ANOVA:
Yijk= αj + βk + (αβ)jk + eijk
where:
Yijk is the value of the analyzed trait,
αj is the constant effect of the ith lupine seed,
βk is the constant effect of the extrusion,
(αβ)jk is the interaction between α and β,
eijk is the effect of experimental error.
Means from experiments were compared using the Duncan’s test and the signi-
cance of differences was assumed to be at the level of P ≤0.05.
Results
Experiment 1
The chemical composition of raw or extruded seeds of yellow lupine (Mister cv.)
was shown in Table 1. Raw seeds have 39% protein and 0.027 g/kg of alkaloids. Ex-
trusion slightly changed the content of basic nutrients, amino acids and anti-nutritive
substances (not conrmed statistically) and reduced the content of phytic phosphorus
in seeds from 0.7 to 0.59 % (Table 1). It should be stressed that measurements were
performed in two replications per sample and were not elaborated statistically.
Table 4. Coefcients of apparent total tract digestibility (ATTD) of dry matter, crude fat, nitrogen
retention and, AMEN value and AMEN/GE of raw and extruded yellow lupine seeds (Experiment 1)
Item Lupine seeds SEM P-value
raw extruded
Dry matter ATTD 0.632 0.649 0.011 0.071
Crude fat ATTD 0.767 B 0.910 A 0.013 <0.001
Nitrogen retention ATTD 0.481 b 0.565 a 0.036 0.021
AMEN (MJ/kg) 9.53 B 10.64 A 0.14 <0.001
AMEN/GE (%) 53 B 57 A 0.080 <0.001
a, b, A, B – within main effects means in columns with different superscripts differ signicantly at:
a, b – P<0.05, A, B – P<0.01.
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Raw and extruded lupine seeds in broiler diets 1067
Extrusion improved total tract and ileal fat digestibility (P<0.001) and the appar-
ent metabolizable energy value of lupine seeds (Table 4). Also, apparent nitrogen
retention in birds fed diets with extruded lupine was higher (P<0.05), compared to
diets containing raw seeds. Birds fed yellow lupine seeds in extruded form had better
ileal digestibility of crude protein and all analyzed AAs, except Lys, Thr and Val and
also crude fat (Table 5).
Table 5. Coefcients of apparent ileal digestibility of crude protein, crude fat and amino acids from
yellow lupine seeds (Experiment 1)
Coefcients Lupine seeds SEM P-value
raw extruded
Crude protein 0.701 B 0.819 A 0.027 0.004
Crude fat 0.712 B 0.843 A 0.027 0.001
Asp 0.838 0.812 0.016 0.087
Thr 0.719 0.758 0.028 0.283
Ser 0.722 b 0.804 a 0.026 0.015
Glu 0.703 B 0.892 A 0.024 <0.001
Pro 0.761 0.789 0.024 0.333
Gly 0.876 A 0.778 B 0.017 0.001
Ala 0.758 B 0.813 A 0.024 0.047
Val 0.802 0.794 0.022 0.704
Iso 0.782 b 0.832 a 0.022 0.047
Leu 0.603 A 0.796 B 0.026 <0.001
Tyr 0.622 a 0.798 b 0.023 <0.001
Phe 0.858 a 0.821 b 0.016 0.045
His 0.822 A 0.602 B 0.020 <0.001
Lys 0.661 0.649 0.014 0.988
a, b, A, B – within main effects means in columns with different superscripts differ signicantly at: a, b –
P<0.05, A, B – P<0.01.
Experiment 2
Extrusion of yellow lupine seeds showed a positive effect on apparent nitrogen
retention. Moreover, there was clear negative effect of dietary level of yellow lupine
seeds on value of this parameter (Table 6). Applying 20, 25 and 30% of seeds in di-
ets decreased the apparent nitrogen retention (P<0.01) (Table 6). The highest crude
fat digestibility was noted for treatments fed diet with 10% of yellow lupine seeds
inclusion (P<0.05). In general, the extrusion signicantly (P<0.01) improved the
total fat digestibility. The AMEN value of the diets was signicantly affected by lu-
pine level. A high ratio of raw lupine (20–30%) clearly decreased the AMEN value
of the diets used, the extrusion of lupine seed improved AMEN by 0.28 MJ/kg
(P≤0.0001).
The health status of chickens was very good, culls and mortality were very low
(2%). Signicant (P<0.01) differences in performance indices among treatments and
also for experimental factors were noted (Table 7). The highest level of yellow lupine
affected negatively (P<0.01) the BWG. Inclusion of 30% of raw or extruded lupine
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A. Rutkowski et al.
1068
caused a depression in feed consumption and also, in growth which was especially
pronounced in chickens fed diets with raw lupine seeds. The feed conversion ratio
in chickens was relatively similar (1.67–1.70 kg/kg) at all dietary lupine levels. An
exception was noted for chickens fed diets with 25% of raw seeds (1.76–1.86) and
when 30% of extruded lupine was incorporated into the diet (2.01 kg/kg BWG).
Baro-thermal processing of lupine seed signicantly decreased the feed intake and
in consequence – also inuenced the FCR for day 15–35 and 0–35 periods. Interac-
tions between lupine levels and extrusion effects were signicant for feed intake and
FCR.
Table 6. Coefcients of total tract apparent nitrogen retention, fat digestibility and apparent metaboliz-
able energy value of diets (MJ/kg) (Experiment 2)
Treatment
Yellow
lupine
%
Extrusion Nitrogen
retention
Crude fat
digestibility AMEN
130 - 0.565 0.893 D 13.64 B
225 - 0.586 0.887 D 13.62 B
3 20 - 0.608 0.896 D 13.76 B
4 10 - 0.643 0.914 BC 14.47 A
5,10 0 - 0.634 0.920 AB 14.45 A
6 30 + 0.630 0.933 A 14.44 A
7 25 + 0.652 0.916 ABC 14.45 A
8 20 + 0.634 0.921 AB 14.19 A
9 10 + 0.662 0.929 AB 14.44 A
SEM 0.004 0.001 0.13
P-value <0.001 <0.001 <0.001
Lupine level (L) 30 0.598 C 0.911 B 14.04 B
25 0.619 BC 0.902 C 14.04 B
20 0.621 BC 0.908 BC 13.98 B
10 0.652 A 0.920 A 14.46 A
0 0.634 AB 0.914 AB 14.45 A
Extrusion effect (E) - 0.612 B 0.903 B 13.98 B
+ 0.645 A 0.923 A 14.26 A
P – level for
L <0.001 <0.001 <0.001
E <0.001 <0.001 <0.001
Interaction L × E 0.147 0.029 <0.001
a, b, A, B – within main effects means in columns with different superscripts differ signicantly at: a, b –
P<0.05, A, B – P<0.01.
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Raw and extruded lupine seeds in broiler diets 1069
Table 7. Performance of broiler chickens (Experiment 2)
Treatments Content
of yellow lupine Extrusion BWG (g) FI (g) FCR
0–14d 15–35 0–35 0–14d 15–35 0–35 0–14d 15–35 0–35
130 - 185 F 1272 1459 381 2061 C 2442 C 2.07 A 1.62 D 1.67 D
225 - 291 E 1511 1802 465 2879 A 3344 A 1.61 B 1.90 B 1.86 B
3 20 - 336 D 1650 1985 478 2864 A 3342 A 1.43 CD 1.74 BC 1.68 CD
4 10 - 348 CD 1626 1973 470 2880 A 3350 A 1.36 DE 1.77 BC 1.70 CD
5,10 0 - 347 CD 1626 1973 480 2857 A 3336 A 1.39 CDE 1.76 BC 1.69 CD
6 30 + 281 E 1313 1596 435 2767 B 3202 B 1.54 BC 2.10 A 2.01 A
7 25 + 371 BC 1531 1902 507 2836 AB 3342 A 1.37 DE 1.85 B 1.76 BC
8 20 + 389 AB 1617 2005 509 2830 AB 3339 A 1.31 DE 1.75 BC 1.67 CD
9 10 + 410 A 1613 2022 520 2884 A 3405 A 1.27 E 1.79 BC 1.68 CD
SEM 6.85 16.01 21.29 5.76 27.52 30.78 0.02 0.02 0.01
P-value <0.001 <0.001 <0.001 <0.001 <0.001 <0.001 <0.001 <0.001 <0.001
Lupine level
30 233 D 1292 C 1527 C 408 B 2414 B 2822 B 1.80 A 1.87 A 1.85 A
25 331 C 1521 B 1852 B 486 A 2858 A 3343 A 1.49 B 1.88 A 1.81 A
20 362 AB 1633 A 1995 A 493 A 2847 A 3340 A 1.36 C 1.74 B 1.67 B
10 379 A 1619 A 1998 A 495 A 2883 A 3377 A 1.32 C 1.78 B 1.69 B
0 349 BC 1604 A 1952 A 480 A 2857 A 3337 A 1.39 C 1.78 B 1.71 B
Extrusion effect - 301 B 1537 1838 455 B 2708 B 3162 B 1.57 A 1.76 B 1.72 B
+ 363 A 1518 1881 493 A 2829 A 3246 A 1.37 B 1.87 A 1.78 A
P-values
Lupine level <0.001 <0.001 <0.001 <0.001 <0.001 <0.001 <0.001 0.039 <0.001
Extrusion effect <0.001 0.879 0.057 <0.001 <0.001 <0.001 <0.001 <0.001 <0.001
Interaction 0.130 0.689 0.439 0.842 <0.001 <0.001 <0.001 <0.001 <0.001
a, b, A, B – within main effects means in columns with different superscripts differ signicantly at: a, b – P<0.05, A, B – P<0.01.
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A. Rutkowski et al.
1070
Discussion
Some harmful factors of legume seeds such as the proteases – trypsin and chymo-
trypsin inhibitors, lectins, oligosaccharides – are sensitive to thermal or baro-thermal
processing such as toasting, granulation, extrusion, boiling and also micronization.
Through such preparation a decrease in their activity or anti-nutritive properties may
be obtained (Leontowicz et al., 2001; Kiczorowska and Lipiec, 2002 a; Kiczorowska
et al., 2002 b). In the present investigations, extrusion of yellow lupine seeds to
a limited degree changed the chemical composition of seeds. Very small differences
in amino acid content between extruded and raw lupine seed were found. A decrease
in phytic phosphorus levels was obtained. It conrms the nding by Alonso et al.
(2000), who explained that during extrusion some molecules of inositol hexaphos-
phate were hydrolyzed to penta-, tetra- and triphosphates; therefore, the level of
phytic P in seeds after extrusion was lower. A similar result was conrmed in our
earlier study for faba bean and pea (Hejdysz et al., 2016 a, b).
Nevertheless, the apparent nitrogen retention, total digestibility of fat and the
AMEN value of lupine seeds determined in young chicken show a signicant bene-
cial effect of extrusion of yellow lupine. Also signicantly better ileal digestibility of
protein and some amino acids from extruded seeds was found in comparison to raw
lupine seeds. A similar benecial effect of extrusion in terms of protein digestibility
in rats was reported by Leontowicz et al. (2001).
Increased levels of lupine seeds in broiler diets, both raw or extruded seeds, led
to numerous signicant differences among treatments. Using 10 to 30% of lupine,
the apparent nitrogen retention, fat digestibility and AMEN value of diets was sig-
nicantly better in birds fed diets with extruded lupine, while apparent nitrogen
retention (63–66% of N intake), fat digestibility (about 92%) and AMEN value
(14.19–14.44 MJ/kg) were relatively similar in these treatments.
By the addition of 10 and 20% of lupine in diets, positive effects of extrusion
were noted for BWG, FI and feed conversion ratio. Use of 25%, especially 30% of
lupine, decreased the performance indices. Extrusion to a limited degree decreased
the negative action of large amounts of lupine seeds on broiler growth.
The obtained performance results are in agreement with the experiments of other
authors (Leontowicz et al., 2001; Orda et al., 2006). The use of raw as well as ex-
truded yellow lupine seeds at 10 to 20% of the diet, can be expected to lead to
a benecial effect. However, with higher levels (25%) of these seeds in broiler diets
the baro-thermal processing of lupine may be benecial.
The poorer performance results registered for feeding 30% and also 25% of raw
as well as extruded yellow lupine seeds indicate that such levels of these legume
seeds are too high for young broiler chickens. However, the introduction of 25% of
extruded yellow lupine made it possible to obtain better results than with raw seeds.
The use of 10 or 20% of raw or extruded lupine in the broiler diets resulted in similar
effects in terms of growth and feed conversion, compared to the diet that included
soybean meal as a main protein source.
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Raw and extruded lupine seeds in broiler diets 1071
Conclusions
The extrusion process increased the price of lupine seeds, as a result of the en-
ergy costs involved in baro-thermal processing, but it could be benecial when high
yellow lupine meal inclusion is applied. The diet containing 10 or 20% of raw and
extruded yellow lupine allows obtaining similar results as in control.
References
A l o n s o R., A g u i r r e A., M a r z o F. (2000). Effects of extrusion and traditional processing methods
on antinutrients and in vitro digestibility of protein and starch in faba and kidney beans. Food Chem.,
68: 159–165.
AOAC (2007). Ofcial Methods of Analysis of the Association Ofcial Analytical Chemists. Arlington,
USA.
E r b a ş M., C e r t e l M., U s l u M.K. (2005). Some chemical properties of white lupin seeds (Lupinus
albus L.). Food Chem., 89: 341–345.
European Table of Energy Values for Poultry Feedstuffs (1989). WPSA, Beekbergen, The Netherlands.
G d a l a J. (1998). Composition, properties and nutritive value of dietary bre of legume seeds. A re-
view. J. Anim. Feed Sci., 7: 131–149.
Guillamon E., Pedrosa M.M., Burbano C., Cuadrado C., De Cortes Sanchez M.,
M u z q u i z M. (2008). The trypsin inhibitors present in seed of different grain legume species and
cultivar. Food Chem., 107: 68–74.
H a u g W., L a n t z s c h H.J. (1983). Sensitive method for the rapid determination of phytate in cereals
and cereal products. J. Sci. Food Agric., 34: 1423–1426.
H e j d y s z M., K a c z m a r e k S.A., R u t k o w s k i A. (2015). Factors affecting the nutritional value
of pea (Pisum sativum) for broilers. J. Anim. Feed Sci., 24: 252–259.
H e j d y s z M., K a c z m a r e k S.A., R u t k o w s k i A. (2016 a). Extrusion cooking improves Faba
been seed metabolic energy and amino acid digestibility for broiler chickens. Anim. Feed Sci. Tech.,
212: 100–111.
H e j d y s z M., K a c z m a r e k S.A., R u t k o w s k i A. (2016 b). Effect of extrusion on the nutritional
value of peas for broiler chickens. Arch. Anim. Nutr., 70: 364–377.
H i l l F., A n d e r s o n D.L. (1958). Comparison of metabolizable energy and productive energy deter-
mination with growing chicks. J. Nutr., 64: 587–593.
J u l L.B., F l e u g m a r k P., G u ll i ng M., I t e n o w K. (2003). Lupin seed (Lupinus albus and Lupi-
nus luteus) as protein source for fermentation use. Indust. Crop. Prod., 18: 199–211.
Kaczmarek S.A., Kasprowicz-Potocka M., Hejdysz M., Mikula R., Rutkowski A.
(2014). The nutritional value of narrow-leafed lupin (Lupinus angustifolius) for broilers. J. Anim.
Feed Sci., 23: 160–166.
K a c z m a r e k S.A., H e j d y s z M., K u b i s M., R u t k o w s k i A. (2016). Inuence of graded in-
clusion of white lupin (Lupinus albus) meal on performance, nutrient digestibility and intestinal
morphology of broiler chickens. Brit. Poultry Sci., 57: 364–374.
K i c z o r o w s k a B., L i p i e c A. (2002 a). The quality of protein in lupine extrudates as affected by
extrusion-cooking parameters (in Polish). Acta Scient. Polon. Techn. Aliment., 1: 83–91.
K i c z o r o w s k a B., L i p i e c A. (2002 b). The quality of fat in lupine extrudates according to the
parameters of extrusion – cooking (in Polish). Acta Scient. Polon. Techn. Aliment., 1: 93–99.
K l u g e H., H i r ch e F., E d e r K. (2002). Concentrations of NSP and oligosaccharides in lupine spe-
cies L. angustifolius, L. luteus and L. albus. Tag. Schweine Geugelernaehrung, Lutherstadt Wit-
tenberg, pp. 145–147.
Krawczyk M., Mikulski D., Przywitowski M., Jankowski J. (2015). The effect of di-
etary yellow lupine (L. luteus cv. Baryt) on growth performance, carcass characteristics, meat qual-
ity and selected serum parameters of turkeys. J. Anim. Feed Sci., 24: 61–70.
Leontowicz H., Leontowicz M., Kostyra H., Kulasek G., Gralak M.A., Krze-
m i ń s k i R., P o d g u r n i a k M. (2001). Effects of raw or extruded legume seeds on some func-
tional and morphological gut parameters in rats. J. Anim. Feed Sci., 10: 169–183.
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A. Rutkowski et al.
1072
M a ń c z a k T., D o l a t a A., Wi a t r K. (2007). The contents of quinozidine alkaloids and gramine
in seeds of lupin cultivars registered in Poland (in Polish). Zesz. Probl. Pods. Nauk Rol., 522:
463–471.
M y e r s W.D., L u d d e n P.A., N a yg i hu g u V. B., H e s s W. (2004). Technical note: A procedure
for the preparation and quantitative analysis of samples for titanium dioxide. J. Anim. Sci., 82:
179–183.
N a l l e C.L. (2009). Nutritional Evaluation of Grain Legumes for Poultry. Ph.D. Thesis. Massey Uni-
versity, Palmerston North, New Zealand.
O l k o w s k i B.I., C l a s s e n H.L., W o j n a r o w i c z C., O l k o w s k i A.A. (2005). Feeding high
levels of lupine seeds to broiler chickens: plasma micro nutrient status in the context of digesta
viscosity and morphometric and ultra structural changes in the gastrointestinal tract. Poultry Sci.,
84: 1707–1715.
Orda J., Jamroz D., Wiliczkiewicz A., Wertelecki T., Skorupińska J., Broz J.
(2006). Effects of increased dietary inclusion of yellow lupins and enzyme supplementation on per-
formance, ileal digestibility of nutrients and microbial status of large intestine in broiler chickens.
Arch. Geugelkd., 70: 14–21.
Rutkowski A., Kaczmarek S., Hejdysz M., Nowaczewski S., Jamroz D. (2015).
Concentrates made from legume seeds (Lupinus angustifolius, Lupinus luteus and Pisum sativum)
and rapeseed meal as protein source in laying hen diets. Ann. Anim. Sci., 15: 129–142.
S h o r t F.J., G o r t o n P., W i s e m a n J., B o o r m a n K.N. (1996). Determination of titanium dioxide
added as an inert marker in chicken digestibility studies. J. Anim. Feed Sci. Tech., 59: 215–221.
Smulikowska S., Pastuszewska B., Mieczkowska A., Lechowski R. (1999). Effect
of gramine – indole alkaloid of yellow lupin on performance and some physiological parameters in
broiler chicken. Proc. 12th Europ. Symp. on Poultry Nutrit., Veldhoven, pp. 313–315.
S m u l i k o w s k a S., R u t k o w s k i A. (2005). Editors. Recommended allowances and nutritive value
of feedstuffs – poultry feeding standards, 4th edition (in Polish). The Kielanowski Institute of Ani-
mal Physiology and Nutrition, PAS, Jablonna, Poland.
S u j a k A., K o t l a r z A., S t r ob e l W. (2006). Compositional and nutritional evaluation of several
lupin seeds. Food chem., 98: 711–719.
W a s i l e w k o O.J., B u r a c z e w s k a L. (1999). Chemical composition including content of amino
acids, minerals and alkaloids in seeds of three lupin species cultivated in Poland. J. Anim. Feed Sci.,
8: 1–12.
Z a l e w s k i K., L a h u t a L.B., H o r bo w ic z M. (2001). The effect of soil drought on the composi-
tion of carbohydrates in yellow lupine seeds and triticale kernels. Act. Physiol. Plant., 23: 73–78.
Zduńczyk Z., Jankowski J., Mikulski D., Mikulska M., Lamparski G., Slomins-
k i B.A., J u ś k i e w i c z J. (2014). Growth performance, gastrointestinal function and meat quality
in growing nishing turkeys fed diets with different levels of yellow lupine (L. luteus) seeds. Arch.
Anim. Nutr., 68: 211–226.
Received: 17 II 2015
Accepted: 4 IV 2016
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