The Effect of Lupine Seed in Broiler Diet on Animal [601665]

The Effect of Lupine Seed in Broiler Diet on Animal
Performance and Fatty Acids Profile of their Meat
Daniel MIERLITA
Department of Animal Science, University of Oradea, 26Magheru street, Romania;
*Corresponding author, e-mail: [anonimizat]
Bulletin UASVM Animal Science and Biotechnologies 72(2) / 2015
Print ISSN 1843-5262; Electronic ISSN 1843-536X
DOI:10.15835/buasvmcn-asb:11375
Abstract
Alkaloid-free lupine seed may represent an alter native for ensuring the vegetable protein in poultry feed.
Given the weight but especially the place and role of fats in maintaining consumer health, in our research we
watched the effect of alkaloid free lupine seed in the diet of broiler (chicken and turkey), the animal performance
and especially the effect on the fatty acids composition in the chest intramuscular fat. The experiment was
conducted as a completely randomized experimental design consisting of two treatments involving a control diet
consisting of corn – soybean grits (LC) and an experimental diet (LE) in which the proteins from soy grits were
replaced by alkaloid free lupine flour. Two groups were organized: LC (control) – in whose diet soybean meal was
used and LE protein (experimental) – in whose diet alkaloid free lupine seeds were used. In the chicken broiler diet,
the protein derived from soybean meal was replaced with alkaloid free lupine flour in proportion of 30% during
days 1-21 and 60% during days 22-42 (14.0-25.7% lupine ratio in the combined fodder structure). As for the
turkey broiler, lupine seed accounted for 30% of combined fodder structure, substituting between 40% (breeding
stage 0-3 weeks) and 75% (breeding stage 15-18 weeks) of protein provided by soybean meal. Substitution of
soybean meal protein in the broiler diet by introducing alkaloid free white lupine beans in broiled food has no
negative effect on their bio-performance. Supplementing the broilers diet with lupine flour resulted in a decreased
share of saturated fatty acids with high atherogenic potential (C14:0, C16:0, C18:0) in breast intramuscular fat
and increased the share of n-3 series polyunsaturated fatty acids (α-linolenic acid especially), which at their turn
decreased the n-3/n-6 series fatty acids in breast, thus increasing the nutritional quality of poultry meat, analysed
in terms of impact on human consumer health.
Keywords : alkaloid-free lupine, fatty acids composition, meat, broiler.
INTRODUCTION
Using white lupine seed free of alkaloids
in broiler food is justified by the high content of
protein (40.08% CP), but also fat (9.70% crude
fat) rich in polyunsaturated fatty acids (PUFA)
consisting mainly of linoleic acid (C18 : 2n-6) and
linolenic acid (C18: 3n-3) (Mierliță, 2012; 2013;
2014).
The use of white lupine seeds in proportions
exceeding 25% (% of fodder structure) does not
affect the production performance of broiler (Nalle
et al ., 2010; Suchy et al ., 2010). The use of lupine
seeds in bird feeders, as the sole source of protein,
is limited on the one hand by the biological value of the feed in terms of less protein content (low in
amino acids limits) and on the other hand by the
high content in NSP (non-starch polysaccharides)
(Strakova et al ., 2006; Mieczkowski et al ., 2004).
White lupine seeds can substitute 50% of the
protein provided by soybean meal in broiler
chickens if seeds are shelled or if the diet is
supplemented with synthetic amino acids and
enzymes (Suchy et al ., 2006).
Research has primarily aimed at determining
the effect of the introduction of white lupine seeds
free of alkaloids in broiler chickens (chickens and
turkeys) on productive parameters but mainly on

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fatty acid composition of the intramuscular fat in
the chest.
MATERIALS AND METHODS
Two diets were tested: LC (control) – standard
based diet soybean meals and LE (experimental) –
diet based on white lupine seeds free of alkaloids.
In the chicken broiler diet, the protein derived
from soy grits was replaced by alkaloid free lupine
flour in proportion of 30% during days 1-21 and
60% during days 22-42 (14.0-25.7% lupine ratio
in the combined fodder structure). As for the
turkey broiler, lupine seed accounted for 30% of
combined fodder structure, substituting between
40% (breeding stage 0-3 weeks) and 75%
(breeding stage 15-18 weeks) of protein provided
by soybean meal.
White lupine seeds (variety Energy) came
from own culture made by SCDA Oradea (Mierlita,
2012). Gross chemical composition and energy
value analyses were performed in our laboratory,
using specific established methods (AOAC, 1996;
Sibbald 1980). Lupine seeds were analysed for
the content of total alkaloids as described by Ruiz
(1977).
Diets were isoenergetic and isonitrogenous,
including limiting amino acids (lysine and
methionine). Mixed fodder was given in plenty, the
amount of feed consumed being determined for
each boilers lot and for each period of breeding,
respectively, by means of weighing.
In order to assess the quantitative and
qualitative indicators of meat production, eight
broilers per each lot were randomly selected for
slaughter purposes. The raw chemical composition
of breast meat was determined using the Weende
method.At slaughter, samples were collected from
every 150-200 g muscle tissue of the breast
(skinless) and stored at -200 C until laboratory
analysis on the fatty acid composition of the
intramuscular fat. Lipid extraction was done in a
mixture of chloroform / methanol (2:1, vol/vol).
Fatty acid methyl esters (FAME) were obtained
from intramuscular lipids using the method
described by Watkins et al . (1997), using boron
trifluoride (BF3) and methanol (14% w/w –
USA). Methylation was carried out with sodium
methoxide. Dissolution of lipids extracted in 1
ml of toluene was done in test tubes with teflon
lid. In order to obtain FAME 0.5 ml of sodium
methoxide and 2 ml anhydrous methanol were
added, the mixture was maintained for 10 min
at 50°C, adding 0.1 ml of glacial acetic acid and
5 mL of deionized water. To identify FAME, a gas
chromatograph (GC HP 5890 Series II) with a
capillary column: 30m x 0.53 mm i.d was used.
The gas chromatographic operating parameters
were: 1400 C for 2 min; 1.50 C/min up to 2000 C
and held for 6 min. Injector and detector flame
ionization indicated temperatures 2250 C and 2600
C, respectively. FAME identification was made by
their retention times with standards (not-Check-
Prep., USA). Expression of fatty acids was made in
percentage of the total FAME identified.
The obtained data were subjected to
variance analysis by using ANOVA (Pallant,
2007). Comparisons among means were done
with the Tukey test. The level of significance was
established at P ≤0.05.
RESUL TS AND DISCUSSION
Lupine beans have a higher crude protein
content (44.00% in DM), crude fat (10.69% in DM)

Tab. 1. Gross chemical composition and energy value lupine beans (% of DM –dry matter)

Mean Max. Min.
Crude protein 44.00 44.90 41.93
Crude fat 10.69 13.27 9.59
Crude fiber 16.10 17.46 14.31
SEN (non-nitrous
extractables)24.92 26.51 22.38
Crude ash 4.00 4.94 3.66
Mean (kcal/kg) * 3326.9 3482.3 3158.6
Alkaloids (mg / kg) 110.6 174.4 96.7
* calculated according to Sibbald, 1980.The effect of lupine seed in broiler diet on animal performance and fatty acids profile of their meat

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Bulletin UASVM Animal Science and Biotechnologies 72(2) / 2015 MIERLITA et al
and crude fiber (16.10% in DM). Due to their high
fat content, lupine seeds have a high energetic
value (3326.9 MEAn/kg DM) (Table 1). High crude
fiber content of lupine seeds was confirmed by
Brenes et al . (1993). Husking white lupine seeds
may increase the bioavailability of nutrients so
that the nutritional value of lupine seeds could be
comparable to soybean meal (Suchý et al ., 2010).
Fats of lupine beans are high in long chain
fatty acids, better represented by oleic acid (C18:
1; 40.05% of the total identified fatty acids),
linoleic acid (C18: 2 n-6; 24, 81% of the total FA
identified) and α-linolenic acid (C18:3 n-3; 8.88%
of the FA identified) (Table 2). Green and Oram
(1983) set the content of lupine seeds at approx.
50% oleic acid, 22% linoleic acid and approx. 10%
α-linolenic acid; our results confirm these values.
The results obtained in these studies are
consistent with those obtained in other studies
(Rubio et al ., 2003; Erbas et al ., 2005; Sujak et al .,
2006; Uzun et al ., 2007; Vecerek et al ., 2008) and
confirm that the lupine seeds are an important
source of protein for animals, and of unsaturated
fatty acids, which can help improve the fatty acid
composition of the meat.The results show that when mixed fodder is
properly balanced in terms of energy, protein and
limiting amino acids, substituting soybean meals
protein in broilers diet with alkaloid free white
lupine flour has no negative effect on weight gain,
the level of food use, carcass quality and meat (see
Table 3). Compared with the control group, the
final weight of the chickens in the experimental
group was lower by approx. 4%, feed intake was
lower by approx. 2.7%, feed consumption for kg
gain in weight was higher by approx. 1.2% and the
chest carcass was lower by approx. 0.3 percentage
points. However the differences were not
statistically assured. The introduction of lupine
in broiler chickens had lowered the price of a kg
fodder up to 18%, while feed costs per 1 kg gain
that weight dropped to 17%. If the diet is balanced
in amino acids (lysine, methionine, tryptophan),
introduction of white lupine seeds free of alkaloids
in broiler chickens (chickens and turkeys) in the
amount of max. 250 g/kg fodder does not affect
their productive performance (Roth-Maier and
Paulicks, 2003). Special attention should be paid
to sulfur amino acids that are deficient in white
Tab. 2 . Fatty acid composition of lupine seed (% of FAME)
Fatty acid X mean X min. X max.
C12:0 0.78 0.63 1.07
C14:0 0.15 0.08 0.23
C16:0 6.79 4.30 11.64
C16:1 1.03 0.34 1.73
C18:0 4.60 2.58 6.93
C18:1n-9 40.05 36.20 49.61
C18:2n-6 24.81 22.19 30.98
C18:3n-3 8.88 6.36 12.14
C20:0 1.25 0.99 1.69
C20:1n-9 6.57 6.20 8.07
C20:2n-6 1.15 0.81 1.72
C20:4 3.94 2.83 4.88
Total SFA 17.51 8.58 21.56
Total MUFA 47.65 42.74 59.41
Total PUFA 34.84 32.19 49.72
PUFA n-6 25.96 23.00 32.70
PUFA n-3 8.88 6.36 12.14
n-6/n-3 FA 2.92/1 3.61/1 2.69/1
FAME = fatty acid methyl esters; FA = fatty acid; SFA = saturated fatty acid; MUFA = monounsaturated fatty acid; PUFA =
polyunsaturated fatty acid.

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lupine seeds, representing only 1/3 of that found
in soybean meal (Nalle et al ., 2010).
The negative influence of lupine is attributed
to the large amounts of NSP (non-starch
polysaccharides). They have a negative effect on
both feed intake and use (Roth-Maier and Paulics,
2003; Steenfeldt et al ., 2003).
Partial replacement of soybean with lupine
flour in broilers (chicken and poultry) resulted in
no significant differences (p >0.05) in the chemical
composition of breast muscles. The only exception
is the crude ash content which has significantly
increased (p<0.01) in chickens compared to the
control lot (Lc). Our results are in agreement
with those obtained by Suchy et al . (2010), who
reported that replacing soy nitrogen at a rate of
up to two-thirds does not affect the protein and
fat content in breast and thigh muscles, but it
only increases the crude ash content in the breast.
Contrary to our results, Suchy et al . (2010) found a
drop in crude ash content of thigh muscle.
Intramuscular fat fatty acid profile of breast
has been significantly improved by the introduction
of lupine seeds in broiler diet. Thus, compared
with the control group there was an increase in
the share of monounsaturated fatty acids (MUFA)
and especially PUFA and decrease of saturated
fatty acid (SFA). Stearic acid (C18: 0), which has
negative effects on the cardiovascular consumers,
recorded the largest drop in the structure of chest
fat in chickens which were introduced white lupine
seeds (Table 4).
The largest share of PUFA is represented
by the linoleic acid (LA, C18:2n-6), but the most important one, analysed in terms of impact on
the consumer’s health, is the share of α-linolenic
acid (ALA, C18:3n-3). Placing lupine flour in the
broilers’ diet caused an increase of the share of
fatty acids of both the n-6 series and the n-3 series,
in the breast intramuscular fat structure, which is
equivalent to an improvement of their nutritional
features. Thus, compared to the control group (LC),
group LE, weighted LA structure of intramuscular
fat increased from 54.4%, while ALA share rose
from 91.04%. The highest increases were recorded
in turkey (Mierlita, 2013). It is worth noting that
the highest content of C18: 2 n-6 (omega-6) was
recorded in broilers, while broiler turkeys showed
the highest concentration of C18: 3 n-3 (omega 3).
These results demonstrate the superior quality
of the turkey meat, from a nutritional standpoint,
determined on the one hand by the content of lower
fat breast meat, and on the other hand the higher
content of intramuscular fatty acid Omega-3.
Increasing the share of n-3 series polyunsaturated
fatty acids in meat, as a result of including lupine
flour in broilers’ (chickens and poultry) diet is
particularly beneficial to consumer’s health, as
the nutritional qualities of poultry meat (and
especially of turkey) are improved.
Increasing the share of n-3 FA and narrowing
ratio n-6 / n-3 FA is beneficial for human health.
However, in the case of broiler chickens ratio
n-6 / n-3 was higher than that recommended
by nutritionists for healthy human nutrition 4:1
(Nalle, 2009).
Similar studies have shown that white
lupine seeds placed in diets lead to increased Tab. 3.The effect of lupine seed in broiler diet on growth performance
VariableChicken broiler Turkey broiler
Lc Ec Lc Et
Body weight
(g)Initial 41.6 ± 0.67 41.9 ± 0.49 50.7 ± 0.23 50.4 ± 0.53
Final 2598.7 ± 42.9 2493.8 ± 48.16 19.87 ± 0.28119.07 ± 0.25
Average daily gain (g) 60.85 58.36 157.29 150.95
Feed intake (g/day) 108.63 105.49 419.0 405.9
FCR (kg feed/kg gain) 1.785 1.807 2.664 2.689
Costs feed (Ron/kg gain) 2.77 2.27 4.29 3.83
Slaughter yield
(%)Carcass gril 68.65 ± 4.73 67.07 ± 3.72 72.47 ± 0.57 72.54 ± 0.72
Commercial 75.69 ± 7.31 74.16 ± 6.18 74.71 ± 0.62 74.64 ± 0.64
Quality of the
carcass (%)Breast 35.72 ± 1.71 35.43 ± 0.43 37.08 ± 2.05 36.43 ± 0.77
Hams + Thigh 30.81 ± 1.10 30.72 ± 0.84 17.59 ± 0.67 18.74 ± 0.52
Lc – control group; Ec – experimental chicken group; Et – experimental turkey group; FCR – feed conversion ratio; 1 – kg/turkey broilerThe effect of lupine seed in broiler diet on animal performance and fatty acids profile of their meat

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concentration of omega-3 FA (especially C18: 3n-
3) in poultry fat (Strakova et al ., 2006). Zelenka
et al . (2008) considers that supplementing the
broilers’ diet with fat rich in essential fatty acids
provides better quality meat.
Including white lupine seeds in broiler food
had positive effects on meat products resulting
from the metabolism of PUFA, especially of
linolenic acid: docosapentaenoic acid (DPA, C22:
5n-3) and eicosapentaenoic acid (EPA, C20: 5n-3).
Their share in the structure of intramuscular fat
increased by 35.2-62.6% for chickens and 47.2-
76.6% in turkey (p <0.01) (Mierlita, 2013). Our
results are supported by research conducted by
Nalle (2009), who demonstrated that there is a
positive correlation between the level of C18: 3n-3
in broiler chickens and concentration of EPA and
DPA in intramuscular and intra-abdominal fat.
CONCLUSION
Use of lupine seeds free of antinutritive
substances in broiler diets did not affect the final weight, degree of recovery of the food or the quality
of housing, instead it contributes to reduced costs
for feeding birds. Lupine seeds have improved
the nutritional quality of intramuscular fat in the
chest, resulting in decreased weight of saturated
fatty acids (in particular: C14:0, C16:0, C18:0)
and increasing the share of polyunsaturated fatty
acids n-3 (in particular: C18:3n-3, C20:5n-3 and
C22:5n-3) and n-6 (in particular: C18:2n-6). The
ratio n-6 / n-3 FA narrowed, making the meat
favourable for human nutrition.
Acknowledgments. This work was supported
by a grant of the Romanian National Authority
for Scientific Research, CNDI–UEFISCDI, project
number PN-II-IN-CI-2012-1-0257 and project
number PN-II-IN-CI-2013-1-0080.
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Chicken broiler Turkey broiler
Lc Ec Lc Et
Crude fat (%) 8.61 8.47 2.85 2.66
Lauric, C12:0
Miristic, C14:0
Pentadecanoic, C15:0
Palmitic, C16:0
Stearic, C18:0
Total SFA0. 12
0.47
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