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EFFECT OF ADDING DIFFERENT LEVELS OF ANNATTO SEED POWDER (BIXA
ORELLANA) IN LAYING CHICKEN DIETS ON OXIDATION INDICATORS
Aseel J. Mohammed1,2, Sunbul J. Hamodi1 and Firas M. H. Alkilani3
1Department of Animal Production, Faculty of Agricultural, University of Baghdad, Iraq.
2Department of Planning and Follow up, Ministry of Agriculture, Iraq.
3Department of Agricultural Research, Ministry of Agriculture, Iraq.
(Accepted 18 July 2018)
ABSTRACT : The present experiment was conducted to estimate the effect of adding different levels of Annatto seeds
(Bixa orellana) (BO) to the laying chicken diets as antioxidants on oxidation indicators in the egg yolk. Two hundred
forty, ISA Brown laying hensat the age of 67-week-old(producing brown eggshell) were employed in this experiment.
The hens were weighed individually and the hens with weights above and below the standard weights (± 10)
wereexcludedaccording to the index of producing company (ACE Brown standard index), randomly and homogeneously
distributed into four treatments by three replicates per treatment (20 hens per replicate). The experimental treatments
included: T1 basal diet as control, T2 basal diet +1% of the BO powder, T3basal diet + 2% of the BO seed powder, T4
basal diet + 3% of the BO seed powder. The obtained results of the present study indicated that adding 1, 2, 3% of BO
seed powder to the diet resulted in a significant difference (P <0.05) between treatments T2, T3 and T4 compared to
T1in the oxidation indicators and for all periods of the experiment. The lowest ratio was observed in T4 with a decrease
in the levels of free fatty acids (FFA), peroxidevalue (PV) and the amount of dehydrogenate (TBA). It can be concluded
that adding Annatto seed powder (BO) seed powder at levels of 1, 2, and 3% to laying chicken diets improved oxidation
indicators in the egg yolk.
Key words : Annatto, oxidation indicators, chicken whites, free fatty acids, peroxide value.
INTRODUCTION
Poultry breeding has expanded tremendously
throughout the world over the past two decades as a result
of the trend of breeders to the intensive system, and the
great technological and genetic advances that have been
developed to meet the needs of consumers (Araya et al,
1977). As a result of this expansion in poultry breeding,
the production of eggs has increased significantly and
raised rapidly as a result of the evolution of methods of
education and health care and the continuous improvement
in the quality and composition of feed in addition to the
development of strains known for their ability and rapid
growth (Shawah, 2009). The nutritional value of eggs
has long been recognized and still occupies an important
nutritional status throughout the world. For this reason,
the countries of the world are seeking to raise their egg
production rates to increase the daily consumption of eggs
as well, to ensure a high level of food for their populations.
Eggs contain all the nutrients that body need, including
carbohydrates, fats, vitamins and minerals. The large egg
contains about 70 calories, 60 of which are concentrated
in egg yolk (Zuhairi, 2011).
The most important problem that faced laying hens
is the oxidative damage and the first sign of this problem
is the sudden increase in the death of hens during the
peak production period, which may reach up to 5%, as
well as the decrease in egg production. There are also
many additional factors that contribute to the occurrence
of oxidative damage such as nutrition, environment,
heredity as well as toxic substances and rancidity of
unsaturated fats in poultry feed in the absence of
antioxidants in the diet,which leads to an increase in the
level of free radicals and therefore Increased oxidation is
offset by weakness in the antioxidant defense system
leading to the accumulation of lipid peroxidation products
in the liver cells. Thus, damage to the hepatic cell
membranes of the membranes of the internal structures
of the cell and the continued excessive increase in oxidative
reactions can cause damage to the biological membranes
(Christaki, 2012). Therefore, the feeding of laying
chickens should be supplemented with the addition of
natural substances such asBOseeds to the feed to enhance
their role as antimicrobials and antioxidants. Annatto
seeds ranked second in economic importance worldwide
among all natural colors and were considered a food factor
Biochem. Cell. Arch. Vol. 18, No. 2, pp. 1621-1624, 2018 www.connectjournals.com/bca ISSN 0972-5075
that did not play a role in taste change but only it is role
as a tincture for egg yolk, where the seeds of BO were
used to feed the laying chickens as a source of coloring
for egg yolks. Most breeders prefer to use these seeds
because they are natural pigments that do not increase
the cost of feed processing and it has a role as anti-
microbial and anti-oxidant (Nicolle et al, 2003; Yolmeh
et al, 2014).
MATERIALS AND METHODS
This experiment was conducted at the Poultry
Research Station of the Livestock Research Department
at the Department of Agricultural Research, Ministry of
Agriculture - Abu Ghraib for the period from the 23 of
October 2017 until the 23 of January 2018 (12 weeks).
In this experiment, two hundred forty ISA Brown chicks
at the age of 67-week (producing brown eggshell) were
used. The chickens received full health care and according
to the preventive health program of the chickens during
the growth stage and according to the recommendations
in the guide of laying chickens. The birds were reared in
cages (60 cm × 60 cm)with a closed-system hall 28 m ×
9 m (length × width), each replicate included 4 cages (per
cage 5 hens). The cages were equipped with the automatic
nipples of the cages for drinking water, and each cage
had three nipples installed at the top of the cage in addition,
each cage was equipped with one feeder. The lighting was
used for 16 hours/day and 8 hours of darkness for the
entire duration of the experiment. A special timer watch
was used to control the lighting and the temperature was
controlled during the winter using electric heaters for the
poultry field. As for the ventilation, the field was equipped
with air exhausts in front of the desert cooling windows.
Moreover, the diet was prepared for successive periods
and the duration of each preparation was 3 weeks to
maintain the effectiveness of the Annatto seeds (BO) added
to the feed to follow the instructions of the company
producing it. The diet elements were grinded by an electric
grinder and a quantity of the specific BO seed powder
was taken for each treatment and mixed with both the
protein concentration, di calcium phosphate and limestone
to ensure uniform, homogeneous mixing gradually until
mechanically mixed by a vertical mixer in the feed plant
of the Agricultural Research Department. The laying
chickens were fed with a uniform diet according to the
requirements of the ISA Brown strain according to it is
guide instructions during the ages of 67-70, 71-74 and
75-78 weeks as shown in Table 1. The feed was provided
by hand and at fixed intervals (120 g/day). The experiment
was carried out according to the complete random design
(CRD). Significant differences between the averages were
measured using the Duncan (1955) test.
RESULTS AND DISCUSSION
Table 2 demonstrated that free fatty acids were
reduced by increasing the concentration of the BO seeds
in the diet compared to the control treatment for the periods
after 4, 8 and 12 weeks of the experiment and recorded
the lowest value of free fatty acids in T2, T3 and T4
compared to the T1 that recorded the highest value of
free fatty acids where the relationship was inverse between
the amount of BO and free fatty acids, the higher amount
of BO combined with the lower ratio of free fatty
acids.BO seeds have anti-microbial properties and can
kill many pathogens and bacteria in the body because
they contain powerful antioxidants and are veryeffective
against many foodborne diseases. Oxidative rancidity is
a major cause of the deterioration of food quality, leading
to unpleasant odors.Synthetic antioxidants like hydroxyl
toluene butyl and hydroxyyannizole are often used
extensively for many years to delay lipid oxidation, but
there has been growing concern about the safety of
1622 Aseel J. Mohammed et al
Table 1 : Chemical composition and diet content of the experiment.
Feed content Percentage %
Yellow corn 40.3
Wheat 30
Soybean meal 48% protein 14
Protein concentration15
Dicalcium phosphate 0.9
Limestone 9.5
Salt 0.3
Total 100
Chemical composition2
Kcal/kg 2734
Crude protein % 15.6
Crude fiber % 2.4
Crude fat % 2.55
Lysine % 0.80
Methionine 0.35
Methionine +cysteine % 0.62
Arginine % 0.54
Calcium % 4.14
Phosphorus % 0.53
1Proteins Center: Proteins Center LAYCON-5 SPECIAL W. Each
containing 40% raw protein, 5% raw fat, 2.15% raw fiber, 7.75%
moisture, 4.30% calcium, 2.00% phosphorus, 25.47 ash, 2.00%
phosphorus, 3.75% Methionine, 0.44% tryptophan, 2.00%
threonine, 1.70% valentine, 3.28% methionine + cysteine, 2.20%
sodium, 2158 kcal / kg energy, 3.00% chloride, 2.59% arginine, ,
500 mg vitamin B, 500 mg vitamin B, 600 mg niacin, 5 mg sodium,
15 mg folic acid, 180 mg calcium pantothenate, 6 mg Choline
Chloride, 5.205 mg Choline, 1.6 mg mg, 20 mg IUD, 1 mg biotin,
1 mg iron, 200 mg copper, 1.6 mg manganese, 1.2 mg zinc, 2.8 mg
propyl kalate, 5 mg citric acid, 33.5 mg antioxidant (BHT),
2According to the chemical analysis of the mixture according to
NRC (1994).
Effect of adding different levels of annatto seed powder in laying chicken diets 1623
synthetic antioxidants (Shahidi and Zhong, 2010).
Therefore, in terms of food security consumers prefer the
use of natural antioxidants like Anatto seeds (BO), which
consideredas a natural source and secure for human
consumption when added to the chicken diets because it
contains resinous fat (6.3%), that has a pungent smell
and warm. Seeds were known for their medicinal
properties, such as anti-inflammatory and anti-oxidant
activity (Prabhakara et al, 2015). Laying chicken is known
to be very effective in transporting fat-soluble food to
egg yolk. This fact provides a way to change the nutritional
composition of eggs (Walker et al, 2012) as a means of
human nutrition. BO seeds were also a good source of
vitamin E which has a role in the biological system as an
oxidation inhibitor through breaking the chain of root
reactions and pulling free radicals. Therefore, the seeds
of BO are a good source of vitamin E that concentrated
in the egg yolk and represents an important element as an
antioxidant as the oxidative stress is currently the leading
theory of many chronic diseases (Shokolenko et al, 2014).
The peroxide value was decreased by increasing the
concentration of the BO seeds in the diet for the periods
after 4, 8 and 12 weeks of the experiment (Table 3). The
lowest values of peroxide in treatments that supplemented
withBO seeds (T2, T3 and T4) compared to the T1, which
recorded the highest value, where the relationship was
inverse between the level of the BO seeds and the peroxide
value whereas the higher level of BOseeds in the diet
represents the lower peroxide value. The above results
indicated that the addition ofBO seeds to the diet reduces
the fat deterioration inegg yolk resulting in lower peroxide
value in the diet, which can be attributed tothe BO
seedcontent of beta-carotene, a substance that can be
converted into vitamin A inside the body, which considered
an antioxidant and activator of the immune system
(Prabhakara et al, 2015).
Table 4 referred to the reduction of TBA value through
increasing the concentration of BO seeds in the diet
compared to T1 for the periods after (4, 8, 12) weeks of
the experiment. The lower value of TBA was noted in
T2, T3 and T4 compared to the T1 that recorded the
highest value, where the relationship was inverse between
the amount of the BO seeds and the amount of the TBA.
The obtained results of the present study in agreement
with previous work of Cuong and Chin (2016), who
demonstrated the ability to use BO seed as an antioxidant.
Many studies have reported that the presence of phenolic
compounds in food, prevent certain diseases and cancers
due to their beneficial health properties, which may be
associated with their antioxidant abilities. Furthermore,
Carvalho et al (2006) also stated that a diet that lacks
carotenoids may expose animals to oxidative stress in
tissues in the light of the biochemical functions they play
in living organisms. Additionally, Annatto has been shown
to inhibit the formation of hydroproxide, which leads to
Table 2 : The effect of adding different levels of Bixa orellana (BO)
seeds powder to laying chicken diets on theindex of free
fatty acids (mean ± standard error) for periods 4, 8, 12
weeks.
Free fatty acids
Treatments
4 weeks 8 weeks 12 weeks
T1 0.627 ± 0.009a 0.808 ± 0.025a 0.904 ± 0.032a
T2 0.584 ± 0.029a 0.577 ± 0.006b 0.634 ± 0.076b
T3 0.330 ± 0.021b 0.289 ± c0.045 0.449 ± 0.030c
T4 0.159 ± 0.130c 0.158 ± 0.010d 0.278 ± 0.041d
L.S.D ** ** **
** **: Significant difference (P <0.01),T1 = control treatment
(without addition of annatto seed powder), T2 = add 1% of the
ANATO seed powder, T3, add 2% of the ANATO seed powder, T4
and add 3% of the ANATO seed powder, Different letters within
one column indicate significant differences (P <0.01).
Table 3 : The effect of adding different levels of Bixa orellana (BO)
seeds powder to laying chicken diet on oxidation indices
(peroxide value) / (mean ± standard error) for periods 4,
8, 12 weeks.
Peroxide Value (PV)
Treatments
4 weeks 8 weeks 12 weeks
T1 0.058 ± 0.664a 1.180 ± 0.037a 1.321 ± 0.019a
T2 0.555 ± 0.032ab 0.686 ± 0.015b 1.039 ± 0.046b
T3 0.470 ± 0.035b 0.486 ± 0.020c 0.872 ± 0.018c
T4 0.223 ± 0.019c 0.248 ± 0.041d 0.405 ± 0.016d
L.S.D ** ** **
** **: Significant difference (P <0.01), T1 = control treatment
(without addition of annatto seed powder), T2 = add 1% of the
ANATO seed powder, T3, add 2% of the ANATO seed powder, T4
and add 3% of the ANATO seed powder,Different letters within
one column indicate significant differences (P <0.01).
Table 4 :The effect of adding different levels of Bixa orellana(BO)
seeds powder to the laying chicken diets on the oxidation
index (T.B.A) / (average ± standard error) for periods 4,
8, 12 weeks.
T.B.A
Treatments
4 weeks 8 weeks 12 weeks
T1 0.021 ± 0.331a 0.508 ± 0.005a 0.536 ± 0.018a
T2 0.202 ± 0.003b 0.363 ± 0.022b 0.435 ± 0.029b
T3 0.182 ± 0.005b 0.187 ± 0.025c 0.290 ± 0.033c
T4 0.040 ± 0.004c 0.050 ± 0.001d 0.086 ± 0.032d
L.S.D ** ** **
** **: Significant difference (P <0.01), T1 = add 1% of the ANATO
seed powder, T3, add 2% of the ANATO seed powder, T4 and add
3% of the ANATO seed powder, Different letters within one column
indicate significant differences (P <0.01).
triglyceride oxidation by trapping proxy roots (Haila et
al, 1996). Based on these results, it can be concluded
that Annatto seed powder (BO) revealed antioxidant
activity when added to the diet of laying chickens.
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