Shahsavari

   Influences of Different Sources of Natural Pigments on the 
   Color and Quality of Eggs from Hens Fed a Wheat‐Based Diet 
Research Article K. Shahsavari1* 
 
1
 Department of Animal Science, Shabestar Branch, Islamic Azad University, Shabestar, Iran 
 
 
Received on: 21 Mar 2014 
Revised on: 19 May 2014 
Accepted on: 31 May 2014 
Online Published on: Mar 2015 
 

 
 
*Correspondence E‐mail: [email protected] 
© 2010 Copyright by Islamic Azad University, Rasht Branch, Rasht, Iran 
Online version is available on: www.ijas.ir

This experiment was conducted to evaluate effects of different sources of natural pigments on yolk color
and laying hen performance. A completely randomized design with six treatments and six replicates was
used.  The six experimental treatments were: a control diet containing yellow corn grain, a reference diet
containing wheat and barley grain instead of corn; a red pepper diet composed of reference diet and 2% red
pepper; a dried carrot diet containing the reference diet and 5% dried carrot meal; a dried tomato pulp diet
composed of the reference diet and 5% dried tomato pulp and an alfalfa meal diet containing the reference
  diet and 5% alfalfa meal. Results indicated that egg weight, daily egg mass, feed conversion ratio and egg
 
yolk color were not affected by treatments. In conclusion, if the goal of producers is an acceptable level of
 
egg yolk color and performance is not under consideration, then the use of a 5% alfalfa meal in a wheat-
  barley based diet is suggested.

 
KEY WORDS  carotenoid, egg quality, layer, pigment, yolk color. 

yolk color. Although corn is used at high levels in the diet

INTRODUCTION of laying hens, it does not enhance the red pigmentation of
The degree of yolk color is an important criterion in table the yolk properly (Gurbuz et al. 2003). In countries like
eggs for consumption as well as manufacturing of egg- Iran, where the production of yellow corn is limited, wheat
containing food products (De-Groote, 1970). Therefore, and barley are the only grains usually considered by poultry
there has been interest in evaluating the yolk-pigmenting producers in diet formulations. Wheat-barley based diets
properties of different feed sources. The color of egg yolks usually contain insufficient color pigments to enhance egg
is produced by oxycarotenoids, as xanthophyll pigments, yolk pigmentation (Gurbuz et al. 2003). Therefore, when
derived from the feed ingredients (Zahroojian et al. 2011). wheat and barley are used as the main sources of energy,
Sources of xanthophylls can be natural or synthetic. The additional pigmenting additives should be added. Recently,
supplementation of the laying hen's diet with synthetic poultry rations have been in a continuing state of change
pigments is more expensive than natural coloring sources, with many new ingredients and special oxycarotenoid con-
since the former must be imported (Hoppe, 1998). The centrates being developed. These products include red pep-
principal natural feed sources of xanthophylls used during per, dried tomato pulp, and dried carrot meal. There is lim-
the past several decades include yellow corn and alfalfa ited information available regarding the effectiveness of
meal. Corn is one of the essential feedstuffs of the hen's different natural pigmenting sources (red pepper, dried to-
diet, and it contains lutein pigment, which affects the egg mato pulp, alfalfa meal and dried carrot meal) in wheat-

Iranian Journal of Applied Animal Science (2015) 5(1), 167-172 167

 
 Influence of Natural Pigmenting Sources on Laying Hens

barley based diets of layers. Thus, the objective of the pre- Egg production (%), egg weight (g), egg mass
sent study was to evaluate the effects of different natural (g/day/bird) and feed conversion ratio were recorded daily
pigment sources in diets on yolk color and laying hen per- (Jafari et al. 2006). Egg quality characteristics were meas-
formance. ured for five eggs from each replicate that were collected
randomly during the 4th, 7th and 12th week of the experi-
MATERIALS AND METHODS mental period. Egg width and length were measured by
using slide calipers (Ogawa Seiki Co. Ltd; Tokyo, Japan).
This study used a completely randomized design with 540 The egg was then carefully broken on a glass plate to meas-
Hy-Line (w-36) hens at 40 weeks of age. They were se- ure egg quality characteristics. Egg shape index was calcu-
lected based on egg production and body weight from 7000 lated according to Hasin et al. (2006) for each egg from its
birds, and were allocated to 6 treatments with 6 replicates, average length and width. The egg shape index= (average
each of 15 hens. width of egg/average length of egg) × 100. The albumen
The experiment continued for a period of 12 weeks (40 to index was calculated by dividing the average height of thick
52 weeks of age). In the pre-experimental period (36 to 40 albumen by its width according to Hasin et al. (2006). For
wk), egg production, egg weight, and initial body weight each egg, the shell thickness was determined at three differ-
were recorded for hens fed on a typical corn-soybean meal ent points in the middle part of the egg using a Mitutoyo
diet. The hens were selected based on the pre-experimental caliper 7313 micrometer with 0.01 mm precision. Haugh
egg production and body weight so that the average per- units were calculated from the weight and height of albu-
formance of the hens in each treatment was similar at the men of egg using the formula suggested by Haugh (1937).
start of the experiment. Each cage was provided with an
individual feeder and two automatic drinkers. The cages HU= 100 log (H+7.57-1.7 W0.37
were located in a temperature-controlled room, and the pho-
toperiod during the experiment was fixed at 16 hrs per day. Where:
Red pepper, alfalfa meal, dried tomato pulp and dried HU: haugh unit.
carrot meal used in the experiment were analyzed for dry H: height of thick albumen.
matter (DM), metabolizable energy (ME), crude protein W: egg weight (g).
(CP), crude fat, crude fiber and ash. The analyzed composi-
tion of the natural pigment sources is presented in Table 1. The yolk color score was determined by comparing with
The DM contents of experimental natural pigment sources the roche yolk color (RYC) fan (F. Hoffmann-La-Roche
were determined by drying at 102 ˚C for 16 h in a forced air Ltd., Basal, Switzerland). The RYC fan is a standardized
oven. tool which shows the range of yolk colors from 1 (very
The ME contents of the sources were calculated by using light yellow) to 15 (very dark yellow) as produced under
the formula suggested by Wiseman (1987). The crude pro- natural feeding conditions (Vulleumier, 1969). The yolk
tein, crude fat, crude fiber and ash contents of the sources index was calculated as the ratio of average yolk height to
were measured according to methods 976.06, 920.39, average yolk width following removal of the yolk from the
987.10 and 942.05 of AOAC (1990), respectively. Calcium albumen. Also, egg shell weight and specific gravity (Den-
and phosphorus contents were determined by the methods sitometer, Mettler-Toledo, Iso-14001, Switzerland) were
of Page et al. (1982). measured according to Holder and Bradford (1979). Egg
Diets were formulated using an UFFDA feed formulation yolk cholesterol content was measured in accordance with
package according to the nutrient requirements of white standards of the Association of Official Analytical Chem-
laying hens (NRC, 1994). They were balanced to be iso- ists (AOAC, 1990). Data were analyzed as a completely
nitrogenous and iso-caloric, and to meet all other nutrient randomized design. Statistical analysis of data was carried
requirements of the birds. Red pepper, alfalfa meal, dried out using the SAS statistical program (SAS, 2002), and
tomato pulp and dried carrot meal were analyzed in order to differences between means were tested with Duncan’s mul-
permit appropriate feed formulation. The composition and tiple range tests at P < 0.05.
calculated analyses of experimental diets is outlined in Ta-
ble 2.
All treatments (except the control diet) were supple- RESULTS AND DISCUSSION
mented with a commercial cellulase and xylanase enzyme The data on mean production performance (egg production,
mixture. Diets were made in mash form and provided daily egg weight, egg mass and feed conversion ratio) of the lay-
according to expected intake, and water was available ad ing hens from 40 to 52 weeks of age are presented in Table
libitum throughout the experiment. 3.

168  Iranian Journal of Applied Animal Science (2015) 5(1), 167-172

Shahsavari

Table 1 Analyzed composition of natural pigment sources

DM ME Crude protein Crude fat Crude fiber Ash Calcium Phosphorus
Pigmented product
(%) (kcal/kg) (%) (%) (%) (%) (%) (%)
Red pepper 88.21 1450 14.04 9.01 23.20 10.53 0.60 0.37
Dried tomato pulp 94.46 1760 20.40 8.80 33.67 4.50 0.52 0.47
Alfalfa meal 92.22 1673 19.84 3.42 21.08 10.60 1.47 0.23
Dried carrot meal 93.86 1721 4.03 0.52 11.63 5.30 0.66 0.50
DM: dry matter and ME: metabolizable energy.
DM and ME estimated to according to Wiseman (1987).

Table 2 Composition and calculated analyses of experimental diets

Dietary treatments
Control diet Reference diet Red pepper Dried tomato pulp Alfalfa meal Dried carrot meal
Ingredients, %
Yellow corn 59.27 0.00 0.00 0.00 0.00 0.00
Soybean meal 22.38 18.06 17.50 15.50 15.46 15.71
Barley grain 0.00 28.53 27.19 26.10 26.21 25.93
Wheat grain 0.00 35.00 35.00 35.00 35.00 35.00
Soybean oil 5.67 7.10 7.10 7.10 7.10 7.10
Dicalcium phosphate 1.65 1.10 1.10 1.10 1.10 1.10
DL-methionine 0.19 0.17 0.17 0.18 0.16 0.17
Lysine 0.00 0.06 0.04 0.12 0.09 0.09
Oyster shell 9.92 9.10 9.00 9.00 8.98 9.01
Sodium chloride 0.42 0.38 0.35 0.35 0.35 0.34
Vitamin premix1 0.25 0.25 0.25 0.25 0.25 0.25
Mineral premix2 0.25 0.25 0.25 0.25 0.25 0.25
Enzyme mixture3 0.00 0.05 0.05 0.05 0.05 0.05
Red pepper 0.00 0.00 2.00 0.00 0.00 0.00
Dried tomato pulp 0.00 0.00 0.00 5.00 0.00 0.00
Alfalfa meal 0.00 0.00 0.00 0.00 5.00 0.00
Dried carrot meal 0.00 0.00 0.00 0.00 0.00 5.00
Total 100.00 100.00 100.00 100.00 100.00 100.00
Calculated content:
AME, kcal/kg 2895 2898 2896 2896 2897 2898
Crude protein, % 14.89 15.06 15.10 14.92 14.97 15.01
Calcium, % 3.84 3.85 3.83 3.84 3.84 3.85
Available P, % 0.31 0.32 0.32 0.31 0.30 0.30
Methionine, % 0.35 0.36 0.36 0.35 0.35 0.36
Met + Cys, % 0.58 0.60 0.59 0.58 0.60 0.59
Lysine, % 0.71 0.73 0.73 0.72 0.72 0.73
1
Vitamin premix supplied per kg of diet: vitamin A: 9000 IU; vitamin B 12 : 0.015 mg; vitamin D: 2000 IU; vitamin E: 18 IU; vitamin K 3 : 2 mg; vitamin B 1 : 1.78 mg; vitamin
B 2 : 6.6 mg; vitamin B 6 : 3 mg; Niacin: 30 mg; Pantothenic acid: 10 mg; Biotin: 0.15 mg and Choline: 1500 mg.
2
Mineral premix supplied per kg of diet: Cu: 10 mg; I: 0.99 mg; Fe: 50 mg; Mn: 100 mg; Se: 0.08 mg and Zn: 100 mg.
3
Enzyme mixture contained a commercial cellulase and xylanase enzymes. 

Table 3 Effects of dietary treatments on performance of laying hens from 40 to 52 weeks of age
Egg production Egg weight Egg mass Feed conversion ratio
Treatments
(%) (g) (g/day/bird) (g feed/g egg mass)
Control diet 75.84 64.77b 49.20c 1.962ab
ab
Reference diet 79.28 66.98 53.04a 1.854c
a
Red pepper 77.65 68.00 52.78ab 1.890bc
Dried carrot meal 77.20 66.04ab 50.99abc 1.936abc
Alfalfa meal 75.75 66.41ab 50.24bc 1.992a
ab
Dried tomato pulp 79.89 65.64 52.44ab 1.896bc
SEM 0.669 0.382 0.441 0.015
The means within the same column with at least one common letter, do not have significant difference (P>0.05).
SEM: standard error of the means.

There were no significant differences in egg production Also, a significant (P<0.05) increase in egg mass was ob-
(P>0.05) between the dietary treatments. A significant served for hens on the reference diet, the 2% red pepper
(P<0.05) increase in egg weight was observed from hens diet and the 5% dried tomato pulp diet compared to the
receiving the red pepper diet compared to the control diet. control diet.

169  Iranian Journal of Applied Animal Science (2015) 5(1), 167-172

 Influence of Natural Pigmenting Sources on Laying Hens

The highest and lowest egg mass were observed for the There were no significant differences (P>0.05) in egg
reference diet and control diet treatments, respectively. The shell thickness, egg shell weight, shape index, yolk index,
best feed conversion ratio was observed on the reference specific gravity, haugh units and cholesterol content in yolk
diet. Overall, the reference diet, the 2% red pepper diet and among treatments. In a study by Calislar and Uygur (2010),
the 5% dried tomato pulp diet gave better performance than dry tomato pulp had a significant effect on the egg shape
the other treatments. index and egg yolk index, whereas, dry tomato pulp had no
The results of egg quality characteristics (Table 4) significant effect on the albumen index and haugh units. In
showed that there were no significant differences between the current study, egg shell thickness was not affected by
treatments (P>0.05) in egg thickness, egg shell weight, dietary treatments. This result could be due to the nearly
shape index, yolk index, specific gravity, haugh units and similar calcium, phosphorus and vitamin D 3 contents in all
cholesterol content in yolks. Egg yolk colors of all treat- diets. In agreement with our results, Jafari et al. (2006) re-
ments differed significantly (P<0.05) from each other. The ported no significant differences in egg shell thickness and
highest and lowest scores of yolk color were obtained for haugh units of laying hens fed on diets containing up to 50,
eggs from hens receiving the diets containing red pepper 100 and 150 kg/t dried tomato pulp compared to hens fed
(14.33) and the reference diet (1.58), respectively. on a control diet. In contrast, Gregoriades et al. (1984)
The treatments imposed in the present study produced no found that the inclusion of dried tomato pulp in layer diets
significant effects (P>0.05) on egg production of layer improved the shell thickness.
hens. This result can be attributed to the similar energy and Egg yolk colors of eggs from hens fed all experimental
crude protein contents of all dietary treatments. Pigment diets differed significantly (P<0.05) from each other. The
supplementation has not been associated with changes in highest and lowest score of yolk color was obtained with
production in previous studies (Angeles and Scheideler, the diet containing red pepper (14.33) and with the refer-
1998; Garcia et al. 2002; Soto-Salanova, 2003). This non- ence diet (1.58), respectively. Red pepper contains capsan-
significant difference agreed well with the results of previ- thin and capsorubin leading to red color in yolk (Fletcher
ous reports by Dotas et al. (1999), Santos-Bocanegra et al. and Halloran, 1981; Karunajeewa, 1980). Thus, when red
(2004) and Skider et al. (1998) who worked with tomato pepper was used as a color additive to the reference diet at
pulp, red pepper and carrot meal, respectively. The results 2%, reddish or red egg yolk colors scored higher than 14,
regarding egg production suggest that the use of 2% red scores that are preferred in the cake industry (Fletcher and
pepper, 5% dried carrot pulp, 5% alfalfa meal or 5% dried Halloran, 1981; Papa et al. 1985). The roche yolk color
tomato pulp in the diet of laying hens has no detrimental (RYC) value of the reference diet containing 28.53% barley
effect on egg production. The egg weight from the control and 35% wheat and no color additive was found quite low
diet was significantly lower (P<0.05) than that from the red (1.58). All natural pigment sources added resulted in yolk
pepper treatment. color scores higher than that provided by the control group.
In agreement with our results, Yannakopoulos et al. Skider et al. (1998) found a 1.87 yolk color score from a
(1992) found that tomato meal resulted in greater egg 62% wheat-based diet after 3 weeks. Wheat-based diets
weight and suggested that this could be a consequence of its usually fail to produce eggs with a standard yolk color on
high lysine content. On the other hand, Mitsuhiro et al. the RYC fan score (Saha et al. 1999). Although wheat is the
(1994) did not find any improvement of egg weight when main crop produced in Iran, it cannot be used as the main
dietary red pepper was used. Egg mass was significantly ingredient of laying hen diets due to its adverse effect on
different (P<0.05) among treatments. A significant egg yolk color. However, the addition of 5% dried tomato
(P<0.05) increase in egg mass was observed with the refer- pulp or 5% dried carrot meal into that diet resulted in RYC
ence diet, 2% red pepper diet, and the 5% dried tomato pulp values of 3 and 3.92, respectively, which are low, but
diet compared to the control diet treatment. The highest and higher than for the control group. Skider et al. (1998) re-
lowest egg mass were from the reference diet and control ported a yolk color value of 3.12 during the 3rd week from
treatment, respectively. Feed conversion ratios for hens on 4% dried carrot meal supplementation. This value was very
the reference diet, red pepper diet and dried tomato pulp close to the value obtained in our experiment. The addition
diet were better than other treatments (P<0.05). The lowest of 5% alfalfa meal into the reference diet resulted in a RYC
and highest feed conversion ratios were from the reference value of 6.08, which is close to the optimum RYC value
diet and the 5% alfalfa meal diet. The increase in feed con- preferred by Iranian consumers. As expected, the diet con-
version ratio in the alfalfa meal treatment may be attributed taining corn resulted in a darker egg yolk color (RYC=5.75)
to factors such as higher levels of crude fiber, protease in- than the diet containing wheat and barley (RYC=1.58).
hibitors and anti-nutritional factors (such as tannin) (Ali et Yellow corn is among the conventional sources of carote-
al. 2003). noids and rich in zeaxanthin.

170  Iranian Journal of Applied Animal Science (2015) 5(1), 167-172

Shahsavari

Table 4 Effects of dietary treatments on quality characteristics of eggs from layer hens during 40 to 52 weeks of age
Experimental diets
Variables
Control diet Reference diet Red pepper Dried carrot meal Alfalfa meal Dried tomato pulp SEM
Shell thickness (mm) 0.35 0.34 0.35 0.33 0.34 0.36 0.453
Egg shell weight (g) 5.13 5.14 5.17 5.16 5.15 5.15 0.100
Shape index (%) 73.8 74.4 75.7 75.5 76.3 74.6 0.004
Albumen index (%) 10.32 9.89 11.01 10.46 9.97 10.09 0.401
Yolk index (%) 0.40 0.40 0.40 0.39 0.41 0.39 0.002
Yolk color score 5.75c 1.58f 14.33a 3.92d 6.08b 3.00e 1.001
Specific gravity 1.081 1.080 1.081 1.078 1.081 1.081 0.001
Haugh unit 96.16 95.63 98.73 98.19 95.86 94.26 0.850
Cholesterol (mg/g) 6.11 6.75 6.20 6.01 6.32 6.16 0.069
The means within the same column with at least one common letter, do not have significant difference (P>0.05).
SEM: standard error of the means. 

Saxena et al. (1982) reported that feeding hens diets con- pepper, dried carrot meal and dried tomato pulp in wheat-
taining 400 g/kg of yellow corn increased markedly the egg barley based diets of laying hens in order to produce table
yolk score of eggs from 2 to 8 in 3-7 days. The results indi- eggs with yolk colors that are desirable for consumers
cated that the addition of red pepper and alfalfa meal to the without altering laying performance.
reference diet instead of carrot pulp and tomato pulp can be
used to increase dietary xanthophylls levels and enhance ACKNOWLEDGEMENT
yolk color. Jafari et al. (2006) reported that tomato by-
products had no effect on the egg yolk color in laying hens. The author is thankful to “Shabestar Branch, Islamic Azad
However, Dotas et al. (1999) and Yannakopoulos et al. University” for their financial support of present project.
(1992) found that yolk color was higher in eggs produced
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