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Skin pigmentation in broiler chickens fed various levels of metabolizable energy and xanthophylls from Tagetes erecta
©2012 Poultry Science Association, Inc.
Skin pigmentation in broiler chickens fed various levels of metabolizable energy and xanthophylls from Tagetes erecta J. I. Muñoz-Díaz,* B. Fuente-Martínez,* X. Hernández-Velasco,†1 and E. Ávila-González*
Primary Audience: Nutritionists, Feed Manufacturers, Poultry Producers SUMMARY Two experiments were conducted in broilers raised by sex from 21 to 49 d of age to analyze some factors that affect skin pigmentation. In the first experiment, the effect of feeding broilers 85 ppm of xanthophylls (Xa) from Aztec marigold flowers (Tagetes erecta) and 4 levels of ME (2,800, 3,000, 3,200, and 3,400 kcal/kg) was evaluated. In experiment 2, the objective was to study the effect of feeding 65 ppm of Xa to broilers from 1 to 21 d of age and 4 feeding regimens thereafter: 1) 75 ppm of Xa from 21 to 49 d; 2) 108 ppm of Xa from 35 to 49 d; 3) 141 ppm of Xa from 35 to 49 d; and 4) 162 ppm of Xa from 35 to 49 d. In experiment 1, there was a significant linear effect (P < 0.01) of dietary energy level on the skin pigmentation of live birds (R2 = 68.8%). It was observed that for every 100 kcal/kg increase in ME, skin b+ (yellowness) values increased by 0.13 yellowness units (YU)/d, and by 2 YU at the end of the study. Furthermore, on the basis of consumption time, females gained 1.77 YU for every YU gained by males, and gained 3.77 YU more than the males at the end of the study. In experiment 2, birds fed 75 ppm of Xa from 21 to 49 d had a daily gain of 0.59 YU, and the females had 1.5 YU more than the males at the end of the study. In treatments 2, 3, and 4, a daily reduction of 0.11 YU in skin pigmentation occurred when the birds did not consume dietary pigment from d 21 to 35. This loss was greater in females at 35 d. At the end of the study, the females gained 1.73 YU more than did males. We observed that it is possible to reach more than 18 skin YU values in vivo when feeding higher concentrations of dietary Xa (from Aztec marigold flowers) from 35 to 49 d of age. Key words: Aztec marigold flower, broiler chicken, metabolizable energy, Tagetes erecta, yellow skin pigmentation, xanthophyll 2012 J. Appl. Poult. Res. 21:788–796 http://dx.doi.org/10.3382/japr.2011-00507
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Corresponding author: [email protected]
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*Centro de Enseñanza, Investigación y Extensión en Producción Avícola, Facultad de Medicina Veterinaria y Zootecnia (FMVZ), Universidad Nacional Autónoma de México (UNAM), México Distrito Federal, 13209; and †Departamento de Medicina y Zootecnia de Aves, FMVZ, UNAM, Ciudad Universitaria, México Distrito Federal, 04510
Muñoz-Díaz et al.: SKIN PIGMENTATION IN BROILERS DESCRIPTION OF PROBLEM
Addition of fat in poultry diets as a concentrated source of ME is a common practice. The process of absorption, transport, and deposition of pigments in poultry tissues can be affected by the energy concentration of the diet as well as the type and quality of the fatty acids. Vegetable oils, which are rich in monoglycerides from short-chain fatty acids and long-chain unsaturated fatty acids, are more easily digested and absorbed than animal fats or fats that are saturated, oxidized, or with a high concentration of peroxides [6, 11]. An increase in ME in the diet from the addition of soybean oil resulted in greater broiler skin pigmentation [11]. The objective of the current study was to evaluate the skin b+ (yellowness) values in male and female broilers when dietary ME was increased. A second objective was to understand whether, by increasing the dietary concentration of Xa, it was possible to reduce the time required to achieve the skin yellowness level necessary for commercialization, and to determine whether that response was different between male and female broilers.
MATERIALS AND METHODS The experiments were conducted in accordance with the Guidelines of the Institutional Animal Care and Use Committee of the Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México. Experiment 1 This study was conducted in battery cages within an open-sided house. Ninety-six Ross 308 one-day-old straight-run broiler chickens were obtained from a commercial hatchery [12] and raised on a common starter diet up to 21 d of age. On d 21, the birds were sorted by sex and BW such that each pen had a similar weight and weight distribution. The initial BW at 21 d of age was 650 g on average. Twenty-four males and 24 females were randomly assigned to 4 dietary treatments, with 3 replications of 4 birds each (2 males and 2 females). Treatments consisted of 4 levels of ME (2,800, 3,000, 3,200, and 3,400 kcal of ME/kg). Diets were based on sorghum and soybean meal, and contained
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Food appearance, particularly an intense bright coloration, is a very important characteristic that can determine product preference or rejection by the consumer [1, 2]. In the Mexican poultry industry, consumption of chicken with intense yellow skin and shanks is a deep-rooted cultural characteristic that defines product commercialization [3]. In the initial stages of poultry production, the required yellow coloration was achieved with pigments contained in ingredients used in poultry feeding, such as yellow corn. This ingredient contains a fair amount of carotenoids, which have antioxidant activity and may provide other benefits for human health [4, 5], in addition to their well-known properties of pigmenting broiler skin. However, because of the advances in genetic selection of the fastgrowing strains, the growth period of broiler chickens has been shortened. Thus, to achieve good skin pigmentation, it is necessary to add highly concentrated xanthophyll products to the feed. These products may represent 8 to 10% of the diet cost. Despite this, there are currently more countries where pigmented broiler skin is preferred and with greater intensity [6], which causes more dependency on dietary xanthophyll use for broiler feeding and a higher cost of pigmenting products [2, 7]. A large variety of xanthophylls exist, but not all have the same capacity for pigmenting broiler skin. The main sources of concentrated xanthophylls used for diet formulation are xanthophylls (Xa) from Aztec marigold flowers (Tagetes erecta), the natural source of lutein most widely used; synthetic carotenoids, such as apo-ester and canthaxanthin, and chilies from the Capsicum genre [3, 7]. Reaching adequate skin pigmentation levels depends on several factors: dietary nutritive quality, xanthophyll dosage and consumption time, bird sex, bird health, the feeding program, bird husbandry, and slaughter processing conditions, among others [8–10]. Because of the economic impact of broiler skin pigmentation and because skin yellowness can be affected by several factors, it is important to study and analyze the pigmentation process, emphasizing those factors that influence pigmentation positively and that can be manipulated by the nutritionist.
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Table 1. Composition of the diets for broiler chickens from 21 to 49 d of age in experiment 1 (kg) Item
3,000 kcal of ME/kg
3,200 kcal of ME/kg
3,400 kcal of ME/kg
633.764 232.688 80.995 15.627 14.450 5.330 4.714 3.744 2.709 2.321 1.000 0.835 0.500 0.500 0.173 0.500 0.150 1,000
638.127 243.775 40.414 16.198 14.246 5.330 28.733 3.800 2.727 2.174 1.000 0.835 0.500 0.500 0.141 0.500 0.150 1,000
645.693 254.233 0 16.758 14.048 5.330 51.703 3.856 2.744 2.040 1.000 0.835 0.500 0.500 0.110 0.500 0.150 1,000
600.856 261.631 0 16.886 13.990 5.330 89.134 3.864 2.805 1.893 1.000 0.835 0.500 0.500 0.126 0.500 0.150 1,000
18.00 1.07 0.563 0.86 0.72 0.449 0.9 0.45 0.16
18.00 1.07 0.563 0.86 0.72 0.449 0.9 0.45 0.16
18.00 1.07 0.563 0.86 0.72 0.449 0.9 0.45 0.16
18.00 1.07 0.563 0.86 0.72 0.449 0.9 0.45 0.16
1
Concentration of 15 mg/kg of xanthophylls from Aztec marigold flower (Tagetes erecta). Mineral premix supplied the following per kilogram: manganese, 120 g; zinc, 100 g; iron, 120 g; copper, 10–15 g; iodine, 0.7 g; selenium, 0.4 g; and cobalt, 0.2 g. 3 Vitamin premix supplied the following per kilogram: vitamin A, 20,000,000 IU; vitamin D3, 6,000,000 IU; vitamin E, 75,000 IU; vitamin K3, 9 g; thiamine, 3 g; riboflavin, 8 g; pantothenic acid, 18 g; niacin, 60 g; pyridoxine, 5 g; folic acid, 2 g; biotin, 0.2 g; cyanocobalamin, 16 mg; and ascorbic acid, 200 g. 4 Ethoxyquin. 2
85 ppm of Xa from Aztec marigold flowers (T. erecta; Table 1). Protein and amino acid needs were formulated according to recommendations of the primary breeder [13]. Feed and water were provided ad libitum. At the beginning of the experiment, and every 2 d until 49 d of age, skin b+ values were measured on the right apterial latero-pectoral area by using a reflectance colorimeter [14]. At the end of the study, all birds were slaughtered, processed, and refrigerated for 24 h. Skin yellowness was also measured in refrigerated carcasses on the right side of the breast skin. Data were fitted to a linear regression model in which
X1 represented the dietary ME levels and X2 represented the sex of the bird, as indicator variables (males = 0; females = 1) [15]. Experiment 2 This study was conducted in floor pens within an open-sided house. Three hundred twenty 1-d-old Ross 308 straight-run broiler chickens were raised on a common starter diet containing 65 ppm of Xa through 21 d of age. On d 21, the birds were sorted by sex and BW, such that each pen had a similar weight and weight distribution. The birds were randomly assigned
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Ingredient Sorghum Soybean meal Wheat middlings Calcium phosphate Calcium carbonate Yellow pigment1 Vegetable oil Salt dl-Methionine l-Lysine hydrochloride Choline chloride 60% Mineral premix2 Zinc bacitracin Vitamin premix3 l-Threonine Sodium monensin Antioxidant4 Total Calculated analysis CP, % Lysine, % Methionine, % Methionine + cysteine, % Threonine, % Tryptophan, % Total calcium, % Available phosphorus, % Sodium, %
2,800 kcal of ME/kg
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Table 2. Composition of the diets for broiler chickens from 21 to 49 d of age in experiment 2 (kg) Dietary xanthophylls Item
75 ppm
108 ppm
141 ppm
162 ppm
653.251 255.603 48.298 16.742 14.048 0 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
647.401 255.603 48.298 16.742 14.048 5.0 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
645.201 255.603 48.298 16.742 14.048 7.2 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
643.001 255.603 48.298 16.742 14.048 9.4 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
641.601 255.603 48.298 16.742 14.048 10.8 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
3,176 18.00 1.067 0.567 0.857 0.720 0.449 0.900 0.450 0.160
1
Concentration of 15 mg/kg of xanthophylls from Aztec marigold flower (Tagetes erecta). Vitamin premix supplied the following per kilogram: vitamin A, 20,000,000 IU; vitamin D3, 6,000,000 IU; vitamin E, 75,000 IU; vitamin K3, 9 g; thiamine, 3 g; riboflavin, 8 g; pantothenic acid, 18 g; niacin, 60 g; pyridoxine, 5 g; folic acid, 2 g; biotin, 0.2 g; cyanocobalamin, 16 mg; and ascorbic acid, 200 g. 3 Mineral premix supplied the following per kilogram: manganese, 120 g; zinc, 100 g; iron, 120 g; copper, 10–15 g; iodine, 0.7 g; selenium, 0.4 g; and cobalt, 0.2 g. 4 Ethoxyquin. 2
to 4 dietary treatments consisting of 1) 75 ppm of Xa from 21 to 49 d of age; 2) 108 ppm of Xa from 35 to 49 d of age; 3) 141 ppm of Xa from 35 to 49 d of age; and 4) 162 ppm of Xa from 35 to 49 d of age (Table 2). Treatments contained 4 replications of 20 birds each, with 2 replications (pens) of each sex. Birds in treatments 2 through 4 received a diet without pigment from 21 to 34 d of age. The finisher diets were based on sorghum and soybean meal, and amino acids and protein were formulated to meet the primary breeder recommendations for the bird strain [13]. Feed and water were provided ad libitum. The skin
b+ values were determined in vivo in all birds at 21 d of age, and twice a week afterward through 49 d of age. Body weight gain, feed intake, and FCR were calculated weekly throughout the study. Data for skin yellowness were fitted into a linear regression model in which X1 was the dietary Xa, X2 was the time of pigment consumption (in days), and X3 was the sex (males = 0; females = 1). Growth performance data were analyzed via ANOVA for a completely randomized design [15]. Before the study, it was determined that the minimum sample size to detect significant differences (P < 0.05) in skin b+ val-
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Ingredient Sorghum Soybean meal Vegetable oil Calcium phosphate Calcium carbonate Yellow pigment1 Salt dl-Methionine Vitamin premix2 l-Lysine hydrochloride Choline chloride (60%) Mineral premix3 Zinc bacitracin Sodium monensin Antioxidant4 Total Calculated analysis ME, kcal/ kg CP, % Lysine, % Methionine, % Methionine + cysteine, % Threonine, % Tryptophan, % Total calcium, % Available phosphorus, % Sodium, %
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RESULTS AND DISCUSSION Experiment 1
Figure 1. Effect of increasing dietary ME on skin yellowness of broiler chickens fed 75 ppm of xanthophylls from Aztec marigold flowers (Tagetes erecta) from 21 to 49 d of age.
pigmentation than males at the end of the study (49 d of age; P < 0.01) in a magnitude of 3.77 YU. This occurred regardless of the dietary ME level (Table 3). The linear equation explaining this relationship was Y = −46.55 ± 7.67 + 0.02 ± 0.002 (ME level) + 3.77 ± 1.12 (sex) (R2 = 68.80, P < 0.01; Figure 1). Regarding the length of time that Xa were consumed, it was observed that females gained 1.77 YU more for every YU gained by the males at 49 d of age (Table 4). Moreover, it was observed that for every 100 kcal/kg of dietary ME, skin pigmentation increased by 0.13 YU/d. This relationship is explained by the following equation: Y = −41.62 ± 2.06 +0.13 ± 0.001 (ME level) + 1.77 ± 0.294 (sex) + 0.594 ± 0.016 (time of pigment consumption, in days). This finding clearly shows a difference in skin pigmentation attributable to sex. It is well
Table 3. Skin b+ (yellowness) values of male and female broiler chickens in vivo and in 24-h refrigerated carcasses after feeding 85 ppm of xanthophylls from Aztec Marigold flowers (Tagetes erecta) and increasing levels of ME In vivo Item ME, kcal/kg 2,800 3,000 3,200 3,400 Average Linear effect Equation
Females (n = 24)
Males (n = 24)
14.38 ± 0.69 10.45 ± 1.55 18.51 ± 2.11 13.46 ± 1.33 21.70 ± 1.38 17.63 ± 2.88 26.69 ± 1.65 23.40 ± 1.69 20.32 16.23 R2 = 68.80, P < 0.01 Y = −44.91 ± 8.65 + 0.02 ± 0.002 (ME)
24-h refrigerated carcass Average 12.41 15.98 19.66 25.04
Females (n = 24)
Males (n = 24)
Average
33.21 ± 2.98 32.44 ± 2.34 31.54 32.74 ± 1.49 31.77 ± 3.06 32.25 40.88 ± 2.06 40.52 ± 1.25 40.70 47.59 ± 1.72 51.17 ± 1.94 49.38 38.60 38.97 R2 = 65.31, P < 0.01 Y = −52.74 ± 11.14 + 0.029 ± 0.003 (ME)
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Skin pigmentation at 21 d of age was not significantly different between treatments (P < 0.01). Feed consumption and, consequently, pigment consumption were not affected by ME concentration of the diet (P > 0.05). A significant linear relationship was detected in both live birds (R2 = 68.8%) and refrigerated carcasses (R2 = 65.31%) between skin pigmentation and dietary ME level. For every 100 kcal of ME/kg increase in the diet, skin pigmentation increased by 2 skin b+ values (yellowness units; YU) in males and females at the end of the study. Pigmentation results in live birds observed in the current study differ from those reported by Alpízar [16], in which feeding ME levels of 2,990, 3,030, and 3,150 kcal/kg had no effect on skin pigmentation. This could be due to the small range of ME levels used in that study. In contrast, in the current study, the range of ME levels between treatments was wider (200 kcal/ kg). Such an increase in skin pigmentation was probably due to the liposolubility of xanthophylls, which could facilitate their absorption, transportation, and deposition in tissues when ME was increased by adding soybean oil [17, 18]. When sex was used for statistical evaluation, it was observed that females had greater skin
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Table 4. Gradual increase in skin yellowness (b+) of male and female broiler chickens fed 85 ppm of xanthophylls from Aztec Marigold flowers (Tagetes erecta) and increasing dietary ME levels (2,800 to 3,400 kg/kg) from 21 to 49 d of age 2,800 Item
Females
3,000 Males
Females
3,200 Males
Females
3,400 Males
Females
Males
Table 5. Growth performance of broiler chickens fed different levels of xanthophylls from Aztec Marigold flowers (Tagetes erecta) from 21 to 49 d of age1 Treatment
BW gain,3 kg/bird
Feed consumption, kg/bird
FCR
Pigment consumption, g/bird
75 ppm 108 ppm 141 ppm 162 ppm SEM
2.164ª 2.187ª 2.172ª 2.362ª 0.050
4.456a 4.384a 4.459a 4.331a 0.160
2.059ª 2.005ª 2.053ª 1.834ª 0.102
0.334ab 0.294b 0.399ab 0.438a 0.030
2
a,b
Means with different superscripts within a column differ significantly (P < 0.05). n for each mean = 80. 2 Xanthophylls were fed from 21 to 49 d of age for treatment 1, and from 35 to 49 d of age for treatments 2 through 4. 3 Initial BW of 591 g at 21 d of age. 1
Table 6. Luminosity (L+), redness (a+), and yellowness (b+) in the skin of live broiler chickens fed 75 ppm of yellow xanthophylls from Aztec Marigold flowers (Tagetes erecta) from 21 to 49 d of age L+ Item Treatment 75 ppm SEM Equation
Male
a+ Female
Male
b+ Female
Male
64.12a 2.11a 3.06ª 22.37a 0.22 0.15 0.49 Y = 7.2 ± 0.63 + 0.59 ± 0.03 (time of pigment consumption, in days) + 1.50 ± 0.060 (sex) 67.84ª
Female 25.08a
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Days of pigment consumption 0 1.48 1.30 1.64 1.25 1.45 1.29 1.74 1.10 3 1.58 1.84 3.50 4.16 3.98 5.49 5.00 6.31 5 3.63 2.21 4.26 4.16 5.55 5.49 6.81 6.31 7 4.34 2.91 4.57 5.22 6.22 7.28 8.35 8.97 10 6.77 5.61 7.89 6.84 9.62 11.09 11.70 12.30 12 7.99 5.86 9.21 7.71 12.28 12.39 14.92 13.89 14 8.84 4.69 10.49 7.00 15.11 15.12 18.92 15.07 17 8.52 6.06 11.76 7.37 18.19 14.41 18.31 17.69 19 10.65 7.60 13.72 7.04 15.48 15.50 22.66 19.97 21 11.60 6.15 14.05 6.19 17.90 17.30 23.61 19.09 24 12.26 7.47 14.06 7.47 18.89 19.48 24.91 21.74 26 12.61 12.23 15.40 11.20 19.98 18.47 23.30 23.94 28 14.38 12.37 17.18 13.46 21.70 17.59 26.69 22.83 SEM 0.48 0.50 0.45 0.68 0.84 0.78 0.94 1.01 Linear effect R2 = 74.67, P < 0.01 Equation Y = −41.62 ± 2.06 + 0.013 ± 0.001 (ME level) + 1.77 ± 0.294 (sex) + 0.594 ± 0.016 (time of pigment consumption, in days)
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Figure 3. Skin yellowness of male broiler chickens fed various concentrations of dietary xanthophylls from Aztec marigold flowers (Tagetes erecta). *Control.
Figure 4. Skin yellowness of female broiler chickens fed various concentrations of dietary xanthophylls from Aztec marigold flowers (Tagetes erecta). *Control.
Figure 2. Effect of dietary pigment withdrawal on skin yellowness of male and female broiler chickens from 21 to 35 d of age.
known in the field that females have a greater pigmentation capacity. It is also known that skin-pigmenting xanthophylls are deposited mostly in the body fat and that the ability to deposit body fat is a highly heritable trait, which varies depending on bird strain and sex [19, 20]. However, there are no reports showing that this difference is maintained when different ME levels from added soybean oil are fed. Furthermore, there are no studies in which such a difference in skin pigmentation has been quantified. Observations from this study could be used to reach the expected pigmentation level for each sex more accurately and efficiently, considering that strain and sex must be evaluated for their capacity to reach a determined degree of pigmentation. Experiment 2
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No significant differences were observed between treatments (P > 0.05) for BW gain,
feed intake, or FCR. Pigment consumption was greater in birds fed 162 ppm of Xa compared with birds fed 108 ppm of Xa. No differences were detected in overall pigment consumption between birds fed 75 ppm from 21 to 49 d of age and birds fed 141 ppm from 35 to 49 d of age (Table 5). Skin yellowness in live birds fed 75 ppm of Xa from 21 to 49 d of age showed a daily increase of 0.59 YU [Y = 7.2 ± 0.63 + 0.59 ± 0.03(time of pigment consumption, in days) + 1.50 ± 0.060 (sex)]. However, the females had 1.5 YU more than the males (Table 6). When birds did not receive dietary Xa from 21 to 35 d of age, there was a reduction in skin pigmentation of 0.11 YU for every day that these birds did not consume Xa. The loss in skin pigmentation was greater for females (1.46 YU; P < 0.05) [Y = 5.85 ± 1.05 to 0.11 ± 0.042 (time of pigment consumption, in days) + 1.46 ± 0.32 (sex)] (R2 = 70.0; Figure 2). When dietary Xa were fed at high dosages (141 and 162 ppm) only from 35 to 49 d of age, skin yellowness was not significantly different from that observed in birds fed 75 ppm of Xa from 21 to 49 d of age (P > 0.05; Tables 6 and 7). No significant difference was observed between males and females fed 141 or 162 ppm from 35 to 49 d of age compared with males or females fed 75 ppm from 21 to 49 d of age (Figures 3 and 4). Martínez et al. [21] found that when adding 80 ppm of Xa for 3 to 4 wk before slaughter, the degree of skin pigmentation was adequate to meet the market standards for Mexico. In the present study, it was demonstrated that it is
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Table 7. Luminosity (L+), redness (a+), and yellowness (b+) in the skin of broiler chickens fed 108, 141, or 162 ppm of yellow xanthophylls from Aztec Marigold flowers (Tagetes erecta) from 35 to 49 d of age L+ Item Treatment 108 ppm 141 ppm 162 ppm SEM Equation
a+
b+
Male
Female
Male
Female
Male
Female
65.72a 65.84a 63.55a
64.53a 63.45a 64.47a
2.59a 1.62a 3.36a
3.82a 3.05a 3.92a
18.68b 21.98ab 20.39ab
19.60b 21.22ab 23.24ab
0.22 0.15 0.49 Y = 1.21 ± 1.54 + 0.026 ± 0.01 (ppm of treatment) + 1.14 ± 0.060 (time of pigment consumption, in days) + 1.73 ± 0.45 (sex), R2 = 70.0
Means with different superscripts within a column differ significantly (P < 0.05).
possible to shorten the time needed to reach the required skin pigmentation level by increasing the dietary concentration of the pigment. Furthermore, the time frame in which pigmentation could be affected by external factors was able to be reduced. Moreover, increasing the dietary pigment level could also be used as a corrective measure to improve low levels of skin pigmentation. In a similar fashion as experiment 1, females had greater skin pigmentation than males, in a magnitude of 1.73 YU. In the study reported herein, the skin pigmentation loss was also evaluated from 21 to 35 d of age. To our knowledge, measurement of the skin pigmentation loss when birds do not receive dietary pigment after being fed dietary pigment regularly has not been reported previously. The greater skin pigmentation of female chickens compared with males had not been reported previously either.
CONCLUSIONS AND APPLICATIONS According to the results of the experiments reported herein,
1. Skin pigmentation can be increased by increasing dietary ME. For every 100 kcal/kg, skin pigmentation increased by 2 YU at 49 d of age. 2. Regardless of the dietary energy level, females had greater skin pigmentation than males at 49 d of age. Females had consistently greater skin pigmentation than males throughout the period of pigment consumption (21 to 49 d of age). 3. It is possible to meet the market expectations for skin pigmentation in Mexico by
feeding 141 ppm of Xa from 35 to 49 d of age.
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796 15. Kuehl, R. 2001. Diseño de experimentos: Principios estadísticos de diseño y análisis de investigación. 2nd ed. Universidad de Arizona, Tucson, AZ. 16. Alpízar, S. O. 1993. Respuesta de los parámetros productivos de pollos de engorda a diferentes niveles de energía metabolizable. Rev. Vet. Mex. 24:211–215. 17. Tyczkowski, J. K., and P. B. Hamilton. 1986. Absorption, transport and deposition in chickens of lutein diester, a carotenoid extracted from Marigold (Tagetes erecta). Poult. Sci. 65:1526–1531. 18. Raghavan, V. 2001. Pigmentation in broilers. Feed Mix 9:14–15. 19. Wiseman, J. 1988. Nutrition and carcass fat. Poult. Int. 27:12–14.
20. Leenstra, F. R., and R. Pit. 1988. Fat depositions in a broiler sire strain. Poult. Sci. 67:1–9. 21. Martínez, P. M., C. A. Cortes, and G. E. Ávila. 2004. Evaluación de tres niveles de pigmento de flor de cempasúchil (Tagetes erecta) sobre la pigmentación de la piel en pollos de engorda. Tec. Pecu. Mex. 42:105–111.
Acknowledgments
This project was sponsored by Programa de Apoyo a Proyectos de Investigación e Innovación Tecnológica (PAPIIT) [Dirección General de Apoyo al Personal Académico (DGAPA); Project IN203910-3], Universidad Nacional Autónoma de México. Downloaded from http://japr.oxfordjournals.org/ at University of North Dakota on June 2, 2015
Skin pigmentation in broiler chickens fed various levels of metabolizable energy and xanthophylls from Tagetes erecta J. I. Muñoz-Díaz,* B. Fuente-Martínez,* X. Hernández-Velasco,†1 and E. Ávila-González*
Primary Audience: Nutritionists, Feed Manufacturers, Poultry Producers SUMMARY Two experiments were conducted in broilers raised by sex from 21 to 49 d of age to analyze some factors that affect skin pigmentation. In the first experiment, the effect of feeding broilers 85 ppm of xanthophylls (Xa) from Aztec marigold flowers (Tagetes erecta) and 4 levels of ME (2,800, 3,000, 3,200, and 3,400 kcal/kg) was evaluated. In experiment 2, the objective was to study the effect of feeding 65 ppm of Xa to broilers from 1 to 21 d of age and 4 feeding regimens thereafter: 1) 75 ppm of Xa from 21 to 49 d; 2) 108 ppm of Xa from 35 to 49 d; 3) 141 ppm of Xa from 35 to 49 d; and 4) 162 ppm of Xa from 35 to 49 d. In experiment 1, there was a significant linear effect (P < 0.01) of dietary energy level on the skin pigmentation of live birds (R2 = 68.8%). It was observed that for every 100 kcal/kg increase in ME, skin b+ (yellowness) values increased by 0.13 yellowness units (YU)/d, and by 2 YU at the end of the study. Furthermore, on the basis of consumption time, females gained 1.77 YU for every YU gained by males, and gained 3.77 YU more than the males at the end of the study. In experiment 2, birds fed 75 ppm of Xa from 21 to 49 d had a daily gain of 0.59 YU, and the females had 1.5 YU more than the males at the end of the study. In treatments 2, 3, and 4, a daily reduction of 0.11 YU in skin pigmentation occurred when the birds did not consume dietary pigment from d 21 to 35. This loss was greater in females at 35 d. At the end of the study, the females gained 1.73 YU more than did males. We observed that it is possible to reach more than 18 skin YU values in vivo when feeding higher concentrations of dietary Xa (from Aztec marigold flowers) from 35 to 49 d of age. Key words: Aztec marigold flower, broiler chicken, metabolizable energy, Tagetes erecta, yellow skin pigmentation, xanthophyll 2012 J. Appl. Poult. Res. 21:788–796 http://dx.doi.org/10.3382/japr.2011-00507
1
Corresponding author: [email protected]
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*Centro de Enseñanza, Investigación y Extensión en Producción Avícola, Facultad de Medicina Veterinaria y Zootecnia (FMVZ), Universidad Nacional Autónoma de México (UNAM), México Distrito Federal, 13209; and †Departamento de Medicina y Zootecnia de Aves, FMVZ, UNAM, Ciudad Universitaria, México Distrito Federal, 04510
Muñoz-Díaz et al.: SKIN PIGMENTATION IN BROILERS DESCRIPTION OF PROBLEM
Addition of fat in poultry diets as a concentrated source of ME is a common practice. The process of absorption, transport, and deposition of pigments in poultry tissues can be affected by the energy concentration of the diet as well as the type and quality of the fatty acids. Vegetable oils, which are rich in monoglycerides from short-chain fatty acids and long-chain unsaturated fatty acids, are more easily digested and absorbed than animal fats or fats that are saturated, oxidized, or with a high concentration of peroxides [6, 11]. An increase in ME in the diet from the addition of soybean oil resulted in greater broiler skin pigmentation [11]. The objective of the current study was to evaluate the skin b+ (yellowness) values in male and female broilers when dietary ME was increased. A second objective was to understand whether, by increasing the dietary concentration of Xa, it was possible to reduce the time required to achieve the skin yellowness level necessary for commercialization, and to determine whether that response was different between male and female broilers.
MATERIALS AND METHODS The experiments were conducted in accordance with the Guidelines of the Institutional Animal Care and Use Committee of the Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México. Experiment 1 This study was conducted in battery cages within an open-sided house. Ninety-six Ross 308 one-day-old straight-run broiler chickens were obtained from a commercial hatchery [12] and raised on a common starter diet up to 21 d of age. On d 21, the birds were sorted by sex and BW such that each pen had a similar weight and weight distribution. The initial BW at 21 d of age was 650 g on average. Twenty-four males and 24 females were randomly assigned to 4 dietary treatments, with 3 replications of 4 birds each (2 males and 2 females). Treatments consisted of 4 levels of ME (2,800, 3,000, 3,200, and 3,400 kcal of ME/kg). Diets were based on sorghum and soybean meal, and contained
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Food appearance, particularly an intense bright coloration, is a very important characteristic that can determine product preference or rejection by the consumer [1, 2]. In the Mexican poultry industry, consumption of chicken with intense yellow skin and shanks is a deep-rooted cultural characteristic that defines product commercialization [3]. In the initial stages of poultry production, the required yellow coloration was achieved with pigments contained in ingredients used in poultry feeding, such as yellow corn. This ingredient contains a fair amount of carotenoids, which have antioxidant activity and may provide other benefits for human health [4, 5], in addition to their well-known properties of pigmenting broiler skin. However, because of the advances in genetic selection of the fastgrowing strains, the growth period of broiler chickens has been shortened. Thus, to achieve good skin pigmentation, it is necessary to add highly concentrated xanthophyll products to the feed. These products may represent 8 to 10% of the diet cost. Despite this, there are currently more countries where pigmented broiler skin is preferred and with greater intensity [6], which causes more dependency on dietary xanthophyll use for broiler feeding and a higher cost of pigmenting products [2, 7]. A large variety of xanthophylls exist, but not all have the same capacity for pigmenting broiler skin. The main sources of concentrated xanthophylls used for diet formulation are xanthophylls (Xa) from Aztec marigold flowers (Tagetes erecta), the natural source of lutein most widely used; synthetic carotenoids, such as apo-ester and canthaxanthin, and chilies from the Capsicum genre [3, 7]. Reaching adequate skin pigmentation levels depends on several factors: dietary nutritive quality, xanthophyll dosage and consumption time, bird sex, bird health, the feeding program, bird husbandry, and slaughter processing conditions, among others [8–10]. Because of the economic impact of broiler skin pigmentation and because skin yellowness can be affected by several factors, it is important to study and analyze the pigmentation process, emphasizing those factors that influence pigmentation positively and that can be manipulated by the nutritionist.
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Table 1. Composition of the diets for broiler chickens from 21 to 49 d of age in experiment 1 (kg) Item
3,000 kcal of ME/kg
3,200 kcal of ME/kg
3,400 kcal of ME/kg
633.764 232.688 80.995 15.627 14.450 5.330 4.714 3.744 2.709 2.321 1.000 0.835 0.500 0.500 0.173 0.500 0.150 1,000
638.127 243.775 40.414 16.198 14.246 5.330 28.733 3.800 2.727 2.174 1.000 0.835 0.500 0.500 0.141 0.500 0.150 1,000
645.693 254.233 0 16.758 14.048 5.330 51.703 3.856 2.744 2.040 1.000 0.835 0.500 0.500 0.110 0.500 0.150 1,000
600.856 261.631 0 16.886 13.990 5.330 89.134 3.864 2.805 1.893 1.000 0.835 0.500 0.500 0.126 0.500 0.150 1,000
18.00 1.07 0.563 0.86 0.72 0.449 0.9 0.45 0.16
18.00 1.07 0.563 0.86 0.72 0.449 0.9 0.45 0.16
18.00 1.07 0.563 0.86 0.72 0.449 0.9 0.45 0.16
18.00 1.07 0.563 0.86 0.72 0.449 0.9 0.45 0.16
1
Concentration of 15 mg/kg of xanthophylls from Aztec marigold flower (Tagetes erecta). Mineral premix supplied the following per kilogram: manganese, 120 g; zinc, 100 g; iron, 120 g; copper, 10–15 g; iodine, 0.7 g; selenium, 0.4 g; and cobalt, 0.2 g. 3 Vitamin premix supplied the following per kilogram: vitamin A, 20,000,000 IU; vitamin D3, 6,000,000 IU; vitamin E, 75,000 IU; vitamin K3, 9 g; thiamine, 3 g; riboflavin, 8 g; pantothenic acid, 18 g; niacin, 60 g; pyridoxine, 5 g; folic acid, 2 g; biotin, 0.2 g; cyanocobalamin, 16 mg; and ascorbic acid, 200 g. 4 Ethoxyquin. 2
85 ppm of Xa from Aztec marigold flowers (T. erecta; Table 1). Protein and amino acid needs were formulated according to recommendations of the primary breeder [13]. Feed and water were provided ad libitum. At the beginning of the experiment, and every 2 d until 49 d of age, skin b+ values were measured on the right apterial latero-pectoral area by using a reflectance colorimeter [14]. At the end of the study, all birds were slaughtered, processed, and refrigerated for 24 h. Skin yellowness was also measured in refrigerated carcasses on the right side of the breast skin. Data were fitted to a linear regression model in which
X1 represented the dietary ME levels and X2 represented the sex of the bird, as indicator variables (males = 0; females = 1) [15]. Experiment 2 This study was conducted in floor pens within an open-sided house. Three hundred twenty 1-d-old Ross 308 straight-run broiler chickens were raised on a common starter diet containing 65 ppm of Xa through 21 d of age. On d 21, the birds were sorted by sex and BW, such that each pen had a similar weight and weight distribution. The birds were randomly assigned
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Ingredient Sorghum Soybean meal Wheat middlings Calcium phosphate Calcium carbonate Yellow pigment1 Vegetable oil Salt dl-Methionine l-Lysine hydrochloride Choline chloride 60% Mineral premix2 Zinc bacitracin Vitamin premix3 l-Threonine Sodium monensin Antioxidant4 Total Calculated analysis CP, % Lysine, % Methionine, % Methionine + cysteine, % Threonine, % Tryptophan, % Total calcium, % Available phosphorus, % Sodium, %
2,800 kcal of ME/kg
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Table 2. Composition of the diets for broiler chickens from 21 to 49 d of age in experiment 2 (kg) Dietary xanthophylls Item
75 ppm
108 ppm
141 ppm
162 ppm
653.251 255.603 48.298 16.742 14.048 0 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
647.401 255.603 48.298 16.742 14.048 5.0 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
645.201 255.603 48.298 16.742 14.048 7.2 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
643.001 255.603 48.298 16.742 14.048 9.4 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
641.601 255.603 48.298 16.742 14.048 10.8 3.855 2.726 0.500 1.992 1.000 0.835 0.500 0.500 0.150 1,000
3,176 18.00 1.067 0.567 0.857 0.720 0.449 0.900 0.450 0.160
1
Concentration of 15 mg/kg of xanthophylls from Aztec marigold flower (Tagetes erecta). Vitamin premix supplied the following per kilogram: vitamin A, 20,000,000 IU; vitamin D3, 6,000,000 IU; vitamin E, 75,000 IU; vitamin K3, 9 g; thiamine, 3 g; riboflavin, 8 g; pantothenic acid, 18 g; niacin, 60 g; pyridoxine, 5 g; folic acid, 2 g; biotin, 0.2 g; cyanocobalamin, 16 mg; and ascorbic acid, 200 g. 3 Mineral premix supplied the following per kilogram: manganese, 120 g; zinc, 100 g; iron, 120 g; copper, 10–15 g; iodine, 0.7 g; selenium, 0.4 g; and cobalt, 0.2 g. 4 Ethoxyquin. 2
to 4 dietary treatments consisting of 1) 75 ppm of Xa from 21 to 49 d of age; 2) 108 ppm of Xa from 35 to 49 d of age; 3) 141 ppm of Xa from 35 to 49 d of age; and 4) 162 ppm of Xa from 35 to 49 d of age (Table 2). Treatments contained 4 replications of 20 birds each, with 2 replications (pens) of each sex. Birds in treatments 2 through 4 received a diet without pigment from 21 to 34 d of age. The finisher diets were based on sorghum and soybean meal, and amino acids and protein were formulated to meet the primary breeder recommendations for the bird strain [13]. Feed and water were provided ad libitum. The skin
b+ values were determined in vivo in all birds at 21 d of age, and twice a week afterward through 49 d of age. Body weight gain, feed intake, and FCR were calculated weekly throughout the study. Data for skin yellowness were fitted into a linear regression model in which X1 was the dietary Xa, X2 was the time of pigment consumption (in days), and X3 was the sex (males = 0; females = 1). Growth performance data were analyzed via ANOVA for a completely randomized design [15]. Before the study, it was determined that the minimum sample size to detect significant differences (P < 0.05) in skin b+ val-
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Ingredient Sorghum Soybean meal Vegetable oil Calcium phosphate Calcium carbonate Yellow pigment1 Salt dl-Methionine Vitamin premix2 l-Lysine hydrochloride Choline chloride (60%) Mineral premix3 Zinc bacitracin Sodium monensin Antioxidant4 Total Calculated analysis ME, kcal/ kg CP, % Lysine, % Methionine, % Methionine + cysteine, % Threonine, % Tryptophan, % Total calcium, % Available phosphorus, % Sodium, %
0 ppm
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RESULTS AND DISCUSSION Experiment 1
Figure 1. Effect of increasing dietary ME on skin yellowness of broiler chickens fed 75 ppm of xanthophylls from Aztec marigold flowers (Tagetes erecta) from 21 to 49 d of age.
pigmentation than males at the end of the study (49 d of age; P < 0.01) in a magnitude of 3.77 YU. This occurred regardless of the dietary ME level (Table 3). The linear equation explaining this relationship was Y = −46.55 ± 7.67 + 0.02 ± 0.002 (ME level) + 3.77 ± 1.12 (sex) (R2 = 68.80, P < 0.01; Figure 1). Regarding the length of time that Xa were consumed, it was observed that females gained 1.77 YU more for every YU gained by the males at 49 d of age (Table 4). Moreover, it was observed that for every 100 kcal/kg of dietary ME, skin pigmentation increased by 0.13 YU/d. This relationship is explained by the following equation: Y = −41.62 ± 2.06 +0.13 ± 0.001 (ME level) + 1.77 ± 0.294 (sex) + 0.594 ± 0.016 (time of pigment consumption, in days). This finding clearly shows a difference in skin pigmentation attributable to sex. It is well
Table 3. Skin b+ (yellowness) values of male and female broiler chickens in vivo and in 24-h refrigerated carcasses after feeding 85 ppm of xanthophylls from Aztec Marigold flowers (Tagetes erecta) and increasing levels of ME In vivo Item ME, kcal/kg 2,800 3,000 3,200 3,400 Average Linear effect Equation
Females (n = 24)
Males (n = 24)
14.38 ± 0.69 10.45 ± 1.55 18.51 ± 2.11 13.46 ± 1.33 21.70 ± 1.38 17.63 ± 2.88 26.69 ± 1.65 23.40 ± 1.69 20.32 16.23 R2 = 68.80, P < 0.01 Y = −44.91 ± 8.65 + 0.02 ± 0.002 (ME)
24-h refrigerated carcass Average 12.41 15.98 19.66 25.04
Females (n = 24)
Males (n = 24)
Average
33.21 ± 2.98 32.44 ± 2.34 31.54 32.74 ± 1.49 31.77 ± 3.06 32.25 40.88 ± 2.06 40.52 ± 1.25 40.70 47.59 ± 1.72 51.17 ± 1.94 49.38 38.60 38.97 R2 = 65.31, P < 0.01 Y = −52.74 ± 11.14 + 0.029 ± 0.003 (ME)
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Skin pigmentation at 21 d of age was not significantly different between treatments (P < 0.01). Feed consumption and, consequently, pigment consumption were not affected by ME concentration of the diet (P > 0.05). A significant linear relationship was detected in both live birds (R2 = 68.8%) and refrigerated carcasses (R2 = 65.31%) between skin pigmentation and dietary ME level. For every 100 kcal of ME/kg increase in the diet, skin pigmentation increased by 2 skin b+ values (yellowness units; YU) in males and females at the end of the study. Pigmentation results in live birds observed in the current study differ from those reported by Alpízar [16], in which feeding ME levels of 2,990, 3,030, and 3,150 kcal/kg had no effect on skin pigmentation. This could be due to the small range of ME levels used in that study. In contrast, in the current study, the range of ME levels between treatments was wider (200 kcal/ kg). Such an increase in skin pigmentation was probably due to the liposolubility of xanthophylls, which could facilitate their absorption, transportation, and deposition in tissues when ME was increased by adding soybean oil [17, 18]. When sex was used for statistical evaluation, it was observed that females had greater skin
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Table 4. Gradual increase in skin yellowness (b+) of male and female broiler chickens fed 85 ppm of xanthophylls from Aztec Marigold flowers (Tagetes erecta) and increasing dietary ME levels (2,800 to 3,400 kg/kg) from 21 to 49 d of age 2,800 Item
Females
3,000 Males
Females
3,200 Males
Females
3,400 Males
Females
Males
Table 5. Growth performance of broiler chickens fed different levels of xanthophylls from Aztec Marigold flowers (Tagetes erecta) from 21 to 49 d of age1 Treatment
BW gain,3 kg/bird
Feed consumption, kg/bird
FCR
Pigment consumption, g/bird
75 ppm 108 ppm 141 ppm 162 ppm SEM
2.164ª 2.187ª 2.172ª 2.362ª 0.050
4.456a 4.384a 4.459a 4.331a 0.160
2.059ª 2.005ª 2.053ª 1.834ª 0.102
0.334ab 0.294b 0.399ab 0.438a 0.030
2
a,b
Means with different superscripts within a column differ significantly (P < 0.05). n for each mean = 80. 2 Xanthophylls were fed from 21 to 49 d of age for treatment 1, and from 35 to 49 d of age for treatments 2 through 4. 3 Initial BW of 591 g at 21 d of age. 1
Table 6. Luminosity (L+), redness (a+), and yellowness (b+) in the skin of live broiler chickens fed 75 ppm of yellow xanthophylls from Aztec Marigold flowers (Tagetes erecta) from 21 to 49 d of age L+ Item Treatment 75 ppm SEM Equation
Male
a+ Female
Male
b+ Female
Male
64.12a 2.11a 3.06ª 22.37a 0.22 0.15 0.49 Y = 7.2 ± 0.63 + 0.59 ± 0.03 (time of pigment consumption, in days) + 1.50 ± 0.060 (sex) 67.84ª
Female 25.08a
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Days of pigment consumption 0 1.48 1.30 1.64 1.25 1.45 1.29 1.74 1.10 3 1.58 1.84 3.50 4.16 3.98 5.49 5.00 6.31 5 3.63 2.21 4.26 4.16 5.55 5.49 6.81 6.31 7 4.34 2.91 4.57 5.22 6.22 7.28 8.35 8.97 10 6.77 5.61 7.89 6.84 9.62 11.09 11.70 12.30 12 7.99 5.86 9.21 7.71 12.28 12.39 14.92 13.89 14 8.84 4.69 10.49 7.00 15.11 15.12 18.92 15.07 17 8.52 6.06 11.76 7.37 18.19 14.41 18.31 17.69 19 10.65 7.60 13.72 7.04 15.48 15.50 22.66 19.97 21 11.60 6.15 14.05 6.19 17.90 17.30 23.61 19.09 24 12.26 7.47 14.06 7.47 18.89 19.48 24.91 21.74 26 12.61 12.23 15.40 11.20 19.98 18.47 23.30 23.94 28 14.38 12.37 17.18 13.46 21.70 17.59 26.69 22.83 SEM 0.48 0.50 0.45 0.68 0.84 0.78 0.94 1.01 Linear effect R2 = 74.67, P < 0.01 Equation Y = −41.62 ± 2.06 + 0.013 ± 0.001 (ME level) + 1.77 ± 0.294 (sex) + 0.594 ± 0.016 (time of pigment consumption, in days)
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Figure 3. Skin yellowness of male broiler chickens fed various concentrations of dietary xanthophylls from Aztec marigold flowers (Tagetes erecta). *Control.
Figure 4. Skin yellowness of female broiler chickens fed various concentrations of dietary xanthophylls from Aztec marigold flowers (Tagetes erecta). *Control.
Figure 2. Effect of dietary pigment withdrawal on skin yellowness of male and female broiler chickens from 21 to 35 d of age.
known in the field that females have a greater pigmentation capacity. It is also known that skin-pigmenting xanthophylls are deposited mostly in the body fat and that the ability to deposit body fat is a highly heritable trait, which varies depending on bird strain and sex [19, 20]. However, there are no reports showing that this difference is maintained when different ME levels from added soybean oil are fed. Furthermore, there are no studies in which such a difference in skin pigmentation has been quantified. Observations from this study could be used to reach the expected pigmentation level for each sex more accurately and efficiently, considering that strain and sex must be evaluated for their capacity to reach a determined degree of pigmentation. Experiment 2
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No significant differences were observed between treatments (P > 0.05) for BW gain,
feed intake, or FCR. Pigment consumption was greater in birds fed 162 ppm of Xa compared with birds fed 108 ppm of Xa. No differences were detected in overall pigment consumption between birds fed 75 ppm from 21 to 49 d of age and birds fed 141 ppm from 35 to 49 d of age (Table 5). Skin yellowness in live birds fed 75 ppm of Xa from 21 to 49 d of age showed a daily increase of 0.59 YU [Y = 7.2 ± 0.63 + 0.59 ± 0.03(time of pigment consumption, in days) + 1.50 ± 0.060 (sex)]. However, the females had 1.5 YU more than the males (Table 6). When birds did not receive dietary Xa from 21 to 35 d of age, there was a reduction in skin pigmentation of 0.11 YU for every day that these birds did not consume Xa. The loss in skin pigmentation was greater for females (1.46 YU; P < 0.05) [Y = 5.85 ± 1.05 to 0.11 ± 0.042 (time of pigment consumption, in days) + 1.46 ± 0.32 (sex)] (R2 = 70.0; Figure 2). When dietary Xa were fed at high dosages (141 and 162 ppm) only from 35 to 49 d of age, skin yellowness was not significantly different from that observed in birds fed 75 ppm of Xa from 21 to 49 d of age (P > 0.05; Tables 6 and 7). No significant difference was observed between males and females fed 141 or 162 ppm from 35 to 49 d of age compared with males or females fed 75 ppm from 21 to 49 d of age (Figures 3 and 4). Martínez et al. [21] found that when adding 80 ppm of Xa for 3 to 4 wk before slaughter, the degree of skin pigmentation was adequate to meet the market standards for Mexico. In the present study, it was demonstrated that it is
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Table 7. Luminosity (L+), redness (a+), and yellowness (b+) in the skin of broiler chickens fed 108, 141, or 162 ppm of yellow xanthophylls from Aztec Marigold flowers (Tagetes erecta) from 35 to 49 d of age L+ Item Treatment 108 ppm 141 ppm 162 ppm SEM Equation
a+
b+
Male
Female
Male
Female
Male
Female
65.72a 65.84a 63.55a
64.53a 63.45a 64.47a
2.59a 1.62a 3.36a
3.82a 3.05a 3.92a
18.68b 21.98ab 20.39ab
19.60b 21.22ab 23.24ab
0.22 0.15 0.49 Y = 1.21 ± 1.54 + 0.026 ± 0.01 (ppm of treatment) + 1.14 ± 0.060 (time of pigment consumption, in days) + 1.73 ± 0.45 (sex), R2 = 70.0
Means with different superscripts within a column differ significantly (P < 0.05).
possible to shorten the time needed to reach the required skin pigmentation level by increasing the dietary concentration of the pigment. Furthermore, the time frame in which pigmentation could be affected by external factors was able to be reduced. Moreover, increasing the dietary pigment level could also be used as a corrective measure to improve low levels of skin pigmentation. In a similar fashion as experiment 1, females had greater skin pigmentation than males, in a magnitude of 1.73 YU. In the study reported herein, the skin pigmentation loss was also evaluated from 21 to 35 d of age. To our knowledge, measurement of the skin pigmentation loss when birds do not receive dietary pigment after being fed dietary pigment regularly has not been reported previously. The greater skin pigmentation of female chickens compared with males had not been reported previously either.
CONCLUSIONS AND APPLICATIONS According to the results of the experiments reported herein,
1. Skin pigmentation can be increased by increasing dietary ME. For every 100 kcal/kg, skin pigmentation increased by 2 YU at 49 d of age. 2. Regardless of the dietary energy level, females had greater skin pigmentation than males at 49 d of age. Females had consistently greater skin pigmentation than males throughout the period of pigment consumption (21 to 49 d of age). 3. It is possible to meet the market expectations for skin pigmentation in Mexico by
feeding 141 ppm of Xa from 35 to 49 d of age.
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Acknowledgments
This project was sponsored by Programa de Apoyo a Proyectos de Investigación e Innovación Tecnológica (PAPIIT) [Dirección General de Apoyo al Personal Académico (DGAPA); Project IN203910-3], Universidad Nacional Autónoma de México. Downloaded from http://japr.oxfordjournals.org/ at University of North Dakota on June 2, 2015