Tomato powder in laying hen diets: Effects on concentrations of yolk carotenoids and lipid peroxidation

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1 See discussions, stats, and author profiles for this publication at: Tomato powder in laying hen diets: Effects on concentrations of yolk carotenoids and lipid peroxidation Article in British Poultry Science October 2012 Impact Factor: 0.94 DOI: / Source: PubMed CITATIONS 9 READS authors, including: Fatih Akdemir Inonu University 30 PUBLICATIONS 286 CITATIONS Kazim Sahin Firat University 129 PUBLICATIONS 2,968 CITATIONS SEE PROFILE SEE PROFILE Armagan Hayirli Ataturk University 86 PUBLICATIONS 1,178 CITATIONS SEE PROFILE Available from: Fatih Akdemir Retrieved on: 12 May 2016

2 This article was downloaded by: [Inonu Universitesi] On: 15 October 2012, At: 01:14 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: Registered office: Mortimer House, Mortimer Street, London W1T 3JH, UK British Poultry Science Publication details, including instructions for authors and subscription information: Tomato powder in laying hen diets: effects on concentrations of yolk carotenoids and lipid peroxidation F. Akdemir a, C. Orhan b, N. Sahin b, Dr K. Sahin b & A. Hayirli c a Department of Animal Nutrition, Faculty of Veterinary Medicine, Dicle University, Diyarbakir, Turkey b Department of Animal Nutrition, Faculty of Veterinary Medicine, Firat University, Elazig, Turkey c Department of Animal Nutrition, Faculty of Veterinary Medicine, Atatürk University, Erzurum, Turkey Accepted author version posted online: 24 Sep To cite this article: F. Akdemir, C. Orhan, N. Sahin, Dr K. Sahin & A. Hayirli (2012): Tomato powder in laying hen diets: effects on concentrations of yolk carotenoids and lipid peroxidation, British Poultry Science, DOI: / To link to this article: Disclaimer: This is a version of an unedited manuscript that has been accepted for publication. As a service to authors and researchers we are providing this version of the accepted manuscript (AM). Copyediting, typesetting, and review of the resulting proof will be undertaken on this manuscript before final publication of the Version of Record (VoR). During production and pre-press, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal relate to this version also. PLEASE SCROLL DOWN FOR ARTICLE Full terms and conditions of use: This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. The publisher does not give any warranty express or implied or make any representation that the contents will be complete or accurate or up to date. The accuracy of any instructions, formulae, and drug doses should be independently verified with primary sources. The publisher shall not be liable for any loss, actions, claims, proceedings, demand, or costs or damages whatsoever or howsoever caused arising directly or indirectly in connection with or arising out of the use of this material.

3 Tomato powder in laying hen diets: effects on concentrations of yolk carotenoids and lipid peroxidation F. AKDEMIR, C. ORHAN 1, N. SAHIN 1, K. SAHIN 1 AND A. HAYIRLI 2 Department of Animal Nutrition, Faculty of Veterinary Medicine, Dicle University, Diyarbakir, Turkey, 1 Department of Animal Nutrition, Faculty of Veterinary Medicine, Firat University, Elazig, Turkey and 2 Department of Animal Nutrition, Faculty of Veterinary Medicine, Atatürk University, Erzurum, Turkey Corresponding author: Dr Kazim Sahin, Department of Animal Nutrition, Faculty of Veterinary Science, Firat University, Elazig-Turkey. Phone: / nsahinkm@yahoo.com 1

4 Abstract. 1. The effects of tomato powder supplementation on performance, egg quality, serum and egg yolk carotenoids, vitamins and malondialdehyde (MDA) concentrations in were investigated in laying hens in mid-lay. 2. A total of 90 laying hens, 49 weeks old, were divided into 3 groups consisting of 6 replicate cages, 5 birds per cage. Birds were randomly fed on one of three diets: basal diet and basal diet added with 5 or 10 g tomato powder per kg diet. 3. As tomato powder concentration increased, there were linear increases in feed intake, egg production, egg weight and yolk colour and a linear decrease in feed conversion. Shell weight, shell thickness and Haugh unit remained unchanged in response to dietary treatments. 4. Concentrations of serum and egg yolk lycopene, β-carotene, lutein and vitamin A increased for both diets including tomato powder, whereas MDA decreased linearly with increasing supplemental tomato powder concentration. 5. Tomato powder supplementation increased egg production persistency and increased carotenoids and vitamin A contents in egg yolk, accompanied by reduced yolk lipid peroxidation. INTRODUCTION Carotenoids are pigmented phytochemicals responsible for red, yellow and orange colours of fruits and vegetables (Mangels et al., 1993; Rao and Agarwal, 1999). Tomato, one of the carotenoid-rich fruits, contains large quantities of lycopene (80-90% of total carotenoids), β- carotene (7-10%) and smaller quantities of lutein, α-carotene, γ-carotene, ζ-carotene, violaxanthin, zeaxanthin, neoxanthin, α-cryptoxanthin, β-cryptoxanthin, neurosporene, phytoene, phytofluene, cyclolycopene and some other beneficial molecules such as vitamin E, vitamin C and flavonoids (Bourne and Rice-Evans, 1998; Nguyen and Schwartz, 1999; Khachik et al., 2002; Burns et al., 2003). Lycopene possesses a powerful antioxidant activity, protects cells/tissues from the oxidative damage caused by reactive oxygen species (Di- 2

5 Mascio et al., 1989; Rao and Shen, 2002). In association with these properties, lycopene has been shown to play an important role in preventing certain types of cancer, cardiovascular and degenerative diseases, improving gene regulation, gap-junction communications, immune functions and hormonal and metabolic pathways (Zhang et al., 1991; Gerster, 1993; Meydani et al., 1995; Mayne, 1996; Aust et al., 2003; Rao et al., 2006). Other useful compounds present in tomato such as β-carotene, lutein and flavonoids also exert antioxidant effects (Hertog et al., 1993; Khachik et al., 1995; Mayne, 1996). Acceptability of foods of animal origin is a very important factor in human nutrition (Angelo, 1992; Flachowsky et al., 2002). Some phytochemicals, such as isoflavones, polyphenols and carotenoids, are added to animal diets to improve product quality with respect to colour, tenderness, oxidative stability and storage properties (Hertog et al., 1993; Adlercreutz, 1995; Rock et al., 1996; McCall and Frei, 1999). These beneficial phytochemicals can easily pass into and accumulate in the animal food product (Leeson and Caston, 2004; Cachaldora et al., 2008; Sahin et al., 2008a; Akdemir and Sahin, 2009; Sahin et al., 2010). Therefore, the objective in this study was to determine the effect of dietary tomato powder supplementation on performance, egg quality and lipid peroxidation in mid-lay hens. MATERIALS AND METHODS A total of week-old white laying hens (Lohman LSL, hybrid) were used in accordance with animal welfare regulations at the Umut Tavukculuk of Elazig, Turkey. Hens were assigned randomly to one of three groups. Each treatment consisted of 6 replicate cages of 5 birds per cage. The birds were housed in wire cages and exposed to a 16L:8D illumination cycle. Feed and fresh water were offered ad libitum throughout the experiment. After a 10-d adaptation period, birds were given diets containing 0, 5 or 10 g tomato powder per kg diet for a period of 90 d. Tomato powder (Natural Carotenoid Biomass with Lycopene, Vitan Ltd., Dneprovsky, Ukraine) contained 11% protein and 6% lycopene. 3

6 Experimental diets (Table 1) were stored in black plastic containers at 4 C to avoid photooxidation. Feed consumption was measured weekly and egg production rate and egg weights were recorded daily. At the end of the study, egg quality parameters (egg weight, yolk colour, yolk weight, shell weight, shell thickness and Haugh unit) were measured on two eggs collected randomly from each of 6 replicates per group. Egg yolk colour was determined by using the Roche Color Fan according to the CIE standard colorimetric system. Haugh units were calculated using following formula: Haugh unit = 100 x log (H x W 0.37 ) where H = albumen height, mm and W = egg weight, g (Eisen et al., 1962) after determining albumen height by a micrometer (Saginomiya, TLM-N1010, Japan) and egg weight. Feed samples were analysed in triplicate for crude protein (#988.05), ether extract (#932.06), crude fibre (#962.09), crude ash (#936.07), Ca (#968.08) and P (#965.17) (AOAC, 1990). Energy and amino acid (methionine and lysine) contents were calculated from tabular values listed for the feedstuffs (Jurgens, 1996). A total of 12 eggs from each group were collected randomly and yolks were separated from albumen. For serum analysis, blood samples were collected from the axillary vein of two hens from each of 6 cages per group. Yolk and serum samples were subjected to duplicate analyses for carotenoids (lycopene, β- carotene and lutein) (Stahl and Sies, 1992), vitamins (A and E) (Mori et al., 2003) and MDA (Karatepe, 2004) using high performance liquid chromatography (HPLC, Shimadzu, Kyoto, Japan). All-trans-retinol and α-tocopherol were used as standards (Sigma Chemical Co., St Louis, MO). The equipment for HPLC consisted of a pump (LC-20AD), a Diode Array Detector (SPD-M10A) for carotenoids, vitamins and MDA, a column oven (CTO-10ASVP), an autosampler (SIL-20A) a degasser unit (DGU-20A5), column (Inertsil ODS-3, 250 x 4.6 mm, 5 m) and a computer system with LC solution Software (Shimadzu, Kyoto, Japan). In sample size calculation, 10% improvement in egg lycopene concentration was considered to be significant at type I error of 0.05 with the power of Data were subjected 4

7 to one-way ANOVA using the PROC MIXED procedure of Statistical Analysis System (SAS, 1999). Linear model to test effects of dietary tomato powder supplementation on animal performance and egg quality was y ij = µ + b 0 + L i + e j, where y = response variable, µ = population mean, b 0 = covariate, L = tomato powder supplementation and e = residual error being N (0, 1). Egg production and egg weight data during the adaptation period were used as covariates for corresponding response variables. The model also included orthogonal and polynomial contrasts to determine tomato powder supplementation effects and changes in response variable with increasing dietary tomato powder supplementation. Statistical significance was declared at P < RESULTS The mean egg production (93.33 ± 0.34%) and egg weight (62.10 ± 0.13 g) was similar in all groups during the adaptation period. Hens receiving diets supplemented with tomato powder had greater feed intake, egg production and egg and yolk weights, darker yolk colour and more efficient feed conversion than hens not supplemented with tomato powder (Table 2). As dietary tomato powder concentration increased, feed intake (P < 0.01), egg production (P < 0.001) and egg (P < 0.01) and yolk (P < 0.01) weights and yolk colour (P < ) linearly increased and feed efficiency (P < 0.05) linearly increased (Table 2; Figure). The dietary treatments did not affect shell weight, shell thickness or Haugh units. Serum and egg yolk lycopene and β-carotene concentrations were influenced by tomato powder concentrations (Table 3). Serum and egg yolk lycopene, β-carotene, lutein and vitamin A concentrations increased for hens given diets supplemented with tomato powder (Table 3). The same was true for serum vitamin E concentrations whereas yolk vitamin E concentration did not differ between treatments (Table 3). Hens fed on a diet with tomato powder had lower serum and egg yolk MDA concentrations than hens on a diet without tomato powder. There were linear increases in serum and yolk lycopene, β-carotene, lutein 5

8 and vitamin A concentrations and decreased MDA concentrations in serum and yolk as tomato powder concentration in the diet increased. Despite no changes in yolk vitamin E concentration, its concentration in serum linearly increased with increasing dietary tomato powder concentration. DISCUSSION This experiment tested whether tomato powder supplementation to mid-lay hens improve performance and egg quality in terms of reducing lipid peroxidation through enriching it with vitamins and carotenoids possessing antioxidant properties. Acceptability of foods of animal origin by consumers is influenced by quality factors such as colour, tenderness, oxidative stability and storage properties (Angelo, 1992; Herber and Van Elswyk, 1998; Flachowsky et al., 2002). It is well known that enrichment of poultry diets with carotenoids results in increased concentration in the egg yolk. Pure lycopene or lycopene-rich vegetables such as tomato or tomato by-products have been studied to obtain the darkest egg yolk colour and stable eggs resulting from their increased antioxidant ability against lipid peroxidation (Knoblich et al., 2005; Karadas et al., 2006a; Olson et al., 2008; Sahin et al., 2008a). Lycopene is a predominant carotenoid that gives red colour to tomatoes (Rao and Agarwal, 1999). It is a strong antioxidant and potently reduces oxidative damages of lipids, proteins and DNA (Di-Mascio et al., 1989; Rao and Agarwal, 1999). In agreement with the present data, other studies in which lycopene or lycopene-rich tomato byproducts were supplemented to poultry diets resulted in increased egg yolk colour darkness (Knoblich et al., 2005; Karadas et al., 2006a; Mansoori et al., 2008; Olson et al., 2008). Moreover, in response to tomato powder supplementation, increased egg yolk lycopene concentration was associated with decreased egg yolk MDA concentrations, suggesting that enrichment of diets with vitamins and carotenoids reduced egg yolk lipid peroxidation (Karadas et al., 2006a, b; Olson et al., 2008; Sahin et al., 2008a). It is well documented that lycopene and vitamin E synergistically 6

9 inhibit TBARS formation, another lipid peroxidation indicator (Shixian et al., 2005; Sahin et al., 2006b). Furthermore, Alshatwi et al. (2010) declared that tomato powder was more protective than lycopene against lipid peroxidation in rats. An inverse association between serum lycopene and MDA was also shown in this study and others (Sahin et al., 2006a, 2007, 2008b). Other carotenoids in tomato powder such as lutein and β-carotene also exert biological and antioxidant activities (Khachik et al., 1995; Mayne, 1996). In the present study, lutein and β-carotene concentrations in serum and egg yolk of hens given feed supplemented with tomato powder were significantly higher than control group. Other studies also noted that serum lutein and β-carotene concentrations increased when quails were given feed supplemented with tomato powder (Karadas et al., 2006a, b; Sahin et al. 2007). Tomato powder supplementation to diets increased serum and yolk carotenoids and vitamin A concentrations. Vitamin E concentrations were increased in serum, but not yolk (Table 3). Previous studies investigating tomato powder or pure lycopene supplementation showed increase in both vitamins in serum and egg yolk (Karadas et al., 2005, 2006b; Sahin et al., 2006a, 2006b, 2007, 2008b; Olson et al., 2008). In disagreement with the present study (except for egg weight), Jafari et al. (2006) and Mansoori et al. (2008) achieved the improvements in quality parameters (Haugh unit, shell weight, shell thickness and egg weight) in laying hen supplemented with diets containing dried tomato pomace and tomato pulp. Sahin et al. (2008a), however, reported that egg weight and feed conversion were not affected by lycopene supplementation. In conclusion, carotenoids were effectively transferred from the diet to the egg yolk via tomato powder supplementation to mid-lay hen diets, which resulted in improvements in egg yolk colour and oxidative stability. That is, natural supplements rich in beneficial phytochemicals can be added at up to 10g per kg to poultry diets to improve egg quality. 7

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16 Egg production, % Covariate 1-15 days days days days Time relative to experiment, two weeks days days Figure. Effects of tomato powder (, 0 g/kg;, 5 g/kg;, 10 g/kg) supplementation on egg production. Covariate is average initial egg production of all birds first 10 d prior to the experimental period at the age of 49 weeks. 18

17 Table 1. Ingredient and nutrient composition of the basal diet a Ingredient g/kg Maize Soybean meal Meat-bone meal 50.0 Soy oil 22.7 Limestone 89.0 Dicalcium phosphate 2.5 Vitamin-mineral premix b 6.0 Sodium chloride 2.0 Sodium bicarbonate 2.0 Chemical analyses (g/kg, dry matter basis) c Metabolisable energy, MJ/kg 11,76 Crude protein 170.2±7.5 Crude fat 45.0±2.4 Crude fibre 35.2±1.3 Crude ash 120.8±6.0 Calcium 40.0±1.7 Phosphorus 35.0±1.8 Methionine 4.6 Lysine 9.1 a Tomato powder was added into diets at expense of maize. b Supplied per kg of diet: retinyl acetate, mg; cholecalciferol, 60 µg; dl-α-tocopheryl acetate, 30 mg; menadione sodium bisulphite, 2,5 mg; thiamine-hydrochloride, 3 mg; riboflavin, 7 mg; niacin, 40 mg; d- pantothenic acid, 8 mg; pyridoxine hydrochloride, 4 mg; vitamin B 12, mg; vitamin C, 50 mg; folic acid, 1 mg; D-biotin, mg; choline chloride, 125 mg; Mn (MnSO 4 -H 2 O), 80 mg; Fe (FeSO 4-7H 2 O), 30 mg; Zn (ZnO), 60 mg; Cu (CuSO 4-5H 2 O), 5 mg; Co (CoCl 2-6H 2 O), 0,1 mg; I as KI, 0.4 mg; Se (Na 2 SeO 3 ), 0.15 mg. c Metabolisable energy, methionine and lysine are calculated based on tabular values for feedstuffs (Jurgens, 1996). Others are mean of 3 replicates. 19

18 Table 2. Effects of tomato powder supplementation to hen diets on performance and egg quality (ls-means over a 90-d period) Variable Tomato powder, g/kg Statistical significan ce, P < a SEM S L Q Feed intake, g/d NS Egg production b, % NS Feed consumption c NS Egg weight d, g NS Shell weight, g NS Shell thickness, mm NS NS NS Haugh unit e NS NS NS Egg yolk colour NS Egg yolk weight, g NS a Statistical contrast: S = Tomato powder supplementation effect (hen supplemented with tomato powder vs. hen not supplemented with tomato powder); L = Linear effect of increasing dietary tomato powder; Q = Quadratic effect of increasing dietary tomato powder. NS = not significant. b n = 6 cages, 5 birds per cage. c g feed consumed per g egg mass (egg number x egg weight). d n = 12 eggs per group. e Haugh unit = 100 x log(h x W 0.37 ) where H = albumen height, mm and W = egg weight, g. 20

19 Table 3. Effects of tomato powder supplementation to hen diets on serum-egg yolk carotenoid, vitamin and MDA levels (ls-means over a 90-day period) Variable Tomato powder, g/kg Statistical significan ce, P < a SEM S L Q Serum b, µg/ml Lycopene ND β-carotene Lutein NS Vitamin A NS Vitamin E NS Malondialdehyde NS Egg yolk c, µg/g Lycopene ND β-carotene NS Lutein NS Vitamin A NS Vitamin E NS NS NS Malondialdehyde NS a Statistical contrast: S = Tomato powder supplementation effect (hen supplemented with tomato powder vs. hen not supplemented with tomato powder); L = Linear effect of increasing dietary tomato powder; Q = Quadratic effect of increasing dietary tomato powder. NS = not significant. b n = 12 hens per group. c n = 12 eggs per group. 21

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