During storage the percent level of palmitic acid

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estru kwasu β-apo-8-karotenowego (grupa IV). W 42 dniu życia, po 4 reprezentatywne dla. 217 każdej grupy kogutki i kurki dekapitowano, preparowano mięśnie ...
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Annals of Animal Science, 2008, 8, 167-174

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Effect of dietary supplementation of vitamin E, antioxidants and a synthetic carotenoid

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on changes chicken breast meat quality during storage

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National Research Institute of Animal Production, Department of Animal Nutrition and Feed

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Science, 32-083 Balice, Poland

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J. Koreleski 1, S. Świątkiewicz

Study was conducted as a part of NRIAP statutory activity, project no. 5211.1

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Abstract

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Cobb chickens were allocated to 4 groups and from 22 to 42 days of age fed diets with 4 %

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rapeseed oil and 1 % of liquid fish fat (stabilized ethoxyquin, 250 mg/kg) containing in 1 kg

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15.9 mg vitamin E (group I-control) or supplemented with 150 mg dl-α-tocopheryl acetate

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(group II), 48.6 mg BHA+EQ+BHT antioxidant mixture (group III) and 40 mg · kg -1 ethyl

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ester of β-apo-8-carotenoic acid (group IV). At 42 day of age 4 males and 4 females from

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each group were killed, breast muscles were excised, divided to two parts (left and right) and

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frozen (-20oC). One part was analyzed for fatty acids composition of lipid fraction and for

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thiobarbituric acid reactive substances (TBARS) and vitamin E content. Boiled meat was

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evaluated in sensory test. The second part of muscle was kept frozen (-20oC) during 6 month

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and then chemical and sensory analyses were made in the same manner. Fatty acids

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Corresponding author e-mail: [email protected]

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composition was analysed by gas chromatography, α-tocopherol by HPLC. TBARS content

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was expressed in mg of malonylaldehyde/kg of meat.

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After long frozen storage the reduced level of saturated fatty acids (SFA) was observed in

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meat lipids whereas level of n-3 polyunsaturated fatty acids (PUFA) and PUFA/SFA ratio

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was enlarged. Advantageously was also changed PUFA n-6/n-3 ratio (7.7 vs 5.5). Vitamin E

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and TBARS content in long stored meat was similar to those present in short stored meat. In

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boiled long stored meat as compare to short stored meat a significant worsening of flavour

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was observed in all groups and taste was worsened when synthetic antioxidants and yellow

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carotenoid was added to diet. At the dietary level 15.9 mg kg -1 of natural α-tocopherol from

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feed components the similar quality of stored air proof packed breast meat was stated as when

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α-tocopheryl acetate was added. Natural vitamin E content seems to be sufficient.

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Key words: storage changes, broiler breast meat, synthetic antioxidant, synthetic carotenoid,

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vitamin E, nutrition

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Deterioration of meat quality during storage is caused in great part by oxidative changes

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of lipid fractions (Gray et al., 1996) especially rich in polyunsaturated fatty acids and addition

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of antioxidants to the diet could counteract to these undesirable effect (Morrisey et al., 1997).

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Positive effect of vitamin E or synthetic antioxidants of phenolic and non-phenolic structure

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or β-carotene and zeaxanthine enriched diets on susceptability to peroxidation was reported in

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muscle (Bartov and Bornstein, 1977 and 1981; Maraschiello et al., 1999; Bailey et al., 1996;

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Woodall et al., 1996) and in other tissues. The more rapidly absorbed from digestive tract are

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probably these natural or synthetic phenolic antioxidants, e.g. vitamin E or butylated

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hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) - which are more soluble in fat

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(Lin et al., 1989) than non-phenolic ethoxyquin (EQ).

Introduction

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The objective of this study was to evaluate the effect of dietary α-tocopheryl acetate, a

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mixture of BHA, BHT and EQ or synthetic yellow carotenoid added to diet on changes in

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fatty acid composition of lipid fraction, TBARS and vitamin E content and sensory properties

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of boiled meat during long frozen storage of chicken breast meat.

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Material and methods

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The broiler trial was conducted with 160 sexed Cobb chickens from 22 to 42 days of age.

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Before the experiment chickens were fed with standard starter diet. At 22 days of age

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chickens were allocated to 4 groups and fed basal diet (Table 1) not supplemented with

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vitamin E and contained 4 % rapeseed oil and 1 % of stabilized liquid fish fat (ethoxyquin,

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250 mg/kg). Control group was fed the basal diet containing in 1 kg 15.9 mg vitamin E (group

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I-control) and experimental groups were fed basal diet supplemented with 150 mg · kg-1 of dl-

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α-tocopheryl acetate, BASF (group II), and group III with 48.6 mg · kg-1 of BHA + EQ +

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BHT antioxidant mixture, Franklin (in it: butylated hydroxytoluene 28.6 mg, ethoxyquin 14.3

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mg, butylated hydroxyanisol 5.72 mg) or 40 mg · kg-1 of ethyl ester of β-apo-8-carotenoic

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acid, BASF (group IV).

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At the end of experiment 32 representative chickens were chosen (4 males and 4 females

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from each group) and killed. Breast muscles (left and right) were excised, packed to air proof

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plastic bags and frozen (-20oC). During two weeks the left parts of meat (short stored) were

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analyzed for fatty acids composition of lipid fraction, TBARS and vitamin E content and

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evaluated in sensory test. The right part was kept frozen (-20oC) during 6 month (long stored)

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and then analyzed in the same manner.

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The fatty acids (FA) composition of breast meat lipids was determined with a GC Varian

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3400 gas chromatograph equipped with a CP-Wax 58, 25 m x 0.53 mm, 1.0 μm column and

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expressed as % of total FA. Samples were extracted as described by Folch et al. (1957) and

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evaporated under nitrogen, saponified with NaOH, converted to methyl esters (Morrison and

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Smith, 1964), extracted with hexane and separated. Peaks areas measured with Star

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Chromatography Workstation software (Varian Star 4.5). As a measure of oxidative stability

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in meat thiobarbituric acid reactive substances (TBARS) were determined according to Salih

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et al. (1987) with modifications of Pikul et al. (1989). Values were expressed as mg

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malondialdehyde · kg-1. The content of α-tocopherol was determined with HPLC Merck-

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Hitachi equipped with LiChroCART 250-4 Superspher 100 RP-18, 4 μm column and FL, Ex.

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295 nm, Em. 350 nm detector - according to Manz and Phillip (1981). Sensory analysis of

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meat was made before and after freeze storage by panel of six persons. Panelists tested boiled

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meat and ranked flavour, taste, juiciness and tenderness (Baryłko-Pikielna, 1975) in 4-point

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scale (from 2 - not accepted; to 5 – very good).

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For each group separately the effect of frozen storage was statistically evaluated. Data

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were subjected to one-way factorial analysis of variance (Statistica ver. 5.0 PL) and

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differences were examined by Duncan’s multiple range test.

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Results During long frozen storage the percent level of palmitic acid (C16:0) and linolic acid (C18:2)

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decreased but stearic acid (C18:0), arachidonic acid (C20:4), eicosapentaenoic acid (C20:5,

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EPA) and docosaheksaenoic acid (C22:6,

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groups (Table 2). The increase of oleic acid (C18:1) and decrease of linolenic acid (C18:3, n-3 )

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was found only in control chickens. In total, after long storage period the level of saturated

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fatty acids (SFA) was reduced in lipids, whereas level of n-3 polyunsaturated fatty acids

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(PUFA) and PUFA/SFA ratio was enlarged (Table 3). Advantageously was also changed

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PUFA n-6/n-3 ratio. These effects were not related to kind of supplement added to

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experimental diet.

n-3,

n-3,

DHA) levels increased in meat lipids of every

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Vitamin E and TBARS content in long stored meat was similar to short stored meat (Table

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4). A slight, statistically not confirmed tendency to lover vitamin E and greater TBARS

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content in meat was only observed. In long stored meat after boiling as compare to short

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stored a significant (P≤0.05) worsening of flavour was observed in all groups. Long stored

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meat from chickens fed diet with synthetic antioxidants and yellow carotenoid had also worse

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taste than short stored one.

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Discussion Observed differences in fatty acid profiles of breast lipids in short cold stored meat

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and after 6 month of cold storage are probably a result of changes in meat fat content,

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direction of oxidation and degree of fatty acid saturation or desaturation. It may take place

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perhaps when meat was stored in air proof bags, made unfreeze and prepared to the analysis.

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The above changes refer to fatty acid profile in meat fat, but not to the real content of fatty

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acids in meat.

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Reduction of linolenic and linolic acids in chicken meat and partly palmitic acid as

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effect of frozen storage was reported (Miteva and Bakalivanova, 2006), however Coetzee and

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Hoffman (2001) did not find any changes of fatty acids proportions over time of meat storage

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from chickens fed diets with high doses of 120-200 mg vitamin E. In frozen pig ham stearic

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acid level was higher and palmitic acid lower after 30 days of storage (Kingsley et al., 1978).

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In our earlier experiment (Koreleski and Świątkiewicz, 2006) the contribution of palmitic acid

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level was however decreased, whereas level of PUFA n-3 elevated in fat of frozen stored

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breast meat from chickens fed diet with rapeseed oil.

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Vitamin E and TBARS content in long stored meat was similar to short stored meat. A

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slight, statistically not confirmed tendency to lover vitamin E and higher TBARS content in

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long stored meat was only observed.

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The positive effect of vitamin E or synthetic antioxidants of phenolic and non-phenolic

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structure or β-carotene and zeaxanthine enriched diets on susceptibility to peroxidation was

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reported in muscle (Bartov and Bornstein, 1977, 1981; Maraschiello et al., 1999; Bailey et al.

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1996; Woodall et al., 1996) and in other tissues. As synthetic antioxidants are normally not

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deposited to the meat they may prohibit oxidation in diets.

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In boiled long stored meat as compare to short stored a significant worsening of flavour

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was observed (P≤0.05) in all groups. Stored meat from chickens fed diet with synthetic

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antioxidants and yellow carotenoid when boiled had also worse taste than short stored meat.

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It is concluded that long cold storage of meat from chickens fed α-tocopheryl acetate

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supplemented or non supplemented diet (containing natural α-tocopherol) or with synthetic

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antioxidants and carotenoid added can affect fatty acid profile in fat extracted from meat. It

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may also change sensory indices of meat as compared to short cold stored meat. At the dietary

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level 15.9 mg kg-1 of natural α-tocopherol from feed components the similar quality of stored

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air proof packed breast meat was stated as when α-tocopheryl acetate was added to this diet.

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Natural vitamin E content therefore seems to be sufficient.

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References

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Bailey C.A.,. Srinivasan L.J., McGeachin R.B. (1996). The effect of Ethoxyquin on tissue

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peroxidation and immune status of single Comb White Leghorn cockerels. Poultry Sci.,

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75: 1109-1112.

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Bartov I., Bornstein S., 1977. Stability of abdominal fat and meat of broilers: Relative effects of vitamin E, Butylated Hydroxytoluene and Ethoxyquin. Brit. Poultry Sci., 18, 59-68

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Bartov I., Bornstein S. (1981). Stability of abdominal fat and meat of broilers: Combined

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effect of dietary vitamin E and synthetic antioxidants. Poultry Sci.: 60, 1840-1845.

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Baryłko-Pikielna N. (1975). Sensory analysis of meat (in Polish). WNT. Warszawa.

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Coetzee G.J.M. Hoffman L.C. (2001). Effect of dietary vitamin E on the performance of

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broilers and quality of broiler meat during refrigerated and frozen storage. South African

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J. Anim. Sci., 31: 158-173.

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Folch J., Lees M., Stanley G.H.S. (1957). A simple method for the isolation and purification of total lipids from animal tissues. J. Biol. Chem., 226: 497-509. Gray J.I., Gomaa E.A., Buckley D.J. (1996). Oxidative quality and shelf life of meat. Meat. Sci., 43: 111S-123S. Kingsley G.R., Graham P.P., Young W. (1978). Effect of frozen storage and dry-curing on ham trigliceride fatty acids. J. Food Sci., 43, 479-498.

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Koreleski J., Świątkiewicz S. (2006). Effect of stabilized fish oil supplementation and storage

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on changes of fatty acid profile, TBARS content and sensoric properties of breast meat

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of broiler chickens. Polish J. Nat. Sci., Suppl. 3: 421-426.

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Lin C.F., Asghar A., Gray J.I., Buckley D.J., Crackel R.L., Flegal C.J. (1989). Effect of

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oxidized dietary oil andantioxidant suuplementation on broiler growth and meat

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stability. Brit. Poultry Sci., 30: 855-864.

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Manz U.Y., Philipp K. (1981). A method for the routine determination of tocopherols in

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animal feed and human foodstuffs with the aid of high performance liquid

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chromatography. Int. J. Vitamin. Nutr. Res., 51: 342-348.

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Maraschiello C., Sarraga C., Reguiero J.A. (1999). Glutathione peroxidase activity, TBARS,

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and alpha-tocopherol in meat from chickens fed different diets. J. Agric. Food Chem.,

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47: 867-872.

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Miteva E., Bakalivanova T. (2006). Changes in the fatty acid composition of dietetic readyto-cook chicken meat products during frozen storage. Nahrung, 31, 10: 981-986.

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Morrisey P.A., Brandon S., Buckley D.J., Sheehy P.J.A., Frigg M. (1997). Tissue content of

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α-tocopheryl acetate supplement for various periods pre-slaughter. Brit. Poultry. Sci.,

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38: 84-88.

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Morrison W.R., Smith L.A. (1964). Preparation of fatty acid esters and dimethylacetals from lipids with boron fluoride-methanol. J. Lipid Res., 5: 600-608.

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Pikul J., Leszczyński A., Kummerow F.A. (1989). Evaluation of three modified TBA

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methods for measuring lipid oxidation in chicken meat. J. Agr. Food Chem., 37: 1309-

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1317.

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Salih A.M., Smith D.M., Dawson L.E. (1987). Modified extraction 2-thiobarbituric acid method for measuring lipid oxidation in poultry. Poultry Sci., 66, 1483-1488.

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Woodall A., A., Britton G., Jackson M.J. (1996). Dietary supplementation with carotenoids:

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effect on α-tocopherol levels and susceptibility of tissues to oxidative stress. Brit. J.

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Nutr., 76: 307-317.

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Jerzy Koreleski, Sylwester Świątkiewicz

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Wpływ dodatku do paszy witaminy E, przeciwutleniacza i syntetycznego karotenoidu

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na zmiany jakości mięsa piersi kurcząt podczas przechowywania

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STRESZCZENIE

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Doświadczenie przeprowadzono na kurczętach rzeźnych Cobb w okresie od 22 do 42

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dnia życia, które przydzielono do 4 grup doświadczalnych. Stosowano mieszanką paszową

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zawierającą 15,9 mg witaminy E (grupa I), lub z dodatkiem 150 mg octanu dl-α-tocopherylu

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(grupa II), 48,6 mg mieszaniny przeciwutleniaczy BHA+EQ+BHT (grupa III), lub 40 mg/kg

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estru kwasu β-apo-8-karotenowego (grupa IV). W 42 dniu życia, po 4 reprezentatywne dla

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każdej grupy kogutki i kurki dekapitowano, preparowano mięśnie piersiowe, dzielono na

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dwie części, szczelnie pakowano i zamrażano (-20oC). W jednej części mięsa, w okresie do 2

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tygodni analizowano skład kwasów tłuszczowych w lipidach oraz zawartość substancji

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reagujących z kwasem tiobarbiturowym (TBA-RS) i witaminy E. Po ugotowaniu mięsa

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wykonano również jego analizę sensoryczną. Drugą część mięsa trzymano w zamrożeniu

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(-20oC) przez okres 6 miesięcy i także poddano analizie chemicznej i sensorycznej. Skład

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kwasów tłuszczowych lipidów mięsa analizowano chromatograficznie, α-tokoferol oznaczano

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przy użyciu HPLC, a zawartość TBA-RS wyrażono w mg aldehydu malonowego w 1 kg.

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Podczas przechowywania obniżył się w lipidach mięsa udział nasyconych kwasów

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tłuszczowych (SFA), przy równoczesnym wzroście poziomu wielonienasyconych kwasów

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tłuszczowych (PUFA). Zwiększył się stosunek PUFA:SFA oraz nastąpiło obniżenie stosunku

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PUFA n-6:n-3 (7.7 vs 5.5), natomiast zawartość witaminy E i TBA-RS w mięsie nie uległa

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istotnej zmianie. We wszystkich grupach po okresie przechowywania nastąpiło pogorszenie

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zapachu gotowanego mięsa, natomiast pogorszenie smaku stwierdzono u kurcząt

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otrzymujących w paszy syntetyczne przeciwutleniacze i syntetyczny ksantofil. Przy

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zawartości 15.9 mg kg -1 naturalnego α-tokoferolu pochodzącego z komponentów mieszanki

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paszowej uzyskano podobną jakość mięsa przechowywanego w zamrożeniu i szczelnym

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opakowaniu jak po dodatkowym uzupełnieniu tej paszy octanem α-tokoferylu.

Table 1. Composition of basal diet, g · kg-1 Item

Content

Maize

573.5

Soyabean meal

335

Rape seed oil

40

Fish fat (liquid)

10

Limestone

11.5

Dicalcium phosphate

18

NaCl

3

DL-methionine (99%)

2

L-lysine HCl (78 %)

2

Vitamin-mineral premix

1

5

Crude protein analyzed

201

ME. (MJ · kg-1)

12.82

Lys 3

12.0

Met 3

5.10

Ca

3

P total

9.30 3

7.15

P available 3

4.10

α-tocopherol (mg · kg ) analyzed -1 4

1

15.95

contained no antioxidants and vitamin E, supplied to 1 kg of diet: IU: vit. A 12 000; D 3 3250; mg: K3 2.25; B1 2; B2 7.25; B6 4.25; B12 0.03; biotin 0.1; Ca-pantotenate 12; niacine 40; folic acid 1.0; choline-Cl 450; Mn 100; Zn 65; Fe 65; Cu 15; J 0.8; Se 0.25 and Co 0.4 2 according to European table of energy values for poultry feedstuffs (1989) as a sum of ME content of feed components calculated on base of nutrients content 3 calculated from tables of feed composition 4 after supplementing with 150 mg α-tocoferyl acetate the diet was analyzed to contain 160.69 mg · kg -1 vitamin E in total

Table 2. Effect of storage on selected fatty acids profile in lipid fraction of breast meat (% of total fatty acids) C16:0

C18:0

C18:1

C18:2 (LA)

C18:3 (ALA)

C20:4

C20:5 (EPA)

C22:6 (DHA)

I, Control: short stored long stored SEM

17.1b 11.8a 0.97

6.01a 7.86b 0.32

27.6a 29.8b 0.51

35.8b 31.9a 0.75

2.52b 2.26a 0.06

3.78a 7.76b 0.70

0.501a 0.723b 0.043

2.35a 4.28b 0.45

II, Vitamin E: short stored long stored SEM

16.9b 11.7a 0.93

6.82a 9.20b 0.62

25.8 26.2 0.83

36.5b 31.1a 1.05

2.51 2.08 0.15

4.25a 9.56b 1.14

0.501a 0.792b 0.064

2.62a 5.98b 0.67

III, BHA+EQ +BHT: short stored long stored SEM

18.7b 13.4a 1.01

6.42a 8.67b 0.42

25.6 27.4 0.60

36.1b 31.9a 1.09

2.17 1.90 0.13

4.09a 8.56b 0.88

0.486a 0.689b 0.053

2.33a 4.60b 0.63

IV, Synth. yellow xanthophyll: short stored long stored SEM

17.5b 12.1a 0.98

6.27a 8.55b 0.40

27.6a 29.4b 0.39

36.1b 32.4a 0.87

2.49 2.24 0.08

3.17a 7.34b 0.70

0.476a 0.737b 0.060

1.84a 3.85b 0.39

Treatment

a, b

- within groups values in the same columns with different letters differ significantly at P≤0.05

Table 3.

Effect of storage on profile of fatty acid categories in lipid fraction of breast meat (% of total fatty acids) SFA

UFA

PUFA

PUFA n-6

PUFA n-3

I, Control: short stored long stored SEM

24.4b 20.1a 0.83

75.6a 79.9b 1.09

46.1 48.8 0.75

39.7 39.7 0.44

5.37a 7.26b 0.43

3.10a 4.00b 0.18

1.89a 2.45b 0.12

7.42b 5.59a 0.37

II, Vitamin E: short stored long stored SEM

25.1b 21.4a 0.89

74.9a 78.6b 1.21

47.5a 51.3b 0.87

40.9 40.7 0.45

5.63a 8.83b 0.60

3.01a 3.70b .16

1.91a 2.41b 0.10

7.26b 4.66a 0.45

III, BHA+EQ +BHT: short stored long stored SEM

26.7b 22.5a 0.93

73.3a 77.5b 1.11

46.1a 49.1b 0.72

40.3 40.5 0.51

4.89a 7.18b 0.60

2.76a 3.46b 0.15

1.74a 2.19b 0.10

8.26b 6.01a 0.53

IV, Synth. yellow xanthophyll: short stored long stored SEM

25.4b 21.1a 0.84

74.6a 78.9b 1.34

45.1a 48.4b 0.74

39.4 39.8 0.61

4.81a 6.83b 0.39

2.94a 3.76b 0.16

1.78a 2.30b 0.11

8.21b 5.93a 0.46

Treatment

a, b

UFA/SFA PUFA/SFA ratio ratio

- within groups values in the same columns with different letters differ significantly at P≤0.05

n-6/n-3 ratio

Table 4.

Effect of storage on vitamin E and TBARS content in breast meat and sensoric properties of boiled meat

Treatment

Vitamin E, mcg · g-1

TBARS, mg of malondialdehyde · kg-1

Flavour of meat, points

Taste of meat, points

Tenderness of meat, points

Juiceness of meat, points

I, Control: short stored long stored SEM

2.11 1.72 0.13

0.373 0.381 0.017

4.65b 4.37a 0.052

4.57 4.42 0.052

4.65 4.47 0.078

4.55 4.42 0.069

II, Vitamin E: short stored long stored SEM

7.05 6.45 0.26

0.347 0.389 0.025

4.72b 4.37a 0.063

4.62 4.45 0.060

4.70 4.53 0.052

4.65 4.42 0.060

III, BHA+EQ+BHT: short stored long stored SEM

2.03 1.76 0.12

0.436 0.447 0.015

4.62b 4.24a 0.067

4.55 4.25 0.067

4.60 4.39 0.074

4.50 4.39 0.082

IV, Synth. yellow xanthophyll: short stored long stored SEM

2.18 1.83 0.15

0.430 0.461 0.015

4.70b 4.06a 0.080

4.47b 4.00a 0.071

4.65b 4.31a 0.070

4.55b 4.23a 0.074

a, b

- within groups values in the same columns with different letters differ significantly at P≤0.05