Effect of dietary mixture of gallic acid and linoleic acid on antioxidative potential and quality of breast meat from broilers

Meat Science 86 (2010) 520–526

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Meat Science j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / m e a t s c i

Effect of dietary mixture of gallic acid and linoleic acid on antioxidative potential and quality of breast meat from broilers Samooel Jung a, Jun Ho Choe a, Binna Kim a, Hyejeong Yun a, Zbigniew A. Kruk b, Cheorun Jo a,⁎ a b

Department of Animal Science and Biotechnology, Chungnam National University, Daejeon, 305-764, Republic of Korea Department of Animal Science, School of Agriculture Food and Wine, The University of Adelaide, South Australia 5371, Australia

a r t i c l e

i n f o

Article history: Received 16 October 2009 Received in revised form 31 May 2010 Accepted 8 June 2010 Keywords: Broiler Gallic acid Linoleic acid Antioxidative potential Breast meat quality

a b s t r a c t The effect of dietary mixture of gallic acid and linoleic acid (MGL) on the antioxidative potential and quality of breast meat from broilers was investigated. Broilers during the 22–36 days on trial received 3 dietary treatments: 1) control (commercial finisher diet), 2) 0.5% MGL (gallic acid:linoleic acid = 1 M:1 M), and 3) 1.0% MGL. The feed efficiency, DPPH radical scavenging activity, ABTS+ reducing activity, reducing power, TBARS, and total phenolic content in the breast from the broilers improved significantly by 1.0% MGL dietary treatment. Arachidonic and docosahexaenoic acids were higher in the broilers fed both levels of MGL diets. In addition, water holding capacity of the breast was enhanced by the 1.0% dietary MGL treatment and was accompanied by a slight antimicrobial activity (1 decimal reduction) during storage. In conclusion, 1.0% dietary supplementation with MGL can improve the antioxidative potential, and nutritional and functional qualities of broiler breast meat. © 2010 The American Meat Science Association. Published by Elsevier Ltd. All rights reserved.

1. Introduction There has been an increased interest in food containing high amount of polyunsaturated fatty acids (PUFAs) because PUFAs are considered as functional ingredients to prevent coronary heart disease and other chronic diseases (Krauss et al., 2001; Russo, 2009). Linoleic acid (LA; C18:2n-6) is one of the essential fatty acids and the primary precursor of all n-6 PUFAs (Russo, 2009). It is converted to arachidonic acid (AA; C20:4n-6) in animal tissues (Smith, 2008) and has been shown to possess anti-inflammatory effect by decreasing the secretion of interleukin (IL)-6 and -1β and the tumor necrosis factor α (Zhao et al., 2005). Previous study reported that high levels of dietary LA suppressed lymphocyte proliferation in rats (Yaqoob, Newsholme, & Calder, 1994). For this reason, several studies had been conducted to increase the content in PUFAs in chickens and eggs by using dietary fat sources such as natural oil containing PUFAs and LA (Crespo & Esteve-Garcia, 2002; Kim et al., 2007). However, PUFAs are prone to oxidation since they are the first targets for free radical strike at initiating peroxidation (Scislowski, Bauchart, Gruffat, Laplaud, & Durand, 2005). The oxidation products lead to deterioration of food quality such as flavor, color, texture and nutritional value, and may be responsible for tissue and organ damage (Mielnik, Olsen, Vogt, Adeline, & Skrede, 2006; Priscilla & Prince, 2009). The notable strategies for diminishing lipid oxidation of meat utilise diets containing antioxidants. Currently, the interest in natural

⁎ Corresponding author. Tel.: +82 42 821 5774; fax: +82 42 825 9754. E-mail address: [email protected] (C. Jo).

antioxidants has increased because they are considered to be safer than the synthetic antioxidants, and have greater application potential for consumers' acceptability, palatability, stability and shelf-life of meat products (Kang et al., 2008; Naveena, Sen, Vaithiyanathan, Babji, & Kondaiah, 2008; Park & Kim, 2008). However, the use of natural antioxidants in animal nutrition could be restricted due to the low bioavailability of polyphenols. Moreover, many types of polyphenols can lose a part of their antioxidant capability in vivo (Manach, Scalbert, Morand, Rémésy, & Jiménez, 2004). Previous studies have shown that the negative outcome of lipid oxidation in chicken meat and eggs was diminished by the use of diets containing antioxidants such as medicinal herb mix and grape pomace (Goñi et al., 2007; Jang, Liu et al., 2008a; Liu et al., 2009) which are natural antioxidants rich in polyphenols. The pork meat derived from the pig fed oleoresins of rosemary did not show any antioxidant effect (Lopez-Bote, Gray, Gomaa, & Flegal, 1998). In the lights of these inconsistent results, many researchers have been investigating a variety of other natural antioxidants that possesses a superior protective activity. Gallic acid (GA) is a representative of natural polyphenolics found in wine, grapes as well as tea (Hogan, Zhang, Li, Zoecklein, & Zhou, 2009; Kim et al., 2006). GA, a metabolite of propyl gallate, is known to possess several pharmacological and biological activities such as strong antioxidant, anti-carcinogenic, anti-mutagenic, anti-allergic, and antiinflammatory (Jo, Jeong, Lee, Kim, & Byun, 2006; Schlesier, Harwat, Bohm, & Bitsch, 2002). Jo et al. (2006) synthesized a novel compound from GA and LA (octadeca-9,12-dienyl-3,4,5-hydroxybenzoate, GA–LA) and reported that synthesized GA–LA possessed greater biological function than the GA itself, including antioxidative effect, tyrosinase

0309-1740/$ – see front matter © 2010 The American Meat Science Association. Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.meatsci.2010.06.007

S. Jung et al. / Meat Science 86 (2010) 520–526

inhibition, and anti-inflammatory effects. Jang et al. (2008b, 2009) reported that GA–LA had a strong and synergistic inhibition effect on cancer cell proliferation (in vitro) and suggested that synthesized GA–LA may be a useful functional material for the food, pharmaceutical, and cosmetic industries. Furthermore, not only the synthesized GA–LA but also the mixture of GA and LA (MGL) successfully protected against cardiovascular diseases that result from hyperlipidemia. The MGL mixture showed no adverse effects when using high-fat diet to induce obesity in mice (in vivo) (Jang, Srinivasan, et al., 2008b). Since the possibility of using the synthesized GA–LA on the industrial scale is not economically viable, dietary MGL can serve as a candidate that has a potential to improve broilers performance and consequently broiler meat quality including its antioxidative potential. The objective of this study was to evaluate the effect of dietary MGL mixture supplementated to broilers on their performance, antioxidative potential, and quality of the broiler's breast fillet. 2. Materials and methods 2.1. Animal and experimental design A total of 90 one-day old male and female broilers (Ross strain) were obtained from a commercial hatchery. Broilers were randomly assigned to 9 pens under the standard condition of temperature, humidity, and ventilation, and 24 h fluorescent lighting for the entire experimental period. Broilers had at libitum access to water and diet, and were fed a commercial broiler starter (0–6 days) and grower (7– 21 days) diets. At the end of the week 3, broilers were weighed and reassigned to 3 different dietary treatments based on the average weight. Each treatment had 3 replicates containing 10 broilers in each replicate. During the 22–36 days of the experimental period broilers were fed the following diets: 1) control [commercial finisher diet], 2) 0.5% MGL (w/w, GA:LA = 1 M:1 M), and 3) 1.0% MGL. The control diet contained approximately 20% of crude protein, ∼4% of crude fiber, 3100 ME kcal/kg and was a typical commercial diet produced for broilers (Chunhajeil Feed Co., Daejeon, Korea). The treatment diets were based on the commercial diet supplemented with 0.5% or 1% of MGL. GA and LA were purchased from Yakuri Chemical Co. (Osaka, Japan) and Sigma-Aldrich Co. (St. Louis, MO, USA), respectively. At the end of the experimental period, broilers were weighed, and feed efficiency was calculated. At day 36, 7 broilers from each pen were slaughtered in the research unit by the carotid amputation. The carcasses were removed of feathers, eviscerated, vacuum packed and were stored in a deep freezer at −50 °C until required for analysis. Broilers care facilities and the procedures performed met or exceeded the standards established by the Committee for Accreditation of Laboratory Animal Care at Chungnam National University, Korea. The study was conducted in accordance with recommendations described in “The Guide for the Care and Use of Laboratory Animals” published by the institutional Animal Care and Use Committee (IACUC) of Chungnam National University. 2.2. Blood collection and analysis At the end of the experiment, 1 broiler from each pen was randomly selected. Blood was collected via wing veinpuncture using syringe (Greenject-5, Doowon MediTec, Co., Gimje, Korea), placed into vacuum tubes containing ethylenediaminetetraacetic acid and stored at −4 °C. Samples were centrifuged (Union 32R, Hanil Co., Ltd., Inchun, Korea) at 1500 ×g for 15 min and then serum was separated. The separated serum was used for analysis of the content of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), total cholesterol (T. chol), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C).

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2.3. Broiler meat quality The frozen broiler carcasses were thawed in a refrigerated condition (4 °C) for 48 h and the breast fillets were dissected. 2.3.1. pH pH was measured in the breast meat of broiler using a pH meter (SevenGo, Mettler-Toledo Inti, Inc. Schwerzenbach, Switzerland) equipped with an insertion glass electrode after calibration using buffers at pH 4.01, 7.00, and 9.21 at room temperature. 2.3.2. Water holding capacity One gram of the minced breast meat of broiler was placed on a round filter paper (No. 4, Whatman Ltd. Kent, UK). The filter paper with meat was placed into the centrifuge tube and was centrifuged (CR 20B2, Hitachi Koki Co., Ltd. Fukuoka, Japan) at 6710 × g for 10 min. The released water absorbed into the filter paper was weighed and calculated as a percentage of the initial moisture of meat. 2.4. Measurement of antioxidative potential Breast fillets (3 g) were homogenized (T25b, Ika Works (Asia)., Sdn, Bhd, Malaysia) in 15 ml of distilled water at 1130 ×g for 1 min. Chloroform (10 mL) was added to the homogenates and the mixture was shaken vigorously 2–3 times. Lipids and the aqueous supernatant were separated by centrifugation (Hanil) at 2090 ×g for 15 min. The supernatant was used for the measurement of total phenolic content, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, 2,2-azinobis-(3 ethylbenzothiazoline-6-sulfonic acid) (ABTS+) reducing activity, and reducing power. 2.4.1. Total phenolic content Total phenolic content was estimated by the Folin–Ciocalteu method (Subramanian, Padmanaban, & Sarma, 1965). A 0.1 mL aliquot was added to the Folin–Ciocalteu reagent (0.2 mL), followed by the addition of 3 mL sodium carbonate solution (5%). The reaction mixture was vortexed and the absorbance was measured with a spectrophotometer (DU 530, Beckman Instruments Inc., Fullerton, CA, USA) at 765 nm after incubation for 1 h at 23 °C. The quantification of phenolics was based on the standard curve generated with the use of gallic acid, and expressed as gallic acid equivalent. 2.4.2. DPPH radical scavenging activity DPPH radical scavenging activity was estimated according to the method of Blois (1958) with slight modifications. A 200 μL aliquot was added to 800 μL distilled water and 1 mL methanolic DPPH solution (0.2 mM). The mixture was vortexed and left to stand at room temperature (20–22 °C) for 30 min. A tube containing 1 mL of distilled water and 1 mL of methanolic DPPH solution (0.2 mM) served as the control. The absorbance of the solution was measured at 517 nm using a spectrophotometer (Beckman). The percentage of DPPH radical scavenging was obtained from the following equation: Radical scavenging activity = ½1−ðabsorbance of sample = absorbance of controlÞ × 100:

2.4.3. ABTS+ reducing activity ABTS+ reducing activity was determined as the method described by Erel (2004). ABTS was dissolved in distilled water to a 7 mM concentration. The ABTS radical cation was produced by acting the ABTS stock solution with 2.45 mM potassium persulfate (final concentration) in the dark at room temperature for 12–16 h to allow the completion of radical generation. This solution was then diluted with ethanol so that its absorbance was adjusted to 0.70 ± 0.02 at 734 nm.

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The diluted ABTS+ solution (3 mL) were added to 20 μL aqueous supernatant and the absorbance was measured by a spectrophotometer (Beckman) at 734 nm using ethanol as a blank. The percentage inhibition was calculated by the following equation: þ

ABTS reducing activityð%Þ = ½ðabsorbance of control–absorbance of sampleÞ  absorbance of control × 100:

2.4.4. Reducing power The reducing power was determined according to the method of Oyaizu (1986). A 200 μL aliquot was mixed with 500 μL sodium phosphate buffer (0.2 M, pH 6.6) and 500 μL potassium ferricyanide (1%), and the resultant mixture was incubated at 50 °C for 20 min. After addition of 2.5 mL trichloroacetic acid [TCA, (10%)], the mixture was centrifuged (Hanil) at 2200 ×g for 10 min. The upper layer (500 μL) was mixed with 500 μL distilled water and 100 μL ferric chloride (0.1%), and the absorbance was measured at 700 nm using a spectrophotometer (Beckman): higher absorbance indicates higher reducing power. 2.4.5. 2-Thiobarbituric acid-reactive substances (TBARS) Each meat sample (5 g) in 15 mL distilled water was homogenized (Ika Works) at 1130 ×g for 1 min. Sample homogenate (1 mL) was transferred to a test tube and lipid oxidation was determined as the TBARS value by using the method described by Ahn, Olson, Jo, Love, and Jin (1999). Briefly, 50 μL butylated hydroxyanisol (7.2%) and 2 mL TBA–TCA solution (20 mM TBA in 15% TCA) were added to the test tube. Tubes were heated (90 °C) in a boiling water bath for 30 min, cooled, and then centrifuged at 2090 ×g for 15 min. Absorbance of the supernatant was measured at 532 nm with a spectrophotometer (Beckman). TBARS value was reported as mg malondialdehyde per kg meat.

37 °C for 24 h. Colony forming units (CFU) per gram were counted, at a dilution giving 30–300 CFU per plate. 2.7. Volatile basic nitrogen (VBN) Measurement of VBN in the breast meat of broiler was done according to Conway (1950). Each meat sample (3 g) was homogenized for 1 min with 3 mL distilled water and 6 mL TCA (10%), and then centrifuged (Hanil) at 2090 ×g for 15 min. The supernatant was filtered using a filter paper (No. 4, Whatman), and the filtrate was placed in a test tube and made up to a final volume of 30 mL with 5% TCA. A volume of 0.01 N boric acid as a VBN absorber was placed in the inner section of a Conway micro-diffusion cell (Sibata Ltd. Tokyo, Japan). A 1 mL sample solution and 1 mL saturated K2CO3 was also placed into the outer section of the same cell and the lid was immediately closed. A 5% TCA solution was used as blank. The cell was incubated at 37 °C for 120 min, and then titrated against 0.02 N sulfuric acid. The concentration of VBN was calculated as ammonia equivalent using the following equation: VBN valueðmg%Þ = ½0:28 × ðtitration volume of sample solution–titration volume of blankÞ × 10 × 100:

2.8. Statistical analysis A total of 9 pens with 10 broilers per pen were used for this experiment with 3 replications for each treatment. Analysis of variance was performed using the raw data, and the mean values and standard errors of the means (SEM) were calculated by the Statistical Analysis System (SAS, 2000). Differences among the means were determined by the Duncan's multiple range test with a significance defined at P b 0.05.

2.5. Fatty acid composition

3. Results and discussion

Total lipid of samples was extracted by using chloroform–methanol (2:1, v/v) according to the procedure of Folch, Lees, and Sloane Stanley (1957). The fatty acid methyl esters were prepared from the extracted lipids with BF3-methanol (Sigma-Aldrich). The fatty acid methyl esters were, then, separated on a gas chromatograph (Agillent GC 6890N, Palo Alto, CA, USA) equipped with a mass selective detector (MSD). A split inlet (split ratio, 50:1) was used to inject samples into a HP-5MS capillary column (30 m × 0.25 mm × 0.25 μm), and ramped oven temperature was used (150 °C for 3 min, increased to 180 °C at 2.5 °C/min and maintained for 5 min, then increased to 220 °C at 2.5 °C/min and maintained for 25 min). Inlet temperature was 210 °C. Helium was the carrier gas at constant flow of 0.7 mL/min. The temperature of the mass spectrometer (MS) source, MS quadrupole, and the transfer line into the MS were 230, 150, and 280 °C respectively. The fatty acid composition was identified by a mass spectrum database (NIST Library, mass spectral search program, version 5.0, Ringoes, NJ, USA).

3.1. Broiler performance

2.6. Microbiological analysis Media for the enumeration of the total aerobic bacteria were prepared by tryptic soy agar (Difco Laboratories, Detroit, MI, US; PCA) to provide the optimum conditions for the growth of halophiles. The breast meat (10 g) with 100 mL saline solution (0.85%, NaCl) was homogenized for 2 min using a stomacher homogenizer (BagMixer 400, Interscience, Paris, France) and the homogenate was serially diluted 10-fold with the saline solution. Each diluent (100 μL) was spread in triplicate on each agar plate and the plate was incubated at

The effect of dietary MGL on feed intake, body weight gain, and feed conversion is shown in Table 1. Feeding 1.0% MGL significantly improved the daily weight gain and decreased the feed to gain ratio, whereas no effect on daily feed intake was observed. Feed intake, growth rate, and feed efficiency of the poultry are impaired by environmental stress, mainly heat stress (Bartlett & Smith, 2003). Improved feed efficiency of quail was reported by supplementation of vitamin C as an antioxidant which affected the utilization of dietary nutrients (Sahin et al., 2006). Previous studies reported that dietary antioxidants, such as vitamin C, E, flavonoids, and phenolics, can reduce oxidative damage in animals which is generated by different stress sources (Bagchi et al., 1999; Brisibe et al.,

Table 1 Effect of the dietary supplementation of the mixture of gallic acid and linoleic acid on feed intake, daily weight gain, and feed conversion of broiler during finisher (22–36 days) period. Item

Feed intake (g/bird per d) Weight gain (g/bird per d) Feed/gain (g/g) 1

Diet Control

0.5% MGL

1.0% MGL

SEM1

164.9 79.6b 2.06a

164.5 79.8b 2.06a

158.0 80.8a 1.95b

2.30 0.17 0.024

Standard errors of mean (n = 9). Different letters within the same row differ significantly (P b 0.05).

a,b

S. Jung et al. / Meat Science 86 (2010) 520–526

2009). Therefore, the dietary supplementation of MGL, which contained gallic acid, natural antioxidant, may improve feed efficiency by reducing oxidative damage in broilers.

Table 3 pH, water holding capacity (%), and total phenolic contents (mg gallic acid equivalent/g meat) of the breast meat from the broiler fed the mixture of gallic acid and linoleic acid after 36 days. Treatment

3.2. Serum lipid profile Serum GOT (also known as aspartate aminotransferase, AST) and GPT (known as alanine aminotransferase, ALT) were used as the biochemical indicators for liver function. The increase of these enzyme activities indicates the liver damage (Han, Huang, Li, Jiang, & Xu, 2008). Khan, Hussain, and Khan (2006) reported that the change of serum GOT and GPT were reflected by the adverse effect of chicken fed formalin. In the present study, serum GOT and GPT were not affected by dietary MGL (Table 2). Dietary MGL did not significantly influence the concentration of serum total cholesterol, HDL-C, and LDL-C of broilers. However, supplementation of 1.0% MGL significantly reduced TG by 55.54% when compared with control. These results are in agreement with the previous study that showed the reduction of serum TG level of mice that were fed a high-fat diet supplemented with MGL (Jang, Srinivasan, et al., 2008b). Tai et al. (2005) reported that the high intake of dietary polyunsaturated fatty acids can reduce serum TG level by modulating genes involved in lipid metabolism (peroxisome proliferator activated receptor α). High serum TG concentration was recognized as an independent risk factor for atherosclerosis and closely correlated with the increase of the risk of coronary heart disease (Smith et al., 2004). Therefore, our result suggests that dietary MGL is an effective health promoting agent to improve lipid metabolism in live animals.

3.3. pH and water holding capacity (WHC) The final pH of the muscle is very important due to the positive correlation with WHC during the conversion of muscle to meat (Bee, Anderson, Lonergan, & Huff-Lonergan, 2007). The decrease of meat pH to near isoelectric point results in the decrease of WHC (Swatland, 2008). Dietary MGL had no significant effect on pH of the breast meat from broilers postmortem (Table 3). However, WHC of the breast meat from broilers was significantly increased by the 1.0% dietary MGL treatment when compared with control. Huff-Lonergan, and Lonergan (2005) reported that WHC is not only influenced by the pH but also postmortem proteolysis which begun with the activation of μcalpain, that may be inactivated by oxidation. Previous studies have indicated that antioxidants in meat can affect the proteolysis (Rowe, Maddock, Trenkle, Lonergan, & Huff-Lonergan, 2004). Therefore, it can be thought that the dietary MGL improved antioxidative system in chicken breast meat, resulting in the improvement of WHC.

Control 0.5% MGL 1.0% MGL SEM1

GOT (AST) U/L

GPT (ALT) U/L

T. chol mg/dL

TG mg/dL

HDL-C mg/dL

LDL-C mg/dL

239 214 254 31.6

3 4 5 0.7

139 151 152 13.3

81x 51xy 36y 10.4

114 120 131 9.1

20 26 26 3.8

Water holding capacity

Total phenolics

y

5.94 5.94 5.97 0.013

1.48y 1.50xy 1.54x 0.011

51.66 54.74y 60.40x 1.194

1

Standard errors of mean (n = 9). Different letters within the same column differ significantly (P b 0.05).

x,y

3.4. Antioxidative activity and total phenolic content Various analyses were conducted to evaluate the antioxidative effect of dietary MGL (Table 4). DPPH is a widely used method for estimating the antioxidative activity. A solution of DPPH, stable free radical, is mixed with an antioxidant that can donate a hydrogen atom to form a stable DPPH-H molecule. Then this reduced form is visualised by the loss of violet color (Molyneux, 2004). The DPPH radical scavenging activity of the breast meat from the broilers fed 1.0% MGL was significantly higher than that of the control during the entire storage period, whereas no significant difference was found in the breast meat of the broilers fed either control or 0.5% MGL diets. The ABTS, which is a stable free radical cation applicable to both lipophilic and hydrophilic antioxidants, has been used to measure total antioxidative activity (Kim & Lee, 2009). The breast meat of the broiler fed 1.0% MGL had significantly higher ABTS+ reducing activity than that of control during the whole storage period except for day 0. The ABTS+ reducing activity of the breast meat from the broilers fed 0.5% MGL was significantly higher than that of the control at storage in days 2 and 7. However, there was no difference at days 0 and 4. The reducing power is another significant indicator of antioxidant activity potential and is based on chelating effect on ferrous ion (Fe3+ to Fe2+) (Hsu, Coupar, & Ng, 2006). The reducing power of the breast meat from the broilers fed 0.5 and 1.0% MGL was significantly greater than that of control at days 4 and 7, but no significant difference was

Table 4 Antioxidative potential of breast meat from broiler fed mixture of gallic acid and linoleic acid stored at 4 °C. Treatment

Storage (day) 4

7

SEM1

DPPH radical scavenging activity (%) 68.18ay Control 67.77ay 69.00axy 0.5 % MGL 70.01axy 70.19bx 1.0 % MGL 72.21ax 0.738 0.362 SEM2

66.60aby 68.28ay 70.48bx 0.564

64.49by 64.48by 67.99cx 0.827

0.716 0.718 0.480

ABTS+ reducing Control 0.5 % MGL 1.0 % MGL SEM2

activity (%) 30.26ab 33.28b 33.28b 1.147

28.22by 31.63bxy 33.05bx 1.277

28.70by 34.05abx 34.43bx 1.393

1.330 1.402 1.155

Reducing power Control 0.5% MGL 1.0% MGL SEM2

0.593a 0.605 0.606b 0.0087

0.597a 0.598 0.656a 0.0191

0.560aby 0.609x 0.627abx 0.0093

0.515by 0.577x 0.563cx 0.0094

0.0161 0.0108 0.0093

TBARS value Control 0.5% MGL 1.0% MGL SEM2

0.29cx 0.23by 0.23cy 0.017

0.32cx 0.27by 0.26by 0.012

0.39bx 0.43ax 0.33ay 0.014

0.49ax 0.47ax 0.35ay 0.014

0.019 0.013 0.009

0

1

Standard errors of mean (n = 9). Glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), total cholesterol (T. chol), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C). x,y Different letters within the same column differ significantly (P b 0.05).

pH

Control 0.5% MGL 1.0% MGL SEMa

Table 2 Effect of dietary supplementation of the mixture of gallic acid and linoleic acid on blood profiles in broiler after 36 days. 2

523

2

1

2

33.53ay 38.47ax 38.52ax 1.369

Standard errors of mean (n = 12). 2(n = 9). Different letters within the same row differ significantly (P b 0.05). x,y Different letters within the same column differ significantly (P b 0.05). a–c

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found at days 0 and 2 of the storage period. These results suggest that breast meat samples from the broilers fed MGL were capable electron donors for neutralizing free radicals. The development of lipid oxidation in the breast meat from broilers was delayed by the 1.0% dietary MGL during the entire storage period. The breast meat from the broilers fed MGL produced low TBARS in spite of the higher content of linoleic acid (LA) in MGL diet. The LA belongs to the PUFA family that has potential to generate several types of free radicals and can accelerate lipid peroxidation (Nagendra Prasad et al., 2009). GA in MGL diet may also play a role in antioxidant activity. These results agree with the other studies which showed that lipid oxidation of the meat from the chickens fed various PUFA levels with α-tocopheryl acetate was inhibited due to the antioxidant activity of α-tocopheryl acetate (Cortinas et al., 2005). Total phenolic content of the breast meat from the broilers fed MGL was shown in Table 3. Breast meat of the broilers fed 1.0% MGL had significantly higher total phenolic content when compared with control, but 0.5% dietary MGL was not different. These results indicate that dietary MGL improved the antioxidative activity of the breast meat. Previous study reported that GA is a polyphenyl natural product (Kim et al., 2006) and may directly combine with free radicals and lead to their inactivation, which in turn may decrease the intracellular concentration of free radicals (Priscilla & Prince, 2009). Schwarz et al. (2009) reported that reactive oxygen species, including free radicals play a key role in the oxidation process that can damage cells, whereas polyphenols have been shown to scavenge free radicals such as superoxide, peroxyl, and hydroxyl radicals, and hence influence on the redox mechanisms that may lead to degenerative diseases conditions such as Alzheimer's, atherosclerosis, diabetes, and certain cancers (Hogan et al., 2009). Nagendra Prasad et al. (2009) suggested that polyphenol content showed highest relations with total antioxidant capacity (R2 = 0.9773), and dietary phenolic source such as oregano, rosemary, sage essential oil, and grape pomace had significant antioxidative activities in lamb and broiler meat (Goñi et al., 2007; Simitzis et al., 2008). 3.5. Fatty acid composition The influence of dietary MGL on fatty acid composition in the breast meat of broilers is shown in Table 5. The concentrations of palmitic acid (C16:0) and oleic acid (C18:1) in the breast meat were significantly lower in broilers fed 1.0% MGL than that of the control. These results are probably due to the endogenous synthesis of these fatty acids in the broiler tissues. The saturated fatty acids (SFAs) in poultry tissues rely upon their presence in the diet and their synthesis in the liver. The SFAs synthesis is inhibited in the liver more during

Table 5 Fatty acid composition in the breast meat from the broiler fed the mixture of gallic acid and linoleic acid. Fatty acids

Treatment Control

C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 C20:4 C22:6 Saturated Monounsaturated Polyunsaturated Unsaturated : saturated n-6 : n-3 1

23.12a 2.56 15.04 33.42a 16.46 0.51b 6.88b 2.00b 38.16a 35.98 25.86b 1.62b 9.27a

0.5% MGL 22.93ab 2.11 14.88 32.66ab 16.16 0.49b 8.06a 2.70a 37.81ab 34.77 27.42a 1.64ab 7.58b

1.0% MGL 22.31b 2.30 14.73 32.36b 16.03 0.75a 8.52a 2.99a 37.04b 34.66 28.29a 1.70a 6.56c

Standard errors of mean (n = 9). a–c Different letters within the same row differ significantly (P b 0.05).

digestion of unsaturated fats than saturated fats (Sim & Qi, 1995). Also, the increase of PUFAs decreased the synthesis of monounsaturated fatty acids (MUFAs) by inhibiting the activity of 9-desaturase complex which is the key enzyme needed to convert SFAs to MUFAs (Pinchasov & Nir, 1992). Arachidonic acid (AA; C20:4) composition in the breast meat was significantly increased by 1.0% MGL. It might be due to the presence of LA in MGL diet which is a precursor of AA (Smith, 2008). Previous studies have shown that the fatty acid profiles of broilers meat reflect their intake from the diet. When feeding pigs and broilers with sunflower diet containing LA, the diet was able to increase LA and AA levels in meat (Crespo & Esteve-Garcia, 2002; Guillevic, Kouba, & Mourot, 2009). Docosahexaenoic acid (DHA, C22:6, n-3) levels in the breast meat from the broilers fed 0.5 and 1.0% MGL were also higher than that of the control. This can be explained by the antioxidative effect of GA in dietary MGL treatment. DHA is responsible for hypolipidemic and neuroprotective effects (Rodrigues de Turco et al., 2002). However, DHA is very sensitive to oxidation that can change their pharmacological properties (Judé et al., 2003). Therefore, the dietary MGL may inhibit the oxidation of DHA. The PUFAs/SFAs ratio in the breast meat from broilers was increased by 1.0% dietary MGL supplementation. These results can be attractive to the consumers as high PUFAs/SFAs ratio has a positive health benefit for humans, mainly in protection against cardiovascular disease (Krauss et al., 2001). 3.6. Antimicrobial effect and volatile basic nitrogen (VBN) The growth of total aerobic bacteria in the breast meat from broilers was not influenced by dietary MGL until storage day 4 (Table 6). However, the final population of total aerobic bacteria at storage day 7 in the breast meat from the broiler fed 1.0% MGL was 4.08 log CFU/g, while that of control was 5.52 log CFU/g. This result suggested that 1.0% dietary MGL may have antimicrobial effect in the breast meat. Dietary MGL was consisted of LA and GA which are well known to possess antimicrobial activity (Rodriguez-Vaquero, Alberto, & Manca de Nadra, 2007). Chanwitheesuk, Teerawutgulrag, Kilburn and Rakariyatham (2007) reported that GA exhibited the activity against Salmonella typhimurium and Staphylococcus aureus. However, antimicrobial activity of GA is not apparent in vivo. The amount of VBN content was increased during storage period (Table 6). Dietary MGL did not significantly affect the VBN content in the breast meat until day 4 of storage. However, the VBN content in the breast meat from the broilers fed 1.0% MGL was lower than that of control at storage in day 7 (16.80 vs 19.60 mg%). This result represents a similar trend to the growth of total aerobic bacteria in the breast meat. The VBN content has been considered as spoilage indicator, and Table 6 Number of total aerobic bacteria (log CFU/g) and volatile basic nitrogen (mg%) in the breast meat from broiler fed mixture of gallic acid and linoleic acid. Treatment

SEM1 0.150 0.175 0.096 0.261 0.123 0.029 0.138 0.081 0.236 0.368 0.269 0.014 0.187

Storage (day) 0

2

Total aerobic bacterial count (log CFU/g) 3.75bx Control 3.26b 3.31by 0.5% MGL 3.35b 1.0% MGL 3.60c 3.75bx 0.123 0.092 SEM2 Volatile basic nitrogen (mg%) Control 14.00c 0.5% MGL 14.00c 1.0% MGL 13.06b 0.538 SEM2 1

13.06c 11.20d 13.06b 0.762

4

SEM1

7 3.90b 3.68b 3.75b 0.085

16.80b 16.80b 15.86a 0.538

5.52ax 4.99axy 4.08ay 0.211

19.60ax 18.66ax 16.80ay 0.538

Standard errors of mean (n = 12). 2(n = 9). Different letters within the same row differ significantly (P b 0.05). x,y Different letters within the same column differ significantly (P b 0.05). a–d

0.159 0.174 0.027

0.466 0.466 0.808

S. Jung et al. / Meat Science 86 (2010) 520–526

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