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effect. Green tea has over 200 bioactive compounds and contains over 300 different substances (Labdar, 2010). Numerous in vitro and in vivo studies on green tea preparations have demonstrated the antimutagenic, anticarcinogenic and antioxidant properties of the polyphenolic compounds, which are mainly composed of four types of catechin: epicatechin, epigallocatechin, epigallocatechin gallate and gallocatechin (Yamamoto et al., 1997). The predominant green tea component has been shown to improve body weight gain and feed efficiency in pig (Ko et al., 2008), cattle (Sarker et al., 2010) and broiler (Biswas and Wakita, 2001). In addition, this component maintains microflora balance and displays antimicrobial effects against pathogenic bacteria (Hara-Kudo et al., 2005). It has been shown that green tea polyphenols have strong antioxidation properties (Nishida et al., 2006). It reduces thiobarbituric acid reactive substances (TBARS) values and maintain oxodative stability of pig meat (Ko et al., 2008), broiler meat (Yang et al., 2003) and egg yolk (Uuganbayar et al., 2005). Yang et al. (2003) reported that cholesterol levels were decreased and fatty acids of plasma and meat were improved, when the animals were fed different levels of green tea by-products (GTB). Supplementation of GTB and green tea with probiotics has no negative effect on the blood components of beef cattle and calves (Lee, 2005; Sarker et al., 2010). Ko and Yang (2008) found that adding 0.5% GTB in the finishing diet produced positive effects on the humoral and cellmediated immunity of pigs. In the present study, we evaluated the activity of GTB to determine the specific effects of this component on growth performance, carcass characteristics, meat quality, blood parameters and immune response in finishing pigs and also to determine the effective level in finishing pig diet. MATERIALS AND METHODS Animals, diets and experimental design A total of 100 crossbreed (Landrace × Large White) castrated male finishing pigs with an average body weight of 77 ± 0.4 kg were assigned to 5 dietary treatments in a completely randomized design. Each treatment had 4 replications with 5 pigs per replication. The pigs were reared in the experimental farm at Sunchon National University following the guidelines for the care and use of animals in research (Korean Ministry for Food, Agriculture, Forestry and Fisheries, 2008). All animals were fed experimental diets for 6 weeks. The five dietary treatments were as follows: control (basal diet), antibiotic (basal diet with 0.003% chlortetracycline) and basal diet with 0.5, 1 and 2% GTB. Used green tea, residues from green tea drink manufacturing were collected from the green tea experimental station (Boseong, South Korea), dried, grounded and used for experiment. GTB was analyzed and the main constituents of were found as moisture 10.15%, crude protein 20.16%, crude fat 3.08%, crude fiber 19.20%, crude ash 5.22% and catechins 7.17%. All diets were formulated to meet or exceed nutrient requirements of finishing pigs (NRC, 1998). The formula and chemical composition Hossain et al. 2459 of the basal diet used in this experiment are provided in Table 1. Measurements and analysis Growth performance, carcass characteristics and oxidative stability of meat Body weights were measured on a biweekly basis from the beginning to the end of the experiment. Feed intake was determined by measuring feed residue on a biweekly basis from the start of the experiment. Feed conversion ratio (FCR) was obtained by dividing the feed intake by body weight gain. At the end of the experiment, pigs were transported to the slaughter house (Naju, Korea). Carcass weight and back fat thickness were measured, and carcass grade were determined according to Animal Products Grade System (Korea Institute for Animal Products Quality Evaluation, 2011). All the pork carcasses in South Korea are graded both in quality and conformation terms. The quality of pork carcasses is graded 1 + , 1 and 2 based on the marbling, lean color and conditions of belly streaks. The conformation terms of pork carcasses is graded A, B and C by assessing carcass weight, back fat thickness, balance, muscle, fat condition and so on. Carcass grades in this study were expressed as 3 (extremely good), 2 (good) and 1 (bad) in A, B and C grades of conformation terms. The thiobarbituric acid reactive substances (TBARS) value of pork was assayed using the methods described by Vernon et al. (1970) with slight modifications. Four gram of loin meat mixture was blended at full speed for 1.5 min with 10 ml of solution containing 20% trichloroacetic acid in 2 M phosphoric acid. The resulting sediment was transferred to 100 ml volumetric flask containing 8 ml distilled water and diluted by shaking and homogenized. The mixture was filtrated through Whatman No. 1 filter paper. Then 5 ml filtrate was transferred to a 20 ml test tube and 5 ml of 2-thiobarbituric acid (0.005 M in distilled water) was added. The solution was shaken in a water bath at 80°C for 30 min. After cooling, the color development was measured at 530 nm using a VIS- Spectrophotometer (Model 20D + , Milton Roy, PA, USA). Thiobarbituric acid reactive substances (TBARS) values were expressed as micromole of malondialdehyde per hundred gram of meat. Meat composition, quality and sensory evaluation Moisture, crude protein, crude fat, and ash percentage of the meat samples were analyzed according to Association of Analytical Communities (AOAC, 2000) methods. Color values on a freshly cut surface (3 cm thick slice) of loin meat were measured using a CR- 301 Chroma Meter (Minolta Co., Osaka, Japan) for International Commission on Illumination (CIE) standard of lightness (L), redness (a) and yellowness (b). Muscle pH values were determined using a pH meter (ATI 370, Orion Research Inc, MA, USA). The meat samples were broiled in a water bath at a temperature of 70°C for 30 min, surface dried, and weighed. Cooking loss was determined by expressing the cooked sample (B) weight as a percentage of the precooked sample (A) weight. Cooking loss (%) = [(A−B)/A] × 100. Water holding capacity was determined according to the procedure described by Wardlaw et al. (1973). Shear force was measured using the Instron Universal Testing Machine (Model 4465, Instron Corp., Norwood, MA, USA), which was equipped with a Warner- Bratzler shear device. From each cooked meat sample, a 0.5 × 4.0 cm (approximately 2.0 cm 2 ) cross section was cut for shear force measurements. The meat samples were placed at right angles to the blade. The crosshead speed was 100 mm/min and the full scale load was 50 kg. Sensor evaluation of pork meat was organoleptically evaluated by ten trained judges from the
2460 J. Med. Plants Res. Department of Animal Science and Technology at Sunchon National University on the three point hedonic scale for juiciness, tenderness and flavor. Biochemical and hematological parameters of blood Blood samples were collected from the jugular vein of the pigs by venipuncture on the last experimental day, 3 h after feeding. To analyze the biochemical composition of plasma, blood samples were separated by centrifuging for 20 min at 1500 (× g) and were total protein, cholesterol, glucose, albumin, globulin, albumin/globulin (A/G) and blood urea nitrogen (BUN) concentrations were measured using a blood analyzer (COBAS MIRA: Roche, Germany). Number of white blood cell (WBC, 10 3 /mm 3 ), red blood cell (RBC, 10 6 /mm 3 ) and hemoglobin concentration (g/dl) were determined using a hematological analyzer (XE-2100, Automated Hematology Analyzer, Sysmex America, Inc.) within 3 h after blood sampling. Spleen cells culture and proliferation The immune response of pig spleen cells was analyzed at the end of the experiment. At one third areas of the spleens of pigs, a 1 cm 3 tissue sample was extracted and split into single cell on bovine serum medium (RPMI-1640). Next, NycoPrep TM 1.077A was used to remove the dead cells and red blood cells. Spleen cells were cultured in RPMI 1640 medium supplemented with 10% (v/v) fetal calf serum, 2 mM l-glutamine, 100 units/ml penicillin and 100 μg/ml Table 1. Ingredients and chemical composition of the basal diet. Ingredients (%, as-fed basis) Amount Corn 44.65 Wheat 25.00 Wheat bran 4.00 Soybean meal 13.50 Lupin seeds 3.00 Limestone 0.77 Tricalcium phosphate 1.10 Salt 0.25 Vitamin-mineral premix 1 0.56 Animal fat 2.50 Molasses 4.50 L-lysine 0.17 Chemical composition (% dry matter) Crude protein 15.00 Calcium 0.78 Available phosphorus 0.55 Lysine 0.80 Methionine 0.27 Metabolisable energy (kcal/kg) 3160 1 Provided the following nutrients per kg of diet: vitamin A, 6000IU; vitamin D3, 800IU; vitamin E, 20IU; vitamin K3, 2 mg; thiamin, 2 mg; riboflavin, 4 mg; vitamin B6, 2 mg; vitamin B12, 1 mg; pantothenic acid, 11 mg; niacin, 10 mg; biotin, 0.02 mg; Cu, 21 mg; Fe, 100 mg; Zn, 60 mg; Mn, 90 mg; I, 1.0 mg; Co, 0.3 mg; Se, 0.3 mg. streptomycin, 1% (v/v) nonessential amino acids, and 0.05 mM 2mercaptoethanol (Gibco Paisley, UK) at 37°C, 5% CO 2 condition. Viable cells were counted in a hemocytometer by trypan blue exclusion.Triplicate cultures were performed in 96-well flatbottomed tissue culture plates (Costar, Cambridge, MA, USA) in a final volume of 200 µl per well containing 5 × 10 5 cells in the presence of different concentrations of LPS (lipopolysaccharide; 1.0, 3.0 and 10 μg/ml–Sigma-Aldrich, Saint Louis, MO, USA) or Con A (concanavalin A; 0.1, 0.3 and 1.0 μg/ml–Sigma-Aldrich, Saint Louis, MO, USA). Cultures were then incubated for 3 days at 37°C, 5% CO2. Cell proliferation was examined by MTS assay using the Celltiter 96 ® AQueous One solution Cell Proliferation Assay Kit (Promega, Madison, WI, USA) following the manufacturer’s instruction. The optical densities were measured in a microplate reader (Optimax, Molecular Devices, USA) at a wavelength of 490 nm. Absorbance was corrected by a prepared triplicate set of control wells (without cells) containing the same volumes of culture medium and CellTiter 96 ® AQueous One Solution Reagent, as in the experimental wells. We subtracted the average 490 nm absorbance, from the “no cell” control wells, from all other absorbance values to yield corrected absorbances. T cell subsets analysis Spleen lymphocytes (1 × 10 6 cells/ml) were stained with fluoresein isothiocyanate (FITC)-labeled anti-mouse CD3 and phycoerythrin (PE)-labeled anti-mouse CD4 + monoclonal antibodies for helper (CD3 + , CD4 + ) T cells, or fluoresein isothiocyanate (FITC)-labeled anti-mouse CD3 and fluoresein isothiocyanate (FITC)-labeled
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Journal of Medicinal Plants Researc
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Editors Prof. Akah Peter Achunike E
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Electronic submission of manuscript
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Fees and Charges: Authors are requi
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Table of Contents: Volume 6 Number
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Figure 1. Chuanxiong Rhizoma (http:
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Table 1. Identification of main pea
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Table 3. Pharmacokinetic parameters
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active component study, many invest
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and 1185 kg ha -1 in the first site
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Table 1. Recommendation for use fer
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Sadanandan AK, Hamza S (1996c). Stu
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and it has been traced to South Ame
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Table 3. Results of phytochemical a
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(a) (b) (c) Figure 1. (a) Normal ce
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emedies in the south of Brazil. J.
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known fatty acids and terpenoids we
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Table 1. Renal function tests of ra
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Al-Radahe et al. 2271 Figure 3.nnnn
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namely disruption of the vascular e
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Zayachkivska OS, Konturek SJ, Drozd
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β-carotene were purchased from Sig
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Figure 1. Effect of ethanol concent
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Table 3. Climatic and soil factors
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content of polysaccharides. Phospha
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Serge et al. 2285 Table 1. Relative
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Percentage inhibition % of inhibiti
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Table 1. Result of phytochemical sc
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Table 3. Result of thin layer chrom
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Table 2. MIC of a compound 1 from c
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Zirihi et al. 2301 Figure 2. Termin
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Figure 5. T. catappa Linn. (Combret
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Figure 8. A. fumigatus: Aspect of c
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Figure 12. Sensitivity of A. fumiga
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Noormi et al. 2311 Table 1. Callus
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Table 2. Contd. NoC=No callus. 4.0
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Figure 2. Contd. at 2.0 mg/L yellow
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Table 1. Biochemical parameters in
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Figure 3. Bcl-xL reactions in inter
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ACKNOWLEDGMENTS The authors would l
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Figure 1. The first page of the boo
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Figure 3. The most widely included
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Figure 4. Lithographic print “St.
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Table 1. List of plants included in
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Table 1. Contd. Nedelcheva 2333 Cin
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Table 1. Contd. Nedelcheva 2335 Rhe
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14 7 1 15 1 35 17 16 Imported plant
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Academy of Science Publishing House
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et al., 1996; van Wyk and Gericke,
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DPPH inhibition (%) DPPH % inhibiti
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% Mortality Concentration (µg/ml)
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information on plants used for the
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Table 1. Levels of independent vari
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Arbutin (mg/g) Co-solvent Figure 1.
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Arbutin (mg/g) Extraction pressure
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Table 3. Arbuitn content of Asian p
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Arbutin (mg/g) Extraction pressure
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Arbutin (mg/g) Extraction temperatu
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Table 1. Chemical composition of es
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Darjazi 2369 Table 2. Statistical a
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Table 3. Correlation matrix (number
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Table 1. Precision test. extraction
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Figure 2. Effect of different alcoh
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Table 6. Results of ANOVA. Nie et a
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Table 2. Plant data and MIC values
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exhibiting the polar extracts of P.
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Table 3. Contd. AcOEt ++ ++ ++ - Me
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Figure 1. Map of Ladakh region of I
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Warghat et al. 2391 Table 3. Summar
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Table 5. Inter-Population genetic i
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Excoffier L, Smouse PE, Quattro JM
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2524 J. Med. Plants Res. ACKNOWLEDG
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UPCOMING CONFERENCES 15th European
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