EVALUATION OF VITAMIN C AND POLYHERBAL ANTISTRESS SUPPLEMENTATION ON STRESS AND IMMUNE CHARACTERS IN BROILER CHICKEN

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2 Int. J. Agric.Sc & Vet.Med M Vijay Kumar et al., 2014 Research Paper ISSN Vol. 2, No. 4, November All Rights Reserved EVALUATION OF VITAMIN C AND POLYHERBAL ANTISTRESS SUPPLEMENTATION ON STRESS AND IMMUNE CHARACTERS IN BROILER CHICKEN M Vijay Kumar 1*, Shivi Miani 1,2 and Vinay Tikare 1 *Corresponding Author: M Vijay Kumar, vijaymatham@gmail.com A total of Ninety (90) day old broiler chickens were randomly divided into 3 groups and kept under elevated summer temperature to investigate the effect of ascorbic acid and AYUCEE on the antioxidative profiles viz., Thiobarbituric acid reacting substances (TBARS), Super oxide dismutase (SOD), Catalase (CAT) and Glutathione peroxidase (GSH) in both erythrocyte suspension and hepatic tissue concentrations. Heat stress was associated with an increase in TBARS and decrease in antioxidant enzyme profiles in untreated group while supplementation of Ascorbic acid (Vit. C) through feed and AYUCEE supplementation through drinking water has restored the depleted profiles of antioxidation enzymes of the birds thereby ameliorating heat stress and sustaining the productivity in the commercial broilers. Keywords: Ascorbic acid, Polyherbal antistressor formulation, Summer condition, Commercial broilers INTRODUCTION Poultry have the ability to synthesize ascorbic acid, but this ability is inadequate under stress conditions, such as high environmental temperatures, high humidity, a high productive rate and parasitic infestation. The impaired muscle membrane integrity in breast muscle of heatstressed broiler chickens was also considered to be related with the changed redox balance because broiler chickens that were exposed to acute heat stress exhibited more than a 2-fold increase of MDA as an indicator for lipid peroxidation, in the skeletal muscle (Mujahid et al., 2007). Several studies indicated that heat stress reduces the bodyweight (Al-Neemy and Hassan, 2002), immune response and also causes mortality and different therapeutic measures are used to minimize the harmful 1 Department of Veterinary Pharmacology and Toxicology, Veterinary College, Karnataka Veterinary Animal and Fisheries Sciences University, Post Box No. 6, Bidar , India. 2 Research and Development, Ayurvet Limited, Baddi, HP India. 104

3 effects of heat stress on performance of broiler chicks such as ascorbic acid, vitamin E (Sahin et al., 2003), acetylsalicylic acid (Naseem et al., 2005b), potassium chloride (Al-Khateeb and Al- Ddin Salih, 2005), sodium bicarbonate, acetic acid Hassan et al. (2009). The negative effects of high temperatures on poultry performance can be minimized by the use of appropriate housing design, installation of cooling systems, feed formulations designed according to feed intake and weather conditions, and the use of some minerals, electrolytes, ascorbic acid, or acetylsalicylic acid (ASA) in the drinking water of birds (Branton et al., 1986; Smith and Teeter, 1992; and Naseem et al., 2005a and 2005b). High ambient temperature negatively influences the performance of broilers. Biochemical and physiological changes associated with hyperthermia can potentially promote Reactive Oxygen Species (ROS) formation. This study was conducted to evaluate comparative effects of supplementation of ascorbic acid, and AYUCEE in diet and water on the performance of heat- exposed broilers. MATERIALS AND METHODS Experimental chicks: Ninety day-old unsexed broiler chicks of strain Vencobb were purchased from a local hatchery for this study and were randomly allotted to three treatment groups with each treatment having three replicates and each replicate having Ten birds. The brooder house as well as other equipment was thoroughly disinfected before the arrival of the chicks and maintained as per the Cobb Broiler Management Guide Cobb (2008). The birds were floor-brooded on wood-shaving in the experimental pens where they were allowed a week for adjustment. Additional source of heat was provided during the brooding period. Chicks were reared on deep litter, kept under 23 h light: 1 h dark and stocking density was 10 birds/m 2. During the first three weeks of trial the chicks were fed ad libitum with standard starter mash and thereafter with finisher mash. Different treatment groups were shown in the Table 1. The composition of the diets (starter and finisher phases) given to the birds is shown in Table 2. Water at ambient temperature was supplied ad libitum throughout the period of the experiment in plastic drinkers. Sacrifice Birds of all the groups were vaccinated with Marek s Disease (day 1) New castle Disease (ND) vaccine on 7 th and 21 st and 31 st day and Infectious Bursal Disease (IBD) vaccine on 14 th day. S. No. Treatment Group Table 1: Different Treatment Groups No. of Birds/Pen/ Replication No. of Replicates 1. Group-T 0 : Control (Environment stress without Antistressor No treatment 2. Group-T 1 : Summer stress Synthetic Vit. C (AA) 100 g/ (environmental) + synthetic ton of feed vitamin C 3. Group-T 2 : Summer stress Polyherbal antistressor + (environmental) formulation (Ayucee AYUCEE 100 g/ton of feed 105 Total Number of Birds/Treatment Dose (kg/tone Feed)

4 Table 2: Composition of Diets for Broiler (Starter and Finisher Phases) Ingredients Starter Phase (%) Finisher Phase (%) Maize Soybean meal Groundnut cake Fishmeal Wheat offal Bone meal Oyster shell Salt Premix Methionine Lysine Table 3: Nutritive Value of the Diet Calculated crude protein (%) M.E (Kcal/kg) Ether extract (%) Crude fibre (%) Calcium (%) Phosphorus (%) Note: *1kg of premix contains: Vitamin A-10,000,000 IU; Vitamin D3-2,000,000; Vitamin E-20,000 IU; Vitamin K-2,250 mg; Thiamine B1-1,750 mg; Riboflavin B2-5,000 mg; Pyridoxine B6-2,750 mg; Niacin- 27,500 mg; Vitamin B12-15 mg; Pantothenic acid-7,500 mg; Folic acid-7500 mg; Biotin-50 mg; Choline chloride-400 g; Antioxidant-125 g; Magnesium-80 g; Zinc-50 mg; Iron-20 g; Copper-5 g; Iodine-1.2 g; Selenium-200 mg; Cobalt-200 mg. The blood samples were drawn from the wing vein at 3 rd and 5 th week from the birds (identified by wing band numbers) in each group. RBC was collected by separating plasma from blood for estimation of Erythrocytic Lipid Peroxidation Profiles for erythrocytic antioxidant enzyme analysis, the erythrocyte pellet was washed three times in ice-cold NaCl (0.9%). Packed erythrocytes were resuspended in Phosphate Buffer Saline (PBS) and kept frozen at 20 C until used for the estimation of erythrocytic enzymes. The 1:10 dilution of erythrocyte suspension in PBS was used for the estimation of superoxide dismutase (SOD). The haemoglobin concentration in erythrocyte suspensions was determined by the cyanmethemoglobin method. The birds were sacrificed at the end of 3 rd and 5 th week and hepatic tissues were collected for the assay of antioxidant Profiles viz., Thiobarbituric Acid Reacting Substances (TBARS) (Balasubramanian et al., 1988) Superoxide dismutase (Madesh and Balasubramanian, 1998) Catalase: (Caliborne, 1985), reduced glutathione (GSH) (Moron et al., 1979) and Protein carbonyls (Levine et al., 1990). Statistical Design and Analyses The data were statistically analyzed by applying paired sample t-test. Remaining data were subjected to statistical analysis by applying one way ANOVA using Statistical Package for Social Sciences (SPSS) 15.0 version. Differences between means were tested using Duncan s multiple comparison test and significance level was set at RESULTS AND DISCUSSION The stress of high ambient temperature may negatively influence the performance of broiler chickens by reducing feed intake, live weight gain and feed efficiency (Donkoh, 1989). Several methods are available to alleviate the negative effects of high environmental temperature on performance of poultry. But of the high cost and impracticality of cooling animal buildings, interest on dietary manipulations has increased Njoku (1986). LPO level is a direct indication of oxidative damage of cells as seen in aging, atherosclerosis 106

5 and other pathological disorders. LPO of serum lipids, especially those of High Density Lipoproteins (HDL) is highly unfavorable as it may end up with many vascular disorders. Results of the present study revealed that, LPO levels in serum and liver of HST-treated birds were significantly less than those encountered in HST-untreated birds. These results suggestive of beneficial aspects of supplementation of AYUCEE on par with Synthetic Vit. C continuously for 42 days under HST and continued during the period of HST in birds; thereby undesired side effects produced by free radicals that were generated could be compromised successfully. GPX and GR are enzymes associated with conversion of GSH to oxidized form. Liver is the chief organ concerned with detoxification process and availability of increased amount of GSH in the liver improves this function. Results of the present study revealed that the GSH content of liver of HST treated birds was elevated concomitant with the activities of GPX and GR enzymes significantly over control group. SOD enzyme has also a minor role to play in oxidizing GSH to GSSG. Liver SOD activity was increased in HST treated birds when compared with untreated birds because increased formation of GSH triggered the oxidation reactions mediated by GPX and SOD. However there were no such appreciable changes in CAT and SOD activities in liver tissue of HST-untreated birds. Moreover there was an insignificant increase in the activity of liver enzyme such as GPX and a great deal of decreased activity of GR in HST-untreated birds that resulted in the accumulation of GSH without being oxidized. In other words, the rate of forming of GSH and the rate at which it was oxidised by antioxidant liver enzymes did not match in HSTuntreated birds while in HST treated birds an appreciable balance between activities of oxidising and reducing enzymes of glutathione could be observed. CONCLUSION Because of increased activity and concentration of enzymatic and non-enzymatic antioxidants resulted by supplementation of AYUCEE water, birds could remained healthy, maintained body weight and were acclimated to the heat stressor quicker than untreated groups. In other words AYUCEE treatment facilitates adaptation of the chicken to stressful hot conditions at an early date on par with synthetic Vitamin C. REFERENCES 1. Al-Khateeb F S and Al-DdinSalih S (2005), Effect of Withdrawal and Supplementation Potassium Chloride to Reduce the Negative Impact of Heat Stress on the Productive and Physiological Performance of Broiler, Ms.C. Thesis, Animal Resources Dept., University of Mosul. 2. Al-Neemy D T and Hassan Y (2002), The Effect of Withdrawal, Supplementation Potassium Chloride and Acclimation to Alleviate the Heat Stress in Some Physiological Characteristics and Productive Performance of Broiler, Ph.D Thesis, Animal Resources Dept., University of Mosul. 3. Balasubramanian K A, Manohar M and Mathan V I (1988), An Unidentified Inhibitor of Lipid Peroxidation in Intestinal Mucosa, Biochim. Biophysic. Acta, Vol. 962, pp Branton S L, Reece F N and Deaton J W (1986), Use of Ammonium Chloride and Sodium Bicarbonate in Acute Heat Exposure of Broilers, Poult. Sci., Vol. 65, pp

6 5. Caliborne A (1985), Catalase Activity, in Greenwald R A (Ed.), CRC, Handbook of Methods of Oxygen Radicals Research, pp , CRC Press, Boca Raton, FL. 6. Cobb-Vantress (2008), Broiler Management Guide. 7. Cook G C and Sherlock S (1965), Results of a Controlled Clinical Trial of Glutathione in Cases of Hepatic Cirrhosis, Gut., Vol. 6, pp Donkoh A (1989), Ambient Temperature: A Factor Affecting Performance and Physiological Response of Broiler Chickens, Int. J. Biometeoro., Vol. 33, pp Hassan A M H, AbdelAzeem M and Reddy P G (2009), Effect of Some Water Supplements on the Performance and Immune System of Chronically Heat- Stressed Broiler Chicks, Int. J. Poult. Sci., Vol. 8, pp Levine R L, Garland D, Oliver C N, Amici A, Climent I, Lenz A G, Ahn B W, Shaltiel S and Stadtman E R (1990), Determination of Carbonyl Content in Oxidatively Modified Proteins, Methods Enzymol., Vol. 186, pp Madesh M and Balasubramanian K A (1998), Microtiter Plate Assay for Superoxide Dismutase Using MTT Reduction by Superoxide, Indian J. Biochem. Biophys., Vol. 35, No. 3, pp Mujahid A, Pumford N R, Bottje W, Nakagawa K, Miyazawa T, Akiba Y and Toyomizu M (2007), Mitochondrial Oxidative Damage in Chicken Skeletal Muscle Induced by Acute Heat Stress, Poult. Sci., Vol. 44, pp Naseem M T, Naseem S, Younus M, Zafar I C, Aamir G, Asim A and Akhter S (2005a), Effect of Potassium Chloride and Sodium Bicarbonate Supplementation on Thermotolerance of Broilers Exposed to Heat Stress, Int. J. Poult. Sci., Vol. 4, pp Naseem S, Younus M, Bilal A, Aamir G, Asim A and Akhter S (2005b), Effect of Ascorbic Acid and Acetylsalicylic Acid Supplementation on Performance of Broiler Chicks Exposed to Heat Stress, Int. J. Poult. Sci., Vol. 4, pp Njoku P C (1986), Effect of Dietary Ascorbic Acid (Vitamin C) Supplementation on the Performance of Broiler Chickens in a Tropical Environment, Anim. Feed Sci. Technol., Vol. 16, pp Sahin K, Sahin N and Kucuk O (2003), Effects of Chromium and Ascorbic Acid Supplementation on Growth, Carcass Traits, Serum Metabolites and Antioxidant Status of Broiler Chickens Reared at a High Ambient Temperature (32 C), Nutr. Res., Vol. 23, pp Moron M S, Depierre J W and Mannervik B (1979), Levels of Glutathione, Glutathione Reductase and Glutathione S-Transferase Activities in Rat Lung and Liver, Biochim. Biophys. Acta, Vol. 528, pp

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