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ThesisItem Open Access Cell mediated immune response in cockerels under temperature stress(Department of Veterinary Physiology, College of Veterinary and Animal Sciences, Mannuthy, 2008) Savitha, M K; KAU; Ramnath, VA study was conducted with the objective of finding the variations in the cell mediated immunological responses induced by heat / cold stress in cockerels and the role of brahma rasayana (BR) supplementation as an immunopotentiative agent. A total of 144 Gramapriya cockerels having 1 kg bw (4 months) were subjected to heat (40 ± 10C and relative humidity (RH) 80 ± 5 per cent) and cold (4 ±10C and RH 45 ± 5 per cent ) each for 4 consecutive hours per day for 10 days in a controlled environmental chamber (floor space 875 cm2 / bird). Control groups were reared randomly under ambient temperature of 30 ± 10C and RH 65 per cent. The dietary inclusion of BR was done @ 2.0 g/kg bw for 20 days (10 days prior to and during the period of heat / cold stress). To certain extend the heat stress induced hyperthermia in birds. Heat stress was positively correlated with the plasma corticosterone level in untreated cockerels, whereas exposure to cold and BR treatment resulted in lowered plasma corticosterone concentration. The phagocytic potential and migratory capability of peritoneal macrophages was found to be enhanced in both heat / cold stress, which were further enhanced by BR supplementation, while the cytotoxicity against L929 cells was decreased in heat / cold stressed cockerels, which were reversed by BR supplementation. It was noticed that the cutaneous delayed hypersensitivity response in heat / cold stressed untreated cockerels at 24 / 48 h post PHA inoculation got increased by BR supplementation. Similarly, the leucocyte migration inhibition index which was decreased in temperature stress could be reversed by BR treatment. Administration of BR was found to enhance the proliferation of splenocytes in response to mitogens like PHA and Con A in both heat / cold stress. The mitogenic response of PHA was dose dependent, while that of Con A was not dose specific. The natural killer cell activity was found decreased by heat / cold stress and enhanced with the supplementation of BR, earlier lysis of target K562 cells were achieved by effector cells. The antibody dependent cellular cytotoxicity was enhanced in heat / cold stressed cockerels administered with BR on the ninth day, when compared to their untreated counterparts, which was measured from the percentage SRBC lysis. An earlier antibody dependent complement mediated cytotoxicity was observed on 5th day of the experiment, while the BR supplementation could augment the tumoricidal property through ACC in heat / cold stressed and non-stressed cockerels. The cockerels under induced heat stress exhibited much diminished cell mediated immune profile when compared to cold stressed and non-stressed birds. In general, most of the immune parameters screened in cockerels under induced cold stress were found to be in par with non-stressed BR treated group. This indicated that there was not much alteration in the immune status of cockerels during cold exposure. Results of the present study also indicated that the dietary supplementation of BR @ 2.0 g/kg bw in poultry could potentiate the immunological insult inflicted by heat stress.ThesisItem Open Access Physio-biochemical evaluation of broiler chicken fed with processed fish wastes(Department of Veterinary Physiology, College of Veterinary and Animal Sciences, Mannuthy, 2008) Darsana, M G; KAU; Sreekumar, K PThe present study was undertaken to study the effect of dietary incorporation of processed fish wastes; acid silage (fish waste) and surimi waste powder on the physio-biochemical parameters in broiler chicken. The experiment was conducted on forty five, day old broiler chicken of Vencob strain for a period of seven weeks. After three weeks of age, they were randomly divided into three groups G-I, G-II and G-III of 15 birds each. During the first three weeks, they were fed with standard broiler starter ration of BIS specification. After three weeks, G-I was fed with the standard broiler finisher ration, G-II was fed with the standard broiler finisher ration in which there was 100 per cent replacement of unsalted dried fish with acid silage (fish waste) and G-III was fed with the standard broiler finisher ration in which there was 100 per cent replacement of unsalted dried fish with surimi waste powder. The rations of G-I, G-II and G-III were made isocaloric and isonitrogenous. Body weight and feed consumption were recorded at weekly intervals from fourth to seventh weeks of age. Blood samples were collected fourth to seventh week from the three groups at weekly intervals and haematological and biochemical parameters were analysed. Haematological parameters like Hb, VPRC, TEC, TLC, serum protein profile like total protein, albumin, globulin, albumin globulin ratio, serum lipid profile like total lipids, triglycerides, HDL,VLDL, NEFA, total cholesterol, serum enzymes like ALT,AST antioxidants like blood catalase, blood SOD, serum GSH, serum LPO, serum electrolytes like Na, K, Ca, Mg, Fe were estimated. At the end of the experiment, the birds were sacrificed and giblet weight was assessed. The meat of three representative samples from each group was used for analysing meat cholesterol and the proximate principles like crude protein, ether extract and total ash. There was no significant difference between the three groups in body weight and feed consumption and they were gradually increasing during the experimental period in all the three groups. The haematological parameters also were comparable between the control and experimental groups. No significant difference was observed between the groups in the concentrations of total protein, albumin and albumin globulin ratio throughout the study. The concentration of globulin was significantly higher in the control group at the start of the study but later, the values were significantly comparable between the three groups. The concentrations of total lipids, triglycerides, NEFA, VLDL and total cholesterol maintained a similar trend between the control and experimental groups but at the seventh week, the concentration of HDL in the acid silage (fish waste) fed group was lower than that of the control group. The concentrations of serum enzymes and minerals (Na, K, Ca and Fe) did not show any significant change between the three groups during the entire study. Though the Mg level showed a fluctuating trend at fifth and sixth weeks of age, at the end of the study, there was no significant difference between any of the groups. The antioxidant status was also significantly comparable between the groups. The meat parameters like giblet weight, meat cholesterol and the proximate principles were also significantly similar in the three groups. It is obligatory to use animal protein source in broiler chicken for obtaining better growth performance. Fish meal is rich in certain amino acids essential for the proper growth of chicken that are limiting in the vegetable protein supplements. Unconventional protein sources like fish wastes are to be used in the ration due to the high cost and low availability of fish meal. In the present study, physio- biochemical effects of using processed fish wastes in the ration of broiler chicken instead of fish meal were assessed and they induced a growth similar to that of the standard normal ration which contain fish meal as animal protein. Thus the incorporation of the two fish wastes could be advocated to the farmers to reduce the feed cost and improve the profit without affecting the growth and meat quality of broiler chicken and also as a measure to minimize environmental pollution.ThesisItem Open Access Evaluation of antistress and growth promoting effect of aswagandha (Withania somnifera) in broiler chicken(Department of Veterinary Physiology, College of Veterinary and Animal Sciences, Mannuthy, 2009) Rekha P, Raghavan; KAU; Sreekumar, K PThe purpose of the study was to asses the antistress and growth promoting effect of Withania somnifera in broiler chicken. Sixty numbers of day old broiler chicks (vencob) procured from a commercial hatchery were reared under standard managemental conditions. At day old they were randomly divided into six groups (G1 to G6) with ten birds in each group. The study was conducted upto sixth week of age. The birds in various groups were as follows, G1- Normal , G2- Normal +5 g W. somnifera per kg feed, G3- Normal + 10 g W. somnifera per kg feed, G4- Stressed, G5- Stressed + 5 g W. somnifera per kg feed, and G6- Stressed + 10 g W. somnifera per kg feed. Production parameters such as body weight, weekly weight gain, feed consumption and feed efficiency were evaluated on weekly basis. All birds were immunized with 1ml of 7 per cent sheep red blood cells intravenously five days before subjecting to stress, for the evaluation of immune status. Stress was induced from four weeks to six weeks of age by reducing the floor space for a bird 1/3rd (372 cm2/ bird) of the optimum requirement (1116 cm2/bird). Blood samples were collected from wing vein, five days and one day before the induction of stress as well on day one, three, six, nine, twelve and fifteen days after the induction of stress. Haematological parameters such as total erythrocyte count (TEC), total leukocyte count (TLC), haemoglobin (Hb), heterophil lymphocyte ratio (H:L), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC) and volume of packed red cells (VPRC) were determined. Serum was utilized for analysis of total protein, albumin, globulin, glucose, C-reactive protein (CRP), alanine amino transferase (ALT) and total cholesterol. The A:G was derived. The immunological status was studied by haemagglutinin assay (HA). Tissues from liver were used for estimating tissue lipid peroxidation (LPO) and reduced glutathione (GSH) levels. The mucous membrane of proventriculus and duodenum were used for protease and amylase estimation respectively. The whole pancreas was used for pancreatic lipase estimation. Slaughter weights, carcass weight, giblet weight, weight of organs (spleen, bursa and adrenal) were also studied. The result of the present study revealed that supplementation W. somnifera did not affect the normal growth of broiler chicken. There was not much variation in slaughter weight, carcass weight and weight of organs between W. somnifera supplemented and not supplemented group. In stressed birds supplementation of W. somnifera ameliorated the stress related reduction in organ weight. Supplementation of W. somnifera did not affect the haematological parameters such as Hb, VPRC, TEC, TLC, MCV, MCH, MCHC and H:L in normal birds. Supplementation of W. somnifera to stressed birds reduced the stress induced increased TEC, MCV and H:L in birds. The supplementation of W. somnifera did not affect the serum total protein, albumin, globulin and A:G in normal birds. But in stressed birds the increased total protein was decreased by supplementation of W. somnifera. The supplementation of W. somnifera did not cause a sustained increase in ALT values revealing that it was not hepatotoxic to broilers. Supplementation of W. somnifera to stressed birds reduced the stress related hyperglycemia and hypercholesterolemia. The increased level of C-reactive protein in stressed birds was reduced through supplementation of W. somnifera. Supplementation of W. somnifera increased the GSH and decreased LPO levels in birds. Supplementation of W. somnifera ameliorated the stress related oxidative damage through the reduction of LPO and increase of GSH levels. W. somnifera improved the immune status of normal birds as well as that of immunosuppressed stressed birds. Besides, it also improved the activities of gastrointestinal enzymes in normal and stressed birds.ThesisItem Open Access Dietary supplimentation of spirulina platensis on growth and immune response of broiler chicken(Department of Veterinary Physiology, College of Veterinary and Animal Sciences, Mannuthy, 2008) Preetha, K; KAU; Girish Varma, GThe present experiment was aimed at investigating growth promoting and immunomodulating effects of Spirulina platensis in broiler chicken. Thirty two numbers of day-old broiler (Vencob strain) chicks were procured and reared in battery cages under standard management conditions up to sixth week of age. The birds were randomly divided into four groups, viz. GI, GII, GIII and GIV, having eight birds in each group. Birds in groups GI (control) were fed with standard broiler starter ration (from first to fourth week) and standard finisher ration (from fifth to sixth week). The experimental groups GII, GIII and GIV were fed with standard rations supplemented with Spirulina platensis at the rate of 0.50 per cent, 1.50 per cent and 3.00 per cent, respectively. Both control and experimental rations were made isocaloric and isonitrogenous. Birds were provided with ad libitum feed and clean drinking water throughout the experiment. Bodyweight of individual birds were recorded on day one, eight, 15, 22, 29, 36 and 43. Blood was collected at fourth, fifth and sixth week of age and whole blood/plasma was used for further estimations. Estimation of total erythrocyte count (TEC), blood haemoglobin concentration (Hb), volume of packed red cells (VPRC), total leukocyte count (TLC), differential leukocyte count (DLC), and blood glucose was made using whole blood samples. Blood plasma was utilised for estimation of concentrations of total protein, albumin, globulin, total lipid, total cholesterol, triglyceride, HDL cholesterol, LDL cholesterol, and VLDL cholesterol. Erythrocytic indices (MCV, MCH and MCHC), heterophil: lymphocyte (H:L) ratio and albumin: globulin (A:G) ratio were calculated from the respective values. The data was analysed by Split plot analysis (Split on time) as suggested by Rangaswamy (1995). Bodyweight of Spirulina-fed birds (GII, GIII and GIV) were significantly (P≤0.01) higher than that of control group throughout the experiment in an age and dose dependent way. Significant (P≤0.01) age and dose dependent increase in TEC, Hb, VPRC, MCH and MCHC and decrease in MCV values were observed in Spirulina-fed groups, suggesting strong erythropoietic effect of Spirulina. Significant (P≤0.01) age and dose dependent increase in total leukocyte and lymphocyte counts and decrease in heterophil, eosinophil, and monocyte counts were noticed in Spirulina-fed groups. Total leukocyte count was higher in Spirulina-fed birds, but without increase in heterophil count, ruling out the possibility of inflammatory responses. Additionally, reduced stress and more vigilant humoral immunity could be the implication. Also, Spirulina induced significantly (P≤ 0.01) more plasma albumin and plasma globulin, and thus, total plasma protein output in the recipients. Since the albumin: globulin (A:G) ratio did not change significantly, both these classes of proteins were equally promoted. This would mean a general boost up in plasma protein output by the liver as well as more immunoglobulin production by the lymphocytes. Increased number of lymphocytes in Spirulina-fed birds could be attributed to the higher plane of globulins in this experiment. Plasma lipid profile showed age and dose dependent lowering of total lipid, cholesterol, triglyceride, LDL and VLDL cholesterol in Spirulina-fed groups compared to control group, indicating that Spirulina would reduce lipid level, especially the ‘undesirable’ fractions, signifying the hypolipidaemic / lipolytic activity of the alga. The significant (P≤0.01) hypoglycaemic effect of Spirulina supplementation in the present study may indicate an anti-diabetic effect, which may indicate, in turn, propensity to have more lean meat than fatty meat in the subjects. The growth-promoting and erythropoietic effects of Spirulina platensis in broiler chicken may indicate its effectiveness as a nutritional supplement, while the hypolipidaemic and hypoglycaemic effects may imply better consumer safety and desirability to the broiler meat, which inturn, can potentially fetch premium price to the produce in the market.