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ThesisItem Open Access LEPTIN RECEPTOR EXPRESSION IN CULTURED OVARIAN FOLLICLES OF SHEEP(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-05) SRAVANI PRAGNA, K; Siva Kumar, A.V.N(MAJOR); Rambabu Naik, B; Punya Kumari, BABSTRACT : The expression of Leptin receptor (LepR) mRNA and protein was studied in sheep using Real time quantitative PCR and Immunohistochemistry in the cumulus cells and oocytes from : (i) In vivo grown preantral, early antral, antral, large antral follicles and Cumulus Oocyte Complexes (COCs) obtained from large antral follicles and subjected to 24h of In Vitro Maturation ( IVM) and (ii) Preantral follicles (PFs’) exposed to TCM199B, TCM199BL (10ng/mL) and TCM199BHGL (10ng/mL) media for 3min, cultured in vitro for two, four or six days and subsequently matured in vitro for 24h in respective cultures separately. LepR was observed at all stages of in vivo grown and in vitro cultured ovarian follicles in both cumulus cells and oocytes. Expression levels and patterns of LepR mRNA in PFs’ cultured in TCM199BHGL was similar to in vivo in all the stages except in cumulus cells from COCs after in vitro maturation for 24h, where the expression was significantly higher which was a positive effect. The expression of LepR in cumulus cells among the three different in vitro culturing conditions was significantly higher in PFs’ cultured in TCM199BHGL medium. In the oocytes, the Leptin receptor expression was similar or significantly higher in PFs’ cultured in TCM199BHGL medium than in vivo grown follicles except in PFs’ exposed to 3min stage which could suggest the synergistic action of growth factors and hormones with Leptin. Immunohistochemistry studies revealed highest intensity of Leptin receptor protein expression in the oocytes. Mild to moderate staining was observed in cumulus cells with good intensity in large antral stages. This protein expression coincided with gene expression. From our studies it is concluded that Leptin along with other growth factors and hormones supplementation stimulated the expression of Leptin receptor mRNA and protein on par with that of in vivo /even higher than that of in vivo condition.ThesisItem Open Access INFLUENCE OF LEPTIN ON THE EXPRESSION OF p53 and BAK GENES IN CULTURED OVARIAN FOLLICLES IN SHEEP(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2015-11) PRABHAVATHI, T; RAO, V.H(MAJOR); SIVA KUMAR, A.V.N; VEERABRAHMAIAH, KABSTRACT : The present study was conducted to know the influence of Leptin on the expression of p53 and BAK (apoptotic) genes in the sheep preantral follicles (PFs’) cultured in vitro. Ovaries of sheep were collected from the local slaughter house. Intact PFs’ (250-400μm) were isolated and placed individually in 20μl droplets of standard medium (TCM 199 B + 2 μg/ml FSH, 0.1μg/ml T4 + 10 ng/ml IGF-I + 1 mIU/ml of GH) supplemented with Leptin (10ng/ml) for 2, 4 or 6 days. Quantitative expression of Test genes (p53 and BAK) and Reference genes (18s RNA, RPLPO, HPRT1) were studied in the cumulus cells and oocytes isolated at different developmental stages (preantral, early antral, antral and large antral follicles) of the in vivo and in vitro grown follicles. The entire experiment was repeated twice. Duplicate samples of cDNA from each replicate of the experiment were subjected to relative-RT-qPCR. The pattern of expression of p53 gene in the cumulus cells from ovarian follicles grown in vivo and cultured in Leptin was similar from preantral to large antral follicle stage. In the oocytes, however, such similarity was restricted up to the antral follicle stage only. It is concluded that Leptin in culture medium mimiced the pattern of p53 expression in in vitro as in in vivo to some extent. The BAK gene expression was undetected in the cumulus cells and oocytes isolated from all the stages of both in vivo and in vitro developed ovarian follicles. Accordingly it is concluded that (i) Leptin has no influence on the expression of BAK gene in cultured ovarian follicles in sheep (ii) BAK may not be an important regulator of apoptosis in ovarian follicles and (iii) p53 plays a relatively more significant role than BAK in the regulation of apoptosis in ovarian follicles.ThesisItem Open Access AMELIORATION OF HEAT STRESS INDUCED OXIDATIVE DISTURBANCES BY HERBAL ANTIOXIDANTS IN BROILER CHICKEN(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2011-04) SWATHI, B; Gupta, P.S.P(MAJOR); Nagalaxmi, D; Rajasekhar Reddy, A; RaJu, M.V.L.NABSTRACT: A study was petformed to evaluate the antioxidative potential of herbals Tulsi (Ocimum sanctum) and Turmeric (Curcuma longa) in combating heat stress with two batches of broiler chicks in hot summer months. Each batch consisted of a total of 108 one day OM Vencobb broiler chicks which were grouped and offered 9 types of dietary regimens wtth variable concentrations and combinations of antioxidants. A group of 12 chicks was raised separately in stress free environment on basal diet without any antioxidant supplementation as control. Body weights and feed intake were measured at weekly intentab. The blood samples were colleded at 4" and 6" wk of age and haernatological estimations (RBC, WBC, Hb. & PCV) were carried out within 2 hrs of collection. While serum samples were preserved at -20°C until utilized for estimation of protein profiles (TP, albumen, globulin concentrations and A:G ratios), serum cortisol concentrations, humoral and cell mediated immune responses. Plasma samples were prepared by centrifugation, stored and utilized for analysis of enzymatic and non enzymatic antioxidant concentration. At the end of trail, breast and meat samples were procured by excision of sacrificed birds for studying organoleptic properties and lipid peroxidation. Histopathological studies of liver, bursa, brain and spleen were conducted to observe the tissue changes at the end of the experiment. The performance parameters (Body weights and feed efficiency), haernatologoical parameters (RBC, Hb. PCV and WBC), enzymatic (GSH-PX, Catalase and SOD), non enzymatic (Reducedglutathione) antioxidant concentrations, immunological indices (HI titers and CMI responses) were signifiintly low (PsO.01) in heat stressed birds compared to control and antioxidant supplemented groups. However, WBC count, serum cortisol concentrations, serum ALP and ALT activities were significantly (PS0.01) higher in heat stressed birds compared to control group. Histopathological study evidenced degenerative changes and pathological lesions in tissues collected from heat 8tmssed birds, While no lesions of pathological importance were observed with the tissues d control group. Sensory chamderistics of breast meat samples from heat stressed birds scored low on 5 point hedonic scale (3.42 for colour, 3.12 for texture, 3.42 for juiciness, 3.22 for tlavour and 3.14 for overall acceptability) compared to higher scores recorded for meat samples of control group. Dietary supplementation with herbals Tulsi and Turmeric was proved beneficial in terms of broiler performance, haematological, serological values. immunological indims and antioxidant status. Their inclusion also contributed to low levels of serum cortisol and improved sensory characteristics and reduced lipoperoxidative damage of broiler meat. Of the two herbals employed for the study. Turmeric had contributed to hgher body weights (1629.17 g and 1627.50 g with 0.2% and 0.4% level of inclusions, respectively), which were comparable with the weights achieved by supplementing the diet either with vitamin E alone (1637.52 g) and its combination with Selenium (1656.67 g) end control group (1684.17 g) at the end of 6'" week, Feed efficiency though low in the initial stages, an accelerated trend was noticed from 4m wk onwards wrth Turmeric supplementation. And even sensory attributes showed higher sensory scores with Turmeric supplementation at either of the dose. While, Tulsi, contributed to significantly (Ps0.01) higher concentration of enzymatic and non enzymatic antioxidants, blood and serum variables, and immunological indices at either of the dose supplemented. Herbals at higher concentration (Ocimum at 0.5% and Turmeric at 0.4%) had shown improved antioxidant -8. However, the two herbals when given in combination at different doses could not yield any additive effect. The benefdal results obtained with the supplementation of herbals at different doses were however remained low when compared to the values obtained with the diets inclusive of vitamin E alone, or its combination with Se and control group. But, considering the cost of these synthetic antioxidants vitamin E and Se and maintaining the birds in stress free environment, the herbals employed Tulsi and Turmeric in this study may be suggested as natural and economical alternatives in amelioration of heat stress and achieving higher performanceThesisItem Open Access EFFECT OF PROBIOTICS ON PERFORMANCE, LIPID PROFILE AND IMMUNE STATUS IN BROILER CHICKEN(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2010-12) VINAYA SREE, C; GUPTA, P.S.P(MAJOR); KONDAL REDDY, K; Gopala Reddy, A; Nagalakshmi, DABSTRACT: The effect of feeding probiotics on growth performance, blood constituents and immune parameters in broiler chicks was studied. An experiment of 42 days duration was conducted 525 day-old broiler chicks, which were randomly allotted to 7 dietary treatments. Each treatment contained 75 chicks. The 7 dietary treatments were control (no probiotic) and other 6 diets were the control diet supplemented with various probiotics viz., Pediococcus acidilactici (MTCC, Chandigarh), Pediococcus acidilactici (NCIM, Pune), Saccharomyces cerevisiae, Saccharomyces boulardii 50 % + Pediococcus acidilactici (NDRI) 50%, Saccharomyces boulardii, Lactobacillus reuteri @ of 109 CFU/gm. Data was collected for body weight, daily weight gain, feed intake and feed conversion ratio during the experimental period (0 – 42 d of age). On day 28 and 42, concentration of proteins, albumin, globulin, albumin/globulin ratio, cholesterol and triglycerides in serum, red blood cells (RBC), haemoglobin, packed cell volume (PCV) and white blood cells (WBC) in chick’s blood were evaluated. Body weight gain of broilers was influenced by inclusion of probiotics during over all period and groups supplemented with Pediococcus acidilactici, (MTCC, Chandigarh), Saccharomyces cerevisiae and combination of Saccharomyces boulardii 50 % and Pediococcus acidilactici 50% recorded the highest body weight amongst the experimental groups at the end of 6th week. Supplementation of probiotics as single or as their combination did influence feed intake in broilers, but had significant effect on feed conversion ratio of broilers for over all period. Livability was not influenced by various probiotics supplementation. The concentration of total proteins and globulins were significantly (P<0.05) higher in all the probiotic supplemented groups and Lactobacillus reuteri recorded the highest protein and globulin concentration. A/G ratio were significantly (P<0.05) lower in all groups while albumin concentration did not reveal any significant changes among the groups. Supplementation of probiotics had profound effect on the serum lipid profile, both serum cholesterol and triglycerides level was significantly (P<0.05) lowered with either of the probiotic supplementation. Mean TEC (106/cmm) was significantly (P < 0.05) lower in control group in comparison to other groups supplemented with probiotics and highest TEC was found in groups Saccharomyces cerevisiae, Lactobacillus reuteri and Saccharomyces boulardii. The mean mean TLC (103/cmm) was highest with Pediococcus acidilactici (MTCC, Chandigarh), Pediococcus acidilactici, (NCIM, Pune) and Saccharomyces cerevisiae. The probiotics Saccharomyces cerevisiae and Saccharomyces boulardii revealed highest Hb (g/dl) at the end of experimental period. The PCV (%) showed a significant (P < 0.05) decrease in control in comparison with that of probiotic fed groups at the end of 6th week and highest PCV (%) was observed with Saccharomyces cerevisiae and Saccharomyces boulardii. The humoral immune response to new castle disease virus at 28 days of age significantly (P<0.05) increased with the inclusion of probiotics. At the age of 42 days, HI titers were significantly (P<0.05) higher in all diets supplemented with probiotic than control. Supplementation of probiotic influenced the faecal pH which significantly (P<0.05) reduced in all dietary treatment groups compared to the control diet. Supplementation of probiotics in all dietary treatment groups lowered the faecal coliform count. The present study indicated that supplementation of probiotics @ (1×109 cfu/g) in diets of broiler diets improved growth, lowered serum lipid profile, increased serum protein profile, haematological constituents, faecal pH and faecal coliform count and better immune responses.ThesisItem Open Access In Vitro Culture Of Preantral Follicles From Goat Ovaries(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2002-01) Rajarajan, K; Rao, V.H(MAJOR); Amarnath, D; Haragopal, VThesisItem Open Access Different Response Of Groundnut (Arachis Hypogeae L.)Genotypes To Aluminium Toxicity(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2001-08) Prathap, Sunkara; Balakrishna Reddy, K(MAJOR); Eswara Reddy, N.P; Surya Prakasa Rao, J; Srinivasulu Reddy, KThesisItem Open Access Cryopreservation Of Rabbit Embryos By Open Pulled Straw (Ops) Method(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2001-06) Rambabu Naik, B; Rao, V.H(MAJOR); Amarnath, D; Gnanprakash, MThesisItem Open Access In Vitro culture Of Preantral Follicies Of Sheep(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2001-06) Tamilmani, G; Rao, V.H(MAJOR); Amarnath, D; Mutha Rao, MThesisItem Open Access IN Bitro Culture Of preantral Follicles From Sheep Ovaries(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2000-02) Hema Malini, N.C.; Somasekhar Reddy, K(MAJOR); Seshagir Rao, A; Kalakumar, B