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Sri Venkateswara Veterinary University, Tirupati

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  • ThesisItemOpen Access
    HEMATOBIOCHEMICAL PROFILE AND ELECTROPHORETIC PATTERN OF SERUM PROTEINS IN PUNGANUR CATTLE DURING DIFFERENT SEASONS
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2021-07) BUJJI, BANDARU; ASWANI KUMAR, K (MAJOR); Prasada Rao, T; Srinivasa Prasad, CH
    A total of sixty apparently healthy Punganur cattle aged two years and above grouped as: Females (n=30) and Males (n=30) maintained at Livestock Research Station, SVVU, Palamaner, Chittor district were selected to study hemato-biochemical profiles and to compare the electrophoretic pattern of serum proteins over different seasons.THI findings during the study period suggests that the summer season was having moderate heat stress (79.03±1.22) on Punganur cattle. Hematological parameters such as Hb, TEC, PCV, MCV, MCH, MCHC, Monocytes, Granulocytes, Lymphocytes and Platelets did not show any significant change among all groups between seasons. WBC values were high during summer in males and herd while there was no change in females. Total Protein and globulins were high during summer while albumins and A/G ratio showed higher values during winter in all the groups. BUN and Creatinine concentration were high in summer compared to other seasons, but the values were within normal range. Blood glucose concentration was low in summer in all the groups while there was no change in Total Bilirubin. Total Cholesterol, Triglycerides and HDL cholesterol concentrations were significantly high in all the groups during summer than in winter and monsoon. No significant difference could be observed in the values of LDL cholesterol. High Sodium values with lower Mg was observed in winter among all the groups of animals. There was no significant difference in Serum Calcium and Phosphorus levels. High AST, ALT and ALP enzyme concentrations during summer with no alterations in GGT was observed in all the groups. SDS PAGE pattern of all the animals during all the seasons showed same pattern with six prominent bands at 22, 27, 40, 53, 66 and 130 kDa. 2D-PAGE analysis of serum protein samples of the Punganur cattle showed More than 415 spots. The present study revealed presence of 12, 23 and 21 differentially expressed protein spots in males; 18, 39 and 37spots in females between summer vs monsoon, summer vs winter and monsoon vs winter respectively.
  • ThesisItemOpen Access
    A STUDY ON SOME OXIDATIVE-STRESS AND ANTIOXIDANT MARKERS IN PUNGANUR CATTLE DURING DIFFERENT SEASONS
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2021-05) SHALOME, SANDRAPATI; ASWANI KUMAR, K (MAJOR); ESWARA PRASAD, P; SRINIVASA PRASAD, CH
    Zebu cattle are reported to be more heat tolerant than crossbred cattle. It is hypothesised that the oxidative status of zebu and crossbred cattle might be compromised differentially to heat stress. Environmental conditions vary significantly during different seasons in tropical climate. The present study was conducted in three different seasons, (summer, Monsoon and winter) in sixty (60) apparently healthy Punganur cattle aged 2 years and above, maintained at Livestock Research Station, SVVU, Palamaner, Chittoor with an objective to assess changes in different markers of oxidative stress, HSP 70 and associated hormones. The mean THI values in the present study were observed to be significantly (p<0.05) higher during summer (79.03±1.22) than monsoon (70.93±1.17). The mean concentrations of glucose values (mg/dl) in males, females and total herd in the present study were significantly low in summer compared to monsoon and winter. Heat stressed cows become hypersensitive to insulin, and will reduce or block adipose mobilization and increasing glucose ‘burning’ in an attempt to minimize metabolic heat production The mean values of TBARS, Catalase (CAT), Superoxide dismutase (SOD) and Glutathione peroxidase (GPx) in males, females and total herd were found to be significantly (P<0.05) higher in summer season compared to monsoon and winter. The activity of Glutathione-S-Transferase in males, females and total herd was found to be insignificant in all the three seasons that were studied. The reduced glutathione values in males, females and total herd were found to be significantly higher in summer (P<0.05) compared to monsoon and winter. The concentration of serum Heat Shock Protein 70 (HSP 70) in males, females and total herd were significantly (p<0.05) higher in summer compared to monsoon and winter, indicating Cellular response of the animals to stressors. The mean concentration values of serum cortisol in males, females and total herd of Punganur cattle were significantly (p<0.05) higher in summer compared to monsoon and winter. Serum triiodothyronine (T3) and Thyroxine (T4) activity in males, females and total herd of Punganur cattle showed a significantly lower (p<0.05) levels in summer compared to monsoon and winter. The trends of T3 and T4 hormones in the present study during different seasons indicates the altered metabolism during different seasons.
  • ThesisItemOpen Access
    CONJUGATION OF ANTIBIOTIC RESIDUES PREVALENT IN MILK WITH CARRIER PROTEIN AND RAISING OF POLYCLONAL ANTIBODIES AGAINST THEM
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2012-08) SAMPATH KUMAR, B; ASHOK, VASILI; ESWAR PRASAD, P; Krishnaiah, N
    ABSTRACT : Antibiotics are the drugs used to kill or inhibit the growth of bacteria. Presence of antibiotic residues in milk has significant public health importance as they have adverse side effects on the health. The present study was undertaken to produce polyclonal antibodies against the most commonly used antibiotics in large animal practice so that a diagnostic test can be developed to screen milk samples for the presence of antibiotic residues. Ampicillin, ceftiofur, enrofloxacin and gentamycin were conjugated with carrier protein BSA by carbodiimide method in which EDC was used as a crosslinking agent. Successful conjugation was confirmed by SDS PAGE in which antibiotic conjugates revealed high molecular weight protein bands of 90kDa, 85kDa, 78kDa, 74kDa for gentamycin, ampicillin, ceftiofur and enrofloxacin respectively in contrast to 68kDa band of BSA. Sprague Dawley rats aged 7-8 weeks were immunized with the antibiotic-BSA conjugates by subcutaneous route according to the standard immunization schedule. A total of four blood samplings were done from each group (3 animals) as follows: first three samples serially at 15 days time interval after 1st immunization plus 1st booster, 2nd booster, 3rd booster and the 4th sampling one and half month after the third booster. The mean total protein content in the antisera of ampicillin, ceftiofur, enrofloxacin and gentamycin groups were 12±1.15 g/dL, 30±1.20 g/dL, 24±1.73 g/dL and 8.25±0.20 g/dL respectively and in the control group serum it was 6.66±0.01 g/dL. The mean albumin concentrations were 3.44±0.020 g/dL, 3.36±0.173 g/dL, 3.21±0.003 g/dL & 2.93±0.014 g/dL for ceftiofur, ampicillin, enrofloxacin and gentamycin groups respectively compared to 3.36±0.173 g/dL in the control group. The mean globulin content in the antisera of ampicillin, ceftiofur, enrofloxacin and gentamycin groups were 8.64±0.10 g/dL, 26.56±1.18 g/dL, 20.79±0.08 g/dL and 5.32±0.05 g/dL respectively and it was 3.30±0.04 g/dL in the control group. A/G ratios were less than the normal value (>1) in antisera of ampicillin, ceftiofur, enrofloxacin and gentamycin groups. Indirect ELISA was standardized with antibiotic-casein conjugate as coating antigen to detect antibody titres. The optimum antigen concentration was chosen as 20ng/ml (which was in the range of Maximum residue limits) and optimum antiserum dilution was chosen as 1/100 by checker board titration. The antibody titres in the test samples of 4 immunization cycles for each antibiotic were determined by indirect ELISA at various serum dilutions ranging from 1/100 to 1/6400. Cut off value of 0.3 was calculated from PNT base line constructed with M+3SD values of control serum samples. All the four antibiotic-BSA conjugates elicited positive immune response in rats. The immune response in BSA-ampicillin conjugate immunized rats was maximum with the highest antibody titre of 2.577 (OD450) followed by BSA-ceftiofur conjugate immunized rats, BSA-enrofloxacin conjugate immunized rats and BSA-gentamycin immunized rats with OD450 values of 2.072, 1.723, and 0.928 respectively at the antiserum dilution of 1/00. These polyclonal antibodies are the efficient tools for developing immunoassays to detect specific antibiotic residues in biological fluids like milk and urine samples
  • ThesisItemOpen Access
    DIFFERENTIAL EXPRESSION OF CHICKEN BRAIN PROTEINS DURING DEVELOPMENT
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2010-06) NAVEEN SWAROOP, M; ASHOK, VASILI(MAJOR); NARSIMHA REDDY, Y; ESWAR PRASAD, P
    ABSTRACT: Understanding the proteins role and their spatiotemporal relationships in the development of brain will enrich the knowledge about structure – function relationships, control of productive traits, cognitive functions, behavior, artificial intelligence and neurological diseases. This study explores the different proteins involved in the development of the brain. Brains from Vanaraja breed embryos were collected from days 2 to 13continuously; from days 15,17,19,21 and from 6weeks birds. Brains were dissected under stereozoom microscope, collected in tris buffer pH 8.3, homogenized and centrifuged in refrigerated centrifuge at 10,000 rpm for 20 min. The supernatants were collected and analyzed by SDS PAGE (5% Stacking gel and 12% resolving gel). The proteins were differentially expressed during different developmental stages. 27 prominent differentially expressed proteins were identified. There was a progressive increase in the expression levels of higher molecular weights with advancing age. During early development of brain, may be low molecular weight proteins are playing a vital role in neural induction, anterio –posterior patterning, dorsovental patterning and cell differentiation. Based on molecular weights, in the light of earlier literature some of these proteins may be kinectin (160kDa), aggrecan (145kDa), PLC-β4 (130kDa), amyloid precursor proteins (120-123kDa), anosmin1 (100kDa), β catenin (92-102kDa), Brevican (97kDa), clusterin (80kDa), hyaluran receptor for motility (75kDa), heat shock protein (73kDa), GPI brevican (66kDa), occludin (60kDa), Glypican (58kDa), stanniocalcin (50kDa), synaptic vesicle protein (34kDa), claudin (22-25kDa), shhN protein (20kDa), FGF (16kDa), myelin basic protein (14kDa). Further research has to be done using gradient PAGE, 2D PAGE, immunohistochemistry and other molecular biological techniques like gene knockout and microarray to confirm the identity and to know the functional role and spatio-temporal relations of these proteins during brain development.
  • ThesisItemOpen Access
    EFFECT OF BISPHENOL-A ON OXIDATIVE STRESS DURING EMBRYONIC DEVELOPMENT IN CHICKS
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2015-01) SRAVANI, J; PADMAJA, K(MAJOR); ESWARA PRASAD, P; PUNYA KUMARI, B
    ABSTRACT: An experiment was conducted to study the effect of Bisphenol-A (BPA) on oxidative stress during embryonic development in chicks. Fertile eggs were procured and eighty (80) eggs in two sets were assigned randomly into 4 groups of20 each. Group I served as control and received 10 μl of distilled water, groups II to IV were treated with BPA dissolved in distilled waterin three concentrations of 100μM, 250μM and 500μM respectively. A single dose of 10 μl volume injected into the aircell using microsyringe separately to 11thday old and 14th day old chick embryos. The developed embryos were sacrificed after 24 and 48 hours of BPA exposure and liver and brain tissues were collected and preserved at -20ºC for estimation of Thiobarbituric acid reacting substances (TBARS), antioxidant enzyme activities (Superoxide dismutase (SOD),Catalase (CAT),Glutathione peroxidise (GPx) and Glutathione-S-transferase (GST)), Reduced glutathione (GSH), Deoxy ribonucleic acid (DNA), Ribonucleic acid (RNA) and protein levels. The liver and brain tissues were preserved in 10% formalin for histopathological studies. The results showed a significant dose dependent increase in TBARS after 24 and 48 hours in both 11th and 14th day treated embryos in liver and brain tissues. Glutathione levels were elevated after 24 and 48 hours in both liver and brain tissues of 11th and 14th day treated embryos. Protein levels were not much altered in both liver and brain tissues, whereas a significant induction in GPx activity was observed after 24 and 48 hours in group IV followed by group III, group II compared to group I in both 11th and 14th day BPA exposed chick embryos. A significant increase in GST activity was observed after 48 hours compared to 24 hours with BPA treatment in both 11th and 14th day old chick embryos. A significant increase in SOD activity was noticed after 24 hours in all the treatment groups compared to control in liver of both 11thand 14th day BPA administered chick embryos. Whereas CAT activity was not altered much after 24 hours, but a significant increase was noticed after 48 hours in liver and brain of 11th and 14th day BPA treated chick embryos. A dose dependent decrease in DNA levels was observed in all treatment groups compared to control after 24 hours in both 11th and 14th day BPA treated chick embryos compared to control. A significant dose dependent decrease in RNA levels was observed after BPA exposure compared to control after 24 hours in both 11th and 14thday BPA treated chick embryos. In histopathological sections, degenerative, necrotic changes with mononuclear cell infiltration, congestion and proliferation of fibroblasts are prominent in liver of both 11th and 14th day BPA treated chick embryos. Neuronophagia, satellitosis, extensive demyelinating changes, proliferation of the capillaries, perineuronal vacuolation was observed in brain of BPA treated chick embryos. The study concludes that the toxic effects of BPA are dose dependent and are inversely effective with age during the embryonic development in chicks.