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Thesis

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2017 - 20183
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Subject: Animal Biochemistry × Start date: 2017 ×
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Now showing 1 - 3 of 3
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    ThesisItemOpen Access
    GENOTYPING AND PROTEIN PROFILING OF MILK β-CASEIN VARIANTS (A1 and A2) IN INDIAN BREEDS OF MILCH CATTLE
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2018-03) SRINIVAS, B; ASWANI KUMAR, K(MAJOR); PADMAJA, K; PUNYA KUMARI, B
    ABSTRACT: The exotic and cross bred cattle milk contain A1 β-casein variant, whereas indigenous cattle milk contains A2 β-casein variant. They have structural diversity at 67th amino acid residue proline in A2 which is replaced by histidine in A1 during course of evolution. The histidine at 67th amino acid residue generates a proteolytic cleavage site in A1 leading to the production of betacasomorphin-7 (BCM-7) that is known to be a risk factor for several human diseases. The serious health concern associated with A1 milk consumption endures an urgent need to select cattle breeds with higher genotypic and allelic frequencies of A2 and conserve indigenous cattle breeds through breeding programmes. In this context, the present study was undertaken to screen seventy animals belonging to five indigenous cattle breeds of India viz., Deoni, Sahiwal, Ongole, Punganur, Malnad Gidda and Holstein Friesan cross (twelve from each breed and ten from Deoni) to characterize β-casein variants (A1 and A2) geotypically by PCR-RFLP with TaqI restriction enzyme and also to study milk β-casein variants (A1 and A2) by IEF. PCR-RFLP with Taq1 restriction enzyme yielded two genotypes as A1A2 (251 and 213 bp) and A2A2 (251 bp) in all indigenous cattle breeds studied, whereas only heterozygous (A1A2) genotype was found in the HF crossbreds. The genotypic frequency of A1A2 heterozygous variant of exon 7 of CSN2 gene was the highest for HF cross (1.0) followed by Sahiwal (0.75), Deoni (0.58), Malnad Gidda (0.40), Punganur (0.17) and Ongole (0.083). Whereas, for A2A2 homozygous variant was the highest for Ongole (0.92) followed by Punganur (0.83), Malnad Gidda (0.60), Deoni (0.42) and Sahiwal (0.25). The A1 allelic frequency of exon7 of CSN2 gene was the highest in HF cross (0.50) followed by Sahiwal (0.38), Deoni (0.29), Malnad Gidda (0.20), Punganur (0.083) and Ongole (0.042), whereas the highest A2 allelic frequency was observed in Ongole (0.96) followed by Punganur (0.92), Malnad Gidda (0.80), Deoni (0.71), Sahiwal (0.63) and HF cross (0.50). The IEF gel electrophoresis pattern of milk β-casein protein showed two distinct bands corresponding to A1 and A2 β-casein protein variants at pI values 5.22 and 5.14 respectively and this polymorphism correlated positively with the results of PCR-RFLP with TaqI restriction enzyme. The present study unveils the existing fact that there is incorporation of exotic germplasm in native cattle breeds to varying degrees in different breeds and emphasizes an immediate need for genotypic screening of bulls for β-casein gene variant A2 for documentation and further measures to improve the indigenous breeds with appropriate breeding strategies.
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    ThesisItemOpen Access
    A COMPARATIVE STUDY ON HAEMATOLOGICAL AND BIOCHEMICAL PARAMETERS IN NORMAL CYCLIC, REPEAT BREEDING AND ANOESTRUM CROSSBRED MURRAH BUFFALOES
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2018-02) ANITA, K; PADMAJA, K(MAJOR); PRASADA RAO, T; PUNYA KUMARI, B
    ABSTRACT: The present study was planned to assess the levels of haematological parameters and biochemical constituents in normal cyclic, repeat breeding and anoestrum crossbred Murrah buffaloes in drought area Proddatur in YSR Kadapa district of Andhra Pradesh. For this study a total of 60 blood samples were collected from each 20 healthy, 20 repeat breeding and 20 anoestrum buffaloes brought to the Department of Veterinary Clinical Complex, College of Veterinary Science, Proddatur and from various Veterinary dispensaries and Veterinary camps conducted in and around Proddatur. The variations in circulating levels of biochemical constituents and haematological parameters have been recorded in normal cyclic, repeat breeding and anoestrum crossbred Murrah buffaloes. The results showed significant decrease in glucose, cholesterol, total protein, albumin, globulin levels and significant increase in triglycerides and blood urea nitrogen in repeat breeding and anoestrum compared to normal cyclic crossbred Murrah buffaloes. Alkaline phosphatase and aspartate aminotransferase activities significantly elevated in repeat breeding and decreased in anoestrum compared to normal cyclic crossbred Murrah buffaloes. Whereas, alanine aminotransferase activity significantly decreased both in repeat breeding and anoestrum compared to normal cyclic crossbred Murrah buffaloes. Significant reduction was observed in calcium and phosphorous levels in repeat breeding and anoestrum compared to normal cyclic crossbred Murrah buffaloes. Alterations observed in sodium, potassium and chloride levels among the groups were marginal. The haematological parameters viz., haemoglobin, total erythrocyte count, total leucocyte count, packed cell volume, platelet count, monocytes, eosinophils and neutrophils were significantly decreased in repeat breeding and anoestrum conditions compared to normal cyclic animals. Whereas, lymphocytes and basophils were significantly increased in repeat breeding and anoestrum conditions compared to normal cyclic animals. In repeat breeding and anoestrum conditions increased malondialdehyde and decreased superoxide dismutase and catalase activities noticed compared to normal cyclic crossbred Murrah buffaloes.
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    ThesisItemOpen Access
    ISOLATION AND PURIFICATION OF GLUTATHIONE-S-TRANSFERASE (GST) AND ITS ROLE ON OXIDATIVE STRESS IN HEPATIC TISSUES OF MAREK’S DISEASE AFFECTED CHICKEN
    (SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-01) RAJIV KISHORE, K.V.; PADMAJA, K(MAJOR); ESWARA PRASAD, P; PUNYA KUMARI, B
    ABSTRACT: A study was conducted to isolate and purify Glutathione-S-transferase (GST) and to determine its role on oxidative stress in liver tissues of Marek’s disease (MD) affected chicken. Ten healthy and MD suspected layer birds of 35-40 weeks age for each group were procured from poultry farms in Chittoor district of A.P. The postmortem examination of MD affected birds revealed enlarged liver with discrete grayish nodules of various sizes. Histopathological examination of liver revealed extensive proliferation and infiltration of pleomorphic cells. The purified DNA samples from liver tissue of MD suspected birds were further used for PCR analysis which was found to be positive yielding a 314 bp product indicating the presence of two copies of 132 bp tandem repeat and a 50 bp primer sequence. The liver samples positive for MD were used for further analysis. The results showed a significant increase of TBARS (179.82%) in liver of MD affected chicken compared to control. Glutathione (GSH) levels were significantly reduced (52%) in liver of MD affected chicken compared to control. A significant decrease in SOD (32%), catalase (43%), GPx (55%) and GST (51%) activities were observed in MD affected chicken compared to control. The GST enzyme was isolated, purified and characterized from liver tissue of both healthy and MD affected layer chicken using glutathione agarose affinity chromatography. The enzyme was purified to 2.38 fold with a yield of 32.27% in healthy chicken whereas in MD affected chicken it was purified to 4.54 fold with a yield of 41.98%. The analysis of active fractions using SDS-PAGE showed GST-α expression with a molecular weight of 25.6kDa in both healthy and MD affected liver. In addition GST-π was expressed in MD affected chicken with a MW of 27 kDa. These results confirm that Marek’s disease in chicken induces oxidative stress which in turn leads to alterations in antioxidant enzymes. The oxidative stress induced in liver is responsible for GST π expression as a defense mechanism in MD affected chicken.