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Birsa Agricultural University, Ranchi

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2013 - 20194
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Subject: Veterinary Microbiology × End date: 2019 × Start date: 2013 × Type: Thesis ×
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    ThesisItemOpen Access
    Comparative Study on Isolation and Identification of Methicillin Resistant and Non Methicillin resistant Staphylococcus Aureus from Mastitic Milk
    (Birsa Agricultural University, Ranchi, Jharkhand-6, 2019) Kumari, Sweeta; Prasad, Arun
    Comparative Study on Isolation and Identification of Methicillin Resistant and Non Methicillin resistant Staphylococcus Aureus from Mastitic Milk
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    ThesisItemOpen Access
    STUDIES ON IMMUNOMODULATOY EFFECT OF ETHANOLIC EXTRACT OF Trigonellafoenum-graecumLEAVES IN BROILER CHICKS
    (Birsa Agricultural University, Ranchi, Jharkhand-6, 2018) MURMU, SUNITA KUMARI; Prasad, Arun
    On the basis of above finding it can be concluded that the dose 50mg/kg b.wt and 100mg/kg b.wt of ethanolic extract of fenugreek leaves has immunomodulatory effect in broiler chicks, but as the dose increased above from the range of 200 to 400 mg/kg b.wt the toxicity has been observed in liver and kidney which is evident by histopathlogy. Effect of ethanolic extract of fenugreek leaves of treatment on blood biochemical profile showed a mild increase in total serum protein and total serum globulin indicating better humoral immune response. The effect of ethanolic extract of fenugreek leaves showed mild increase in hemoglobin, PCV, TEC. Maternal haemagglutination inhibition titre against NDV vaccine was detectable only up to 21 days of age in non-vaccinated control group of chicks. Food Conversion Ratio(FCR) of higher weight gain per bird at the sale counter fetching more money to farmers.
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    ThesisItemOpen Access
    In Vitro Expression of Recombinaat Buffalo Lactoferrin Gene and its Antibacterial Activity
    (Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2013) Kumari, Namita; Prasad, Arun
    Lactoferrin (Lf) is an iron binding glycoprotein having many biological activities, such as facilitating iron absorption and having antimicrobial and anti-inflammatory effects. Therefore, the present study was undertaken to characterize the buffalo Lf at the genic level and express N-terminal of buffalo Lf in E. coli system to see its antibacterial activity. For that, total RNA was isolated from the mammary gland of lactating buffalo and reverse transcription-polymerase chain reaction (RT-PCR) was carried out for first strand cDNA synthesis. Buffalo lactoferrin gene of 2.2 kb was amplified using gene specific primers and cloned into pTZ57R/T cloning vector using DH5α strain of E. coli. The positive-negative selection of colony was done based on differential colour development by LacZ selection. Positive clones of buffalo Lf gene cDNA were sequenced and were analyzed with the help of laser gene software and submitted to NCBI GenBank (Acc. No. JF825526). Sequencing of the selected representative clones revealed that nucleotide sequence of buffalo Lf contained an ORF of 2127 bp encoding a precursor peptide of 708 amino acids with a molecular weight of 77.65 kDa and isoelectric point of 7.979. Comparative study of buffalo Lf with other reported livestock available in GenBank showed that buffalo Lf had higher homology both at nucleotide and amino acid level with cattle. The phylogenetic analysis also indicated that buffalo Lf had a close relationship with that of cattle. To see the antibacterial activity of in vitro expressed buffalo Lf, 150 bp of Nterminal of buffalo Lf was amplified and cloned in pTZ57R/T cloning vector. Then it was further sub-cloned into the expression vector pQE30 and expressed in E. coli strain (SG13009). After induction with IPTG, the target fusion protein was successfully expressed and identified by SDS-PAGE and Western blotting. The protein purified using His-tag had a molecular weight of 7.0 kDa. Further the expressed buffalo N-Lf protein displayed bactericidal activity after activation by enzymatic proteolysis using porcine pepsin against S. aureus and E. coli comparable to commercially available bovine Lf. The successful expression of functionally active and intact buffalo N-Lf in this study can be used to study biochemical antimicrobial interaction and has the potential to be used as an immunomodulator in future, particularly against infectious diseases like mastitis, thus having significant impact on buffalo productivity.
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    ThesisItemOpen Access
    DEVELOPMENT AND STANDARDIZATION OF DOT-ELISA FOR RAPID AND RELIABLE DIAGNOSIS OF NEWCASTLE DISEASE, INFECTIOUS BRONCHITIS AND INFECTIOUS BURSAL DISEASE IN POULTRY
    (Birsa Agricultural University, Kanke, Ranchi, Jharkhand, 2016) Kumari, Anuradha; Prasad, Arun
    The present work was taken upon with an objective to develop a cheap, sensitive and ready to use technique even in field against ND, IBD and IB. For this development several stages of dot-ELISA has been standardized and the developed dot-ELISA may be helpful for recording the seroprevalence of ND, IBD and IB infections. For dot-ELISA under field conditions following quantification were found to be effective in achieving best results, (A) Antigen standardization ND antigen: 11.25μg Lasota/F1 strain of virus in 0.9 μl of diluent, IBD antigen: 15μg BursaB2K/Gumboro strain of virus in 1.2 μl of diluent, IB antigen: 6.25μg Georgia strain of virus in 0.5 μl of diluents. (B) Blocking solution standardization ND: skimmed milk (1%) + gelatin (0.5%) +BSA (0.5%) IBD: skimmed milk (1.5%) + gelatin (0.5%) +BSA (1%) IB: skimmed milk (2%) + gelatin (1%) (C) Sera standardization ND: 1:40 IBD: 1:10 IB: 1:20 (D) Conjugate standardization ND: 1:800 IBD: 1:500 IB: 1:1100 Standard dot-ELISA developed for ND, IBD and IB was compared with ELISA and AGID. For ND and IB, The result of dot-ELISA when compared with the results of ELISA has slightly lower value whereas dot-ELISA when compared with AGID has showed higher value. Sensitivity and specificity are very important parameter to judge accuracy of the test. The sensitivity and specificity of dot-ELISA calculated in present study was compared with result of ELISA was 68.97% and 61.76% and with AGID which was 95.74% and 82.22% respectively for ND. Similarly for IBD, the sensitivity and specificity of dot-ELISA calculated in the scenario was compared with result of ELISA as 67.57% and 81.82% and with AGID as 88.89% and 94.64% respectively. The sensitivity and specificity of dot- ELISA for IB was compared with result of ELISA as 67.44% and 66.67% and with AGID 93.88% and 67.44% respectively for IB. As far as literature is concerned, ELISA based seoprevalence of ND, IBD and IB has not been reported from Ranchi or from the Jharkhand, therefore it seems to be the first report from this area. In the present study, the overall seroprevalence of ND, IBD and IB on the basis of ELISA, AGID and dot-ELISA was 63.04%, 51.09% and 57.61%; 40.22%, 39.13% and 38.04%; 93.48%, 53.26% and 65.22% respectively. Seroprevalence is very important aspect to determine the threat intensity. Until now we don’t have any economical kit at farm level which helps in regular monitoring. ELISA kit in market as confirmatory diagnosis is quite expensive and it had to be incorporated. Another drawback is that a well equipped lab with trained personnel is required to carry out experiment. Due to these impairments diagnosis at farm level becomes difficult. Diagnosis by dot-ELISA may overcome these short comings and detect the disease at much cheaper rate with appreciable sensitivity and specificity. In comparison to plate ELISA, the developed dot-ELISA can be performed at farm level without sophisticated equipment like ELISA reader, expensive reagents and other specialized instruments. It will prove to be highly cost effective and also suitable for small sample size. The farm personnel can perform the test as per the protocol and read the results easily. Moreover, antibodies to three viruses can be screened at a time which further reduces the cost testing. The present study clearly indicates prevalence of ND, IBD and IB as high, moderate and extremely high in Ranchi respectively. Mere presence of prevailing antibody without the gross pathological lesion indicates mild or subclinical form of viral infection.