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Govind Ballabh Pant University of Agriculture and Technology, Pantnagar
After independence, development of the rural sector was considered the primary concern of the Government of India. In 1949, with the appointment of the Radhakrishnan University Education Commission, imparting of agricultural education through the setting up of rural universities became the focal point. Later, in 1954 an Indo-American team led by Dr. K.R. Damle, the Vice-President of ICAR, was constituted that arrived at the idea of establishing a Rural University on the land-grant pattern of USA. As a consequence a contract between the Government of India, the Technical Cooperation Mission and some land-grant universities of USA, was signed to promote agricultural education in the country. The US universities included the universities of Tennessee, the Ohio State University, the Kansas State University, The University of Illinois, the Pennsylvania State University and the University of Missouri. The task of assisting Uttar Pradesh in establishing an agricultural university was assigned to the University of Illinois which signed a contract in 1959 to establish an agricultural University in the State. Dean, H.W. Hannah, of the University of Illinois prepared a blueprint for a Rural University to be set up at the Tarai State Farm in the district Nainital, UP. In the initial stage the University of Illinois also offered the services of its scientists and teachers. Thus, in 1960, the first agricultural university of India, UP Agricultural University, came into being by an Act of legislation, UP Act XI-V of 1958. The Act was later amended under UP Universities Re-enactment and Amendment Act 1972 and the University was rechristened as Govind Ballabh Pant University of Agriculture and Technology keeping in view the contributions of Pt. Govind Ballabh Pant, the then Chief Minister of UP. The University was dedicated to the Nation by the first Prime Minister of India Pt Jawaharlal Nehru on 17 November 1960.
The G.B. Pant University is a symbol of successful partnership between India and the United States. The establishment of this university brought about a revolution in agricultural education, research and extension. It paved the way for setting up of 31 other agricultural universities in the country.
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ThesisItem Open Access Studies on virulence gene profiling, serotyping and epidemiology of shigatoxigenic and enteropathogenic e. coli isolated from captive and free ranging wild mammals and birds(G. B. Pant University of Agriculture and Technology, Pantnagar, 2022-08) Shukla, Namita; Rajesh KumarIndia has great wildlife biodiversity and is home to myriad wildlife fauna. Wildlife plays important role in of the planet ecosystem however, it often represents an important risk in emerging zoonosis, scant information is available about the occurrence of zoonotic pathogens in wildlife worldwide. Shigatoxigenic and Enteropathogenic Escherichia coli are two important classes of zoonotic enteric pathogens. Epidemiology of STEC and EPEC is largely unknown in most of the developing countries including India. Recent, studies in have underlined the importance of wildlife surveillance, as large number of emerging zoonotic pathogens are found to be of wildlife origin. Thus, wild animal should thoroughly be monitored, as they can potentially cause a spillover or spillback to human and other domestic animals. In present study, 770 fecal sample were obtained from 598 wild and 172 domestic animal during January 2021 to March 2022 from zoological gardens situated in three different geographical locations of the India and adjoining village near Accanakmar sanctuary Bilaspur Chhattisgarh respectively. Total 515 (86.12%) isolates were identified as E. coli from wild animals subjected to molecular screening for stx1, stx2, eaeA, and ehlyA genes by multiplex PCR. Total 75 (14.56%) isolates were successfully pathotyped and among these isolates STEC/AE-STEC and EPEC virulence genes were detected in 42 (8.73%) and 33(6.40%)E. coli isolates respectively. All 33EPEC isolates were found to be atypical EPEC carrying only the eaeA gene. All 75 STEC / EPEC isolates were serogrouped in29 different serotype, 3 isolates were untypeable (UT), and 1 isolate was found to be rough strain. Serogroup O157 AE-STEC was detected in two isolates from wild animals, one was isolated from red deer and other from peacock. Present study attempts to investigate transmission of STEC and EPEC among domestic animal, poultry and free- ranging wild animal in shared agroecosystem. One AE-STEC O157 isolates each was isolated from peacock, poultry and goat. Which showed similar virulence profile All 75 isolates(STEC and EPEC) were subjected to invitro antibiotic sensitivity assay against 14 commonly used antibiotics (CLSI 2008), which are showing varying resistance to tetracycline Norfloxacin cefuroxime, Gentamicin, Chloramphenicol, Co-Trimoxazole, Ampicillin and Azithromycin, however, no resistance was observed for cephalothin, ciprofloxacin, ceftriaxone, Ticarcillin+ clavulanic acid and Piperacillin + tazobactam, Amikacin. SETC/EPEC isolates (12) from free-ranging wild animals were susceptible to all 14 of antibiotics tested.AE-STEC, STEC and EPEC isolates differed in their ability to form biofilm according to the temperature, time of incubation and surface used and are classified as no biofilm, weak, moderate, or strong biofilm producers. The biofilm formation was more at 37°C than 20°C during initial 24 hrs. incubation and decreased with increasing incubation time at 37°C, as majority of isolates developed highest biofilm mass at 24 hrs., post inoculation. All the AE-STEC, STEC and EPEC isolates were capable to form biofilm. Our findings emphasize the role of wild animals as reservoir of potentially pathogenic STEC and EPEC.ThesisItem Open Access Prevalence, seroprevalence and postvaccinal antibody response of Peste des Petits ruminants virus in goats(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-11) Netam, Amisha; Tewari, AnujPeste des petits ruminants (PPR) is an economically important transboundary disease of sheep and goats caused by Peste des petits ruminants virus (PPRV). In India, the disease is endemic and therefore, Government of India has initiated PPR control program (PPR-CP) under which kids more than 4 months of age are vaccinated. At the same time it is also important to know the seroprevalence of PPR in unvaccinated goats to know the virus circulation. Therefore, this study was conducted to know the seroprevalence of PPR in unvaccinated goats around the Pantnagar region of Uttarakhand and also to understand the virus distribution in the region. In addition, study also included antibody response and kinetics in Pantja goats vaccinated against PPRV. Total 212 serum samples from goats were collected randomly from various villages from three district (Udham singh Nagar, Nainital, and Almora) of Uttarakhand. Serum samples were tested for anti-PPRV antibody by a commercially available kit from IDvet. 41 animals from various villages were found positive with a prevalence rate of 19.33%. At the same time, PPR outbreaks were also reported from the Pantnagar area. Blood, nasal, oral and rectal swabs were collected from the 19 goats suspected/showing clear sign of PPR. RNA was extracted from the swabs and was subject to one step RT-PCR. The amplified PCR product confirmed PPR in 8 goats with a PPRV prevalence rate 42.10%. Two representative swab samples (one pooled swab and one nasal swab) were subjected to virus isolation in Vero cells. Swabs from both goats showed typical cytopathic effect of PPRV in the first passage and led to complete detachment of the cell monolayer in 48-72 hours in comparison to the vaccine strain Sungri 96 which showed complete cytopathic effect in 4-5 days without complete monolayer detachment. Post vaccination antibody response in Pantja goats vaccinated against PPRV varied from 7-10 days and the antibody response was maintained upto 91 days. Hence the present study signifies that PPRV is circulating in the Tarai region of Uttarakhand and there is an urgent need of mass vaccination to increase the herd immunity to substantial level.ThesisItem Open Access Studies on immunogenicity of recombinant fiber protein of Inclusion Body Hepatitis-Hydropericardium Syndrome (IBH-HPS) virus with special reference to formulation of a candidate subunit vaccine(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-02) Pandey, Garima; Rajesh KumarIn the present study, recombinant fiber protein of strain FAdV- 2/11 of Fowl adenovirus D isolate PANTNAGAR/HA-14/R-21 was studied for its immunogenicity in terms of effect of dose and adjuvant on the immune response of chicken against IBH-HPS virus. SPF chickens were immunized with different doses of recombinant fiber protein with FCA and 25μg/bird dose provided best protection. In second experiment, broilers were immunized with 25μg/bird along with different adjuvants viz ; montanide, resiquimod, saponin and FCA and different immune parameters were studied. Macrophage function test revealed that resiquimod group has released maximum concentration of nitric oxide. Cell mediated and humoral immune responses were analyzed by cutaneous basophil hypersentivity test, lymphocytes proliferation test and serum neutralization test, respectively. Maximum thickness of foot web was observed in montanide group at 72hrs post inoculation of DNCB. Montanide group showed maximum Tcell response at 21st DPI. Neutralization index of montanide group was highest at 28th DPI. Viral DNA in faeces was detected in all groups at 7th DPC, in montanide group 3 out of 6 faecal samples were positive while on 10th DPC only challenged control group was positive for viral DNA in faeces. 25μg/bird dose with montanide showed best immune response in broilers against challenged with virulent FAdV-2/11. Immunoinformatics analysis of Fiber protein of FAdV-2/11 revealed 21 continuous B cell epitopes with 13 epitopes having surface accessibility and 19 epitopes were antigenic as predicted by BepiPred, Emini surface accessible and Kolaskar and Taogankar antigenicity method, respectively. Out of four models predicted by SWISS MODEL, model 1 was best and verified by different servers. Three discontinuous epitopes were predicted by Ellipro tool. Sixteen epitopes, strongly linked with the MHC alleles were determined by MHC Class-I binding tool and six core peptides have been predicted by MHC Class-II binding tool. Secondary and tertiary structures were predicted by PesiPred V4.0 and Phyre2 tool, respectively. Physico-chemical properties of fiber protein were predicted by ProtParam tool. 0.5002 antigenicity score was predicted by VaxiJen v2 server and thus, fiber protein was proved antigenic and immunogenic. Therefore, fiber protein can be used for development of promising peptide vaccines. Present study concludes that a recombinant subunit vaccine candidate containing recombinant fiber protein (25μg/bird) with montanide as adjuvant may be formulated for further field trials.ThesisItem Open Access Egg derived antibodies (IgY) against Outer membrane proteins of multi-drug resistant Salmonella Typhimurium: Production and evaluation of therapeutic potential(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-03) Tiwari, Aakanksha; Rajesh KumarKeeping in view, the emerging problem of anti microbial resistance in Salmonella Typhimurium, the present research work was carried out to produce and assess a therapeutic alternative against the bacteria. In the present study, attempts were made to revive glycerol stocks of 150 field isolates and 14 isolates were successfully revived. These isolates were characterized culturally and subjected to ABST. The isolate showing resistance against six groups of antibiotics was selected for further study. This isolate was also characterized molecularly and serotyped as Salmonella Typhimurium with antigen types- 4,5,12:i:1,2. After confirmation of the culture, OMPs were isolated with a concentration of 7.28 mg/ml and SDS-PAGE analysis revealed prominent OMP bands ranging from 11-90 kDa. OMPs were used for hyperimmunizing the RIR layers with a suitable adjuvant subcutaneously. Eggs were collected and IgY was isolated by dextran sulphate method. The purified IgY preparation revealed bands of 63 kDa and 27 kDa in SDS-PAGE, corresponding to the heavy and the light chains. Total protein concentration of IgY preparation by Lowry method was 14.246 mg/ml and specific IgY concentration by RID was 12.023 mg/ml, indicating 84.40% purity. The specificity of IgY against the OMPs was indicated by positive reaction in AGID, CIE, Western blot and Dot Enzyme Immunoassay. Titre of antibody in serum and egg yolk at weekly basis was also determined by ELISA. The maximum antibody titre was observed at 13th week and 11th week in the egg yolk and serum, respectively. It was also observed that IgY was stable at temperatures ranging from -40 ̊ C to 37 ̊ C as indicated by ELISA and SDS-PAGE. In-vitro efficacy testing of different concentrations of IgY against the bacteria showed results in a dose-dependent manner and significant difference was observed between the different combinations. Massive adherence and penetration of bacteria in Vero cells was observed in the negative and culture control unlike in the IgY treated bacterial Vero cells. In vivo experiment in mice, prophylactic group showed no mortality and bacterial count in the faecal swabs and in the organs was significantly less. It indicates that prophylactic activity of IgY is much stronger than the therapeutic activity.ThesisItem Open Access Studies on immunogenicity of recombinant fiber protein of Inclusion Body Hepatitis- Hydropericardium Syndrome (IBH-HPS) virus with special reference to formulation of a candidate subunit vaccine(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-02) Pandey, Garima; Rajesh KumarIn the present study, recombinant fiber protein of strain FAdV- 2/11 of Fowl adenovirus D isolate PANTNAGAR/HA-14/R-21 was studied for its immunogenicity in terms of effect of dose and adjuvant on the immune response of chicken against IBH-HPS virus. SPF chickens were immunized with different doses of recombinant fiber protein with FCA and 25μg/bird dose provided best protection. In second experiment, broilers were immunized with 25μg/bird along with different adjuvants viz ; montanide, resiquimod, saponin and FCA and different immune parameters were studied. Macrophage function test revealed that resiquimod group has released maximum concentration of nitric oxide. Cell mediated and humoral immune responses were analyzed by cutaneous basophil hypersentivity test, lymphocytes proliferation test and serum neutralization test, respectively. Maximum thickness of foot web was observed in montanide group at 72hrs post inoculation of DNCB. Montanide group showed maximum Tcell response at 21st DPI. Neutralization index of montanide group was highest at 28th DPI. Viral DNA in faeces was detected in all groups at 7th DPC, in montanide group 3 out of 6 faecal samples were positive while on 10th DPC only challenged control group was positive for viral DNA in faeces. 25μg/bird dose with montanide showed best immune response in broilers against challenged with virulent FAdV-2/11. Immunoinformatics analysis of Fiber protein of FAdV-2/11 revealed 21 continuous B cell epitopes with 13 epitopes having surface accessibility and 19 epitopes were antigenic as predicted by BepiPred, Emini surface accessible and Kolaskar and Taogankar antigenicity method, respectively. Out of four models predicted by SWISS MODEL, model 1 was best and verified by different servers. Three discontinuous epitopes were predicted by Ellipro tool. Sixteen epitopes, strongly linked with the MHC alleles were determined by MHC Class-I binding tool and six core peptides have been predicted by MHC Class-II binding tool. Secondary and tertiary structures were predicted by PesiPred V4.0 and Phyre2 tool, respectively. Physico-chemical properties of fiber protein were predicted by ProtParam tool. 0.5002 antigenicity score was predicted by VaxiJen v2 server and thus, fiber protein was proved antigenic and immunogenic. Therefore, fiber protein can be used for development of promising peptide vaccines. Present study concludes that a recombinant subunit vaccine candidate containing recombinant fiber protein (25μg/bird) with montanide as adjuvant may be formulated for further field trials.ThesisItem Open Access Studies on immune status of indigenous and exotic breeds of domestic fowl(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Yadav, Renuka; Rajesh KumarPresent study was carried out for comparative evaluation of some indigenous and exotic breeds of domestic chicken with regard to the innate immune response, adaptive immune response and resistence to infection following challenge with infectious agent. One exotic (RIR) and two indigenous breeds (Kadaknath, Uttara fowl) were used for study. All the birds were reared in deep litter system with ad-libtum feed and water throughout the experiment. The experiment lasted for 60 days. Significant difference was observed among parameters studied viz; mean total leukocyte count, differential leukocyte count, leukocrit, serum globulin, A/G ratio, phagocytic activity and opsonocytophagic index, among various breeds of chicken at all stages and all the innate and adaptive immune parameter was significantly higher in RIR compared to Kadaknath and Uttara fowl breed of chicken. The RIR showed highest hemagglutination inhibition titre following NDV immunization till the end of the experimental period followed by Kadaknath and Uttara fowl. Cell mediated immune (CMI) response was also observed to be higher for RIR breed compared to Kadaknath and Uttara fowl. The lymphocyte stimulation index was highest in RIR and the lowest in Uttara fowl. Maximum mortality was observed in Uttara fowl up to 15 days post challenge with a virulent virus and antibody titre was maximum in RIR breed at 3 and 7 days post challenge but at 14 days post challenge kadaknath showed highest antibody titre.ThesisItem Open Access Studies on epidemiology, isolation and molecular analysis of fowl adenoviruses from the domestic chicken with respiratory disease conditions(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Bisht, Ritika; Rajesh KumarFowl adenoviruses (FAdV) are the ubiquitous and cause extensive damage to the poultry industry. They are known to be associated with various disease conditions such as Inclusion body hepatitis, Hydropericardium Syndrome, respiratory disease, tenosynovitis, impaired growth, reduced egg production, aplastic anemia, atrophy of bursa and thymus, enteritis and conjunctivitis in chickens and other birds. Present study was undertaken to study epidemiology, to isolate and perform molecular analysis of fowl adenoviruses from the chickens with the respiratory disease conditions. Natural outbreaks investigated during the study revealed 2 –15% mortality among the affected birds aged 3-6 weeks. General postmortem findings were enlarged and friable liver with necrotic foci, enlargement of kidney, congestion and haemorrhagic trachea, congested lungs. Four isolates of fowl adenoviruses were propagated in primary CEL cell culture. The cytopathic effects were characterized by rounding of the cells, degeneration, increase in the refractive index of the cells and microplaques formation. AGPT, CIEP, IFAT, Dot-ELISA and Sandwich ELISA determined the presence of the virus in infected tissues and cell culture. Intensely basophilic intra nuclear inclusion bodies were also visualized following MGG staining of CEL cells. The genomic DNAs were extracted from the infected CEL cells and PCR was carried out for amplification of L1 region of the hexon gene showing amplicons of the ~900 bp. On digestion with MluI, three isolates were digested and the two bands of 400 and 500 bp were visualized and with the enzyme StyI, only one isolate was digested and two bands of 480 and 420 bp were obtained. None of the isolates were digested by the enzymes BglI, BsiWiI and ScaI. RE analysis revealed that isolates belong to two serotypes 5 (R-66) and 8 (R-53, R-43, R-63). Thus, it can be concluded that these serotypes are involved in respiratory disease of domestic chickens in India.