Rajesh KumarPandey, Garima2021-09-142021-09-142021-02https://krishikosh.egranth.ac.in/handle/1/5810175710In 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.EnglishThesis