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Newcastle disease (ND) is a highly fatal, infectious, viral disease, and despite immunization with live and inactivated vaccines, the disease is still endemic, causing heavy morbidity and mortality leading to huge economic losses to the poultry industry in Pakistan. Therefore, the present study was aimed for the first time in the country at using novel virosomal technology to develop the ND vaccine using an indigenous highly virulent strain of the virus. ND virosome was prepared using Triton X-100, and SM2 Bio-Beads were used to remove the detergent and reconstitute the viral membrane into virosome. Confirmation was done by transmission electron microscopy and protein analysis by SDS-PAGE.  In vitro  cell adhesion property was observed by incorporating green fluorescent protein (GFP), producing plasmid into virosome and  in vitro  cell culture assay. Sterility, safety, and stability of the vaccine were tested before  in vivo  evaluation of immunogenicity and challenge protection study in commercial broiler. The virosome vaccine was administered (30  μ g/bird) at days 7 and 14 through the intranasal route in comparison with commercially available live and inactivated ND vaccines. Results revealed significantly high ( p  < 0.05) and clinically protective hemagglutination inhibition (HI) antibody titers at 7, 14, 21, and 28 days postimmunization with the virosome vaccine in comparison to the negative control. The GMTs were comparable to live and inactivated vaccines with nonsignificant ( p  > 0.05) differences throughout the experiment. Antibody levels increased in all vaccinated groups gradually from the 7 th  day and were maximum at 28 th -day postvaccination. In the virosome-administered group, GMT was 83.18 and 77.62 at 21 st  and 28 th -days postvaccination, respectively. Challenge revealed 100%, 90%, and 80% protection in virosome, live, and inactivated vaccinated groups, respectively. Under given experimental conditions, we can conclude that ND virosome vaccine prepared from the indigenous virus was found to be safe and immunogenic.

Development, Biological Characterization, and Immunological Evaluation of Virosome Vaccine against Newcastle Disease in Pakistan