Vaccination is a good strategy for the prevention of avian influenza virus. In this research Gamma Irradiated Avian Influenza (Sub type H9N2) Vaccine (GAIV) was prepared by 30 kGy irradiation and used for vaccination of broiler chickens. The purpose was a comparison of immune responses in the two routes of administration for the GAIV vaccine; intranasal and subcutaneously, use of Montanide ISA70 and Trehalose accompanied with irradiated vaccine and compare with formalin vaccine. The Influenza Virus A/Chicken/IRN/Ghazvin/2001/H9N2 was irradiated and used for vaccine formulation, and formalin inactivated AIV was used as conventional vaccine. Chickens were vaccinated by GAIV with and without Trehalose, GAIV and formalin vaccines with ISA70, two routes of administration were intranasal and subcutaneously. All the vaccinated chickens showed a significant increase in antibody titration. The most significant increase of antibody titration was in irradiated vaccine plus Trehalose groups intranasal and subcutaneously. After the first and second intranasal vaccination, the amount of IFN-gamma increased in the irradiated vaccine plus Trehalose group compared to other groups. However, most of the vaccinated groups did not show any significant increase of IFN-α concentration. Histopathological examination revealed lymphocyte infiltration (++), foci dispersed of hemorrhage and edema in intranasal vaccination groups and in addition to these, thickening of alveolar septa was observed in the injection groups. GAIV vaccine can be a good candidate for vaccine preparation, and Trehalose as a stabilizer protects viral antigenic proteins, also makes more absorbance of antigen by the inhalation route. In vaccinated chickens the ulcers in injected vaccines were lower than intranasal vaccines. HIGHLIGHTS  Gamma Irradiated Avian Influenza (H9N2) Vaccine was prepared by 30 kGy irradiation.  Broiler chickens were vaccinated by two routs; intranasal and subcutaneously.  All the vaccinated chickens showed a significant increase in antibody titration.  The most increasing of immune responses was in irradiated vaccine plus Trehalose. 2 Motamedi-Sedeh, F.; et al.

Avian influenza A H9N2 viruses are circulating in domestic poultry worldwide. In this research a low-pathogenic AIV H9N2 was multiplied on MDCK cell line and SPF eggs and irradiated by a Nordian gamma cell instrument. Irradiated and non-irradiated AIV samples were titrated by TCID50 and EID50 methods, respectively. Haemagglutinin antigen was analysed by Haemagglutinin test. Infectivi­ty of irradiated virus samples was determined by cell culture and egg inoculation methods. The virus titration decreased as the dose of gamma radiation increased. AIV proliferation on cell culture can be inactivated by gamma irradiation at a lower dose of gamma-ray (20 kGy) than the virus inactivation on embryonated eggs (30 kGy). The safety test showed complete inactivation of AIV on allantoic fluid with gamma-ray doses: 30 kGy and 20 kGy for virus on MDCK cells after four blind cultures.