Evaluation of combined inactivated vaccines against avian influenza subtypes [H5N1, H9N2] and avian infectious Bronchitis in chicken using montanide and paraffin oil adjuvants

Abstract

The goal of the presented work was to prepare and evaluate physical characters andimmunogenicity of a combined inactivated Avian Influenza Virus (AIV) H5N1, H9N2 subtypes andInfectious Bronchitis Virus (IBV), strain D-88 oil emulsion vaccines. The prepared vaccines weresterile and safe. Also, they were ensured to be water in oil (W/O) emulsions using drop test,conductivity (zero mS/cm). Particle size were 950 nm and 1050 nm and dynamic viscosities were19.52 Mpa.s and 39.65 Mpa.s for the vaccine with Montanide™ ISA 71 RVG adjuvant and thevaccine with paraffin oil adjuvant, respectively. They showed stability for 24 months at 4 °C withno separation. Combined inactivated oil emulsion vaccines induced cellular and humoral immuneresponses in vaccinated chicks. The vaccine with Montanide™ ISA 71-RVG adjuvant provided100% protection percent for AIV H5N1 without shedding of AIV H9N2 and IBV in comparison tothe combined vaccine with paraffin oil adjuvant that gave 84% protection percent for AIV H5N1and shedding of AIV H9N2 and IBV occurred 6th day post challenge. Montanide™ 71-RVGadjuvant has the flexible ratio of oil and antigenic media in the vaccine (60:40) allowed using largeamount of virus that showed good impact on its immunogenicity, over the paraffin oil adjuvantwhich has restricted ratio of oil and antigenic media of the vaccine (73:27).