The genetic evolution of highly pathogenic avian influenza (HPAI) in Egypt has developed a new clade H5N1 (2.2.1.2) since 2014. Meanwhile, the new avian influenza virus (AIV) clade mutually with the velogenic Newcastle disease virus (NDV) isolate of genotype VII in Egypt (genotype VII) has resulted in severe economic losses in the broiler industry. An inactivated bivalent vaccine containing H5 (belonging to H5N1 clade 2.3.2) recombinant baculovirus expressed by insect cell (recH5) and egg-based NDV LaSota strain (recH5/NDV vaccine) was evaluated for protection against the challenge of dual HPAIV H5N1 clade 2.2.1.2 and vNDV infection in commercial broiler chickens. Vaccination was performed when chickens were 10 days old, and then birds of the respective groups were challenged with 10 50% egg infective dose per chicken of each virus in 100 µl of allantoic fluid via the intranasal route at 21 days postvaccination in a single or sequential infection of both viruses. Results showed that the recH5/NDV vaccine was able to protect chickens against single or dual challenges of both viruses ranging up to 90%-100%. Unvaccinated chickens have demonstrated 100% mortalities to a single virus challenge. Vaccinated chickens showed significant decreases in both viruses, shedding titers up to <2 log after challenge in comparison with unvaccinated ones. Cessation of viral shedding was obtained at 7 to 10 days postchallenge. The vaccinated chickens showed high hemagglutination inhibition antibody titers >6 log against both H5N1 and NDV antigens at 2 wk postvaccination. The single vaccination of bivalent inactivated recH5-NDV vaccine at 10 days old in commercial chickens has provided significant clinical protective immunity against single or dual challenge with HPAI-H5N1clade 2.2.1.2 and vNDV-genotype VII.

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