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The protective dose of a live recombinant LaSota Newcastle disease virus (NDV)-avian influenza H5 vaccine (rNDV-LS/AI-H5) was determined in broiler chickens with high levels of maternal antibodies against NDV and avian influenza virus (AIV). At hatch the geometric mean titers (GMT) of the chickens' maternal antibodies were 2(5.1) and 2(10.3) for NDV and AIV, respectively. At the time of vaccination the GMT was 2(3.1) for NDV and 2(7.9) for AIV. The chickens were vaccinated with one drop (0.03 ml) in the eye at 10 days of age as is typical under field conditions. The test chickens received 10(4.8), 10(5.8), 10(6.8), or 10(7.8) mean chicken embryo infective doses (CEID50) of the rNDV-LS/AI-H5 vaccine. Control chickens were either nonvaccinated, or vaccinated with 10(5.8) or 10(6.8) CEID50 of a commercial live LaSota NDV vaccine. Birds were challenged with either the Mexican highly pathogenic avian influenza virus (HPAIV) strain A/Chicken/Queretaro/14588-19/95 (H5N2) or a Mexican velogenic viscerotropic (VV) NDV strain. One hundred percent of the chickens vaccinated with the rNDV-LS/AI-H5 vaccine were protected against HPAIV and VVNDV when a challenge dose of 10(6.8) EID50 or higher was administered by eye drop. Birds vaccinated with the LaSota NDV vaccine were protected against VVNDV, but not against HPAIV.

Specific-pathogen-free chickens immunized at 14 days of age with either an inactivated recombinant Newcastle disease virus-LaSota/avian influenza H5 (K-rNDV-LS/AI-H5) vaccine or a killed Newcastle disease/avian influenza whole-virus vaccine (K-ND/AI) were protected from disease when challenged with either A/chicken/Queretaro/14588-19/95 (H5N2), a high pathogenicity avian influenza virus (HPAIV) strain isolated in Mexico in 1995, or with a Mexican velogenic viscerotropic Newcastle disease virus (VVNDV) strain 21 days postvaccination. All nonvaccinated chickens challenged with HPAIV or VVNDV succumbed to disease, while those vaccinated with K-rNDV-LS/AI-H5 or K-ND/AI were protected from severe clinical signs and death. Both vaccines induced hemagglutination-inhibition (HI) antibody responses against NDV and AIV. Antibodies against AIV nucleoprotein were not detected by enzyme-linked immunosorbent assay (ELISA) in birds vaccinated with the inactivated rNDV-LS/AI-H5 vaccine. These chickens became positive for AIV antibodies by ELISA only after challenge with HPAIV. The data clearly indicate that the inactivated rNDV-LS/AI-H5 vaccine confers protection comparable to that of the conventional killed whole-virus vaccine against both NDV and AIV, while still allowing differentiation of infected from vaccinated animals by HI and ELISA tests.