Attenuation of lentogenic Newcastle disease virus strain B-1 by cold adaptation

Abstract

The Hitchner B-1 strain of Newcastle disease virus was plaque-cloned and then serially passaged 36 times in specific-pathogen-free (SPF) chicken embryos incubated at two different temperatures. Virus passaged at a reduced temperature (29 C) was identified as cold-adapted (Ca) and virus passaged at the normal temperature (37 C) was designated non-cold-adapted (non-Ca). The Ca and non-Ca B-1 viruses were compared with the parent B-1 and a commercial B-1 vaccine. In vitro Ca B-1 characteristics included adaptation for more rapid growth at 29 C and the aquisition of temperature sensitivity indicated by substantially reduced growth at 41 C, properties not seen with non-Ca B-1. Embryo mean death times for the Ca virus (140 hr) were longer than for non-Ca B-1 (107 hr) and parent B-1 (121 hr) viruses. The Ca virus retained a rapid (< 2 hr) hemagglutination (HA) elution rate but lost the property of binding the monoclonal antibody AVS-I typical of other B-1 strains. The pathogenicity of the Ca B-1 strain was compared to the non-Ca B-1, parent B-1 strain, and a commercial B-1 strain vaccine in 1-day-old broiler-type chickens. Pathogenicity was evaluated by assessing the severity of respiratory disease signs and the incidence of airsacculitis, perihepatitis, and pericarditis lesions in inoculated chicks. A respiratory disease index was calculated for each B-1 strain based on daily observation scores that determined the presence or absence of disease signs (coughing, rales, labored breathing, death) from 1 to 14 days following intratracheal inoculation with 10(6) 50% egg infective doses of virus per chick. The lower respiratory disease index obtained for the Ca B-1 strain (0.075) indicated it was less pathogenic than the commercial B-1 vaccine (0.296) and the non-Ca (0.478) and parent (0.521) B-1 strains. Ca B-1-infected chicks had only a 5% incidence of air sac lesions, compared to chicks given non-Ca (65%), Hitchner B-1 (65%), or a commercial B-1 vaccine (30%). Immunogenicity tests performed in 1-week-old SPF leghorn chickens demonstrated that Ca B-1 induced complete protection when administered intraocularly as a single entity. However, when Ca B-1 was given in combination with a modified live infectious bronchitis virus vaccine, chickens were only partially protected (60-75%) against Texas GB strain-induced neurotropic velogenic Newcastle disease.