Cold adapted avian pneumovirus for use as live, attenuated vaccine in turkeys.

We report the development of a cold adapted strain of avian pneumovirus (APV) and its evaluation as a live vaccine candidate in 2-week-old turkey poults. A US isolate of APV (APV/MN/turkey/1-a/97) was serially passaged in Vero cells for 41 passages and then adapted to grow at sub-optimal temperatures by growing successively at 35, 33 and 31 degrees C for eight passages at each temperature. The virus thus adapted to grow at 31 degrees C was used as a candidate vaccine. The birds were vaccinated with two different doses of cold adapted virus and challenged with virulent virus 2 weeks after vaccination. No clinical signs were observed post-vaccination. Upon challenge, no clinical signs were seen in vaccinated birds but severe clinical signs were seen in non-vaccinated, challenged birds. The signs included unilateral or bilateral mucoid nasal discharge, watery eyes and swelling of infraorbital sinuses. The antibody levels in vaccinated birds were not very high. None of the vaccinated birds were found to shed virus after challenge in their choanal secretions whereas all of the non-vaccinated, challenged birds shed the virus. The absence of clinical signs and virus shedding in vaccinated birds as compared to that in non-vaccinated birds suggests that the cold adapted strain of APV is a viable candidate for use as a live, attenuated vaccine in turkeys.

Experimental and field evaluation of a live vaccine against avian pneumovirus.

The attenuation of an avian pneumovirus (APV) isolate (APV/MN/turkey/1-a/97) by 63 serial passages in cell culture (seven in chicken embryo fibroblasts and 56 in Vero cells) and its evaluation as a live attenuated vaccine in turkey poults is described. The birds were vaccinated with two different doses of attenuated virus (10(4.5) median tissue culture infectious dose (TCID(50))/ml and 10(2.5) TCID(50) /ml) at 2 weeks of age, and were challenged 2 weeks later with virulent APV. No clinical signs were seen in vaccinated, challenged birds, whereas severe clinical signs were observed in the mock-vaccinated, challenged group. Vaccinated birds developed anti-APV antibodies, which increased in titre following challenge with virulent virus. On challenge, none of the vaccinates was found to shed viral nucleic acid as detected by reverse transcriptase-polymerase chain reaction, but non-vaccinated, challenged birds did. The vaccine virus was also evaluated under field conditions in two farms. At one farm, the 'seeder bird approach' was used and two birds per 1,000 birds were vaccinated by the oculo-nasal route. In the second farm, the virus was given to all birds simultaneously in the drinking water. The birds vaccinated by the drinking water route seroconverted earlier and continued to shed virus for longer as compared with birds inoculated by the seeder bird approach. The overall results of this study indicate that the 63rd passage of APV was sufficiently attenuated and offered protection against challenge with virulent virus.

Effect of an immunomodulator on the efficacy of an attenuated vaccine against avian pneumovirus in turkeys.

Since 1997, avian pneumovirus (APV) has caused estimated annual losses of $15 million to the Minnesota turkey industry. In order to develop an attenuated live vaccine against APV, we serially passaged a Minnesota isolate of APV (APV/MN/turkey/1-a/97) in vitro in cell cultures for 41 passages. Laboratory experiments with this high-passage virus (P41) indicated that the attenuated virus provided immunogenic protection to turkeys against challenge with virulent APV, although some birds showed mild to moderate dinical signs after inoculation. To reduce the residual pathogenicity of P41, while maintaining its immunogenicity, we decided to vaccinate turkeys with P41 in the presence of an immunomodulator, S-28828 (1-n-butyl-2-ethoxymethyl-1H-imidazo[4,5-c]quinolin-4-amine-hydrochloride), which is a potent cytokine inducer. The combined inoculation of S-28828 (5 mg/kg body weight) and P41 resulted in a significant reduction in the incidence of virus-induced clinical signs in comparison with birds that received P41 without immunomodulator (P < 0.05). Only 17% of birds inoculated with S-28828 + APV P41 showed mild respiratory symptoms at 5 days postinoculation as compared with 46% of the vaccinated turkeys that did not receive S-28828. Vaccination with either P41 or with P41 + S-28828 protected turkeys against dinical signs and viral replication after challenge with virulent APV. These results indicate that immunomodulators, such as S-28828, may act as good vaccine adjuvants that can reduce the pathogenicity but maintain the immunogenicity of partially attenuated vaccines.