Cultivation And Characterization Of Goose Parvovirus In Goose Fibroblasts Cells

Gosling plague is one of the most serious infectious diseases in goose industry. Gosling could be protected by maternal antibodies from egg-laying geese injected with gosling plague vaccine (Goose parvovirus, GPV). However, gosling plague is still a problem sometimes. In this research, we isolated GPV strains by goose embryo inoculation, then these isolates of the was analyzed by SDS-PAGE and Western Blot. Two of them were passaged in goose embryo fibroblast (GEF) respectively and continuously, made them adapt GEF and attenuated, which will be made an attenuated GPV vaccine for gosling.GPV isolates which cause goose embryo dead and not agglutinate chicken red blood cells were identified by Western-blot with anti-GPV monoclonal antibody. The fourteen GPV allantoic fluids were treated with chloroform two times and concentrated ten times by PEG6000. We found that AHTC20100225strain has the highest oncentration of VP3by comparing with other GPV strains.AHTC20100225strain and GPV-SYG61strain were passaged in goose embryo fibroblast (GEF) respectively and detected by indirect immunofluorescence assay. The results showed that AHTC20100225strain adapted GEF after the fourteenth passage and GPV-SYG61strain after the tenth passage showed positive in immunofluorescent assay respectively. The TCID50of the two viruses were checked every five passages, the results showed their TCID50were all about10’5/0.2ml. The earliest time point of the70th generation of SYG61strain (SYG61-g70) virus was12hours post infection in GEF by IFA. The ELD50of SYG61-g70was about10-1.5/0.2ml, which was much lower than the ELD50of SYG61-g0(which was about10-6.0/0.2ml).One day old gosling were injected with diluted SYG61-g70, caged with normal gosling. The results showed that103TCID50of SYG61-g70couldn’t cause gosling dead. The serum neutralization test showed the antibody could be checked at5days post infection.The highest titer was10-2.84/0.2ml and maintained at about10-2/0.2ml all the time. One day old gosling immunized with diluted SYG61-g70and challaneged with JTG20100319strain, all gosling showed no symptom and health. However the mortality rate of the control group was80%. This results showed625fold of TCID50of SYG61-g70could protect the gosling injected100fold of LD50of JTG30100319strain. This virus could be an excellent vaccine virus for gosling immunization in the future.