Preparation Of Inactivated Oil Emulsion Vaccine Of Goose Reovirus

Goose reovirus disease(goose reovirus GRV)is caused by reovirus goose an infectious disease,the disease is popular in many provinces in C hina in recent years,caused great economic losses to aquaculture,the great harm to raise the healthy development of goose industry in our country.In the second half of 2015,Jiangsu province,a gaggle of a goose field in liver and spleen necrosis,movement disorders as the main characteristics of infectious disease,suspected reovirus infection.For proven disease pathogen,this study will be dead tissue and serum of the goose,goose embryo yolk sac vaccination 10 age,a virus strain was isolated from allantoic fluid,virus isolates to chicken red blood cells,blood clotting activity,its ELD50 for 10 mL-3.75/0.2.Reference GenBank goose reovirus gene sequences of conservative district design specific primers was synthesized,using the primers for extraction of viral RNA rt-pcr,banding can be expanded to about 380 bp purpose,connect the purpose with recycled and pMD18-T carrier connection,restructuring carrier into DH5 alpha and e.coli in AMP + LB culture medium,the positive colonies sequencing analysis.The results showed that the cloned genes for goose reovirus sigma C gene;Biological source software analyses the isolates and waterfowl reovirus exist in the same branch,homology in 94.8% 96.5%,and the chicken reovirus homology only 39.9% to 50.5%;In the age of 6 tests back cheeper can copy the disease;The results show that the separated strains for goose reovirus,save the viruses isolated from laboratory.Will save the goose reovirus isolates the goose embryo represented three times,harvested virus allantoic fluid,8000 rpm 15 min,the centrifugal supernatant,plus double resistance.White oil will be 10,Span-80 and aluminum stearate by certain proportion mixed oil phase cooperation,disease treated by double anti venom and 2% twain-80 production of water phase,according to the oil-water ratio 2:1 mixed preparation of oil emulsion inactivated vaccine,and including appearance,emulsion type,viscosity,stability,sterile shelf life,safety,and quality inspection.Preparation of inactivated vaccines to evaluate the immune effect,only 6 days of age 90 will be cheeper were randomly divided into three groups,each group of 30,1 to 2 groups for immune,immune dose of 0.2 mL and 0.5 mL,respectively,the third group as the control group,after the immune antibody level monitoring and immune attack poison protection test,and watch the test goose morbidity and mortality.Results showed that the vaccine for milky appearance,a water-in-oil type,sterile,accord with a standard viscosity,good stability,no adverse reaction to animals,it can save more than 12 months under 4 ?;Poison attack protection experiment,depressed immune poison attack after a brief set of spirit,1 set is in two weeks in immune still cannot fully protect cheeper,immune after a week can complete protection of two groups,control group poison attack after cheeper are typical pathological changes and death;1 set of tapping poison goose cloacal swab on the third day began to detect viral nucleic acid,there are still some 12 days sample testing positive for nucleic acid,2 groups of test negative for the virus nucleic acid;C heeper the antibodies in the blood after the immune monitoring results found that 6 d after immune antibody became obvious rise,peak around 20 d,then a slightly lower but still maintain peak more than 2 weeks.The results showed that the preparation of goose reovirus inactivated vaccine safe,stable,easy to store,transport,cheeper get strong protection,after the immune antibody levels to maintain peak weeks,covering susceptible days of age.This study of goose reovirus infection disease from suspected material success in the strain isolated goose source reovirus,to lay the foundation for further study,carried out the virus preparation with the strain of the goose reovirus inactivated vaccine,the vaccine is safe and effective,can in the susceptible age cheeper to provide strong protection,for goose reovirus vaccine development to provide the basis.