| To study the Effects of fermentation broth of Inonotus obliquus products on the immune response of SPF chicken Newcastle disease at different times.The IOFP with the immune adjuvant CpG ODN(CpG oligonucleotide)used in combination,the two adjuvants together research on the effects of the use of SPF Newcastle disease immune effect.The research results are as follows:(1)Effects of fermentation broth of Inonotus obliquus products on the immune response of SPF chicken Newcastle disease at different times120 1-day-old SPF chickens were randomly divided into 6 groups:the I-IV group was fed whole during the trial period(group I),fed for 7 days before immunization(group II),and fed for 4 days and 3 days before and after immunization(group III),each Feeding diets containing 0.8%IOFP for 4 days at 4 days(group IV),all immunized with NDV vaccine;group V was immunized with NDV vaccine and fed with commercial feed;group VI was immunized with PBS instead of vaccine,while feeding commodities A blank control group for the feed.All animals were first vacc inated with Newcastle disease virus(NDV)attenuated vaccine at 14 days of age and boosted after 21 days.Peripheral blood serum of each group was isolated at 0,7,14,21,28,35 and 42 days after the first exemption,and NDV HI antibody titer and neutralizing antibody titer were determined;Peripheral blood lymphocytes were tested for proliferation index and detection of Th1 type and Th2 type cytokine mRNA expression levels.The results showed that the addition of IOFP to the feed can effectively improve the immune effect of the vaccine,while the feeding time of group I and group II can significantly increase the body’s NDV HI antibody titer and neutralizing antibody compared with other groups.Titer,PBMCs proliferation level and Th1 type/Th2 type cytokine mRNA expression level.Combined with the feeding cost of IOFP,7 days before immunization,IOFP is the best application for oral immunopotentiators.(2)Effect of fermentation of Inonotus obliquus and CpG on immune response of SPF chicken Newcastle disease100 1-day-old SPF chickens were randomly divided into 5 groups:CpG ODN and IOFP used lonely or synchronously groups(CV,V+IOFP and CV+IOFP groups),adjuvant free group(V group)and control group(PBS group).All animals were vaccinated with Newcastle disease virus(NDV)inactivated vaccine via intramuscular route at 14 days of age,and boosted three weeks later.At 0,7,14,21,28,35 and 42days post-inoculation firstly,the temporal changes in serum Newcastle disease hemagglutination inhibition(HI)and neutralizing antibody titers were determined.Meanwhile,proliferations of peripheral blood mononuclear cells(PBMCs)isolated from each group in response to concanavalin A stimulation were detected.On days 0,21and 42 dpi,the percentages of CD4~+and CD8~+T lymphocytes and the expression levels of Th1-type(IFN-γ)and Th2-type(IL-4)cytokines were detected by flow cytometry and ELISA methods,respectively.Results showed that IOFP combined with CpG ODN didn’t demonstrate significant immunoenhancement effect of vaccine,including HI antibody titers,proliferation of PBMCs,proportions of CD4~+and CD8~+T lymphocytes and expression of Th1/Th2 cytokines,when comparing with them used lonely. |
