The Immunogenicity Analysis On Whole Cell Of The Haemophilus Parasuis

Haemophilus parasuis (Haemophilus parasuis, HPS),belonging to members of the genus Haemophilus,Pasteurella families,is considered to be the major pathogen of Glaser’s disease (Glaesser’s Disease). As the world’s pig industry, Haemophilus parasuis has joined the ranks of typical bacterial diseases threatening the global pig industry, but now the majority of current practice in the production is still using the traditional inactivated vaccine to prevent the disease. This research is mainly about the growth curve drawing of the epidemical serum type5parasuis in Sichuan, the determination of fully lethal dose in mice as well as the analysis inactivated whole-cell vaccine immunogenicity, provides a basis for serum type5parasuis whole-cell vaccine to use in the production; It also provides specific steps to verify the effectiveness of the vaccine, has some reference value to.the production of own vaccine and effectiveness testing.1.The measure of parasuis growth curve and fully lethal dose in mice The trial cultured the serum type5parasuis and regularly measured its OD600values,and draw out not only its growth curve,but also the OD600of correlation with the number of viable cells curve. The results show that when parasuis is in lag phase and logarithmic phase, OD600values is with a nearly linear relationship between the number of viable cells, its relevance can be represented with this formula:y=13.30x+0.385, R2=0.987the correlation can be y=13.30x+0.385, R2=0.987This formula. With the number of viable cells was respectively2.48×108CFU,2.48×109CFU,2.48×1010CFU,2.48×1011CFU of HPS in mice by intraperitoneal injection attack drug and observed number of deaths recorded shows, the parasuis content of2.48x1010CFU viable and above can make up to100%mortality in mice.2.The immunogenicity of parasuis whole-cell vaccine In this experiment, inactivated vaccine was prepared with serum type5parasuis, and the inactivated vaccine was used to carry out the related research between the vaccine antigen content and experimental animal protective immune poisoning,the related research of serological effectiveness inspection and laboratory animal protective immune poisoning, different Vaccination experiments on the effect of experimental animals,the cross protection between different serotypes, the effect that offspring obtained passive immunity through maternal antibodies, and efficacy testing laboratory animal test results with the target animal efficacy studies and other experimental correlation, to analyze the immunogenicity of the vaccine. The results showed that mice inoculated with3.24×1010CFU,3.24x109CFU,3.24×108CFU,3.24×107CFU4antigenic content of vaccine protection rate suffered by serum antibodies produced its OD value was a certain linear relationship:y=165.8x-12.77, R2=0.943, i.e. for the four inoculated with the vaccine antigen content increases, the increased levels of serum antibody, virus attack protection rate increases. When the antigen content of about3.24x109CFU above can make the mice get better protection force (protection rate of80%to100%). The results also show that the vaccine serotypes4virulent virus attack protection in mice was12.5%, indicating that the vaccine has a certain cross-immunity. The mice were inoculated in the muscle, subcutaneous and intraperitoneal inoculation, inoculating intraperitoneally has the highest level of antibodies, this result indicates that inoculating intraperitoneally vaccinated is the best way for the test mice. Target animals pigs and experimental animals mice produced antibodies are basically the same trend, they are both started one week after immunization, antibody levels gradually increased, in3to4weeks after immunization antibody levels produce better, in the first four weeks after immunization antibody levels occurs around the peak, then with time show a downward trend. While pregnant sows after vaccination in prenatal birth of piglets with a certain level of antibodies, then through breastfeeding, will greatly improve the level of antibodies, but with time, the antibody levels will gradually decrease. From the experimental results,14days after the piglets’ birth they must recieve the first immunization for the vaccine. The second vaccines on animals free time should be free in the first three weeks after the first.