The Preparation And Evaluation Of Immune Efficacy Of Avian Salmonella Pullorum Ghosets

Salmonella pullorum disease (Pullorum Disease) the main cause acute systemic disease ofyoung chicken, adult chicken infection and chronic carriers rarely clinical symptomssignificantly. Currently the level of development of the poultry farming industry is relativelypoor, can not effectively control the spread of Salmonella among livestock and humans,Inactivated vaccine is often used clinically to prevent the disease, therefore imperative for aneffective vaccine.In this experiment, PhiX174phage lysis gene E and8different antimicrobial peptidesequences were inserted into the MCS1and MCS2of pre-built thermostat dual expressionvector pDUET,Construction of8different lysis plasmids. The recombinant plasmids weretransformed into salmonella. Salmonella pullorum with the recombinant plasmids were grownin a waterbath shaker at30C under aerobic conditions until mid log-phase. Then the culturewas transferred to a42°C water-bath shaker in order to make it clearage. Has the differentdegree of cracking under a scanning electron microscope, Using plate count method forcalculating the rate of bacterial lysis, to determine the best recombinant plasmid. The testanalyzes SMAP-29and E genes in pDUET have the best result, its cleavage efficiency canreach99.89%, which was named pDUET-E-SMAP29.Preparation of bacterial ghost vaccine and nactivated vaccine,the SPF chicken wereimmunized in following groups: GroupⅠ(ghost), GroupⅡ（nactivated vaccine）, GroupⅢ(PBS). The animals in each group were immunized2times with2-wk intervals. The serumantibody titers were determined using indirect enzyme-linked immunosorbent assay (iELISA)every week. CD4+and CD8+were detected by FCM. And survival rates, internal organspathological changes, the immune protection efficacy of vaccine were assessed.The results showed that ghost vacinne manifested higher protective efficacy to chickenschallenged with Salmonella pullorum through activating humoral-mediated immunity andcell-mediated immunity. In summary, Salmonella pullorum ghost vacinne have the higherprotective efficacy to chickens challenged with Salmonella pullorum.