Preparation And Efficacy Of The Ghost Of Escherichia Coli Inducing Nephritis And Enteritis

Avian colibacillosis is caused by a group of pathogens designated avian pathogenic Escherichia coli(APEC).Despite being known for over a century,avian colibacillosis remains one of the major endemic diseases which afflicting the poultry industry worldwide.Escherichia coli inducing Nephritis and enteritis can make chicks kidney swelling and even bleeding,which is extremely rare in the clinical symptoms of E.coli,and the bacteria have a strong resistance drug to Penicillin,streptomycin and gentamicin.Researching an effective vaccines to prevent the disease is very important.Now,with the drug-resistant strains and multiple drug-resistant strains appearing,the effect of antibiotics is not as good as the previous.Although inactivated vaccine have better protective effect but there are shortcomings on the serotypes specific.So researching a new vaccines gradually become one of the focus of the researchers.Bacteria(bacterial ghost,BG)is a non-nucleic acid and other substances in the bacterial shell,only retaining the bacterial surface morphology and antigenic determinants.It has good immunogenicity and adjuvant function.Preparing high efficiency ghost of E.coli bacteria inducing Nephritis and enteritis is the purpose of this study.The method was adopted to realize lysis protein E and nuclease A gene SN series of genes through 15 flexible amino acids was fusion PCR,then fusion genes were inserted into the temperature controlled expression vector pBV220,Using this method to build plasmid pBV-ES which was pyrolysis temperature controlled double gene expression plasmid,then PCR amplification of the temperature controlled double gene cracking box into the shuttle vector pBA1100 to build temperature controlled double gene cracking shuttle plasmid pBA1100-DLS-ES,this plasmid can produce high efficiency ghost of Pasteurella multocida and Escherichia coli through the temperature control system.This study electroplate the plasmid into E.coli induced Nephritis and enteritis,temperature was shifted up to induce cracking protein expression to prepare ghost of E.coli bacteria.The results of OD600 nm and electron microscope showed that the processing cell lysis rate was up to 99.9999% after two hours of inducing.This study use the bacterial ghost formation mechanism to prepare ghost of E.coli bacteria induced Nephritis and enteritis to provide the base method for the new bacterial ghost vaccine preparation.To study the Immune protection effect of the ghost vaccine of E.coli bacteria induced Nephritis and enteritis by animal experiment.The healthy chicks were randomly divided into 4 groups,which were subcutaneous injected the vaccine which prepared in different ways: Inactivated vaccine group,single cleavage bacteria vaccine group and double cleavage bacteria vaccine group,control group.Each group of chickens was injected a dose of 0.2mL(containing bacteria is about 1 * 109 CFU / mL),the control group injected 0.2mL physiological saline,boost immune after the first immunization.Two weeks after the second immunization again,injecting the E.coli bacteria induced Nephritis and enteritis dose of 7.3*107CFU each chick through intramuscular.the effects of protection rate and serum antibody on the immunoprotection were investigated by intramuscular injection.The results showed that the protection rate of the BGs was better than that of the inactivated vaccine group.This study provides a new way to prevent and treat the drug-resistant E.coli disease.