Preparation And Application Of Pseudomonas Aeruginosa-klebsiella Pneumoniae Bivalent Subunit Vaccine In Mink

Currently,most mink cultivation is based on scattering raising households,which have weak awareness of anti-epidemic management.High-density cultivation,pollution of stool and abuse of antibiotics result in generation and mass propagation of pseudomonas aeruginosa,Klebsiella pneumonia and other comment virulence factor-carried bacteria,and the issue of drug resistance caused by pathogenic bacteria is increasingly outstanding.In recent years,mink pneumonia caused by pseudomonas aeruginosa and Klebsiella pneumonia frequently breaks out during mink molt period and causes heavy economic losses to raising households.As vaccines available in the market are all clinically isolated intact somatic antigens with the risk of carrying drug resistance gene and virulence genes and as complicated serotypes result in uncertain immune effect,in the study,clinically isolated mink hemorrhagic pneumonia virulent strain-pseudomonas aeruginosa refined protein components and Klebsiella capsular polysaccharide were adopted as the antigen to prepare mink pseudomonas aeruginosa-Klebsiella pneumoniae bivalent subunit vaccine;the vaccine was applied for immunization of newly born cub minks in mink farm to verify the effect of the vaccine for prevention and control of mink pneumonia.In the study,conventional methods were adopted to extract and refine Klebsiella capsular polysaccharide;ion-exchange column chromatography was used to clinically extract antigen components of mink hemorrhagic pneumonia virulent strain-pseudomonas aeruginosa;with aluminum hydroxide as the adjuvant,the antigens were used to prepare vaccines.Different antigen groups were set.Challenge protection experiment was conducted in immunized mice.It is indicated that theantigen of split supernatant of pseudomonas aeruginosa with aluminum hydroxide as the adjuvant and the antigen of inactivated pseudomonas aeruginosa both induced generation of specific antibodies in immunized mice,and the two groups showed no significantdifference of antigen-induced antibody level.Namely,both antigen types showed fair protective effect in minks challenged by clinically isolated virulent strains of pseudomonas aeruginosa.Klebsiella pneumonia capsular polysaccharide showed favorable antigenicity,and induced a higher specific antibody level in mice than Klebsiella pneumonia somatic antigen,and the difference was significant(P<0.05).To verify the immunoprotection effects of the antigen types over original host minks,optimized antigen components were used to immunize minks according to immune procedure of an inactivated vaccine.It was indicated by the challenge protection experiment that the antigen of multi-component proteins refined from split supernatant of pseudomonas aeruginosa induced generation of specific antibodies in minks,and the antibody level showed no significant difference from inactivate somatic antigen of pseudomonas aeruginosa.Multi-component proteins refined from split supernatant of pseudomonas aeruginosa and Klebsiella pneumonia capsular polysaccharide antigens were used with aluminum hydroxide as the adjuvant to prepare the vaccine.The capsular polysaccharide in the prepared pseudomonas aeruginosa-Klebsiella pneumoniae bivalent subunit vaccine showed favorable antigenicity,as well as 100% protection effect for mink pneumonia caused by pseudomonas aeruginosa and Klebsiella pneumonia.The pseudomonas aeruginosa-Klebsiella pneumoniae bivalent subunit vaccine was prepared in bulk and used in a popularization and application study in mink farm.It was indicated that,during the cultivation of minks,the pseudomonas aeruginosa-Klebsiella pneumoniae bivalent subunit vaccine showed good protection effect over mink pneumonia caused by pseudomonas aeruginosa and Klebsiella pneumonia in cub minks born in the year and immunized with the vaccine.In conclusion,the bivalent subunit vaccine prepared with antigens of Klebsiella pneumonia capsular polysaccharide and pseudomonas aeruginosa-refined multiple protein components overcomes the problems of enhanced pathogenic virulence and drug resistance,favorably protects minks,and can be used as a safe,efficient and biological risk-free subunit vaccine candidate for prevention and control of pneumonia in minks.