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Generation of biotechnology-derived Flavobacterium columnare ghosts by PhiX174 gene E-mediated inactivation and the potential as vaccine candidates against infection in grass carp.

Zhu W, Yang G, Zhang Y, Yuan J, An L - J. Biomed. Biotechnol. (2012)

Bottom Line: Fish immunized with FCG showed significantly higher serum agglutination titers and bactericidal activity than fish immunized with FKC or PBS.Most importantly, after challenge with the parent strain G4, the relative percent survival (RPS) of fish in FCG group (70.9%) was significantly higher than FKC group (41.9%).These results showed that FCG could confer immune protection against F. columnare infection.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Animal Resistance Biology of Shandong, College of Life Sciences, Shandong Normal University, Jinan 250014, China.

ABSTRACT
Flavobacterium columnare is a bacterial pathogen causing high mortality rates for many freshwater fish species. Fish vaccination with a safe and effective vaccine is a potential approach for prevention and control of fish disease. Here, in order to produce bacterial ghost vaccine, a specific Flavobacterium lysis plasmid pBV-E-cat was constructed by cloning PhiX174 lysis gene E and the cat gene with the promoter of F. columnare into the prokaryotic expression vector pBV220. The plasmid was successfully electroporated into the strain F. columnare G4cpN22 after curing of its endogenous plasmid. F. columnare G4cpN22 ghosts (FCGs) were generated for the first time by gene E-mediated lysis, and the vaccine potential of FCG was investigated in grass carp (Ctenopharyngodon idellus) by intraperitoneal route. Fish immunized with FCG showed significantly higher serum agglutination titers and bactericidal activity than fish immunized with FKC or PBS. Most importantly, after challenge with the parent strain G4, the relative percent survival (RPS) of fish in FCG group (70.9%) was significantly higher than FKC group (41.9%). These results showed that FCG could confer immune protection against F. columnare infection. As a nonliving whole cell envelope preparation, FCG may provide an ideal alternative to pathogen-based vaccines against columnaris in aquaculture.

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Serum agglutination titres of fish vaccinated with FCG and FKC at different time points postprimary immunization. Grass carp were vaccinated with FCG or FKC via intraperitoneal injection with a boost at 4 weeks after primary immunization. Serum agglutination titres represent the highest dilutions that gave rise to positive reaction. The titres of FCG group were significantly higher than FKC group at all the examined time points (P < 0.01).
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fig4: Serum agglutination titres of fish vaccinated with FCG and FKC at different time points postprimary immunization. Grass carp were vaccinated with FCG or FKC via intraperitoneal injection with a boost at 4 weeks after primary immunization. Serum agglutination titres represent the highest dilutions that gave rise to positive reaction. The titres of FCG group were significantly higher than FKC group at all the examined time points (P < 0.01).

Mentions: The agglutination reaction against F. columnare G4 was detected in grass carp immunized ip with bacterial ghosts, formalin-killed cells, or PBS. Fish immunized with FCG or FKC showed significantly higher agglutination titer than control fish in which no agglutination reaction was detected, while fish immunized with FCG showed significantly higher titers than fish immunized with FKC at all the examined time points (P < 0.01) (Figure 4). The agglutination reaction in FCG group initially peaked at 2 weeks postprimary immunization, subsided by week 4 and then more than doubled (mean geometric titre 478) 2 weeks after booster immunization (6 weeks). The high titer of FCG group was 3.5-fold higher than FKC group and persisted longer than FKC group.


Generation of biotechnology-derived Flavobacterium columnare ghosts by PhiX174 gene E-mediated inactivation and the potential as vaccine candidates against infection in grass carp.

Zhu W, Yang G, Zhang Y, Yuan J, An L - J. Biomed. Biotechnol. (2012)

Serum agglutination titres of fish vaccinated with FCG and FKC at different time points postprimary immunization. Grass carp were vaccinated with FCG or FKC via intraperitoneal injection with a boost at 4 weeks after primary immunization. Serum agglutination titres represent the highest dilutions that gave rise to positive reaction. The titres of FCG group were significantly higher than FKC group at all the examined time points (P < 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3376489&req=5

fig4: Serum agglutination titres of fish vaccinated with FCG and FKC at different time points postprimary immunization. Grass carp were vaccinated with FCG or FKC via intraperitoneal injection with a boost at 4 weeks after primary immunization. Serum agglutination titres represent the highest dilutions that gave rise to positive reaction. The titres of FCG group were significantly higher than FKC group at all the examined time points (P < 0.01).
Mentions: The agglutination reaction against F. columnare G4 was detected in grass carp immunized ip with bacterial ghosts, formalin-killed cells, or PBS. Fish immunized with FCG or FKC showed significantly higher agglutination titer than control fish in which no agglutination reaction was detected, while fish immunized with FCG showed significantly higher titers than fish immunized with FKC at all the examined time points (P < 0.01) (Figure 4). The agglutination reaction in FCG group initially peaked at 2 weeks postprimary immunization, subsided by week 4 and then more than doubled (mean geometric titre 478) 2 weeks after booster immunization (6 weeks). The high titer of FCG group was 3.5-fold higher than FKC group and persisted longer than FKC group.

Bottom Line: Fish immunized with FCG showed significantly higher serum agglutination titers and bactericidal activity than fish immunized with FKC or PBS.Most importantly, after challenge with the parent strain G4, the relative percent survival (RPS) of fish in FCG group (70.9%) was significantly higher than FKC group (41.9%).These results showed that FCG could confer immune protection against F. columnare infection.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Animal Resistance Biology of Shandong, College of Life Sciences, Shandong Normal University, Jinan 250014, China.

ABSTRACT
Flavobacterium columnare is a bacterial pathogen causing high mortality rates for many freshwater fish species. Fish vaccination with a safe and effective vaccine is a potential approach for prevention and control of fish disease. Here, in order to produce bacterial ghost vaccine, a specific Flavobacterium lysis plasmid pBV-E-cat was constructed by cloning PhiX174 lysis gene E and the cat gene with the promoter of F. columnare into the prokaryotic expression vector pBV220. The plasmid was successfully electroporated into the strain F. columnare G4cpN22 after curing of its endogenous plasmid. F. columnare G4cpN22 ghosts (FCGs) were generated for the first time by gene E-mediated lysis, and the vaccine potential of FCG was investigated in grass carp (Ctenopharyngodon idellus) by intraperitoneal route. Fish immunized with FCG showed significantly higher serum agglutination titers and bactericidal activity than fish immunized with FKC or PBS. Most importantly, after challenge with the parent strain G4, the relative percent survival (RPS) of fish in FCG group (70.9%) was significantly higher than FKC group (41.9%). These results showed that FCG could confer immune protection against F. columnare infection. As a nonliving whole cell envelope preparation, FCG may provide an ideal alternative to pathogen-based vaccines against columnaris in aquaculture.

Show MeSH
Related in: MedlinePlus