Aeromonas hydrophila, which belongs to the genus Aeromonas, is a rod-shaped, facultative anaerobic bacterium that inhabits aquatic environments. Optimal growth occurs at 28°C. Some strains of A. hydrophila can cause disease in fish, amphibians, and humans (Wiki: Aeromonas Hydrophila).
Protein Name :
3-phosphoshikimate 1-carboxyvinyltransferase
Protein pI :
7.25
Protein Weight :
46562.74
Protein Length :
455
Protein Note :
catalyzes the formation of 5-O-(1-carboxyvinyl)-3-phosphoshikimate from phosphoenolpyruvate and 3-phosphoshikimate in tryptophan biosynthesis
Molecule Role Annotation :
An A. hydrophila aroA mutant is highly attenuated in trout, and a single immunization conferred significant protection from challenge with wild type A. hydrophila (Hernanz et al., 1998).
The protective antigen GAPDH from the fish pathogen A. hydrophila, encoded by the gene gapA34 (Mu et al., 2011).
e. Vector:
The Edwardsiella tarda strain EIB202 is used a a live, attenuated vaccine vector, using the promoter P(dps) as the protein-based expression system for the antigen (Mu et al., 2011).
f. Preparation
The gapA34 gene was introduced into P(dps) and transformed into live, attenuated E. tarda strain EIB202 (Mu et al., 2011).
g. Immunization Route
Intramuscular injection (i.m.)
h. Description
A. hydrophila is a common fish pathogen, so in order to create a vaccine against this pathogen, the gapA34 gene of A. hydrophila was expressed in the bacterial vector E. tarda strain EIB202, and it was shown to increase survival of vaccinated fish (Mu et al., 2011).
i.
Fish Response
Host Strain:
Turbot (Scophtalmus maximus)
Vaccine Immune Response Type:
VO_0000287
Efficacy:
Over 60% of the fish vaccinated with the WED/pUTDgap vector vaccine survived the A. hydrophila challenge (Mu et al., 2011).
Description:
The P(dps) promoter--which was chosen due to its high transcription activity-- was shown to elicit survival in turbot, meaning that it has the potential to be an important part of bacterial vaccine vector application (Mu et al., 2011) .
Vaccination Protocol:
In vaccination experiments, healthy grass carp were distributed randomly into control group and bath immunization group (150 fish per group). For bath immunization, fish were immersed in 20 mg L−1 SWCNTs-Aera and the control group was immersed in PBS for 6 h. At the end of the vaccination, grass carp were transferred to different tanks without SWCNTs-Aera for 28 d. After 28 d, 15 fish in each replicate were euthanized in the laboratory through washrag soaked with 20.0 g/m3 MS-222. Sampled fish were dissected immediately with sterile scissors. The intestine, kidney and spleen were aseptically removed from their abdominal cavity, separately, where five fish were sampled for analyzing expression of seven immune-related genes through quantitative real-time PCR (qRT-PCR) and the experiment was performed in triplicate replicates. Thereafter, the intestine of three fish (15 fish per replicate) were also collected and pooled together as described elsewhere to extract microbial DNA for deep sequencing [29]. After 2 d, the remaining fish (15 grass carp in each replicate) were intramuscularly injected with 6.5 × 104 cfu mL−1A. hydrophila, kept for 10 days and thereafter euthanized for sampling. Subsequently, the intestine was carefully excised and maintained as above, which was also for extracting microbial DNA for deep sequencing. All the samples were placed into sterile polypropylene centrifuge tubes and stored in −80 °C until DNA extraction. All experiments were performed in triplicate (Liu et al., 2015).
Host Gene Response of
CD8A
Gene Response:
The expression of CD8A was upregulated by the Aera in the intestine, kidney and spleen (Liu et al., 2015).
1. Hernanz et al., 1998: Hernanz Moral C, Flaño del Castillo E, López Fierro P, Villena Cortés A, Anguita Castillo J, Cascón Soriano A, Sánchez Salazar M, Razquín Peralta B, Naharro Carrasco G. Molecular characterization of the Aeromonas hydrophila aroA gene and potential use of an auxotrophic aroA mutant as a live attenuated vaccine. Infection and immunity. 1998; 66(5); 1813-1821. [PubMed: 9573055].
2. Liu et al., 2015: Liu L, Gong YX, Zhu B, Liu GL, Wang GX, Ling F. Effect of a new recombinant Aeromonas hydrophila vaccine on the grass carp intestinal microbiota and correlations with immunological responses. Fish & shellfish immunology. 2015; ; . [PubMed: 25862971].
3. Mu et al., 2011: Mu W, Guan L, Yan Y, Liu Q, Zhang Y. A novel in vivo inducible expression system in Edwardsiella tarda for potential application in bacterial polyvalence vaccine. Fish & shellfish immunology. 2011; 31(6); 1097-1105. [PubMed: 21964456].
