Innate immune response of snakehead fish to Indian strain of snakehead rhabdovirus (SHRV-In) infection and the infectivity potential of the virus to other freshwater fishes.

A new virus known as snakehead rhabdovirus (SHRV-In) was discovered in South India in striped snakehead (Channa striata) that had hemorrhagic patches and cutaneous ulcerations. The virus is the most potentially harmful pathogen of snakehead because it could cause 100% mortality within 5 days. The goal of the current investigation was to evaluate the infectivity of rhabdovirus in freshwater fishes and to analyze the immune response in snakehead fish after challenge with SHRV-In. The infectivity study of SHRV-In against three freshwater fish such as tilapia, grass carp and loach showed that the virus could not induce mortality in any of them. Snakehead fish challenged with SHRV-In showed significant (p < 0.05) changes in haematological parameters such as red blood cell (RBC), haemoglobin (HGB), haematocrit (HCT), mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), white blood cell (WBC), total platelet (PLT) counts, mean platelet volume (MPV) and immunological markers such as respiratory burst, superoxide dismutase, catalase activity and myeloperoxidase activity at 6, 12, 24 and 48 hpi. Real time PCR was executed to examine the expression profile of innate immune genes such as IRF-7, IL-8 and IL-12 in Snakehead fish at 6, 12, 24 and 48 h post SHRV-In infection. Immune gene expression of IRF-7, IL-8 and IL-12 were up-regulated in the spleen when compared to kidney at 6 and 12 hpi. However, the expression level of all the genes was down-regulated at 24 and 48 hpi. The down regulation of innate immune genes after 24 hpi in these tissues may be the result of increased multiplication of SHRV-In by interfering with the immune signaling pathway.

Development of recombinant major capsid protein Vaccine and assessment of its efficacy against SRDV in similar damselfish (Pomacentrus similis).

Viral vaccines against emerging viral diseases are crucial for encouraging successful aquaculture production. In this research, an experimental recombinant major capsid protein vaccine of similar damselfish virus was prepared and examined for its efficacy in marine ornamental fish, similar damselfish (Pomacentrus similis). The MCP gene of the SRDV was amplified from the viral DNA by a specific primer set viz bamHI and XhoI- restriction sites and confirmed by agarose gel electrophoresis with a target size of 1416 bp. The gel-purified PCR product was double-digested with the said enzymes and incorporated into the pTriEx1.1 vector, which was subsequently transformed to E. coli DH5α. The plasmids of the two clones pTriEx-MCP-1416-1 and pTriEx-MCP-1416-3 were transformed to E. coli BL21 (DE-3) pLacI. A crude protein compound derived from a colony of E. coli BL21 (DE-3) with expressed MCP inserts was used to evaluate efficacy in similar damselfish by intra-peritoneal injection. After the challenge with SRDV, damselfish vaccinated with recombinant protein showed a lower protection level, while the fish vaccinated with recombinant protein supplemented Quil-A® adjuvant showed an RPS of 26%. According to RPS values recorded from the vaccinated and non-vaccinated damselfish group, the recombinant protein vaccine conferred only marginal protection against the SRDV challenge.