Oral Administration of Bacterially Expressed VP28dsRNA to Protect Penaeus monodon from White Spot Syndrome Virus

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Abstract
We explored the possibility of protecting Penaeus monodon against white spot syndrome virus (WSSV) infection via interference RNA technology by oral administration of bacterially expressed WSSV VP28dsRNA. Shrimp were given dsRNA orally via two methods. In the first method, pellet feed was coated with inactivated bacteria containing overexpressed dsRNA of the WSSV VP28 gene, and in the second method, pellet feed was coated with VP28dsRNA–chitosan complex nanoparticles. The treated shrimp were orally challenged with WSSV by feeding WSSV-infected tissue. The experiment was conducted for 30 days. The dsRNA-treated shrimp challenged with WSSV showed higher survival compared to control shrimp. Sixty-eight percent survival was observed in shrimp fed with feed coated with inactivated bacteria containing dsRNA of the WSSV VP28 gene whereas 37% survival was observed in shrimp fed with VP28dsRNA–chitosan complex nanoparticle–coated feed. The WSSV caused 100% mortality in shrimp fed with pellet feed coated with inactivated bacteria with empty LITMUS38i vector. At the end of the experiment, the tissue samples prepared from randomly selected shrimp that survived were analyzed via reverse transcriptase-polymerase chain reaction and Western blot analysis for WSSV. The samples were negative for WSSV. Based on the present data and the advantages of dsRNA, we believe that oral administration of crude extract of bacterially expressed VP28dsRNA is a potential therapeutic agent against WSSV infection of shrimp.