Silencing of EhAQP1, EhPTP2, and EhTK Genes in Ecytonucleospora Hepatopenaei Using RNA Interference: A Potential Strategy for Preventing and Treating EHP Infection in Shrimp

Abstract

Ecytonucleospora hepatopenaei (EHP) is an obligate intracellular parasitic pathogen of shrimp, causing growth retardation and increased susceptibility to opportunistic infections. Currently, there are few reports on the prevention and treatment of EHP infection in shrimp, and research on its drug targets is limited. This article utilizes RNA interference (RNAi) technology to conduct knockdown verification studies on potential drug targets involved in EHP’s invasion mechanism and life cycle. Three target gene sequences from genome, and annotated as EhAQP1 (aquaporin 1), EhPTP2 (polar tube protein), and EhTK (thymidine kinase) were used to synthesize specific small interfering RNA (siRNA). These siRNAs were injected into EHP-infected shrimp, and the hepatopancreas was sampled at 12, 24, 48, and 72 h postinjection to verify the interference effect and spore load. The results showed that RNAi exhibits varying inhibitory effects on different genes and is time-dependent. For EhAQP1, interference was most effective in the early stages (12 to 24 h), while the interference effects for EhTK were more pronounced at 48 and 72 h, the expression of EhPTP2 was significantly downregulated (p < 0.001) only at 24 h. In the EhAQP1 and EhPTP2 interference group, the spore load decreased from 12 to 48 h, but increased at 72 h. In contrast, the EhTK interference group showed a continuous decrease in spore load from 12 to 72 h. In summary, this study provides a new strategy for prevention and treatment of shrimp EHP infection and lays the foundation for subsequent research on RNAi–based prevention and treatment of EHP infection.