Abstract

White spot syndrome virus (WSSV) is the most devastating virosis threatening the shrimp culture industry worldwide. Variations of environmental factors in shrimp culture ponds usually lead to the outbreak of white spot syndrome (WSS). In order to know the molecular mechanisms of WSS outbreak induced by temperature variation and the biological changes of the host at the initial stage of WSSV acute infection, RNA-Seq technology was used to analyze the differentially expressed genes (DEGs) in shrimp with a certain amount of WSSV cultured at 18°C and shrimp whose culture temperature were raised to 25°C. To analyze whether the expression changes of the DEGs were due to temperature rising or WSSV proliferation, the expression of selected DEGs was analyzed by real-time PCR with another shrimp group, namely Group T, as control. Group T didn’t suffer WSSV infection but was subjected to temperature rising in parallel. At the initial stage of WSSV acute infection, DEGs related to energy production were up-regulated, whereas most DEGs related to cell cycle and positive regulation of cell death and were down-regulated. Triose phosphate isomerase, enolase and alcohol dehydrogenase involved in glycosis were up-regulated, while pyruvate dehydrogenase, citrate synthase and isocitrate dehydrogenase with NAD as the coenzyme involved in TCA pathway were down-regulated. Also genes involved in host DNA replication, including DNA primase, DNA topoisomerase and DNA polymerase showed down-regulated expression. Several interesting genes including crustin genes, acting binding or inhibiting protein genes, a disintegrin and metalloproteinase domain-containing protein 9 (ADAM9) gene and a GRP 78 gene were also analyzed. Understanding the interactions between hosts and WSSV at the initial stage of acute infection will not only help to get a deep insight into the pathogenesis of WSSV but also provide clues for therapies.

Highlights

  • White spot syndrome (WSS) is the most devastating virosis threatening the shrimp culture industry worldwide [1]

  • Shrimp E. carincauda, cultured at 18uC, were challenged with white spot syndrome virus (WSSV), followed by temperature rising to 25uC to mimic the state transition of the infected shrimp in culture ponds as demonstrated previously [12,13]. It aims to get a better understanding of the biological changes of shrimp at the initial stage of the rapid propagation of WSSV and explore the initiation mechanisms of the acute infection caused by temperature rising

  • In Group W, no mortality appeared until 96 hptr, followed by the WSS outbreak beginning at 120 hptr

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Summary

Introduction

White spot syndrome (WSS) is the most devastating virosis threatening the shrimp culture industry worldwide [1]. Shrimp E. carincauda, cultured at 18uC, were challenged with WSSV, followed by temperature rising to 25uC to mimic the state transition of the infected shrimp in culture ponds as demonstrated previously [12,13]. It aims to get a better understanding of the biological changes of shrimp at the initial stage of the rapid propagation of WSSV and explore the initiation mechanisms of the acute infection caused by temperature rising. Cephalothorax samples from Group W and Group WT at 12 hptr were subjected to Illumina sequencing

Maintenance of Experimental Shrimp and Viral Inocula Preparation
RNA Extraction and cDNA Synthesis
Analyzing the Biological Changes of the Host from the Transcriptomic Data
Quantitative Real-time PCR Analysis of Selected Genes
Mortality and the Viral Load Variations of Group W and
Analysis of Some Interested Genes from the Transcriptomic Data
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