Shrimp transcriptome analysis after exposure to recombinant Vibrio parahaemolyticus PirA and PirB toxins

Vibrio parahaemolyticus is a Gram-negative bacterium commonly found in marine and estuarine environments. Acute hepatopancreatic necrosis disease (AHPND) caused by this bacterium is an ongoing problem among shrimp farming industries. V. parahaemolyticus proteins PirA and PirB have been determined to be major virulence factors that induce AHPND. In this study, Pacific white shrimp (Litopenaeus vannamei) were challenged with recombinant PirA and PirB by a reverse gavage method and then at 30 m, 1, 2, 4, and 6 h time points, the hepatopancreas of five individual shrimp were removed and placed into RNA later. We conducted RNA sequencing of the hepatopancreas samples from a no PirA/B control (n = 5) and PirA/B-treated shrimp at the different time intervals (n=5). We evaluated the different gene expression patterns between the time groups to the control with a focus on identifying differences in innate immune function. Overall design: Total 60 samples from shrimp exposed to pirAB toxins at timepoints 0.5h, 1h, 2h, 4h, and 6h compared against control condition (buffer-only at 0.5h), 5 replicates from each condition.副溶血性弧菌（Vibrio parahaemolyticus）是一种常见于海洋与河口环境的革兰氏阴性菌（Gram-negative bacterium）。该菌引发的急性肝胰腺坏死病（Acute hepatopancreatic necrosis disease, AHPND）是当前虾类养殖业面临的持续性难题。现已明确，副溶血性弧菌的PirA与PirB蛋白是诱导AHPND的主要毒力因子。 本研究采用反向灌胃法（reverse gavage method），以重组PirA和PirB蛋白攻毒凡纳滨对虾（Litopenaeus vannamei，又称太平洋白虾）。分别于攻毒后30分钟、1小时、2小时、4小时及6小时采集5只个体对虾的肝胰腺，置于RNA later中保存。我们对未添加PirA/B的对照组（n=5）及各时间点PirA/B处理组（每组n=5）的肝胰腺样本开展RNA测序（RNA sequencing），通过比较各时间组与对照组的基因表达模式差异，重点探究先天免疫功能（innate immune function）相关的表达变化。 实验总体设计：本研究共包含60个样本，涵盖暴露于pirAB毒素的0.5h、1h、2h、4h、6h共5个时间点的对虾样本，以及仅用缓冲液处理的0.5h对照组样本，每组均设置5个生物学重复。