As the most important cultural crustacean species worldwide, studies about Pacific white shrimp (Litopenaeus vannamei) have received more attention. It has been well-documented that various pathogens could infect L. vannamei, resulting in huge economic losses. The studies about the responding mechanism of L. vannamei to sole pathogens such as Vibrio parahaemolyticus and white spot virus (WSSV) have been extensively reported, while the studies about the differently responding mechanisms remain unclear. In the present study, we identified the differently expressed genes (DEGs) of L. vannamei hemocytes post V. parahaemolyticus and WSSV infection with RNA-seq technology and compared the DEGs between the two groups. The results showed 2672 DEGs post the V. parahaemolyticus challenge (1079 up-regulated and 1593 down-regulated genes), while 1146 DEGs post the WSSV challenge (1067 up-regulated and 513 down-regulated genes). In addition, we screened the genes that simultaneously respond to WSSV and V. parahaemolyticus (434), solely respond to WSSV (1146), and V. parahaemolyticus challenge (2238), respectively. Six DEGs involved in innate immunity were quantified to validate the RNA-seq results, and the results confirmed the high consistency of both methods. Furthermore, we found plenty of innate immunity-related genes that responded to V. parahaemolyticus and WSSV infection, including pattern recognition receptors (PRRs), the proPO activating system, antimicrobial peptides (AMPs), and other immunity-related proteins. The results revealed that they were differently expressed after different pathogen challenges, demonstrating the complex and specific recognition systems involved in defending against the invasion of different pathogens in the environment. The present study improved our understanding of the molecular response of hemocytes of L. vannamei to V. parahaemolyticus and WSSV stimulation.

作为全球最重要的文化甲壳类动物，对太平洋白虾（Litopenaeus vannamei）的研究越来越受到关注。有充分证据表明，多种病原体都可能感染南美白对虾，造成巨大的经济损失。关于南美白对虾对副溶血性弧菌、白斑病毒（WSSV）等单一病原体的响应机制的研究已被广泛报道，但不同响应机制的研究尚不清楚。在本研究中，我们利用RNA-seq技术鉴定了副溶血性弧菌和WSSV感染后南美白对虾血细胞的差异表达基因（DEG） ，并比较了两组之间的DEG。结果显示，副溶血弧菌攻击后有 2672 个 DEG（1079 个上调基因和 1593 个下调基因），而 WSSV 攻击后有 1146 个 DEG（1067 个上调基因和 513 个下调基因）。此外，我们还筛选了同时响应 WSSV 和副溶血弧菌(434)、仅响应 WSSV (1146) 和副溶血弧菌攻击 (2238) 的基因。对涉及先天免疫的6个DEG进行定量以验证RNA-seq结果，结果证实了两种方法的高度一致性。此外，我们发现了大量对副溶血弧菌和 WSSV 感染有反应的先天免疫相关基因，包括模式识别受体 (PRR)、proPO 激活系统、抗菌肽 (AMP) 和其他免疫相关蛋白。结果显示，它们在不同病原体攻击后表达不同，证明了防御环境中不同病原体入侵所涉及的复杂而特异的识别系统。本研究提高了我们对南美白对虾血细胞对副溶血弧菌和 WSSV 刺激的分子反应的理解。