Histological, Physiological and Transcriptomic Analysis Reveal the Acute Alkalinity Stress of the Gill and Hepatopancreas of Litopenaeus vannamei.

The pacific white shrimp (Litopenaeus vannamei) has gradually become a promising economic species in the development of saline-alkali water fishery. The study related to the stress reaction of pacific white shrimp under alkalinity stress is still limited, which is also a critical limiting factor for its saline-alkaline aquaculture. In this study, we aim to analyse the stress reaction of pacific white shrimp under acute alkalinity stress between control group (alkalinity:40 mg/L) and treatment group (alkalinity:350 mg/L) through histological observation, physiological determination and transcriptome. In the present study, during the process of acute alkalinity stress, the activities of Na+-K+-ATPase, carbonic anhydrase, sodium/hydrogen exchanger in gill related to homeostasis were significantly changed, the activities of superoxide dismutase and catalase related to antioxidant were decreased in both gill and hepatopancreas, and the activities of protease, lipase and amylase in hepatopancreas were decreased. At the same time, different degrees of histological damages were occured in the gill and hepatopancreas under acute alkalinity stress. There were 194 and 236 different expressed genes identified in gill and hepatopancreas respectively. Functional enrichment assessment indicated that the alkalinity stress-related genes in both gill and hepatopancreas were primarily involved in fatty acid metabolism, glycolysis/gluconeogenesis, glycerophospholipid metabolism. The results indicated that the functions of homeostasis regulation, antioxidation and digestion of pacific white shrimp were decreased under acute alkalinity stress, at the same time, the energy metabolism in gill and hepatopancreas were modified to cope with alkalinity stress. This work provides important clues for understanding the response mechanism of pacific white shrimp under acute alkalinity stress.

Integrated analysis of immune parameters, miRNA-mRNA interaction, and immune genes expression in the liver of rainbow trout following infectious hematopoietic necrosis virus infection.

Rainbow trout (Oncorhynchus mykiss) is an important economical cold-water fish worldwide. However, infection with infectious hematopoietic necrosis virus (IHNV) has severely restricted the development of aquaculture and caused huge economic losses. Currently, little is known about the immune defense mechanisms of rainbow trout against IHNV. In this study, we detected the changes of immune parameters over different post-infection periods (6-, 12-, 24-, 48-, 72-, 96-, 120-, and 144 hours post-infection (hpi)), mRNA and miRNA expression profiles under 48 hpi (T48L) compared to control (C48L), and key immune-related genes expression patterns in rainbow trout liver following IHNV challenge through biochemical methods, RNA-seq, and qRT-PCR, and the function of miR-330-y was verified by overexpression and silencing in vitro and in vivo. The results revealed that alkaline phosphatase (AKP), alanine aminotransferase (ALT), catalase (CAT), and total superoxide dismutase (T-SOD) activities, and lysozyme (LZM) content showed significant peaks at 48 hpi, whereas malondialdehyde (MDA) content and aspartate aminotransferase (AST) activity decreased continuously during infection, and acid phosphatase (ACP) activity varied slightly. From RNA-seq, a total of 6844 genes and 86 miRNAs were differentially expressed, and numerous immune-related differentially expressed genes (DEGs) involved in RIG-I-like receptor signaling pathway, Toll-like receptor signaling pathway, NOD-like receptor signaling pathway, cytokine-cytokine receptor interaction, and antigen processing and presentation were significantly upregulated in T48Lm group, including IFIH1, DHX58, MAVS, TRAF3, IRF3, IRF7, MX1, TLR3, TLR8, MYD88, NOD1, NOD2, IL-8, CXCR1, CD209, CD83, and TAP1. Integrated analysis identified seven miRNAs (miR-425-x, miR-185-x, miR-338-x, miR-330-y, miR-361-x, miR-505-y, and miR-191-x) that target at least three key immune-related DEGs. Expression analysis showed that IFIH1, DHX58, IRF3, IRF7, MX1, TLR3, TLR8, and MYD88 showed a marked increase after 24 hpi during infection. Further research confirmed TAP1 as one of the targets of miR-330-y, overexpression of miR-330-y with mimics or agomir significantly reduced the expression levels of TAP1, IRF3, and IFN, and the opposite effects were obtained by inhibitor. These results facilitate in-depth understanding of the immune mechanisms in rainbow trout against IHNV.