A new insight to characterize immunomodulation based on hepatopancreatic transcriptome and humoral immune factor analysis of the Cherax quadricarinatus infected with Aeromonas veronii.

Cherax quadricarinatus is a type of large freshwater crayfish that is characterized by rapid growth and formidable adaptability. It has also been widely cultured and studied as a model organism. Aeromonas veronii is the dominant pathogen in aquatic environments and the primary threat to aquaculture's economic stability. To better understand the interactions between C. quadricarinatus and A. veronii, high-throughput RNA sequencing of the C. quadricarinatus hepatopancreas was carried out on a control group, susceptible group (6 h after infection), and resistant group (48 h after infection). A total of 65,850,929 genes were obtained. Compared with the control group, 2616 genes were up-regulated and 1551 genes were down-regulated in the susceptible group; while 1488 genes were up-regulated and 1712 genes were down-regulated in the resistant group. GO and KEGG analysis showed that these differentially expressed genes (DEGs) were associated with multiple immune pathways, including Toll-like receptors (TLRs), antigen processing and presentation, NOD-like receptor signaling pathway, phagosome, lysosome, JAK-STAT signaling pathway. qRT-PCR showed that infection by A. veronii changed the expression pattern of the serine proteinase inhibitor (SPI), crustacean hyperglycemic hormone (CHH), anti-lipopolysaccharide factor (ALF), and extracellular copper/zinc superoxide dismutase (SOD1), all of which were significantly higher than in the control group up to 48 h after infection. In addition, detection of superoxide dismutase (SOD), catalase (CAT), lysozyme (LZM), and phenoloxidase (PO) activity, as well as ceruloplasmin (CP) concentration at different times after infection showed diverse trends. Furthermore, pathological sections obtained 24 h after infection show lesions on the hepatopancreas and intestinal tissues caused by A. veronii. The results of this study provide a foundation for analyzing the immune mechanism of C. quadricarinatus infected with A. veronii at the transcriptional level and a theoretical basis for screening disease-resistant individuals to ensure healthy economic development of the aquatic industry.

Hepatopancreatic transcriptome analysis and humoral immune factor assays in red claw crayfish (Cherax quadricarinatus) provide insight into innate immunomodulation under Vibrio parahaemolyticus infection.

Red claw crayfish (Cherax quadricarinatus) is an economically and nutritionally important specie. We aimed to assess the immunostimulatory response to C. quadricarinatus infection with Vibrio parahaemolyticus. After determining the LD50, we infected C. quadricarinatus and examined the differential expression profiles of hepatopancreas transcriptional genes, and observed the temporal changes of hepatopancreas pathological sections and serum immunoenzymatic activities at different time points to reveal the infection mechanism of V. parahaemolyticus and the immune detoxification mechanism of the organism. The results showed that V. parahaemolyticus infection with C. quadricarinatus caused hepatopancreas injury and the immune enzyme activity of the organism changed with time delay. Transcriptome analysis of 47,338 single genes obtained by RNA sequencing and de nove transcriptome assembly identified a total of 3678 differentially expressed genes (P < 0.05) in the expression profiles of susceptible and normal animals for comparative analysis, and 2516 differentially expressed genes (P < 0.05) in the expression profiles of asymptomatic (infection-resistant) and normal animals. GO and KEGG and analyses revealed immune-related pathways under V. parahaemolyticus infection, including Vibrio cholerae infection, phagosome, lysozyme, oxidative phosphorylation, antigen processing and presentation, apoptosis, and Toll-like receptor signaling, as well as significant differences in the expression patterns of related immune genes at different times (P < 0.05). These new experimental results reveal the molecular response of the hepatopancreas to V. parahaemolyticus infection and the corresponding adaptive mechanisms, thus further revealing the pathogenesis due to bacterial infection in the aquatic environment, and providing a reference for further understanding of microbial-host interactions in aquatic systems.

Transcriptome analysis of the Macrobrachium nipponense hepatopancreas provides insights into immunoregulation under Aeromonas veronii infection.

The oriental river prawn Macrobrachium nipponense is a commercially important freshwater shrimp that is widely farmed in China. Aeromonas veronii is a conditional pathogen of farmed shrimp, which has caused huge economic losses to the industry. Therefore, there is urgency to study the host-pathogen interactions between M. nipponense and A. veronii to screen individuals with antimicrobial resistance. In this study, we examined the hepatopancreas of moribund M. nipponense infected with A. veronii and healthy individuals at both the histopathological and transcriptomic levels. We showed that A. veronii infection resulted in tubular necrosis of the M. nipponense hepatopancreas. Such changes likely affect assimilation, storage, and excretion by the hepatopancreas, which could ultimately affect the survival and growth of infected individuals. Among the 61,345 unigenes obtained through RNA sequencing and de novo transcriptome assembly, 232 were differentially expressed between the two groups. KEGG and GO analyses revealed that these differentially expressed genes were implicated in pathways, including PPAR, PI3K/AKT, and AMPK signaling. The results of this study will contribute to an analysis of the immune response of M. nipponense to A. veronii infection at the transcriptomic level. Furthermore, the RNA-seq data generated here provide an important genomic resource for research on M. nipponense in the absence of a reference genome.