The Molecular Mechanism of Hemocyte Immune Response in Marsupenaeus japonicus Infected With Decapod Iridescent Virus 1.

As a new type of shrimp lethal virus, decapod iridescent virus 1 (DIV1) has caused huge economic losses to shrimp farmers in China. Up to now, DIV1 has been detected in a variety of shrimps, but there is no report in Marsupenaeus japonicus. In the current study, we calculated the LC50 to evaluate the toxicity of DIV1 to M. japonicus and determined through nested PCR that M. japonicus can be the host of DIV1. Through enzyme activity study, it was found that DIV1 can inhibit the activities of superoxide dismutase, catalase, lysozyme, and phenoloxidase, which could be a way for DIV1 to achieve immune evasion. In a comprehensive study on the transcriptomic changes of M. japonicus in response to DIV1 infection, a total of 52,287 unigenes were de novo assembled, and 20,342 SSR markers associated with these unigenes were obtained. Through a comparative transcriptomic analysis, 6,900 differentially expressed genes were identified, including 3,882 upregulated genes and 3,018 downregulated genes. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that some GO terms related to virus invasion, replication, and host antiviral infection were promoted under DIV1 infection, such as carbohydrate binding, chitin binding, chitin metabolic process, and DNA replication initiation, and some KEGG pathways related to immune response were significantly influenced by DIV1 infection, including Toll and IMD signaling pathway, JAK-STAT signaling pathway, IL-17 signaling pathway, C-type lectin receptor signaling pathway, complement and coagulation cascades, antigen processing and presentation, necroptosis, apoptosis, NOD-like receptor signaling pathway, apoptosis-multiple species, and TNF signaling pathway. Further analysis showed that STAT, Dorsal, Relish, heat shock protein 70 (HSP70), C-type lectins, and caspase play an important role in DIV1 infection. This is the first detailed study of DIV1 infection in M. japonicus, which initially reveals the molecular mechanism of DIV1 infection in M. japonicus by using the transcriptome analysis of hemocytes combined with enzyme activity study.

Growth trait gene analysis of kuruma shrimp (Marsupenaeus japonicus) by transcriptome study.

Growth traits are a vital standard for the animal culture industry. The molecular mechanism of growth traits remains poorly understood, especially in aquaculture, which hinders the development of the selective breeding industry. Genomic resources discovered by next-generation sequencing (NGS) have been widely applied in certain species. However, accurate assembly and downstream analysis by NGS are still major challenges for species without reference genomes. In this study, a comparative transcriptome analysis of an economic crustacean species (Marsupenaeus japonicus) between a fast growth group and slow growth group at different stages was performed by SMRT (single molecule real time) and NGS. A high-quality full-length transcriptome (e.g., mean length of unigenes was longer than those unigenes assembled by Illumina clean reads from previous reports, and annotation rate was higher than Illumina sequencing in the same studies) was generated and analyzed. Several differentially expressed genes (DEGs) related to growth were identified and validated by quantitative real-time PCR (qPCR). The results showed that compared with the late stage, more DEGs were identified at the early stage, indicating that the growth-related physiological activity differences between different individuals at the early stage were higher than at the late stage. Moreover, 215 DEGs were shared between the early stage and late stage, and 109 had divergent functions during development. These 109 genes may play an important role in regulating the specific growth rate (SGR) of kuruma shrimp. In addition, twelve growth-related pathways were shared between the two comparative groups. Among these pathways, the fly Hippo signaling pathway and its key gene Mj14-3-3-like were identified for the first time to be involved in growth traits in crustaceans. Further analysis showed that Mj14-3-3-like was significantly downregulated in the fast growth group at the early stage and late stage; its expression level was reduced to its lowest level at the intermolt stage (C), the most important growth stage in shrimp, suggesting that Mj14-3-3-like may inhibit the growth of kuruma shrimp. Our study helps to elucidate the genes involved in the molecular mechanisms governing growth traits in kuruma shrimp, which is valuable for future shrimp developmental research.

Dietary supplementation with polypeptides improved growth performance, antibacterial immune and intestinal microbiota structure of Litopenaeus vannamei.

Antibacterial peptides (AMPs) are expected to replace some or all of the antibiotics and become a new feed additive. However, the high production cost and unclear mechanism limited the application of AMPs. In this research, the effects of a commercial polypeptide (Polypeptide S100) whose main components are AMPs on the growth, antibacterial immune and intestinal microbial of Litopenaeus vannamei were study. L. vannamei (initial weight of 0.16 ±â€¯0.03 g) were fed for 123 days with basal diet added Polypeptide S100 at two levels each (0.5% and 1%) as experimental groups, and a basal diet as control. Dietary inclusion of Polypeptide S100 at 1% level significantly increased the weight gain (WG) and specific growth rate (SGR) of L. vannamei. The survival rates of L. vannamei in 0.5% and 1% Polypeptide S100 groups were significantly higher than the control when infected by Vibrio harveyi but not Vibrio parahaemolyticus. The activities of total superoxide dismutase (T-SOD) and lysozyme (LZM) in the two experimental groups were all significantly higher than the control. Differently, the activities of amylase (AMS) and lipase (LPS) were significantly higher in 0.5% Polypeptide S100 group but lower in 1.0% Polypeptide S100 group. Illumina MiSeq high-throughput sequencing showed that the dominant phyla in the intestine of L. vannamei were Proteobacteria, followed by Actinobacteria, Bacteroidetes, Chloroflexi, Cyanobacteria, Fusobacteria and Tenericutes, and the abundance of predominant phyla Cyanobacteria were upregulated significantly in the experimental groups. At the family level, significant increase was observed in Pseudomonadaceae and Xanthomonadaceae but decrease in Vibrionaceae in the 1.0% Polypeptide S100 group. The abundance of predominant genus Photobacterium were obviously downregulated in the two experimental groups. Unlikely, the abundance of Pseudomonas and Stenotrophomonas were distinctly increased in the 1.0% Polypeptide S100 group but not significantly different from the control in 0.5% Polypeptide S100 group. All these results suggested that Polypeptide S100 could improve the growth performance, antibacterial immune and intestinal microbiota structure of L. vannamei.