Kuruma Shrimp Marsupenaeus Japonicus Research Articles

Ammonia stress-induced heat shock factor 1 enhances white spot syndrome virus infection by targeting the interferon-like system in shrimp.

Ammonia is the end product of protein catabolism and is derived from feces and unconsumed foods. It threatens the health and growth of aquatic animals. In this study, we demonstrated that ammonia stress suppresses shrimp antiviral immunity by targeting the shrimp interferon-like system and that heat shock factor 1 (Hsf1) is a central regulator of this process. When shrimp are stressed by ammonia, they activate Hsf1 for stress relief and well-being. Hsf1 upregulates the expression of negative regulators that inhibit the production and function of interferon analogs in shrimp, thereby enhancing white spot syndrome viral infection. Therefore, this study, from a molecular perspective, explains the problem in the aquaculture industry that animals living in stressed environments are more susceptible to pathogens than those living in unstressed conditions. Moreover, this study provides new insights into the side effects of heat shock responses and highlights the complexity of achieving cellular homeostasis under stressful conditions.

White spot syndrome virus hijacks host PP2A-FOXO axes to promote its propagation

Viruses have developed superior strategies to escape host defenses or exploit host components and enable their infection. The forkhead box transcription factor O family proteins (FOXOs) are reportedly utilized by human cytomegalovirus during their reactivation in mammals, but if FOXOs are exploited by viruses during their infection remains unclear. In the present study, we found that the FOXO of kuruma shrimp (Marsupenaeus japonicus) was hijacked by white spot syndrome virus (WSSV) during infection. Mechanistically, the expression of leucine carboxyl methyl transferase 1 (LCMT1) was up-regulated during the early stages of WSSV infection, which activated the protein phosphatase 2A (PP2A) by methylation, leading to dephosphorylation of FOXO and translocation into the nucleus. The FOXO directly promoted transcription of the immediate early gene, wsv079 of WSSV, which functioned as a transcriptional activator to initiate the expression of viral early and late genes. Thus, WSSV utilized the host LCMT1-PP2A-FOXO axis to promote its replication during the early infection stage. We also found that, during the late stages of WSSV infection, the envelope protein of WSSV (VP26) promoted PP2A activity by directly binding to FOXO and the regulatory subunit of PP2A (B55), which further facilitated FOXO dephosphorylation and WSSV replication via the VP26-PP2A-FOXO axis in shrimp. Overall, this study reveals novel viral strategies by which WSSV hijacks host LCMT1-PP2A-FOXO or VP26-PP2A-FOXO axes to promote its propagation, and provides clinical targets for WSSV control in shrimp aquaculture.

Mindin Activates Autophagy for Lipid Utilization and Facilitates White Spot Syndrome Virus Infection in Shrimp.

Mindin is a secreted extracellular matrix protein that is involved in regulating cellular events through interacting with integrin. Studies have demonstrated its role in host immunity, including phagocytosis, cell migration, and cytokine production. However, the function of Mindin in the host-virus interaction is largely unknown. In the present study, we report that Mindin facilitates virus infection by activating lipid utilization in an arthropod, kuruma shrimp (Marsupenaeus japonicus). Shrimp Mindin facilitates white spot syndrome virus infection by facilitating viral entry and replication. By activating autophagy, Mindin induces lipid droplet consumption, the hydrolysis of triglycerides into free fatty acids, and ATP production, ultimately providing energy for virus infection. Moreover, integrin is essential for Mindin-mediated autophagy and lipid utilization. Therefore, by revealing the mechanism by which Mindin facilitates virus infection through regulating lipid metabolism, the present study reveals the significance of Mindin in the host-virus interaction. IMPORTANCE White spot syndrome virus (WSSV) is an enveloped double-stranded DNA virus that has had a serious influence on worldwide shrimp farming in the last 30 years. We have demonstrated that WSSV hijacks host autophagy and lipid metabolism for reproduction in kuruma shrimp (Marsupenaeus japonicus). These findings revealed the mechanism by which WSSV exploits host machinery for its infection and provided serial targets for WSSV prevention and control in shrimp farming.

Preparation, Purification and Characterization of Antibacterial and ACE Inhibitory Peptides from Head Protein Hydrolysate of Kuruma Shrimp, Marsupenaeus japonicus.

Kuruma shrimp (Marsupenaeus japonicus) heads, as the main by-product of the seafood processing industry, are rich in underutilized high-quality protein. After papain hydrolysis at 50 °C for 4 h, the protein hydrolysate of shrimp heads was found to show notable antibacterial and angiotensin I-converting enzyme (ACE) inhibitory activities. After purification using two stages of revered-phase high-performance liquid chromatography (RP-HPLC), the antibacterial peptide VTVP and the ACE inhibitory peptide ARL/I were successfully identified from most active fractions by LC-MS/MS. Peptide VTVP was a desirable hydrophobic peptide, with a MIC value in the range from 1.62 to 8.03 mM against all tested pathogens. Peptide ARL/I exhibited potent ACE inhibitory activity, with an IC50 value of 125.58 µM, and was found to be a competitive inhibitor based on the Lineweaver-Burk plot. Moreover, the result of the molecular docking simulation indicated that the interaction binding between ARL/I and ACE was mainly stabilized by hydrogen bonds, as well as forming a coordinate bond with the Zn2+ site. The purified peptides did not show hemolytic activity toward rabbit erythrocytes. To sum up, the bioactive peptides isolated from shrimp heads could be applicable for food or pharmaceutical areas as promising ingredients.

Cloning of glutamine synthetase gene from abdominal muscle of kuruma shrimp Marsupenaeus japonicus and its expression profile

Cloning of glutamine synthetase gene from abdominal muscle of kuruma shrimp Marsupenaeus japonicus and its expression profile

How Do Gene Expression Patterns Change in Response to Osmotic Stresses in Kuruma Shrimp (Marsupenaeus japonicus)?

Euryhaline crustaceans cope with external salinity changes by mechanisms of osmoregulation. In the current study, we first cloned and confirmed the ORF sequences of the ion-transportation-related genes Na+/K+-ATPase α subunit (NKAα), cytoplasmic carbonic anhydrase (CAc), and V-type H+-ATPase G subunit (VHA-G), and water channels of aquaporins (AQP3, AQP4, and AQP11) from kuruma shrimp (Marsupenaeus japonicus). Further tissue expression patterns showed a higher expression of MjAQP4, MjCAc, MjNKAα, and MjVHA-G in the gills, as well as a higher expression of MjAQP3 and MjAQP11 in the intestine and muscle, respectively. Then, qPCR analysis was used to assess the mRNA expression levels of those osmoregulatory genes in both post-larvae and adult shrimp when they were exposed to acute salinity stress or salinity acclimation. The results revealed significantly decreased expression levels of MjAQP3, MjAQP11, MjNKAα, and MjCAc, and higher expression levels of MjAQP4 and MjVHA-G when the post-larvae shrimp were directly subjected to 10‰ or 50‰ salinity. Moreover, similar expression patterns were also observed in the post-larvae shrimp during the accommodation to 10‰ or 50‰ salinity. As to the adult shrimp, significantly higher expression levels of those genes were observed in the gills after exposure to 10‰ salinity, whereas only the expression levels of MjAQP3, MjAQP11, and MjNKAα were up-regulated in the gills at 40‰ salinity. In contrast, the expression of MjVHA-G was significantly decreased at 40‰ salinity. Finally, during the acclimation to 10‰ salinity, the expression levels of MjAQP3, MjAQP11, and MjNKAα were also significantly elevated, while the expression of MjCAc was significantly decreased in the gills. In addition, the expression levels of MjAQP3, MjAQP4, MjCAc, and MjVHA-G were significantly decreased in the gills during the acclimation to 55‰ salinity. The findings of the study suggest that the examined genes are critical for the adaptation of aquatic crustaceans to changing environmental salinity. Our study lays as the foundation for further research on osmoregulation mechanisms in M. japonicus.

The sORF-Encoded Peptides, ATP Synthase Subunits, Facilitate WSSV Duplication in Shrimp.

Short open reading frames (sORFs) are a newly identified family of genes, and the functions of most sORF genes and their encoded peptides (SEPs) are still unknown. In this study, two ATP synthase subunits were identified in kuruma shrimp (Marsupenaeus japonicus) as SEPs, namely MjATP5I and MjATP5L. They were widely distributed in all of the tested tissues of shrimp and upregulated in hemocytes and intestines in response to WSSV challenge. The injection of recombinant proteins (rMjATP5I and rMjATP5L) increased the expression of Ie1 and Vp28, while the knockdown of MjATP5I and MjATP5L decreased the expression of Ie1 and Vp28. All of the results suggest that MjATP5I and MjATP5L were beneficial for WSSV replication. Further exploration found that MjATP5I and MjATP5L RNAi significantly improved the shrimp survival rates, reduced ATP production, and upregulated the expression of antimicrobial peptide genes post viral challenge, and the two ATPase subunits and Relish negatively regulated each other. These results reveal that MjATP5I and MjATP5L facilitated WSSV duplication by regulating the production of ATP contents and the expression of antimicrobial peptide genes in shrimp.

Immunostimulation of shrimp through oral administration of silkworm pupae expressing VP15 against WSSV

White spot syndrome virus (WSSV) is one of the most concerning pathogens in penaeid shrimp and can cause severe loss in shrimp aquaculture worldwide. Among the WSSV structural proteins, VP15, a DNA-binding protein located in the WSSV nucleocapsid, is an antiviral protein candidate to protect kuruma shrimp (Marsupenaeus japonicus) from WSSV infection. We identified that the truncated VP15, VP15(26–57), is responsible for the protective effect against the WSSV. This study attempts to develop an immunizing agent against WSSV using silkworm pupa as a delivery vector through oral administration. The VP15, VP15(26–57), and SR11 peptide derived from VP15(26–57) were expressed in silkworm pupae. Oral administration of feed mixed with the powdered pupae that expressed VP15-derived constructs enhanced the survivability of kuruma shrimp with an overall relative percent survival (RPS) higher than 70%. There is no death for the group receiving pupa/VP15(26–57), and the RPS is 100%. In addition, we also investigated the relative mRNA expression levels of immune-related genes by qPCR at different time points. Our results indicate that the oral administration of pupa/VP15-derived products could provide a high protective effect against WSSV and be a practical approach for controlling WSSV in aquaculture.

Phenotypic and genomic characterization of pathogenic Providencia rettgeri from kuruma shrimp Marsupenaeus japonicus.

Providencia rettgeri has been recognized as a zoonotic pathogen of humans and aquaculture animals and has become a global public health concern. However, scarce information is available on the characterization of pathogenic P. rettgeri from kuruma shrimp Marsupenaeus japonicus. In the present study, a P. rettgeri isolate (KM4) was confirmed as a causative agent of red leg disease in cultured M. japonicus, which showed a median lethal dose (LD50 ) value of 5.01 × 105 CFU·ml-1 and had multiple resistances to aminoglycosides, sulfonamides, and tetracycline antimicrobials used in aquaculture. In addition, the whole genome of isolate KM4 was sequenced and found to consist of a single circular chromosome of 4,378,712bp and a circular plasmid of 171,394bp. The genome sequence analysis further revealed the presence of potential virulence and antibiotic resistance genes in isolate KM4, which probably rendered this isolate particularly virulent. To our knowledge, this is the first study to characterize P. rettgeri pathogens from kuruma shrimp infected with red leg disease. The findings of this study can provide novel insights into the presence and distribution of pathogenicity-associated genes in shrimp-pathogenic P. rettgeri.

Correction to: Isolation and molecular characterization of hemocyte sub-populations in kuruma shrimp Marsupenaeus japonicus

Correction to: Isolation and molecular characterization of hemocyte sub-populations in kuruma shrimp Marsupenaeus japonicus

Infection with white spot syndrome virus affects the microbiota in the stomachs and intestines of kuruma shrimp

Maintaining eubiosis of the gastrointestinal (GI) microbiota is essential for animal health. White spot syndrome virus (WSSV) is the most lethal viral pathogen because it causes extremely high mortality in shrimp farming. However, it remains poorly understood how WSSV infection affects the microbiota in different regions of the GI tract of shrimp. In the present study, we established an experimental model of kuruma shrimp (Marsupenaeus japonicus) infection with WSSV and then investigated the effects of WSSV infection on the microbiota in the cardiac stomach, pyloric stomach, and intestines using metataxonomics. We identified 34 phyla and 576 genera of bacteria collectively. At the phylum level, Proteobacteria and Firmicutes were the most abundant in all the three GI segments. The WSSV infection decreased microbial diversity to a different extent in the stomachs and in a time-dependent manner. The infection with WSSV affected the microbiota composition in the two stomachs, but not the intestines. Firmicutes increased significantly, while Actinobacteria, Bacteroidetes, and Cyanobacteria decreased in the two stomachs of the WSSV-infected shrimp. At the genus level, Trichococcus and Vibrio increased, but Bradyrhizobium and Roseburia decreased in the cardiac stomach of the WSSV-infected shrimp. Trichococcus and Photobacterium increased in the pyloric stomach. Although Vibrio showed a slight downward trend, Aliivibrio (formerly Vibrio) increased in the pyloric stomach. Thiothrix, Fusibacter, and Shewanella decreased in the pyloric stomach, but no significant differences in these genera were detected in the cardiac stomach. Analysis of the predicted functions of the GI microbiota indicated that the WSSV infection resulted in losses of some microbiota functions. The new information from this study may help better understand the bacteria-virus interaction in the GI tract of shrimp and other crustacean species, and inform pathogen prevention/control and sustainable aquaculture production.

A chromosome-level genome of the kuruma shrimp (Marsupenaeus japonicus) provides insights into its evolution and cold-resistance mechanism

Marsupenaeus japonicus is an important marine crustacean species. However, a lack of genomic resources hinders the use of whole genome sequencing to explore their genetic basis and molecular mechanisms for genome-assisted breeding. Consequently, we determined the chromosome-level genome of M. japonicus. Here we determine the chromosome-level genome assembly for M. japonicus with a total of 665.19 Gb genomic sequencing data, yielding an approximately1.54 Gb assembly with a contig N50 size of 229.97 kb and a scaffold N50 size of 38.27 Mb. With the high-throughput chromosome conformation capture (Hi-C) technology, we anchored 18,019 contigs onto 42 pseudo-chromosomes, accounting for 99.40% of the total genome assembly. Analysis of the present M. japonicus genome revealed 24,317 protein-coding genes and a high proportion of repetitive sequences (61.56%). The high-quality genome assembly enabled the identification of genes associated with cold-stress and cold tolerance in kuruma shrimp through the comparison of eyestalk transcriptomes between the low temperature-stressed shrimp (10 °C) and normal temperature shrimp (28 °C). The genome assembly presented here could be useful in future studies to reveal the molecular mechanisms of M. japonicus in response to low temperature stress and the molecular assisted breeding of M. japonicus in low temperature.

Filter-Feeding Bivalve Weakens Food Competition Between Crustaceans (Portunus trituberculatus, Marsupenaeus japonicus) in Integrated Culture Ponds: Evidence From 18S rDNA Barcoding and Stable Isotope Analysis

In order to investigate the effects of razor clams (Sinonovacula constricta) on the food composition and isotopic niches of swimming crabs (Portunus trituberculatus) and kuruma shrimp (Marsupenaeus japonicus) in polyculture systems, this study analyzed 60 P. trituberculatus, 60 M. japonicus and 30 S. constricta to quantify the food sources, food source contributions, and isotopic niches of cultured organisms using 18S rDNA barcoding and stable isotope techniques. The results were as follows: (1) In the P. trituberculatus-M. japonicus (PM) polyculture system, the Sobs and Shannon-Wiener indices of the stomach contents of P. trituberculatus and M. japonicus were not significantly different (P > 0.05). In the P. trituberculatus-M. japonicus-S. constricta polyculture (PMS) system, the Sobs and Shannon-Wiener indices of the stomach contents of P. trituberculatus and M. japonicus were also not significantly different (P > 0.05), but the Sobs indices of P. trituberculatus in the PMS system were significantly higher than those in the PM system (P < 0.05), M. japonicus shows a similar pattern. (2) 18S rDNA barcoding analysis showed the dominant taxa in the stomach contents of both P. trituberculatus and M. japonicus in the PM system were Trebouxiophyceae, Embryophyta and Rotifera, and the food overlap between them was 0.8992, which was significant (Q > 0.6). In the PMS system, the dominant taxa in the stomach contents of P. trituberculatus were Chrysophyceae, Intramacronucleata, and Embryophyta, and in M. japonicus were Chrysophyceae, Embryophyta, and Bacillariophyceae, in this system the food overlap was 0.2061, which was not significant (Q < 0.6). (3) Stable isotope analysis suggested, in both systems, the main food sources of P. trituberculatus and M. japonicus were iced trash fish, zooplankton, phytoplankton, and organic particulate matter (POM). Iced trash fish accounted 77.67% of food sources for P. trituberculatus and 69.42% for M. japonicus in the PM system, and 60.82% and 57.60% in the PMS system. (4) The isotopic niche overlap between P. trituberculatus and M. japonicus was 5.69% in the PM system and 1.21% in the PMS system. These results suggested food competition between P. trituberculatus and M. japonicus, and S. constricta can reduce the competition and isotopic niche overlap, improve the contribution of food sources such as phytoplankton. Razor clams also serve to purify the water and improve the utilization of iced trash fish by filtering phytoplankton (51.10%), POM (32.25%), SOM (7.47%), and iced trash fish (9.18%). Thus, P. trituberculatus-M. japonicus-S. constricta is a healthy and sustainable culture model.

Comparison of the probiotic effects of freshwater and marine photosynthetic bacteria in kuruma shrimp (Marsupenaeus japonicus) culture

ABSTRACT In a field study, the probiotic effects of a freshwater purple nonsulfur photosynthetic bacterium (PNSB), Rhodobacter sphaeroides, and a marine PNSB, Rhodovulum sulfidophilum, on kuruma shrimp (Marsupenaeus japonicus) were compared by using two commercially operating shrimp ponds. Shrimp were fed with feed containing 0.01% fresh weight (106 cfu/g feed) of PNSB cells for 152 d. The marine PNSB showed a better performance than the freshwater PNSB in average body weight at harvest time, survival rate (P < 0.001 by Fisher’s exact test), and feed conversion rate. The effects of freshwater and marine PNSB on shrimp gene expression were also compared by next-generation sequencing and quantitative RT-PCR. Both freshwater and marine PNSB promoted the expression of several genes related to the innate immune system, including NADPH oxidase, superoxide dismutase, lysozyme, antimicrobial peptides, and lectins, and the promotive effect of marine PNSB was stronger than that of freshwater PNSB. The cost of PNSB was calculated to be US$ 0.003–0.005 per kg feed; thus, marine PNSB is a cost-effective and feasible probiotic for shrimp aquaculture.

Shoc2 recognizes bacterial flagellin and mediates antibacterial Erk/Stat signaling in an invertebrate.

Flagellin is a key bacterial virulence factor that can stimulate molecular immune signaling in both animals and plants. The detailed mechanisms of recognizing flagellin and mounting an efficient immune response have been uncovered in vertebrates; however, whether invertebrates can discriminate flagellin remains largely unknown. In the present study, the homolog of human SHOC2 leucine rich repeat scaffold protein in kuruma shrimp (Marsupenaeus japonicus), designated MjShoc2, was found to interact with Vibrio anguillarum flagellin A (FlaA) using yeast two-hybrid and pull-down assays. MjShoc2 plays a role in antibacterial response by mediating the FlaA-induced expression of certain antibacterial effectors, including lectin and antimicrobial peptide. FlaA challenge, via MjShoc2, led to phosphorylation of extracellular regulated kinase (Erk), and the subsequent activation of signal transducer and activator of transcription (Stat), ultimately inducing the expression of effectors. Therefore, by establishing the FlaA/MjShoc2/Erk/Stat signaling axis, this study revealed a new antibacterial strategy in shrimp, and provides insights into the flagellin sensing mechanism in invertebrates.

Probiotic Effects of a Marine Purple Non-Sulfur Bacterium, Rhodovulum sulfidophilum KKMI01, on Kuruma Shrimp (Marsupenaeus japonicus).

Purple non-sulfur bacteria (PNSB) are used as probiotics in shrimp aquaculture; however, no studies have examined the probiotic effects of PNSB in shrimp at the gene expression level. In this study, we examined the effects of a marine PNSB, Rhodovulum sulfidophilum KKMI01, on the gene expression of kuruma shrimp (Marsupenaeus japonicus). Short-term (3 days) effects of R. sulfidophilum KKMI01 on the gene expression in shrimp were examined using small-scale laboratory aquaria experiments, while long-term (145 days) effects of R. sulfidophilum KKMI01 on the growth performance and gene expression were examined using 200-ton outdoor aquaria experiments. Gene expression levels were examined using qRT-PCR. Results of the short-term experiments showed the upregulation of several molting-related genes, including cuticle proteins, calcification proteins, and cuticle pigment protein, suggesting that PNSB stimulated the growth of shrimp. The upregulation of several immune genes, such as prophenoloxidase, antimicrobial peptides, and superoxide dismutase, was also observed. In the 145-day outdoor experiments, the average body weight at harvest time, survival rate, and feed conversion ratio were significantly improved in PNSB-treated shrimp, and upregulation of molting and immune-related genes were also observed. When PNSB cells were added to the rearing water, the effective dosage of PNSB was as low as 103 cfu/mL, which was more than a million times dilution of the original PNSB culture (2–3 × 109 cfu/mL), indicating that R. sulfidophilum KKMI01 provides a feasible and cost-effective application as a probiotic candidate in shrimp aquaculture.

Isobaric Tags for Relative and Absolute Quantitation-Based Proteomics Analysis Provides a New Perspective Into Unsynchronized Growth in Kuruma Shrimp (Marsupenaeus japonicus)

Unsynchronized growth is a common phenomenon in farmed crustaceans. The underlying molecular mechanism of unsynchronized growth of crustaceans is unclear. In this study, a comparative proteomic analysis focusing on growth differences was performed using kuruma shrimp Marsupenaeus japonicus, an economic crustacean species, as the model. The study analyzed kuruma shrimp at fast growth stage and steady growth stage from both fast growth group and slow growth group by an Isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic analysis method. A total of 1,720 proteins, including 12,291 peptides, were identified. Fifty-two and 70 differentially expressed proteins (DEPs) were identified in the fast growth stage and steady growth stage, respectively. Interestingly, 10 DEPs, including 14-3-3-epsilon-like, GPI, GPD1, MHC-1a, and MHC-1b, were presented in both growth stages. In addition, all these 10 DEPs shared the same expression tendency at these two growth stages. The results indicated that these 10 DEPs are potential growth biomarkers of M. japonicus. Proteins associated with faster growth of M. japonicus may promote cell growth and inhibit cell apoptosis through the Hippo signaling pathway. The fast growth group of M. japonicus may also achieve growth superiority by activating multiple related metabolic pathways, including glycolysis, glycerophospholipid metabolism and Citrate cycle. The present study provides a new perspective to explore the molecular mechanism of unsynchronized growth in crustacean species.

Carbon dioxide fluxes from mariculture ponds with swimming crabs and shrimps in eastern China: The effect of adding razor clams

Razor clams (Sinonovacula constricta) are one of the major polyculture species in mariculture ponds, along with swimming crabs (Portunus trituberculatus), in the coastal zones of China, and their effect on CO2 fluxes are poorly studied. CO2 fluxes from mariculture ponds with swimming crabs, kuruma shrimp (Marsupenaeus japonicus) (CS) and mariculture ponds with swimming crabs, kuruma shrimp and razor clams (with different clam densities from low to high, CSB1, CSB2, CSB3, respectively) were measured during the farming season. The CO2 fluxes of CS, CSB1, CSB2, and CSB3 were −25.37, −30.23, −28.81 and −12.18 mg m−2 h−1, respectively. All the ponds acted as CO2 sinks across the water-air interface. Our results showed that water pH was a good indicator of the CO2 flux, and a pH value of 8.24 was the critical value dividing the dataset into influxes or effluxes in the mariculture ponds. Net primary production was also significantly correlated with the CO2 flux and is considered a key driver regulating the CO2 flux. Our findings indicate that the low clam stocking density used in this study could enhance the CO2 sink across the water-air interface by promoting phytoplankton biomass and net primary production, whereas the CO2 sink may be weakened with an increase in clam density due to a decline in phytoplankton biomass and net primary production.

Antibacterial activity of four recombinant carbohydrate recognition domain proteins identified from the kuruma shrimp Marsupenaeus japonicus

The carbohydrate recognition domain (CRD) is the key component of C-type lectins (CTLs) with the capacity to recognize and eliminate invading pathogens. Herein, the recombinant proteins of four CRDs identified from the kuruma shrimp, Marsupenaeus japonicus, were produced and purified by an Escherichia coli expression system and affinity chromatography. Bacterial binding and antibacterial assays showed that the four CRDs displayed various bacterial binding and antibacterial activities against different bacteria. Among the four recombinant CRDs, His-CRD2–3 exhibited the broadest spectrum of bacterial binding and antibacterial activities against gram-negative bacteria (Vibrio parahaemolyticus, V. alginolyticus and V. harveyi) and gram-positive bacteria (Staphylococcus aureus and Micrococcus lysodeikticus). Moreover, the four recombinant CRDs showed different capacities to regulate the expression of several immune effector genes (MjCTL3, MjCTL4, MjCTL, Mjily and Mjsty), among which His-CRD2–3 displayed broader and stronger inductive effects on these immune effector genes. This study indicated that the four CRDs participated in immune defense by binding and killing bacteria and regulating the transcription of other immune effector genes. In addition, our results suggested that His-CRD2–3 might be a promising agent for the prevention and treatment of bacteriosis.

Interferon functional analog activates antiviral Jak/Stat signaling through integrin in an arthropod.

Drosophila Vago is a small antiviral peptide. Its ortholog in Culex mosquito was found to be an interferon-like cytokine that limits virus replication through activating Jak/Stat signaling. However, this activation is independent of Domeless, the sole homolog of vertebrate type I cytokine receptor. How Vago activates the Jak/Stat pathway remains unknown. Herein, we report this process is dependent on integrin in kuruma shrimp (Marsupenaeus japonicus). Shrimp Vago-like (MjVago-L) plays an antiviral role by activating the Jak/Stat pathway and inducing Stat-regulated Ficolin. Blocking integrin abrogates the role of MjVago-L. The interaction between MjVago-L and integrin β3 is confirmed. An Asp residue in MjVago-L is found criticalfor the interaction and MjVago-L's antiviral role. Moreover, Fak, a key adaptor of integrin signaling, mediatesMjVago-L-induced Jak/Stat activation. Therefore, this study reveals that integrin, as the receptor of MjVago-L, mediates Jak/Stat activation. The establishment of the MjVago-L/integrin/Fak/Jak/Stat/Ficolin axis provides insights into antiviral cytokine signaling in invertebrates.