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Interleukin 10 (IL-10), a pleiotropic cytokine, plays an essential role in multiple immunity responses. In the current study, the sequences of IL-10 family were identified from spotted knifejaw (Oplegnathus punctatus) whole genome, and O. punctatus IL-10 (OpIL-10) was cloned and characterized. OpIL-10 encodes 187 amino acids with a typical IL-10 family signature motif and predicted α-helices. It shared high identities with Notolabrus celidotus IL-10 and Epinephelus Lanceolatus IL-10. OpIL-10 was widely detected in healthy tissues, with the abundant expression in liver and skin. It was significantly up-regulated in the six immune-related tissues (liver, spleen, kidney, intestine, gill and skin) after infection against Vibrio harveyi and spotted knifejaw iridovirus (SKIV). Dual-luciferase analysis showed that OpIL-10 overexpression could suppress the activity of NF-κB. Meanwhile, OpIL-10 knockdown caused the down-regulation of five immune-related genes in JAK2/STAT3 signaling pathway and NF-κB signaling pathway, including IL-10R2 , TYK2 , STAT3 , NOD2 , and IκB. In addition, LPS and poly I:C stimulated expression of pro-inflammatory cytokines, including IL-6 , IL-1β , IL-8 , and IL-12 , were lower with recombinant OpIL-10 (rOp IL-10) than the control group, indicating the anti-inflammatory roles of rOpIL-10. Taken together, these results indicated OpIL-10 as a negative regulator in the inflammatory responses of spotted knifejaw against bacterial and viral infection, which would help us better understand the role of IL-10 in teleost immunity. • OpIL-10 was identified and characterized from spotted knifejaw (Oplegnathus punctatus). • OpIL-10 was expressed in multiple tissues and upregulated in six immune-related tissues after bacterial and viral infection. • OpIL-10 could regulate JAK2/STAT3 signaling pathway and NF-κB signaling pathway. • Recombinant OpIL-10 protein had an inhibitory effect on the expression of proinflammatory cytokines. • OpIL-10 plays a negatively regulator in the immune response against bacterial and viral infection in spotted knifejaw. [ABSTRACT FROM AUTHOR]

White spot syndrome virus (WSSV) is a fatal pathogen threatening global crustacean industry with no commercially available drugs to control. Herbal medicines have been widely used to treat a number of viral infections, which could offer a rich reserve for antiviral drug discovery. Here, we evaluated the inhibition activities of 30 herbal medicines against WSSV in Chinese mitten crab Eriocheir sinensis. A WSSV infection model in E. sinensis was firstly established in order to determine the antiviral effects of the plant extracts and to explore the potential action mechanisms. Results showed that the highest anti-WSSV activity was obtained by the treatment of Ophiopogon japonicus extract (93.03%, 100 mg/kg). O. japonicus treatment decreased viral loads in a dose-dependent manner and significantly improved the survival of WSSV-challenged crabs. O. japonicus reduced the expression of vital genes in viral life cycle in vivo , particularly for the immediate-early stage gene ie1. Further results indicated that O. japonicus could repress the JAK-STAT signaling pathway to block ie1 transcription. Moreover, O. japonicus could modulate certain immune genes such as the myosin , toll-like receptor , crustin , and prophenoloxidase in the interactions between WSSV and crabs. The up-regulated expression of pro-autophagic factors (Gabarap and Atg7) and elevated levels of antioxidant enzymes (SOD, CAT and GSH) suggested that O. japonicus may induce autophagy and attenuate WSSV-induced oxidative stress. Taken together, O. japonicus could inhibit WSSV proliferation and improve the survival of WSSV-challenged crabs. Thus, O. japonicus may have the potential to be developed as a preventive or therapeutic agent against WSSV, and its effective compounds merit further isolation and identification. • The WSSV infection model was established and evaluated in Eriocheir sinensis for the first time. • Ophiopogon japonicus restrains WSSV proliferation and improves survival of WSSV-challenged crabs. • O.japonicus represses the JAK/STAT signaling pathway to block the transcription of WSSV early gene. • Ophiopogon japonicus induces immune response in crabs. • O.japonicus induces apoptosis-related factors and enhances anti-oxidative activity in WSSV-challenged crabs. [ABSTRACT FROM AUTHOR]

Crustacean female sex hormone (CFSH) is responsible for sexual differentiation in crustaceans. The CFSH exhibited an interleukin-17 domain homologous to vertebrate IL-17, a family of inflammatory cytokines that play vital roles in immune defense. However, the immunoregulation of CFSH in crustaceans is a mystery. Therefore, this study aimed to investigate the immune regulatory roles of CFSH and CFSHR in the mud crab Scylla paramamosain. This study's immunofluorescence result revealed that Sp -CFSHR was highly expressed in granulocytes and semi-granulocytes but had moderate expression in hyalinocytes. The expression level of Sp-CFSH transcript in eyestalk ganglia and Sp - CFSHR in hemocytes were significantly up-regulated by the Poly (I:C) stimulation but significantly down-regulated in response to the lipopolysaccharide (LPS) stimulation. In our study, in vitro experiment exhibited that the nuclear transcription factors NF-κB signaling molecules (Sp-Dorsal and Sp-Relish), Sp-STAT , apoptosis-related gene Sp-IAP , and phagocytosis related gene (Sp-Rab5) expressions were significantly increased in hemocytes by recombinant CFSH (rCFSH) in vitro , but the pro-inflammatory cytokine gene (Sp-IL-16) expression was significantly suppressed. Finally, the rCFSH injection significantly up-regulated Sp-Dorsal , Sp-Relish , Sp-IAP , Sp-Caspase , Sp-ALF2 , and C-type lectin (Sp-CTL-B) expressions in hemocytes as well as enhanced the bacterial clearance of the mud crab. In conclusion, our results suggested that CFSH may be a counterpart of vertebrate IL-17 in crustaceans that can enhance innate immunity to defense against Vibrionaceae infection via the NF-κB and/or JAK-STAT signaling pathways. This study provides the first evidence that CFSH is involved in the immunoregulation in crustaceans and enriches the insight of neuroendocrine-immune regulatory system, which providing new ideas for disease prevention in the mud crab industry. • The Sp-CFSHR mRNA is highly expressed in semigranulocytes and granulocytes. • The Sp-CFSH and Sp-CFSHR mRNA respond to the challenge of Poly (I: C) and LPS. • rCFSH enhances phagocytosis of hemocytes and activates immune effector moleculesin mud crabs. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are non-coding RNA molecules that regulate gene expression in fish, but its regulatory mechanism of the skin mucosal immune response remains poorly understood. In order to investigate the immunological role of miRNAs, three sRNA libraries (mSC, mST1, mST2) from skin samples of crucian carp (Carassius auratus) infected with Aeromonas hydrophila at three time points (0, 6 and 12 hpi) were constructed and examined using Illumina Hiseq 2000 platform. All of the identified miRNA, rRNA and tRNA were 69444 (13.39%), 29550 (5.70%) and 10704 (2.06%) in skin, respectively. At 6 and 12 hpi, 829 and 856 miRNAs were differentially expressed, respectively. Among these DEMs, 53 known and 10 novel miRNAs were all significantly differentially expressed during early infection (p < 0.01). GO and KEGG enrichment analyses revealed that 118111 target-genes were primarily involved in cellular process, metabolic process, biological regulation and stress response, such as antigen processing and presentation, complement and coagulation cascades, phagosome, MAPK, TLR, NF-κB and JAK-STAT signaling pathways. These results will help to elucidate the mechanism of miRNAs involved in the skin mucosal immune response of crucian carp against Aeromonas hydrophila infection. • This was first report on skin miRNAs profiles of C. auratus upon A. hydrophila infection. • 829 and 856 DEMs in skin were identified at 6 and 12 hpi, respectively. • 10 novel miRNAs were significantly differentially expressed during early infection. • Antigen presentation, complement, MAPK, TLR and NF-κB pathways were significantly involved in skin mucosal immunity. [ABSTRACT FROM AUTHOR]

Lipopolysaccharide (LPS) is a classical pathogen-associated molecular pattern that can trigger strong inflammatory response mainly by TLR4-mediated signaling pathway in mammals, but the molecular mechanism of anti-LPS immunity is unclear in teleost fishes. In this study, we analyzed the gene expression features based on transcriptome analysis in Schizothorax prenanti (S. prenanti), after stimulation with two sources of LPS from Aeromonas hydrophila and Escherichia coli (Ah. LPS and Ecoli. LPS). 921 different expression genes (DEGs) after Ah. LPS stimulation and 975 DEGs after Ecoli. LPS stimulation were acquired, but only 706 and 750 DEGs were successfully annotated into the databases, respectively. Both of two groups of DGEs were significantly enriched into immune-related pathways by KEGG enrichment analysis, such as "Toll-like receptor signaling pathway", "Cytokine-cytokine receptor interaction" and "JAK-STAT signaling pathway". The annotated DEGs from Ah. LPS and Ecoli. LPS stimulation shared 470 DEGs, including 88 immune-related DEGs (IRGs) identified mainly by KEGG enrichment to immune-related signaling pathways. Among the shared IRGs, four pattern-recognition genes (TLR5 , TLR25 , PTX3 and C1q) were induced with high expression foldchange, and IFN-γ and relative genes also showed higher expression levels than control. Meanwhile, inflammatory signals were highlighted by upregulating the expression of inflammatory cytokines (IL-1β , IL-10 and IL-8). Moreover, some non-shared IRGs (including TLR2 and TLR4) were identified, suggesting that different sources of LPS own different potentials for the induction of immune gene expression. In conclusion, TLR5, TLR25, PTX3 and C1q may function as the sensing molecules to catch the invasion signal of LPS. The anti-LPS immune response may be involved into TLR25/TLR5-mediated inflammatory signals that regulate subsequently the activation of PTX3/C1q-modulated complement pathway upon the induction of PTX3 expression, and the crosstalk between IFN-γ and TLR signaling pathways in teleost fishes. This study will contribute to further explore the molecular mechanism of LPS-induced immunity in teleost fishes. • 921 different expression genes (DEGs) after Ah. LPS stimulation and 975 DEGs after Ecoli. LPS stimulation were acquired. • The stimulation with Ah. LPS and Ecoli. LPS shared 470 DEGs including 88 immune-related DEGs (IRGs). • TLR5, TLR25, PTX3 and C1q were predicted as the potential pattern-recognition molecules in LPS sensing. • IFN-γ-mediated signaling pathway may play an important role in protecting teleost fishes against LPS. [ABSTRACT FROM AUTHOR]

Streptococcus agalactiae is a major aquaculture pathogen infecting various saltwater and freshwater fish. To better understand the mechanism of the immune responses to S. agalactiae in wildtype zebrafish, the transcriptomic profiles of two organs containing mucosal-associated lymphoid tissues from S. agalactiae- infected and non-infected groups were obtained using RNA-seq techniques. In the intestines, 6735 and 12908 differently expressed genes (DEGs) were identified at 24 hpi and 48 hpi, respectively. Among 66 and 116 significantly enriched pathways, 15 and 21 pathways were involved in immune system or signal transduction at 24 hpi and 48 hpi, respectively. A number of genes involved in Toll-like receptor signaling pathway, RIG-I-like receptor signaling pathway, NOD-like receptor signaling pathway, T cell receptor signaling pathway, B cell receptor signaling pathway, Antigen processing and presentation, NF-kappa B signaling pathway and PI3K-Akt signaling pathway were significantly downregulated. In the skins, 3113 and 4467 DEGs were identified at 24 hpi and 48 hpi, respectively. Among 24 and 56 significantly enriched pathways, 4 and 13 pathways were involved in immune system or signal transduction at 24 hpi and 48 hpi, respectively. More immune-related signaling pathways including Leukocyte transendothelial migration, Cytokine-cytokine receptor interaction, PI3K-Akt signaling pathway, IL-17 signaling pathway, MAPK signaling pathway, TNF signaling pathway, Complement and coagulation cascades, Hematopoietic cell lineage and Jak-STAT signaling pathway were differently enriched for upregulated DEGs at 48 hpi, which were completely different from that in the intestines. Furthmore, comparative transcriptome analysis revealed that the downregulated 1618 genes and upregulated 1622 genes existed both at 24 hpi and 48 hpi for the intestine samples. In the skins, the downregulated 672 genes and upregulated 428 genes existed both at 24 hpi and 48 hpi. Three pathways related to immune processes were significantly enriched for downregulated DEGs both in the intestines and skins collected at 24 hpi and 48 hpi, which included Antigen processing and presentation, Intestinal immune network for IgA production and Hematopoietic cell lineage. Interaction network analysis of DEGs identified the main DEGs in the sub-network of complement and coagulation cascades both in the intestines and skins. Twenty of DEGs involved in complement and coagulation cascades were further validated by Real-time quantitative PCR. Altogether, the results obtained in this study will provide insight into the immune response of zebrafish against S. agalactiae XQ-1 infection in fatal conditions, and reveal the discrepant expression pattern of complement and coagulation cascades in the intestines and skins. • Intraperitoneal injections of S. agalactiae XQ-1 strain caused zebrafish mortality. • S. agalactiae XQ-1 infection suppressed the innate and adaptive immune responses of host in fatal conditions. • The expression changes in complement and coagulation cascades in intestine were different from that in skin. [ABSTRACT FROM AUTHOR]

Japanese pufferfish (Takifugu rubripes) is one of the main marine aquatic fish species cultured in Asia due to its high nutritional value. In recent years, disease caused by Vibrio harveyi infections have led to serious mortality in Japanese pufferfish industry. To understand the complex molecular mechanisms between V. harveyi and Japanese pufferfish, we performed a transcriptome analysis of liver and spleen samples from Japanese pufferfish at 1 and 2 day post-infection. Between-group comparisons revealed 922 genes that were significantly differentially expressed. The altered genes emphasized the function in several immune related pathways including MAPK signaling pathway, JAK-STAT signaling pathway, toll-like receptor signaling pathway, cytokine-cytokine receptor interaction and lysosomal pathway. The data generated in this study provided insight into the responses of Japanese pufferfish against V. harveyi at the transcriptome level, promoting our comprehensive understanding of immune responses for aquatic animal against V. harveyi. • Comparative transcriptome of Japanese pufferfish challenged with Vibrio harveyi were sequenced. • Between-group comparisons revealed 922 genes that were significantly differentially expressed. • Several immune related pathways were enriched. • Results will provide valuable resources for immune responses of aquatic animals against V. harveyi. [ABSTRACT FROM AUTHOR]

Abstract The Janus kinase/signal transducers and activators of transcription (JAK/STAT) signalling pathway plays a significant role in immune responses to pathogens. In invertebrates, three core components (Domeless, Hopscotch and STAT92E) of the JAK/STAT pathway were first identified in Drosophila melanogaster. In the present study, we report the cloning and characterisation of DOME, JAK and STAT from Eriocheir sinensis. Es DOME, Es JAK and Es STAT are 1299, 1110 and 794 amino acid proteins encoded by 4200, 3333 and 2385 bp open reading frames, respectively. Bioinformatics analysis revealed that all three share specific domains with corresponding functions. Quantitative real-time PCR showed that all three components were highly expressed in various tissues including the gill, hepatopancreas and hemocytes. Moreover, Es DOME, Es JAK and Es STAT were upregulated significantly in hemocytes after bacterial challenge. In addition, the JAK/STAT signalling pathway positively regulates antimicrobial peptide expression during the antibacterial immune response. Thus, the JAK/STAT pathway plays a critical role in bacterial immune responses in E. sinensis. Highlights • DOME, JAK and STAT from Eriocheir sinensis were cloned and characterised. • All three JAK/STAT pathway components have conserved domains and functions. • All three proteins are highly expressed in gill, hepatopancreas and hemocytes. • All three proteins are upregulated by bacterial challenge. • The JAK/STAT pathway stimulates antimicrobial peptide expression in E. sinensis. [ABSTRACT FROM AUTHOR]

Dissolved oxygen (DO) is essential for teleosts, and fluctuating environmental factors can result in hypoxic stress in the golden pompano (Trachinotus blochii). However, it is unknown whether different recovery speeds of DO concentration after hypoxia induce stress in T. blochii. In this study, T. blochii was subjected to hypoxic conditions (1.9 ± 0.2 mg/L) for 12 h followed by 12 h of reoxygenation at two different speeds (30 mg/L per hour and 1.7 mg/L per hour increasing). The gradual reoxygenation group (GRG), experienced DO recovery (1.9 ± 0.2 to 6.8 ± 0.2 mg/L) within 3 h, and the rapid reoxygenation group (RRG), experienced DO recovery (1.9 ± 0.2 to 6.8 ± 0.2 mg/L) within 10 min. Physiological and biochemical parameters of metabolism (glucose, glycegon, lactic acid (LD), lactate dehydrogenase (LDH), pyruvic acid (PA), phosphofructokinase (PFKA), and hexokinase (HK), triglyceride (TG), lipoprotein lipase (LPL), carnitine palmitoyltransferase 1 (CPT-1)) and transcriptome sequencing (RNA-seq of liver) were monitored to identify the effects of the two reoxygenation speeds. Increased LD content and increased activity of LDH, PA, PFKA, and HK suggested enhanced anaerobic glycolysis under hypoxic stress. LD and LDH levels remained significantly elevated during reoxygenation, indicating that the effects of hypoxia were not immediately alleviated during reoxygenation. The expressions of PGM2 , PFKA , GAPDH , and PK were increased in the RRG, which suggests that glycolysis was enhanced. The same pattern was not observed in the GRG. Additionally, In the RRG, reoxygenation may promote glycolysis to guarantee energy supply. However, the GRG may through the lipid metabolism such as steroid biosynthesis at the later stage of reoxygenation. In the aspect of apoptosis, differentially expressed genes (DEGs) in the RRG were enriched in the p53 signaling pathway, which promoted cell apoptosis, while DEGs in the GRG seem to activate cell apoptosis at early stage of reoxygenation but was restrained latterly. DEGs in both the RRG and the GRG were enriched in the NF-kappa B and JAK-STAT signaling pathways, the RRG may induce cell survival by regulating the expression of IL-12B , COX2 , and Bcl-XL , while in the GRG it may induce by regulating the expression of IL-8. Moreover, DEGs in the RRG were also enriched in the Toll-like receptor signaling pathway. This research revealed that at different velocity of reoxygenation after hypoxic stress, T. blochii would represent different metabolic, apoptotic and immune strategies, and this conclusion would provide new insight into the response to hypoxia and reoxygenation in teleosts. [Display omitted] • Glucose and lipid levels returned to normal during reoxygenation after hypoxia. • In the GRG, the expression of FDFT1 and HSD17B7 may guarantee the lipid metabolism. • In the RRG, the expression of Fas , CASP8 and CASP10 may promote apoptosis. • In the RRG, the expression of IL-12B , COX2 , and Bcl-XL may activate cell survival. [ABSTRACT FROM AUTHOR]

Hirame novirhabdovirus (HIRRV) is a fish rhabdovirus belonging to family Rhabdoviridae, genus Novirhabdovirus , which is highly contagious and virulent, and causes hemorrhagic disease in many fish species. In the present work, the whole genome sequence of HIRRV strain CNPo2015 that previously isolated from cultured flounders was obtained using high-throughput sequencing. It consists of 10,998 nucleotides and encodes six viral proteins arranged in order of 3′-N-P-M-G-NV-L-5′. Among Novirhabdovirus , L protein of CNPo2015 possessed the lowest amino acid sequence divergence with HIRRV isolate CA 9703 and HIRRV 080113, and the highest with Snakehead rhabdovirus. Furthermore, the immune response of Hirame natural embryo (HINAE) cell line to HIRRV infection was characterized by RNA-seq, and the results showed that 1976 differentially expressed genes (DEGs) including 1219 up-regulated and 727 down-regulated genes were identified in the HINAE cells infected with HIRRV at 48 h post infection (hpi). Several KEGG pathways were significantly enriched in the viral infected cells, such as cytokine-cytokine receptor interaction, JAK-STAT signaling pathway, cell cycle, apoptosis, RIG-I-like receptors signaling pathway and P13K-AKT signaling pathway. Post viral infection, the flow cytometric Annexin V/PI assay found that apoptotic rate of HINAE cells showed a slight increase within 3 days and then the early and late apoptotic rate were significantly increased to 41 ± 2.65% and 12.37 ± 2.61% at day 4, respectively. Meanwhile, qRT-PCR results also showed that six apoptosis-related genes (BCL2L1, CASPASE 3, CASPASE 10, FAS, AKT and CDK1) were significantly upregulated. This investigation has not only enriched our knowledge of sequence difference characteristics between CNPo2015 and other Novirhabdoviruses , but also provided a data basis for deeper understanding of immune responses in flounder cells post viral infection. • Genome sequence of HIRRV strain CNPo2015 was obtained and compared with other Novirhabdoviruses. • The immune response of HINAE cells to HIRRV infection was systematically characterized by RNA-seq. • HIRRV infection upregulated apoptotic genes and induced cellular apoptosis of HINAE cells. [ABSTRACT FROM AUTHOR]