Fish and shellfish immunology reports最新文献

Aquaculture plays an important role in contributing to global food security and nutrition; thus, the intensification and diversification of aquaculture are increasingly considered. However, paralleling the development of the industrial scale in aquaculture, the occurrence of diseases is always an important issue that causes great losses in economics. The finding of approaches that not only improve culture production but also reduce the impact of diseases in cultured animals is crucially essential. Previously, several studies have addressed the potential application of feed additives, such as prebiotics, probiotics, synbiotics, and microbial-derived metabolites (including short-chain fatty acids-SCFAs), in aquaculture. In this review, we provide an update focusing on the health benefits of dietary supplementation with a type of SCFAs, butyrate, and its producer, Clostridium butyricum, including their effects on growth, feed utilization, body composition, intestinal structure and function, antioxidant activity, immune response, and tolerance against stress and infection in aquatic animals. The outcomes of this study may indicate more benefits of the use of C. butyricum than that of butyrate (and its forms). This review provides general knowledge of the efficacy of butyrate and C. butyricum in aquaculture.

The development and validation of the recombinant 9E1 monoclonal antibody against channel catfish IgM is described. The variable heavy and light chain domains of the 9E1 hybridoma were cloned into murine IgG1 and IgK expression vectors. These expression plasmids were co-transfected into 293F cells and mature IgG was purified from culture supernatant. It is demonstrated that the recombinant 9E1 monoclonal antibody binds to soluble IgM in ELISA and ELISPOT assays and to membrane-bound IgM by immunofluorescence with different B-cell types. The recombinant 9E1 monoclonal antibody will be a valuable tool in the continued examination of the channel catfish adaptive immune system

As a pleiotropic cytokine in the interleukin (IL)-12 family, IL-27β plays a significant role in regulating immune cell responses, eliminating invading pathogens, and maintaining immune homeostasis. Although non-mammalian IL-27β homologs have been identified, the mechanism of whether and how it is involved in adaptive immunity in early vertebrates remains unclear. In this study, we identified an evolutionarily conserved IL-27β (defined as OnIL-27β) from Nile tilapia (Oreochromis niloticus), and explored its conserved status through gene collinearity, gene structure, functional domain, tertiary structure, multiple sequence alignment, and phylogeny analysis. IL-27β was widely expressed in the immune-related tissues/organ of tilapia. The expression of OnIL-27β in spleen lymphocytes increased significantly at the adaptive immune phase after Edwardsiella piscicida infection. OnIL-27β can bind to precursor cells, T cells, and other lymphocytes to varying degrees. Additionally, IL-27β may be involved in lymphocyte-mediated immune responses through activation of Erk and JNK pathways. More importantly, we found that IL-27β enhanced the mRNA expression of the Th1 cell-associated cytokine IFN-γ and the transcription factor T-bet. This potential enhancement of the Th1 response may be attributed to the activation of the JAK1/STAT1/T-bet axis by IL-27β, as it induced increased transcript levels of JAK1, STAT1 but not TYK2 and STAT4. This study provides a new perspective for understanding the origin, evolution and function of the adaptive immune system in teleost.

Spleen tyrosine kinase (Syk) is reported to be involved in activating the autophagy. Recently, a homologue of Syk was identified from Pacific oyster Crassostrea gigas (defined as CgSyk). In the present study, the molecular characteristics of CgSyk and its regulation mechanism in autophagy were investigated in oyster C. gigas. The full-length cDNA of CgSyk was of 4566 bp with an open reading frame (ORF) of 1989 bp. CgSyk encoded a polypeptide of 662 amino acids, containing two Src homology 2 (SH2) domains and one tyrosine kinase catalytic (TyrKc) domain. The deduced amino acid sequence of CgSyk shared low similarity with the previously identified Syks from other species. In the phylogenetic tree, CgSyk was first clustered with Crassostrea virginica CvSyk, and then classified into a branch of invertebrate Syks. In CgSyk-RNAi oysters, the mRNA expressions of CgLC3, CgP62, CgBeclin-1 and CgATG5 in haemocytes decreased significantly at 12 h after Vibrio splendidus stimulation. At the same time, the abundance of CgLC3Ⅱ in haemocytes, and the autophagy rate of haemocytes in CgSyk-RNAi oysters decreased significantly at 12 h after V. splendidus stimulation. All the results collectively suggested that CgSyk regulated the autophagy through inducing the mRNA expressions of autophagy-related genes and the cleavage of CgLC3 to defend against bacterial invasion in oysters.

This short paper on yellow head virus Type-1 (YHV-1) of shrimp describes preliminary research on the potential for using YHV-1 attenuated in insect cells to protect shrimp against yellow head disease (YHD). YHV-1 can cause severe mortality in the cultivated shrimp Penaeus (Penaeus) monodon and Penaeus (Litopenaeus) vannamei.  No practical vaccination has been reported. The C6/36 mosquito cell cultures inoculated with YHV-1 become positive by PCR and by immunocytochemistry (immunopositive) for up to 30 split-cell passages. Shrimp injected with homogenates from low-passage cultures die from typical YHV-1 disease while shrimp injected with homogenates from high passage cultures do not, even though they become PCR positive and immunopositive for YHV-1. This suggested that viral attenuation had occurred during insect-cell passaging, and it opened the possibility of using homogenates from high-passage insect cultures as a vaccine against YHV-1. To test this hypothesis, homogenates from 30th-passage, YHV-positive cultures were injected into shrimp followed by challenge with virulent YHV-1. Controls were injected with homogenate from 30th-passage, naive (normal stock) insect-cell cultures. No shrimp mortality occurred following injection of either homogenate, but shrimp injected with the YHV-1 homogenate became both RT-PCR positive and immunopositive. Upon challenge 10 days later with YHV-1, mortality in shrimp injected with naive insect-cell homogenate was 100% within 7 days post-challenge while 100% mortality in the YHV-1 homogenate group did not occur until day 9 post-challenge. Kaplan-Meier log-rank survival analysis revealed that survival curves for the two groups were significantly different (p < 0.001). The cause of delay in mortality may be worthy of further investigation.

Polyinosinic-polycytidylic acid (poly I:C) is a synthesized analogue of viral double-strand RNA and considered as a potential immunostimulant in aquaculture. MicroRNAs (miRNAs) have been reported to play important roles in the development of the immune system and in regulation of host antiviral responses. In our earlier study, it was found that poly I:C pre-treatment could stimulate the resistance against cyprinid herpesvirus 2 (CyHV-2) infection and enhance the antiviral immune response in gibel carp. To understand the role of miRNAs in regulating the host response to poly I:C treatment, we investigated the expression profiles of miRNAs in the head kidney of poly I:C-treated gibel carp with small RNA sequencing technology. When compared with the untreated group, a total of 24 differentially expressed miRNAs were identified in the poly I:C-stimulated fish, among which, 7 and 17 miRNAs were upregulated and downregulated, respectively. Analysis of target genes of these differentially expressed miRNAs found that most targeted mRNAs were involved in catalytic activity, peptidase activity and endopeptidase activity, and were enriched in the metabolic, protein processing in endoplasmic reticulum and oxidative phosphorylation signaling pathways, suggesting that poly I:C could alter the expression of metabolism-related miRNAs in the kidney of gibel carp. Besides, it was noted that some immune-related miRNAs, including inflammation-related miRNAs (miR-192 and miR-731) and interferon-related miRNAs (miR-194a and miR-122), were downregulated after poly I:C treatment. In summary, it was found that poly I:C could regulate the cellular levels of specific miRNAs involved in metabolism and immune responses in the head kidney of gibel carp, which may increase the capacity of the immune cells to fight against pathogens infection.

Peroxiredoxins (Prxs) widely exist in organisms and can prevent oxidative damage. Here, the characterization and biological function of NdPrx3 from Neocaridina denticulata sinensis were analyzed. The coding sequence of NdPrx3 consists of 684 bp open reading frame (ORF), encoding 227 amino acids with a predicted molecular weight of 24.7 kDa and theoretical pI 6.49. Multiple sequence alignments showed that the conserved domains of NdPrx3, including catalytic triad, dimer interface, decamer interface, peroxidatic, and resolving cysteines, were similar to those of other organisms. The phylogenetic relationship demonstrated that NdPrx3 clustered in the Prx3 class. The highest relative expression of NdPrx3 mRNA was confirmed in gill among the nine tissues from healthy shrimp. The transcript level of NdPrx3 was significantly upregulated from 0 h to 48 h and decreased in 72 h under copper challenge, indicating that NdPrx3 may play an important role in the copper challenge of N. denticulata sinensis. In addition, NdPrx3 was recombinantly expressed in E. coli and purified to one band on SDS-PAGE. The DNA protection of rNdPrx3 was verified. The enzymatic assay of the recombinant NdPrx3 indicated that it had the oxidoreductase function and was stable at a low temperature (10–30 °C).

Salmonid novirhabdovirus (IHNV) causes infectious haematopoietic necrosis (IHN) in salmonid species. Despite an injectable plasmid-based DNA vaccine of the glycoprotein (G) gene is effective, there are no oral vaccines for mass vaccination of rainbow trout (Oncorhynchus mykiss) fry. Recombinant baculoviruses were generated, used in cabbage looper (Trichoplusia ni) insect larvae to produce IHNV G and IHNV G-C5a proteins. Western blotting and chemiluminescence assays confirmed the expression of recombinant proteins, which were added to the fish feeding and top-coated with unflavored gelatin binder. Commercial rainbow trout were fed with experimental diets containing either IHNV G or IHNV G-C5a proteins for 2 weeks, and boosted 4 weeks after. Four weeks post-booster, fish were challenged with IHNV by immersion. Survival upon the infection challenge was evaluated. Spleen were sampled at 7 and 14 days post infection (dpi). Non-vaccinated and IHNV G fed trout reached a mortality of 91.7 and 97.6%, and 70.9 and 88.4%, respectively at 8 and 15 dpi. The IHNV G-C5a fed group exhibited a reduced mortality of 51.2% at 8 dpi, reaching 81.7% at 15 dpi, suggesting some level of antiviral protection. The individual viral load was measured by RT-qPCR detection of IHNV N gene, showing no significant difference across experimental groups. The transcription modulation of selected immune response markers was evaluated across experimental groups, including Type I IFN-a, Mx-1, CD4, and IgM. Further study is needed to assess how new oral vaccines may become effective to mitigate IHNV pathogenesis in juvenile trout by modulating the host immune response to protect towards IHNV exposure.

The current study aimed at assessing the immunostimulatory properties of silver nanoparticles (AgNPs) on Labeo rohita, and understanding how it affects the growth, cellular ultrastructure, the expression level of immune genes, and infection risk from Aeromonas hydrophila. Fish (avg wt: 30.1±3.26 g) were fed diets with four separate AgNP inclusion levels (0 µgKg⁻¹ [basic diet, T1], 10 µgKg⁻¹ [T2], 15 µgKg⁻¹ [T3], and 20 µgKg⁻¹ [T4]) for 56 days. After the feeding trial, growth, histological, immunological parameters, and protective immune response against A. hydrophila were assessed. The fish in the treatment groups including T1(control), the T3 growth indices, such as specific growth rate (7.56±0.26) and percent weight gain (231.05±3.21), was statistically higher (P < 0.05). In the immunological and oxidative parameters, levels of SOD and catalase decreased in correlation with a rise in the inclusion doses of AgNP in the liver, and a reduction in catalase values was recorded in the gill. With the addition of AgNP, the NBT value was decreased in the gill, and T3 had a considerably larger (P<0.05) value in the liver (0.493±0.02). The kidney of the L. rohita fed AgNP (0 and20 µgKg⁻¹ AgNP) showed expansion through Bowman's gaps, severing of glomeruli with haemorrhage, as well as atrophic spots between its gaps. The liver showed fibrosis, karyolysis, and the removal of the hepatocytes wall. The gill, liver, kidney, and muscle of fish-fed diets supplemented with AgNP, showed that interleukin-8 (IL-8), and cyclooxygenase-2 (COX-2), were up-regulated. Expression was considerably higher in T3 compared with the control. However, the control group that wasn't given AgNP supplemented diet had increased levels of TGF-beta. Additionally, fish on the T3 diet showed much greater post-challenge survival rates (90%). These findings strongly suggest that dietary inclusion of AgNP (at 10 and 15 µgKg⁻¹ feed) enhances growth, health, and protective immune response against A. hydrophila.

The polian vesicle and coelom of sea cucumber Apostichopus japonicus were full of coelomic fluid in which many types of coelomocytes with different functions were suspended. Our previous work has indicated the differences of coelomocytes between two sites mainly in subtype proportion, non-specific immune enzymes activities and several immune-related genes expression levels in healthy A. japonicus. However, the functional similarities and differences of coelomic fluid in two sites including the coelom and polian vesicle after pathogenic infection still remain unclear. Here, we investigated the changes of the total coelomocyte density (TCD) and differential coelomocyte density (DCD) after pathogen infection by Vibrio splendidus in coelom and polian vesicle. After infected by V. splendidus, the TCD in the coelom and polian vesicle rapidly declined at 12 h, and then the TCD in the coelom showed a stably ascending trend, while the TCD in the polian vesicle reached a peak at 24 h post infection (hpi), and then showed a continuously decline trend from 24 hpi to 72 hpi followed by a slow elevation until recovering the normal level from 72 hpi to 96 hpi. Then the activities of acidic phosphatase (ACP), alkaline phosphatase (AKP), catalase (CAT) and superoxide dismutase (SOD) were determined to evaluate the response of cell-free coelomic fluid to V. splendidus infection. The activities of ACP, AKP and CAT showed similar trends in the coelom and polian vesicle. The SOD activity significantly increased in the polian vesicle, whereas it exhibited a decreasing trend in the coelom. Finally, the expression profiles of nine immune-related genes including Aj-MyD88, Aj-IRAK4, Aj-i-Lys, Aj-Rel, Aj-p50, Aj-DMBT1, Aj-CDC, Aj-Rrp15 and Aj-Fibrinogen C were detected after V. splendidus challenge. The results suggested all the detected genes were significantly up-regulated both in the coelom and polian vesicle, and the expression levels of these genes in two sites shared similar trends except Aj-MyD88 and Aj-DMBT1. This research provides a new insight into the differentially immune roles of coelomic fluid and coelomocytes in polian vesicle and coelom response to bacterial infections and supplements comprehensive resources for better understanding the innate immune response of A. japonicus.