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Fish & Shellfish Immunology
Academic Press
Fish & Shellfish Immunology

Academic Press

1050-4648

Fish & Shellfish Immunology/Journal Fish & Shellfish ImmunologySCIISTP
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    Molecular cloning, expression analysis of interleukin 17D (cysteine knot cytokine) from Amphiprion clarkii and their functional characterization and NF kappa B pathway activation using FHM cells

    Liyanage, D. S.Omeka, W. K. M.Nadarajapillai, KishanthiniLim, Chaehyeon...
    10页
    查看更多>>摘要：Interleukin 17D (IL-17D), a pro-inflammatory cytokine, is a signature cytokine of T helper 17 (Th17) cells. However, studies characterizing the functions of IL-17D in teleost are scarce. Therefore, we aimed to characterize the properties of IL-17D in Amphiprion clarkii. We performed spatial and temporal expression, AcIL-17D-mediated antibacterial and inflammatory gene expression, NF kappa B pathway-related gene expression analyses, and bacterial colony counting and cell protection assays. We found that AcIL-17D contains a 630 bp coding sequence and encodes 210 amino acids. The spatial expression analysis of AcIL-17D in 12 tissues showed ubiquitous expression, with the highest expression in the brain, followed by blood and skin. Temporal expression analysis of AcIL-17D in blood showed upregulated expression at 6 and 24 h (polyinosinic: polycytidylic acid and lipopolysaccharide), 12 h (all stimulants), and 48 h (polyinosinic: polycytidylic acid and Vibrio harveyi). AcIL-17D expression in the blood gradually decreased at later hours in response to all the stimulants. After treatment of fathead minnow (FHM) cells with different recombinant AcIL-17D concentrations, the downstream gene expression analysis showed increased expression of antimicrobial genes in the FHM cells, namely [NK-Lysin (NKL), Hepcidin antimicrobial peptide-1 (HAMP-1), Defensin-beta (DEFB1)] and some inflammatory genes such as IL-1 beta, TNF-alpha, IL-11, and STAT3. Further nuclear factor kappa B (NF kappa B) subunits (NF kappa B1, NF kappa B2, RelA, and Rel-B) showed upregulated gene expression at 12 and 24 h. The bacterial colony counting assay using FHM cells showed lower bacterial colony counts in rAcIL-17D-treated cells than in control. Furthermore, the Water-Soluble Tetrazolium Salt (WST -1) assay confirmed the ability of rAcIL-17D in the protection of FHM cells from bacterial infection and conducted the Hoechst 33342 staining upon treatment with rAcIL-17D and rMBP. Therefore, our findings provide important insights into the activation of IL-17D pathway genes in FHM cells, the protective role of AcIL-17D against bacterial infection, and host defense mechanisms in teleost.
      原文链接:
    • NSTL
    • Elsevier

    Effect of low-frequency noise on the survival rate and immunity of infected Vibrio parahaemolyticus sea slug (Onchidium reevesii)

    Tu, ZhihanTang, LiusiqiaoYang, HangZhang, Xiaoming...
    10页
    查看更多>>摘要：Anthropogenic noise in the marine environment has become a global environmental pollutant that affects the behavior, physiology and immunity of marine animals. However, the resistance of marine animals to pathogens while under the influence of noise is a topic that has received little attention. To assess the immune defense response of sea slugs against pathogens when exposed to low frequency noise, we performed 120 h exposure experiments on sea slugs after a Vibrio parahaemolyticus application in low frequency noise at 500 Hz and 1000 Hz. We found that after the infection with V. parahaemolyticus, the survival rate of the sea slugs decreased, the apoptosis rate and reactive oxygen species (ROS) production of hemocytes increased significantly (P < 0.05), the proliferation of hemocytes accelerated, the activities of enzymes such as superoxide dismutase (SOD), catalase (CAT), alkaline phosphatase (AKP), alanine transaminase (ALT) and lysozyme (LZM) in the hepatopancreas increased significantly, and the expression of TNF signaling pathway-related genes (TNF-a, FADD, Caspase 8, Caspase 3) and Hsp70 genes were generally upregulated. In addition, exposure of sea slug after infected with V. parahaemolyticus to low frequency noise resulted in a significant increase in both antioxidant and immune parameters, which were positively correlated with frequency. The results showed that noise frequency and exposure time had an interactive effect on the above indicators. In summary, low-frequency noise exposure increases the risk of pathogenic infections in sea slugs and exacerbates the negative effects on the antioxidant capacity and immune metabolism of the organism.
      原文链接:
    • NSTL
    • Elsevier

    Rice protein concentrate as a fish meal substitute in Oreochromis niloticus: Effects on immune response, intestinal cytokines, Aeromonas veronii resistance, and gut microbiota composition

    Basili, MarcoAarestrup, Frank M.Pansera, LidiaMirto, Simone...
    14页
    查看更多>>摘要：The potential of rice protein concentrate (RPC) to substitute fishmeal (FM) protein in the diet of Oreochromis niloticus was assessed in a five-month-long feeding trial. Fishmeal protein was replaced by RPC at rates of 0% (control), 25%, 50%, and 75% (RPC0, RPC25, RPC50, and RPC75, respectively). RPC25 had no significant effect on antioxidant capacity (total antioxidant capacity; superoxide dismutase, catalase, and glutathione peroxidase activities) and immune indices (lysozyme, nitric oxide, antiprotease, and bactericidal activities) after one, two, and five months of feeding, while the values for these parameters were significantly lower in the RPC75 group compared to those in the RPC0 group. The RPC25 group showed higher mRNA levels of the intestinal cytokines IL-1 beta, IL-10 beta, TGF-beta, and TNF-alpha than the control group. In fish affected by Aeromonas veronii, the highest significant cumulative mortality was recorded in the RPC75 group, followed by the RPC50, RPC25, and control groups. Gut microbiome analyses showed a reduction in microbial diversity in response to the addition of RPC, regardless of the RPC content, and the composition of the community of the RPC samples differed from that of the control. RPC-enriched diets resulted in higher relative abundances of Bacteroidetes and Fusobacteria in the gut compared to that in the gut of the control fish. In summary, RPC can be used to replace up to 25% of the FM protein in the diet of O. niloticus, while improving the antioxidant capacity, immunocompetence, and disease resistance of the fish.
      原文链接:
    • NSTL
    • Elsevier

    Molecular characterization and expression analysis of septin gene family and phagocytic function of recombinant septin 2, 3 and 8 of starry flounder (Platichthys stellatus)

    Sohn, Min-YoungChoi, Kwang-MinJoo, Min-SooKang, Gyoungsik...
    12页
    查看更多>>摘要：Septin is an evolutionarily conserved family of GTP-binding proteins. Septins are known to be involved in a variety of cellular processes, including cell division, chromosome separation, cell polarity, motility, membrane dynamics, exocytosis, apoptosis, phagocytosis, DNA damage responses, and other immune responses. In this study, the sequences of the septin gene family of starry flounder were obtained using NGS sequencing, and the integrity of the sequences was verified through cloning and sequencing. At first, the amino acid sequence was annotated using the cDNA sequence, and then, the gene sequence was verified through multiple sequence alignment and phylogenetic analyses using the related conserved sequences. The septin gene family was classified into three subgroups based on the phylogenetic analysis. High conservation within the domain and homology between the genes reported in different species were confirmed. The expression level of septin gene family mRNA in each tissue of healthy starry flounder was evaluated to confirm the tissue- and gene-specific expression levels. Additionally, as a result of the analysis of mRNA expression after simulated pathogen infection, significant expression changes and characteristics were confirmed upon infection with bacteria (Streptococcus parauberis PH0710) and virus (VHSV). Based on the current results and that of previous studies, to confirm the immunological function, Septin 2, 3, and 8 were produced as recombinant proteins based on the amino acid sequences, and their role in phagocytosis was further investigated. The results of this study indicate that septin gene family plays a complex and crucial role in the host immune response to pathogens of starry flounder.
      原文链接:
    • NSTL
    • Elsevier

    The long noncoding RNA LTCONS5539 up-regulates the TRAF6-mediated immune responses in miiuy croaker (Miichthys miiuy)

    Pan, JiajiaZheng, WeiweiSun, YuenaXu, Tianjun...
    8页
    查看更多>>摘要：With the further study of long noncoding RNAs (lncRNAs), an increasing number of biological studies have demonstrated that lncRNAs are involved in various physiological processes, including cell proliferation, apoptosis, invasion, development and disease states. However, unlike mammals, little is known about the role of lncRNAs in the innate immunity of teleost fish. Here, we identify a lncRNA, named LTCONS5539, as critical role in the antiviral and antibacterial response of miiuy croaker and the results showed that lncRNA LTCONS5539 plays a critical regulatory role on TRAF6. Firstly, we found that LPS and poly(I:C) can up-regulate the expression of lncRNA LTCONS5539. Elevated lncRNA LTCONS5539 is capable of increasing the production of inflammatory factors and antiviral genes. Furthermore, the over-expression of lncRNA LTCONS5539 increases the expression of TRAF6 which was confirmed by qPCR and western blotting. On these foundations, we also proved that lncRNA LTCONS5539 modulates innate immunity through TRAF6-mediated immune responses through dual luciferase reporter assay. These results will help to further understand the immunomodulatory mechanisms of lncRNA in teleost fish.
      原文链接:
    • NSTL
    • Elsevier

    Litopenaeus vannamei peritrophin interacts with WSSV and AHPND-causing V. parahaemolyticus to regulate disease pathogenesis

    Chen, Yi-LunKumar, RamyaLiu, Chun-HungWang, Han-Ching...
    12页
    查看更多>>摘要：Peritrophins are peritrophic membrane (PM) proteins that can interact with chitin fibers via chitin-binding domains. Peritrophins have essential roles in providing porosity and strength to the PM that lines the shrimp midgut. Acute hepatopancreatic necrosis disease (AHPND), caused by strains of V. parahaemolyticus, is known to initially colonize the shrimp stomach and simultaneously disrupt its structural barriers (e.g., cuticle or epithelial tissues) to reach the hepatopancreas. Although stomach and hepatopancreas were identified as target tissues involved in AHPND pathogenesis, our results indicated that peritrophin in peritrophic membrane has a crucial role in determining not only colonization of AHPND-causing bacteria but also their tissue distribution. As the interaction between LvPeritrophin (LvPT) and WSSV (white spot syndrome virus) is not well understood, we noted that LvPT expression was upregulated in shrimp stomach challenged with either WSSV or AHPND. In an in vitro pathogen binding assay, there was strong binding of recombinant LvPT WSSV and AHPND-causing V. parahaemolyticus, and various bacteria. Furthermore, dsRNA-mediated LvPT silencing inhibited WSSV gene expression and viral genome replication. However, downregulation of LvPT gene expression increased copies of AHPND-causing bacteria in shrimp digestive tract, and facilitated bacterial colonization in stomach. In conclusion, we speculated that LvPT might regulate bacterial colonization during AHPND, whereas in WSSV infection, LvPT silencing favored the host. Although recombinant LvPT had strong binding with WSSV, the precise role of LvPT in WSSV infection needs further investigation. These findings increased our understanding of host-pathogen interactions in AHPND and WSSV infection that can be applied in shrimp aquaculture for developing effective antibacterial and antiviral strategies.
      原文链接:
    • NSTL
    • Elsevier

    Effects of the Clostridium butyricum on growth performance, antioxidant capacity, immunity and disease resistance of Litopenaeus Vannamei fed with cottonseed protein concentrate (CPC) replacement of fishmeal in diet

    Li, WeikangPan, LingLiu, HongyuTan, Beiping...
    9页
    查看更多>>摘要：Clostridium butyricum (CB) is a gram-positive bacterium that secretes short-chain fatty acids such as butyric acid and so on. An 8-week feeding trial was conducted to investigate the effects of CB on the growth performance, antioxidant capacity, immunity and resistance to Vibrio parahaemolyticus in Litopenaeus Vannamei fed with cottonseed protein concentrate (CPC) replacement of fishmeal. Six iso-nitrogenous (40%) and iso-lipidic (6%) diets were formulated including a positive control group (PC, 25% fishmeal), a negative control group (NC, CPC replaced 30% of fishmeal protein), and 0.03% (C1, 3 x 10(8) CFU/kg), 0.12% (C2, 1.2 x 10(9) CFU/kg), 0.48% (C3, 4.8 x 10(9) CFU/kg) and 1.92% (C4, 1.92 x 10(10) CFU/kg) CB were supplemented on the negative control group (NC). After the feeding trial, the remaining shrimp in each treatment group were subjected to a challenge experiment with Vibrio parahaemolyticus. The results indicated that weight gain rate (WGR), specific growth rate (SGR) in C4 group were significantly lower than those in PC and C2 groups (P < 0.05); the feed conversion ratio (FCR) was significantly higher than that of PC and C2 groups (P < 0.05). There was no significant difference in survival rate (SR) among all groups (P > 0.05). Compared to the PC and NC groups, the total superoxide capacity, superoxide dismutase and lysozyme were significantly higher in the C4 group (P < 0.05); the glutathione peroxidase, acid phosphatase and alkaline phosphatase were significantly higher in the C3 group (P < 0.05); and the malondialdehyde was significantly lower in the C4 group (P < 0.05). The relative mRNA expressions of Toll receptor (TLR), innate immune deficiency gene (IMD), penaiedin3a (Pen3) were significantly down-regulated in the NC group than those in the PC group (P < 0.05). In addition, the relative mRNA expressions of TLR, IMD and Pen3 were significantly up-regulated in all groups supplemented with CB than those in the NC group (P < 0.05). Moreover, the cumulative mortality rate in the NC group was not significantly different from the PC group (P > 0.05) and was significantly higher than those in the C3 and C4 groups (P < 0.05). In conclusion, the CB supplementation on the basis of CPC replacement of 30% fishmeal protein enhanced significantly the antioxidant capacity, immunity and disease resistance of shrimp and improved its growth performance. Therefore, considering the factors of the growth, immunity and disease resistance, the CB supplementation of 0.12%-0.48% (1.2 x 10(9) CFU/kg-4.8 x 10(9) CFU/kg) was recommended in the diet of L. vannamei based on the results of this experiment.
      原文链接:
    • NSTL
    • Elsevier

    Protective effects of tree peony seed protein hydrolysate on Cd-induced oxidative damage, inflammation and apoptosis in zebrafish embryos

    Li, YanWang, RuixueLi, YingqiuSun, Guijin...
    11页
    查看更多>>摘要：The objective of this study was to investigate protective effects of tree peony seed protein hydrolysate by Alcalase (AL-TPSPH) on oxidative damage, inflammation and apoptosis using Cd-induced zebrafish embryos. Zebrafish embryos were treated with either Cd (2 mu g/L) or AL-TPSPH (25, 50 and 75 mu g/mL) alone or in combination of both from 4 to 144 h post fertilization (hpf). The effects of these treatments on developments, antioxidant parameters and mRNA expression of genes related to oxidative damage, inflammation and apoptosis were examined. The results showed that co-treatment with Cd and AL-TPSPH significantly increased hatching and survival rates and decreased malformation rates of zebrafish embryos compared with Cd treatment alone group (P < 0.05). Cd-induced increase of MDA content, decreases of T-AOC content, GSH/GSSG ratio and activities of SOD, CAT and GPx in zebrafish embryos were modified upon treatment with AL-TPSPH. AL-TPSPH treatment significantly suppressed Cd-induced down-regulations of the antioxidant gene expressions (Mn-sod, Cat and GPx1a) in zebrafish embryos (P < 0.05). AL-TPSPH also prevented Cd-induced up-regulations of pro-inflammatory cytokine (TNF-alpha, IL-1 beta and IFN-gamma) expressions. Moreover, AL-TPSPH inhibited Cd-induced upregulations of pro-apoptotic genes (C-jun, Caspase-3 and Caspase-9) in zebrafish embryos. Collectively, these results indicated that AL-TPSPH could reduce Cd-induced oxidative damage, inflammation and apoptosis in zebrafish embryos, suggesting its future applications as functional food or pharmaceutical ingredient.
      原文链接:
    • NSTL
    • Elsevier

    Beneficial effects of re-feeding high alpha-linolenic acid diets on the muscle quality, cold temperature and disease resistance of tilapia

    Huang, XiaopingChen, FangGuan, JunfengXu, Chao...
    8页
    查看更多>>摘要：To investigate the effects of re-feeding high a-linolenic acid (ALA) diets on the muscle quality, cold temperature and disease resistance of the tilapia with nutritional history of soybean oil diets, three experimental diets with linoleic aicd (LA)/ALA ratios at 9 (D1, taking soybean oil as lipid sources), 3 and 1 (D2 and D3, taking soybean and linseed oils as lipid sources) were prepared to re-feed juveniles for 10 weeks, and the growth performance, muscle quality were analyzed. After the re-feeding trial, the fish were fasted for 8 weeks at cold temperature (15 degrees C-20 degrees C) and then subjected to the Aeromonas hydrophila challenge, and the cold temperature and disease resistance of the fish were evaluated. It was shown that a comparable growth performance was detected among the three dietary groups, while, the high feed efficiency and low viscerosomatic and hepatosomatic index were detected in the D2 and D3 groups compared with the D1 group. In addition, the docosahexaenoic acid (DHA) and n-3 polyunsaturated fatty acid (PUFA) levels of the muscle increased in a parallel pattern with the dietary ALA levels, and the muscular tenderness, adhesiveness, and chewiness were modified substantially in fish fed the diets D2 and D3. After 8-week fasting at cold temperature, the low serum total antioxidant capacity (T-AOC) and malondialdehyde (MDA) levels, and high serum lysozyme (LZM) and C3 levels were observed in the D2 and D3 groups compared with the D1 group. It was also shown that high eicosapentaenoic acid (EPA), DHA, and n-3 PUFA levels were observed in the intestine, liver, and spleen of fish from D2 and D3 groups. Correspondingly, in the fish of the D2 and D3 groups, the mRNA levels of lzm in the liver, intestine, and spleen, and c3 in the intestine and spleen were increased, while the mRNA levels of il-1 beta, ifn-gamma, and tnf-alpha in the intestine, and ifn-gamma, tnf-alpha in the liver, as well as spleen il-1 beta, were decreased. Furthermore, the survival at day 15 post-challenge of A. hydrophila in the D2 and D3 groups were higher than those of the D1 group. The results demonstrated that re-feeding high ALA diets were beneficial to the muscle quality, cold temperature and disease resistance in the tilapia, and provide a basis for selecting the dietary lipid sources of tilapia pre-winter feed.
      原文链接:
    • NSTL
    • Elsevier

    Response of cecropin transgenesis to challenge with Edwardsiella ictaluri in channel catfish Ictalurus punctatus

    Abass, Nermeen Y.Simora, Rhoda Mae C.Wang, JinhaiLi, Shangjia...
    7页
    查看更多>>摘要：Constructs bearing the cecropin B gene from the moth Hyalophora cecropia, driven by the cytomegalovirus (CMV) promoter, or the common carp beta-actin (beta-actin) promoter were transferred to channel catfish, Ictalurus punctatus via electroporation. One F-3 channel catfish family transgenic for cecropin transgene driven by the CMV promoter, and one F-1 channel catfish family transgenic for cecropin transgene driven by the common carp beta-actin promoter were produced. F-3 and F-1 individuals exhibited enhanced disease resistance when challenged in tanks with Edwardsiella ictaluri, the causative agent of enteric septicemia of catfish (ESC). Inheritance of the transgene by the F-1 and F-3 generation was 15% and 60%, respectively. Growth rates of the cecropin transgenic and non-transgenic full siblings (controls) channel catfish were not different (P > 0.05). All transgenic fish showed significant resistance to infection by ESC at day 3 and day 4 post exposure (P = 0.005). No correlation was detected between body weight and time to death for all genetic groups (P = 0.34). Results of our study confirmed that genetic enhancement of E. ictaluri resistance can be achieved by cecropin transgenesis in channel catfish. In addition to survival rate, improving survival time is essential because the extension of survival time gives a better chance to apply treatments to stop the bacterial infection.
      原文链接:
    • NSTL
    • Elsevier