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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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    The dynamic immune responses of Mandarin fish (Siniperca chuatsi) to ISKNV in early infection based on full-length transcriptome analysis and weighted gene co-expression network analysis

    Zhou Y.Fu H.-C.Wang Y.-Y.Huang H.-Z....
    15页
    查看更多>>摘要:? 2022Mandarin fish (Siniperca chuatsi) been seriously harmed by infectious spleen and kidney necrosis virus (ISKNV) in recent years, but the early immune response mechanism of infection is still unknown. Here, we performed RNA sequencing on the spleens of mandarin fish infected with ISKNV at 0, 12, 24, 48, and 72 h post-infection (hpi) using short-read Illumina RNA sequencing and long-read Pacific Biosciences isoform sequencing to generate a full-length transcriptome. The immune responses of mandarin fish infected with ISKNV at the molecular level were characterized by RNA-seq analysis and weighted gene co-expression network analysis (WGCNA). A total of 26,528 full-length transcript sequences were obtained. There were 2,729 (1,680 up-regulated and 1,112 down-regulated), 1,874 (1,136 up-regulated and 738 down-regulated), 2,032 (1,158 up-regulated and 847 down-regulated), and 4,176 (2,233 up-regulated and 1,943 down-regulated) differentially expressed genes (DEGs) in mandarin fish at 12, 24, 48, and 72 hpi, compared with uninfected fish, respectively. A total of four modules of co-expressed DEGs identified by WGCNA were significantly positively correlated to the four time points after infection, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the immune-related DEGs in all these modules were mainly enriched in Phagosome, Endocytosis, Herpes simplex infection, and Cytokine-cytokine receptor interaction pathways. Further analysis showed that oher signaling pathways, including CAMs, NOD-like receptor and ER protein processing, Intestinal immune network for IgA production, TLR pathway, and Apoptosis significantly enriched in four modules corresponding to 12, 24, 48, and 72 hpi respectively, had specifically participated in the immune response. Hub genes identified based on the high-degree nodes in the WGCN, including CAM3, IL-8, CCL21, STING, SNX1, PFR and TBK1, and some DEGs such as MHCI, MHCII, TfR, STING, TNF α, TBK1, IRF1, and NF-kB, BCR, IgA and Bcl-XL had involved in dynamic molecular response of mandarin fish to ISKNV infection. In sum, this study provides a set of full-length transcriptome of the spleen tissue of mandarin fish for the first time and revealed a group of immune genes and pathways involved in different temporal responses to ISKNV infection, which has implications for resource conservation and aiding the development of strategies to prevent virus early infection for mandarin fish.

    Hypoxia stress affects the physiological responses, apoptosis and innate immunity of Kuruma shrimp, Marsupenaeus japonicus

    Wang P.Zhao S.Yu S.Xie S....
    9页
    查看更多>>摘要:? 2022 Elsevier LtdFor commercial aquatic animals, hypoxia phenomenon often occurs in live transport and aquaculture. In previous studies, much interest has been focused on antioxidant enzyme activities and could not present the complexities. The multifaceted responses, especially considering physiological indexes, histological structure, cell apoptosis, and immune pathways, are still unknown. In this study, we investigated the comprehensive hypoxic responses of Marsupenaeus japonicus. The results showed that the physiological indexes showed time-dependent changes upon hypoxia stress. Hypoxia stress led to significant tissue damage and cell apoptosis in the gill and hepatopancreas. Compared with the control group, the apoptosis index (AI) of the 12 h hypoxic treatment increased significantly (p < 0.05) in the gills and hepatopancreas. Comparative transcriptome analysis identified 900 and 1400 differentially expressed genes (DEGs) in the gill and hepatopancreas, respectively. Several DEGs were related to the lysosome, glycolysis/gluconeogenesis, citrate cycle, and apoptosis, and seven of them were validated using quantitative real-time PCR. This study provided valuable clues to understanding the mechanisms underlying the hypoxic responses of M. japonicus.

    Dietary inclusion of rambutan (Nephelium lappaceum L.) seed to Nile tilapia (Oreochromis niloticus) reared in biofloc system: Impacts on growth, immunity, and immune-antioxidant gene expression

    Xuan C.L.Wannavijit S.Outama P.Lumsangkul C....
    10页
    查看更多>>摘要:? 2022 Elsevier LtdAn eight-week feeding trial was carried out to determine the effects of rambutan seed (RS) as a feed additive on the growth, skin mucus, serum immune parameters, and gene expression of Nile tilapia (Oreochromis niloticus) raised under a biofloc system. Nile tilapia fingerlings (14.77 ± 0.80 g fish?1) were fed five experimental diets containing 0, 5, 10, 20, and 40 g kg?1 of RS, corresponding to five treatments (RS0, RS5, RS10, RS20, and RS40) with three replications per treatment. The results showed that fish consuming the RS10 and RS20 diets presented a substantial (P < 0.05) improvement in specific growth rate (SGR), weight gain (WG), and feed conversion ratio (FCR) after eight weeks. The highest values were recorded in the RS10 diet; however, there were no significant (P > 0.05) differences exhibited in the fish survival rates between treatments. The RS supplementation diets demonstrated greater immunological parameters, particularly skin mucus and serum immune responses (P < 0.05), than that of the control after eight the eight-week feeding trial. The highest level was seen in fish fed the RS10; followed by the RS20, RS40 (P > 0.05), and RS5 diets. Regarding gene expressions, IL1, IL8, LBP, GSTa, and GSR genes were significantly up-regulated in fish provided the RS10 diet in comparison to the control and other supplemented diets (P < 0.05). However, no significant up-regulation was found in these genes among the RS0, RS5, RS20, and RS40 diets, with the exception of the GPX gene. Similarly, up-regulation of IL-8, LBP, GSTa, GPX, and GSR were noted in fish fed the RS10 diet (P < 0.05). Notably, no significant differences were evident in these genes among the RS5, RS20, and RS40 diets. In conclusion, fish fed RS10 (10 g kg?1) significantly enhanced growth, skin mucus, serum immunities, and immune-antioxidants related gene expressions of Nile tilapia raised under biofloc system.

    Paired miRNA and RNA sequencing provides a first insight into molecular defense mechanisms of Scapharca broughtonii during ostreid herpesvirus-1 infection

    Bai C.-M.Zhang X.Gu L.Li Y.-N....
    9页
    查看更多>>摘要:? 2022 Elsevier LtdOstreid herpesvirus 1 (OsHV-1) infection caused mortalities with relevant economic losses in bivalve aquaculture industry worldwide. Initially described as an oyster pathogen, OsHV-1 can infect other bivalve species, like the blood clam Scapharca broughtonii. However, at present, little is known about the molecular interactions during OsHV-1 infection in the blood clam. We produced paired miRNA and total RNA-seq data to investigate the blood clam transcriptional changes from 0 to 72 h after experimental infection with OsHV-1. High-throughput miRNA sequencing of 24 libraries revealed 580 conserved and 270 new blood clam miRNAs, whereas no genuine miRNA was identified for OsHV-1. Total 88–203 differently expressed miRNAs were identified per time point, mostly up-regulated and mainly targeting metabolic pathways. Most of the blood clam mRNAs, in contrast, were down-regulated up to 60 h post-injection, with the trend analysis revealing the activation of immune genes only when comparing the early and latest stage of infection. Taken together, paired short and long RNA data suggested a miRNA-mediated down-regulation of host metabolic and energetic processes as a possible antiviral strategy during early infection stages, whereas antiviral pathways appeared upregulated only at late infection.

    Immunomodulation and skin microbiota perturbations during an episode of chronic stress in gilthead seabream

    Camara-Ruiz M.Garcia-Beltran J.M.Esteban M.A.Cerezo I.M....
    12页
    查看更多>>摘要:? 2022 The AuthorsFish stress is a major concern in the aquaculture industry. Many stressors coming from routine practices can predispose fish to compromised growth, immunity and overall health. This study focuses on the characterization of the skin microbiota using next generation sequencing (NGS) platform by targeting a genomic marker 16S and to determine growth performance and immune status of gilthead seabream (Sparus aurata) during an episode of chronic stress. Two groups were established: control group and chronically stressed group. Stressed fish were subjected to 1 min air exposure twice a week for 4 weeks. Results showed that stress negatively affected fish growth performance. Cellular and humoral systemic immunity remained unaffected while local immunity in skin was positively stimulated (total IgM and peroxidase). Skin mucus microbial composition showed significant differences especially after 14 days. Stressed fish showed a decrease in the abundance of the genera Acinetobacter, NS3a_marine_group and Pseudomonas, while Pseudoalteromonas and Marinagarivorans increased significantly. In conclusion, air exposure stress was associated with alterations in skin mucosal immunity and microbial composition that may have been beneficial to the host favoring adaptation to stress.

    An integrin alpha 4 (ChIntα 4) from oyster Crassostrea hongkongensis mediates the hemocytes phagocytosis towards Vibrio alginolyticus

    Fu Y.-W.Chen W.-F.He M.-H.Tang L....
    11页
    查看更多>>摘要:? 2022 Elsevier LtdIntegrins, a family of cell adhesion transmembrane receptors, mediate cell adhesion, migration, proliferation, apoptosis, and phagocytosis. In the present study, an integrin ChIntα 4 from Crassostrea hongkongensis was characterized to investigate its role in defensing against pathogenic bacterium Vibrio alginolyticus. The full-length cDNA sequence of ChIntα 4 was 3572 bp with an open reading frame (ORF) of 3168 bp, which encoded a polypeptide with 1055 amino acids. The mRNA expression of ChIntα 4 in the hemocytes was significantly up-regulated at 6 h and 24 h post V. alginolyticus stimulation (p < 0.01). The recombinant ChIntα 4 protein could agglutinate the rabbit red blood cells and Gram-negative bacteria V. alginolyticus and Escherichia coli. Moreover, the phagocytic activity of the hemocytes was significantly down-regulated from 46.9% to 32.7% when blocked with anti-ChIntα 4 antibody, and it was significantly up-regulated from 42.7% to 59.5% post transfection with pCI-neo-ChIntα 4 plasmid (p < 0.05). In conclusion, these findings demonstrated that ChIntα 4 might be involved in resisting V. alginolyticus infection and regulating phagocytosis as a cell adhesion receptor in C. hongkongensis.

    Silencing of Nrf2 in Litopenaeus vannamei, decreased the antioxidant capacity, and increased apoptosis and autophagy

    Huang Y.Li Q.Yuan Y.Zhang Z....
    11页
    查看更多>>摘要:? 2022Nuclear factor E2-related factor 2 (Nrf2) is a multifunctional transcription factor that plays an important role in antioxidant activities. However, its effect on antioxidant capacity in Litopenaeus vannamei, an economically important crustacean, remains unclear. In this study, the role of Nrf2 in response to oxidative stress in L. vannamei was determined by its effect on relevant gene expression and enzymatic activity. Nrf2 was cloned and analyzed. Results revealed that Nrf2 contains a 1575 bp open reading frame encoding 524 amino acids and a conserved bZIP Maf domain. The sequence similarity of Nrf2 between L. vannamei and Homarus americanus is 81%. Although the Nrf2 expression was detected in all tissues, the Nrf2 expression levels were the highest in the hepatopancreas, followed by the eyestalk and muscle. RNA interference significantly decreased the expression of antioxidant-related genes (SOD, GPX, CAT, Trx, and HO-1; p < 0.05), significantly upregulated the expression of autophagy genes (Atg3, Atg4, Atg5, Atg10, and Atg12; p < 0.05) and apoptosis genes (Caspase-3 and P53; p < 0.05). Moreover, SOD, CAT, and GPX enzyme activities decreased whereas the MDA activity increased. The histological results of the shrimp injected with dsRNA-Nrf2 showed that the hepatic tubules were irregularly arranged, the lumen was abnormal, and a few hepatic tubules were significantly enlarged compared with those of the dsRNA-EGFP group. The hepatocytes were also vacuolated. In conclusion, this study provided evidence that Nrf2 is involved in the regulation of antioxidant capacity, oxidative stress, apoptosis, and autophagy in shrimp.

    Insulin-like signaling promotes limb regeneration in the Chinese mitten crab (Eriocheir sinensis)

    Tian Y.Li X.Zuo J.Zhao R....
    8页
    查看更多>>摘要:? 2022 Elsevier LtdIn the pond culture of Chinese mitten crabs, limb autotomy seriously affects the feeding efficiency, immunity and survival. Therefore, it is crucial to understand the mechanism of limb regeneration of mitten crabs, so that culture strategies could be developed to reduce the limb impairment rate. The insulin-like signaling (ILS) pathway is evolutionarily conserved, and plays key roles in the growth and immunity of various species. In this study, a full-length cDNA of insulin-like receptor (EsInR) was identified from Eriocheir sinensis, and its mRNA expression patterns during limb regeneration was evaluated. The cDNA of EsInR includes a 4326 bp ORF encoding a protein of 1441 amino acids, with conserved α-and β-subunits. The EsInR and genes related to ILS were found to be upregulated during limb regeneration, which indicated that ILS plays a key role in limb regeneration of E. sinensis. Our experiment revealed that inhibition of ILS through injection of the InR inhibitor GSK1838705A at the blastema formation stage significantly reduced the limb regeneration rate compared to control group. In addition, injection of GSK1838705A also reduced the size of newly formed limbs after the molting cycle. Furthermore, we found that genes related to myogenesis were downregulated following injection of InR inhibitor both before and after molting. The results also indicated that cyclins and CDK1 were downregulated, while CKIs were upregulated following treatment with the InR inhibitor. These results suggest that ILS regulates limb regeneration in E. sinensis by promoting muscle growth and regeneration in response to autotomy stress. Thus, we identified a conserved insulin-like receptor in E. sinensis, and provide new evidence for the involvement of ILS in the regulation of limb autotomy and regeneration in crustaceans.

    Comprehensive transcriptome analysis reveals the effect of feeding rhythm on the immunity and metabolism of Acipenser dabryanus

    Chen Y.Wu X.Lai J.Liu Y....
    12页
    查看更多>>摘要:? 2022 Elsevier LtdAcipenser dabryanus is a species endemic to Yangtze River drainage in China and is listed as a critical endangered species on the IUCN Red List. In the present study, the liver and spleen transcriptomes were analyzed by comparing the data of A. dabryanus that experienced nine different feeding rhythms (once a day diurnal, T1; two times a day diurnal, T2; three times a day diurnal, T3; four times a day, T4; five times a day, T5; six times a day, T6; once a day nocturnal, Tn1; two times a day nocturnal, Tn2; and three times a day nocturnal, Tn3). Transcriptome sequencing generated 1,901,236,482 clean reads, encompassing 570.4 Gb of sequence data. The reads were assembled into 287,372 unigenes with an average length of 803 bp and an N50 of 1004 bp. KEGG analysis showed that 1,080, 1,030, and 1216 unigenes were annotated to lipid metabolism, amino acid metabolism and carbohydrate metabolism, respectively, and 2549 unigenes were annotated to the immune system category. Differentially expressed genes (DEGs) between different feeding frequency groups or between nighttime and daytime feeding were obtained and functionally enriched. Importantly, DEGs participating in nutrition metabolism and various immunoregulation pathways and their expression profiles in A. dabryanus were discussed. Interestingly, the majority of key genes related to lipid metabolism or in immunodependent gene families, such as antimicrobial peptides, Toll-like receptors, chemokines, NOD-like receptors, B cell receptors and the major histocompatibility complex, were all significantly upregulated in animals in the T6 group compared to the characteristics of animals in the T2 group that had a normal feeding frequency. In addition, light/dark rhythm also affected the immunity of A. dabryanus, and fish fed at night possessed an improved immune response than fish fed at daytime. Our study suggested that feeding six times a day is optimal for A. dabryanus juvenile growth as it enhances the organism's nutrition metabolism and immune function.

    Expression of p38MAPK and its regulation of apoptosis under high temperature stress in the razor clam Sinonovacula constricta

    Wang Y.Han Y.Lv M.Li Y....
    10页
    查看更多>>摘要:? 2022p38MAPK is a key branch of the MAPK (mitogen-activated protein kinase) pathway that plays an important role in physiological processes such as apoptosis, cell proliferation and growth. In this experiment, we screened and identified one p38MAPK gene in the razor clam Sinonovacula constricta, which encoded 359 amino acids and was widely expressed in various adult tissues. After 24 h of high temperature stress at 34 °C, the transcript expression of p38MAPK showed significant changes in all tested tissues. In particular in the gill and hepatopancreas tissues, where the expression increased 1.81 and 7.83 times compared with the control group, respectively (P < 0.01). Furthermore, we examined the expression of the apoptosis suppressor gene Bcl-2 and pro-apoptosis gene Bax by knock-down of p38MAPK with dsRNA interference in the gill and hepatopancreas tissues. The obvious up-regulation expression of Bcl-2 and significant suppression of Bax were observed, respectively (P < 0.01). Moreover, the TUNEL staining technique was used to detect apoptosis before and after interference. The degree of apoptosis in the gill and hepatopancreas tissues was reduced after interference with p38MAPK, and the ROS content was significantly reduced (P < 0.01). The results suggested that p38MAPK had a regulatory role in the heat tolerance of razor clams.