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Extracellular vesicles and citrullination signatures are novel biomarkers in sturgeon (Acipenser gueldenstaedtii) during chronic stress due to seasonal temperature challenge 细胞外囊泡和瓜氨酸化特征是鲟鱼(Acipenser gueldenstaedtii)在季节性温度挑战导致的慢性应激期间的新型生物标志物。
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-18 DOI: 10.1016/j.fsi.2024.109974
Ana María Ferreira , Valeria Silva-Álvarez , Igor Kraev , Pinar Uysal-Onganer , Sigrun Lange
Acipenser gueldenstaedtii is one of the most cultured sturgeon species worldwide and of considerable economic value for caviar production. There are though considerable challenges around chronic stress responses due to increased summer temperatures, impacting sturgeons’ immune responses and their susceptibility to opportunistic infections. The identification of molecular and cellular pathways involved in stress responses may contribute to identifying novel biomarkers reflective of fish health status, crucial for successful sturgeon aquaculture. Protein citrullination is a calcium-catalysed post-translational modification caused by peptidylarginine deiminases (PADs), altering target protein function and affecting protein interactions in physiological and pathobiological processes. PADs can also modulate extracellular vesicle (EVs) profiles, which play critical roles in cellular communication, via transport of their cargoes (proteins, including post-translationally modified proteins, genetic material and micro-RNAs).
This study identified differences in EV signatures, and citrullinated proteins in sera from winter and summer farmed sturegeons. EVs were significantly elevated in sera of the summer chronically stressed group. The citrullinated proteins and associated gene ontology (GO) pathways in sera and serum-EVs of chronically heat stressed A. gueldenstaedtii, showed some changes, with specific citrullinated serum protein targets including alpa-2-macroglobulin, alpha globin, calcium-dependent secretion activator, ceruloplasmin, chemokine XC receptor, complement C3 isoforms, complement C9, plectin, selenoprotein and vitellogenin. In serum-EVs, citrullinated protein cargoes identified only in the chronically stressed summer group included alpha-1-antiproteinase, apolipoprotein B-100, microtubule actin crosslinking factor and histone H3. Biological gene ontology (GO) pathways related to citrullinated serum proteins in the chronically stressed group were associated with innate and adaptive immune responses, stress responses and metabolic processes. In serum-EVs of the heat-stressed group the citrullinome associated with various metabolic GO pathways.
In addition to modified citrullinated protein content, Serum-EVs from the stressed summer group showed significantly increased levels of the inflammatory associated miR-155 and the hypoxia-associated miR-210, but significantly reduced levels of the growth-associated miR-206.
Our findings highlight roles for protein citrullination and EV signatures in response to chronic heat stress in A. gueldenstaedtii, indicating a trade-off in immunity versus growth and may be of value for sturgeon aquaculture.
鲟鱼(Acipenser gueldenstaedtii)是世界上养殖最多的鲟鱼品种之一,在鱼子酱生产方面具有相当高的经济价值。然而,由于夏季气温升高,鲟鱼的免疫反应和对机会性感染的易感性受到影响,因此围绕慢性应激反应的研究面临着相当大的挑战。鉴定参与应激反应的分子和细胞途径可能有助于鉴定反映鱼类健康状况的新型生物标志物,这对鲟鱼的成功养殖至关重要。蛋白质瓜氨酸化是肽基精氨酸脱氨酶(PADs)催化的一种钙翻译后修饰,可改变目标蛋白质的功能,影响蛋白质在生理和病理生物学过程中的相互作用。PADs还能通过运输货物(蛋白质,包括翻译后修饰的蛋白质、遗传物质和微RNAs)来调节细胞外囊泡(EVs)特征,而EVs在细胞通讯中发挥着关键作用。这项研究发现了冬季和夏季养殖的鲟鱼血清中 EV 特征和瓜氨酸蛋白的差异。在夏季慢性应激组的血清中,EVs明显升高。瓜氨酸化蛋白和相关基因本体(GO)通路在长期热应激的鲟鱼血清和血清-EV中发生了一些变化,特定的瓜氨酸化血清蛋白靶标包括 alpa-2-巨球蛋白、甲型球蛋白、钙依赖性分泌激活因子、脑磷脂蛋白、趋化因子 XC 受体、补体 C3 同工型、补体 C9、褶皱蛋白、硒蛋白和卵黄原蛋白。在血清-EV中,仅在长期受压的夏季组中发现了瓜氨酸蛋白货物,包括甲型-1-抗蛋白酶、脂蛋白B-100、微管肌动蛋白交联因子和组蛋白H3。长期应激组中与瓜氨酸化血清蛋白相关的生物基因本体(GO)通路与先天性和适应性免疫反应、应激反应和代谢过程有关。在热应激组的血清-EV中,瓜氨酸化血清蛋白组与各种代谢GO通路有关。除了改变的瓜氨酸化蛋白含量外,夏季应激组的血清-EV中与炎症相关的miR-155和与缺氧相关的miR-210的水平显著增加,但与生长相关的miR-206的水平显著降低。我们的研究结果突显了瓜氨酸化蛋白和EV特征在鲟鱼对慢性热应激反应中的作用,表明了免疫与生长之间的权衡,可能对鲟鱼养殖有价值。
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引用次数: 0
Effects of dietary succinic acid supplementation on growth performance, digestive ability, intestinal development and immunity of large yellow croaker (Larimichthys crocea) larvae 补充琥珀酸对大黄鱼幼体生长性能、消化能力、肠道发育和免疫力的影响
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-17 DOI: 10.1016/j.fsi.2024.109972
Chenxiang Zhang , Yongtao Liu , Zhijie Shi , Chuanwei Yao , Jianmin Zhang , Yuntao Wang , Jiahui Liu , Kangsen Mai , Qinghui Ai
The application of artificial micro-diet is an effective way to improve and standardize the quality of aquatic animal larvae. However, the widespread adoption of micro-diet faces a bottleneck due to the limited utilization capacity of the larvae. A 30-day feeding experiment was carried out to investigate the effect of dietary succinic acid (SA) on the growth performance, digestive ability, intestinal development, and immunity of large yellow croaker larvae (initial body weight 11.33 ± 0.57 mg). Four isonitrogenous and isolipidic diets were formulated, incorporating 0.00 %, 0.01 %, 0.02 % and 0.03 % SA separately. The results showed that a diet with 0.02 % SA significantly increased both the final body weight and the specific growth rate of the larvae. Regarding digestive ability, 0.01 % SA supplementation significantly enhanced trypsin activity in both intestinal and pancreatic segments. In addition, 0.01 % SA supplementation notably improved amylase activity in the intestinal segment, while diets with 0.01%–0.02 % SA significantly improved lipase activity in the pancreatic segment. In terms of intestinal development, 0.01 % SA supplementation remarkably boosted the activities of alkaline-phosphatase and leucine-aminopeptidase on brush border membrane in intestine. Furthermore, 0.03 % SA supplementation significantly increased the expression of occludin. In terms of immunity, larvae fed diets with 0.01%–0.02 % SA exhibited significantly higher lysozyme activity compared to the control group. Supplementation with 0.01 % SA also significantly increased both iNOS activity and NO content. In summary, the findings of this study suggested that supplementing 0.02 % SA can improve the growth performance of large yellow croaker larvae by improving digestive enzymes activities, promoting intestinal development, and enhancing nonspecific immunity.
人工微量饲料的应用是提高和规范水生动物幼体质量的有效途径。然而,由于幼体的利用能力有限,微量饲料的广泛应用面临瓶颈。为了研究日粮琥珀酸(SA)对大黄鱼幼体(初始体重 11.33 ± 0.57 mg)生长性能、消化能力、肠道发育和免疫力的影响,我们进行了为期 30 天的饲养实验。分别添加 0.00%、0.01%、0.02% 和 0.03% 的 SA,配制了四种等氮和离脂日粮。结果表明,添加 0.02% SA 的日粮能显著提高幼虫的最终体重和特定生长率。在消化能力方面,添加 0.01% SA 能显著提高肠道和胰腺中胰蛋白酶的活性。此外,添加 0.01% SA 能显著提高肠道部分的淀粉酶活性,而添加 0.01%-0.02% SA 的日粮则能显著提高胰腺部分的脂肪酶活性。在肠道发育方面,添加 0.01% SA 能显著提高肠道刷状缘膜上碱性磷酸酶和亮氨酸氨肽酶的活性。此外,补充 0.03% 的 SA 还能显著提高闭塞素的表达。在免疫方面,与对照组相比,饲喂 0.01%-0.02% SA 日粮的幼虫溶菌酶活性明显提高。添加 0.01% SA 还能显著提高 iNOS 活性和 NO 含量。综上所述,本研究结果表明,添加 0.02% SA 可通过提高消化酶活性、促进肠道发育和增强非特异性免疫力来改善大黄鱼幼体的生长性能。
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引用次数: 0
Understanding the probiotic potential of Lactobacillus plantarum: Antioxidant capacity, non-specific immunity and intestinal microbiota improvement effects on Manila clam Ruditapes philippinarum 了解植物乳杆菌的益生潜力:对马尼拉蛤蜊 Ruditapes philippinarum 的抗氧化能力、非特异性免疫力和肠道微生物群的改善作用。
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-16 DOI: 10.1016/j.fsi.2024.109971
Longzhen Liu , Haonan Zhuang , Xiangli Tian , Yujia Zhou , Fangyi Wang , Zirong Liu , Jiamin Li , Minghui Jiao , Suyan Xue , Jiaqi Li , Weiwei Jiang , Yuze Mao
Lactic acid bacteria (LAB) have beneficial effects on aquatic animals, improving their immune system and intestinal microbiota. Nevertheless, the probiotic effects of LAB on the Manila clam Ruditapes philippinarum remain poorly understood. Herein, the effects of administering Lactobacillus plantarum at final doses of 1 × 105 CFU/L (T5 group), 1 × 107 CFU/L (T7 group), and 1 × 109 CFU/L (T9 group) in the rearing water for eight weeks were evaluated for the antioxidant capacity, non-specific immunity, resistance to Vibrio parahaemolyticus infection, and intestinal microbiota of R. philippinarum. The rearing water without the addition of L. plantarum served as a control. The results showed that the T7 and T9 groups demonstrated a significant elevation in the disease resistance of clams against V. parahaemolyticus, in the activities of alkaline phosphatase and lysozyme in the hepatopancreas, and in the expression of antioxidant- and immune-related genes, including SOD, GPx, and GST. Meanwhile, the T7 group showed a significant enhancement in superoxide dismutase and catalase activities and CAT expression, while the T9 group experienced a remarkable elevation in reduced glutathione content. Only catalase activity was markedly elevated in the T5 group. The expression of SOD, CAT, GPx, and GST was significantly elevated in three treatment groups following the V. parahaemolyticus challenge. The T7 group exhibited a significant increase in intestinal microbiota richness. Significant increases were noted in Firmicutes abundance across all three treatment groups and in Actinobacteriota in the T5 and T7 groups. Additionally, the opportunistic pathogen Escherichia-Shigella abundance significantly decreased in three treatment groups. Furthermore, administration of 1 × 107 CFU/L L. plantarum enhanced the stability of the intestinal microecosystem, whereas a dose of 1 × 109 CFU/L might have a negative effect. The application of three doses of L. plantarum significantly enhanced intestinal microbiota functions related to the immune response and oxidative stress regulation, while a higher dose (1 × 109 CFU/L) might inhibit several functions. In conclusion, the application of L. plantarum in the rearing water exerted beneficial effects on the antioxidant capacity, non-specific immunity, resistance to V. parahaemolyticus, and the intestinal microbiota stability and functions of R. philippinarum. The beneficial effects of L. plantarum on R. philippinarum were dose-dependent, and the final dose of 1 × 107 CFU/L exhibited the optimal effects.
乳酸菌(LAB)对水生动物有益,能改善它们的免疫系统和肠道微生物群。然而,人们对植物乳酸菌对马尼拉蛤蜊(Ruditapes philippinarum)的益生菌作用仍然知之甚少。本文评估了在饲养水中添加最终剂量为 1 × 105 CFU/L(T5 组)、1 × 107 CFU/L(T7 组)和 1 × 109 CFU/L(T9 组)的植物乳杆菌八周对菲利宾氏蛤抗氧化能力、非特异性免疫、抗副溶血性弧菌感染和肠道微生物群的影响。不添加植物乳杆菌的饲养水作为对照。结果表明,T7 和 T9 组的蛤蜊对副溶血性弧菌的抗病能力、肝胰脏中碱性磷酸酶和溶菌酶的活性以及抗氧化和免疫相关基因(包括 SOD、GPx 和 GST)的表达均有显著提高。同时,T7 组的超氧化物歧化酶、过氧化氢酶活性和 CAT 表达量显著提高,而 T9 组的还原型谷胱甘肽含量显著增加。只有过氧化氢酶活性在 T5 组有明显提高。副溶血性弧菌感染后,三个处理组的 SOD、CAT、GPx 和 GST 表达量均显著增加。T7 组的肠道微生物群丰富度明显增加。所有三个处理组的固有菌群和 T5 和 T7 组的放线菌群数量都有显著增加。此外,机会致病菌埃希氏菌在三个治疗组中的数量显著减少。此外,施用 1 × 107 CFU/L 的植物乳杆菌可增强肠道微生态系统的稳定性,而 1 × 109 CFU/L 的剂量则可能会产生负面影响。应用三种剂量的植物乳杆菌可显著增强肠道微生物群与免疫反应和氧化应激调节相关的功能,而较高剂量(1 × 109 CFU/L)可能会抑制多种功能。总之,在饲养水中添加植物乳杆菌对菲利宾纳氏鲤的抗氧化能力、非特异性免疫力、对副溶血性弧菌的抵抗力以及肠道微生物群的稳定性和功能都有益处。植物乳杆菌对菲利宾纳氏菌的有益作用与剂量有关,最终剂量为 1 × 107 CFU/L 时效果最佳。
{"title":"Understanding the probiotic potential of Lactobacillus plantarum: Antioxidant capacity, non-specific immunity and intestinal microbiota improvement effects on Manila clam Ruditapes philippinarum","authors":"Longzhen Liu ,&nbsp;Haonan Zhuang ,&nbsp;Xiangli Tian ,&nbsp;Yujia Zhou ,&nbsp;Fangyi Wang ,&nbsp;Zirong Liu ,&nbsp;Jiamin Li ,&nbsp;Minghui Jiao ,&nbsp;Suyan Xue ,&nbsp;Jiaqi Li ,&nbsp;Weiwei Jiang ,&nbsp;Yuze Mao","doi":"10.1016/j.fsi.2024.109971","DOIUrl":"10.1016/j.fsi.2024.109971","url":null,"abstract":"<div><div>Lactic acid bacteria (LAB) have beneficial effects on aquatic animals, improving their immune system and intestinal microbiota. Nevertheless, the probiotic effects of LAB on the Manila clam <em>Ruditapes philippinarum</em> remain poorly understood. Herein, the effects of administering <em>Lactobacillus plantarum</em> at final doses of 1 × 10<sup>5</sup> CFU/L (T5 group), 1 × 10<sup>7</sup> CFU/L (T7 group), and 1 × 10<sup>9</sup> CFU/L (T9 group) in the rearing water for eight weeks were evaluated for the antioxidant capacity, non-specific immunity, resistance to <em>Vibrio parahaemolyticus</em> infection, and intestinal microbiota of <em>R</em>. <em>philippinarum</em>. The rearing water without the addition of <em>L. plantarum</em> served as a control. The results showed that the T7 and T9 groups demonstrated a significant elevation in the disease resistance of clams against <em>V</em>. <em>parahaemolyticus</em>, in the activities of alkaline phosphatase and lysozyme in the hepatopancreas, and in the expression of antioxidant- and immune-related genes, including <em>SOD</em>, <em>GPx</em>, and <em>GST</em>. Meanwhile, the T7 group showed a significant enhancement in superoxide dismutase and catalase activities and <em>CAT</em> expression, while the T9 group experienced a remarkable elevation in reduced glutathione content. Only catalase activity was markedly elevated in the T5 group. The expression of <em>SOD</em>, <em>CAT</em>, <em>GPx</em>, and <em>GST</em> was significantly elevated in three treatment groups following the <em>V. parahaemolyticus</em> challenge. The T7 group exhibited a significant increase in intestinal microbiota richness. Significant increases were noted in Firmicutes abundance across all three treatment groups and in Actinobacteriota in the T5 and T7 groups. Additionally, the opportunistic pathogen <em>Escherichia-Shigella</em> abundance significantly decreased in three treatment groups. Furthermore, administration of 1 × 10<sup>7</sup> CFU/L <em>L. plantarum</em> enhanced the stability of the intestinal microecosystem, whereas a dose of 1 × 10<sup>9</sup> CFU/L might have a negative effect. The application of three doses of <em>L. plantarum</em> significantly enhanced intestinal microbiota functions related to the immune response and oxidative stress regulation, while a higher dose (1 × 10<sup>9</sup> CFU/L) might inhibit several functions. In conclusion, the application of <em>L. plantarum</em> in the rearing water exerted beneficial effects on the antioxidant capacity, non-specific immunity, resistance to <em>V</em>. <em>parahaemolyticus</em>, and the intestinal microbiota stability and functions of <em>R</em>. <em>philippinarum</em>. The beneficial effects of <em>L. plantarum</em> on <em>R</em>. <em>philippinarum</em> were dose-dependent, and the final dose of 1 × 10<sup>7</sup> CFU/L exhibited the optimal effects.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109971"},"PeriodicalIF":4.1,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142461339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Escherichia coli heat-labile enterotoxin B subunit as an adjuvant of mucosal immune combined with GCRV-II VP6 triggers innate immunity and enhances adaptive immune responses following oral vaccination of grass carp (Ctenopharyngodon idella) 大肠杆菌热嗜性肠毒素 B 亚基作为粘膜免疫佐剂与 GCRV-II VP6 结合使用可触发先天性免疫并增强草鱼口服疫苗后的适应性免疫反应
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-16 DOI: 10.1016/j.fsi.2024.109969
Jiyuan Yin , Huiliang Wu , Wei Li , Yingying Wang , Yingying Li , Xubing Mo , Siming Li , Yan Ren , Houjun Pan , Peng Jiang , Qing Wang
The grass carp reovirus (GCRV) is the most major pathogen that has threatened the grass carp (Ctenopharyngodon idella) industry of China for years. Though the oral vaccine has many advantages, the current vaccines still do not provide complete protection. Therefor the exploration of new preventive strategies is urgently needed. In this study, heat-labile enterotoxin B subunit of Escherichia coli (LTB) was combined with VP6 from GCRV type II (GCRV-II) via Lactococcus lactis expression system to form a potent oral vaccine and determines if fusion of LTB to the protective vaccine antigen can enhance protection in the fish. The expression of recombinant protein was confirmed by Western-blotting and enzyme-linked immunosorbent assay. The rare minnow was set as the model for the evaluation of the experiment administrated orally. The immune response including the antibody titer and the immune-related gene expression, and the protective efficacy which included the virus loaded and the relative protection, were thoroughly investigated after the trial. The results indicated that LTB can significantly elicit a higher neutralizing antibody responses and enhanced T-cell priming, activities and proliferation in mononuclear cells from intestine, spleen and kidney tissues when compared to the VP6 vaccine alone. Moreover, the combined adjuvant can significantly up-regulate type I interferon signaling in different immune organs, especially the mucosa associated lymphoidtissue which could not be induced by VP6 along, result in the contribution of the improvement in adaptive immune responses of the fish. In addition, challenge study showed that LTB combined VP6 could greatly improve the relative percent survival of the fish during the virus infection. These results highlight that LTB has the potential value to be a mucosal adjuvant of the fish, approaching for improving the efficacy of vaccination against GCRV-II, which does elicit both non-specific and specific immune responses.
草鱼再病毒(GCRV)是多年来威胁中国草鱼产业的最主要病原体。虽然口服疫苗有很多优点,但目前的疫苗仍不能提供完全的保护。因此,迫切需要探索新的预防策略。本研究通过乳酸乳球菌表达系统将大肠杆菌的热嗜性肠毒素 B 亚基(LTB)与 GCRV II 型(GCRV-II)的 VP6 结合形成强效口服疫苗,并确定 LTB 与保护性疫苗抗原融合是否能增强鱼类的保护力。通过 Western-blotting 和酶联免疫吸附试验证实了重组蛋白的表达。口服实验以珍稀鱲鱼为模型进行评估。试验后,对包括抗体滴度和免疫相关基因表达在内的免疫反应,以及包括病毒载量和相对保护率在内的保护效力进行了深入研究。结果表明,与单独接种 VP6 疫苗相比,LTB 能显著激发肠道、脾脏和肾脏组织单核细胞产生更高的中和抗体反应,并增强 T 细胞的启动、活性和增殖。此外,联合佐剂还能显著上调不同免疫器官的 I 型干扰素信号,尤其是 VP6 疫苗无法诱导的粘膜相关淋巴组织,从而有助于改善鱼类的适应性免疫反应。此外,挑战研究表明,LTB 联合 VP6 可大大提高鱼类在病毒感染期间的相对存活率。这些结果突出表明,LTB 具有作为鱼类粘膜佐剂的潜在价值,可用于提高 GCRV-II 疫苗接种的效果,因为 GCRV-II 疫苗可引起非特异性和特异性免疫反应。
{"title":"Escherichia coli heat-labile enterotoxin B subunit as an adjuvant of mucosal immune combined with GCRV-II VP6 triggers innate immunity and enhances adaptive immune responses following oral vaccination of grass carp (Ctenopharyngodon idella)","authors":"Jiyuan Yin ,&nbsp;Huiliang Wu ,&nbsp;Wei Li ,&nbsp;Yingying Wang ,&nbsp;Yingying Li ,&nbsp;Xubing Mo ,&nbsp;Siming Li ,&nbsp;Yan Ren ,&nbsp;Houjun Pan ,&nbsp;Peng Jiang ,&nbsp;Qing Wang","doi":"10.1016/j.fsi.2024.109969","DOIUrl":"10.1016/j.fsi.2024.109969","url":null,"abstract":"<div><div>The grass carp reovirus (GCRV) is the most major pathogen that has threatened the grass carp (<em>Ctenopharyngodon idella</em>) industry of China for years. Though the oral vaccine has many advantages, the current vaccines still do not provide complete protection. Therefor the exploration of new preventive strategies is urgently needed. In this study, heat-labile enterotoxin B subunit of <em>Escherichia coli</em> (LTB) was combined with VP6 from GCRV type II (GCRV-II) via <em>Lactococcus lactis</em> expression system to form a potent oral vaccine and determines if fusion of LTB to the protective vaccine antigen can enhance protection in the fish. The expression of recombinant protein was confirmed by Western-blotting and enzyme-linked immunosorbent assay. The rare minnow was set as the model for the evaluation of the experiment administrated orally. The immune response including the antibody titer and the immune-related gene expression, and the protective efficacy which included the virus loaded and the relative protection, were thoroughly investigated after the trial. The results indicated that LTB can significantly elicit a higher neutralizing antibody responses and enhanced T-cell priming, activities and proliferation in mononuclear cells from intestine, spleen and kidney tissues when compared to the VP6 vaccine alone. Moreover, the combined adjuvant can significantly up-regulate type I interferon signaling in different immune organs, especially the mucosa associated lymphoidtissue which could not be induced by VP6 along, result in the contribution of the improvement in adaptive immune responses of the fish. In addition, challenge study showed that LTB combined VP6 could greatly improve the relative percent survival of the fish during the virus infection. These results highlight that LTB has the potential value to be a mucosal adjuvant of the fish, approaching for improving the efficacy of vaccination against GCRV-II, which does elicit both non-specific and specific immune responses.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109969"},"PeriodicalIF":4.1,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142446559","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
bcIRF5 activates bcTBK1 phosphorylation to enhance PANoptosis during GCRV infection bcIRF5 激活 bcTBK1 磷酸化,以增强 GCRV 感染期间的 PAN 细胞凋亡
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-15 DOI: 10.1016/j.fsi.2024.109968
Can Yang , Jinwei Gao , Hao Wu , Zhenzhen Xiong , Jun Xiao , Yanfang Wu , Qing Yang , Zhonggui Xie , Rui Song , Dongsheng Ou , Hao Feng
TBK1 is an important IFN antiviral signalling factor, and in previous work black carp TBK1 (bcTBK1) and black carp IRF5 (bcIRF5) together promoted cell death in GCRV-infected cells. In this research, bcTBK1 and bcIRF5 were investigated both in vivo and in vitro to delineate their individual and combined functions. This study demonstrated that both bcTBK1 and bcIRF5 expressions were modulated in response to GCRV infection across the intestine, gill, kidney and spleen. In bcgill cells, overexpression of bcTBK1 and bcIRF5 initially suppressed the expression of cell death-related genes, including RIPK1, caspase1, caspase3 and bax, but this suppression was negated upon GCRV infection. In vivo, mRNA expression levels of RIPK1 and related genes varied by tissue following bcTBK1 or bcIRF5 overexpression and GCRV infection. Notably, intracellular co-overexpression of bcTBK1 and bcIRF5 led to significant upregulation of caspase3, caspase1, bax, and IL1β, along with enhanced caspase3 activity post-GCRV infection. This co-expression correlated with higher survival rates in black carp during GCRV infection and increased caspase3 mRNA in the spleen and gills. Hematoxylin-eosin (HE) staining indicated disorganized spleen tissue and edematous, hyperplastic gill changes in co-transfected groups after infection. TUNEL staining of tissue sections showed that DNA breakage was significantly stronger in the co-transfected group than in the other groups during GCRV infection. Further phosphorylation experiments showed that bcIRF5 promoted phosphorylation modification of bcTBK1. Thus, these data suggest that bcIRF5 activates bcTBK1 by enhancing its phosphorylation and promotes PANoptosis in GCRV-infected cells.
TBK1是一种重要的IFN抗病毒信号因子,在之前的研究中,黑鲤TBK1(bcTBK1)和黑鲤IRF5(bcIRF5)共同促进了GCRV感染细胞的死亡。在这项研究中,对 bcTBK1 和 bcIRF5 进行了体内和体外研究,以确定它们的单独和联合功能。研究表明,bcTBK1 和 bcIRF5 的表达在肠道、鳃、肾脏和脾脏对 GCRV 感染的反应中均有调节。在bcgill细胞中,过表达bcTBK1和bcIRF5最初会抑制细胞死亡相关基因的表达,包括RIPK1、caspase1、caspase3和bax,但这种抑制作用在感染GCRV后被抵消。在体内,bcTBK1 或 bcIRF5 过表达和 GCRV 感染后,RIPK1 和相关基因的 mRNA 表达水平因组织而异。值得注意的是,bcTBK1和bcIRF5在细胞内的共重表达会导致caspase3、caspase1、bax和IL1β的显著上调,并在GCRV感染后增强caspase3的活性。这种共表达与黑鲤在 GCRV 感染期间存活率较高以及脾脏和鳃中 caspase3 mRNA 增加有关。血色素-伊红(HE)染色显示,共同转染组在感染后脾脏组织紊乱,鳃水肿、增生。组织切片的 TUNEL 染色显示,在 GCRV 感染期间,共转染组的 DNA 断裂明显强于其他组。进一步的磷酸化实验表明,bcIRF5 促进了 bcTBK1 的磷酸化修饰。因此,这些数据表明,bcIRF5 通过增强 bcTBK1 的磷酸化激活了 bcTBK1,并促进了 GCRV 感染细胞的泛凋亡。
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引用次数: 0
Identification, functional analysis of chitin-binding proteins and the association of its single nucleotide polymorphisms with Vibrio parahaemolyticus resistance in Penaeus vannamei 万年青中几丁质结合蛋白的鉴定、功能分析及其单核苷酸多态性与副溶血性弧菌抗性的关联。
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-15 DOI: 10.1016/j.fsi.2024.109966
Shuang-Shuang Luo , Xiu-Li Chen , Ai-Jin Wang , Qing-Yun Liu , Min Peng , Chun-Ling Yang , Di-Gang Zeng , Yong-Zhen Zhao , Huan-Ling Wang
Chitin-binding proteins (CBPs) play pivotal roles in numerous biological processes in arthropods, including growth, molting, reproduction, and immune defense. However, their function in the antibacterial immune defense of crustaceans remains relatively underexplored. In this study, twenty CBPs were identified and characterized in Penaeus vannamei. Expression profiling highlighted that the majority of CBPs were highly expressed in the intestine and hepatopancreas and responded to challenge by Vibrio parahaemolyticus. To explore the role of these CBPs in innate immunity, six CBPs (PvPrg4, PvKrtap16, PvPT-1a, PvPT-1b, PvExtensin and PvCP-AM1159) were selected for RNAi experiments. Silencing of only PvPrg4 and PvKrtap16 significantly decreased the cumulative mortality of V. parahaemolyticus-infected shrimp. Further studies demonstrated that inhibition of PvPrg4 and PvKrtap16 resulted in a marked upregulation of genes associated with the NF-κB and JAK-STAT signaling pathways, as well as antimicrobial peptides (AMPs), in both the intestine and hepatopancreas. These results collectively suggested that PvPrg4 and PvKrtap16 potentially promote V. parahaemolyticus invasion by negatively regulating the JAK-STAT and NF-κB pathways, thereby inhibiting the expression of AMPs. In addition, SNP analysis identified three SNPs in the exons of PvPrg4 that were significantly associated with tolerance to V. parahaemolyticus. Taken together, these findings are expected to assist in the molecular marker-assisted breeding of P. vannamei associated with anti-V. parahaemolyticus traits, as well as expand our understanding of CBP functions within the immune regulatory system of crustaceans.
几丁质结合蛋白(CBPs)在节肢动物的许多生物过程中发挥着关键作用,包括生长、蜕皮、繁殖和免疫防御。然而,它们在甲壳类动物抗菌免疫防御中的功能仍未得到充分探索。本研究在凡纳滨对20种CBPs进行了鉴定和表征。表达谱分析显示,大多数 CBPs 在肠道和肝胰腺中高度表达,并对副溶血性弧菌的挑战做出反应。为了探索这些 CBPs 在先天性免疫中的作用,我们选择了六个 CBPs(PvPrg4、PvKrtap16、PvPT-1a、PvPT-1b、PvExtensin 和 PvCP-AM1159)进行 RNAi 实验。仅抑制 PvPrg4 和 PvKrtap16 能显著降低副溶血性弧菌感染虾的累积死亡率。进一步的研究表明,抑制 PvPrg4 和 PvKrtap16 会导致肠道和肝胰腺中与 NF-κB 和 JAK-STAT 信号通路相关的基因以及抗菌肽(AMPs)明显上调。这些结果共同表明,PvPrg4 和 PvKrtap16 可通过负向调节 JAK-STAT 和 NF-κB 通路,从而抑制 AMPs 的表达,潜在地促进副溶血性弧菌的入侵。此外,SNP 分析发现 PvPrg4 外显子中的三个 SNP 与副溶血性弧菌耐受性显著相关。综上所述,这些发现有望帮助分子标记辅助培育具有抗副溶血病毒特性的凡纳米贝,并扩大我们对 CBP 在甲壳类免疫调节系统中功能的了解。
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引用次数: 0
Comparative analysis of T-cell immunity between Streptococcus agalactiae susceptible and resistant tilapia (Oreochromis niloticus) 赤链球菌易感罗非鱼和耐受罗非鱼 T 细胞免疫力的比较分析
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-15 DOI: 10.1016/j.fsi.2024.109967
Jiansong Zhang , Ming Geng , Jun Xiao , Liting Chen , Yi Cao , Kang Li , Jialong Yang , Xiumei Wei
Nile tilapia (Oreochromis niloticus) is one of the important economic fish species cultured worldwide. However, Streptococcus agalactiae has emerged as a significant bacterial threat, severely impacting the economy of tilapia industry. The immune response underlying the resistance of tilapia to S. agalactiae are not well understood, hindering the reasonable evaluation of breeding and the formulation of effective strategies. In this study, we investigated the differences in T-cell immunity between S. agalactiae-resistant and -susceptible tilapia. Compared with susceptible tilapia, resistant tilapia exhibited a higher percentage of T cells and BrdU+ T cells during infection, indicating a superior proliferative capacity. Whether infected or not, T cells from resistant fish demonstrated a greater ability to resist apoptosis. Additionally, T cell effector genes, including interleukin (IL)-2, interferon (IFN)-γ, perforin A, and granzyme B were expressed at higher levels in resistant tilapia after infection. Along with these T-cell immune responses, resistant fish showed more effective clearance of infection. Our study elucidates the T-cell immune responses in resistant tilapia, which may contribute to the high resistance of tilapia to S. agalactiae, and provide valuable theoretical references for the selection and evaluation of disease-resistant fish strains in the future.
尼罗罗非鱼(Oreochromis niloticus)是全球养殖的重要经济鱼类之一。然而,无乳链球菌已成为一种重要的细菌威胁,严重影响了罗非鱼产业的经济效益。罗非鱼对 S. agalactiae 产生抗性的免疫反应尚不十分清楚,这阻碍了对育种的合理评估和有效策略的制定。在这项研究中,我们调查了抗赤潮镰刀菌罗非鱼和易感罗非鱼的 T 细胞免疫差异。与易感罗非鱼相比,抗性罗非鱼在感染期间表现出更高比例的 T 细胞和 BrdU+ T 细胞,表明其具有更强的增殖能力。无论是否感染,抗性罗非鱼的 T 细胞都表现出更强的抗凋亡能力。此外,抗性罗非鱼的 T 细胞效应基因,包括白细胞介素 (IL)-2、干扰素 (IFN)-γ、穿孔素 A 和颗粒酶 B 在感染后的表达水平更高。伴随着这些 T 细胞免疫反应,抗性罗非鱼表现出更有效的感染清除能力。我们的研究阐明了抗性罗非鱼的T细胞免疫反应,这可能是罗非鱼对S. agalactiae具有高度抗性的原因之一,并为今后抗病鱼种的选择和评价提供了有价值的理论参考。
{"title":"Comparative analysis of T-cell immunity between Streptococcus agalactiae susceptible and resistant tilapia (Oreochromis niloticus)","authors":"Jiansong Zhang ,&nbsp;Ming Geng ,&nbsp;Jun Xiao ,&nbsp;Liting Chen ,&nbsp;Yi Cao ,&nbsp;Kang Li ,&nbsp;Jialong Yang ,&nbsp;Xiumei Wei","doi":"10.1016/j.fsi.2024.109967","DOIUrl":"10.1016/j.fsi.2024.109967","url":null,"abstract":"<div><div>Nile tilapia (<em>Oreochromis niloticus</em>) is one of the important economic fish species cultured worldwide. However, <em>Streptococcus agalactiae</em> has emerged as a significant bacterial threat, severely impacting the economy of tilapia industry. The immune response underlying the resistance of tilapia to <em>S. agalactiae</em> are not well understood, hindering the reasonable evaluation of breeding and the formulation of effective strategies. In this study, we investigated the differences in T-cell immunity between <em>S. agalactiae</em>-resistant and -susceptible tilapia. Compared with susceptible tilapia, resistant tilapia exhibited a higher percentage of T cells and BrdU<sup>+</sup> T cells during infection, indicating a superior proliferative capacity. Whether infected or not, T cells from resistant fish demonstrated a greater ability to resist apoptosis. Additionally, T cell effector genes, including <em>interleukin (IL)-2</em>, <em>interferon (IFN)-γ</em>, <em>perforin A</em>, and <em>granzyme B</em> were expressed at higher levels in resistant tilapia after infection. Along with these T-cell immune responses, resistant fish showed more effective clearance of infection. Our study elucidates the T-cell immune responses in resistant tilapia, which may contribute to the high resistance of tilapia to <em>S. agalactiae</em>, and provide valuable theoretical references for the selection and evaluation of disease-resistant fish strains in the future.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"154 ","pages":"Article 109967"},"PeriodicalIF":4.1,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142446558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
CBS/CSE mediated H2S production induced AMPs expression through Toll pathway in crabs with black gill syndrome CBS/CSE 介导的 H2S 生产通过 Toll 通路诱导黑鳃病螃蟹体内 AMPs 的表达
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-12 DOI: 10.1016/j.fsi.2024.109965
Ziyue Zhu , Xingkong Ma , Xiaohan Liu , Liangmin Zheng , Lihua Zhang , Xiaoling Dai , Hao Li , Zhaoqian Zhang , Bingyan Wang , Xin Huang , Jiachun Ge , Qian Ren
The occurrence of black gill syndrome (BGS) is a serious threat to the healthy culture of Eriocheir sinensis. Studying the innate immune ability of E. sinensis with BGS can help develop new strategies for disease prevention and treatment. Antimicrobial peptides (AMPs) have crucial roles in crustacean humoral immunity. In this study, we found that the expression levels of two antilipopolysaccharide factor (EsALF7 and EsALF-L), one Toll receptor 3 (EsToll3), and one Pelle kinase (EsPelle) were upregulated in E. sinensis with BGS. Moreover, ALFs expressions in E. sinensis with BGS were positively regulated by EsToll3 and EsPelle. The content of hydrogen sulfide (H2S) in the gills of E. sinensis with BGS was increased. Further studies showed that the expressions of cystathionine β-synthase (EsCBS) and cystathionine γ-lyase (EsCSE) in the gills of E. sinensis with BGS were upregulated, which positively regulate the production of H2S. Whether there was a correlation between the upregulation of ALFs expression and changes in H2S content? Further studies showed that 1) the expressions of EsToll3, EsPelle, EsALF7, and EsALF-L in the gills of E. sinensis were upregulated under H2S exposure and 2) the knockdown of EsCBS and EsCSE in E. sinensis reduced the transcriptions of EsToll3, EsPelle, EsALF7, and EsALF-L. To sum up, these findings suggest that upregulation of H2S content induced by CBS/CSE promotes the expression of ALFs through the Toll pathway in E. sinensis suffering from BGS.
黑鳃病(BGS)的发生严重威胁着中华鳖的健康养殖。研究中华鳖黑鳃综合征的先天免疫能力有助于开发新的疾病预防和治疗策略。抗菌肽(AMPs)在甲壳类体液免疫中起着至关重要的作用。本研究发现,两种抗脂多糖因子(EsALF7和EsALF-L)、一种Toll受体3(EsToll3)和一种Pelle激酶(EsPelle)的表达水平在患有BGS的中华鳖中上调。此外,EsToll3和EsPelle还能正向调节ALFs的表达。BGS的中华鲟鳃中硫化氢(H2S)含量增加。进一步研究表明,胱硫醚β-合成酶(EsCBS)和胱硫醚γ-赖氨酸酶(EsCSE)在BGS中华鳖鳃中的表达上调,对H2S的产生有正向调节作用。ALFs 表达的上调与 H2S 含量的变化是否相关?进一步的研究表明:1)H2S暴露下,EsToll3、EsPelle、EsALF7和EsALF-L在中华鳖鳃中的表达上调;2)敲除EsCBS和EsCSE会降低EsToll3、EsPelle、EsALF7和EsALF-L的转录。综上所述,这些研究结果表明,CBS/CSE诱导的H2S含量上调会通过Toll途径促进BGS中E. sinensis的ALFs表达。
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引用次数: 0
Two novel antimicrobial peptides P33-57 and mP168-187 from zebrafish showing potent antibacterial activities 来自斑马鱼的两种新型抗菌肽 P33-57 和 mP168-187 显示出强大的抗菌活性
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-11 DOI: 10.1016/j.fsi.2024.109950
Dongqiu Guo , Hao Wang , Jing He , Liqiao Zhang , Longxiao Liu , Xia Wang
It is known that overuse or abuse of antibiotics has undoubtedly accelerated the global antibiotic resistance crisis, and long-time use of antibiotics may have adverse effects on the health of animal, human, and ecosystem. Therefore, it is necessary to find antibiotic alternatives that can be used in aquaculture and are non-toxic to the human. Here we clearly demonstrated that both the PH and FYVE domain of Plekhf2 in zebrafish have antibacterial properties and can interact with PGN in this study. Therefore, we screened four candidate peptides from the two domains. It was demonstrated that P152-172 and P168-187 had no obvious antibacterial activities, while P33-57 and mP168-187 had strong antibacterial activities, which may be used as antimicrobial peptides. Additionally, transmission electron microscopy experiment revealed that P33-57 and mP168-187 can destroy the cell wall of bacteria, thereby directly killing bacteria. Importantly, it was found that P33-57 and mP168-187 had no hemolysis to red blood cells and lacked cytotoxicity. In summary, P33-57 and mP168-187 could be seen as antibacterial activity centers of PLEKHF2 and may be promising antimicrobial peptides to combat bacterial infections facing an antibiotic-resistance crisis.
众所周知,过度使用或滥用抗生素无疑加速了全球抗生素耐药性危机,而且长期使用抗生素可能会对动物、人类和生态系统的健康产生不利影响。因此,有必要寻找可用于水产养殖且对人类无毒的抗生素替代品。在本研究中,我们清楚地证明了斑马鱼Plekhf2的PH和FYVE结构域都具有抗菌特性,并能与PGN相互作用。因此,我们从这两个结构域中筛选出四种候选肽。结果表明,P152-172和P168-187没有明显的抗菌活性,而P33-57和mP168-187具有很强的抗菌活性,可作为抗菌肽使用。此外,透射电子显微镜实验显示,P33-57 和 mP168-187 能破坏细菌的细胞壁,从而直接杀死细菌。重要的是,实验还发现 P33-57 和 mP168-187 对红细胞没有溶血作用,也没有细胞毒性。总之,P33-57 和 mP168-187 可被视为 PLEKHF2 的抗菌活性中心,可能是抗击面临抗生素耐药性危机的细菌感染的有前途的抗菌肽。
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引用次数: 0
Decoding the role of HIF-1α in immunoregulation in Litopenaeus vannamei under hypoxic stress 解码缺氧胁迫下 HIF-1α 在万年虾免疫调节中的作用
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-11 DOI: 10.1016/j.fsi.2024.109962
Jia L. Men , Yi J. Xue , Ying Fu , Xue Bai , Xiao B. Wang , Hai L. Zhou
Hypoxia poses a significant challenge to aquatic organisms, especially Litopenaeus vannamei (L. vannamei), which play a vital role in the global aquaculture industry. Hypoxia-inducible factor 1α (HIF-1α) is a pivotal regulator of the organism's adaptation to hypoxic conditions. To understand of how HIF-1α affects the immunity of L. vannamei under hypoxic conditions, we conducted a thorough study involving various approaches. These included observing tissue morphology, analyzing the expression of immune-related genes, assessing the activities of immune-related enzymes, and exploring immune-related pathways. Our study revealed that RNA interference (RNAi)-mediated knockdown of HIF-1α markedly reduced HIF-1α expression in the gill (75–95 %), whereas the reduction ranged from 2 to 43 % in the hepatopancreas. Knockdown of HIF-1α resulted in increased damage to both gill and hepatopancreatic tissues in hypoxic conditions. Additionally, immune-related genes, including Astakine (AST), Hemocyanin (HC), and Ferritin (FT), as well as immune-related enzymes such as Acid Phosphatase (ACP), Alkaline Phosphatase (AKP), and Phenoloxidase (PO), exhibited intricate regulatory patterns in response to hypoxia stress following the knockdown of HIF-1α. Transcriptome analysis revealed that HIF-1α knockdown significantly impacts multiple signaling pathways, including the JAK-STAT signaling pathway, Th17 cell differentiation pathways, PI3K-Akt signaling pathway, ErbB signaling pathway, MAPK signaling pathway, chemokine signaling pathway, ribosomal pathways, apoptosis, lysosomes and arachidonic acid metabolism. These alterations disrupt the organism's immune balance and interfere with normal metabolic processes, potentially leading to various immune-related diseases. We speculate that the weakened immune response resulting from HIF-1 inhibition is due to the reduced metabolic capacity, and the existence of a direct regulatory relationship between them requires further exploration. This study greatly advances our understanding of the vital role that HIF-1α plays in regulating immune responses in shrimp under hypoxic conditions, thereby deepening our comprehension of this critical biological mechanism.
缺氧对水生生物,尤其是在全球水产养殖业中发挥重要作用的凡纳滨对虾(Litopenaeus vannamei,L. vannamei)构成了重大挑战。缺氧诱导因子 1α(HIF-1α)是生物适应缺氧条件的关键调节因子。为了了解缺氧条件下 HIF-1α 如何影响凡纳滨鲤的免疫力,我们采用了多种方法进行了深入研究。这些方法包括观察组织形态、分析免疫相关基因的表达、评估免疫相关酶的活性以及探索免疫相关途径。我们的研究发现,RNA干扰(RNAi)介导的HIF-1α基因敲除显著降低了HIF-1α在鳃中的表达量(75-95%),而在肝胰脏中的表达量则降低了2-43%。敲除 HIF-1α 导致缺氧条件下鳃和肝胰腺组织的损伤增加。此外,免疫相关基因,包括天冬氨酸(AST)、血蓝蛋白(HC)和铁蛋白(FT),以及免疫相关酶,如酸性磷酸酶(ACP)、碱性磷酸酶(AKP)和酚氧化酶(PO),在HIF-1α基因敲除后的缺氧应激反应中表现出复杂的调控模式。转录组分析表明,HIF-1α基因敲除会显著影响多种信号通路,包括JAK-STAT信号通路、Th17细胞分化通路、PI3K-Akt信号通路、ErbB信号通路、MAPK信号通路、趋化因子信号通路、核糖体通路、细胞凋亡、溶酶体和花生四烯酸代谢。这些改变破坏了机体的免疫平衡,干扰了正常的新陈代谢过程,可能导致各种与免疫相关的疾病。我们推测,抑制 HIF-1 导致的免疫反应减弱是由于新陈代谢能力降低所致,二者之间是否存在直接的调控关系还需要进一步探讨。这项研究极大地推动了我们对 HIF-1α 在缺氧条件下调节对虾免疫反应的重要作用的认识,从而加深了我们对这一关键生物学机制的理解。
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引用次数: 0
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Fish & shellfish immunology
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