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Chronic heat exposure fails to trigger lasting thermal imprinting in juvenile black rockfish (Sebastes schlegelii) revealed by growth, histological, and transcriptomic analyses 通过生长、组织学和转录组学分析揭示,慢性热暴露不能触发幼黑岩鱼(sebases schlegelii)持久的热印迹。
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-15 DOI: 10.1016/j.fsi.2025.111077
Qing Ma , Tianci Nie , Hao Sun , Yan He , Jingjie Hu
Global climate change increasingly exposes marine species to thermal extremes, posing significant challenges to their survival and development. Fish, as ectothermic organisms, are especially vulnerable, with body temperature and metabolic processes dictated by the external environment. This study investigated whether chronic high-temperature exposure (27 °C) during early juvenile stages imposes a lasting “thermal imprint” on black rockfish (Sebastes schlegelii) once temperatures return to ambient conditions. Fish were tracked from 26 days post-parturition (dpp) through a 60-day exposure to elevated temperature and a subsequent 200-day period of normal rearing. Growth performance, muscle histology, and transcriptome-wide gene expression were comprehensively analyzed. Although elevated temperature initially suppressed growth metrics and elicited short-term transcriptomic changes related to myogenesis, stress response, and metabolic processes, these effects dissipated over time and did not persist once the fish reacclimated to ambient conditions. Consistent with these molecular findings, muscle histology showed no irreversible structural damage, despite a transient shift from hyperplastic to hypertrophic muscle fiber growth under heat stress. Together, these data indicate that chronic exposure to 27 °C in juvenile black rockfish does not result in long-term thermal imprinting. These findings improve our understanding of the species’ resilience to sub-lethal heat challenges and inform best practices for cultivating black rockfish under conditions of increasing climatic variability.
全球气候变化使海洋物种越来越多地面临极端温度,对其生存和发展构成重大挑战。鱼类,作为恒温生物,特别脆弱,体温和代谢过程由外部环境决定。本研究调查了幼鱼早期长期高温暴露(27°C)是否会在温度恢复到环境条件后对黑岩鱼(sebases schlegelii)产生持久的“热印记”。鱼从产后26天(dpp)开始被跟踪,经过60天的高温暴露和随后的200天正常饲养。综合分析生长性能、肌肉组织学和转录组基因表达。虽然升高的温度最初抑制了生长指标,并引发了与肌肉发生、应激反应和代谢过程相关的短期转录组变化,但这些影响随着时间的推移而消散,一旦鱼类重新适应环境条件,这些影响就不会持续存在。与这些分子发现一致,肌肉组织学显示,尽管在热应激下肌肉纤维生长从增生到肥厚的短暂转变,但没有不可逆的结构损伤。综上所述,这些数据表明,黑岩鱼幼鱼长期暴露在27°C环境中不会产生长期的热印记。这些发现提高了我们对该物种对亚致死热挑战的适应能力的理解,并为在气候日益变化的条件下养殖黑岩鱼提供了最佳实践。
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引用次数: 0
The specifically enhanced immune response of oyster Crassostrea gigas against the secondary encountered same pathogen 牡蛎长牡蛎对次生病原体的特异性增强免疫反应也遇到了同样的病原体。
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-15 DOI: 10.1016/j.fsi.2025.111073
Jiajun Zuo , Weilin Wang , Pan Luo , Lunyu Shen , Jiadi Wang , Jie Fan , Xiaoxu Zhou , Lingling Wang , Xiaodong Li , Linsheng Song
The innate immune cells in invertebrates can mount an enhanced immune response upon re-exposure to the same pathogen, which is referred to ‘immune priming’, similar to the ‘trained immunity’ in vertebrates. However, whether this enhanced immune effects during immune priming is pathogen-specific remains contradictory. In the present study, the specificity of immune response was investigated in the Pacific oyster (Crassostrea gigas) which received pre-stimulation with either Vibrio splendidus or Staphylococcus aureus followed by a secondary challenge with V. splendidus. After the secondary stimulation with V. splendidus, phagocytic activity of haemocytes against V. splendidus, rather than against S. aureus, was significantly higher in the V. splendidus-primed group compared to that of S. aureus-primed group, while there was no significant difference in the phagocytic activity of haemocytes against S. aureus or fluorescent microspheres. The mRNA expression levels of CgβGBP2, CgC1qDC-8, CgRab24, CgBigdefensin1 and CgBigdefensin2 and nitric oxide (NO) level in haemocytes all significantly up-regulated in the oyster primed with V. splendidus compared with those of S. aureus-primed group. The lysozyme levels in the hepatopancreas and cell-free haemolymph in the V. splendidus-primed group were also increased, concomitant with stronger inhibitory effect of cell-free haemolymph on the growth of V. splendidus than that of S. aureus-primed group. Moreover, both of the first V. splendidus and S. aureus stimulation significantly increased histone modification levels including H3K4me1, H3K4me3, H3K9ac and H3K27ac. These results indicated that pre-stimulation with a microbe could specifically induce enhanced cellular and humoral immunity in oyster C. gigas upon re-exposure, which provided a fresh idea for understanding the innate immune memory in invertebrates.
无脊椎动物的先天免疫细胞在再次暴露于同一病原体时可以产生增强的免疫反应,这被称为“免疫启动”,类似于脊椎动物的“训练免疫”。然而,在免疫启动过程中这种增强的免疫效应是否具有病原体特异性仍然存在矛盾。在本研究中,研究了太平洋牡蛎(长牡蛎)免疫反应的特异性,该牡蛎接受了脾弧菌或金黄色葡萄球菌的预刺激,然后接受了脾弧菌的二次攻击。脾弧菌二次刺激后,脾弧菌引物组的血细胞对脾弧菌的吞噬活性明显高于金黄色葡萄球菌引物组,而对金黄色葡萄球菌和荧光微球的吞噬活性无显著差异。与金黄色葡萄球菌引物组相比,脾弧菌引物处理的牡蛎细胞中CgβGBP2、CgC1qDC-8、CgRab24、CgBigdefensin1、CgBigdefensin2 mRNA表达水平和一氧化氮(NO)水平均显著上调。脾弧菌引物组肝胰脏和无细胞血淋巴溶菌酶水平均升高,且无细胞血淋巴对脾弧菌生长的抑制作用强于金黄色葡萄球菌引物组。此外,第一次脾弧菌和金黄色葡萄球菌刺激均显著提高组蛋白修饰水平,包括H3K4me1、H3K4me3、H3K9ac和H3K27ac。这些结果表明,微生物预刺激可特异性诱导牡蛎C. gigas再次暴露后的细胞和体液免疫增强,这为理解无脊椎动物的先天免疫记忆提供了新的思路。
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引用次数: 0
CXCL12a, an antimicrobial chemokine, contains a C-terminal helical structural fragment that exerts potent antimicrobial activity in largemouth bass (Micropterus salmoides) CXCL12a是一种抗菌趋化因子,含有c端螺旋结构片段,在大口黑鲈(Micropterus salmoides)中具有有效的抗菌活性。
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-15 DOI: 10.1016/j.fsi.2025.111075
Yanqi Zhang , Yangxi Xiang , Yazhen Hu , Chenghua Li
CXCL12 is a broad-spectrum antimicrobial protein across diverse species and plays a pivotal role in innate immunity. However, the specific structure underlying this activity remain poorly characterized. In this study, we reported that largemouth bass (Micropterus salmoides) encodes an evolutionarily conserved CXCL12a, which kills bacteria in a dose-dependent manner. MsCXCL12a directly kill bacteria through bacterial cell membrane disruption, cytoplasmic leakage and subsequent cell death. Notably, MsCXCL12a also exhibits calcium-independent bacterial aggregation property. By performing structure-charge analysis on the basis of its secondary structure, we identified that the C-terminal peptide of MsCXCL12a (MsCXCL12a55-75) retains broad-spectrum, stable antimicrobial activity while exhibiting negligible cytotoxicity. MsCXCL12a55-75 also kills bacteria by disrupting bacterial cell membrane integrity. Although MsCXCL12a55-75 retains the aggregation-mediating capability of MsCXCL12a, its aggregation efficiency is significantly reduced compared to MsCXCL12a. Using a largemouth bass model challenged with a lethal dose of Aeromonas hydrophila, MsCXCL12a and MsCXCL12a55-75 can substantially reduce tissue bacterial loads and mortality rates. These findings underscored the functional convergence between chemokine and antimicrobial peptide. The antimicrobial efficacy of MsCXCL12a55-75 in the fish model suggested its potential as a novel therapeutic candidate for bacterial infections, warranting further investigation into its translational applications.
CXCL12是一种广谱抗菌蛋白,存在于多种物种中,在先天免疫中起关键作用。然而,这种活动背后的具体结构仍然不清楚。在这项研究中,我们报道了大口黑鲈(Micropterus salmoides)编码一个进化上保守的CXCL12a,它以剂量依赖的方式杀死细菌。MsCXCL12a通过细菌细胞膜破裂、细胞质渗漏和随后的细胞死亡直接杀死细菌。值得注意的是,MsCXCL12a也表现出不依赖钙的细菌聚集特性。通过对其二级结构的结构电荷分析,我们发现MsCXCL12a的c端肽(MsCXCL12a55-75)保持了广谱、稳定的抗菌活性,而细胞毒性可以忽略不计。MsCXCL12a55-75还通过破坏细菌细胞膜的完整性来杀死细菌。虽然MsCXCL12a55-75保留了MsCXCL12a的聚合中介能力,但其聚合效率与MsCXCL12a相比明显降低。MsCXCL12a和MsCXCL12a55-75在嗜水气单胞菌致死剂量攻毒的大口黑鲈模型中可以显著降低组织细菌负荷和死亡率。这些发现强调了趋化因子和抗菌肽之间的功能趋同。MsCXCL12a55-75在鱼类模型中的抗菌效果表明,它有可能成为一种新的细菌感染治疗候选药物,值得进一步研究其转化应用。
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引用次数: 0
Efficacy of a novel oral bivalent vaccine with fucoidan as adjuvant against Aeromonas hydrophila and Edwardsiella tarda infections in Nile tilapia aquaculture 以岩藻糖聚糖为佐剂的新型口服二价疫苗对尼罗罗非鱼养殖中嗜水气单胞菌和迟发爱德华氏菌感染的防治效果。
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-13 DOI: 10.1016/j.fsi.2025.111074
Ritam Guha , Sreeja Lakshmi , Torben Krebs , Verena Jung Schroers , Mikolaj Adamek , Preetham Elumalai
The emerging diseases caused by Aeromonas hydrophila and Edwardsiella tarda severely impact aquaculture and lead to economic losses. Vaccines, as a safe prophylaxis method, can only protect against specific infections. Therefore, the development of a multivalent vaccine is necessary to enhance protection and cost-effectiveness for sustainable aquaculture. In this study, a bivalent vaccine against A. hydrophila and E. tarda was developed using fucoidan as an adjuvant. The vaccines were administered via the oral route to assess their protective efficacy against challenges. A survival rate of 63 % and 73 % was observed in the orally vaccinated fish where fucoidan was utilized as an adjuvant. Other immunological parameters, such as serum lysozyme, catalase, superoxide dismutase, bactericidal activity, and total IgM production, were measured. TCRβ, IgM, MHCI, MHCII, IFNγ, and IL8 gene expressions were significantly altered in the vaccinated fish. The gut microbiome alterations were noticeable in the vaccinated fish. Although feed-based oral vaccines necessitate antigen stability to withstand degradation in the fish gut, fucoidan used as an adjuvant has enhanced vaccine efficacy in orally vaccinated groups, indicating the potential of fucoidan as a novel immunomodulator and vaccine adjuvant in aquaculture.
由嗜水气单胞菌和迟发爱德华菌引起的新发病害严重影响水产养殖,造成经济损失。疫苗作为一种安全的预防方法,只能预防特定的感染。因此,有必要开发多价疫苗,以加强可持续水产养殖的保护和成本效益。本研究以褐藻聚糖为佐剂,研制了一种抗嗜水单胞菌和慢速单胞菌的二价疫苗。通过口服途径接种疫苗,以评估其对挑战的保护功效。使用岩藻多糖作为佐剂口服接种的鱼的存活率分别为63%和73%。其他免疫学参数,如血清溶菌酶、过氧化氢酶、超氧化物歧化酶、杀菌活性和总IgM的产生,被测量。TCRβ、IgM、MHCI、MHCII、IFNγ和IL8基因的表达在接种后显著改变。在接种疫苗的鱼中,肠道微生物组的改变是明显的。尽管以饲料为基础的口服疫苗需要抗原稳定性来抵御鱼肠道中的降解,但在口服疫苗接种组中,作为佐剂使用的岩藻糖聚糖增强了疫苗的效力,这表明岩藻糖聚糖在水产养殖中作为一种新型免疫调节剂和疫苗佐剂的潜力。
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引用次数: 0
Genome-wide characterization of the interleukin gene family in Nile tilapia (Oreochromis niloticus) spleen: identification, expression dynamics, and regulatory mechanism during Streptococcus agalactiae infection 尼罗罗非鱼(Oreochromis niloticus)脾脏白细胞介素基因家族的全基因组特征:鉴定、表达动态和无乳链球菌感染的调节机制
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-12 DOI: 10.1016/j.fsi.2025.111072
Zhang Wang , Shaowei Huang , Hong Liu , Chenglin Tang , Zhigang Liu , Jianmeng Cao , Miao Wang , Mengmeng Yi , Maixin Lu , Xiaoli Ke
The spleen is the principal site of immune hematopoiesis and a major target of Streptococcus agalactiae (S. agalactiae) attack in Nile tilapia (Oreochromis niloticus). However, the immune-regulatory mechanisms employed by this organ against S. agalactiae remain unclear. In this study, integrated transcriptomic analysis of spleen tissue identified 27 genes—among them the core cytokines IL-6, CXCL3 and IL1R2—that responded to S. agalactiae challenge. Genome-wide mining subsequently revealed a total of 27 IL-family members in Nile tilapia. Synteny and chromosomal location analyses indicated that expansion of the family was associated with tandem and whole-genome duplication events. The expression levels of IL-1β, IL-8, CXCL3, and IL-34 were higher than those of other IL-family members in tilapia spleen. Following S. agalactiae infection, IL-6, IL-8, IL-8L, IL-12a, and IL-17D were significantly up-regulated, whereas CXCL3 was markedly down-regulated at 24 h post-infection (hpi). Functional enrichment analysis showed that the 27 genes that respond to S. agalactiae were over-represented in the IL-17 signaling pathway. Co-treatment of primary spleen cells with an IL-17A antagonist attenuated the S. agalactiae-induced up-regulation of IL-6, IL-8, and IL8-like. Bisulfite-sequencing PCR further revealed that putative CpG islands within the IL-17D promoter were demethylated in the spleen following S. agalactiae infection. Collectively, this study provides a theoretical framework for understanding IL-mediated immune regulation in the tilapia spleen during S. agalactiae infection.
脾脏是尼罗罗非鱼(Oreochromis niloticus)免疫造血的主要部位,也是无乳链球菌(S. agalactiae)攻击的主要目标。然而,该器官对无乳链球菌的免疫调节机制尚不清楚。在本研究中,通过对脾脏组织的整合转录组学分析,鉴定出27个基因,其中包括核心细胞因子IL-6、CXCL3和il1r2,这些基因对S. agalactiae的攻击有反应。全基因组挖掘随后在尼罗罗非鱼中发现了总共27个il家族成员。Synteny和染色体定位分析表明,家族的扩展与串联和全基因组复制事件有关。IL-1β、IL-8、CXCL3和IL-34在罗非鱼脾脏中的表达水平高于其他il家族成员。无乳链球菌感染后,IL-6、IL-8、IL-8L、IL-12a和IL-17D在感染后24小时显著上调,而CXCL3在感染后24小时显著下调。功能富集分析表明,在IL-17信号通路中,27个响应S. agalactiae的基因被过量表达。与IL-17A拮抗剂共同处理原代脾细胞可减弱无乳链球菌诱导的IL-6、IL-8和IL-8样蛋白上调。亚硫酸酯测序PCR进一步显示,在无乳链球菌感染后,脾脏中IL-17D启动子内假定的CpG岛被去甲基化。总之,本研究为了解无乳链球菌感染期间罗非鱼脾脏中il介导的免疫调节提供了理论框架。
{"title":"Genome-wide characterization of the interleukin gene family in Nile tilapia (Oreochromis niloticus) spleen: identification, expression dynamics, and regulatory mechanism during Streptococcus agalactiae infection","authors":"Zhang Wang ,&nbsp;Shaowei Huang ,&nbsp;Hong Liu ,&nbsp;Chenglin Tang ,&nbsp;Zhigang Liu ,&nbsp;Jianmeng Cao ,&nbsp;Miao Wang ,&nbsp;Mengmeng Yi ,&nbsp;Maixin Lu ,&nbsp;Xiaoli Ke","doi":"10.1016/j.fsi.2025.111072","DOIUrl":"10.1016/j.fsi.2025.111072","url":null,"abstract":"<div><div>The spleen is the principal site of immune hematopoiesis and a major target of <em>Streptococcus agalactiae</em> (<em>S. agalactiae</em>) attack in Nile tilapia (<em>Oreochromis niloticus</em>). However, the immune-regulatory mechanisms employed by this organ against <em>S. agalactiae</em> remain unclear. In this study, integrated transcriptomic analysis of spleen tissue identified 27 genes—among them the core cytokines <em>IL-6</em>, <em>CXCL3</em> and <em>IL1R2</em>—that responded to <em>S. agalactiae</em> challenge. Genome-wide mining subsequently revealed a total of 27 <em>IL</em>-family members in Nile tilapia. Synteny and chromosomal location analyses indicated that expansion of the family was associated with tandem and whole-genome duplication events. The expression levels of <em>IL-1β</em>, <em>IL-8</em>, <em>CXCL3</em>, and <em>IL-34</em> were higher than those of other <em>IL-</em>family members in tilapia spleen. Following <em>S. agalactiae</em> infection, <em>IL-6</em>, <em>IL-8</em>, <em>IL-8L</em>, <em>IL-12a</em>, and <em>IL-17D</em> were significantly up-regulated, whereas <em>CXCL3</em> was markedly down-regulated at 24 h post-infection (hpi). Functional enrichment analysis showed that the 27 genes that respond to <em>S. agalactiae</em> were over-represented in the IL-17 signaling pathway. Co-treatment of primary spleen cells with an IL-17A antagonist attenuated the <em>S. agalactiae</em>-induced up-regulation of <em>IL-6</em>, <em>IL-8</em>, and <em>IL8-like</em>. Bisulfite-sequencing PCR further revealed that putative CpG islands within the <em>IL-17D</em> promoter were demethylated in the spleen following <em>S. agalactiae</em> infection. Collectively, this study provides a theoretical framework for understanding <em>IL</em>-mediated immune regulation in the tilapia spleen during <em>S. agalactiae</em> infection.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"169 ","pages":"Article 111072"},"PeriodicalIF":3.9,"publicationDate":"2025-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145755573","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
Tumor necrosis factor receptor-associated factor 6 (TRAF6) regulates Aeromonas hydrophila-induced autophagy in yellow catfish (Pelteobagrus fulvidraco) 肿瘤坏死因子受体相关因子6 (TRAF6)调控嗜水气单胞菌诱导的黄颡鱼自噬。
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-11 DOI: 10.1016/j.fsi.2025.111066
Zhi-Wei Nie , Yuan Hu , Yun-Fei Dang , Jiong Chen
Tumor necrosis factor receptor-associated factor 6 (TRAF6) is a key adaptor protein and E3 ubiquitin ligase involved in immune signaling and autophagy regulation. However, its function in pathogen-induced autophagy in teleost fish remains poorly understood. In this study, a TRAF6 homolog (PfTRAF6) was identified and characterized from yellow catfish (Pelteobagrus fulvidraco). The open reading frame of PfTRAF6 (1617 bp) encodes a 538 amino acid protein containing a conserved Zn-RING domain, zinc finger-TRAF domain, and C-terminal MATH domain. Phylogenetic analysis revealed that PfTRAF6 clustered closely with TRAF6 from other catfish species. PfTRAF6 transcripts were ubiquitously expressed across examined tissues, with the highest level in blood. Following Aeromonas hydrophila infection, both mRNA and protein levels of PfTRAF6 were significantly upregulated in immune-related tissues, particularly in blood and spleen. RNA interference-mediated knockdown of PfTRAF6 in monocytes/macrophages (MO/MΦ) suppressed the conversion of LC3-I to LC3-II, decreased the mRNA expression of pro-inflammatory cytokines and phosphorylation of P62, and reduced the number of autophagosomes observed by transmission electron microscopy. These findings indicate that PfTRAF6 positively regulates A. hydrophila-induced autophagy in yellow catfish. This study provides the first evidence that TRAF6 modulates bacterial infection-triggered autophagy in teleosts, expanding the understanding of the conserved TRAF6-autophagy axis in vertebrate innate immunity.
肿瘤坏死因子受体相关因子6 (Tumor necrosis factor receptor-associated factor 6, TRAF6)是参与免疫信号传导和自噬调节的关键衔接蛋白和E3泛素连接酶。然而,它在硬骨鱼病原体诱导的自噬中的作用仍然知之甚少。本研究从黄颡鱼(Pelteobagrus fulvidraco)中鉴定并鉴定了一个TRAF6同源物(PfTRAF6)。PfTRAF6的开放阅读框(1617 bp)编码一个538个氨基酸的蛋白,包含保守的Zn-RING结构域、锌指- traf结构域和c端MATH结构域。系统发育分析表明,PfTRAF6与其他鲶鱼品种的TRAF6聚类密切。PfTRAF6转录本在检查的组织中普遍表达,在血液中表达水平最高。在嗜水气单胞菌感染后,PfTRAF6的mRNA和蛋白水平在免疫相关组织中,特别是在血液和脾脏中显著上调。RNA干扰介导的单核/巨噬细胞(MO/MΦ)中PfTRAF6的下调抑制了LC3-I向LC3-II的转化,降低了促炎细胞因子的mRNA表达和P62的磷酸化,透射电镜观察到自噬体的数量减少。这些结果表明,PfTRAF6正调控嗜水单胞菌诱导的黄颡鱼自噬。本研究提供了TRAF6调节硬骨鱼细菌感染引发的自噬的第一个证据,扩大了对脊椎动物先天免疫中保守的TRAF6自噬轴的理解。
{"title":"Tumor necrosis factor receptor-associated factor 6 (TRAF6) regulates Aeromonas hydrophila-induced autophagy in yellow catfish (Pelteobagrus fulvidraco)","authors":"Zhi-Wei Nie ,&nbsp;Yuan Hu ,&nbsp;Yun-Fei Dang ,&nbsp;Jiong Chen","doi":"10.1016/j.fsi.2025.111066","DOIUrl":"10.1016/j.fsi.2025.111066","url":null,"abstract":"<div><div>Tumor necrosis factor receptor-associated factor 6 (TRAF6) is a key adaptor protein and E3 ubiquitin ligase involved in immune signaling and autophagy regulation. However, its function in pathogen-induced autophagy in teleost fish remains poorly understood. In this study, a TRAF6 homolog (<em>PfTRAF6</em>) was identified and characterized from yellow catfish (<em>Pelteobagrus fulvidraco</em>). The open reading frame of <em>PfTRAF6</em> (1617 bp) encodes a 538 amino acid protein containing a conserved Zn-RING domain, zinc finger-TRAF domain, and C-terminal MATH domain. Phylogenetic analysis revealed that <em>PfTRAF6</em> clustered closely with <em>TRAF6</em> from other catfish species. <em>PfTRAF6</em> transcripts were ubiquitously expressed across examined tissues, with the highest level in blood. Following <em>Aeromonas hydrophila</em> infection, both mRNA and protein levels of PfTRAF6 were significantly upregulated in immune-related tissues, particularly in blood and spleen. RNA interference-mediated knockdown of <em>PfTRAF6</em> in monocytes/macrophages (MO/MΦ) suppressed the conversion of LC3-I to LC3-II, decreased the mRNA expression of pro-inflammatory cytokines and phosphorylation of P62, and reduced the number of autophagosomes observed by transmission electron microscopy. These findings indicate that <em>PfTRAF6</em> positively regulates <em>A. hydrophila</em>-induced autophagy in yellow catfish. This study provides the first evidence that TRAF6 modulates bacterial infection-triggered autophagy in teleosts, expanding the understanding of the conserved TRAF6-autophagy axis in vertebrate innate immunity.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"169 ","pages":"Article 111066"},"PeriodicalIF":3.9,"publicationDate":"2025-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145741735","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
Develop subunit vaccines against Nocardia seriolae in hybrid snakehead (Channa maculata ♀ × Channa argus ♂) using two resuscitation-promoting factors 利用两种促复苏因子对杂交黑鱼♀×黑鱼♂研制抗诺卡菌亚单位疫苗。
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-11 DOI: 10.1016/j.fsi.2025.111067
Deyu Ning , Huiyuan Yang , Yan Li , Yuhao Li , Qiangcai He , Yiming Wen , Liqun Xia
Fish nocardiosis is a bacterial disease epidemic in aquaculture, and Nocardia seriolae is its main pathogen. The prevention and treatment of N. seriolae infection face multiple challenges, such as the unclear mechanism of chronic infection, resistance to phagocyte killing, and unsatisfied therapeutic efficacy of antibiotics. Resuscitation-promoting factors (Rpfs) are widely found in gram-positive bacteria, and have been reported to be vaccine candidates against tuberculosis and nocardiosis. In this study, RpfB and RpfD of N. seriolae ZJ0503 were obtained by prokaryotic expression and combined with Freund's Incomplete Adjuvant (FIA) as subunit vaccines, respectively. Finally, their efficacy were accessed in hybrid snakehead. The results showed that the relative survival rates (RPS) of subunit vaccines RpfB + FIA and RpfD + FIA were 77.78 % and 70.37 %, respectively. Histopathological examination revealed that hybrid snakehead did not show cytotoxic activity after intraperitoneal injection of RpfB + FIA and RpfD + FIA vaccines. In addition, both RpfB + FIA and RpfD + FIA vaccines increased the activity of multiple nonspecific immune parameters such as LYZ, CAT, ACP, AKP and SOD in the fish serum. The production of specific antibodies (IgM) and the expression of immune-related genes (tnfα, il-1β, cd4, cd8α, mhciα and mhciiα) were significantly increased compared with the control groups. In summary, RpfB and RpfD subunit vaccines were safe and can effectively protect hybrid snakehead from N. seriolae infection, and are ideal vaccine candidates for preventing fish nocardiosis.
鱼类诺卡菌病是水产养殖中的一种细菌性传染病,其主要病原菌为锡勒诺卡菌。慢性感染机制尚不清楚、对吞噬细胞的杀伤有耐药性、抗生素治疗效果不理想等问题是预防和治疗锡雷奈瑟菌感染面临的多重挑战。复苏促进因子(Rpfs)广泛存在于革兰氏阳性细菌中,据报道是结核病和诺卡菌病的候选疫苗。本研究通过原核表达获得了serolae ZJ0503的RpfB和RpfD,并分别与Freund's Incomplete佐剂(FIA)联合作为亚单位疫苗。最后,在杂交蛇头身上进行药效试验。结果表明,亚单位疫苗RpfB+FIA和RpfD+FIA的相对存活率分别为77.78%和70.37%。组织病理学检查显示,腹腔注射RpfB+FIA和RpfD+FIA疫苗后,杂交蛇头未表现出细胞毒活性。此外,RpfB+FIA和RpfD+FIA疫苗均能提高鱼血清中LYZ、CAT、ACP、AKP和SOD等多种非特异性免疫参数的活性。特异性抗体(IgM)的产生和免疫相关基因(tnfα、il-1β、cd4、cd8α、mhciiα和mhciiα)的表达均显著高于对照组。综上所述,RpfB和RpfD亚单位疫苗是安全的,可以有效地保护杂交蛇头免受锡雷奈菌感染,是预防鱼诺卡菌病的理想候选疫苗。
{"title":"Develop subunit vaccines against Nocardia seriolae in hybrid snakehead (Channa maculata ♀ × Channa argus ♂) using two resuscitation-promoting factors","authors":"Deyu Ning ,&nbsp;Huiyuan Yang ,&nbsp;Yan Li ,&nbsp;Yuhao Li ,&nbsp;Qiangcai He ,&nbsp;Yiming Wen ,&nbsp;Liqun Xia","doi":"10.1016/j.fsi.2025.111067","DOIUrl":"10.1016/j.fsi.2025.111067","url":null,"abstract":"<div><div>Fish nocardiosis is a bacterial disease epidemic in aquaculture, and <em>Nocardia seriolae</em> is its main pathogen. The prevention and treatment of <em>N. seriolae</em> infection face multiple challenges, such as the unclear mechanism of chronic infection, resistance to phagocyte killing, and unsatisfied therapeutic efficacy of antibiotics. Resuscitation-promoting factors (Rpfs) are widely found in gram-positive bacteria, and have been reported to be vaccine candidates against tuberculosis and nocardiosis. In this study, RpfB and RpfD of <em>N. seriolae</em> ZJ0503 were obtained by prokaryotic expression and combined with Freund's Incomplete Adjuvant (FIA) as subunit vaccines, respectively. Finally, their efficacy were accessed in hybrid snakehead. The results showed that the relative survival rates (RPS) of subunit vaccines RpfB + FIA and RpfD + FIA were 77.78 % and 70.37 %, respectively. Histopathological examination revealed that hybrid snakehead did not show cytotoxic activity after intraperitoneal injection of RpfB + FIA and RpfD + FIA vaccines. In addition, both RpfB + FIA and RpfD + FIA vaccines increased the activity of multiple nonspecific immune parameters such as LYZ, CAT, ACP, AKP and SOD in the fish serum. The production of specific antibodies (IgM) and the expression of immune-related genes (<em>tnfα</em>, <em>il-1β</em>, <em>cd4</em>, <em>cd8α</em>, <em>mhciα</em> and <em>mhciiα</em>) were significantly increased compared with the control groups. In summary, RpfB and RpfD subunit vaccines were safe and can effectively protect hybrid snakehead from <em>N. seriolae</em> infection, and are ideal vaccine candidates for preventing fish nocardiosis.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"169 ","pages":"Article 111067"},"PeriodicalIF":3.9,"publicationDate":"2025-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145741712","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
Chronic low-dose nitrite exposure impairs gill health, immunity, and induces apoptosis in starry flounder (Platichthys stellatus) 慢性低剂量亚硝酸盐暴露损害星牙鲆的鳃健康、免疫并诱导细胞凋亡。
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-11 DOI: 10.1016/j.fsi.2025.111070
Jie Lian , Qing Ji , Bo Yang, Junnan Zhu, Mingyue Zhang, Suyue Zhou, Xingqiang Wang, Jiachen Yu
While acute nitrite toxicity in aquatic species is well-documented, the effects of chronic low-dose exposure remain poorly understood. This study investigates the effects of chronic (30-day) hidden (0.4 and 0.8 mg/L) nitrite exposure on the welfare of starry flounder (Platichthys stellatus) by examining histology, antioxidant responses, apoptosis, and immune function. Results indicate that prolonged nitrite exposure revealed dose-dependent gill damage, including swelling and structural changes, likely as a protective mechanism against nitrite uptake and oxidative damage. SOD, CAT, GSH and T-AOC decreased dose-dependently, while malondialdehyde (MDA) levels increased, indicating oxidative stress and lipid peroxidation. Apoptotic pathways were activated, as evidenced by transcriptional upregulation of Caspase-3/9 and dysregulation of the P53-Bax/Bcl-2 axis. The effective execution of this program was further supported by immunofluorescence, showing dose-dependent increases in both P53 and Caspase-3 protein signals. Immunofluorescence analysis demonstrated a corresponding dose-dependent increase in P53 and Caspase-3 protein, suggesting post-transcriptional modulation. TUNEL-positive signals corroborated these molecular changes. Immune responses exhibited pro-inflammatory dominance (the levels of TNF-α, IL-6, IL-8 and IL-1β were increased) alongside anti-inflammatory suppression (TGF-β and IL-10 was down-regulated). The downregulation of TGF-β was further confirmed at the protein level by immunofluorescence. Meanwhile, IL-1β the increase occurred in a time-dependent fashion in H group. Notably, HSP70/90 upregulation suggested nitrite-induced activation of stress-mediated immunomodulatory pathways. These findings highlight the complex physiological stress response to chronic nitrite exposure, emphasizing the need for improved aquaculture practices to safeguard fish welfare and product quality.
虽然对水生物种的急性亚硝酸盐毒性有充分的记录,但对慢性低剂量接触的影响仍然知之甚少。本研究通过观察星形比目鱼(Platichthys stellatus)的组织学、抗氧化反应、细胞凋亡和免疫功能,探讨了慢性(30 d)隐藏(0.4和0.8 mg/L)亚硝酸盐暴露对星形比目鱼健康的影响。结果表明,长时间的亚硝酸盐暴露显示出剂量依赖性的鳃损伤,包括肿胀和结构变化,可能是亚硝酸盐摄取和氧化损伤的保护机制。SOD、CAT、GSH和T-AOC呈剂量依赖性下降,而丙二醛(MDA)水平升高,表明氧化应激和脂质过氧化。凋亡通路被激活,如Caspase-3/9转录上调和P53-Bax/Bcl-2轴失调。免疫荧光进一步支持了该计划的有效执行,显示P53和Caspase-3蛋白信号的剂量依赖性增加。免疫荧光分析显示P53和Caspase-3蛋白的相应剂量依赖性增加,提示转录后调节。tunel阳性信号证实了这些分子变化。免疫反应表现出促炎优势(TNF-α、IL-6、IL-8和IL-1β水平升高)和抗炎抑制(TGF-β和IL-10下调)。免疫荧光进一步在蛋白水平证实TGF-β下调。同时,IL-1β在H组呈时间依赖性升高。值得注意的是,HSP70/90的上调表明亚硝酸盐诱导了应激介导的免疫调节途径的激活。这些发现突出了慢性亚硝酸盐暴露的复杂生理应激反应,强调需要改进水产养殖做法,以保障鱼类福利和产品质量。
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引用次数: 0
Cloning, tissue distribution, and stimulus-responsive mRNA expression of CD10 (neprilysin) in Lethenteron reissneri 雷氏肠虫CD10的克隆、组织分布及刺激响应性mRNA表达。
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-11 DOI: 10.1016/j.fsi.2025.111071
Liang Zhao , Zihao Yan , Xianhui Li , Ya Pang , Ding Li , Guoxiu Ma , Yuetong Zhao , Mingwei Du , Feng Sun , Yinglun Han
This study presents the first systematic characterization of the molecular evolution and immune-related expression of the Lr-CD10 in the Lethenteron reissneri. Screening and bioinformatic analysis of the L. reissneri genome database confirmed that Lr-CD10 retains the core functional domains characteristic of the vertebrate CD10 gene family, including the zinc-binding motif HEXXH and catalytic active site. This indicates that the metallopeptidase activity of CD10 was established within a conserved functional framework early in vertebrate evolution. To investigate the potential biological role of Lr-CD10, we performed targeted gene knockdown in L. reissneri and conducted comparative transcriptome analysis, generating comprehensive gene expression profiles. The results suggest that CD10-mediated metallopeptidase activity may play dual roles: inflammation regulation and extracellular matrix remodeling. Our findings reveal an immune regulatory network associated with CD10 in an early vertebrate and provide transcriptomic evidence supporting its multifaceted functional evolution. Furthermore, cross-species comparison indicates that certain core immune regulatory modules are conserved across evolution, although their specific functional roles may diverge. This work provides a novel theoretical framework for investigating the evolutionary-developmental principles of the immune system and proposes an ancient origin for tumor microenvironment regulation, offering testable hypotheses for future functional studies.
本研究首次系统表征了莱氏肠虫(Lethenteron reissneri)中Lr-CD10的分子进化和免疫相关表达。L. reissneri基因组数据库的筛选和生物信息学分析证实,Lr-CD10保留了脊椎动物CD10基因家族的核心功能域特征,包括锌结合基元HEXXH和催化活性位点。这表明CD10的金属肽酶活性是在脊椎动物进化早期的保守功能框架内建立的。为了研究Lr-CD10的潜在生物学作用,我们在L. reissneri中进行了靶向基因敲低,并进行了比较转录组分析,获得了全面的基因表达谱。结果表明,cd10介导的金属肽酶活性可能具有炎症调节和细胞外基质重塑的双重作用。我们的研究结果揭示了早期脊椎动物中与CD10相关的免疫调节网络,并提供转录组学证据支持其多方面的功能进化。此外,跨物种比较表明,某些核心免疫调节模块在整个进化过程中是保守的,尽管它们的具体功能作用可能不同。这项工作为研究免疫系统的进化发育原理提供了一个新的理论框架,并提出了肿瘤微环境调控的古老起源,为未来的功能研究提供了可测试的假设。
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引用次数: 0
Bacterial infection triggers CD3ε+ T cell expansion and interaction with macrophages in grass carp (Ctenopharyngodon idella) 细菌感染触发草鱼CD3ε+ T细胞扩增并与巨噬细胞相互作用。
IF 3.9 2区 农林科学 Q1 FISHERIES Pub Date : 2025-12-10 DOI: 10.1016/j.fsi.2025.111068
Wa Gao , Jing Cen , Liwen Zhu , Hui Yan , Yueshuang Ji , Jing Xu , Bangjie Wang , Yanwei Zhang , Jun Zou , Junya Wang
While T cells are well established as key players in mammalian adaptive immunity, their functional roles in teleosts remain poorly understood. Here, a monoclonal antibody against grass carp (Ctenopharyngodon idella) CD3ε was generated, and characterized by Western blotting, flow cytometry, and immunofluorescence. In healthy fish, CD3ε+ T cells accounted for 5.1 %, 4.0 %, and 2.6 % of splenic, head kidney, and peripheral blood lymphocytes, respectively. Upon Aeromonas hydrophila infection, CD3ε+ T cell populations elevated significantly in immune tissues such as spleen, head kidney, and posterior intestine, accompanied by upregulated transcription of T cell markers (Cd3ε, Tcr-β, and Lck). Strikingly, immunofluorescence analysis revealed that CD3ε+ T cells and MCSFR+ macrophages were located in the same region in the spleen tissue of A. hydrophila-infected fish, suggesting functional crosstalk between T cells and macrophages. Our findings provide the first evidence of T cell-macrophage interaction in teleost antibacterial defense, offering insights into the adaptive immune responses in fish.
虽然T细胞在哺乳动物适应性免疫中发挥着重要作用,但它们在硬骨鱼中的功能作用仍然知之甚少。本实验制备了一种针对草鱼CD3ε的单克隆抗体,并通过Western blotting、流式细胞术和免疫荧光技术对其进行了鉴定。在健康鱼类中,CD3ε+ T细胞分别占脾脏、头肾和外周血淋巴细胞的5.1%、4.0%和2.6%。在嗜水气单胞菌感染后,免疫组织如脾脏、头肾和后肠中的CD3ε+ T细胞群显著升高,并伴有T细胞标记物(CD3ε、Tcr-β和Lck)的转录上调。引人注目的是,免疫荧光分析显示,CD3ε+ T细胞和MCSFR+巨噬细胞位于嗜水单胞虫感染的鱼脾脏组织的同一区域,提示T细胞和巨噬细胞之间存在功能串扰。我们的发现提供了T细胞-巨噬细胞相互作用在硬骨鱼抗菌防御中的第一个证据,为鱼类的适应性免疫反应提供了见解。
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引用次数: 0
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Fish & shellfish immunology
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