Fish & shellfish immunology最新文献_第3页

Hepcidin defense patterns in intestine of mandarin fish (Siniperca chuatsi) against Aeromonas hydrophila infection 鳜鱼肠道对嗜水气单胞菌感染的肝素防御模式

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-02 DOI: 10.1016/j.fsi.2024.110003

Jin-hua Gao , Jin-liang Zhao , Ning-xin Guo , Temesgen Tola , Wen-bo Xue , Kang-ning Cai , Xiao-li Yao , Miao Hu

Bacterial septicemia caused by Aeromonas hydrophila is one of the common bacterial diseases in aquaculture. Antimicrobial peptides, including hepcidin, are key components of the innate immune system in fish, playing a role in defense against pathogens. This study investigated the defense patterns of hepcidin in mandarin fish (Siniperca chuatsi) following A. hydrophila infection using gene expression analysis and in vitro antibacterial assays. We measured changes in the expression of iron metabolism-related genes (hepcidin, fpn, ftn, tf, tfr1) and immune-related genes (il-1β, il-6, il-8, il-10, tnf-α, socs3, nkap, tlr1, tlr2) in the intestine post-infection. MBC experiment demonstrated that the hepcidin synthetic peptide has an inhibitory effect on the growth of V. parahaemolyticus (32 μg/ml), A. hydrophilus (64 μg/ml), and F. columnaris (128 μg/ml), but not E. tarda (>256 μg/ml). After A. hydrophilus challenge, fpn, tf and tfr1 with peak expression at 24 hpi (2.75), 12 hpi (4.43) and 6 hpi (7.41), respectively. Hepcidin and ftn expression was highest at 48 hpi (115.01) and 72 hpi (4.16). The Fe²⁺ content peaked at 6 hpi (2.64 μmol/l) and reached its lowest at 12 hpi (1.12 μmol/l) in the intestine. After pathogen challenge, il-1β, il-8, socs3, tlr2, and hepcidin showed trends of increased and then decreased, with peak expression at 72 hpi (5.13, 37.05, 3.08) and 48 hpi (5.35, 115.01), respectively. These findings suggested that hepcidin plays a key role in the defense against A. hydrophila: initially restricting bacterial growth through iron metabolism (0–48 hpi), and later modulating immune responses via the TNF (by inducing il-1β and socs3) and Toll-like receptor pathways (by inducing il-8 and tlr2) (48–96 hpi). This study provides novel insights into the immune function of hepcidin in fish and its potential application in managing bacterial infections in aquaculture.

由嗜水气单胞菌引起的细菌性败血症是水产养殖中常见的细菌性疾病之一。抗菌肽（包括肝磷脂酶）是鱼类先天性免疫系统的重要组成部分，在防御病原体方面发挥作用。本研究利用基因表达分析和体外抗菌试验研究了鳜鱼感染姬蛙后肝素的防御模式。我们测定了感染后肠道中铁代谢相关基因（hepcidin、ffn、ftn、tf、tfr1）和免疫相关基因（il-1β、il-6、il-8、il-10、tnf-α、socs3、nkap、tlr1、tlr2）的表达变化。MBC 实验表明，肝磷脂合成肽对副溶血性大肠杆菌（32 μg/ml）、嗜水气单胞菌（64 μg/ml）和柱孢痢疾杆菌（128 μg/ml）的生长有抑制作用，但对塔达氏杆菌（>256 μg/ml）没有抑制作用。蚜虫挑战后，fpn、tf 和 tfr1 分别在 24 hpi（2.75）、12 hpi（4.43）和 6 hpi（7.41）达到表达峰值。Hepcidin 和 ftn 在 48 hpi (115.01) 和 72 hpi (4.16) 表达量最高。肠道中的 Fe2+ 含量在 6 hpi 达到最高（2.64 μmol/l），在 12 hpi 达到最低（1.12 μmol/l）。病原体挑战后，il-1β、il-8、socs3、tlr2 和 hepcidin 的表达量呈先升高后降低的趋势，分别在 72 hpi（5.13、37.05、3.08）和 48 hpi（5.35、115.01）达到峰值。这些研究结果表明，血红素在防御嗜水甲藻的过程中发挥着关键作用：最初通过铁代谢限制细菌生长（0-48 hpi），随后通过 TNF（通过诱导 il-1β 和 socs3）和 Toll 样受体途径（通过诱导 il-8 和 tlr2）调节免疫反应（48-96 hpi）。这项研究为了解鱼类血红素的免疫功能及其在管理水产养殖中细菌感染的潜在应用提供了新的视角。

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引用次数: 0

Kinetics of viral hemorrhagic septicemia virus (VHSV) and immune response in olive flounder (Paralichthys olivaceus) at altered temperature 橄榄鲽（Paralichthys olivaceus）在改变温度条件下的病毒性出血性败血症病毒（VHSV）动力学和免疫反应。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-02 DOI: 10.1016/j.fsi.2024.110004

Yo-Seb Jang , Su-Young Yoon , Rahul Krishnan , Myung-Joo Oh

Viral hemorrhagic septicemia virus (VHSV) is a pathogen that causes hemorrhagic septicemia in olive flounder at water temperatures below 15 °C, leading to symptoms such as abdominal swelling due to ascites and muscle hemorrhaging, and in severe cases, mortality. In this study, we investigated the proliferation of NCCs, the transcriptional analysis of CD4 and CD8, and the expression of pro-inflammatory cytokines (IL-1β, IL6, TNFα) and the cytokines (IL12, IL15, IFN-1β, IFNγ) involved in cytotoxic cell activation in the kidney of olive flounder during VHSV infection at suboptimal temperature (17 °C) and following a shift to optimal temperature (10 °C). Following viral infection, the population of NCCs and CD8 gene expression steadily increased. Meanwhile, IL12, IL15, and IFN-1β levels exhibited an early surge, reaching their peak within 1–2 days post-challenge at 17 °C. Notably, in olive flounder that were re-challenged with VHSV at 10 °C after an initial challenge at 17 °C, there was a rapid and robust expansion of NCCs and increased CD4 and CD8 gene expression, demonstrating the mounting of immune memory by NCCs and an efficient adaptive immune response from CD4⁺ and CD8⁺ T cells against VHSV infection.

病毒性出血性败血症病毒（VHSV）是一种病原体，在水温低于15 °C时会引起橄榄鲽出血性败血症，导致腹水引起的腹部肿胀和肌肉出血等症状，严重时会导致死亡。在这项研究中，我们调查了橄榄鲽在次优温度（17 °C）下感染 VHSV 和转入最适温度（10 °C）后肾脏中 NCCs 的增殖、CD4 和 CD8 的转录分析、促炎细胞因子（IL-1β、IL6、TNFα）和参与细胞毒性细胞活化的细胞因子（IL12、IL15、IFN-1β、IFNγ）的表达。病毒感染后，NCCs 数量和 CD8 基因表达量稳步增加。与此同时，IL12、IL15 和 IFN-1β 的水平也出现了早期激增，在 17 °C条件下，在病毒感染后 1-2 天内达到峰值。值得注意的是，橄榄鲽在 17 °C首次受到 VHSV 病毒感染后，在 10 °C再次受到 VHSV 病毒感染，NCCs 快速而稳健地扩增，CD4 和 CD8 基因表达增加，这表明 NCCs 形成了免疫记忆，CD4+ 和 CD8+ T 细胞对 VHSV 病毒感染做出了有效的适应性免疫反应。

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引用次数: 0

Paralichthys olivaceus GSDME-mediated pyroptosis is regulated by multiple caspases in different manners 橄榄副鳞鱼 GSDME 介导的裂解受多种不同方式的 Caspases 调节。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-02 DOI: 10.1016/j.fsi.2024.110002

Kangwei Hao , Liming Yuan , Chao Yu , Hang Xu , Li Sun

Pyroptosis is a type of programmed cell death mediated by gasdermin (GSDM). GSDM is activated by caspase (CASP), which cleaves GSDM to release the N-terminal (NT) fragment that forms channels in the plasma membrane and leads to cell death. To date, research on pyroptosis in teleost is limited. In this study, we examined the activation and regulation mechanism of pyroptosis in flounder Paralichthys olivaceus. P. olivaceus gasdermin E (PoGSDME) was found to be cleaved by six P. olivaceus caspases (PoCASP1/3a/3b/7/8a/8b). PoCASP1/3a/3b/7 cleaved primarily at ₂₄₅FEAD₂₄₈, which generated an NT fragment (NT₂₄₈) that induced robust pyroptosis. PoCASP8a/8b cleaved both the full length PoGSDME and NT₂₄₈ at ₂₀₂IEKD₂₀₅, thus destroying the biological activity of PoGSDME and NT₂₄₈. Nine residues crucial for PoGSDME function were identified, of which, F2, L19, and G85 were essential to plasma membrane translocation. During bacterial infection, PoGSDME and PoCASP1 expressions were significantly upregulated in flounder tissues, and PoGSDME, as well as PoCASP1, activation occurred in flounder cells accompanied with the processing cleavage of IL-1β and IL-18. Together these results revealed both the activation and the inhibition mechanisms of GSDME-mediated pyroptosis in flounder, and added new insights into the regulation of pyroptosis in fish.

裂解病是一种由gasdermin（GSDM）介导的程序性细胞死亡。GSDM由Caspase（CASP）激活，Caspase会裂解GSDM，释放出N-末端（NT）片段，在质膜上形成通道，导致细胞死亡。迄今为止，有关远洋鱼类热凋亡的研究还很有限。在这项研究中，我们考察了比目鱼（Paralichthys olivaceus）热噬的激活和调控机制。研究发现，比目鱼的气蛋白 E（PoGSDME）可被比目鱼的六种 Caspases（PoCASP1/3a/3b/7/8a/8b）裂解。PoCASP1/3a/3b/7 主要在 245FEAD248 处裂解，产生的 NT 片段（NT248）可诱导强有力的热凋亡。PoCASP8a/8b 在 202IEKD205 处裂解全长 PoGSDME 和 NT248，从而破坏了 PoGSDME 和 NT248 的生物活性。研究发现了对 PoGSDME 功能至关重要的 9 个残基，其中 F2、L19 和 G85 对质膜转运至关重要。在细菌感染期间，PoGSDME和PoCASP1在比目鱼组织中的表达显著上调，PoGSDME和PoCASP1在比目鱼细胞中的活化伴随着IL-1β和IL-18的加工裂解。这些研究结果揭示了GSDME介导的比目鱼化脓过程的激活和抑制机制，为鱼类化脓过程的调控提供了新的视角。

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引用次数: 0

Corrigendum to "Sargassum horneri extracts stimulate innate immunity, enhance growth performance, and upregulate immune genes in the white shrimp Litopenaeus vannamei" [Fish Shellfish Immunol. 102 (2020) 276-285]. Corrigendum to "Sargassum horneri extracts stimulate innate immunity, enhance growth performance, and upregulate immune genes in the white shrimp Litopenaeus vannamei" [Fish Shellfish Immunol. 102 (2020) 276-285].

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-01 Epub Date: 2024-10-16 DOI: 10.1016/j.fsi.2024.109970

Po-Tsang Lee, Hoang Trieu Quan Tran, Huai-Ting Huang, Fan-Hua Nan, Meng-Chou Lee

引用次数: 0

A novel LPS binding /bactericidal permeability-increasing protein (LBP/BPI) from the scallop Argopecten purpuratus plays an essential role in host resistance to Vibrio infection 扇贝中一种新型 LPS 结合蛋白/杀菌渗透性增加蛋白（LBP/BPI）在宿主抵抗弧菌感染的过程中发挥着重要作用。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-01 DOI: 10.1016/j.fsi.2024.109989

Antonia Jorquera , Camila Montecinos , Yurubí Borregales , Katherine Muñoz-Cerro , Roxana González , Max Santelices , Rodrigo Rojas , Luis Mercado , Felipe Ramírez , Fanny Guzmán , Rodolfo Farlora , Cristian Valenzuela , Katherina Brokordt , Paulina Schmitt

Lipopolysaccharide binding proteins (LBPs) and bactericidal permeability increasing proteins (BPIs) play significant roles in the immune response of vertebrates against bacterial pathogens. These soluble proteins produced by immune cells, specifically interact with and bind to bacterial lipopolysaccharides (LPS), with BPIs also displaying antibacterial activity. In Argopecten purpuratus scallop larvae resistant to Vibrio bivalvicida VPAP30, we identified a significant overexpression of a transcript displaying molecular features of an LBP/BPI protein, both before and after infection. Therefore, in the present work we aimed to understand the role of this novel LBP/BPI, named ApLBP/BPI3, in the scallop resistance to this Vibrio. The ApLBP/BPI3 open reading frame encodes a putative protein of 506 amino acids, with a molecular weight 56.78 kDa. The protein contains a C-terminal domain of 403-amino acid that, after theorical cleavage, displays a soluble form of 44.99 kDa, featuring two BPI/LBP/CETP domains, an apolar binding pocket, a single disulfide bond and a BPI dimerization interface. Phylogenetic analysis reveals high similarity between ApLBP/BPI3 and other mollusk LBP/BPI proteins. Aplbp/bpi3 transcripts were constitutively and highly expressed in hemocytes, gills, mantle, and digestive gland tissues, and were induced following VPAP30 infection in scallop larvae and adult hemocytes. We characterized ApLBP/BPI3 protein using a polyclonal antibody against a synthetic peptide. ApLBP/BPI3 was secreted to the media by infected cultured hemocytes, detected by ELISA. ApLBP/BPI3 was spotted inside non-infected hemocytes, bound to the cell wall of V. bivalvicida after in vitro hemocyte infection, and coating the gills and mantle epithelial barriers before and after an in vivo immune challenge, with stronger detection after VPAP30 injection, detected by immunofluorescence. Infected scallop larvae showed increased ApLBP/BPI3 levels, with slightly higher production in resistant larvae, determined by Western blot. Finally, silencing the Aplbp/bpi3 transcript through RNA interference and and subsequently infecting scallop juveniles with an LD50 of V. bivalvicida resulted in 100 % mortality. Altogether, results demonstrate the essential role of this immune effector in the resistance of A. purpuratus.

脂多糖结合蛋白（LBPs）和杀菌渗透性增加蛋白（BPIs）在脊椎动物对细菌病原体的免疫反应中发挥着重要作用。这些由免疫细胞产生的可溶性蛋白能与细菌脂多糖（LPS）特异性地相互作用并结合，其中 BPIs 还具有抗菌活性。在对双瓣弧菌 VPAP30 具有抗性的紫扇贝幼体中，我们发现在感染前后都有一个转录本显著过表达，该转录本显示了 LBP/BPI 蛋白的分子特征。因此，在本研究中，我们旨在了解这种名为 ApLBP/BPI3 的新型 LBP/BPI 在扇贝对这种弧菌的抗性中的作用。ApLBP/BPI3 开放阅读框编码一个 506 个氨基酸的假定蛋白，分子量为 56.78 kDa。该蛋白含有一个 403 个氨基酸的 C 端结构域，经过理论裂解后，其可溶性形式为 44.99 kDa，具有两个 BPI/LBP/CETP 结构域、一个极性结合袋、一个二硫键和一个 BPI 二聚化界面。系统进化分析表明，ApLBP/BPI3 与其他软体动物的 LBP/BPI 蛋白具有高度相似性。Aplbp/bpi3 转录本在血球、鳃、套膜和消化腺组织中组成型高表达，并在扇贝幼体和成体血球感染 VPAP30 后被诱导表达。我们使用针对合成肽的多克隆抗体鉴定了 ApLBP/BPI3 蛋白。通过酶联免疫吸附法检测，ApLBP/BPI3 被感染的培养血细胞分泌到培养基中。在体内免疫挑战前后，ApLBP/BPI3 在未感染的血细胞内被发现，在体外血细胞感染后与双壳裂头蚴的细胞壁结合，并包被在鳃和套膜上皮屏障上，在注射 VPAP30 后检测更强，用免疫荧光法检测。受感染的扇贝幼体显示 ApLBP/BPI3 水平升高，通过 Western 印迹法测定，抗性幼体的 ApLBP/BPI3 产量略高。最后，通过 RNA 干扰沉默 Aplbp/bpi3 转录本，然后用半数致死剂量的双壳裂头蚴感染扇贝幼体，死亡率为 100%。总之，研究结果表明，这种免疫效应因子在紫扇贝的抗性中起着至关重要的作用。

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引用次数: 0

Evaluation of inhibitory, immunomodulatory, survival, and growth effects of host-derived Weissella confusa on Macrobrachium rosenbergii challenged with Vibrio parahaemolyticus 评估宿主产生的魏氏梭菌对受到副溶血性弧菌挑战的鲭鱼的抑制、免疫调节、存活和生长效应

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-01 DOI: 10.1016/j.fsi.2024.109964

Kristelle Mae C. Tardecilla , Mary Beth B. Maningas

Macrobrachium rosenbergii is a highly valuable prawn species in aquaculture due to its current growing demand in the market. However, various bacterial diseases caused by Vibrio parahaemolyticus have been observed to induce mortality in larval, juvenile, and adult stages of M. rosenbergii. While antibiotics are commonly used to manage these diseases, they contribute to antimicrobial resistance, a global concern with serious environmental and health implications. Consequently, the scientific community is exploring host-derived beneficial microorganisms as functional feed additives, offering eco-friendly alternatives that boost shrimp and prawn immunity. The purpose of this study was to examine the effects of host-derived beneficial microorganisms from the gut of M. rosenbergii, on the inhibition, immunomodulation, survival, and growth of M. rosenbergii when exposed to V. parahaemolyticus. The result of this study showed that Weissella confusa C6, a lactic acid bacterium isolated from the gut of M. rosenbergii, significantly inhibits the growth of V. parahaemolyticus R1 as demonstrated in agar well diffusion assay and co-culture assay. Additionally, it showed immunomodulatory effects, indicated by elevated total hemocyte count (THC) and phenoloxidase (PO) activity during the bacterial challenge test. Furthermore, supplementing the diet with W. confusa led to significant improvements in M. rosenbergii, including increased survival rates, weight gain, specific growth rate, better feed conversion ratio, and reduced density of V. parahaemolyticus in the prawn's gut. Therefore, it can be considered a viable option for use as a feed supplement in prawn farming, enhancing M. rosenbergii's immune system, disease resistance, and overall health.

由于目前市场需求量不断增长，罗氏沼虾（Macrobrachium rosenbergii）在水产养殖业中是一种非常珍贵的对虾品种。然而，由副溶血性弧菌引起的各种细菌性疾病已被观察到会导致对虾幼体、幼鱼和成鱼死亡。虽然抗生素通常用于控制这些疾病，但它们会导致抗菌药耐药性，这是一个全球关注的问题，对环境和健康都有严重影响。因此，科学界正在探索将源自宿主的有益微生物作为功能性饲料添加剂，提供可提高对虾免疫力的生态友好型替代品。本研究的目的是检测从罗氏沼虾肠道中提取的宿主有益微生物对罗氏沼虾暴露于副溶血性弧菌时的抑制、免疫调节、存活和生长的影响。这项研究的结果表明，在琼脂井扩散试验和共培养试验中，从罗氏鹅膏菌肠道中分离出来的乳酸菌魏氏菌 C6 能显著抑制副溶血性弧菌 R1 的生长。此外，它还具有免疫调节作用，在细菌挑战试验中，血细胞总数（THC）和酚氧化酶（PO）活性都有所提高。此外，在日粮中添加 W. confusa 还能显著改善罗氏沼虾的生长状况，包括提高存活率、增重、特定生长率、饲料转化率，并降低对虾肠道中副溶血性大肠杆菌的密度。因此，在对虾养殖中将其用作饲料添加剂，提高对虾的免疫系统、抗病能力和整体健康水平，是一种可行的选择。

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引用次数: 0

Characterization, expressional and evolutionary analysis of five fish-specific CCRs (CCR4La, CCR4Lc, CCR12a1, CCR12a2, and CCR12b) in largemouth bass (Micropterus salmoides) 大口鲈鱼（Micropterus salmoides）中五种鱼类特异性 CCR（CCR4La、CCR4Lc、CCR12a1、CCR12a2 和 CCR12b）的特征、表达和进化分析。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-01 DOI: 10.1016/j.fsi.2024.109984

Xiangyu Pi , Xuan Wei , Mingzhu Pan , Eakapol Wangkahart , Qihuan Zhang , Zisheng Wang , Zhitao Qi

CC chemokine receptors (CCRs), the numbers of the G protein-coupled receptor (GPCR) superfamily, had crucial roles in treating infection, inflammation, and tissue damage by binding to their ligands. In this study, five fish-specific CCRs, namely CCR4La, CCR4Lc, CCR12a1, CCR12a2, and CCR12b, were identified in largemouth bass (Micropterus salmoides). The correction of nomenclatures of these CCRs were confirmed by phylogenetic analysis, structural analysis and genomic synteny analysis. Following 1 × 10⁶ CFU/mL and 1 × 10⁷ CFU/mL Edwardsiella piscicida infection, these five CCRs were significantly induced in spleen of largemouth bass, indicating their important roles in the immune response against bacterial infection. Selection pressure analysis revealed that CCR4La, CCR4Lc, CCR12a1, and CCR12a2 underwent negative selection pressure, whereas CCR12b experienced positive selection pressure. Robust selection site detection methods identified that positive selected sites of CCR4La, CCR4Lc, CCR12a1, and CCR12a2 mainly distributed in their extracellular regions, which involved in ligand binding and pathogen interaction. Similarly, the positive selected sites of CCR12b were also located in its extracellular regions. The accuracy of the pressure selected sites were also validated by molecular docking analysis. The potential ligands for these five CCRs were identified by molecular docking analysis, with finding that CCL3 and CCL5 might be the ligands of largemouth bass CCR4La/Lc, and CCL5, CCL8, CCL7, CCL13 and CCL26 might be that of largemouth bass CCR12a1/a2/b. Our results provided basis for elucidating the functions of chemokine-receptor complex in largemouth bass.

CC 趋化因子受体（CCR）是 G 蛋白偶联受体（GPCR）超家族中的一种，通过与其配体结合，在治疗感染、炎症和组织损伤方面发挥着至关重要的作用。本研究在大口鲈鱼（Micropterus salmoides）中发现了五种鱼类特异性 CCR，即 CCR4La、CCR4Lc、CCR12a1、CCR12a2 和 CCR12b。系统进化分析、结构分析和基因组同源分析证实了这些 CCRs 的命名更正。在感染 1×106 CFU/mL 和 1×107 CFU/mL Edwardsiella piscicida 后，这五种 CCRs 在大口鲈鱼脾脏中被显著诱导，表明它们在抗细菌感染的免疫反应中发挥着重要作用。选择压力分析表明，CCR4La、CCR4Lc、CCR12a1和CCR12a2经历了负选择压力，而CCR12b经历了正选择压力。稳健的选择位点检测方法发现，CCR4La、CCR4Lc、CCR12a1和CCR12a2的正选择位点主要分布在细胞外区域，参与配体结合和病原体相互作用。同样，CCR12b 的阳性选择位点也位于其细胞外区域。分子对接分析也验证了压力选择位点的准确性。通过分子对接分析确定了这五种CCR的潜在配体，发现CCL3和CCL5可能是大口鲈CCR4La/Lc的配体，CCL5、CCL8、CCL7、CCL13和CCL26可能是大口鲈CCR12a1/a2/b的配体。我们的研究结果为阐明大口鲈趋化因子-受体复合物的功能提供了依据。

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引用次数: 0

Recombinant Bacillus subtilis expressing functional peptide and its effect on blunt snout bream (Megalobrama amblycephala) in two state of stress 表达功能肽的重组枯草芽孢杆菌及其对两种应激状态下的钝口鳊鱼（Megalobrama amblycephala）的影响。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-01 DOI: 10.1016/j.fsi.2024.109980

Wei Xiong , Guang-zhen Jiang , Chao-fan He , Hao-kun Hua , Mian-ting Du , Wan-ting Huang , Hui-ting Xu , Mei-ting Zhou , Xi Wang , Hui-xing Guo , Ai-min Wang , Shang-zhi Sun , Wen-bin Liu

This study was conducted to investigate the effects of recombinant Bacillus subtilis CM66-P4' (secreting P4, which related to previous research in this laboratory) on the antioxidant capacity and immune function of blunt snout bream (Megalobrama amblycephala) through in vitro and in vivo experiment. The culture experiment was divided into 3 groups, including control group (CG, with no additional bacteria), original bacteria group (OBG, with 2 × 10⁹ CFU/kg Bacillus subtilis CM66) and recombinant bacteria group (RBG, with 2 × 10⁹ CFU/kg Bacillus subtilis CM66-P4′). After 8 weeks of feeding, a part of the fish were subjected to fishing stress, and the rest were subjected to starvation stress test. Blood samples were collected for the determination of immune and stress-related indexes. The hepatocytes were divided into control group (CG) and experiment group with P4 peptide (LTG and HTG). The cells were collected after starvation treatment and the expression of related genes was detected. The results showed as follows: compared with the CG group, the gene expressions of hepatocytic hsp60 and hsp70 in the LTG and HTG groups were significantly suppressed after 24 h starvation stress (P < 0.05). The content of MDA, the activities of AKP and ALT in OBG group were significantly changed after 30 days starvation (P < 0.05), while the indexes in RBG group had no significant change. The changes of plasma cortisol, malondialdehyde (MDA) and Immunoglobulin M (IgM) in CG and OBG groups were significantly changed at 4 h after fishing stress (P < 0.05), while the indexes in RBG group was not. In conclusion, this study confirmed that Bacillus subtilis CM66-P4′ has great potential in preventing adverse effects of stress on aquatic livestock.

本研究通过体外和体内实验研究了重组枯草芽孢杆菌CM66-P4'（分泌P4，与本实验室以前的研究有关）对钝口鳊抗氧化能力和免疫功能的影响。培养实验分为 3 组，包括对照组（CG，不添加细菌）、原菌组（OBG，添加 2×109 CFU/kg 枯草芽孢杆菌 CM66）和重组菌组（RBG，添加 2×109 CFU/kg 枯草芽孢杆菌 CM66-P4'）。喂养 8 周后，对部分鱼进行捕捞应激试验，其余鱼进行饥饿应激试验。采集血液样本用于测定免疫和应激相关指标。将肝细胞分为对照组（CG）和 P4 肽实验组（LTG 和 HTG）。饥饿处理后收集细胞，检测相关基因的表达。结果表明：与 CG 组相比，LTG 组和 HTG 组肝细胞 hsp60 和 hsp70 的基因表达在饥饿应激 24 h 后明显受到抑制（P < 0.05）。饥饿 30 天后，OBG 组的 MDA 含量、AKP 和 ALT 活性均有明显变化（P < 0.05），而 RBG 组的这些指标无明显变化。CG组和OBG组的血浆皮质醇、丙二醛（MDA）和免疫球蛋白M（IgM）在捕捞应激后4 h发生明显变化（P < 0.05），而RBG组的指标没有明显变化。总之，本研究证实枯草芽孢杆菌 CM66-P4' 在预防应激对水产家畜的不良影响方面具有巨大潜力。

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引用次数: 0

Arginine alleviates LPS-induced leukocytes inflammation and apoptosis via adjusted NODs signaling 精氨酸通过调整 NODs 信号传导减轻 LPS 诱导的白细胞炎症和凋亡。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-01 DOI: 10.1016/j.fsi.2024.109985

Yuan-Xin Qian, Feng-Feng Zhou, Qiang Chen, Fen Dong, Han-Ying Xu, Yu-Long Sun, Ji-Teng Wang, Tao Han

Arginine plays a key role in regulating the immune function of fish. To evaluate the effect of arginine on the immune response of largemouth bass (Micropterus salmoides), the effects of arginine on cell viability, NADPH oxidase activity, respiratory burst activity, and NO production of leukocytes were analyzed both in vitro and in vivo. In this study, we found that arginine could promote the respiratory burst activity of leucocytes both in vivo and in vitro. By incubating leukocytes with the combination of LPS and arginine, we found that arginine supplementation inhibited the expression of inflammatory genes (tumor necrosis factor-alpha, tnfα; interleukin(il) 8 and il10) and apoptotic genes (caspase 3, caspase 8, and caspase 9) induced by LPS, as well as promoted the arginine metabolism. Arginine supplementation significantly induced (cd4-like) cd4 gene expression after LPS challenge. Further studies showed that LPS could significantly increase nucleotide-binding oligomerization domain containing 1 (nod1) gene expression, but decreased the nod2 gene. The arginine supplementation increased nuclear factor kappa-B (NF-κB) protein level. In conclusion, arginine can alleviate LPS-induced inflammatory response and apoptosis as well as induce cd4 gene expression against LPS challenge via adjusting the expression of NODs signaling.

精氨酸在调节鱼类免疫功能方面起着关键作用。为了评估精氨酸对大口鲈鱼（Micropterus salmoides）免疫反应的影响，我们在体外和体内分析了精氨酸对白细胞活力、NADPH 氧化酶活性、呼吸爆发活性和 NO 生成的影响。在这项研究中，我们发现精氨酸在体内和体外都能促进白细胞的呼吸爆发活性。通过将白细胞与 LPS 和精氨酸联合培养，我们发现补充精氨酸可抑制 LPS 诱导的炎症基因（肿瘤坏死因子-α，tnfα；白细胞介素（il）8 和 il10）和凋亡基因（caspase3、caspase8 和 caspase9）的表达，并促进精氨酸的代谢。精氨酸补充能明显诱导 LPS 挑战后（cd4-like）cd4 基因的表达。进一步的研究表明，LPS 能明显增加含核苷酸结合寡聚域 1（nod1）基因的表达，但降低了 nod2 基因的表达。补充精氨酸可提高核因子卡巴-B（NF-κB）蛋白水平。总之，精氨酸能减轻 LPS 诱导的炎症反应和细胞凋亡，并能通过调整 NODs 信号的表达诱导 cd4 基因表达，从而抵御 LPS 的挑战。

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引用次数: 0

Cell death induced by Lepeophtheirus salmonis labial gland protein 3 in salmonid fish leukocytes: A mechanism for disabling host immune responses 鲑鱼唇腺蛋白 3 在鲑鱼白细胞中诱导的细胞死亡：使宿主免疫反应失效的机制。

IF 4.1 2区农林科学 Q1 FISHERIES

Fish & shellfish immunology

Pub Date : 2024-11-01 DOI: 10.1016/j.fsi.2024.109992

Helena Marie Doherty Midtbø , Andreas Borchel , H. Craig Morton , Richard Paley , Sean Monaghan , Gyri Teien Haugland , Aina-Cathrine Øvergård

The salmon louse (Lepeophtheirus salmonis) is an ectoparasite feeding on mucus, skin, and blood of salmonids. On parasitised fish erosions and, at later lice stages, ulcerations appear at the louse feeding site. In susceptible species like Atlantic salmon (Salmo salar) with a limited rejection of lice, only a mild inflammatory response with minor influx of immune cells is seen at these lesions, as the salmon louse secrete proteins that can dampen immune responses. In a previous study, Lepeophtheirus salmonis labial gland protein 3 (LsLGP3) was suggested to dampen cellular responses, and the present study aimed at increasing our understanding of its mode of action. LsLGP3 was found to be secreted on to the host skin, and both in vivo and in vitro experiments were performed to elucidate its function. Histological analysis of the louse attachment site revealed an epidermal and dermal influx of mainly macrophages and granulocytes after 5 days post infestation. The immune cell influx was deeper in the dermis throughout the louse infestation, and LsLGP3 may be involved in dampening this response. Enriched populations of Atlantic salmon B-cells, T-cells, granulocytes, and monocytes were exposed to recombinant LsLGP3 (recLGP3) in vitro, resulting in a significant decrease in cell viability compared to non-exposed controls. An apoptotic cell morphology with “beads-on-a-string” like protrusions was seen in all leukocyte cell fractions after recLGP3 exposure, but not in erythrocytes or keratocytes. A decreased viability was also detected in pink salmon leucocytes, which was not in leucocytes from non-salmonid species. These functional insights suggest that LsLGP3 specifically induces apoptosis of salmonid leukocytes and is likely a key protein secreted by the lice that disables the Atlantic salmon ability to mount an adequate immune response towards the salmon louse. In vivo LsLGP3 knock down studies indicated that the effect is localised primarily at the lice feeding site, without affecting immune cells that are not situated adjacent to the lice-inflicted lesion. The findings from this study could significantly aid in the development of new immune based anti-salmon louse prophylactic measures and treatments.

三文鱼虱（Lepeophtheirus salmonis）是一种外寄生虫，以三文鱼的粘液、皮肤和血液为食。被寄生的鱼身上会出现糜烂，到了后期，虱子的取食部位会出现溃疡。对于像大西洋鲑（Salmo salar）这样对虱子排斥有限的易感鱼种，由于鲑虱分泌的蛋白质会抑制免疫反应，因此在这些病变部位只会出现轻微的炎症反应，免疫细胞也会少量涌入。之前的一项研究表明，三文鱼虱唇腺蛋白 3（LsLGP3）可抑制细胞反应，本研究旨在加深我们对其作用模式的了解。研究发现 LsLGP3 会分泌到宿主皮肤上，并进行了体内和体外实验以阐明其功能。对虱子附着部位的组织学分析表明，虱子侵染 5 天后，表皮和真皮中的巨噬细胞和粒细胞大量涌入。在整个虱子侵扰过程中，免疫细胞涌入真皮层的程度更深，LsLGP3可能参与了抑制这种反应。大西洋鲑的 B 细胞、T 细胞、粒细胞和单核细胞在体外暴露于重组 LsLGP3（recLGP3）后，细胞活力比未暴露的对照组显著下降。暴露于 recLGP3 后，所有白细胞组分中都出现了类似 "串珠状 "突起的细胞凋亡形态，但红细胞或角膜细胞中没有出现这种形态。在粉鲑白细胞中还检测到活力下降，而在非鲑鱼物种的白细胞中则没有发现这种现象。这些功能性研究结果表明，LsLGP3 能特异性地诱导鲑鱼白细胞凋亡，而且很可能是一种由虱子分泌的关键蛋白，它能使大西洋鲑鱼无法对鲑虱做出适当的免疫反应。体内 LsLGP3 基因敲除研究表明，这种影响主要集中在虱子的摄食部位，而不会影响不在虱子感染病灶附近的免疫细胞。这项研究的结果将大大有助于开发基于免疫的新型抗鲑虱预防措施和治疗方法。

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引用次数: 0