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Magical role of iron nanoparticles for enhancement of thermal efficiency and gene regulation of fish in response to multiple stresses 纳米铁粒子在提高鱼类热效率和基因调控以应对多重压力方面的神奇作用。
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-09 DOI: 10.1016/j.fsi.2024.109949
Neeraj Kumar, Paritosh Kumar, Kotha Sammi Reddy
The present study addresses the challenges of uncontrolled temperature and pollution in aquatic environments, with a focus on fish ability to tolerate high temperature. The investigation aimed to determine the role of iron nanoparticles (Fe-NPs) in enhancing the thermal tolerance of Pangasianodon hypophthalmus exposed to high-temperature stress, arsenic (As), and ammonia (NH3) toxicity. Fe-NPs were synthesized using green approaches, specifically from fish gill. The dietary Fe-NPs were formulated and supplemented at 10, 15, and 20 mg kg⁻1 of feed. Notably, Fe-NPs at 15 mg kg⁻1 diet significantly reduced the critical thermal minimum (CTmin) (14.44 ± 0.21 °C) and the lethal thermal minimum (LTmin) (13.46 ± 0.15 °C), compared to the control and other treatment groups. Conversely, when Fe-NPs at 15 mg kg⁻1 were supplemented with or without exposure to stressors (As + NH3+T), the critical thermal maximum (CTmax) increased to 47.59 ± 0.16 °C, and the lethal thermal maximum (LTmax) increased to 48.60 ± 0.37 °C, both significantly higher than the control and other groups. A strong correlation was observed between LTmin and CTmin (R2 = 0.90) and between CTmax and LTmax (R2 = 0.98). Furthermore, dietary Fe-NPs at 15 mg kg⁻1 significantly upregulated the expression of stress-related genes, including HSP70, iNOS, Caspase-3a, CYP450, MT, cat, sod, gpx, TNFα, IL, TLR, and Ig. The enhanced thermal tolerance (LTmin and LTmax) can be attributed to these gene regulations, suggesting the mechanistic involvement of Fe-NPs in improving thermal resilience. Overall, the findings demonstrate that dietary supplementation with Fe-NPs, particularly at 15 mg kg⁻1, improves thermal tolerance and stress response in P. hypophthalmus by enhancing gene expression and overall thermal efficiency under stressor conditions.
本研究针对水生环境中温度失控和污染所带来的挑战,重点关注鱼类耐高温的能力。该研究旨在确定铁纳米粒子(Fe-NPs)在提高暴露于高温胁迫、砷(As)和氨(NH3)毒性的鳙鱼(Pangasianodon hypophthalmus)的耐热性方面的作用。Fe-NPs采用绿色方法合成,特别是从鱼鳃废料中提取。膳食中的 Fe-NPs 按 10、15 和 20 mg kg-1 的剂量进行配制和给药。值得注意的是,与对照组和其他处理组相比,日粮中 15 毫克/千克的 Fe-NPs 可将临界最低温度(CTmin)显著降至 14.44±0.21 °C,将致死最低温度(LTmin)显著降至 13.46±0.15 °C。相反,当给予 15 mg kg-1 的 Fe-NPs 并暴露或不暴露于应激源(As+NH3+T)时,临界热最高值(CTmax)升至 47.59±0.16°C,致死热最高值(LTmax)升至 48.60±0.37°C,均显著高于对照组和其他处理组。LTmin和CTmin之间(R2 = 0.90)以及CTmax和LTmax之间(R2 = 0.98)具有很强的相关性。此外,15 mg kg-1 的膳食 Fe-NPs 能显著上调多个应激相关基因的表达,包括 HSP70、iNOS、Caspase-3a、CYP450、MT、cat、sod、gpx、TNFα、IL、TLR 和 Ig。热耐受性的增强(LTmin 和 LTmax)可归因于这些基因的调节,这表明 Fe-NPs 参与了提高热适应性的机制。总之,研究结果表明,膳食补充 Fe-NPs(尤其是 15 毫克/千克)可通过提高基因表达和应激条件下的整体热效率,改善鳙鱼的热耐受性和应激反应。
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引用次数: 0
The impact of crowding stress on growth and intestinal integrity in largemouth bass (Micropterus salmoides): Insights into ER stress, autophagy and apoptosis 拥挤压力对大口鲈鱼(Micropterus salmoides)生长和肠道完整性的影响:ER应激、自噬和细胞凋亡的启示
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-09 DOI: 10.1016/j.fsi.2024.109955
Dadong Yan , Lei Gan , Xianghong Dong , Huaimao Tie , Chao Luo , Zhenlu Wang , Haibo Jiang , Jiangfeng Chen , Miao An , Chuanjie Qin , Zhiyuan Lu
Crowding stress is a common abiotic stressor in intensive aquaculture that negatively impacts fish species, leading to growth depression. This study primarily explored the effects of crowding stress on the growth and intestinal integrity of largemouth bass (Micropterus salmoides). A 10-week feeding experiment was conducted with two groups: a control group (0.55 kg/m³) and a crowding stress group (1.10 kg/m³). The results showed that crowding stress significantly impaired fish growth and compromised intestinal integrity. To further elucidate the underlying mechanisms, we investigated the effects of crowding stress on endoplasmic reticulum (ER) stress, autophagy and apoptosis. Our findings revealed an increased proportion of terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL)-positive cells and ER swelling in the intestines of stressed fish, along with a higher number of autophagosomes. Furthermore, there was a significant upregulation in the mRNA expression of genes related to ER stress, autophagy and apoptosis, including GRP78, eIF2α, IRE1, atg3, LC3-2, ulk1α, P62, and caspase-8. In conclusion, crowding stress negatively affects fish growth and intestinal integrity, potentially through the induction of apoptosis and autophagy associated with ER stress-mediated unfolded protein response (UPR). These findings provided critical insights into how intensive aquaculture disrupts intestinal integrity and inhibits growth in fish, offering a valuable reference for future research aimed at enhancing stress resistance in fish under intensive aquaculture conditions.
拥挤应激是集约化水产养殖中常见的非生物应激源,会对鱼类物种产生负面影响,导致生长抑制。本研究主要探讨拥挤应激对大口鲈鱼(Micropterus salmoides)生长和肠道完整性的影响。研究人员进行了为期 10 周的喂食实验,实验分为两组:对照组(0.55 kg/m³)和拥挤压力组(1.10 kg/m³)。结果表明,拥挤应激严重影响了鱼类的生长,并损害了肠道完整性。为了进一步阐明其潜在机制,我们研究了拥挤应激对内质网(ER)应激、自噬和细胞凋亡的影响。我们的研究结果表明,应激鱼肠道中末端脱氧核苷酸转移酶(TdT)dUTP镍末端标记(TUNEL)阳性细胞和ER肿胀的比例增加,自噬体的数量也增加了。此外,与ER应激、自噬和细胞凋亡相关的基因(包括GRP78、eIF2α、IRE1、atg3、LC3-2、ulk1α、P62和caspase-8)的mRNA表达量也明显上调。总之,拥挤应激可能通过诱导与ER应激介导的未折叠蛋白反应(UPR)相关的细胞凋亡和自噬,对鱼类的生长和肠道完整性产生负面影响。这些发现为深入了解集约化水产养殖如何破坏肠道完整性和抑制鱼类生长提供了重要见解,为今后旨在增强集约化水产养殖条件下鱼类抗应激能力的研究提供了有价值的参考。
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引用次数: 0
Akt inhibitors prevent CyHV-2 infection in vitro Akt 抑制剂可预防 CyHV-2 体外感染
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-09 DOI: 10.1016/j.fsi.2024.109940
Yu Song , Ye Zhang , Simin Xiao , Pengfei Li , Liqun Lu , Hao Wang
Cyprinid herpesvirus 2 (CyHV-2) is a double-stranded DNA virus that infects goldfish (Carassius auratus) and crucian carp (C. carassius), resulting in substantial mortality rates and significant epidemiological implications. To gain deeper insights into CyHV-2-host interactions and identify potential therapeutic targets, quantitative proteomics analysis was conducted on CyHV-2-infected Ryukin goldfish fin (RyuF-2) cells. Our findings revealed significant alterations in the expression of proteins associated with the PI3K/Akt signaling pathway, which were up-regulated upon viral infection. Building on these observations, we employed LY294002, a specific inhibitor of PI3K, to investigate its impact on viral replication by inhibiting the PI3K/Akt pathway in GiCF cell line derived from the caudal fin of Carassius auratus gibelio (Bloch). Our results demonstrated the inhibition of both CyHV-2 replication and Akt phosphorylation within this pathway. Quercetin, a plant-derived analogue of LY294002, was further investigated for its anti-CyHV-2 effects in vitro as well as its underlying mechanism. The results suggested that quercetin exhibits antiviral properties against CyHV-2 and may exert its effects through mechanisms similar to those observed with LY294002. Given that aquaculture water serves as a vector for aquaculture viral diseases and the release of chemical compounds can lead to pollution of the aquatic environment, our study shifted focus to crude extracts obtained from plants. We confirmed crude quercetin extract derived from Cuminum anisum has antiviral activity against CyHV-2 in vitro. Therefore, based on our identification of the involvement of PI3K/Akt signaling pathway in CyHV-2 replication, along with verification of its antiviral mechanism using LY294002, we propose natural dietary quercetin as a promising candidate for development into a novel anti-CyHV-2 drug.
鲤科疱疹病毒 2(CyHV-2)是一种双链 DNA 病毒,可感染金鱼(Carassius auratus)和鲫鱼(C. carassius),导致大量死亡和重大流行病学影响。为了深入了解 CyHV-2 与宿主的相互作用并确定潜在的治疗目标,我们对感染 CyHV-2 的琉金金鱼鳍(RyuF-2)细胞进行了定量蛋白质组学分析。我们的研究结果表明,与 PI3K/Akt 信号通路相关的蛋白质表达发生了重大变化,这些蛋白质在病毒感染时上调。基于这些观察结果,我们采用了 PI3K 的特异性抑制剂 LY294002,通过抑制 PI3K/Akt 通路来研究其对病毒复制的影响。我们的研究结果表明,CyHV-2 的复制和 Akt 磷酸化在这一通路中都受到了抑制。槲皮素是 LY294002 的植物类似物,我们进一步研究了它在体外抗 CyHV-2 的作用及其内在机制。结果表明,槲皮素具有抗 CyHV-2 病毒的特性,其作用机制可能与 LY294002 类似。鉴于水产养殖水体是水产养殖病毒性疾病的传播媒介,而化学物质的释放会导致水环境污染,我们的研究重点转向了从植物中提取的粗提取物。我们证实,从孜然中提取的粗槲皮素具有体外抗 CyHV-2 病毒的活性。因此,基于我们对 PI3K/Akt 信号通路参与 CyHV-2 复制的鉴定,以及使用 LY294002 对其抗病毒机制的验证,我们建议将天然膳食槲皮素开发成一种新型抗 CyHV-2 药物。
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引用次数: 0
Machine vision-assisted genomic prediction and genome-wide association of spleen-related traits in large yellow croaker infected with visceral white-nodules disease 感染内脏白结核病的大黄鱼脾脏相关性状的机器视觉辅助基因组预测和全基因组关联。
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-08 DOI: 10.1016/j.fsi.2024.109948
Yulin Bai , Miaosheng Feng , Ji Zhao , Jiaying Wang , Qiaozhen Ke , Zhou Jiang , Pengxin Jiang , Sijing Chen , Longyu Chen , Wei Liu , Tingsen Jiang , Yichen Li , Guopeng Tian , Tao Zhou , Peng Xu
High-resolution and high-throughput genotype-to-phenotype studies in fish are rapidly advancing, driven by innovative technologies that aim to address the challenges of modern breeding models. In recent years, machine vision and deep learning techniques, particularly convolutional neural networks (CNNs), have achieved significant success in image recognition and segmentation. Moreover, qualitative and quantitative analysis of disease resistance has always been a crucial field of research in genetics. This motivation has led us to investigate the potential of large yellow croaker visceral white-nodules disease (VWND) in encoding information on disease resistance for the task of accession classification. In this study, we proposed an image segmentation framework for the feature extraction of the spleen after VWND infection based on machine vision. We utilized deep CNNs and threshold segmentation for automatic feature learning and object segmentation. This approach eliminates subjectivity and enhances work efficiency compared to using hand-crafted features. Additionally, we employed spleen-related traits to conduct genome-wide association analysis (GWAS), which led to the identification of 24 significant SNPs and 10 major quantitative trait loci. The results of function enrichment analysis on candidate genes also indicated potential relationships with immune regulation mechanisms. Furthermore, we explored the use of genomic selection (GS) technology for phenotype prediction of extreme individuals, which further supports the predictability of spleen-related phenotypes for VWND resistance in large yellow croakers. Our findings demonstrate that artificial intelligence (AI)-based phenotyping approaches can deliver state-of-the-art performance for genetics research. We hope this work will provide a paradigm for applying deep learning and machine vision to phenotyping in aquaculture species.
在旨在应对现代育种模式挑战的创新技术的推动下,鱼类的高分辨率和高通量基因型到表型研究正在迅速发展。近年来,机器视觉和深度学习技术,特别是卷积神经网络(CNNs),在图像识别和分割方面取得了巨大成功。此外,抗病性的定性和定量分析一直是遗传学研究的重要领域。基于这一动机,我们研究了大黄鱼内脏白结核病(VWND)在编码抗病性信息方面的潜力,以完成入药分类任务。在这项研究中,我们提出了一种基于机器视觉的图像分割框架,用于提取 VWND 感染后脾脏的特征。我们利用深度 CNN 和阈值分割技术进行自动特征学习和对象分割。与手工创建特征相比,这种方法消除了主观性,提高了工作效率。此外,我们还利用脾脏相关性状进行了全基因组关联分析(GWAS),从而确定了 24 个重要 SNP 和 10 个主要数量性状位点。候选基因的功能富集分析结果也显示了与免疫调节机制的潜在关系。此外,我们还探索了基因组选择(GS)技术在极端个体表型预测中的应用,这进一步支持了大黄鱼抗VWND脾脏相关表型的可预测性。我们的研究结果表明,基于人工智能(AI)的表型分析方法可以为遗传学研究提供最先进的性能。我们希望这项工作能为深度学习和机器视觉在水产养殖物种表型分析中的应用提供一个范例。
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引用次数: 0
Antioxidant anthocyanin synergistic immune enhancer nanopeptide C-I20 remarkably enhances the protective effect of largemouth bass against largemouth bass ranavirus 抗氧化花青素协同免疫增强剂纳米肽 C-I20 可显著增强大口鲈鱼对大口鲈鱼病毒的保护作用。
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-08 DOI: 10.1016/j.fsi.2024.109952
Xingchen Huo , Fengxia Zhao , Chunrong Yang , Jianguo Su
Largemouth bass ranavirus (LMBV) infection results in huge economic losses in largemouth bass (Micropterus salmoides) industry. Nanopeptide C-I20 and anthocyanins have a positive effect on promoting immune responses and antioxidant mechanisms in several aquatic organisms, and are therefore used to inhibit LMBV infection. In this study, we developed an LMBV immersion challenge model using three different viral concentrations (1 × 104 copies/mL, 1 × 105 copies/mL, and 1 × 106 copies/mL) to infect largemouth bass, and LMBV-MCP mRNA expression was detected in infected fish. Following infection, the fish exhibited severe external ulceration, redness swelling, and darkening of the skin. Histopathological examination revealed significant necrosis and inflammation in muscle tissue, epithelial cell shedding in renal tubules, macrophage aggregation centers and cellular vacuolization in spleen and head kidney, and cellular hypertrophy in liver. To mitigate LMBV infection, we explored the protective effects of a combined treatment strategy involving C-I20 and anthocyanin. Overall, the combination of anthocyanin and C-I20 demonstrated the highest protective efficacy, significantly reducing viral loads in muscle, liver, spleen, and head kidney. Moreover, this treatment regimen enhanced antioxidant enzyme activities (T-AOC, TSOD, GSH-Px, CAT) and modulated important immune genes (IL-1, IL-8, TNF-α, IL-10, Mx, and IgM) expression. In conclusion, the synergistic application of anthocyanin and C-I20 demonstrates significant efficacy in mitigating LMBV infection. This research introduces a novel and promising approach to managing infectious diseases in aquaculture settings.
大口鲈鱼病毒(LMBV)感染给大口鲈鱼(Micropterus salmoides)产业造成了巨大的经济损失。纳米肽 C-I20 和花青素对促进多种水生生物的免疫反应和抗氧化机制有积极作用,因此可用于抑制 LMBV 感染。在本研究中,我们利用三种不同的病毒浓度(1×104拷贝/毫升、1×105拷贝/毫升和1×106拷贝/毫升)建立了LMBV浸泡挑战模型,感染大口鲈鱼,并在感染鱼体内检测到LMBV-MCP mRNA的表达。感染后,鱼体出现严重的外部溃疡、红肿和皮肤变黑。组织病理学检查显示肌肉组织明显坏死和发炎,肾小管上皮细胞脱落,脾脏和头肾巨噬细胞聚集中心和细胞空泡化,肝脏细胞肥大。为了减轻 LMBV 感染,我们探讨了 C-I20 和花青素联合治疗策略的保护作用。总体而言,花青素和 C-I20 的组合具有最高的保护效果,能显著降低肌肉、肝脏、脾脏和头部肾脏中的病毒载量。此外,这种治疗方案还能提高抗氧化酶活性(T-AOC、TSOD、GSH-Px、CAT),调节重要免疫基因(IL-1、IL-8、TNF-α、IL-10、Mx 和 IgM)的表达。总之,花青素和 C-I20 的协同应用在减轻 LMBV 感染方面具有显著功效。这项研究为管理水产养殖环境中的传染性疾病提供了一种新颖而有前景的方法。
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引用次数: 0
Mapping the proteomic landscape and anti-inflammatory role of Streptococcus parauberis extracellular vesicles 绘制副猪链球菌细胞外囊泡的蛋白质组图谱和抗炎作用。
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-06 DOI: 10.1016/j.fsi.2024.109945
E.H.T. Thulshan Jayathilaka , Mawallage Kankanamge Hasitha Madhawa Dias , M.S.B.W.T.M. Nipuna Sudaraka Tennakoon , Oh Chulhong , Chamilani Nikapitiya , Hyun-Jin Shin , Mahanama De Zoysa
Bacterial extracellular vesicles (BEVs) are nanoscale membrane-bound structures involved in intercellular communication and transport of bioactive molecules. In this study, we described the proteomic insight and anti-inflammatory activity of Streptococcus parauberis BEVs (SpEVs). Proteomics analysis of SpEVs identified 6209 distinct peptides and 1039 proteins. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated enrichment in pathways related to the biosynthesis of aminoacyl tRNA, amino acids, and secondary metabolites. Based on the predicted protein-protein interactions, we discovered key immunological proteins such as IL12A, IL12B, IL8, CD28, and NF-κB between SpEVs and human proteins. Functionally, SpEVs exhibit strong anti-inflammatory activity in LPS-stimulated Raw 264.7 cells by reducing the production of key inflammatory mediators. These include nitric oxide (NO), reactive oxygen species (ROS), inflammatory cytokines such as TNFα and IL6, as well as inflammation-related proteins like inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). qRT-PCR and immunoblotting results clearly indicate that SpEVs modulate the NF-κB and MAPK pathways to induce anti-inflammatory activity. Furthermore, in vivo experiments with zebrafish larvae demonstrated that SpEVs treatment reduced the NO and ROS production with minimal cell mortality. Finally, we validated the anti-inflammatory activity of SpEVs in vivo by systematically assessing the inhibition of NO production, reduction in ROS generation, prevention of cell death, and modulation of NF-κB and MAPK signaling pathways. In conclusion, SpEVs contain rich in unique proteins that play crucial roles in mediating anti-inflammatory effects.
细菌胞外囊泡(BEVs)是一种纳米级膜结合结构,参与细胞间通讯和生物活性分子的运输。在这项研究中,我们描述了副猪链球菌 BEVs(SpEVs)的蛋白质组学特征和抗炎活性。SpEVs的蛋白质组学分析发现了6209种不同的肽和1039种蛋白质。京都基因与基因组百科全书》(KEGG)分析表明,与氨基酰tRNA、氨基酸和次生代谢物的生物合成有关的途径丰富。根据预测的蛋白-蛋白相互作用,我们发现了 SpEVs 与人类蛋白之间的关键免疫蛋白,如 IL12A、IL12B、IL8、CD28 和 NF-κB。在功能上,SpEVs 通过减少关键炎症介质的产生,在 LPS 刺激的 Raw 264.7 细胞中表现出很强的抗炎活性。这些介质包括一氧化氮(NO)、活性氧(ROS)、炎症细胞因子(如 TNFα 和 IL6)以及炎症相关蛋白(如诱导型一氧化氮合酶(iNOS)和环氧化酶-2(COX-2))。qRT-PCR 和免疫印迹结果清楚地表明,SpEVs 可调节 NF-κB 和 MAPK 通路,从而诱导抗炎活性。此外,用斑马鱼幼体进行的体内实验表明,SpEVs 处理可减少 NO 和 ROS 的产生,且细胞死亡率极低。最后,我们通过系统评估抑制 NO 生成、减少 ROS 生成、防止细胞死亡以及调节 NF-κB 和 MAPK 信号通路,验证了 SpEVs 在体内的抗炎活性。总之,SpEVs 富含独特的蛋白质,在介导抗炎作用方面发挥着关键作用。
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引用次数: 0
The influence of temperature and β-1,3-glucans on the occurrence of white spot syndrome virus and white spot disease in post-larvae of Penaeus vannamei shrimp 温度和β-1,3-葡聚糖对凡纳滨对虾后幼体白斑综合征病毒和白斑病发生的影响
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-05 DOI: 10.1016/j.fsi.2024.109938
Alejandra Valladares , Cristóbal Domínguez-Borbor , Fabrizio Echeverria , Fanny Panchana , Irma Betancourt , Stanislaus Sonnenholzner , Bonny Bayot , Jenny Rodríguez
Penaeus vannamei is the main species of shrimp farmed worldwide, and for Ecuador it is the first non-oil economic sector. However, various pathogens, including white spot syndrome virus (WSSV), threaten the sustainability of shrimp farming. P. vannamei larvae are susceptible to WSSV infection via vertical or horizontal transmission. To decrease the incidence of WSSV in PLs, a bioassay was performed by exposing P. vannamei shrimp (PL25), which were negative for WSSV and white spot disease (WSD), to two protective factors, water temperature (T = 24 °C and T = 31 °C), and immunestimulation using β-1,3-glucans (BG) (with and without BG added to the food), using a crossed two-factor design, for 20 days. The incidence of WSSV and WSD was modeled using generalized linear model (GLM). The strength of the association between the response and explanatory variables was estimated using the odds ratio [Exp (Beta)] and interpreted as the level of risk for the incidence of WSSV or WSD in one of the categories compared to the baseline category in the GLM. Odds ratios were considered significant if their 95 % confidence interval did not include the value 1. The results showed that BG had a significant negative effect on the number of WSSV-positive animals. In addition, a lower probability of WSSV infection was determined by combining 31 °C and BG. In this treatment, the WSD injuries were almost undetectable. Most larvae shrimps affected by WSD exhibited injuries to the antennal gland and connective tissue. In situ hybridization analysis revealed that 29 % of shrimp, initially negative by histology, were positive for WSSV. The virus was detected mainly in the nervous tissue of the head, epithelium, connective tissue of the head appendages, and the oral region surrounding the integumental glands. Apoptosis analysis showed a weak signal in nerve tissue but was more intense in epithelial cells of the head appendages, in association with cuticular damage. According to the findings, WSSV could disseminate among shrimp populations by infecting them through the TG and epithelium of the head appendage, followed by connective tissue and nervous tissue. However, applying BG at 31 °C may serve as a protective measure to reduce WSD injuries and restrict WSSV dissemination.
凡纳米对虾(Penaeus vannamei)是世界上主要的对虾养殖品种,也是厄瓜多尔第一个非石油经济部门。然而,包括白斑综合症病毒(WSSV)在内的各种病原体威胁着对虾养殖的可持续性。凡纳米对虾幼体容易通过垂直或水平传播感染 WSSV。为降低凡纳滨对虾中 WSSV 的发病率,采用双因素交叉设计,将 WSSV 和白斑病(WSD)阴性的凡纳滨对虾(PL25)置于水温(T= 24 °C 和 T= 31 °C)和使用 β-1,3-葡聚糖(BG)(食物中添加和不添加 BG)的免疫刺激两种保护因素下,进行了为期 20 天的生物测定。使用广义线性模型(GLM)对 WSSV 和 WSD 的发病率进行建模。使用几率[Exp (Beta)]估算反应和解释变量之间的关联强度,并将其解释为与广义线性模型中的基线类别相比,其中一个类别的 WSSV 或 WSD 发病率的风险水平。如果比率的 95% 置信区间不包括值 1,则认为比率显著。结果显示,BG 对 WSSV 阳性动物的数量有显著的负面影响。此外,将 31 ºC 和 BG 结合使用可降低 WSSV 感染的概率。在这种处理方法中,几乎检测不到 WSD 损伤。大多数受 WSD 影响的幼虾的触角腺和结缔组织都有损伤。原位杂交分析表明,有 29% 的对虾对 WSSV 呈阳性反应,这些对虾最初在组织学上呈阴性反应。病毒主要在头部神经组织、上皮细胞、头部附肢结缔组织和口腔周围的皮脂腺中检测到。细胞凋亡分析显示,神经组织中的信号较弱,但头部附肢上皮细胞中的信号较强,与角质层损伤有关。研究结果表明,WSSV 可通过头部附肢的 TG 和上皮细胞感染虾群,然后再感染结缔组织和神经组织,从而在虾群中传播。然而,在 31 °C下施用BG可作为一种保护措施,减少WSD的伤害并限制WSSV的传播。
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引用次数: 0
The role of pattern recognition receptors in the innate immune system of Chinese mitten crab (Eriocheir sinensis) 模式识别受体在中华绒螯蟹先天免疫系统中的作用
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-05 DOI: 10.1016/j.fsi.2024.109946
Chenchen Shi , Ta-Hui Lin , Chen Qu
Eriocheir sinensis (Chinese mitten crab) is one of the main economic species in China, which has evolved an extremely sophisticated innate immune system to fend off disease invasions. However, bacterial and viral infections have caused significant financial losses for the E. sinensis aquaculture in recent years. Making well-informed judgments for the control microbial infections would require a thorough understanding and clarification of the intricate innate immune system of E. sinensis. Innate immunity is essential for the host's defense against invasive pathogens. Pattern recognition receptors (PRRs) initially recognize pathogen-associated molecular patterns (PAMPs) and trigger an innate immune response, causing the generation of inflammatory cytokine and promoting the clearance and control of pathogens. In E. sinensis, Toll/Toll-like receptors, lipopolysaccharide and β-1,3-glucan binding proteins, C-type lectins, galactoside-binding lectins, L-type lectins, scavenger receptors, and down syndrome cell adhesion molecules have been identified to be PRRs that are involved in the recognition of bacteria, fungi, and viruses. In this review, we give a comprehensive overview of the literature regarding PRRs' roles in the immunological defenses of E. sinensis, with the aim of providing clues to the mechanisms of innate immunity.
中华绒螯蟹(Eriocheir sinensis,简称中华绒螯蟹)是中国的主要经济物种之一,它已进化出极其复杂的先天免疫系统来抵御疾病入侵。然而,近年来细菌和病毒感染给中华绒螯蟹养殖造成了巨大的经济损失。要想在控制微生物感染方面做出明智的判断,就必须彻底了解和澄清中华鳗错综复杂的先天免疫系统。先天免疫是宿主抵御入侵病原体的重要手段。模式识别受体(PRRs)最初识别病原体相关分子模式(PAMPs)并触发先天性免疫反应,导致炎性细胞因子的产生,促进病原体的清除和控制。在中华鹅耳枥中,Toll/Toll样受体、脂多糖和β-1,3-葡聚糖结合蛋白、C型凝集素、半乳糖苷结合凝集素、L型凝集素、清道夫受体和绒毛综合征细胞粘附分子已被确认为参与识别细菌、真菌和病毒的PRRs。在这篇综述中,我们全面概述了有关 PRRs 在中华鹅膏菌免疫防御中的作用的文献,旨在为先天性免疫机制提供线索。
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引用次数: 0
Microalgal astaxanthin ameliorates cypermethrin-induced necroptosis and inflammation via targeting mitochondrial Ca2+ homeostasis and the ROS-NF-κB-RIPK3/MLKL axis in carp hepatocytes (Cyprinus carpio) 微藻虾青素通过靶向线粒体 Ca2+ 平衡和鲤鱼肝细胞中的 ROS-NF-κB-RIPK3/MLKL 轴,改善氯氰菊酯诱导的坏死和炎症。
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-05 DOI: 10.1016/j.fsi.2024.109944
Shuoyue Li , Yinuo Liu , Di Li , Kaixuan Zhang , Zequn Zhang , Ziwei Zhang , Jingzeng Cai
Cypermethrin is a toxic pesticide that has infiltrated water bodies due to its widespread use. This contamination has led to detrimental effects on the immune organs of aquatic species, including fish. The natural fat-soluble orange-red carotenoid, astaxanthin (MAT), derived from microalgae, possesses anti-inflammatory, antioxidant, and immunomodulatory properties. To elucidate the mechanism of CY induced damage to carp liver cells and assess the potential protective effects of MAT, we established a carp hepatocyte model exposed to CY and/or MAT. Hepatocytes from carp (Cyprinus carpio) were treated with either 8 μM CY or 60 μM MAT for 24 h. Upon exposure CY, a significant increase in reactive oxygen species (ROS) was observed alongside a diminution in the activities of key antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), suggesting an impairment of cellular antioxidant capacity. Subsequently, acridine orange/ethidium bromide (AO/EB) staining and flow cytometry analysis revealed that hepatocytes exposed to CY exhibited a higher incidence of necroptosis, associated with an elevated mitochondrial Ca2+ concentration, which contributed to cellular dysfunction. Furthermore, exposure to CY also activated the ROS-NF-κB-RIPK3/MLKL signaling pathway, increasing the levels of necroptosis-related regulatory factors (RIP1, RIP3, and MLKL) in hepatocytes and the expression of inflammatory genes (IL-6, IFN-γ, IL-4, IL-1β, and TNF-α), which led to immune dysfunction in hepatocytes. The immunotoxic effects induced by CY were mitigated by MAT treatment, suggesting its potential in alleviating the aforementioned changes caused by CY. Overall, the data suggested that MAT therapy could enhance hepatocyte defenses against CY-induced necroptosis and inflammatory responses by regulating mitochondrial Ca2+ homeostasis and inhibiting the ROS-NF-κB-RIPK3/MLKL signaling cascade. This study elucidated the potential benefits of employing MAT to protect farmed fish from agrobiological hazards during CY exposure, underscoring the practical applications of MAT in aquaculture.
氯氰菊酯是一种有毒杀虫剂,由于广泛使用,已经渗入水体。这种污染对包括鱼类在内的水生物种的免疫器官造成了有害影响。从微藻中提取的天然脂溶性橙红色类胡萝卜素虾青素(MAT)具有抗炎、抗氧化和免疫调节的特性。为了阐明 CY 诱导鲤鱼肝细胞损伤的机制并评估 MAT 的潜在保护作用,我们建立了一个暴露于 CY 和/或 MAT 的鲤鱼肝细胞模型。鲤鱼肝细胞经 8 μM CY 或 60 μM MAT 处理 24 小时后,活性氧(ROS)显著增加,主要抗氧化酶、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活性降低,表明细胞抗氧化能力受损。随后,吖啶橙/溴化乙锭(AO/EB)染色和流式细胞术分析表明,暴露于 CY 的肝细胞出现坏死的几率更高,这与线粒体 Ca2+ 浓度升高有关,从而导致细胞功能障碍。此外,暴露于 CY 还激活了 ROS-NF-κB-RIPK3/MLKL 信号通路,增加了肝细胞中坏死相关调节因子(RIP1、RIP3 和 MLKL)的水平和炎症基因(IL-6、IFN-γ、IL-4、IL-1β 和 TNF-α)的表达,从而导致肝细胞免疫功能紊乱。MAT 治疗减轻了 CY 引起的免疫毒性效应,这表明 MAT 有可能减轻 CY 引起的上述变化。总之,这些数据表明,MAT疗法可通过调节线粒体Ca2+平衡和抑制ROS-NF-κB-RIPK3/MLKL信号级联,增强肝细胞对CY诱导的坏死和炎症反应的防御能力。这项研究阐明了在 CY 暴露期间使用 MAT 保护养殖鱼类免受农业生物危害的潜在益处,强调了 MAT 在水产养殖中的实际应用。
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引用次数: 0
Outer membrane vesicles (OMVs) from Tenacibaculum maritimum as a potential vaccine against fish tenacibaculosis 作为预防鱼类天疱疮潜在疫苗的海产天疱疮外膜囊泡 (OMVs)。
IF 4.1 2区 农林科学 Q1 FISHERIES Pub Date : 2024-10-05 DOI: 10.1016/j.fsi.2024.109943
M. Pilar Escribano, Miguel Balado, Beatriz Santos, Alicia E. Toranzo, Manuel L. Lemos, Beatriz Magariños
Outer membrane vesicles (OMVs) have been gained increasing attention in vaccinology due to their ability to induce strong protective humoral and cell-mediated immunity. The Gram-negative bacterium Tenacibaculum maritimum, the causative agent of marine tenacibaculosis, poses a significant challenge to the global aquaculture industry due to its difficult prophylaxis. In previous studies, we demonstrated that OMV production is a key virulence mechanism in T. maritimum. Building on this, the present study aimed to evaluate the efficacy of a natural, encapsulated multi-antigen vaccine made from adjuvant-free, crude T. maritimum OMVs (Tm-OMVs). A vaccination experiment using SP9.1-OMVs was conducted in juvenile turbot (Scophthalmus maximus L.), followed by a T. maritimum bath challenge. Immune responses in the turbot were assessed by measuring anti-Tm antibody levels and analyzing the expression of eight key immune-related genes (il-1β, il-8, il-22, pcna, c3, cd4-1, ifng2, cd8α). The results showed that immunization with SP9.1-OMVs provided significant protection against T. maritimum infection (RPS = 70 %). Vaccinated fish exhibited a dose-dependent increase in anti-Tm antibody titers in blood plasma, along with rapid induction of both innate (il-1β, il-8, il-22, c3) and adaptive (cd4-1, ifng2, cd8α) immune genes as early as 4 h post-bath challenge. These findings offer new insights into the early immune response of turbot following T. maritimum infection and could serve as a foundation for developing novel OMV-based vaccines.
外膜囊泡 (OMV) 能够诱导强大的保护性体液免疫和细胞介导免疫,因此在疫苗学中日益受到关注。革兰氏阴性细菌海洋tenacibaculimum是海洋tenacibaculosis的致病菌,由于其难以预防,给全球水产养殖业带来了巨大挑战。在以前的研究中,我们证明了 OMV 的产生是海洋栉水母的一个关键毒力机制。在此基础上,本研究旨在评估一种由不含佐剂的粗制海产栉水母 OMVs(Tm-OMVs)制成的天然封装多抗原疫苗的功效。使用SP9.1-OMVs对幼年大菱鲆(Scophthalmus maximus L.)进行了疫苗接种实验,随后对其进行了海产褐斑大菱鲆(T. maritimum)浸浴挑战。通过测量抗Tm抗体水平和分析八个关键免疫相关基因(il-1β、il-8、il-22、pcna、c3、cd4-1、ifng2、cd8α)的表达,评估了大菱鲆的免疫反应。结果表明,用SP9.1-OMV免疫可显著保护鱼免受T. maritimum感染(RPS = 70%)。接种疫苗的鱼血浆中的抗 Tm 抗体滴度呈剂量依赖性增加,同时先天性免疫基因(il-1β、il-8、il-22、c3)和适应性免疫基因(cd4-1、ifng2、cd8α)在浴后四小时就开始快速诱导。这些发现为了解大菱鲆感染海褐斑病后的早期免疫反应提供了新的视角,可作为开发基于 OMV 的新型疫苗的基础。
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引用次数: 0
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Fish & shellfish immunology
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