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Morphological, Histological, and Phylogenetic Characterization of Myxobolus branchioepidermis n. sp. (Myxosporea: Myxobolidae) Infecting the Gills and Skin of the Oranda Goldfish (Carassius auratus) 侵染奥兰达金鱼(Carassius auratus)鳃和皮肤的分枝表皮粘虫的形态学、组织学和系统发育特征。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-12-03 DOI: 10.1111/jfd.14040
Hooman Rahmati-Holasoo, Amin Marandi, Hosseinali Ebrahimzadeh Mousavi, Sara Shokrpoor, Fatemeh Arabkhazaeli, Zahra Ziafati Kafi

Myxobolus Bütschli, 1882 is a highly important genus of fish myxozoans, consisting of many species that cause serious diseases in ornamental cyprinid fish species in different geographic areas by infecting a wide range of organs. This study describes a new Myxobolus species infecting the gill lamellae, gill rakers and skin epidermis of goldfish (Carassius auratus) from Mohammadshahr, Iran, based on detailed morphological, histological and molecular analysis. Mature spores were small and spherical in shape, consisting of two symmetrical shell valves in frontal view, measuring 10.3 (9.8–10.9) μm in length and 10.5 (10.3–11.2) μm in width. Two equal-sized relatively pyriform polar capsules were 5.4 (3.9–6.0) μm long and 4.5 (3.7–5.2) μm wide. The polar tubules were coiled with 4–6 turns. A comparative analysis of the morphological and molecular data confirmed that this species is distinct from other identified myxosporeans. Plasmodia in the cutaneous tissue were found in the epidermis, causing extensive hyperplasia of Malpighian cells and mild inflammatory infiltration. In the gill tissue, plasmodia affected only one side of the lamellae, leading to hyperplasia of the lamellar epithelium, edema, and lymphoplasmacytic branchitis. A novel myxozoan species, Myxobolus branchioepidermis n. sp., has been proposed. The study provides a comprehensive analysis of the morphological, histological, and molecular characteristics of this parasite, which infects the gill lamellae and epidermal layer of the skin in oranda goldfish (C. auratus). The identification of this new species adds to the growing body of research on myxozoan parasites and highlights its potential impact on ornamental fish aquaculture.

黏液虫(Myxobolus b tschli, 1882)是鱼类黏液虫的一个重要属,由许多种组成,通过感染广泛的器官,使不同地理区域的观赏鲤科鱼类严重患病。本文对伊朗Mohammadshahr金鱼(Carassius auratus)鳃片、鳃柄和皮肤表皮感染的一种粘液虫进行了详细的形态学、组织学和分子分析。成熟孢子体积小,呈球形,正面由两个对称的壳瓣组成,长10.3 (9.8-10.9)μm,宽10.5 (10.3-11.2)μm。两个大小相等的相对梨形极性胶囊长5.4 (3.9-6.0)μm,宽4.5 (3.7-5.2)μm。极性小管盘绕4-6圈。形态学和分子数据的比较分析证实了该物种与其他已鉴定的粘孢子虫不同。皮肤组织中的疟原虫见于表皮,引起马尔比氏细胞的广泛增生和轻度炎症浸润。在鳃组织中,疟原虫仅影响板层的一侧,导致板层上皮增生、水肿和淋巴浆细胞性支炎。一个新的黏液动物种——分枝黏液表皮黏液(Myxobolus branchioepidermis n. sp.)被提出。本研究全面分析了这种寄生于金金鱼鳃片和表皮层的寄生虫的形态、组织学和分子特征。这一新物种的发现增加了黏液寄生虫的研究,并强调了它对观赏鱼养殖的潜在影响。
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引用次数: 0
The Co-Infection of ISKNV-II and RGNNV Resulting in Mass Mortality of Juvenile Asian Seabass (Lates calcarifer), Zhuhai, Southern China ISKNV-II和RGNNV共同感染导致珠海亚洲海鲈幼鱼大量死亡。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-12-02 DOI: 10.1111/jfd.14052
Qianqian Sun, Shan Ding, Xiangrong Liu, Wenfeng Zhang, Fangzhao Yu, Yuting Fu, Yong Li, Chuanfu Dong

Infectious spleen and kidney necrosis virus (ISKNV) and nervous necrosis virus (NNV) are two common and important causative agents in marine-cultured fish. However, high viral loads of both ISKNV and NNV in the same clinical case is unusual. In this study, a mass mortality event of Asian seabass Lates calcarifer juveniles occurred in Zhuhai, the main Asian seabass cultured area in mainland China. The fish samples were pooled for pathogen identification and both high viral loads of ISKNV and NNV were detected by real-time microfluidic quantitative PCR and conventional PCR. Immunohistochemistry and immunofluorescence showed that strong ISKNV signals were detected in spleen and liver, while strong NNV signals were detected in brain and eye. The tissue homogenates were inoculated into MFF-1 cell and SSN-1 cell, respectively. After several viral passages, both individual ISKNV and NNV were purely isolated from each other, and designated as ASB-ISKNV-23 and ASB-NNV-23, respectively. The whole genome sequences of ASB-ISKNV-23 and ASB-NNV-23 were determined and annotated. The result showed that ASB-ISKNV-23 and ASB-NNV-23 are composed of 112,236 bp and 4441 bp, respectively. Phylogeny analysis showed that ASB-ISKNV-23 belongs to ISKNV-II sub-genotype and ASB-NNV-23 belongs to RGNNV genotype. Collectedly, coinfection of ISKNV-II and RGNNV were firstly documented in mass mortality of Asian seabass in mainland China. Importantly, both individual ISKNV-II and RGNNV were purely isolated using two different permissive cell lines. Our study provides useful information for better understanding the complex pathogenesis regarding the coinfection with ISKNV and NNV in farmed fish.

传染性脾肾坏死病毒(ISKNV)和神经坏死病毒(NNV)是海水养殖鱼类常见的两种重要病原体。然而,在同一临床病例中,ISKNV和NNV的高病毒载量是不寻常的。本研究在中国大陆亚洲海鱼主要养殖区珠海发生了一起亚洲海鱼幼鱼的大规模死亡事件。收集鱼标本进行病原鉴定,采用实时微流控定量PCR和常规PCR检测ISKNV和NNV的高病毒载量。免疫组织化学和免疫荧光显示,大鼠脾脏和肝脏中检测到较强的ISKNV信号,脑和眼中检测到较强的NNV信号。将组织匀浆分别接种于MFF-1细胞和SSN-1细胞。经过几次传代后,ISKNV和NNV个体被完全分离,分别被命名为ASB-ISKNV-23和ASB-NNV-23。测定ASB-ISKNV-23和ASB-NNV-23的全基因组序列并进行注释。结果表明,ASB-ISKNV-23和ASB-NNV-23分别由112,236 bp和4441 bp组成。系统发育分析表明,ASB-ISKNV-23属于ISKNV-II亚基因型,ASB-NNV-23属于RGNNV基因型。综上所述,ISKNV-II和RGNNV的共同感染在中国大陆的亚洲鲈鱼大量死亡中首次被记录下来。重要的是,个体ISKNV-II和RGNNV都是用两种不同的允许细胞系纯粹分离出来的。我们的研究为更好地理解养殖鱼类中ISKNV和NNV合并感染的复杂发病机制提供了有用的信息。
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引用次数: 0
Antimicrobial Susceptibility and Local Epidemiological Cut-Off Values of Vibrio anguillarum Isolated From Fish Farms in Turkey 土耳其养鱼场分离的鳗弧菌的抗菌药物敏感性和当地流行病学临界值。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-11-28 DOI: 10.1111/jfd.14049
Muhammed Duman, Soner Altun, Izzet Burçin Satıcıoğlu, Kübra Balcı, Merve Taşgın, Nisa Bayrak, Christopher Concha, Claudio D. Miranda, Ruben Avendaño-Herrera

Studies on preventing antimicrobial resistance in aquaculture emphasise the need to responsibly and prudently use antimicrobials, selecting those most effective in controlling and/or reducing mortalities caused by vibriosis. In this study, the distribution of the antimicrobial susceptibility of 28 Vibrio anguillarum isolates from Turkish fish farms was determined using the broth microdilution minimum inhibitory concentration (MIC) test according to the Clinical and Laboratory Standards Institute (CLSI) guideline. The epidemiological cut-off (COWT) values of the V. anguillarum isolates of florfenicol (FLO), tetracycline (TET), doxycycline (DOX), oxolinic acid (OXO), enrofloxacin (ENR) and trimethoprim/sulfamethoxazole (SXT) were calculated using the normalised resistance interpretation (NRI) and ECOFFinder methods. Isolates were categorised as belonging to the fully susceptible wild-type (WT) or non-wild-type (NWT) populations. Calculated COWT values (in μg mL−1) were ≤ 1.0 for FLO and DOX, ≤ 0.5 for TET, ≤ 0.016 for ENR, ≤ 0.032 for OXO, and ≤ 4.0 for SXT using the NRI analysis. Percentages of V. anguillarum isolates categorised as belonging to the NWT population were small for the antimicrobials FLO (10.7%) and SXT (0%), whereas they were higher for the antimicrobials OXO (39.3%) and ENR (39.3%). This is the first study to estimate the local COWT values for antibiotics used in the control of V. anguillarum isolates recovered from farmed fish in Turkey.

关于预防水产养殖中抗菌素耐药性的研究强调需要负责任地和谨慎地使用抗菌素,选择那些在控制和/或减少弧菌病造成的死亡率方面最有效的抗菌素。本研究根据临床和实验室标准协会(CLSI)的指导方针,采用肉汤微稀释最低抑菌浓度(MIC)试验测定了土耳其养鱼场分离的28株鳗弧菌的抗菌药物敏感性分布。采用归一化耐药解释(NRI)和ECOFFinder方法计算氟苯尼考(FLO)、四环素(TET)、多西环素(DOX)、氧喹啉酸(OXO)、恩诺沙星(ENR)和甲氧苄啶/磺胺甲恶唑(SXT)分离株的流行病学截止值(COWT)。分离物被分类为属于完全易感野生型(WT)或非野生型(NWT)群体。NRI分析计算出的cot值(单位为μ mL-1): FLO和DOX≤1.0,TET≤0.5,ENR≤0.016,OXO≤0.032,SXT≤4.0。抗微生物药物FLO(10.7%)和SXT(0%)被归类为NWT种群的鳗鲡弧菌分离物比例较小,而抗微生物药物OXO(39.3%)和ENR(39.3%)的比例较高。这是第一项估计用于控制从土耳其养殖鱼类中分离出来的鳗弧菌的抗生素的当地cot值的研究。
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引用次数: 0
Binding of Infectious Hypodermal and Haematopoietic Necrosis Virus-Like Particles to Mannose Receptor Stimulates Antimicrobial Responses in Immune-Related Tissues of Peneaus vannamei 传染性皮下和造血坏死病毒样颗粒与甘露糖受体的结合刺激凡纳滨对虾免疫相关组织的抗菌反应。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-11-28 DOI: 10.1111/jfd.14051
Wattana Weerachatyanukul, Chettupon Pooljun, Charoonroj Chotwiwatthanakun, Pitchanee Jariyapong

Mannose receptor (MR) is a transmembrane protein and a type of pattern-recognition receptor (PRR) that plays a critical role in the immunity of mammals and fish. In this study, we examined the role of MR in binding with infectious hypodermal and haematopoietic necrosis virus-like particle (IHHN-VLP) and the downstream immune pathway that it triggers in the shrimp Peneaus vannamei. Upon IHHN-VLP challenge, transcripts of MR in P. vannamei (PvMR) increased significantly in all examined tissues, particularly those related to shrimp immunity, including hemocyte, hepatopancreas and gill tissues. Specifically, IHHN-VLP bound to the 34-kDa PvMR protein in shrimp-tissue extracts. Immunohistochemistry results of hemocytes showed that PvMR was initially localised on the plasma membrane but later internalised and dispersed throughout the cytoplasm after IHHN-VLP administration. Binding between IHHN-VLP and PvMR also induced significant upregulation of genes for the antimicrobial peptides (AMPs) penaeidin 3 and crustin, presumably to protect the shrimp against the viral infection. However, knocking down PvMR resulted in down-regulation of all immune-related genes examined. Overall, as an immune-related PRR, PvMR serves as a receptor for invading viruses, which then trigger the expression of AMPs. Strategic designs using PvMR could be developed to either block the interaction of native virus with the host cells or provoke its up-regulation to enhance shrimp immunity, which could open up opportunities to fight against IHHNV infection in shrimp.

甘露糖受体(MR)是一种跨膜蛋白和一种模式识别受体(PRR),在哺乳动物和鱼类的免疫中起着至关重要的作用。在这项研究中,我们研究了MR与传染性皮下和造血坏死病毒样颗粒(ihnn - vlp)结合的作用,以及它在凡纳滨对虾(peneus vannamei)中引发的下游免疫途径。在ihnn - vlp攻击后,凡纳滨对虾MR转录本(PvMR)在所有检测组织中显著增加,特别是与对虾免疫相关的组织,包括血细胞、肝胰腺和鳃组织。具体来说,ihnn - vlp与虾组织提取物中34kda的PvMR蛋白结合。血细胞免疫组化结果显示,PvMR最初定位于质膜,但在ihnn - vlp给药后内化并分散到整个细胞质中。ihnn - vlp与PvMR的结合还诱导了抗菌肽(AMPs)、对苯二甲酸酯(penaeidin 3)和壳蛋白(crust tin)基因的显著上调,可能是为了保护虾免受病毒感染。然而,敲除PvMR会导致所有免疫相关基因的下调。总的来说,作为一种免疫相关的PRR, PvMR作为入侵病毒的受体,然后触发amp的表达。利用PvMR的策略设计可以阻断原生病毒与宿主细胞的相互作用或激发其上调,从而增强对虾的免疫力,这可能为对虾抵抗IHHNV感染开辟机会。
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引用次数: 0
Epidemiological Study of Parasites of the Edible Scallop Aequipecten tehuelchus in Patagonia, Southwest Atlantic Ocean 西南大西洋巴塔哥尼亚食用扇贝寄生虫流行病学研究。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-11-28 DOI: 10.1111/jfd.14047
Nuria Vázquez, Carmen Gilardoni, Florencia Cremonte

This study assesses the health status of the edible Tehuelche scallop, Aequipecten tehuelchus, reporting the epidemiological results of the parasites and their infection levels by histopathological analysis and transmission electron microscopy. Environmental factors (site: West and East domains of the San José gulf, seawater temperature) and host traits (size, physiological condition index, gonad development stages) associated with parasite load (prevalence, mean intensity, parasite abundance and total parasite abundance) were tested with generalised linear models (GLMs). Histopathological results revealed Rickettsia-like organisms (RLOs), several protozoans (Trichodina sp., Nematopsis sp. and APXSc apicomplexans) and metazoans (turbellarians and Tumidotheres maculatus pea crabs) as well as inflammatory lesions in form of granulomas and hemocytic infiltration. The prevalence of the parasitofauna did not vary significantly between gulf domains, except the apicomplexan APXSc that was significantly most prevalent in scallops from the East Domain. However, the mean intensity of infection seemed to be determined by the gulf domains, which present distinct hydrographic regimes, where RLO's, Trichodina sp. and T. maculatus were significantly more intense in scallops from the West, opposite with Nematopsis sp. infection. Total parasite abundance was influenced by seasonal variations, it was negatively correlated with warmer temperatures. The physiological condition index of A. tehuelchus seemed not to be affected by the presence or abundance of any of the parasites. Nevertheless, the apicomplexan APXSc would represent a significant risk to the health of A. tehuelchus due to the pathological conditions, and surveillance of this parasite is needed for a sustainable A. tehuelchus fishery.

本研究对食用扇贝(Aequipecten tehuelchus)的健康状况进行了评估,通过组织病理学分析和透射电镜分析报告了寄生虫的流行病学结果和感染水平。采用广义线性模型(GLMs)对环境因子(地点:圣何塞湾西部和东部海域、海水温度)和宿主性状(体型、生理状况指数、性腺发育阶段)与寄生虫负荷(流行率、平均强度、寄生虫丰度和总寄生虫丰度)的相关性进行了测试。组织病理学结果显示立克次体样生物(RLOs),几种原生动物(Trichodina sp., Nematopsis sp.和APXSc顶复合体)和后生动物(turbellarians和Tumidotheres maculatus豌豆蟹)以及以肉芽肿和血细胞浸润形式的炎症病变。除顶复合体APXSc在东域扇贝中最普遍外,各海湾域间寄生动物群的流行率无显著差异。然而,感染的平均强度似乎是由海湾区域决定的,海湾区域呈现出不同的水文环境,其中RLO, Trichodina sp.和T. maculatus在来自西部的扇贝中明显更强烈,与线虫感染相反。寄生虫总丰度受季节变化的影响,与气温升高呈负相关。各寄生体的存在或丰度对东华依蚊的生理状态指数似乎没有影响。尽管如此,由于其病理状况,顶复合体APXSc对中华绒螯蟹的健康构成了重大威胁,对该寄生虫的监测是实现中华绒螯蟹渔业可持续发展的必要条件。
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引用次数: 0
Professor Ronald John Roberts, CCT, FRCVS, FRCPath, FRSE, (b) 1941, (q) Glasgow 1964, Died 3rd August 2024 罗纳德-约翰-罗伯茨教授,CCT,FRCVS,FRCPath,FRSE,(生于 1941 年),(卒于 1964 年)格拉斯哥,2024 年 8 月 3 日逝世。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-11-25 DOI: 10.1111/jfd.14041
Hamish Rodger
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引用次数: 0
Temperature-Related Effects on Disease Susceptibility and Immune Response in Redband Trout (Oncorhynchus mykiss gairdneri) Following Challenge With Flavobacterium columnare 温度对红带鳟鱼(Oncorhynchus mykiss gairdneri)感染黄杆菌(Flavobacterium columnare)后的疾病易感性和免疫反应的影响
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-11-22 DOI: 10.1111/jfd.14046
Joshua P. Egan, Jie Ma, Veronica L. Myrsell, Zhongqi Chen, Jonathan Masingale, Christopher C. Caudill, Timothy Boyle, Julianna Browning, Shawn R. Narum, Kenneth D. Cain, Paul A. Hohenlohe

Heat stress can increase disease risk in fishes by reducing immune function. Interactions between redband trout (Oncorhynchus mykiss gairdneri) and Flavobacterium columnare, a causative agent of columnaris disease, provide an opportunity to investigate the effects of temperature on immune function and disease resistance during periods of thermal stress. We conducted three trials to characterise differences in immune function and mortality between redband trout held at 18°C and 21°C following challenge with F. columnare. In trial 1, cumulative per cent mortality (CPM) was low and not statistically different between 18°C and 21°C. In trials 2 and 2, we administered higher challenge doses and observed increased CPM overall and significantly greater CPM at 21°C than 18°C. Redband trout upregulated il-8, tnf-α, igm and igt following infection by F. columnare, suggesting that all of these genes may be involved in immune responses to F. columnare infection. We found no differences in the strength of the immune responses between fish held at 21°C versus 18°C. This indicated that 21°C did not elicit sufficient thermal stress to impair immune function and that increased CPM at 21°C versus 18°C was due to enhanced F. columnare virulence.

热应激会降低鱼类的免疫功能,从而增加其患病风险。红带鳟(Oncorhynchus mykiss gairdneri)与柱状黄杆菌(柱状黄杆菌病的致病菌)之间的相互作用为研究热应激期间温度对免疫功能和抗病能力的影响提供了机会。我们进行了三项试验,以确定红带鳟在受到柱状杆菌挑战后,在18摄氏度和21摄氏度条件下免疫功能和死亡率的差异。在试验 1 中,累积死亡率(CPM)较低,18°C 和 21°C之间没有统计学差异。在试验 2 和 2 中,我们施用了较高的挑战剂量,观察到累积死亡率总体上升,21°C 时的累积死亡率明显高于 18°C。红带鳟在感染柱孢蛙后,il-8、tnf-α、igm和igt基因上调,这表明所有这些基因可能都参与了对柱孢蛙感染的免疫反应。我们发现,21°C与18°C条件下鱼类的免疫反应强度没有差异。这表明 21 摄氏度不会引起足够的热应力来损害免疫功能,而 21 摄氏度与 18 摄氏度下 CPM 的增加是由于 F. columnare 的毒力增强所致。
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引用次数: 0
Follicular Cell Hyperplasia (Goitre), Adenoma and Adenocarcinoma of the Thyroid Gland in Fourlined Terapon (Pelates quadrilineatus): Clinical and Histopathological Study: 2022–2023 甲状腺滤泡细胞增生(甲状腺肿)、腺瘤和腺癌在四线鼬(Pelates quadrilineatus)中的临床和组织病理学研究:2022-2023:临床和组织病理学研究:2022-2023 年。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-11-22 DOI: 10.1111/jfd.14048
Hooman Rahmati-Holasoo, Sara Shokrpoor, Amin Marandi

Thyroid disorders can manifest in several forms, including follicular cell hyperplasia (goitre), follicular cell adenoma and follicular cell carcinoma. Following the detection of large and gradually increasing tumour-like masses in the gular region of wild fourlined terapons (Pelates quadrilineatus) housed in various tanks in a public aquarium in June 2022, September 2022 and October 2023, several fish displaying clinical signs were transported to the Ornamental Fish Clinic at the Faculty of Veterinary Medicine, University of Tehran. Upon clinical inspection, the fish had prominent tumour-like masses in the gular area, located beneath the operculum. Upon histological examination of the first series, the masses revealed thyroid well-differentiated hyperplastic follicles that contained colloid and were lined by cuboidal epithelial cells. There were no signs of cellular atypia or mitotic figures. The study of the second series also revealed the existence of cellular atypia, but still no mitotic figures. In the examination of the third series, apart from the earlier lesions, there was evidence of neoplastic cell invasion into adjacent structures such as the adipose tissue and subepithelial layer. In addition, there was moderate cellular pleomorphism and a limited number of mitotic figures. Although the termination of water ozonation and the addition of potassium iodide to the feed were effective in treating thyroid hyperplasia, the treatment for thyroid neoplasia was not successful. The present study provides the evidence of clinical and histopathological findings of follicular cell hyperplasia, adenoma and adenocarcinoma of the thyroid gland in fourlined terapon.

甲状腺疾病有多种表现形式,包括滤泡细胞增生症(甲状腺肿)、滤泡细胞腺瘤和滤泡细胞癌。2022 年 6 月、2022 年 9 月和 2023 年 10 月,在一家公共水族馆的不同鱼缸中饲养的野生四线金枪鱼(Pelates quadrilineatus)的鱼鳃部位发现了大块且逐渐增大的肿瘤样肿块,随后,几条出现临床症状的鱼被送往德黑兰大学兽医系观赏鱼诊所。经临床检查,这些鱼的鳃区有明显的肿瘤样肿块,位于厣下方。对第一批鱼进行组织学检查后发现,肿块内有甲状腺分化良好的增生卵泡,卵泡内含有胶质,内衬为立方体上皮细胞。没有细胞不典型性或有丝分裂的迹象。对第二个系列的研究也发现存在细胞不典型性,但仍然没有有丝分裂图形。在第三个系列的检查中,除了早期病变外,有证据表明肿瘤细胞侵入了邻近结构,如脂肪组织和上皮下层。此外,还有中度的细胞多形性和数量有限的有丝分裂。虽然终止臭氧水和在饲料中添加碘化钾对治疗甲状腺增生有效,但对甲状腺肿瘤的治疗并不成功。本研究提供了四线鳖甲状腺滤泡细胞增生、腺瘤和腺癌的临床和组织病理学发现的证据。
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引用次数: 0
Monitoring Fish Bacterial Pathogens of Wild Fish Species From the South China Sea by Applying Next-Generation Sequencing on Gill Tissue 利用鳃组织的下一代测序技术监测南海野生鱼类的鱼类细菌病原体
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-11-22 DOI: 10.1111/jfd.14050
Shlomi Zrihan, Peleg Itay, Yael Kroin, Nadav Davidovich, Natascha Wosnick, Dan Tchernov, Xiu Pei Koh, Stanley C. K. Lau, Danny Morick

The classic epidemiological triangle model of host—environment—pathogen is recently being reshaped into a tetrahedron, with the growing understanding of the importance of the microbiome in this array. The gills, being a gateway into the fish body, bearing an important role in fish homeostasis, host a complex microbiome that reflects the ambient water, while also showing resemblance to gut microbiome. Next-generation sequencing (NGS) and improvements in data analysis tools enable researchers to gather and analyse a lot more data than ever before, take a closer, more detailed look at microbiota, and gain a much better understanding of the biological processes at work in these complex relations. Here, 16S rRNA amplicons of bacterial DNA extracted from the gills of 36 asymptomatic specimens of three wild fish species from the South China Sea (Nemipterus japonicus, Alepes djebaba, and Saurida tumbil) were sequenced using NGS. Data analyses revealed the presence of 20 potentially pathogenic species, including several zoonotic agents. Gill microbiota exhibited host species-specificity, and expressed a significant difference between demersal and pelagic-amphidromous fish. It is suggested that this method be more widely implemented, in order to gain more insight on ocean ecosystems’ health status, as well as fish stocks of commercial importance.

宿主-环境-病原体这一经典的流行病学三角模型最近被重塑为四面体,人们对微生物组在这一阵列中的重要性有了越来越多的认识。鱼鳃是进入鱼体的门户,在鱼类体内平衡中发挥着重要作用,鱼鳃上寄生着复杂的微生物群,这些微生物群反映了周围水域的情况,同时也与肠道微生物群相似。下一代测序技术(NGS)和数据分析工具的改进使研究人员能够收集和分析比以往更多的数据,更仔细、更详细地观察微生物群,并更好地了解这些复杂关系中的生物过程。在这里,研究人员利用 NGS 对从中国南海三种野生鱼类(日本鳉(Nemipterus japonicus)、姬鲷(Alepes djebaba)和金目鲷(Saurida tumbil))的 36 个无症状标本的鳃中提取的细菌 DNA 的 16S rRNA 扩增子进行了测序。数据分析显示存在 20 种潜在致病物种,其中包括几种人畜共患病原体。鳃微生物群具有宿主物种特异性,在底栖鱼类和中上层溯河鱼类之间表现出显著差异。建议更广泛地采用这种方法,以便更深入地了解海洋生态系统的健康状况以及具有重要商业价值的鱼类种群。
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引用次数: 0
Development of Three Recombinase Polymerase Amplification Assays for the Rapid Visual Detection of Spiroplasma eriocheiris 开发三种重组酶聚合酶扩增测定法,用于快速肉眼检测麦氏螺旋体。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-11-21 DOI: 10.1111/jfd.14031
Yunji Xiu, Ruixin Feng, Zongrui Yang, Lirong Jiang, Yiping Lin, Qingyue Xu, Dong Zheng, Shun Zhou

Spiroplasma eriocheiris is a pathogen in the Chinese mitten crab Eriocheir sinensis and is associated with tremor disease. S. eriocheiris infects virtually all artificially bred crustaceans, resulting in significant economic losses to the aquaculture industry in China. Recombinase polymerase amplification (RPA) is considered a promising technology for the rapid and sensitive detection of disease. For the rapid detection of S. eriocheiris, this study established basic RPA, RPA-EXO and RPA-LFD, which were visualised via agarose gel electrophoresis, SYBR Green I, fluorescence signals and test lines on lateral flow dipsticks. Specific primers and probes were designed based on the 16S ribosomal RNA sequence of S. eriocheiris. The optimal primer pairs for the basic RPA, RPA-EXO and RPA-LFD were RPA-F3/R3, RPA-F3/R3 and RPA-LFD-F3/R3, respectively. The optimal temperatures and times of basic RPA, RPA-EXO and RPA-LFD were 37°C for 25 min, 39°C for 30 min and 39°C for 10 min, respectively. The specificity test indicated that in addition to the positive test results for S. eriocheiris, the other five DNA templates tested negative, demonstrating strong specificity. The results of the sensitivity test revealed that the detection limits of RPA, RPA-EXO and RPA-LFD were 5.77 × 10−2, 5.77 × 10−4 and 1.52 × 10−6 ng/μL, respectively, indicating high sensitivity. This study established three methods for the detection of S. eriocheiris that are characterised by a constant temperature, rapid response, high sensitivity and strong specificity.

中华绒螯蟹螺原体(Spiroplasma eriocheiris)是中华绒螯蟹的一种病原体,与震颤病有关。中华绒螯蟹螺旋体几乎感染了所有人工养殖的甲壳类动物,给中国的水产养殖业造成了巨大的经济损失。重组酶聚合酶扩增(RPA)被认为是一种快速灵敏检测疾病的有前途的技术。本研究建立了基本的 RPA、RPA-EXO 和 RPA-LFD,通过琼脂糖凝胶电泳、SYBR 绿 I、荧光信号和侧流浸渍棒上的检测线对其进行可视化。根据 S. eriocheiris 的 16S 核糖体 RNA 序列设计了特定的引物和探针。基本 RPA、RPA-EXO 和 RPA-LFD 的最佳引物对分别为 RPA-F3/R3、RPA-F3/R3 和 RPA-LFD-F3/R3。基本 RPA、RPA-EXO 和 RPA-LFD 的最佳温度和时间分别为 37°C 25 分钟、39°C 30 分钟和 39°C 10 分钟。特异性检测结果表明,除了红喉嗜血杆菌的检测结果为阳性外,其他五种 DNA 模板的检测结果均为阴性,显示了很强的特异性。灵敏度测试结果显示,RPA、RPA-EXO 和 RPA-LFD 的检测限分别为 5.77 × 10-2、5.77 × 10-4 和 1.52 × 10-6 ng/μL,灵敏度较高。本研究建立了三种检测 S. eriocheiris 的方法,它们具有温度恒定、反应迅速、灵敏度高和特异性强的特点。
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Journal of fish diseases
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