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Endoscopic Monitoring of Intestinal Parasites in the European Chub (Squalius cephalus). 欧洲鳙鱼(Squalius cephalus)肠道寄生虫的内镜监测
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-11 DOI: 10.1111/jfd.70103
Aurelie Goutte, Jean-Pierre Ponthus, Léa Lorrain-Soligon

Quantifying intestinal parasite load in freshwater fish requires the development of reliable, minimally invasive methods. We evaluated the use of a very thin and flexible endoscope to detect and quantify acanthocephalan parasites in 25 European chubs (Squalius cephalus). The presence and number of parasites, as estimated using the endoscope, were correlated with dissection-based counts, especially in the anterior part of the intestine accessible via the insertion tube. Accuracy declined in highly infected fish due to overlapping parasites, but endoscopy still provides an accurate evaluation of infection probability and intensity. Thus, this study validates the use of endoscopy as a reliable method for intestinal parasite monitoring in freshwater fish. The deployment of this tool for field-based studies offers promising perspectives for parasitology research.

定量淡水鱼肠道寄生虫负荷需要开发可靠的、微创的方法。我们评估了使用一种非常薄和灵活的内窥镜来检测和量化25只欧洲鲟鱼(头鲨)的棘头寄生虫。使用内窥镜估计的寄生虫的存在和数量与基于解剖的计数相关,特别是在通过插入管可到达的肠道前部。在高度感染的鱼类中,由于寄生虫重叠,准确性下降,但内窥镜检查仍然可以准确评估感染概率和强度。因此,本研究验证了内窥镜作为淡水鱼肠道寄生虫监测的可靠方法。该工具在实地研究中的应用为寄生虫学研究提供了有希望的前景。
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引用次数: 0
Pathologic Description and Genetic Characterisation of Kudoa thunni From Yellowfin Tuna (Thunnus albacares) in the Caribbean Sea. 来自加勒比海黄鳍金枪鱼(Thunnus albacares)的Kudoa thunni的病理描述和遗传特征。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-09 DOI: 10.1111/jfd.70094
Noor-Ul Huda, Magali Duran, Jonnel Edwards, Cynthia Ware, Olando Harvey, Matt J Griffin, David P Marancik

Yellowfin tuna (Thunnus albacares) is a valuable recreational and commercial species in Grenada and the Caribbean region. Myxozoans, specifically the monotypic Kudoidae family within the Order Multivalvulida, are marine parasitic organisms that can negatively impact fish health and marketability. On 8 September 2023, muscle fillets from a yellowfin tuna caught off the coast of Grenada and containing numerous white nodules were submitted for diagnostic evaluation. Gross examination of the muscle showed firm, spherical to ovoid, white polysporous cysts approx. 0.8-1.3 mm in diameter and approx. 10-20 cysts per 2 × 2 cm section of muscle. Cysts were sharply excised and assessed by wet mount cytology revealing a myriad of quadrate myxosporean parasites consistent with the genus Kudoa. Histopathologic examination demonstrated numerous, intracytoplasmic plasmodia with thick hyaline walls containing intact and necrotic myxospores. Parasites present outside of plasmodia were associated with myoliquifaction of the tissue. The case isolate was morphologically consistent with K. thunni and the partial 18S small subunit rDNA (SSU) and 28S large subunit rDNA (LSU) demonstrated high homology (SSU: 99.8%-100%; LSU: 99.5%-100%) to several K. thunni isolates, including those isolated from blackfin tuna (T. atlanticus) harvested near the Caribbean islands of St. Kitts and Nevis. This is the first report of K. thunni infecting yellowfin tuna in the Caribbean region and expands the epidemiologic, pathologic, and molecular record for this parasite.

黄鳍金枪鱼(Thunnus albacares)是格林纳达和加勒比地区有价值的娱乐和商业物种。黏液动物,特别是多阀目的单型Kudoidae家族,是一种海洋寄生生物,对鱼类的健康和适销性产生负面影响。2023年9月8日,在格林纳达海岸捕获的黄鳍金枪鱼肌肉片含有大量白色结节,提交用于诊断评估。肌肉大体检查显示结实,球形至卵球形,约有白色多孢子囊肿。直径0.8-1.3毫米,约;每2 × 2厘米肌肉切片10-20个囊肿。囊肿被急剧切除,并通过湿载细胞学评估,发现无数与Kudoa属一致的方形粘孢子寄生虫。组织病理学检查显示大量的胞浆内疟原虫,具有厚的透明壁,含有完整和坏死的黏液孢子。存在于疟原虫外的寄生虫与组织的肌液化有关。病例分离物在形态上与黑鳍金枪鱼一致,部分18S小亚基rDNA (SSU)和28S大亚基rDNA (LSU)与若干黑鳍金枪鱼分离物具有高度同源性(SSU: 99.8%-100%; LSU: 99.5%-100%),包括从圣基茨和尼维斯加勒比群岛附近收获的黑鳍金枪鱼(T. atlanticus)分离物中分离出来的分离物。这是加勒比地区首次报告黄鳍金枪鱼感染黄鳍金枪鱼,并扩大了该寄生虫的流行病学、病理学和分子记录。
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引用次数: 0
Efficacy of Lactobacillus plantarum CG02 Against Vibrio parahaemolyticus pVPA3-1 Infection in Whiteleg Shrimp (Penaeus vannamei). 植物乳杆菌CG02对凡纳滨对虾副溶血性弧菌pVPA3-1感染的影响
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-09 DOI: 10.1111/jfd.70101
Thao Thi Thu Le, Linh Vu Pham Tran, Hanh My Phan, Thao Phuong Huynh Ngo

Thirty lactic acid bacteria (LAB) isolates were obtained from the digestive tracts of whiteleg shrimp (Penaeus vannamei) and screened for antagonistic activity against Vibrio parahaemolyticus, the causative agent of Acute Hepatopancreatic Necrosis Disease (AHPND). Among these, four isolates demonstrated strong inhibitory effects, producing inhibition zones of 16-20 mm in agar well diffusion assays. These four LAB isolates were reported to cause no shrimp mortality during the feeding trials. Notably, shrimp fed with the isolate CG02 for 4 weeks showed a significantly improved survival (p < 0.05) after the immersion challenge with V. parahaemolyticus strain pVPA3-1. Based on 16S rDNA sequencing, the isolate CG02 was identified as Lactobacillus plantarum. To improve its viability, the isolate CG02 was encapsulated into spray-dried microcapsules. The resulting probiotic powder exhibited high cell viability, with counts of 9 ± 1.45 × 108 CFU g-1 after 4 months at room temperature, and 4.11 ± 0.38 × 108 CFU g-1 after 8 months at 4°C. When incorporated into shrimp feed, the CG02 powder did not cause any shrimp mortality. Moreover, it provided a relative percent survival (RPS) of 34.78% compared to the control group following the immersion challenge with V. parahaemolyticus. These results highlight the potential of spray-dried L. plantarum CG02 as a probiotic feed additive for enhancing shrimp health and supporting sustainable aquaculture practices.

从凡纳滨对虾(Penaeus vannamei)消化道中分离得到30株乳酸菌(LAB),并对其对急性肝胰腺坏死病(AHPND)病原副溶血性弧菌(Vibrio parahemolyticus)的拮抗活性进行了筛选。其中,4株菌株表现出较强的抑制作用,在琼脂孔扩散试验中产生16-20 mm的抑制区。在饲养试验中,这4株乳酸菌均未引起对虾死亡。值得注意的是,用分离物CG02饲喂4周的对虾在室温下4个月的存活率为p 8 CFU g-1,在4℃下8个月的存活率为4.11±0.38 × 108 CFU g-1。将CG02粉掺入对虾饲料中,未引起对虾死亡。此外,与对照组相比,在副溶血性弧菌浸泡攻击后,它的相对存活率(RPS)为34.78%。这些结果突出了喷雾干燥植物乳杆菌CG02作为益生菌饲料添加剂的潜力,可以改善对虾的健康并支持可持续水产养殖。
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引用次数: 0
A Case of Mortality Caused by Bacillus sp. Infection in Farmed, Red-Spotted Masu Trout (Oncorhynchus masou ishikawae) in Japan. 日本养殖红斑马尾鳟鱼(Oncorhynchus masou ishikawae)感染芽孢杆菌致死1例。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-09 DOI: 10.1111/jfd.70098
Yuzo Takada, Tomofumi Kurobe, Ikunari Kiryu, Mari Inada, Kosuke Tomiyama, Eisuke Nakamura, Tetsuya Akutsu, Osamu Kurata

Red-spotted masu trout (Oncorhynchus masou ishikawae; hereinafter, referred as amago trout), is a freshwater salmonid fish species endemic to Japan. From September to October 2023, chronic mortality was observed on an amago trout farm in eastern Japan. To identify the cause of death, bacterial isolation, predominant bacterial species identification, histopathology and experimental challenge test were performed. As a result, bacterial colonies with characteristic features were isolated from all moribund fish. The 16S rRNA and rpoB gene sequences of the isolates showed high similarity with those of Bacillus sp. Identical 16S rRNA and rpoB gene sequences were also detected in moribund fish tissues. The histopathology revealed bacteria in multiple organs, including the spleen, in five of six fish examined. In the experimental challenge test, the fish that were infected with a high dose of bacteria (1.26E+07 cfu per fish) started to die at 3 days post-infection (dpi) and all fish died by 7 dpi. The cumulative mortality rate was lower in the group infected with a lower number of bacteria (1.26E+06 cfu per fish). Our findings identify Bacillus sp. as the disease agent. This is the first report of Bacillus infection in salmonids.

红点马苏鳟鱼(Oncorhynchus masou ishikawae,以下简称马苏鳟鱼)是日本特有的淡水鲑科鱼类。从2023年9月至10月,在日本东部的一个amago鳟鱼养殖场观察到慢性死亡率。通过细菌分离、优势菌种鉴定、组织病理学和实验攻毒试验等方法确定死亡原因。结果,从所有死鱼中分离出具有特征的菌落。分离株的16S rRNA和rpoB基因序列与芽孢杆菌具有较高的相似性。在死鱼组织中也检测到相同的16S rRNA和rpoB基因序列。组织病理学检查显示,在检查的6条鱼中,有5条的多个器官中都有细菌,包括脾脏。实验攻毒试验中,感染高剂量细菌(1.26E+07 cfu /条)的鱼在感染后3天(dpi)开始死亡,7天后全部死亡。感染细菌数量较少的组(每条鱼1.26E+06 cfu)的累积死亡率较低。我们的研究结果确定芽孢杆菌是致病因子。这是关于沙门氏菌感染的第一份报告。
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引用次数: 0
Tissue Tropism and Pathogenesis of LMBV in Largemouth Bass: A Comparative Study of Injection and Immersion Infection Models. 大口黑鲈LMBV的组织趋向性和发病机制:注射和浸泡感染模型的比较研究。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-09 DOI: 10.1111/jfd.70104
Tianmei Hu, Dingzhe Zou, Baozhou Wu, Haiyue Chen, Shuguang Hao, Ying Tian, Wangqian Xu, Yifei Li, Jie Zhou, Run Yang, Jiping Zhang, Yu Liu, Weiwei Zeng

Largemouth bass ranavirus (LMBV) poses a significant threat to the aquaculture industry by causing severe systemic disease and substantial economic losses. This study investigates the tissue distribution, pathological damage, and host response following LMBV-GD1909 infection using two infection models: experimental injection and simulated natural immersion. In the injection model, infected fish exhibited clinical signs, including skin ulcers, haemorrhaging at fin bases, body darkening and organ swelling. Histopathological analysis revealed extensive multi-organ damage, particularly in the liver, spleen, and kidney, characterised by necrosis, haemorrhage, vacuolar degeneration and inflammatory infiltration. Haematological and serum biochemical analyses indicated significant alterations in key parameters, including elevated WBC, RBC and GLU levels, and reduced PLT, TP and LDH levels, reflecting systemic physiological disruption. Viral dynamics analysis demonstrated the pantropic nature of LMBV, with rapid dissemination to multiple tissues post-injection. The liver and spleen were identified as primary target organs, showing the highest viral loads and most severe pathology, indicating a direct correlation between viral load and organ dysfunction. In the immersion model, simulating natural infection, the gills and skin displayed the highest viral loads initially, identifying them as the major entry portals. The gills also served as a key site for viral replication, with a clear gradient of viral spread from external to internal tissues observed over time. This study comprehensively elucidates the tissue tropism, dissemination pathways, and pathological impact of LMBV, providing critical insights for developing targeted prevention and control strategies against LMBV.

大口黑鲈ranv (LMBV)对水产养殖业构成重大威胁,造成严重的全身性疾病和重大的经济损失。本研究采用实验注射和模拟自然浸泡两种感染模型,研究LMBV-GD1909感染后的组织分布、病理损伤和宿主反应。在注射模型中,受感染的鱼表现出临床症状,包括皮肤溃疡、鳍基部出血、身体变黑和器官肿胀。组织病理学分析显示广泛的多器官损害,特别是在肝、脾和肾,以坏死、出血、空泡变性和炎症浸润为特征。血液学和血清生化分析显示,关键参数有显著变化,包括白细胞、红细胞和谷氨酰胺水平升高,PLT、TP和LDH水平降低,反映出全身生理紊乱。病毒动力学分析表明LMBV具有泛性,注射后可迅速传播到多个组织。肝脏和脾脏被确定为主要靶器官,表现出最高的病毒载量和最严重的病理,表明病毒载量与器官功能障碍直接相关。在模拟自然感染的浸泡模型中,鳃和皮肤最初显示出最高的病毒载量,将它们确定为主要的入口。鳃也是病毒复制的关键部位,随着时间的推移,病毒从外部到内部组织的传播有明显的梯度。本研究全面阐明了LMBV的组织向性、传播途径和病理影响,为制定针对性的LMBV防治策略提供了重要见解。
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引用次数: 0
Characterisation of Urocystolithiasis in Farmed Atlantic Salmon (Salmo salar L.): Histopathological Features and Pathophysiological Considerations. 养殖大西洋鲑鱼(Salmo salar L.)尿囊结石的特征:组织病理学特征和病理生理学考虑。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-09 DOI: 10.1111/jfd.70099
Alf Seljenes Dalum, Marta Alarcon, Páll Gretarsson, Anne Katrine Reed, Helene Wisløff, Kari Kaasen McDougall, Marius Takvam

While nephrocalcinosis has received considerable attention in Atlantic salmon aquaculture, mineralisation of the urinary bladder remains largely overlooked. Obstruction of the urinary bladder, well recognised in terrestrials as a potentially fatal condition, is often missed in fish due to its exclusion from routine histological assessments. An increasing number of field cases, colloquially referred to as 'urinary plugs', are now being observed, frequently occurring without or with only slight, nephron mineralisation. This study describes histopathological changes associated with bladder mineral deposits, urocystolithiasis, in farmed Atlantic salmon, including obstruction, urothelial degeneration and intra-renal lesions. The findings highlight urocystolithiasis as an underdiagnosed condition with potential fish health and welfare implications. Functional and histopathological studies targeting the entire urinary system are warranted to elucidate its pathophysiological nature and to support the development of preventative and therapeutic approaches.

虽然肾钙化症在大西洋鲑鱼养殖中受到了相当大的关注,但膀胱矿化在很大程度上仍然被忽视。膀胱梗阻在陆生动物中被认为是一种潜在的致命疾病,但在鱼类中却经常被忽略,因为它被排除在常规的组织学评估之外。现在正在观察到越来越多的实地病例,俗称“尿栓”,通常没有或只有轻微的肾元矿化。本研究描述了养殖大西洋鲑鱼膀胱矿物质沉积、尿囊结石相关的组织病理学变化,包括梗阻、尿路上皮变性和肾内病变。研究结果强调尿囊结石是一种未被诊断的疾病,具有潜在的鱼类健康和福利影响。针对整个泌尿系统的功能和组织病理学研究是必要的,以阐明其病理生理性质,并支持预防和治疗方法的发展。
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引用次数: 0
Epidemiology of Salmonid Rickettsial Septicemia (SRS) in Farmed Salmon: The Role of Sea Lice Infestations in Mortality Risk. 养殖鲑鱼立克次体败血症(SRS)的流行病学:海虱感染在死亡风险中的作用。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-09 DOI: 10.1111/jfd.70097
Benjamín Diethelm-Varela, Nicolhole Atero, Francisca Córdova-Bührle, Enrico L Rezende, Stefan Gelcich, Osvaldo Sandoval, Carlos Navarro, Fernando O Mardones

Piscirickettsia salmonis , the causal agent of salmonid rickettsial septicemia (SRS), is the main pathogen affecting farmed salmonids in Chile. Outbreaks of SRS lead to substantial economic losses for producers. Many determinants related to SRS outcomes are still poorly understood. Here, we conducted a retrospective cohort study from 2014 to 2021 to investigate the epidemiology of SRS at the farm level in southern Chile, employing historical monitoring data. Using time series analysis of weekly SRS mortality risk and sea lice (Caligus rogercresseyi ) infestation levels, we found that SRS mortality risk had a strong seasonal component, with mortalities being significantly higher in the warmer seasons. While Caligus infestation levels have increased significantly over the years, SRS mortality risk has remained constant. Using mixed effects regression models, we identified that a key predictor for both increased weekly SRS mortality risk and higher hazard of reporting the first SRS outbreak of a production cycle was the level of female egg-laying sea lice. We hypothesise that the interaction between sea lice, P. salmonis and rising water temperatures may produce synergistic stress on salmon that accelerates disease progression and prompts overuse of antimicrobials. This calls for an urgent integrated pest management approach in aquaculture practice.

沙门氏菌是导致沙门氏菌立克次体败血症(SRS)的病原体,是影响智利养殖沙门氏菌的主要病原体。SRS的爆发给生产者造成了巨大的经济损失。与SRS结果相关的许多决定因素仍然知之甚少。在这里,我们进行了2014年至2021年的回顾性队列研究,利用历史监测数据调查智利南部农场水平SRS的流行病学。通过对每周SRS死亡率风险和海虱(Caligus rogercresseyi)侵染水平的时间序列分析,我们发现SRS死亡率风险具有很强的季节性成分,在温暖的季节死亡率显著较高。虽然卡利古斯虫的侵袭水平多年来显著增加,但SRS的死亡率风险保持不变。使用混合效应回归模型,我们确定了一个关键的预测因素,即每周SRS死亡风险增加和报告生产周期第一次SRS爆发的风险增加是雌性产卵海虱的水平。我们假设海虱、沙门氏菌和水温上升之间的相互作用可能会对鲑鱼产生协同压力,从而加速疾病进展并促使过度使用抗菌剂。这就要求在水产养殖实践中采取紧急的病虫害综合管理办法。
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引用次数: 0
Establishment of a Hybrid Snakehead Spleen Cell Line for Viral Studies. 用于病毒研究的杂交蛇头脾细胞系的建立。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-06 DOI: 10.1111/jfd.70096
Xuanming Liu, Qing Wang, Xiudong Guo, Yingying Li, Xubing Mo, Jiyuan Yin, Shucheng Zheng, Jian Zhao, Xidong Mu, Yingying Wang

A spleen-derived cell line was established from the spleen of hybrid snakehead (♀Channa argus × ♂Channa maculata) (abbreviated as CAMsp), a species of considerable economic importance in China's freshwater aquaculture, which is severely impacted by the hybrid snakehead rhabdovirus (HSHRV), largemouth bass ranavirus (LMBV), and infectious spleen and kidney necrosis virus (ISKNV). The establishment of a stable in vitro culture system is imperative for the effective isolation, identification, and study of fish viruses. CAMsp, generated through trypsin digestion, had successfully undergone over 80 passages since its initial culture. This cell line exhibited rapid proliferation in Leibovitz's-15 medium (L-15) supplemented with 10% fetal bovine serum at 28°C, achieving monolayer formation within 24 h at a passage ratio of 1:3. Chromosomal analysis of CAMsp at the 60th passage identified a chromosome count of 42; the chromosome number in hybrid snakehead somatic cells is 44, 45, or 46, suggesting chromosomal alterations. Inoculation of CAMsp monolayers with HSHRV, LMBV, and ISKNV resulted in characteristic cytopathic effects (CPE), including cell rounding, aggregation, and eventual detachment. Transmission electron microscopy (TEM) confirmed viral replication and revealed extensive cytopathological changes within the infected cells, demonstrating the susceptibility of the CAMsp cell line to all three viruses. Viral titers, determined by TCID50 assay at 7 days post-infection (dpi), reached 109.25 ± 100.36 TCID50/mL for LMBV, 106·15 ± 100.25 TCID50/mL for ISKNV, and 108·33 ± 100.12 TCID50/mL for HSHRV, indicating efficient viral propagation in this cell line. The CAMsp cell line serves as a valuable model for studying certain fish viruses, virus-host interactions, and disease prevention strategies.

摘要以杂交蛇头(♀Channa argus ×♂Channa maculata)(简称CAMsp)的脾脏为材料,建立了脾脏源细胞系,该物种在中国淡水水产养殖中具有重要的经济价值,严重受到杂交蛇头横纹病病毒(HSHRV)、大口鱼拉纳病毒(LMBV)和传染性脾肾坏死病毒(ISKNV)的影响。建立稳定的体外培养体系是有效分离、鉴定和研究鱼类病毒的必要条件。通过胰蛋白酶消化产生的CAMsp自初始培养以来已成功经历了80多次传代。该细胞系在添加10%胎牛血清的Leibovitz's-15培养基(L-15)中增殖迅速,在28℃条件下以1:3的传代比在24 h内形成单层细胞。对第60代CAMsp进行染色体分析,染色体数为42;杂交蛇头体细胞的染色体数目为44、45或46,表明染色体发生了改变。用HSHRV、LMBV和ISKNV接种CAMsp单层导致特征性的细胞病变效应(CPE),包括细胞变圆、聚集和最终脱离。透射电子显微镜(TEM)证实了病毒复制,并在感染细胞内发现了广泛的细胞病理学变化,证明了CAMsp细胞系对所有三种病毒的易感性。感染后7天(dpi),通过TCID50测定病毒滴度,LMBV为109.25±100.36 TCID50/mL, ISKNV为106·15±100.25 TCID50/mL, HSHRV为108·33±100.12 TCID50/mL,表明病毒在该细胞系中增殖有效。CAMsp细胞系为研究某些鱼类病毒、病毒-宿主相互作用和疾病预防策略提供了有价值的模型。
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引用次数: 0
Persistence and Organ Tropism of Filoviruses in Farmed European Perch (Perca fluviatilis). 养殖欧洲鲈鱼丝状病毒的持久性和器官趋向性。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-06 DOI: 10.1111/jfd.70105
Johanna Perschthaler, Nicole Wildi, Lena K Matthiss, Torsten Seuberlich, Heike Schmidt-Posthaus
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引用次数: 0
Development of an RPA-CRISPR-Cas12a Fluorescence Assay for Rapid and Sensitive Detection of Tilapia Parvovirus (TiPV). RPA-CRISPR-Cas12a荧光法快速灵敏检测罗非鱼细小病毒(TiPV)的建立
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2025-12-02 DOI: 10.1111/jfd.70095
Chu Ma, Yong Zhou, Nan Jiang, Xin Ren, Chen Xu, Na Su, Yuding Fan, Wenzhi Liu

Tilapia parvovirus (TiPV) is an emerging pathogen associated with high mortality rates in farmed tilapia, highlighting the urgent need for rapid and accurate diagnostic tools. In this study, we established an RPA-CRISPR/Cas12a detection system targeting the TiPV NS1 gene. The assay conditions were systematically optimised, including 15-min RPA amplification at 39°C, with reagent concentrations of 200 nM Cas12a, 250 nM crRNA and 200 nM ssDNA reporter. Specificity tests showed no cross-reactivity with other tilapia pathogens (TiLV, S. agalactiae) and other aquatic pathogens (LMBRaV, YcCV, GCRV II, WSSV, CyHV-2, SVCV). Sensitivity evaluation revealed a limit of detection (LoD) of 1.97 × 101copies/μL, which was 100-fold more sensitive than PCR (1.97 × 103copies/μL). Clinical validation with 20 tilapia samples demonstrated a 50% positive detection rate for RPA-CRISPR/Cas12a, 15% higher than PCR (35%). This integrated method combines the advantages of RPA and CRISPR-based signal transduction, offering a field-applicable solution for TiPV monitoring in resource-limited aquaculture environments.

罗非鱼细小病毒(TiPV)是一种新出现的病原体,与养殖罗非鱼的高死亡率有关,突出表明迫切需要快速和准确的诊断工具。本研究建立了针对TiPV NS1基因的RPA-CRISPR/Cas12a检测系统。系统优化检测条件,在39℃条件下,Cas12a试剂浓度为200 nM, crRNA试剂浓度为250 nM, ssDNA报告基因试剂浓度为200 nM, RPA扩增15 min。特异性试验显示,与其他罗非鱼病原体(TiLV、S. agalactiae)和其他水生病原体(LMBRaV、YcCV、GCRV II、WSSV、CyHV-2、SVCV)无交叉反应。检测限为1.97 × 101copies/μL,灵敏度为PCR (1.97 × 103copies/μL)的100倍。20份罗非鱼样本的临床验证表明,RPA-CRISPR/Cas12a的阳性检出率为50%,比PCR(35%)高15%。该综合方法结合了RPA和基于crispr的信号转导的优势,为资源有限的水产养殖环境中TiPV的监测提供了一种适用于现场的解决方案。
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引用次数: 0
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Journal of fish diseases
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