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Impact of Polyonchobothrium magnum on health and gut microbial ecology of African catfish (Clarias gariepinus): Insights from morphological, molecular, and microbiological analyses. Polyonchobothrium magnum 对非洲鲶鱼(Clarias gariepinus)健康和肠道微生物生态的影响:形态学、分子学和微生物学分析的启示。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-09-06 DOI: 10.1111/jfd.14013
Alamira Marzouk Fouad, Haitham G Abo-Al-Ela, Eman A Negm, Maha Abdelhaseib, Abdallah Alian, Naser Abdelsater, Rashad E M Said, Fatma A S Anwar, Doaa H Assar, Sara Abdel-Aal Mohamed

Parasites pose significant challenges to aquaculture and fisheries industries. Our study focuses on the Polyonchobothrium magnum and African catfish to address a potential health issue in aquaculture, explore host-parasite interactions that can help develop effective management practices to ensure fish health and industry sustainability. P. magnum was isolated from the stomach of African catfish (Clarias gariepinus) as the primary site of infection, with a prevalence of 10%. Most affected fish were heavily infected (8 out of 10). Infection was confirmed by sequencing the PCR-targeted region of the nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1) gene, along with light and scanning electron microscopes. The parasite had an elongated scolex with deep bothria, a prominent apical disc wider than the scolex itself, and a four-lobed appearance. The scolex contained a central rostellum divided into two semicircles, bearing 26-30 hooks, with an average of 28. The apical disc had large hooks arranged in four quadrants, with 6-8 hooks each, averaging 7 per quadrant. No neck was observed. Phylogenetic analysis of our sequence showed a 100% match with isolates from Guangzhou, China. In infected fish, the anterior kidney showed increased expression levels of nuclear factor kappa B and lysozyme, but decreased levels of in major histocompatibility complex antigen II. Plasma analysis revealed a significant drop in superoxide dismutase, a rise in interleukin-1 beta, and lower IgM levels compared to non-infected controls. Non-infected fish displayed greater gut microbiota diversity, with dominant families including Moraxellaceae, Enterobacteriaceae, Fusobacteriaceae, and Caulobacteraceae, and prevalent genera such as Acinetobacter, Cetobacterium, and Brevundimonas. In contrast, infected fish exhibited very low diversity, with significantly higher proportions of Enterobacteriaceae (45.99%) and Aeromonadaceae (41.79%) compared to non-infected fish, which had 13.76% and 3.64% respectively. Cetobacterium somerae was prevalent in non-infected fish, while infected fish harboured Aeromonas fluvialis, Plesiomonas shigelloides, and Gallaecimonas xiamenensis. Overall, P. magnum disrupted the immune status and gut microbiota of the host, thereby impacting its health.

寄生虫给水产养殖和渔业带来了重大挑战。我们的研究重点是大鳞多刺鲃和非洲鲶鱼,以解决水产养殖中潜在的健康问题,探索宿主与寄生虫之间的相互作用,从而帮助制定有效的管理方法,确保鱼类健康和行业的可持续发展。从非洲鲶鱼(Clarias gariepinus)的胃中分离出了大鳞栉水母,这是主要的感染部位,感染率为 10%。大多数受感染的鱼都是重度感染(10 条中有 8 条)。通过对烟酰胺腺嘌呤二核苷酸脱氢酶亚基 1(ND1)基因的 PCR 靶区进行测序,并使用光学显微镜和扫描电子显微镜对感染情况进行了确认。寄生虫有一个拉长的鞘膜,两侧有深沟,突出的顶端圆盘比鞘膜本身宽,外观呈四裂状。跗节包含一个中央喙,分为两个半圆,带有 26-30 个钩,平均为 28 个。顶端圆盘上的大钩排列成四个象限,每个象限有 6-8 个钩,平均每个象限 7 个。没有观察到颈部。对我们的序列进行的系统发育分析表明,该序列与中国广州的分离物100%吻合。在受感染的鱼体内,前肾显示核因子卡巴B和溶菌酶的表达水平升高,但主要组织相容性复合抗原II的表达水平下降。血浆分析表明,与未感染的对照组相比,超氧化物歧化酶显著下降,白细胞介素-1β升高,IgM水平降低。未感染鱼类的肠道微生物群多样性较高,主要菌科包括莫拉菌科(Moraxellaceae)、肠杆菌科(Enterobacteriaceae)、镰刀菌科(Fusobacteriaceae)和高杆菌科(Caulobacteraceae),主要菌属包括醋杆菌属(Acinetobacter)、鲸杆菌属(Cetobacterium)和布雷文迪单胞菌属(Brevundimonas)。相比之下,感染鱼类的多样性很低,与未感染鱼类相比,肠杆菌科(45.99%)和气单胞菌科(41.79%)的比例明显较高,分别为 13.76% 和 3.64%。在未感染的鱼类中普遍存在球形杆菌,而在感染的鱼类中则普遍存在绒毛单胞菌、石斑单胞菌和厦门加拉单胞菌。总之,褐斑鱼破坏了宿主的免疫状态和肠道微生物群,从而影响了宿主的健康。
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引用次数: 0
Differential expression of the type III secretion system genes in Yersinia ruckeri: Preliminary investigations in different environmental conditions ruckeri 耶尔森氏菌 III 型分泌系统基因的差异表达:不同环境条件下的初步研究
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-09-06 DOI: 10.1111/jfd.14007
Hadis Rahmatelahi, Simon Menanteau-Ledouble, Astrid S. Holzer, Mansour El-Matbouli, Mona Saleh

Type III secretion system (T3SS) is an important virulence system in Gram-negative bacteria. In this investigation, different environmental conditions that regulate the expression of T3SS genes in Yersinia ruckeri were investigated aimed at obtaining a better understanding about its modulation after various environmental challenges. Four isolates of Y. ruckeri CSF007-82, ATCC29473, A7959-11 and YRNC10 were cultivated under the diverse in vitro challenges iron depletion, high salt, low pH and in the presence of fish serum or in the fish cell culture (Chinook Salmon Embryo – CHSE). The transcriptional modulation of the chromosomal genes ysaV, ysaC, ysaJ and prgH of ysa were investigated using quantitative real-time PCR. The expression of prgH, ysaV, ysaC and ysaJ was differentially expressed in all four strains under evaluation. The highest gene expression levels were observed for Y. ruckeri YRNC10 AN after addition of 0.3 M NaCl in Luria Bertani broth. The results obtained from this study provide initial insights into T3SS responses in Y. ruckeri, which pave the way for further studies aimed at expanding our knowledge on the functional roles of the T3SS genes in Y. ruckeri.

III 型分泌系统(T3SS)是革兰氏阴性细菌的一个重要毒力系统。本研究调查了不同环境条件下对 ruckeri 耶尔森菌 T3SS 基因表达的调控,旨在更好地了解其在各种环境挑战下的调控情况。研究人员在缺铁、高盐、低 pH、鱼血清或鱼细胞培养(奇努克鲑鱼胚胎--CHSE)等不同体外挑战条件下培养了四种拉氏耶尔森菌分离株 CSF007-82、ATCC29473、A7959-11 和 YRNC10。利用实时定量 PCR 技术研究了 ysa 的染色体基因 ysaV、ysaC、ysaJ 和 prgH 的转录调控。prgH、ysaV、ysaC和ysaJ的表达在所有四个受测菌株中均存在差异。在 Luria Bertani 肉汤中加入 0.3 M NaCl 后,Y. ruckeri YRNC10 AN 的基因表达水平最高。本研究的结果提供了对 Y. ruckeri 中 T3SS 反应的初步认识,为进一步研究 Y. ruckeri 中 T3SS 基因的功能作用铺平了道路。
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引用次数: 0
Immune response in diseased and healthy common carp exposed to carp edema virus 接触鲤鱼水肿病毒的患病鲤鱼和健康鲤鱼的免疫反应。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-09-06 DOI: 10.1111/jfd.14012
Radek Machat, Lubomir Pojezdal, Jan Gebauer, Radek Tesarik, Jitka Motlova, Miroslava Palikova, Martin Faldyna

The common carp is one of the most economically valuable freshwater fish worldwide and its aquaculture can be severely affected by the koi sleepy disease (KSD)/carp edema virus disease (CEVD). This study explores a natural outbreak of CEVD in a pond containing both clinically healthy and diseased fish of various origins exposed to the virus. We investigated mRNA expression of genes associated with known antiviral immune mechanisms, such as type I interferon signalling and cell-mediated cytotoxicity, and performed a comprehensive protein expression analysis to highlight differences between the two groups in various organs. Significant differences in expression profiles of common carp with and without clinical signs were found to be strongly dependent on the organ from which the sample originated. Components of the complement cascade, including various C3 proteins, exhibited upregulation only in less affected organs, specifically the head kidney and spleen. Other complement proteins such as B/C2 and C9 showed upregulation in the kidney, spleen, and gills but not in the skin. Conversely, lysozymes C and G, were observed to be upregulated in the most affected organs of the skin and gills. This study submits the first description of the immune system related proteome using a mass spectrometry on the samples isolated from fish infected with CEV. It also offers a unique comparison of immune reaction of CEV infected and healthy fish under an infectious pressure.

鲤鱼是全球最具经济价值的淡水鱼之一,其水产养殖会受到锦鲤昏睡病(KSD)/鲤鱼水肿病病毒病(CEVD)的严重影响。本研究探讨了在一个池塘中自然爆发的 CEVD,该池塘中既有临床健康的鱼,也有接触过病毒的不同来源的病鱼。我们调查了与已知的抗病毒免疫机制(如 I 型干扰素信号传导和细胞介导的细胞毒性)相关的基因的 mRNA 表达,并进行了全面的蛋白质表达分析,以突显两组鱼在不同器官中的差异。研究发现,有临床症状和无临床症状的鲤鱼在表达谱上的显著差异与样本来源器官有很大关系。补体级联的组成成分,包括各种 C3 蛋白,只有在受影响较小的器官,特别是头部肾脏和脾脏,才会出现上调。其他补体蛋白(如 B/C2 和 C9)在肾脏、脾脏和鳃中上调,但在皮肤中没有上调。相反,在受影响最严重的皮肤和鳃等器官中,溶菌酶 C 和 G 则出现上调。本研究首次使用质谱法描述了从感染 CEV 的鱼体内分离的样本中与免疫系统相关的蛋白质组。该研究还对感染 CEV 的鱼和健康鱼在感染压力下的免疫反应进行了独特的比较。
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引用次数: 0
Immune response and protection efficacy of formalin-killed vaccines against Streptococcus iniae in four-finger threadfin Eleutheronema tetradactylum 福尔马林杀灭疫苗对四指线鲷(Eleutheronema tetradactylum)猪链球菌的免疫反应和保护效力。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-08-29 DOI: 10.1111/jfd.14009
Yin-Ze Shi, Andre Giovanni, Li-Wu Cheng, Wen-Rou Huang, Pei-Chi Wang, Shih-Chu Chen

Four-finger threadfin, Eleutheronema tetradactylum farming in southern Taiwan has been facing disease problems caused by Streptococcus iniae since 2018. The development of a vaccine against infectious S. iniae in the cultured threadfin industry is necessary. Thus, this study aimed to examine the efficacy of threadfin immunized formalin-killed cells (FKC) from S. iniae GSI-111 for 42 days post-vaccination (dpv) using two doses of FKC alone (a booster at 14 dpv) as group A, and FKC mixed with ISA763A adjuvant using a single dose as group B or double doses as group C. Immunoglobulin (Ig)-M was purified from threadfin, and rabbit anti-threadfin IgM polyclonal antibodies were used to detect antibody level in immunized fish; the vaccinated group A displayed higher levels at 3 dpv and all vaccinated treatments demonstrated high antibody levels between 14 and 42 dpv. All vaccine groups showed significantly higher values of lysozyme activity at 42 dpv compared with the control group; the vaccinated A group peaked at 14 dpv. The expression profiles of pro-inflammatory and immune-related genes, TNF-α, IL-12A, and C2 were upregulated at 3 dpv, while CD8A and chemokine receptor CXCR4 were upregulated at 42 dpv. Finally, the threadfins were challenged with S. iniae at 42 dpv. The average relative percent survival was 96% for vaccination A and B treatments, and 100% for vaccination C treatment. In summary, this study demonstrated that FKC vaccines whether formulated with an adjuvant could stimulate immune response and effective protect threadfins against S. iniae infection.

自 2018 年以来,台湾南部的四指线鱼养殖业一直面临着由链球菌引起的疾病问题。在养殖螺纹鱼的行业中,有必要开发针对传染性伊氏链球菌的疫苗。因此,本研究旨在研究线鱼免疫福尔马林杀灭细胞(FKC)从 S. iniae GSI-111 疫苗接种后 42 天(dpv)的疗效,以两剂 FKC 单剂(14 dpv 时加强剂)作为 A 组,以 FKC 与 ISA763A 佐剂混合单剂作为 B 组或双剂作为 C 组。从螺纹鱼中纯化出免疫球蛋白(Ig)-M,并使用兔抗螺纹鱼 IgM 多克隆抗体检测免疫鱼的抗体水平;接种疫苗的 A 组在 3 dpv 时显示出较高的抗体水平,所有接种处理在 14 至 42 dpv 之间都显示出较高的抗体水平。所有疫苗组在 42 dpv 时的溶菌酶活性值都明显高于对照组;接种 A 组的溶菌酶活性在 14 dpv 达到峰值。促炎和免疫相关基因、TNF-α、IL-12A和C2的表达谱在3 dpv时上调,而CD8A和趋化因子受体CXCR4在42 dpv时上调。最后,线虫在 42 dpv 时受到猪链球菌的挑战。疫苗 A 和 B 处理的平均相对存活率为 96%,疫苗 C 处理的平均相对存活率为 100%。总之,本研究表明,FKC 疫苗无论是否添加佐剂,都能刺激免疫反应,有效保护螺纹鱼免受猪链球菌感染。
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引用次数: 0
Exploring the antiparasitic and antimicrobial potential of Schinus terebinthifolius Raddi essential oil against fish and shrimp pathogens 探索藜芦精油对鱼虾病原体的抗寄生虫和抗菌潜力。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-08-20 DOI: 10.1111/jfd.14010
Jucimauro de Araújo Pereira Junior, Gracienhe Gomes dos Santos, Domickson Silva Costa, Graziella Vivine Gonçalves de Matos Silva, Paola Capistrano dos Santos, Gabriele Bueno dos Anjos, Luiz Felipe Kretzer de Moraes, Claudia Andrea Lima Cardoso, Arlene Sobrinho Ventura, Natalino da Costa Sousa, Mauricio Laterça Martins

This study aimed to perform in vitro antiparasitic and antimicrobial tests with the essential oil (EO) of Schinus terebinthifolius against of fish and shrimp. The chemical composition of the EO of S. terebinthifolius was determined by gas chromatography. For the antiparasitic test, the protozoan Epistylis sp. obtained from parasitized Oreochromis niloticus was used, and exposed to different concentrations of EO (2%, 1%, 0.5%, 0.25%), and control with 1% grain alcohol. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) test with EO of S. terebinthifolius evaluated the antimicrobial potential, with serial dilutions starting at 2% and control with 1% grain alcohol, using the strains of Aeromonas hydrophila (2.2 × 108 CFU mL−1), Edwardsiella tarda, Vibrio parahaemolyticus, V. harveyi, and V. alginolyticus (2.0 × 108 CFU mL−1). Chemical analysis revealed that the major EO compounds of S. terebinthifolius were δ-3-Carene (56.00%) and α-Pinene (16.89%). In the antiparasitic test, the concentration of 2% EO showed 100% efficacy against Epistylis sp. within 5 min. In the antimicrobial tests, the concentration of 2% EO was effective against all bacteria tested. The EO of S. terebinthifolius demonstrated antiparasitic and antimicrobial activity at a concentration of 2%, standing out as an alternative to conventional antibiotics.

本研究的目的是利用蛇床子精油(EO)对鱼类和虾类进行体外抗寄生虫和抗菌试验。研究采用气相色谱法测定了风铃草精油的化学成分。在抗寄生虫试验中,使用了寄生于黑线鲈的原生动物 Epistylis sp.,将其置于不同浓度的环氧乙烷(2%、1%、0.5%、0.25%)中,并用 1%的谷物酒精进行对照。以 2%的浓度为起点,用 1%的谷物酒精作为对照,对嗜水气单胞菌(Aeromonas hydrophila)菌株进行最小抑菌浓度(MIC)和最小杀菌浓度(MBC)测试,评估其抗菌潜力。2 × 108 CFU mL-1)、Edwardsiella tarda、副溶血性弧菌、V. harveyi 和 V. alginolyticus(2.0 × 108 CFU mL-1)。化学分析显示,风信子的主要环氧乙烷化合物是δ-3-蒈烯(56.00%)和α-蒎烯(16.89%)。在抗寄生虫试验中,浓度为 2% 的环氧乙烷在 5 分钟内对 Epistylis sp.在抗菌测试中,浓度为 2% 的环氧乙烷对所有测试细菌都有效。2%浓度的风信子环氧乙烷具有抗寄生虫和抗菌活性,可作为传统抗生素的替代品。
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引用次数: 0
Assessment of dietary yeast-based additives for cultured catfish and tilapia health 评估膳食酵母添加剂对养殖鲶鱼和罗非鱼健康的影响。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-08-19 DOI: 10.1111/jfd.14008
Guillaume Cacot, D. Allen Davis, Benjamin R. LaFrentz, Mark R. Liles, Ian A. E. Butts, Craig A. Shoemaker, Benjamin H. Beck, Mark Farmer, Timothy J. Bruce

Channel catfish (Ictalurus punctatus) and Nile tilapia (Oreochromis niloticus) are two aquaculture species of great importance. Intensive production is often hindered by poor growth performance and disease mortality. The aim of this study was to evaluate the potential of a commercial fermented yeast product, DVAQUA, on channel catfish and Nile tilapia growth performance metrics and disease resistance. Channel catfish and Nile tilapia were fed practical diets supplemented with 0%, 0.1% or 0.4% of DVAQUA over approximately 2-month feeding periods in recirculation aquaculture systems. To assess the potential of the postbiotic against common aquaculture pathogens, juvenile catfish were subsequently challenged by immersion with Edwardsiella ictaluri S97-773 or virulent Aeromonas hydrophila ML09-119. Nile tilapia juveniles were challenged by injection with Streptococcus iniae ARS-98-60. Serum lysozyme activity, blood chemistry and growth metrics were measured at the end of the feeding period, but no differences were observed across the different metrics, except for survival. For the pathogen challenges, there were no differences in endpoint mortality for channel catfish with either pathogen (p > .05). In contrast, Nile tilapia survivability to S. iniae infection increased proportionally to the inclusion of DVAQUA (p = .005). Changes to sera lysozyme activity were also noted in the tilapia trial, with a reduction of activity in the fish fed the 0.4% DVAQUA diet compared to the control diet (p = .031). Expression profiles of proinflammatory genes and antibodies were also found to be modulated in channel catfish fed the postbiotic, indicating some degree of protective response. These results suggest that this postbiotic may be beneficial in protecting Nile tilapia against S. iniae infection by influencing immune parameters and additional research is needed to evaluate the potential of this DVAQUA for improving catfish health and disease control.

斑点叉尾鮰(Ictalurus punctatus)和尼罗罗非鱼(Oreochromis niloticus)是两种非常重要的水产养殖品种。集约化生产往往受到生长性能差和疾病死亡率高的阻碍。本研究的目的是评估商业发酵酵母产品 DVAQUA 对鲶鱼和尼罗罗非鱼生长性能指标和抗病性的潜在影响。在循环水养殖系统中,用添加了 0%、0.1% 或 0.4% DVAQUA 的实用日粮喂养河鲶和尼罗罗非鱼,喂养期约为 2 个月。为了评估后生菌素对抗常见水产养殖病原体的潜力,鲶鱼幼鱼随后被浸泡在 Edwardsiella ictaluri S97-773 或带毒的嗜水气单胞菌 ML09-119 中。尼罗罗非鱼幼鱼则通过注射猪链球菌 ARS-98-60 接受挑战。饲养期结束时测量血清溶菌酶活性、血液生化指标和生长指标,但除存活率外,不同指标之间未观察到差异。在病原体挑战中,两种病原体对渠道鲶的终点死亡率都没有影响(p > .05)。与此相反,尼罗罗非鱼感染 S. iniae 的存活率随 DVAQUA 的添加而成正比增加(p = .005)。在罗非鱼试验中还注意到血清溶菌酶活性的变化,与对照组相比,饲喂 0.4% DVAQUA 日粮的鱼的溶菌酶活性降低了(p = .031)。研究还发现,饲喂后生物质的沟鲶的促炎基因和抗体的表达谱也受到了调节,这表明其具有一定程度的保护性反应。这些结果表明,这种后生物素可通过影响免疫参数来保护尼罗罗非鱼免受 S. iniae 感染,因此需要进行更多的研究来评估这种 DVAQUA 改善鲶鱼健康和疾病控制的潜力。
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引用次数: 0
Mortality events associated with microsporidian, Spraguea sp., in White Trevally culture in Japan 日本白鲹养殖中与小孢子虫 Spraguea sp.有关的死亡事件。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-08-13 DOI: 10.1111/jfd.14006
Tomofumi Kurobe, Ikunari Kiryu

In 2021, White Trevally or Striped Jack cultured in the western part of Japan exhibited mild, but chronic mortalities from late September through early October. The cumulative mortality rate was approximately 0.02% per a net pen containing approximately 50,000 fish. Although the cumulative mortality rate was not high, most of the fish in net pens showed characteristic gross signs and an abnormal swimming behaviour. The body of diseased fish became pale and the yellow lines on the lateral sides of fish body became darken. In addition, silver lines along the dorsal fin became apparent. Loss of schooling behaviour was noted during the mortality event. In addition, affected fish became lethargic and failed to swim against current, or frequently stopped swimming and sank to the bottom of net pens after feeding. The goal of this study was to identify the cause of the mortality event. To achieve the goal, we used histopathology and metatranscriptome analysis. Histopathological examination revealed that xenoma of microsporidian were frequently observed in the nerve axon in the brain and spinal cord. Spores observed in the sections were stained with a fluorescent dye, Uvitex 2B, indicating those spores are microsporidian. The data from metatranscriptome analysis indicated that the microsporidian is Spraguea sp. The microsporidian was frequently detected from diseased fish with similar symptoms collected in the same region, suggesting that the microsporidian was highly associated with abnormal swimming behaviour of fish.

2021 年,在日本西部养殖的白鲹或条纹鲹从 9 月下旬到 10 月上旬出现了轻微但长期的死亡。每个网箱约有 50,000 条鱼,累计死亡率约为 0.02%。虽然累计死亡率不高,但网箱中的大多数鱼都出现了特征性的体征和异常游动行为。病鱼的身体变得苍白,身体侧面的黄线变黑。此外,背鳍上的银线也变得明显。病鱼在死亡过程中失去了游动能力。此外,受影响的鱼变得昏昏欲睡,不能逆流游动,或在摄食后经常停止游动并沉入网箱底部。本研究的目的是确定死亡事件的原因。为了实现这一目标,我们采用了组织病理学和转录组分析方法。组织病理学检查显示,在大脑和脊髓的神经轴突中经常观察到小孢子虫的异形瘤。切片中观察到的孢子经荧光染料 Uvitex 2B 染色,表明这些孢子为小孢子虫。元转录组分析的数据显示,微孢子虫是 Spraguea sp.。在同一地区采集的具有类似症状的病鱼身上经常检测到微孢子虫,这表明微孢子虫与鱼的异常游泳行为高度相关。
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引用次数: 0
Diversity in clinical isolates of Ictalurid herpesvirus 1 (IcHV1) from U.S. farm-raised catfish and virulence assessment in channel and channel × blue catfish hybrids 来自美国养殖鲶鱼的伊卡疱疹病毒 1 (IcHV1) 临床分离株的多样性以及对河豚和河豚 × 蓝鲶杂交种的毒力评估。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-08-11 DOI: 10.1111/jfd.14005
Arun Venugopalan, Danielle White, Adrián López-Porras, Lorelei Ford, Cynthia Ware, Marsha A. Lewis, James M. Steadman, Lester H. Khoo, Bradley Richardson, Charles M. Walker, Todd S. Byars, David J. Wise, Matt J. Griffin, Larry A. Hanson

Ictalurid herpesvirus 1 (IcHV1) is the most significant viral agent in U.S. catfish aquaculture. Little is known regarding the genetic stability and antigenic variability of IcHV1. Herein, the genetic and antigenic diversity of IcHV1 field isolates was assessed by restriction fragment length polymorphism (RFLP) analysis and serum neutralization assays. RFLP analysis identified two distinct genotypes (IcHV1A and IcHV1B), both discrete from blue catfish alloherpesvirus (BCAHV). Neutralization assays with anti-IcHV1 monoclonal antibody Mab-95 indicate shared antigenic determinants for IcHV1A and IcHV1B that are absent from BCAHV, which Mab-95 did not neutralize. Virulence assessments with representative isolates demonstrate significant differences between isolates within RFLP groups and pooled RFLP group data suggest IcHV1B (pooled survival [mean ± SE]: 58.3% ± 2.6) may be more virulent than IcHV1A (survival: 68.6% ± 2.4). Rechallenges with representative IcHV1A and IcHV1B isolates indicate a cross-protective effect, with fish surviving initial exposure to IcHV1A or IcHV1B showing robust protection when subsequently re-exposed to IcHV1A or IcHV1B. This work demonstrated significant differences in virulence between case isolates, identifying two discrete IcHV1 lineages, distinct from BCAHV, with similar virulence in channel and channel × blue catfish hybrids and a cross-protective effect in catfish surviving exposure to either lineage.

伊科疱疹病毒 1(IcHV1)是美国鲶鱼养殖业中最重要的病毒病原体。人们对 IcHV1 的遗传稳定性和抗原变异性知之甚少。本文通过限制性片段长度多态性(RFLP)分析和血清中和试验评估了 IcHV1 现场分离物的遗传和抗原多样性。RFLP 分析确定了两种不同的基因型(IcHV1A 和 IcHV1B),它们都是从蓝鲶异型疱疹病毒(BCAHV)中分离出来的。用抗 IcHV1 单克隆抗体 Mab-95 进行的中和试验表明,IcHV1A 和 IcHV1B 有共同的抗原决定簇,而 BCAHV 则没有,Mab-95 也不能中和 BCAHV。对具有代表性的分离物进行的毒力评估表明,RFLP 组内的分离物之间存在显著差异,RFLP 组的集合数据表明 IcHV1B(集合存活率[平均值±SE]:58.3%±2.6)可能比 IcHV1A(存活率:68.6%±2.4)更具毒力。用具有代表性的 IcHV1A 和 IcHV1B 分离物进行再挑战表明,存在交叉保护效应,初次暴露于 IcHV1A 或 IcHV1B 的鱼类在随后再次暴露于 IcHV1A 或 IcHV1B 时会表现出很强的保护能力。这项工作证明了病例分离物之间在毒力上的显著差异,确定了两个不同于 BCAHV 的独立 IcHV1 系,它们在河豚和河豚 × 蓝鮰杂交种中具有相似的毒力,并且在暴露于任一系的鲶鱼中都具有交叉保护作用。
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引用次数: 0
Medicinal herbs, bioactives, phytochemicals and functional foods for health of aquatic species: Exploring a burgeoning focus for fish health sciences. 促进水生物种健康的药草、生物活性物质、植物化学物质和功能食品:探索鱼类健康科学的新兴重点。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-08-04 DOI: 10.1111/jfd.13999
David J Speare, Barbara F Nowak, Heike Schmidt-Posthaus, Saengchan Senapin
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引用次数: 0
Multiplex PCR assay for the accurate and rapid detection and differentiation of Lactococcus garvieae and L. petauri 用于准确、快速检测和区分 Garvieae 乳球菌和 Petauri 乳球菌的多重 PCR 法。
IF 2.2 3区 农林科学 Q2 FISHERIES Pub Date : 2024-08-04 DOI: 10.1111/jfd.14004
Dilek Ustaoglu, Rafet Çağrı Öztürk, Mustafa Ture, Silvia Colussi, Paolo Pastorino, Ana Isabel Vela, Charalampos Kotzamanidis, Donatella Volpatti, Pier Luigi Acutis, Ilhan Altinok

Lactococcosis is a common bacterial fish disease caused by Lactococcus garvieae, L. petauri and L. formosensis. Although there are different PCR-based techniques to identify the etiological agent, none of these can differentiate these two bacteria without sequencing PCR-amplified fragments. In the present study, we developed a multiplex PCR assay for simultaneous detection and differentiation of L. garvieae and L. petauri. The specificity of the primers was validated against the bacterial DNA of the targeted and non-targeted bacteria. The sizes of the PCR amplicons were obtained as 204 bp for the DUF1430 domain-containing protein gene of L. garvieae, 465 bp for the Lichenan permease IIC component gene of L. petauri, and 302 bp for the teichoic acid biosynthesis protein F gene of both L. garvieae and L. petauri. The PCR amplicons were clearly separated by agarose gel electrophoresis. The multiplex PCR assay did not produce any amplification products with the DNA of the non-targeted bacteria. The multiplex PCR detection limits for L. garvieae and L. petauri were 5 and 4 CFU in pure culture and 50 and 40 CFU/g in spiked tissue samples, respectively. It takes less than 2 h from plate-cultured bacteria and 3 h from tissue samples to get results. In conclusion, the developed multiplex PCR assay is a rapid, specific, accurate, and cost-effective method for the detection and differentiation of L. garvieae and L. petauri and is suitable to be used for routine laboratory diagnosis of L. garvieae and L. petauri.

乳球菌病是一种常见的细菌性鱼病,由 Lactococcus garvieae、L. petauri 和 L. formosensis 引起。虽然有不同的基于 PCR 的技术来确定病原体,但如果不对 PCR 扩增片段进行测序,这些技术都无法区分这两种细菌。在本研究中,我们开发了一种多重 PCR 检测方法,用于同时检测和区分 L. garvieae 和 L. petauri。针对目标细菌和非目标细菌的细菌 DNA 验证了引物的特异性。PCR扩增子的大小分别为:L. garvieae的含DUF1430结构域的蛋白基因为204 bp,L. petauri的Lichenan渗透酶IIC成分基因为465 bp,L. garvieae和L. petauri的茶色酸生物合成蛋白F基因均为302 bp。PCR 扩增子在琼脂糖凝胶电泳中被清楚地分离出来。多重 PCR 检测未与非目标细菌的 DNA 产生任何扩增产物。在纯培养物中,L. garvieae 和 L. petauri 的多重 PCR 检测限分别为 5 CFU 和 4 CFU,在加标组织样本中分别为 50 CFU 和 40 CFU/g。平板培养细菌和组织样本的检测结果分别需要不到 2 小时和 3 小时。总之,所开发的多重 PCR 法是一种快速、特异、准确且经济有效的方法,可用于检测和鉴别 L. garvieae 和 L. petauri,适合用于 L. garvieae 和 L. petauri 的常规实验室诊断。
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引用次数: 0
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Journal of fish diseases
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