小杂鱼携带细菌与大菱鲆疾病发生的相关性

腹水病是工厂化养殖大菱鲆的一种常见病和多发病，常造成鱼类大规模的死亡。2006年6月山东某大菱鲆Scophthalmus maximus养殖场发生大规模腹水病，通过对病鱼细菌的分离、纯化和培养，对养殖井水、养殖池水和所用饵料小杂鱼进行细菌追踪调查，将从该养殖场所用小杂鱼中分离得到的细菌RF-1与鱼体分离的细菌FS-1进行比较，并对其进行人工感染试验，结果证明这2株菌对健康大菱鲆都具有很强的致病性，而且感染症状完全一致。进而对这2株菌进行了API—ID32E鉴定、传统的生理生化鉴定和16S rRNA序列分析，发现小杂鱼携带细菌RF-1与病原菌FS-1具有一致性，均为溶藻弧菌（Virio alginolyticus）。最后对这2株细菌进行了药敏试验，为控制疾病的发生和蔓延提供用药指导。     

中国大菱鲆虹彩病毒(TRBIV)PCR检测方法的建立及其应用

大菱鲆红体病虹彩病毒(turbot reddish body iridovirus,TRBIV)是中国工厂化养殖大菱鲆的主要病毒性病原之一.本研究提取了该病毒DNA,依据其主要衣壳蛋白基因序列设计了PCR引物,优化了PCR反应参数,建立了TRBIV的PCR检测方法,测试了该方法的特异性和灵敏度,并应用该方法开展了TRBIV的流行情况调查及研究了大菱鲆的外观症状(红体)与TRBIV感染的关系.结果显示,本研究建立的TRBIV PCR检测方法可以从相当于100ng病鱼组织中或103数量级的病毒粒子中检出TRBIV,也可以在不杀死被测鱼的情况下,仅从鱼体中抽取少量血液即可在1天的时间内完成大量样品的TRBIV检测,但不能从健康大菱鲆脾组织和患淋巴囊肿牙鲆的囊肿组织中扩增出任何产物,说明该方法具有很高的灵敏性和特异性;在所调查的山东半岛5家大菱鲆养殖场的19尾大菱鲆中,7尾为TRBIV阳性或弱阳性;具有"红体"症状的大菱鲆应当划分为"病毒性红体病"、"细菌性红体病"和"非病原性红体症"3种不同的类型.本研究为TRBIV的流行情况和传播途径调查、疾病的快速诊断和控制提供了技术手段;调查结果显示TRBIV已遍布山东半岛沿海地区的大菱鲆主产区,在地域上相距较远的多个大菱鲆养殖场流行,今后需要密切关注该病毒的传播和流行.     

大菱鲆常见细菌病的诊治技术

近年来大菱鲆养殖在我国发展迅猛，在山东等环渤海地区大规模养殖。由于目前国内大菱鲆养殖场家苗种质量参差不齐，开放式流水养殖而影响大环境质量，大部分场家直接投喂自己加工的冰鲜杂鱼饲料，导致疾病的传播机会增多，加之管理不科学使病害感染机会增多。经过近几年的观察，细菌病成为大菱鲆养殖危害最严重的     

大菱鲆的常见病害及对策（下）

7．白便症 病原为大菱鲆弧菌和溶藻胶弧菌。病鱼体色变暗，腹部下凹，不摄食或吞食后吐出，挤压腹部可见白便从肛门流出，有时肛门处拖带稠的白色粪便。因此，发病时池底经常会发现黄白色条状物。该病常见于育苗期和养成期，其感染率及死亡率均较高，能引起长期性慢性死亡，养殖过程中常与腹水病状并发。     

养殖大菱鲆肠道中大菱鲆弧菌的分离鉴定及药敏试验

2014年11月,大连地区某大菱鲆养殖场发生病害并伴有死亡,主要症状为吻部下颌明显出血,腹腔积水,肝脏充血,肠道出血有白便。从患病大菱鲆肠道中分离出1株可在硫代硫酸盐—柠檬酸盐—胆盐—蔗糖琼脂培养基上生长的菌株HZ-C1,人工回接感染试验证明其对健康大菱鲆具有较强致病性。通过形态学观察、16SrDNA序列同源性分析及生理生化试验确定菌株HZ-C1为大菱鲆弧菌。药敏检测结果显示,大菱鲆弧菌HZ-C1对氯霉素、丁胺卡那及头孢曲松等11种抗生素敏感,但对青霉素、四环素及诺氟沙星等15种抗生素耐药。本研究为大连地区养殖大菱鲆细菌性病害防治提供了一定病原学依据。     

弧菌对养殖大菱鲆的危害及其防治方法研究新进展

弧菌是细菌性疾病的常见病原,不但会感染大菱鲆(Scophthalmus maximus),还会对人类健康造成重大危害。大菱鲆是我国重要的海水养殖鱼类,具有较高的经济价值,然而疾病问题严重阻碍了其养殖业的发展。近年来,有关大菱鲆弧菌病检测及防治方面的研究有了新的突破。文章介绍了几种常见弧菌病原对养殖大菱鲆的危害,对新的检测及防治方法进行了总结,可为大菱鲆弧菌病的进一步研究提供借鉴。     

7种消毒剂对大菱鲆烂鳍病致病原鳗弧菌的杀灭作用研究

大菱鲆（Scophtbalmus maximus）养殖遍及辽宁、河北、山东、福建等地，成为我国北方沿海海水养殖的支柱产业之一，年产量4万～5万t，产值30多亿元，产生了良好的经济和社会效益。随着养殖规模的迅速扩大，大菱鲆疾病问题日渐突出。其中，烂鳍病是大菱鲆养殖中一种较为严重的细菌性疾病，流行范围极广，几乎在各个养殖场均有发生，危害较大。大菱鲆从苗期、养成期直到亲鱼培育期均可感染此病，累积死亡率高达80％-90％。     

我国养殖大菱鲆病毒性红体病及其流行情况调查

大菱鲆病毒性红体病(Viral reddish body syndrome,VRBS)是一种新发现的感染我国养殖大菱鲆的流行性疾病。本文描述了病鱼的外观症状和解剖病理特征,报道了该病的病原及疾病流行情况调查结果。外观检查发现,病鱼的体表无明显损伤,但腹面沿脊椎骨附近皮下淤血、发红,鳍及鳍基部充血、发红;病鱼贫血,血液凝固性差;肾脏肿大,呈灰白色。组织病理学研究显示病鱼脾组织中存在大量肥大细胞,电镜切片中可见大量平均直径约125nm的二十面体病毒粒子,即大菱鲆红体病虹彩病毒(Turbot reddish body iridovirus,TRBIV)。将过滤除菌的含TRBIV的病鱼脾组织匀浆液,经腹腔注射进行人工感染,感染鱼在3周内的累积死亡率达85．7％,死亡大菱鲆表现出腹面和鳍边发红等外观症状,并在感染鱼脾组织切片中观察到大量同样的病毒粒子,由此证实TRBIV是我国养殖大菱鲆病毒性红体病的病原。疾病流行情况调查显示,该病多在养成期大菱鲆中流行,高发季节为每年的8～12月。     

天津地区观赏鱼养殖新发疾病调查研究

<正>2012年至2013年期间,天津市水产技术推广站的技术人员就天津地区观赏鱼养殖疾病发生情况进行了取样调查。两年间共采集病样210例,通过实验室检测发现,观赏鱼养殖疾病以细菌病和寄生虫病为主,占调查病例的80%以上,且多为细菌和寄生虫的混合感染,真菌病所占比例较少,未发现病毒病,其它非生物性病原疾病占16.7%。确定27种观赏鱼疾病,其中真菌引起的疾病1种,为水霉病;寄生虫引起的疾     

大菱鲆脾肾结节病病原菌的分离和鉴定

2017年夏季，江苏省连云港市某养殖场大菱鲆出现大规模死亡，发病鱼体表无明显症状，解剖可见脾脏、肾脏出现白色散在结节，从发病鱼的内脏中分离得到1株优势菌SM-Myco001。人工感染实验结果显示，SM-Myco001可以引起大菱鲆内脏结节症状并造成鱼类死亡，且在高养殖水温(22℃)条件下发病更为剧烈。生理生化鉴定和16S rRNA基因分析结果显示，SM-Myco001属于海分枝杆菌(Mycobacterium marinum)。本研究首次报道了我国养殖大菱鲆感染海分枝杆菌的病例，可为大菱鲆养殖过程中的疾病防控提供参考。     

Ocular bacterial signatures of exophthalmic disease in farmed turbot (Scophthalmus maximus)

As is the case at other sites in the body (e.g. the gut, skin and mouth), the ocular microbiota plays a crucial role in their host, as disturbances of the composition and function of the ocular microbiota are known to be associated with ocular disorders. Exophthalmic disease (ED) is a significant cause of high mortality in fish species, including farmed turbot (Scophthalmus maximus). However, the relationship between alterations in the ocular microbiota and ED in turbot is unclear. In this work, we collected turbot samples from farmed ponds with ED and healthy samples to understand changes in the ocular microbiota of turbot suffering from ED. We compared the structural and metabolic differences of ocular bacterial communities from farmed turbot with exophthalmic disease and those of healthy controls. Besides less microbial diversity found in turbot with ED regarding the control group, we also found that Aeromonas was the dominant bacteria both in controls and ED samples, but the abundance of Aeromonas was significantly greater in ED individuals. Moreover, the results of correlation test further suggest that Aeromonas overgrowth was correlated with the progress of the disease and shifts in ocular microbiota functional pathways in turbot. These findings emphasize that an increased abundance of Aeromonas serves as an ocular bacterial signature associated with ED in turbot, which provide basic information useful for diagnoses, prevention and treatment of ocular diseases occurring in cultured fish.     

大菱鲆仔稚鱼发育早期肠道菌群结构形成的影响因素

为研究鱼类发育早期肠道菌群结构的演变过程及影响因素,运用高通量测序技术,分析了处于不同发育阶段的大菱鲆(Scophthalmus maximus)仔稚鱼肠道、受精卵、不同类型的饵料和水源中的菌群结构,以及它们之间的相关性。结果显示,以不同的OTU(Operational Taxonomic Units,可操作分类单元)作为分类依据,发现大菱鲆仔稚鱼的肠道菌群结构在开口摄食后不久已趋于稳定,其优势菌与受精卵所携带的细菌关联较大。并且在大菱鲆仔稚鱼不同的发育时期,这一菌群的结构非常稳固,几乎不受水和饵料中优势细菌的影响而发生改变。乳球菌属的Lactococcus piscium菌株一直是大菱鲆仔稚鱼肠道中的优势菌种,在不同发育时期的优势度高达45%~65%。本研究还发现,大菱鲆仔稚鱼肠道可能对定植的菌种具有选择性,一些水环境和饵料中的非优势菌,如Streptococcus sp.,Pseudomonas sp.,Carnobacterium sp.等细菌也会定植于肠道,成为大菱鲆肠道中的次优势菌。     

A study of the susceptibility of Atlantic halibut,Hippoglossus hippoglossus (L.), to viral haemorrhagic septicaemia virus isolated from turbot,Scophthalmus maximus (L.)

The susceptibility of Atlantic halibut, Hippoglossus hippoglossus (L.), to viral haemorrhagic septicaemia virus (VHSV) was tested. Juvenile halibut of approximately 5 g weight were subjected to challenge by intraperitoneal injection, cohabitation and immersion to a VHSV isolate from an outbreak of the disease in turbot, Scophthalmus maximus (L.). The intraperitoneal injection gave the highest mortality rate of 28% after 50 days. The cohabitee group suffered 19% mortality rate and the immersion group only 2%. Control groups included turbot exposed either by intraperitoneal injection or immersion which suffered mortality rates of 93 and 50%, respectively. The results suggest that halibut are markedly less susceptible to VHSV than turbot.     

养殖大菱鲆主要疾病及防治技术

在流行病学调查的基础上，对目前我国大菱鲆养殖中常见疾病进行了较全面的介绍，包括病毒性、细菌性和寄生虫性疾病。系统地描述了这些疾病的流行特征、发病症状、危害程度、感染率及致病原等。同时为各种疾病提供了具体的防治技术和措施，对大菱鲆养殖生产与疾病防治具有理论指导意义和重要的参考价值。本文系国内首次对养殖大菱鲆疾病进行的全面性报道。     

Study of the distribution of active caspase‐3‐positive cells in turbot,Scophthalmus maximus (L.), enteromyxosis

Enteromyxosis caused by Enteromyxum scophthalmi is one of the parasitizations with a higher economic impact on turbot, Scophthalmus maximus (L.), aquaculture. This myxosporean produces severe catarrhal enteritis with abundant inflammatory infiltrates in the lamina propria‐submucosa (LP), epithelial detachment and leucocyte depletion of the lymphohaematopoietic organs. Some advances made on the pathogenesis pointed to a role of apoptosis in the enteromyxosis. Therefore, the main aim of this work was to employ the TUNEL assay and the anti‐(active caspase‐3) immunohistochemical assay to detect apoptotic cells in both healthy and E. scophthalmi‐infected turbot in order to establish the presence and distribution of apoptotic cells during development of the disease. More apoptotic cells located within the gastrointestinal epithelium were observed in the initial stages of the infection in E. scophthalmi‐infected turbot compared with non‐infected turbot. As the infection progressed, a higher degree of apoptosis occurred in the epithelium of folds heavily parasitized. In the severely infected turbot, apoptosis was also found among the leucocytes of the intestinal inflammatory infiltrates. Moreover, the number of active caspase‐3‐positive cells in the lymphohaematopoietic organs tended to increase with disease severity. In view of the results, increased apoptosis in the epithelium may favour the scaling that occurs during enteromyxosis and cell death of leucocytes in the intestinal LP, contributing to leucocyte depletion in severe cases.     

Streptococcosis in cultured turbot, Scopthalmus maximus (L.), associated with Streptococcus parauberis

Several strains of Garm-positive short rod (coccibacilli)-shaped bacteria were isolated from diseased cultured turbot, Scophthalmus maximus (L.), in the North of Spain with lesions and signs of Spain with lesions and signs of streptocecosis. The α-haemolytic streptoceoccus-like organisms from diseased turbot were identified by physiological, biochemical and 16S rRNA gene sequence analysis as Streptococcus parauberis. This is the first report of S. parauberis associated with fish disease.     

有水与无水保活运输对大菱鲆生理应激及鱼肉品质的影响

为探究大菱鲆分别在有水及无水条件下运输过程中生理生化指标及肌肉品质的变化,为大菱鲆的保活运输提供参考。将大菱鲆进行暂养、降温、充氧包装后进行模拟运输,并在运输不同时间段取样,检测大菱鲆生理生化指标及肌肉指标。结果显示,模拟运输18 h后,有水运输组和无水运输组中的大菱鲆存活率仍为100%。大菱鲆肌肉中水分含量、粗脂肪、pH、糖原、三磷酸腺苷(ATP)和腺苷酸(AMP)含量显著降低,乳酸和肌苷酸(IMP)含量升高。其中,有水运输组中的大菱鲆AMP含量降低了38.65%,IMP含量增加了29.1%,无水运输组中的大菱鲆AMP含量降低了12.83%,IMP含量增加了28.47%。大菱鲆血液生化指标中,葡萄糖、尿素氮和皮质醇含量显著升高。运输过程后,无水运输组中大菱鲆生理应激及能量代谢指标的变化幅度均小于有水运输处理组中的大菱鲆。此外,低温无水运输组大菱鲆肌肉中呈味核苷酸IMP和AMP含量显著高于有水运输组,表明低温无水运输增加了大菱鲆呈鲜味物质的含量,提高了鱼肉风味。研究表明,低温无水保活技术在活鱼运输中具有广泛的应用前景。     

Pathology of Enterococcus sp. infection in farmed turbot, Scophthalmus maximus L.

Abstract The lesions of an enterococcal septicaemia occurring in turbot, Scophthalmus maximus (L.), cultured in several farms in northwest Spain are described. The agent of this septicaemia was identified previously as an Enterococcus -like bacterium. Two main patterns of lesions were observed in the diseased turbot: a focal form characterized by exophthalmia, muscular haemorrhages, acute branchitis, and suppurative inflammation of periorbital tissues, eyeball, meninges and brain; and another generalized form with similar lesions, but showing more extensive haemorrhages, ulceration and purulent inflammation of the skin, desquamative enteritis, and necrosis of spleen and kidney. Fatty changes of the liver were observed in all turbot examined (diseased and controls), which indicates that these alterations were not related to the infection. Experimentally inoculated turbot developed lesions corresponding to the generalized form of the natural disease. Although the Enterococcus strain was recovered in pure culture from all the organs, Gram-positive bacteria could not be visualized histologically in the lesions of muscle and brain.