传染性肌肉坏死病

2008年农业部兽医局委托动物流行病学中组织专家编写动物疫病释义,为便于解读水生动物的疫病,本刊现将《一、二、三类动物疫病病种名录》中水生动物疫病种类的分类及各病的释义分期进行刊登。     

鲍病毒性死亡病

鲍病毒性死亡病,也称为鲍病毒病、鲍裂壳病,是严重威胁我国南北鲍养殖的一种疾病.为我国水生动物三类疫病.一、病原学病原是一些鲍球形病毒(Abalone spherical viruses),其分类地位不详.包括四种球形病毒:第一种直径90nm～140nm,有二层囊膜(8nm～10nm)和光滑的表面,核衣壳直径70nm～100nm.在血细胞或者结缔组织的细胞质里复制;第二种直径100nm,有囊膜.核衣壳为六边形(或二十面体),在肝、肾和肠道的上皮细胞质里复制,通常在内质网里,是DNA病毒;第三种直径135nm～150nm纳米,有囊膜,表面有突起.二十面体的核衣壳直径为100nm～110nm.在受感染的肝、肾细胞(包括上皮细胞和结缔细胞)的细胞质中有双层膜的空泡里装配;推测它们在细胞核里复制;第四种直径90nm～110nm,有光滑表面的囊膜,二十面体的核衣壳,在受感染的肝、肾细胞(包括上皮细胞和结缔细胞)的细胞质中有双层膜的空泡里装配二十面体的核衣壳;推测它们在细胞核里复制.     

水生动物疫病病种介绍：传染性造血器官坏死

2008年农业部兽医局委托动物流行病学中组织专家编写动物疫病释义,为便于解读水生动物的疫病,本刊现将《一、二、三类动物疫病病种名录》中水生动物疫病种类的分类及各病的释义分期进行刊登。     

黏孢子虫病

2008年农业部兽医局委托动物流行病学中组织专家编写动物疫病释义,为便于解读水生动物的疫病,本刊现将《一、二、三类动物疫病病种名录》中水生动物疫病种类的分类及各病的释义分期进行刊登。     

小瓜虫病Ichthyophthiriasis

2008年农业部兽医局委托动物流行病学中组织专家编写动物疲病释义,为便于解读水生动物的疫病,本刊现将《一、二、三类动物疫病病种名录》中水生动物疫病种类的分类及各病的释义分期进行刊登。     

闽南地区养殖石斑鱼病毒性神经坏死病初步研究

应用流行病学调查方法。对发生在福建南部海区网箱养殖石斑鱼的暴发性传染病进行了初步研究。应用PCR和RT-PCR技术。检测石斑鱼常见的病毒性神经坏死病病毒、真鲷虹彩病毒，结果各发病渔场均检出病毒性神经坏死病病毒；病原菌分离未发现强致病性细菌；寄生虫检查发现腹腔普遍有一种微孢子虫感染形成的孢囊。临床症状主要表现为游动失常、厌食、鳔肿大、肾脏坏死、糜烂。表明病毒性神经坏死病病毒感染是导致石斑鱼大批死亡的主要原因。     

鲍脓疱病

2008年农业部兽医局委托动物流行病学中心组织专家编写动物疫病释义,为便于解读水生动物的疫病,本刊现将《一、二、三类动物疫病病种名录》中水生动物疫病种类的分类及各病的释义分期进行刊登。     

白斑综合征

2008年农业部兽医局委托动物流行病学中组织专家编写动物疫病释义,为便于解读水生动物的疫病,本刊现将《一、二、三类动物疫病病种名录》中水生动物疫病种类的分类及各病的释义分期进行刊登。     

草鱼出血病

2008年农业部兽医局委托动物流行病学中组织专家编写动物疫病释义,为便于解读水生动物的疫病,本刊现将《一、二、三类动物疫病病种名录》中水生动物疫病种类的分类及各病的释义分期进行刊登。     

折光马尔太虫病

折光马尔太虫病,是由折光马尔太虫寄生于牡蛎、贻贝等双壳类动物消化系统的一种寄生虫病,牡蛎染疫后表现虚弱、消瘦、生长停滞和高死亡率.为OIE疫病和我国水生动物三类疫病.     

Viral nervous necrosis (VNN) associated with mass mortalities in cage-reared sea bass,Dicentrarchus labrax (L.)

Mass mortalities of the European sea bass, Dicentrarchus labrax (L.), occurred in different ongrowing units in Greece. A presumptive diagnosis of viral nervous necrosis (VNN) was made on the basis of the light microscopic observations of a vacuolating encephalopathy and retinopathy. Positive peroxidase reaction with VNN antiserum confirmed nodavirus as the causative agent. Usually recorded in larvae and juvenile stages of different marine fish species, VNN is described in adult sea bass and sea bass reared in floating cages in the Mediterranean for the first time. The horizontal transmission of the disease has been strongly suspected. Histological analyses clearly demonstrate that cells other than nerve cells are also infected. Some epidemiological aspects of the disease are described and their implications for the establishment of prophylactic guidelines are discussed.     

Viral nervous necrosis in hatchery-reared larvae and juveniles of Japanese parrotfish, Oplegnathus fasciatus (Temminck & Schlegel)

Abstract. Mass mortalities of hatchery-reared Japanese parrotfish larvae and juveniles, Oplegnathus fasciatus (Temminck & Schlegel), have occurred in Nagasaki Prefecture. Light and electron microscopic examinations showed that the only consistent histopathological feature was extensive nervous necrosis in the spinal cord, spinal ganglia and brain. Numerous non-enveloped virus particles, icosahedral in morphology and measuring about 34 nm in diameter, were found in the cytoplasm of affected neurones and glial cells. Such nervous necrosis is believed to be the major cause of the mass mortalities of hatchery-reared Japanese parrotfish larvae and juveniles.     

鱼类神经坏死病毒研究进展与发展趋势

神经坏死病毒(nervous necrosis virus,NNV)是一种世界范围内流行、严重危害多种海水和淡水鱼类的传染性病原。NNV为单一正链、2节段RNA病毒,基因组由RNA1(3.1 kb)和RNA2(1.4 kb)组成。在病毒复制过程中,会合成亚基因组RNA3。RNA1编码RNA聚合酶。RNA2编码衣壳蛋白,为病毒的唯一结构蛋白。RNA3编码B1和B2两种非结构蛋白。根据病毒衣壳蛋白的基因序列,神经坏死病毒可以分成4种基因型,分别为拟鲹、红鳍东方鲀、条斑星鲽和赤点石斑神经坏死病毒基因型。但是,目前只发现A、B、C三种病毒血清型,A对应拟鲹神经坏死病毒基因型,B对应红鳍东方鲀神经坏死病毒基因型、C对应条斑星鲽神经坏死病毒和赤点石斑神经坏死病毒基因型。病毒存在垂直和水平两种传播途径,而且广泛分布于养殖和野生鱼类中。阻断病毒在野生与养殖鱼类之间的传播和开展新型鱼类疫苗研发是将来研究趋势。     

Characterization of grouper nervous necrosis virus (GNNV)

Grouper nervous necrosis virus (GNNV) was isolated from moribund grouper larvae, Epinephelus sp., using a fish cell line GF-1. The present study describes the biochemical and biophysical properties of GNNV and the expression of GNNV in diseased grouper larvae. Viral protein was detectable in most of the GNNV-infected GF-1 cells by the fluorescent antibody technique (FAT) after 12 h post-infection (p.i.), although no cytopathic effect (CPE) appeared at that time. Clear CPE developed on the third day, and complete disintegration of the monolayer occurred over the subsequent two days. The infectivity of GNNV can be blocked following treatment at 60 °C for 1 h. GNNV was sensitive to pH 3 and pH 10–12 with a 4 log₁₀ drop in infectivity. Purified GNNV was analysed by SDS–PAGE, and then stained with periodic acid silver. The positive staining indicated that its two capsid proteins were glycoproteins. Genomic RNAs of GNNV were extracted from purified virions and analysed. The molecular weights of genomic RNAs were 1.02 × 10⁶ and 0.50 × 10⁶ Da. The T2 region of the coat protein gene of GNNV was amplified by polymerase chain reaction (PCR), and the multiple alignment of the T2 sequence of two GNNV isolates with four genotypes of fish nodaviruses revealed that these two isolates (GNNV9410 and GNNV9508) belong to the red-spotted grouper nervous necrosis virus (RGNNV) genotype. The tissue distribution of GNNV in naturally infected grouper larvae was investigated by in situ hybridization using a dig-labelled probe, which showed that GNNV was not only detected in the brain and retina, but also in the gill, skeletal muscle, liver, pyloric gland, intestine and blood cells in the heart.     

High prevalence of betanodavirus barfin flounder nervous necrosis virus as well as red‐spotted grouper nervous necrosis virus genotype in shellfish

Using two serially executed PCRs, the discriminative multiplex two‐step RT‐PCR (DMT‐2 RT‐PCR) following the detection seminested two‐step RT‐PCR (DSN‐2 RT‐PCR), we found a high frequency presence of BFNNV genotype as well as RGNNV in various domestic and imported shellfish. This was definitely different from the previous reports of outbreaks and asymptomatic infection only by the RGNNV genotype in cultured finfish in Korea. Cultivation of NNV entrapped in shellfish was performed successfully by a blind passage. Thus, in an attempt to elucidate the epidemiology of betanodavirus, experiments conducted on 969 shellfish samples concluded that (i) distribution of NNV genotype, especially BFNNV, in shellfish is clearly different from that found in finfish of the world; (ii) unlike RGNNV, which showed a high rate in summer, BFNNV showed no seasonal variation and this result suggests BFNNVs in the marine environment remain fairly constant throughout the year; and (iii) the entrapped virus in shellfish was alive and culturable in vitro. These results are the first report of high level prevalence of in vitro culturable NNV in shellfish, for both BFNNV and RGNNV, which may present a potential risk in transmitting nodaviruses to host species in a marine environment.     

神经坏死症病毒对长缟鲹仔鱼的感染及在体内的传播

以长缟鲹(Pseudocaranx dentex)孵化仔鱼为材料，通过神经坏死病毒(Nervous Necrosis Virus，NNV)浸浴感染实验，明确病毒浓度越高，供试鱼被病毒感染的时间越早，病毒检出率越高；运用细胞组织病理学和酶联免疫(Enzyme-1inked immunosorbent assays，ELISA)的原理和方法，了解了神经坏死病毒的侵入、感染和在神经组织中的扩散方式；利用ELISA技术，结合电子显微镜对NNV感染细胞进行超显微观察，确认育苗水环境中神经坏死病毒可水平感染供试鱼皮肤上皮的基底细胞；病毒感染细胞发生细胞质内质网膨胀、细胞核变性、细胞小器官数减少等病理变化；病毒感染细胞质中病毒粒子有散布、类结晶和二者兼有3种存在形式。本研究成果可为了解海水鱼类人工育苗的病毒性神经坏死症感染途径、传播和预防病毒性神经坏死症的发生提供科学依据。     

鱼类神经坏死病毒实时荧光RT-PCR检测方法的建立和应用

根据GenBank中登录的鱼类神经坏死病毒CP基因序列,选择高度保守区域设计引物和TaqMan荧光探针,通过对实时荧光RT-PCR反应条件进行优化,建立了用于检测鱼类神经坏死病毒的实时荧光RT-PCR方法。利用该方法检测鱼类神经坏死病毒及其他多种常见的水生动物RNA病毒,结果只能检测到目的病毒,表明其具有良好的特异性。灵敏性试验发现,其最低检测限可达1．2pg／μL的总RNA。与RT-PCR的灵敏度对比试验表明,其敏感度比RT-PCR高100倍。对同一样品进行检测,在组内及组间的变异系数分别为0．9％以及1．5％,证实其重复性极好,并且从抽提核酸到得出结果仅需4h。对临床500份样品进行鱼类神经坏死病毒检测,结果发现有40份阳性样品。这些结果表明,本研究所建立的实时荧光RT-PCR能对鱼类神经坏死病毒进行准确、快速的检测,具有特异性好、灵敏度高的优点,是开展鱼类神经坏死病的临床检测和疫情监测工作的有力工具。[中国水产科学,2008,15（3）：506-510]     

Age dependency of nervous necrosis virus infection in barramundi Lates calcarifer (Bloch)

Age‐dependent susceptibility to nervous necrosis virus (NNV) was demonstrated for barramundi (Lates calcarifer). The experiment used juvenile barramundi produced from a single spawning that were challenged consecutively by immersion with a redspotted grouper nervous necrosis virus (RGNNV) isolate. The dose and environmental conditions (35 ppt salinity and 30 °C) were constant. Fish and water were sampled longitudinally for histopathology and RT‐qPCR analysis to examine the evolution of the disease, virus replication, immune response and release of virus into water. Viral nervous necrosis (VNN) disease occurred in barramundi challenged at 3 and 4 weeks of age while fish challenged at 5, 7 and 9 weeks of age developed subclinical infection. Replication of NNV occurred faster and the concentration of virus reached higher concentrations in the younger fish with clinical disease. Virus isolation and qPCR tests indicated that infectious NNV was released from carcasses into water when fish were affected with clinical disease but not when NNV infection was subclinical. Based on these observations, we consider that carcasses from clinically infected fish have a potentially important role in the horizontal transmission of NNV, and barramundi juveniles should be protected from exposure to NNV until they are 5 weeks of age and reach the disease resistance threshold.     

Protective immunity of sevenband grouper, Epinephelus septemfasciatus Thunberg, against experimental viral nervous necrosis

This paper describes the protective immune responses of sevenband grouper, Epinephelus septemfasciatus Thunberg, immunized with live piscine nodavirus, the causative agent of viral nervous necrosis (VNN), or the Escherichia coli – expressed recombinant coat protein. Nodavirus-neutralizing antibodies were detected at titres ranging from 1:158 to 1:1257 in serum of sevenband grouper which survived intramuscular injection with the virus, by a cell culture assay system. The virus-neutralizing ability of immune serum was also confirmed by injecting virus previously treated with serum into fish. This indicates establishment of acquired immunity in survivors and thus explains why survivors from natural infection are resistant to recurrence of the disease. Young sevenband grouper were immunized twice by intramuscular injections with the recombinant coat protein. Immunized fish produced neutralizing antibodies at high titres for at least 110 days and showed significantly lower mortalities in virus challenge tests. These results suggest the potential for vaccination against VNN in sevenband grouper, which is susceptible to piscine nodavirus at all life-stages.