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1.
鱼类诺达病毒及其所导致的疾病   总被引:2,自引:1,他引:1  
黄剑南 《水产学报》2006,30(6):831-836
In recent years, piscine nodaviruses have emerged as major pathogens of a wide range of larval and juvenile marine finfish resulting in high mortality in aquaculture worldwide. Affected fish exhibit a range of neurological signs, such as erratic swimming behaviour with the associated microscopic lesions of necrosis and vacuolation of the central nervous tissues and retina. Numerous roundshaped, unenveloped and 25-30 nm in diameter virus particles were found in the cytoplasm of affected retinal and nerve cells. Nodaviruses have a bipartite genome of positivesense RNA,with RNA1 encoding the RNAdependent RNA polymerase and RNA2 encoding the capsid protein. Both RNA are capped, but not polyadenylated. The family Nodaviridae comprises two genera: Alphanodavirus and Betanodavirus, members of which primarily infect insects and fish, respectively. Therefore, betanodavirus is also named piscine nodavirus. At present, piscine nodaviruses are divided into four genotypes based on partial sequences of the coat protein gene. ELISA and RT-PCR amplification have been developed as specific diagnostic methods for the d etection of the virus. Antibodies to striped jack (Pseudocaranx dentex) nervous necrosis (SJNNV) were found in 65% of plasma samples collected from wild and domestic brood stocks of striped jack, suggesting that the virus is very prevalent. Viral antigens were detected in eggs, larvae, and ovaries of hatcheryreared and wild spawner fish, suggesting both horizontal and vertical modes of transmission of the virus. Selection of nodavirusfree spawners using ELISA for detection of antigens and RT-PCR techniques have successfully reduced incidences of the virus infections in juvenile sea bass (Dicentrarchus labrax),striped jack and barfin flounder (Verasper moseri). The SSN1 and GF cell lines have been successfully used in isolating piscinenodaviruses.Although there are many papers describing the molecular characteristics of betanodavirus, our knowledge of the genomic attributes of these viruses is still limited. Vaccination studies are being undertaken by a number of researchers and need to be fostered. In particular, the use of passive immunization of broodfish with homologous and heterologous, high titre antisera are worthy of investigation.  相似文献   

2.
美洲黑石斑鱼营养成分分析与营养价值评价   总被引:4,自引:0,他引:4       下载免费PDF全文
In this experiment, the contents of the main nutrients such as protein, fat, moisture, ash, calcium and phosphorus were analysed, and the contents of the trace elements, amino acids and fatty acids in its muscle were determined for Centropristis striata. Moreover, its nutritional value was evaluated and compared with other fishes,livestocks and poultries. The results showed that the content of dry matter and protein are higher in its muscle. Then, all of 17 common amino acids(AA) contents in muscle was 17.51% in the fresh sample, and 7 of them are human essential amino acids(EAA)41.58% in the total amino acids(TAA). The ratio of essential amino acids to nonessential amino acids(EAA/NEAA) was 84.46% ,and the essential amino acids index(EAAI) was 57.71. According to AAS and CS,which basically accorded with the issued standard value by FAO and WHO. The content of 5 delicious amino acids was 44.37% in the TAA, and the ratio of branched chain amino acids to aromatic amino acids was 2.83. The content of polyunsaturated fatty acids was 22.3% rich in its muscle also, in particular EPA and DHA. In conclusion, Centropristis striata is a species of delicious and high nutritional value,which well deserves exploitation and utilization. At the same time, this experiment was preparing for the next study to provide reference data for artificial feed and the expansion of aquaculture.  相似文献   

3.
鳗源嗜水气单胞菌主要外膜蛋白基因克隆及其表达   总被引:5,自引:0,他引:5  
欧阳岁东 《水产学报》2006,30(4):566-570
A pair of primers were designed according to the published nucleotide sequence of a putative outer membrane protein gene (omp) of Aeromonas hydrophila . With the specific primers, a target fragment about 1.1 kb was amplified from Aeromonas hydrophila ML316 via PCR .The target fragment was inserted into the linearized pGEM-T easy vector. After enzyme restriction and sequencing analysis,the nucleotide data had been further analyzed by DNAman and ClutalW software. The analysis results showed that the cloned DNA fragment had a longest open reading frame (ORF) of 1035 nt,it predicted to be encoded a 344 aa protein with the molecular weight of 36 kD. Hydrophobicity analysis suggested that the protein was highly hydrophilic, especialy at the first 24 aminoacid, this region could function as signal peptide. The homologious comparison proved the cloned gene had 96% homology to the sequence of the omp gene, and the alignment of the amino acid sequence was 98% . The recombinant plasmid was constructed with the target gene and the expressing vector pGEX-4T-1 and then was transformed into E. coli BL21(DE3)by BamH and Sal I .The fusion protein was expressed under the IPTG inducing condition,and exhibited about 62 kD in size,very close to the predicted molecular weight of GSTMOMP, furthermore,the fusion protein was specifically recognized by antiserum which raised against the major outer membrane protein of AHML316. Considering all these together, it proved that the cloned gene represented the major outer membrane protein gene of AHML316, and the expressed gene products shared identical antigenicity with the natural main outer membrane protein,and also provided technical support for developing an advanced gene engineering vaccine against Aeromonas hydrophila.  相似文献   

4.
Physico-chemical properties of FGF8 in Ctenopharyngodon idellus were analyzed by bioinformatics. The full - length grass carp FGF8 pro-polypeptide of 210 amino acids including 40 basic amino acids and 16 acidic amino acids. The signal peptide sequence is MRLIPSRLSYLFLHLFAFCYYA, and its cleavage site is located between Ala22 and Gln23. Grass carp FGF8 protein is a secreted protein since the typical character of secreted protein is that its N-domain contains a signal peptide. There are two N-glycosylation in the sites of Asn37 and Asn136, respectively, which were Asn-Phe-Thr and Asn-Tyr-Thr. The molecular weight (Mw) of grass carp FGF8 protein is 24.68 kDa. Its isoelectnic point (pl) is 10.93. The protein contains four main secondary structures: α-helix, β- sheet, β-turn and random coil.  相似文献   

5.
甲胺磷、辛硫磷对坛紫菜叶状体的生理效应   总被引:8,自引:0,他引:8  
李钦 《水产学报》2004,28(2):221-224
In this paper, the effects of methmidophos and phoxim with different concentrations and schedule on soluble protein content and chlorophyl a (Chla) content in Porphyra haintanensis were studied under experimental and ecological conditions. The results showed as follows: (1) there were some time-effect and dose-effect relationships between the pollutants and Chla contents. The Chla content of most of groups decreased with the increasing of dose and prolonging of the exposure time. In the same concentration, the toxicities of phoxim to Chla content was stronger than that of methmidophos. (2) All concentration groups in this study didn‘t show the significant time-effect and dose-effect relationships between pollutants and the soluble protein. And there was no significant difference of the soluble protein content among various groups.  相似文献   

6.
动植物蛋白源替代鱼粉研究进展   总被引:42,自引:5,他引:37  
周歧存 《水产学报》2005,29(3):404-410
With the fast development of aquaculture, fish meal needs increased in recent years, however the quantity of fish catching decreases gradually. Fishmeal is a limited feed resource, and serious concem exists on the future availability of this feedstuff for incorporation in fish diets. Undoubtedly, fish meal is well recognized as the best dietary protein source for most marine carnivorous fishes which required high dietary protein levels compared to omnivorous or herbivorous fish. Fishmeal is known for their high content of essential amino acids and fatty acids, low carbohydrates, high digestibility, low levels of anti-nutritional factors (for fresh fish meal) and is a very good source of minerals and is highly palatable. Thus fish meal is in high demand as the protein source for many formulated diets. However, production of fish meal consumes approximately 35 % of the total global fish catch, and the increasing price and potentially unstable supply in the market could be limiting factors for marine fish culture. There have been strong efforts to define and develop cost-effective protein sources that can, at least partly, substitute for expensive high-quality fish meals in least-cost feed formulations. The search for fish meal substitutes and altemative dietary protein sources is an international research priority that could be of considerable economic advantages. Therefore it is urgent task to find animal and plant protein sources in place of fish meal. Among these, plant feedstuffs have received most attention in recent years, but due to their amino acid unbalances, .presence of anti-nutritional factors and low palatability, a high level of replacement of fish meal with plant feedstuffs in omnivorous fish is generally not well accepted. This paper reviews the research status for other protein sources replacing fish meal based on available information in the literature. Animal and plant protein sources nutrient values are evaluated from the aspect of digestibility, antinutrients, physiological status and suitable supplementation.  相似文献   

7.
真鲷天然抗性相关巨噬蛋白全长cDNA的克隆与序列分析   总被引:1,自引:0,他引:1  
徐美瑜 《水产学报》2005,29(1):128-132
Natural resistance associated macrophage protein (Nmmp) is an innate resistance protein to intracellular parasites, which is expressed plentifully in macrophage ceils. Nramp has been studied in mouse, human, cattle, rainbow trout and channel catfish.However, tittle was known about the structure of Pagrus major Nramp. In order to get the complete sequence of Pagrus major Nramp, a pair of primer is designed according to a 200bp known sequence of Pagrus major Nramp cDNA. By the use of SMART RACE, the full Nramp of Pagrus major cDNA about 5 000 bp was obtained, including about 200 bp 5‘ terminal region (UTR),complete encoding region and 3‘ terminal region. There were 3 ployA signals, which showed many possibilities of cutting at 3‘ terminal region. The character of Pagrus major Nramp nucleotide sequence and deduced amino acid sequence are analyzed. 12 putative transmembrane(TM) regions, a consensus transport motif (CTM), a predicted protein kinase C phosphroylation site and three predicted N-link glycosylation sites are indicated in its deduced amino acid sequence. The ‘consense transport motif‘ CTM is located etween TM8 and TM9. Furthermore, a protein kinase C phosphroylation site and three N-link glycosylation sites were predicted. The lignment of amino acid sequences between Pagrus major Nramp cDNA and several animals is analyzed and the deduced amino acid equence of Pagrus major Nrarnp had 77.8%, 83.0%, 82.3%, 80.0%, 81.1%, 60.4%, 70.3%, 58.5%, 69.5% identity ith rainbow trout α(AAD20721), rainbow trout β(AAD20722), channel catfish(AF400108), fathead minnow (AAF01778),common carp (CABal96), mouse 1 ( AAA39838 ), mouse 2 ( AAC42051 ), human 1 ( D50403 ), human 2 ( NP - 0(106(18 ),respectively. The alignment reveals high conservation in TM and CTM regions. Analysis result makes us get familiar with the structure nd character of fish Nramp, furthermore, offers some infonnat/on for the enhancement of immunity of fish and genetic amelioration on fish breeding.  相似文献   

8.
The mechanism of modulation of the Na/H antiport was studied in erythrocytes of the tilapia, Oreochromis mossambicus. Activation of the antiport was determined through measurements of variations of intracellular pH, using a pH-sensitive, intracellular fluorescent probe 2′, 7′-bis (carboxyethyl)-5 (6)-carboxyfluorescein (BCECF). Phorbol ester (5 μM), a stimulator of protein kinase C, activated the antiport resulting in an increase of intracellular pH; a maximum of 0.125 ± 0.027 pH units over basal level (mean ± SD; n = 4–6) was reached in 20 min. The effect was blocked by protein kinase C inhibitors staurosporine and H-7, and also by the antiport inhibitor 5-(ethyl-N-isopropyl) amiloride; this demonstrates that the stimulation of the antiport in tilapia erythrocytes can take place through a protein kinase C-mediated mechanism. Vasopressin, an important osmoregulator hormone in teleosts, was able to activate the Na/H antiport, in a manner similar to phorbol ester, but its effect was only partially sensitive to staurosporine inhibition. Considering that the physiological role of the Na/H exchanger in fish is known to be different from its role in mammalian cells, these results indicate that other signal transduction mechanisms can be involved in the modulation of intracellular pH by vasopressin. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

9.
大菱鲆胚胎的玻璃化冷冻保存   总被引:3,自引:0,他引:3  
田永胜 《水产学报》2005,29(2):275-280
Vitrification solutions were examined for their suitability of cryopreservation of turbot (Seophthulnms maximus) embryos. PMPI gave higher survival rate (75.68%) than other vitrification solutions and was considered to be suitable liar the cryopreservation of turbot embryos. The freezing point of the vitrification solutions consisted of PG and MeOH in the proportion of 3:2 was measured when the vitrification solutions were cryopreserved. It showed that the vitrification solutions with a cryoprotectant concentration of over 41% have no freezing point and the freezing points of vitrification solutions containing 33.33%-40% cryprotectants were between -32.4℃ and -65.4℃, The freezing point decreased with the increase of cryoprotectant concentration. The resistance of turbot embryos at different stages to the cryoprotectants was studied, It was found that the turbot embryos from 4-5 pairs somite to tail bud were more resistant to cryoprotectants and suitable to vitrify. A live embryo was obtained after cryopreservation in liquid nitrogen for 14h and was hatched out。  相似文献   

10.
蓝太阳鱼生长激素全长cDNA的克隆与序列分析   总被引:8,自引:1,他引:7  
曹运长 《水产学报》2004,28(5):589-593
The full length cDNA encoding growth hormone of a freshwater fish, Lepomis cyanellus, (LcGH) was cloned from pituitary RNA with RT-PCR, 3‘ and 5‘ RACE (rapid amplification of cDNA ends). The LcGH cDNA (Genbank No. AY530822), about 989nt (nucleotide) long, consisted of a open reading frame with 615nt long, 5‘and 3‘untranslated regions with 93nt and 224nt long respectively, and a 57nt poly (A) tail. The DNA sequence analysis showed that there are typical Kozak sequence and polyadenylation signal. The pregrowth hormone peptide of 204aa deduced from LcGH cDNA included a putative signal peptide (17aa) locating in its Nterminal. There exist a Asn-Cys-Thr glycosylation site at amino acid 201, and 4 cysteine residues (No. 69, 177, 194, 202) that are essential to construct two S-S bonds in this pregrowth hormone peptide. Homological comparision among LcGH and other species growth hormones showed that There is high homology (more than 85%) between growth hormone of Lepomis cyanellus and that of most perciformes fish, but low homology (less than 70%) in comparison with other species such as Siluriformes and Cypriniformes fish.  相似文献   

11.
Betanodaviruses are the causative agents of viral nervous necrosis (VNN) or viral encephalopathy and retinopathy (VER) in cultured marine fish. Based on the RNA2 gene fish nodaviruses have been traditionally classified into four different genotypes and recently a fifth genotype has been proposed. This study presents sequencing data of 24 new nodaviruses obtained from three different fish species: sea bass, Dicentrarchux labrax (L.), sea bream, Sparus aurata L., and Senegalese sole, Solea senegalensis Kaup, cultured in the Iberian Peninsula (Spain and Portugal). Sequence analysis was performed on the T4 region (388 nt) of the coat protein gene. In addition, phylogenetic analysis, according to maximum parsimony and neighbour-joining methods, was performed using these sequences and other nucleotide sequences available in the databases or in the literature. Results obtained indicate that all these new nodaviruses should be classified into the striped jack nervous necrosis virus (SJNNV) genotype. This finding suggests that SJNNV genotype is emerging in the Iberian Peninsula and could easily spread throughout the Mediterranean, representing a serious threat to the fish farming industry.  相似文献   

12.
Viral encephalopathy and retinopathy (VER), otherwise known as viral nervous necrosis (VNN), is a severe pathological condition caused by RNA viruses belonging to the Nodaviridae family, genus Betanodavirus. The disease, described in more than 50 fish species worldwide, is considered as the most serious viral threat affecting marine farmed species in the Mediterranean region, thus representing one of the bottlenecks for further development of the aquaculture industry. To date, four different genotypes have been identified, namely red‐spotted grouper nervous necrosis virus (RGNNV), striped jack nervous necrosis virus (SJNNV), tiger puffer nervous necrosis virus and barfin flounder nervous necrosis virus, with the RGNNV genotype appearing as the most widespread in the Mediterranean region, although SJNNV‐type strains and reassortant viruses have also been reported. The existence of these genetically different strains could be the reason for the differences in mortality observed in the field. However, very little experimental data are available on the pathogenicity of these viruses in farmed fish. Therefore, in this study, the pathogenicity of 10 isolates has been assessed with an in vivo trial. The investigation was conducted using the European sea bass, the first target fish species for the disease in the Mediterranean basin. Naive fish were challenged by immersion and clinical signs and mortality were recorded for 68 days; furthermore, samples collected at selected time points were analysed to evaluate the development of the infection. Finally, survivors were weighed to estimate the growth reduction. The statistically supported results obtained in this study demonstrated different pathogenicity patterns, underlined the potential risk represented by different strains in the transmission of the infection to highly susceptible species and highlighted the indirect damage caused by a clinical outbreak of VER/VNN.  相似文献   

13.
Abstract. The pathogenicity of the agent causing viral nervous necrosis (VNN) of striped jack, Pseudocaranx dentex (Bloch & Schneider), was examined in striped jack and other selected marine fish species. Fish were exposed to purified striped jack nervous necrosis virus (SJNNV) (0·1–100 ng ml−1) or homogenates of diseased striped jack larvae. Striped jack larvae (3·5 and 4·4 mm total length) were susceptible to the virus, but juveniles (78 mm) were not. The viral antigens were detected by indirect ELISA and the characteristic pathological changes, i.e. vacuolation in the retina and brain, were reproduced in the affected larvae. The infection was also established in healthy larvae by cohabitation with the diseased larvae. Larvae of red sea bream, Pagrus major Temminck & Schlegel, yellowtail, Seriola quinqueradiata Temminck & Schlegel, and goldstriped amberjack, Seriola lalandi Valenciennes, were not susceptible to SJNNV.  相似文献   

14.
鱼类神经坏死病毒研究进展与发展趋势   总被引:2,自引:2,他引:0       下载免费PDF全文
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。  相似文献   

15.
神经坏死病毒(Nervous necrosis virus)是导致多种海水鱼类神经性病害的致病原.发病及死亡的石斑鱼除了表现神经异常症状外,无明显的临床病症,体表及内脏组织也未发现明显病变及寄生虫感染.2003年4~8月,应用逆转录聚合酶链式反应(RT-PCR)技术从福建南部人工养殖的5种石斑鱼即紫石斑鱼(Epinephelus lanceolatus)、马拉巴石斑鱼(E. malabaricus)、青石斑鱼(E. awoara)、赤点石斑鱼(E. akaara)和云纹石斑鱼(E. moara)中检出5个神经坏死病毒分离株.检测了76份石斑鱼样品,这些石斑鱼NNV病毒的平均感染率约为90%.对这些病毒的RT-PCR产物421 bp核酸进行了测序和序列分析,其相同的序列超过99%.将这些序列与GenBank的石斑鱼(Epinephelus spp.)神经坏死病毒相关基因序列作比较,同源性在97%以上.对神经坏死病毒在石斑鱼体内的分布也进行了分析,在脑和眼组织的检出率最高,部分病鱼的肝、脾和肾组织也能检出病毒.结合流行病学特征,可确认神经坏死病毒为该传染病的主要致病原.RT-PCR方法是检测NNV等病原的一种理想的诊断方法.  相似文献   

16.
Mass mortalities of hatchery-reared juvenile groupers have occurred in southern Taiwan. The diseased fish swam in a darting, corkscrew fashion. Light microscopy revealed vacuolation in the brain tissue. Electron microscopy showed numerous non-enveloped, cytoplasmic viral particles (20–25 nm in diameter) in the brain cells, and many virions were enclosed in the membrane-bound organelles of the cells. Two structural proteins of the purified grouper virus, with molecular weights of 44 and 43 kDa, were revealed by SDS-PAGE. Moreover, the results of RT-PCR and nested PCR diagnosis using primers specific to the T2 and T4 target segments of striped jack nervous necrosis virus (SJNNV) RNA2 genes suggest that this virus is a fish nodavirus, and is designated as GNNV 9410 strain (grouper nervous necrosis virus strain 9410). This is the first case report of viral nervous necrosis among marine fish in Taiwan.  相似文献   

17.
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.  相似文献   

18.
19.
A betanodavirus associated with a massive mortality was isolated from larvae of tilapia, Oreochromis niloticus , maintained in fresh water at 30 °C . Histopathology revealed vacuolation of the nervous system, suggesting an infection by a betanodavirus. The virus was identified by indirect fluorescent antibody test in the SSN1 cell line and further characterized by sequencing of a PCR product. Sequencing of the T4 region of the coat protein gene indicated a phylogenetic clustering of this isolate within the red-spotted grouper nervous necrosis virus type. However, the tilapia isolate formed a unique branch distinct from other betanodavirus isolates. The disease was experimentally reproduced by bath infection of young tilapia at 30 °C. The reservoir of virus at the origin of the outbreak remains unidentified. To our knowledge, this is the first report of natural nodavirus infection in tilapia reared in fresh water.  相似文献   

20.
Betanodaviruses are the causative agents of the disease known as viral nervous necrosis (VNN) or viral encephalopathy and retinopathy (VER) in a variety of marine and freshwater fish species. The aim of this study was to demonstrate experimental infection of an isolate of betanodavirus (RGNNV genotype) in freshwater fish, Gambusia affinis, for elucidation of transmission mechanism and potential use as a laboratory model. Morbidity and mortality rate was significantly higher by injection route of infection as compared to immersion by bath and resembled the natural infection of juvenile marine fish. The fish in disease affected group showed severe neurological disorders accompanied by extensive vacuolar degeneration and mild to moderate neuronal necrosis of the brain in comparison to control. Amplification of ~?427 bp product in the variable region of the coat protein gene of betanodavirus was achieved by RT-PCR with 100% sequence homology to RGNNV genotype.  相似文献   

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