Pathology of Edwardsiella tarda infection in turbot, Scophthalmus maximus (L.)

Macroscopic and histopathological changes in cultured turbot, Scophthalmus maximus (L.), in Spain caused by infection with Edwardsiella tarda are described. Eye tumefaction, inflammation, haemorrhages, ascites and the presence of a purulent fluid were the main macroscopic lesions observed. Histopathological lesions were found in the kidney, spleen and liver. In the kidney and spleen these were characterized by a severe apostematous inflammatory reaction, with a large number of abscesses. The liver was affected to a lesser degree and only some phagocytes loaded with bacteria were observed. Ultrastructural observations indicated that macrophages were the main cell type implicated in the inflammatory response. Most of the bacteria observed within the phagocyte cytoplasm showed no degenerative changes and some were dividing. Degenerative changes observed in macrophages indicate their failure in preventing the infection.     

Systemic infection with Uronema-like ciliates in farmed turbot, Scophthalmus maximus (L.)

Histophagous ciliates caused high mortality among turbot in a commercial fish farm in southern Norway. The ciliates spread systemically in fry (< 0.3 g). In the early stages of infection the ciliates were found in connective tissue in skin and fins, as well as in nervous tissue. In terminal stages the whole organism was infected. In large turbot (500–1000 g), the ciliates were predominantly found in the central nervous system, causing liquefaction of the nervous tissue. The ciliates were not identified to species but resembled species in the genus Uronema (Scuticociliatida).     

An amoeba associated with gill disease in turbot,Scophthalmus maximus (L.)

An amoeba associated with a severe gill disease affecting farmed turbot, characterized by pronounced hyperplasia of the branchial epithelia and accumulation of mucus between the secondary lamellae, is described for the first time. The amoeba was isolated, cloned and cultured on Bactoagar. Light microscopy showed that trophozoites had a mean length of 12 μm (range 8–20 μm), with a nucleus 3–5 μm in diameter containing a large central nucleolus 1–3 μm in diameter. Transmission electron microscopy showed that the mitochondria, Golgi bodies, rough endoplasmic reticulum, microfilaments, cytoplasmic microtubules and glycocalyx were similar to those previously reported for the genus Platyamoeba. These and other findings strongly suggest that the amoeba isolated is of the genus Platyamoeba, while its morphometric characteristics suggest that it may represent a new species. This is the first report of a species of the genus Platyamoeba in association with gill disease in turbot.     

Glucose tolerance in turbot, Scophthalmus maximus (L.)

Eighty turbot, Scophthalmus maximus (L.), (average weight 61 g) were injected intraperitoneally with exactly 1 g glucose per kg body weight. There was a peak in plasma glucose 3 h post injection. Thereafter a gradual decrease to basal levels was seen within 24 h. Plasma triacylglycerol concentrations showed a rapid decline during the first 24 h, and thereafter stable values. Blood haematocrit values decreased from 20% before injection to 16% 72 h after injection. Liver glycogen concentrations showed an initial decrease from 8 to 5 g 100 g^?1 (w.w) during the first 12 h, and thereafter stable values, while muscle glycogen concentrations increased during the first 12 h, and thereafter showed a gradual decline until 72 h. This response was most probably caused by secondary changes upon handling in combination with the direct response to a glucose load. Thus turbot was able to restore alterations in carbohydrate metabolism efficiently within 24 h.     

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.     

Characterization of nodavirus and viral encephalopathy and retinopathy in farmed turbot, Scophthalmus maximus (L.)

An outbreak of nodavirus infection in turbot larvae is described with respect to histopathology, immunohistochemistry, cell culture cultivation, RT-PCR amplification and sequence analysis of the capsid protein gene RNA2. Affected turbot developed classical signs of viral encephalopathy and retinopathy (VER) with abnormal swimming behaviour and high mortality levels. In the acute stage of infection, light microscopy revealed vacuolation of the central nervous system (CNS), with positive immunohistochemical staining for nodavirus. Later in the infection, CNS lesions appeared more chronic and contained clusters of cells immunopositive for nodavirus. Bacterial overgrowth in the intestines of the fish may have provoked or influenced the course of the nodavirus infection. We were unable to propagate the virus in cell culture. While RT-PCR using primers designed to detect Atlantic halibut nodavirus gave negative results, further testing with primers complementary to a more conserved region of RNA2 resulted in amplification of a product of the expected size. The entire RNA2 segment was cloned and sequenced. Sequence alignment showed that the turbot nodavirus (TNV) was different from previously described fish nodaviruses. In addition, phylogenetic analysis based on an 823 nt region of the sequence indicated that TNV clustered outside the four established fish nodavirus genotypes, suggesting a fifth genotype within the betanodaviruses.     

Studies on Herpesvirus scophthalmi infection of turbot Scophthalmus maximus (L.) ultrastructural observations

Abstract. Recent heavy mortalities amongst O+ group turbot at a fish farm were found to be associated with a herpes-type viral infection of the epithelia of the skin and gills. The morphology of the virus is described with ultrastructural observations on its morphogenesis and release from infected cells.     

Metabolizable energy from dietary carbohydrates in turbot, Scophthalmus maximus (L.)

Abstract. In a feeding experiment, the metabolizability of dietary carbohydrates in diets for turbot, Scophthalmus maximus (L.), was estimated and expressed as percentage of the metabolizability of dietary lipids. Four isoenergetic and isonitrogenous dietary treatments were applied with varying percentages of non-protein energy presented as starch. In order to obtain results allowing quantitative interpretation, an experimental design was used in which effects are reflected in varying body fat depositions only. Two methods were used simultaneously to overcome practical problems associated with differences in energy density between carbohydrates and lipids. The conventional method was based on equal feeding rates combined with the inclusion of inert filler in the diets, whereas the alternative method applied different dietary nutrient percentages in combination with different feeding rates. Although similar results were obtained, the alternative method was more convenient to use than the conventional method. Because of the experimental strategy applied, no significant effects on growth and protein utilization were obtained, but lipid deposition rates were significantly reduced at increased percentages of non-protein energy presented as starch. The metabolizable fraction of the gross dietary carbohydrate energy was estimated as 67% of that of dietary lipids. This reduction does not indicate that turbot have any systematic deficiency in metabolizing dietary carbohydrates.     

一种吻蛭类大鱼蛭在大菱鲆鱼体上的感染

在山东省威海崮山育苗场发现大菱鲆成鱼及幼鱼鱼体感染寄生虫，受感染的鱼由于处于感染初期，并经过及时的处理，所占比例较低。病鱼症状为鱼体体色变黑、瘦弱、离群、不摄食甚至死亡。寄生虫寄生于大菱鲆的体表及鳃部，虫体黑褐色，全长40～50mm。经鉴定，该寄生虫属于环节动物门(Annelida)，蛭纲(Hirudinea)，吻蛭目(Rhynochobdellida Blanchard)，鱼蛭科(Piscicolidae Johnson)，鱼蛭属(Piscicola Blainville)，大鱼蛭(Pisciola magna Yang)。     

Dietary protein requirement for young turbot (Scophthalmus maximus L.)

This study was conducted to determine the optimum dietary protein level for young (an initial weight of 89 g) turbot, Scophthalmus maximus L. Duplicate groups of the fish were fed the five isoenergetic diets containing the various protein levels ranging from 290 to 570 g kg^?1 diet for 45 days. Survival was not affected by dietary protein level. Weight gain and feed efficiency were improved with dietary protein level up to 490 g kg^?1 diet. Dietary protein requirement of young turbot using the broken‐line model was estimated to be 494 g kg^?1 diet based on weight gain response. Protein efficiency ratio was not influenced by dietary protein level. The highest protein retention was obtained from the fish fed the 490 g protein kg^?1 diet. Proximate composition of the fish was not significantly affected by dietary protein level. In considering these results, it was concluded that the 494 g protein kg^?1 diet with 100 g lipid kg^?1 diet (15 MJ kg^?1 diet) provided optimal growth of young turbot under these experimental conditions.     

Studies on Herpesvirus scophthalmi infection of turbot Scophthalmus maximus (L.): Histopathological observations

Abstract. The histological features of the disease caused by infection of turbot by Herpesvirus scophthalmi are described. Infection of epidermal cells of skin and gill resulted in the formation of giant cells showing a variety of nuclear and cytoplasmic changes associated with the replication of the virus. The effects of the condition together with possible control measures are discussed.     

Ulcerative skin and fin lesions in reared turbot, Scophthalmus maximus (L.)

Abstract. In this report, the authors describe the evolution of an ulcerative disease that occurred during July and August 1986 in cultured juvenile turbot, Scophthalmus maximus (L.), in northwestern Spain. The daily fish mortality ranged from less than 1 to 7·5%, and the total of cumulative losses were 23·2%. The start of the epizootic was closely associated with a sudden increase of water temperature to values higher than 20·C. Peaks of mortality were observed during the development of the disease. Affected turbot showed erratic swimming, respiratory distress and inappetence. The lesions began as white discoloured foci on the skin, operculum, and base of dorsal and caudal fins which could suffer a total destruction of the interradial membrane. In an advanced stage of this condition, the epithelial surface of the centre of the lesions sloughed off and shallow and haemorrhagic ulcers were produced. Three distinct areas were observed in the ulcerative lesions with a clear predominance of different microorganisms: (1) a peripheric pale zone invaded by a ciliate protozoan resembling Cryptocaryon spp.; (2) an intermediate greyish-dark area containing large numbers of fastidious myxobacteria which failed to grow in culture media; and (3) a central haemorrhagic zone where we have isolated two halophilic, urease positive Vibrio strains (closely related to V. carchariae ). No microbial involvement was detected in the internal organs of affected fish. The virulence assays conducted with the Vibrio strains failed to reproduce the signs of the disease, either by intraperitoneal inoculation or by water challenge after skin abrasion. The use of alternate baths of formalin/malachite green and nitrofurazone resulted in an effective therapy to control the mortalities.     

Haemorrhagic areas in the mouth of farmed turbot, Scophthalmus maximus (L.)

Abstract. An epizootic in juvenile turbol reared on a farm located in The Ria de Vigo, Galicia, Northwest Spain, is described. The diseased turbot did not display unusual swimming behavior; the external signs of the disease were abdominal distension and haemorrbagic areas in the mouth. Internal examination of the fist showed an accumulation of reddish fluid in the peritoneal cavity. Microbiological analysis of the diseased fish revealed the presence, in pure culture in all the organs and lesions examined, of a bacterium which was characterized biochemically as Vibrio splendidus biotype I. The virulence tests showed that the V. splendidus biotype I isolate was pathogenic for rainbow trout (LD₅₀: 2.2 × 10⁴) and also for turbot (LD₅₀: 1.2 × 10⁴). The treatment of the fish using flumequine incorporated into the feed was effective in offsetting the mortality rate.     

Reassortant betanodavirus infection in turbot (Scophthalmus maximus)

In this study, the susceptibility of turbot juveniles to two betanodavirus strains was assessed, a RGNNV/SJNNV reassortant (Ss160.03) and a SJNNV strain. The reassortant isolate exhibits a slightly modified SJNNV CP, with two amino acid substitutions in the C‐terminal domain (positions 247 and 270). To analyse the role of these residues as virulence and host determinants in turbot, three recombinant strains (rSs160.03₂₄₇, rSs160.03₂₇₀, rSs160.03_247+270) harbouring site‐specific mutations in the CP sequence were also tested in experimental trials. Moderate mortalities (up to 50%) were recorded at 18 °C in the fish challenged with the Ss160.03 strain, whereas low mortalities (17%) were observed in the group challenged with the SJNNV strain. A slight decrease (around 10%) was observed in the mortalities caused by the mutants rSs160.03₂₄₇ and rSs160.03₂₇₀, whilst the mutation of both positions reduced mortality by more than half of that observed in fish challenged with the wild strain. These results are confirmed by the replication in brain tissues, because whereas the wild strain was detected from 5 to 30 dpi and reached the highest viral load, the recombinant virus harbouring both mutations was not detected in the brain until 20 dpi and with a moderate viral load.     

Larval growth of turbot, Scophthalmus maximus (L.) produced with fresh and cryopreserved sperm

The present paper assesses the fertilization and hatching rates, as well as the growth, of larvae obtained from four artificial fertilizations (AF) using fresh and cryopreserved sperm of the turbot Scophthalmus maximus (L.). Larvae growth in both sperm groups, measured in terms of length and weight at culture days 0, 7, 14 and 31, are compared, as well as their growth rates. The two groups' fertilization and hatching rates were not significantly different. Likewise, no significant differences in length and wet weight of 7‐ and 14‐day‐old larvae were found using fresh and cryopreserved sperm; however, significant differences were found in 31‐day‐old larvae, which were more attributable to the variability inherent in larval turbot culture, and to variability in the reproductive specimens used in our study, than to the type of sperm employed. These results indicate that the type of sperm used in artificial fertilization, i.e. fresh or cryopreserved, is not a determining factor, either for fertilization and hatching, or for subsequent larval development. Our results also confirm once again the high quality of cryopreserved turbot sperm, and its usefulness in commercial hatcheries.     

大菱鲆种质资源研究与开发

综合介绍了大菱鲆种质资源的分类、自然分布和生物学特性。从生化和分子生物学方面阐述了大菱鲆自然群体遗传结构及国内外研究现状。详述了大菱鲆野生资源的利用和养殖开发现状．特别是我国对引进的大菱鲆种质资源的养殖开发。指明了我国目前在引种及养殖方面存在的不足．提出了建立全国性大菱鲆良种引育中心,以保证我国大菱鲆养殖业的持续、稳定、健康发展的建议。     

Protein kinase C in the spleen of the turbot (Scophthalmus maximus L.)

PKC activity was detected in spleen extracts from the turbot, Scophthalmus maximus, a teleost flatfish that is farmed commercially in several countries, in assays with the substrate EGF- R651–658 as phosphate acceptor. The activity was purified about 700-fold by a three-step chromatographic procedure (DEAE-cellulose, phenyl-Sepharose and threonine-Sepharose). Maximal activity was obtained in the presence of the typical PKC cofactors Ca2+ (0.1 mM) PtdS (20 g ml–1) and either DAG (2 g ml–1) or PMA (2 g ml–1). Activity was dose-dependently inhibited by H7 and by the PKC-specific inhibitors PKC19–36 and N-myristoylated PKC19–31. The rate of phosphorylation was highest with the PKC-specific substrate MARCKS161–175. In immunoblotting, MC5 (a mouse monoclonal antibody raised against bovine PKC) recognized bands of 80 and 100 kDa. Immunoblotting with antibodies raised against mouse PKC isozymes (, , , , , , and ) indicated the presence of all these isozymes in turbot spleen.     

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。