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.     

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.     

Cryopreservation of turbot Scophthalmus maximus (L.) sperm: fertilization and hatching rates

The present paper assesses the fertilization and hatching rates of an artificial fertilization series (n=1153) using fresh and cryopreserved sperm from 49 specimens of turbot Scophthalmus maximus (L.), carried out to confirm the results of a previous study, with the ultimate aim of transferring these cryo‐preservation techniques to commercial hatcheries. No significant differences were found between the fertilization rates of the two groups (fresh and cryopreserved sperm) when their respective fertility rates were >69.2%, which was the case in 75% of all fertilizations. Likewise, no significant differences in hatching rates were found. In order to use the cryopreserved sperm more efficiently, a key concern for commercial use of this technique, we also experimented with a lower sperm:diluent ratio (1:1) than used previously at our centre. We also compared the traditional 0.5‐mL straws with 2‐mL cryotubes able to contain a higher volume of sperm, finding no significant differences in the resulting fertilization and hatching rates. In conclusion, the use of cryopreservation for turbot sperm presents major advantages for broodstock management in commercial hatcheries.     

大菱鲆胚胎玻璃化方法研究

用大菱鲆(Scophthalmus maximus)神经期胚对6种抗冻剂的毒性进行检测,发现其毒性排列为1,2-丙二醇(PG)<甲醇(MeOH)<甘油(Gly)<二甲基甲酰胺(DMF)<乙二醇(EG)<二甲亚砜(DMSO).以DMF为主因子配制和筛选出11种玻璃化程度较好的玻璃化液,并用大菱鲆肌节期胚对其中5种玻璃化程度最好的玻璃化液进行检测,结果显示,A4(DMSO 20%+DMF 25%)、A7(DMSO 25%+DMF 20%)较适合于大菱鲆胚胎的平衡处理,利用A4和A7平衡处理神经期胚、肌节期胚、心跳期胚、出膜前期胚,结果显示大菱鲆神经胚和肌节胚对玻璃化液的适应能力较强.实验测试出不同时期胚胎在两种玻璃化液中的成活率,为大菱鲆各期胚胎在玻璃化液中的平衡处理提供了依据.利用大菱鲆肌节胚对不同的平衡步骤进行了筛选,发现五步法的平衡效果较好.     

Statistical properties and performance of pairwise relatedness estimators using turbot (Scophthalmus maximus L.) family data

The statistical properties and performance of four estimators of pairwise relatedness were evaluated in several scenarios using the microsatellite genotype data from a set of large known full‐sibships of turbot. All estimators showed a significant negative bias for the four kinships commonly used in these studies (unrelated: UR, half‐sibs, full‐sibs and parent–offspring), when allele frequencies of the reference population were estimated from the individuals analysed. When these frequencies were obtained from the base population from which all families proceeded, the bias was mostly corrected. The Wang (W) and Li (L) estimators were the least sensitive to this factor, while the Lynch and Ritland (L&R estimator) was the highest one. The error (mean around 0.130) was very similar in all scenarios for W, L and Queller and Goodnight (QG) estimators, while L&R was the highest error‐prone estimator. Parent–offspring kinship resulted in the lowest error, when using W, L and QG estimators, while UR resulted in the lowest error with the L&R estimator. Globally, W was the best‐performing estimator, although L&R could perform better in specific sampling scenarios. In summary, pairwise estimators represent useful tools for kinship classification in aquaculture broodstock management by applying appropriate thresholds depending on the goals of the analysis.     

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。     

Experimental transmission of encephalopathy and retinopathy induced by nodavirus to sea bream, Sparus aurata L., using different infection models

The transmission of viral encephalopathy and retinopathy (VER) was investigated in juvenile sea bream, Sparus aurata L. Two different infection routes [intraperitoneal (i.p.) and intramuscular injection (i.m.)] were tested at two different temperatures (20 and 26 °C) using sea bream of mean weight 0.7, 2 and 4 g, as well as an immersion challenge performed at 26 °C with sea bream of 0.7 g. Successful transmission of the disease was only achieved by i.m. injection. Mortalities of 100% occurred in sea bream of 0.7 g at day 15 post-infection and 47% in sea bream of both 2 and 4 g at day 30 post-infection in all the experimental infections at 26 °C. No mortalities were ever observed with infections at 20 °C. When mortalities were observed, the virus was detected by immunoperoxidase staining in the SSN-1 cell line inoculated with tissues from infected fish. Histological examination of both normal and infected fish showed a vacuolization in the bipolar and granular layers of the retina of the infected sea bream. This is the first experimentally induced transmission of VER in sea bream. Differences were observed at the time of disease onset depending on water temperature, the route of infection and the age of the juvenile fish.     

养殖大菱鲆蟹栖异阿脑虫感染及其危害

从养殖大菱鲆（Scophthalmus maximus）患病鱼体中分离出一种寄生性纤毛虫，通过活体观察、碳酸银法染色和扫描电镜对其形态学进行特征分析。经鉴定，该纤毛虫为蟹栖异阿脑虫（Mesanophrys carcini Groliere ＆ Leglise，1977），隶属于寡膜纲、盾纤目、嗜污科、异阿脑虫属。患病鱼常表现为静伏池底，体色变暗，活力弱，摄食差，生长减慢，并发生急、慢性死亡。最明显的特征是病灶部位变白，略有浮肿，触摸柔软。流行病学特征显示，在大菱鲆的苗期、养成期、亲鱼培育期均可发生纤毛虫感染，幼鱼期发生率较高，发病水温14-20℃，盐度12～40。组织病理学观察分析表明，病灶组织处以盾纤虫为主要入侵微生物，常伴有细菌存在，但没有发现病毒性细胞病变和霉菌等其他寄生生物。     

Experimental infection of turbot, Scophthalmus maximus (L.), by Moritella viscosa, vaccination effort and vaccine-induced side-effects

Moritella viscosa is the causative agent of winter ulcers in farmed salmonids and Atlantic cod in countries around the North Atlantic. The bacterium has also been isolated from various marine fish species. Bacterial diseases have been a limiting factor in farming of turbot, but M. viscosa has not so far been isolated. In this study, turbot was shown to be sensitive to M. viscosa infection in experimental challenges. Pathological changes in infected turbot were comparable with those previously described for winter ulcers in salmon. A multivalent commercial salmon vaccine, containing M. viscosa as one of five antigens and a mineral oil adjuvant, did not protect turbot against challenge 13 weeks post-vaccination. Weight gain of vaccinated turbot compared with controls was not reduced 7 weeks post-vaccination. Vaccination did not induce a specific anti-M. viscosa response, while elevated anti-M. viscosa antibody levels were detected both in vaccinated and unvaccinated fish 5 weeks post-challenge. The vaccine did, however, induce an antibody response against Aeromonas salmonicida, another vaccine component. Minor intra-abdominal adhesions were detected in vaccinated fish and fish injected with a mineral oil adjuvant. The measurement of various innate humoral immune parameters did not reveal significant differences between vaccinated and control groups.     

Edwardsiellosis in farmed turbot,Scophthalmus maximus (L.), associated with an unusual variant of Edwardsiella tarda: a clinical,aetiological and histopathological study

During 2005 and 2010, a survey of edwardsiellosis on eight turbot, Scophthalmus maximus (L.), farms was conducted in China. This report presents the detailed results of the study on this disease. Diseased turbot displayed two distinct types of gross signs: black discoloration of the dorsal skin on the posterior portion of the body; and red cutaneous foci on the ventral side. Internally, the most pronounced clinical signs in all fish examined were enlarged kidneys. The causal agent of the disease was finally proved to be one species of bacterium that was identified as Edwardsiella tarda by physiological and biochemical tests, API 32E and 16S ribosomal RNA sequence analysis. It is noteworthy that unlike the commonly described E. tarda strains, the isolates in this study were non‐motile strains without flagella. A histopathological study revealed that E. tarda infection was systemic in turbot and that kidney showed the most significant pathological changes, including acute focal necrosis, an influx of macrophages and formation of granuloma. The most common histopathological characteristics of this disease are the proliferation of macrophage in various organs and formation of granuloma. In addition, this article also gave background information on the disease and presented the results of virulence tests with the E. tarda strain identified in this study.     

Isolation of a highly pathogenic Vibrio pelagius strain associated with mass mortalities of turbot,Scophthalmus maximus (L.), larvae

A bacterial strain, characterized as Vibrio pelagius (Hq 222), was isolated from a turbot, Scophthalmus maximus (L.), larvae mass mortality in a commercial fish farm in Spain. Turbot larvae, post-larvae (0.2 g) and juveniles (5 and 15 g) were experimentally infected. The bacterium appeared to be very virulent for larvae and post-larvae, LD50 being < 5 bacteria mL(-1) for larvae 1 week post-infection and 3.9 x 10(5) bacteria mL(-1) in post-larvae at day 12 post-infection. The bacterial strain was recovered in pure culture from the internal organs of infected fish. Histological lesions in post-larvae exhibited swelling and necrosis of gill secondary lamellae, sloughing of intestinal mucosa and necrosis of haematopoietic tissue in the kidney. Vibrio pelagius (Hq 222) was able to grow in sterile sea water when incubated at room temperature or at 15 degrees C. Vibrio pelagius (Hq 222) was more adherent to the turbot cell lines TV-1 and TF than Escherichia coli. In both cell lines, the number of adhered bacteria increased with incubation time.     

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)。     

Effects of dietary water content on meal size, daily food intake, digestion and growth in turbot, Scophthalmus maximus (L.)

When fed once daily with wet squid, turbot (30–50 g) accustomed to dry pellets require many days to increase intake to meet their feed requirement (≈ 10 mg dry matter g⁻¹ bw meal⁻¹). Adaptation takes 1–2 days if several daily feedings are given. With dried squid, they ingest about 20% of the wet squid bulk because the stomach contents expand when moisturised. In contrast, turbot eat enough wet squid to fill most of the available stomach volume (≈ 7.6 mL 100 g⁻¹ bw). When presented in gelatine capsules, food water content is masked and does not affect the volume ingested. Moistening the contents shortens the delay before gastric emptying starts to one-third (0.6 h) compared with dry food (1.9 h). Daily dry-matter intake increased when dry contents were moistened but only if two or more meals were offered per day. Turbot adapt their digestion to supply water for dry diets but this may add extra metabolic costs. When offered 20 mg dry matter g bw⁻¹ day⁻¹, divided into four equal meals, turbot grew faster and more efficiently with moist than with dry squid. Protein, energy and dry-matter digestibilities were also enhanced. The increased daily protein absorption did not increase ammonia release, indicating that the extra protein was used for somatic growth.     

Physiological and morphological effects of severe hypoxia,hypoxia and hyperoxia in juvenile turbot (Scophthalmus maximus L.)

We studied the physiological status of juvenile turbot (Scophthalmus maximus L.) under severe hypoxia (1 and 2 mg/L dissolved oxygen, DO), hypoxia (3 and 5 mg/L DO), hyperoxia (11 and 14 mg/L DO) and normoxia (7 mg/L DO, control) conditions. The respiratory rates, haematology parameters, acid–base balance status and gill structure were analysed to find the effects of different DO concentration on turbot. Fish mortality was only observed under severe hypoxia conditions. Severe hypoxia caused an increase in respiratory rates and red blood cell counts, as well as an increase in haemoglobin and haematocrit levels in the fish. In fish exposed to hypoxia conditions, the respiratory rate increased overall as the DO concentration decreased. Lower pCO₂ and HCO₃^? levels led to a high blood pH, while the pO₂ remained stable. In hyperoxia groups, respiratory rate decreased as the DO concentration increased. The levels of pCO₂ and HCO₃^? significantly increased (P < 0.05), while the pO₂ level and blood pH did not change obviously. The gill structure was damaged after prolonged exposure to hyperoxia, but no obvious damage was found in hypoxia groups. The fish that survived the hypoxia or hyperoxia treatment were able to restore the structural integrity of the gills after 14 days' recovery. The results suggest that juvenile turbots can tolerate a wide range of DO concentrations. However, even mild hyperoxia condition (11 mg/L DO), which is widely used in fish culture, has adverse effects on juvenile turbot physiology.     

大菱鲆体重性状不同生长期的遗传分析

采用条件数量性状加性-显性遗传模型和统计分析方法, 对大菱鲆(Scophthalmus maximus)体重性状不同生长期进行遗传分析; 基于von Bertalanffy模型, 对大菱鲆6种交配组合的生长曲线划分为缓慢生长期、快速生长前期、快速生长后期和渐进期, 并与各生长期所对应的遗传分析结果进行比对。结果表明, 6种交配组合的缓慢生长期、快速生长前期和快速生长后期基本上是一致的。在缓慢生长期, 检测到新的条件加性方差和条件显性方差, 其相应的条件加性方差分量比率均未达到显著水平, 相应的条件显性方差分量比率均达到显著(P<0.05)或极显著水平(P<0.01); 在快速生长前期, 交互检测到达到极其显著水平的条件加性方差和条件显性方差, 其相应的条件加性方差分量比率和条件显性方差分量比率分别在15~18月龄和12~15月龄达到极显著水平(P<0.01); 在快速生长后期, 同时检测到达到极其显著水平的条件加性方差和条件显性方差, 其相应的条件加性方差分量比率和条件显性方差分量比率均达到极其显著水平(P<0.01), 但同期的条件显性方差分量比率高于条件加性方差分量比率。本研究通过探讨大菱鲆体重性状在特定生长发育期的遗传机理, 深入了解复杂数量性状时空动态变化的遗传生态学机制及其遗传规律, 旨在为提高大菱鲆育种的预见性和选择效率提供理论参考。

     

大菱鲆AFLP分析体系的建立

本研究构建了大菱鲆扩增片段长度多态性(AFLP)分析体系,对DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤进行了分析。结果表明,用EcoRI/MseI双酶切、核心序列加3个选择性碱基的选择性扩增引物、1∶6(w∶w)的EcoRI端与MseI端选择性扩增引物比和20μl选择性扩增体积,可得到十分稳定的结果。所得产物经电泳和银染后可获得条带清晰、背景干扰小的图像。该体系的构建为AFLP技术在大菱鲆相关研究中的应用奠定了基础。