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.     

Dietary nucleotides improve the growth performance,antioxidative capacity and intestinal morphology of turbot (Scophthalmus maximus)

A growth trial was conducted to evaluate the effects and safety of nucleotides in low fish meal diets on the growth performance, antioxidative capacity and intestinal morphology of turbot (Scophthalmus maximus). High fish meal control diet was formulated with 500 g kg^?1 fish meal. Seven levels (0.075, 0.15, 0.225, 0.300, 1.5 and 3.0 g kg^?1, respectively) of nucleotides were added to a low fish meal basal diet, which was formulated with 400 g kg^?1 fish meal. The eight experimental diets were fed to groups of juvenile turbot (initial weight: 6.0 ± 0.03 g) for 60 days. Results showed that compared with high fish meal control diet, low fish meal basal diet treatment had lower total antioxidative capacity (T‐AOC), glutathione peroxidase activity, fold height of proximal and distal intestine, enterocyte height of all evaluated enteric section and microvillus height of mid‐intestine and distal intestine (P < 0.05). However, supplemented nucleotides in diets could significantly improve growth (specific growth rate, SGR), feed utilization, antioxidative capacity and intestinal morphology of turbot (P < 0.05). Broken‐line regression analysis of SGR and T‐AOC showed that the optimal supplemental levels of dietary nucleotide for juvenile turbot were 0.366 and 0.188 g kg^?1, respectively. In summary, 0.300 g kg^?1 of dietary nucleotides was helpful in improving growth, feed utilization, antioxidative capacity and intestinal morphology of turbot fed with low fish meal diet. Excessive dietary nucleotides (3.0 g kg^?1) might cause oxidative stress and morphological damage in intestine and then reduce the growth of turbot.     

Influence of several non‐nutrient additives on nonspecific immunity and growth of juvenile turbot,Scophthalmus maximus L.

The effects of three non‐nutrient additives on nonspecific immunity and growth of juvenile turbot (Scophthalmus maximus L.) were studied in this feeding experiment. The five treatments are basal diet alone, basal diets containing three different additives [0.4 g kg^?1 of xylo‐oligosaccharides (XOS), 1.3 g kg ^?1 of yeast cell wall and 0.8 g kg ^?1 of bile acids] individually or in combination. Two hundred and twenty‐five turbots (average initial weight 151.3 ± 11.3 g) were randomly allotted in five treatments with three replicates within each treatment in a 72‐day period. Comparing with basal diet group, activities of C3, C4, phagocyte, lysozyme, specific growth rate and feed conversion rate in yeast cell wall, XOS and the combined groups was enhanced significantly (P < 0.05); however, these parameters in bile acid groups were increased slightly (P > 0.05) except for phagocyte (P < 0.05); superoxide dismutase activity in additive groups was not significantly increased (P > 0.05) except for the combined group (P < 0.05). In conclusion, supplementation of yeast cell wall and XOS enhanced the nonspecific immunity of juvenile turbot. Synergistic or additive effect of the three additives was not observed.     

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.     

Alleviation effect of dietary cerium and its complex with chitosan oligosaccharide on cadmium accumulation in juvenile turbot,Scophthalmus maximus L., under cadmium stress

An 8 weeks feeding trial was conducted to investigate the effect of dietary cerium (Ce) and its complex with chitosan oligosaccharide (COS‐Ce) on growth performance and cadmium (Cd) accumulation of turbot, Scophthalmus maximus L. under Cd stress. The basal diet (Diet 0) was formulated without Cd and cerium as the control. Seven other experimental diets (Diets 1–7) were formulated with supplementation of 50 mg Cd²⁺/kg feed, 50 mg Cd²⁺/kg and 50 mg Ce³⁺/kg feed, 50 mg Cd²⁺/kg and 100 mg Ce³⁺/kg feed, 50 mg Cd²⁺/kg and 200 mg Ce³⁺/kg feed, 50 mg Cd²⁺/kg and 50 mg COS‐Ce/kg feed, 50 mg Cd²⁺/kg and 100 mg COS‐Ce/kg feed, and 50 mg Cd²⁺/kg +200 mg COS‐Ce/kg feed. Results of the present study showed that, compared with the control group, the condition factor in fish fed the diet with 50 mg Cd²⁺/kg feed (Diet 1) was significantly lower, whereas the Cd concentration in liver and kidney of fish fed the diet with 50 mg/kg Cd²⁺ (Diet 1) was significantly higher (P < 0.05). The high Cd accumulation of fish fed diets with sole 50 mg/kg Cd (Diet 1) could be significantly decreased by 51.72% after supplementation of 200 mg COS‐Ce/kg in the diet (Diet 7). These results suggested that elevated COS‐Ce could effectively protect against dietary Cd accumulation in turbot.     

温度对大菱鲆幼鱼生长、成活率和体内蛋白酶活性的影响

通过温度的变化研究不同温度对大菱鲆幼鱼生长、成活率和消化器官内蛋白酶活性的影响,结果表明,A在温度达到0℃时,幼鱼成活率为30%;4℃以上至20℃时成活率为100%,22~24℃为80%;26℃为40%;当水温达到28℃时,幼鱼成活率为0(20d)。养殖水温在8℃以下时,幼鱼基本不摄食,体重没有增长。12~16℃时,其生长速度随温度的升高而加快,当养殖水温达到20℃时,其生长速度与16℃条件下的生长速度相比较开始下降,达到24℃时,幼鱼生长速度已经明显下降,与12℃条件下的体重增长速度相接近。在8℃之前,幼鱼胃、肠和肝脏等消化器官中的蛋白酶活力单位含量较低。随温度的增加,酶活力单位显著增加,其中胃中的消化酶活力增加最快。     

Effects of silymarin on growth performance,antioxidant capacity and immune response in turbot (Scophthalmus maximus L.)

A 9‐week feeding trial was conducted using triplicate groups of turbot (6.50 ± 0.01 g) to explore the potential effects of silymarin. Three concentrations of silymarin (100, 200, and 400 mg/kg) were added to the plant protein‐based diet. Fish were randomly distributed into fiberglass tanks (30 fish per tank). The results showed that adding 100 mg/kg silymarin significantly improved the growth performance, with no effects on feed utilization. The antioxidant capacity in the liver was significantly improved in the 100 mg/kg and 200 mg/kg silymarin groups by not only inducing the activities of superoxide dismutase (SOD) and catalase but also increasing the messager RNA (mRNA) expression levels of SOD, glutathione peroxidase, and peroxiredoxin 6. Meanwhile, supplying 100 and 200 mg/kg of silymarin enhanced the heights of villi and enterocytes. Silymarin supplementation reduced the mRNA expression of interleukin‐8 and tumor necrosis factor‐α but induced the expression of transforming growth factor‐β (TGF‐β) in the intestine. These results indicated that silymarin was a potential nutraceutical that could enhance the growth performance and health status of turbot fed in a high plant protein diet. Adding 100 mg/kg silymarin to the plant protein diet achieved optimal performance in turbot.     

Effect of feed restriction on the growth performance of turbot (Scophthalmus maximus L.) juveniles under commercial rearing conditions

Two trials were performed to study the effect of periodic feed deprivation (trial 1) or feed restriction, followed by satiation feeding (trial 2) on the growth performance of turbot juveniles under commercial rearing conditions. In trial 1, duplicate groups of 350 fish with an initial weight of 62 g were fed a commercial diet to apparent visual satiation for 7, 6, 5 or 4 days a week for 83 days. At the end of the trial, fish weight was directly related to the number of feeding days but feed efficiency and protein efficiency ratio were not affected by treatments. At the end of the trial, there were no differences in whole‐body composition among groups. In trial 2, duplicate groups of 500 fish with an initial weight of 33 g were fed a commercial diet to satiation (100%) or feed restricted to 90%, 80% and 70% of satiation for 90 days. Thereafter, all groups were fed to satiation for 34 days. During the feed restriction period, growth was directly related to feed intake, while during the satiation feeding period, it was inversely related to the previous feeding level. At the end of the trial, the final weight was not different among groups. At the end of the feed restriction period, whole‐body lipid content showed a trend to decrease with an increase in the feed restriction level. The results of this study indicate that under practical conditions, turbot juveniles should be fed daily as even cycles of short periods of feed deprivation negatively affect growth, while not improving feed efficiency. On the contrary, even after a relatively long feed restriction period, fish shows compensatory growth, and this may be used as a feed management strategy for controlling fish production in commercial farms.     

Growth and survival of young turbot (Scophthalmus maximus L.) produced with cryopreserved sperm

This study assessed the growth and survival over a year of two groups of 4‐month‐old turbot Scophthalmus maximus (L.) derived from artificial fertilization with fresh (FG) and cryopreserved sperm (CG). Growth in both groups, measured monthly in terms of length and weight, were compared. Survival was also recorded. No significant differences were found when we compared weight and length data in both groups. Growth rates were similar between FG and CG young turbot during 1 year. Likewise, the same survival rate (92.2%) was found in both groups. Our results show the good survival and growth of young turbot obtained from cryopreserved sperm, and confirm the cryopreservation technique as a useful tool for raising turbot for commercial purposes.     

Growth, feed utilization and growth heterogeneity in juvenile turbot Scophthalmus maximus (Rafinesque) under different photoperiod regimes

Juvenile turbot (45 g, SE = 1.3) were reared under three photoperiods, 08L:16D, 12L:12D and 20L:04D at slightly elevated ambient temperature for Ireland. Over the 297‐day experimental period, the overall growth rate of the 12L:12D (0.82% d^?1) treatment was higher than for both 08L:16D (0.80% d^?1) and 20L:04D (0.77% d^?1). Overall relative feed intake (FI = % consumption*day^?1) was higher for the 20L:04D (FI = 0.81% d^?1, SE = 0.06) treatment than for the 08L:16D (0.63% d^?1, 0.04) and 12L:12D (0.64% d^?1 0.04) treatments, whereas feed conversion efficiency (FCE = weight gain* consumption^?1) was lower in the 20L:04D (FCE = 0.67, SE = 0.08) group when compared with the 08L:16D (0.88, 0.06) and 12L:12D (0.88, 0.06) treatments. Present results show that the long‐term extended fixed photoperiod may act as an irritant, inducing stress, suppressing growth and reducing feed utilization. It is hypothesized that the progression of size‐dependent hierarchies over time can be divided into two distinct phases herein referred to as ‘hierarchy resolution’ and ‘hierarchy stabilization’ phases (or phases 1 and 2) characterized by increasing and decreasing growth heterogeneity respectively. Growth heterogeneity is measured as coefficient of variation of weight and rank correlation of initial weight of a phase and corresponding growth rate.     

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.     

Estimation of genetic parameters for early growth trait of turbot (Scophthalmus maximus L.) using molecular relatedness

In this study, a data set of 1140 progenies from 19 families that were composed of 13 half‐sib and six full‐sib families was used in the estimation of the genetic parameters of weight for juvenile turbot at 100 days post hatch. Sixty progenies were randomly selected from each family, and 20 of these individuals were genotyped at 12 microsatellite loci. Two sorts of relatedness, including pedigree relatedness (PR) from a complete pedigree and average molecular relatedness (AMR) from molecular markers, were compared to explore the feasibility of AMR in selective breeding when only parents were known. Two different animal models, Model 1 and Model 2 which were without and with the addition of maternal and common environmental effects, respectively, were used to estimate the variance component and breeding values for weight using restricted maximum likelihood (REML) method. Thereafter, cross‐validation was applied to investigate predictive ability of model and accuracy of breeding values. Pearson correlation analysis showed that AMR was highly correlated (0.91) with PR. According to conditional Akaike Information Criterion, the best models for AMR and PR methods were Model 2 and Model 1 respectively. Heritability estimates from AMR and PR methods based on their best models were 0.19 (±0.06) and 0.66 (±0.17). Cross‐validation showed that AMR was comparable to PR in terms of predictive abilities and accuracies of breeding values based on the same model. This study therefore suggests that the AMR can be applied as alternative of PR when only parents were known.     

不同水循环率对大菱鲆生长和水质的影响研究

研究了封闭循环水养殖系统中不同水循环率对大菱鲆(Scophthalmus maximus)生长和水质变化的影响。试验设置4个水循环率梯度12,24,36,48次/d,大菱鲆初始平均体重为200.36 g。经43 d养殖,12次/d组大菱鲆最终平均体重为277.98 g/尾,而48次/d组达到了296.24 g/尾;12次/d组水体氨氮(TAN)和亚硝酸盐氮(NO2--N)浓度分别为0.41~1.50 mg/L和0.12~0.38 mg/L,而48次/d组分别为0.33~0.56 mg/L和0.05~0.09 mg/L。试验结果显示,提高水循环率可降低系统中氨氮和亚硝酸盐氮的积累速度,优化养殖水质,减小水中有害物质对大菱鲆的胁迫作用,从而加快大菱鲆的生长速度,但对化学需氧量(COD)的去除没有显著影响。     

Effects of dietary organic trace mineral mixture levels on survival,growth performance,body composition and antioxidant capacity of juvenile turbot (Scophthalmus maximus)

A 70‐day feeding trial was conducted to investigate effects of dietary organic trace mineral (OTM) mixture levels on survival, growth performance, body composition and antioxidant capacity of juvenile turbot (Scophthalmus maximus). The commercial diet with 0.03% inorganic trace mineral (ITM) premix was used as the control, and other four experimental isonitrogenous (50% crude protein) and isolipidic (12% crude lipid) diets were formulated to contain 0.0375%, 0.075%, 0.1125% and 0.15% OTM mixture respectively. Results showed that there were no significant differences in survival among dietary treatments. However, the growth was not significantly different between the control and OTM supplementation diets. Especially, turbot fed the diet with 0.075% OTM had significantly higher specific growth rate than 0.15%, 0.1125% and 0.0375% OTM treatments (p < .05). Moisture, crude protein, crude lipid and ash of the whole fish body showed no significant differences among dietary treatments. Activities of superoxide dismutase in serum of turbot fed diets with 0.1125% and 0.15% OTM were significantly higher than 0.0375% OTM treatment (p < .05). Activities of total antioxidant capacity and catalase in serum were the highest, while the malondialdehyde content was the lowest when the turbot were fed the diet with 0.075% OTM. In summary, results of the present study suggested that the supplementation of 0.0375% or more OTM in diets could obtain similar performance to the commercial diet with ITM premix, and the optimal amount of OTM supplementation in diets of turbot was approximately 0.075% on basis of growth performance and antioxidant capacity.     

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.     

Effects of dietary chitosan oligosaccharide complex with rare earth on growth performance and innate immune response of turbot,Scophthalmus maximus L.

An 8‐week‐feeding trial was conducted to investigate the effect of dietary chitosan oligosaccharide complex with rare earth (COS‐REE) on growth performance and innate immune response of turbot, Scophthalmus maximus L. (Initial average weight was (12.1 ± 0.1) g) as well as disease resistance against Edwardsiella tarda. Six practical diets (approximately 53.01% protein and 12.57% lipid) were formulated to contain graded levels (0, 75, 150, 300, 600 and 1200 mg kg^?1) of COS‐REE. Results of the present study showed that, compared to the control group (0 mg kg^?1), the specific growth rate (SGR) was significantly higher in fish fed the diet with 300 mg kg^?1 COS‐REE (P < 0.05), while the feed conversion ratio (FCR) significantly decreased (P < 0.05). The phagocytic index (PI) and the activity of super oxide dismutase (SOD) of serum in fish fed the diet with 300 mg kg^?1 COS‐REE was significantly higher than fish fed the control diet (P < 0.05), but no significant differences were observed in malondialdehyde (MDA) and hepatic metallothionein (MT) concentrations. After 8 weeks, fish were challenged by intraperitoneal injection with E. tarda, and COS‐REE‐treated fish demonstrated increased protection capability. These results suggested that COS‐REE could enhance growth, innate immunity and disease resistance in turbot, and the optimum dose was approximately 300 mg kg^?1.     

Effects of hypoxia on respiratory physiology of turbot, Scophthalmus maximus

Variations in respiratory and acid-base status were studied in turbot (Scophthalmus maximus) during progressive severe hypoxia followed by recovery under normoxic conditions. The first behavioural strategy of turbot under hypoxia was an increase in amplitude and frequency of ventilation. Consequently, standard O₂ consumption remained unchanged over a broad range of O₂ tensions, until a low critical level of 30 mmHg. The hyperventilation induced a moderate blood alkalosis, compensated by a lactic acidosis. The fact that blood pH did not decrease below control values could be explained by the retention in white muscle of most of the lactate produced and by a high capacity for H⁺ excretion. During the recovery period, the marked increase in O₂ uptake corresponding to an oxygen debt repayment, was partly related to the lactate elimination. When total energy contributions of aerobic and anaerobic processes were assessed in terms of ATP, the anaerobic contribution, estimated at the deepest hypoxia level, was higher than 20% of the total energy budget and appeared to totally compensate for the decline in aerobic metabolism. Moreover, the high value of O₂ tension in arterial blood in normoxia and during recovery from hypoxia showed high diffusing capacity of gills in turbot. Our results explain the high tolerance of turbot for O₂ deficient waters.     

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.