Effects of rearing temperature and dietary short‐chain fructooligosaccharides supplementation on allochthonous gut microbiota,digestive enzymes activities and intestine health of turbot (Scophthalmus maximus L.) juveniles

The gut microbiomes of fish play important roles in host development, digestion and health. Evidence suggests that abiotic factors, such as diet and rearing temperature, could affect fish gut microbiota. In this study, the effect of dietary short‐chain fructooligosaccharides (scFOS) on turbot intestinal health, microbiota and digestive enzymes was investigated at two rearing temperatures: 15 and 20 °C. Four practical diets were supplemented with scFOS at 0, 5, 10 and 20 g kg^?1. scFOS did not affect fish performance. PCR‐DGGE did not show differences in bacterial profiles between dietary treatments; however, the number of operational taxonomic units, richness and diversity were higher at 20 °C. Enzyme activities in the foregut were not affected by rearing temperature, while in the hindgut, enzyme activities were higher at 15 °C. Total alkaline protease, α‐amylase and lipase activities in the foregut were higher in fish fed 20 g kg^?1 scFOS. Prebiotic supplementation had no effect on hindgut α‐amylase activity, while lipase activity of fish reared at 20 °C was higher in fish fed diet with 5 g kg^?1 scFOS. No differences were observed in intestinal morphology. This was the first study to simultaneously evaluate the effect of dietary prebiotic level and rearing temperature on fish intestinal microbiota and health.     

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.     

Fish protein hydrolysate affected amino acid absorption and related gene expressions of IGF‐1/AKT pathways in turbot (Scophthalmus maximus)

The effects of fish protein hydrolysate (FPH) on growth, peptide and amino acid (AA) transporters, postprandial free AA and related gene expression of IGF‐1/AKT pathway were evaluated in turbot (Scophthalmus maximus). Three diets were formulated to contain the same low level of fishmeal; meanwhile 0, 45 and 180 g/kg FPH were, respectively, supplemented to the FF (FPH‐free), FL (FPH‐Low) and FH (FPH‐High) diets. Fish fed the FH diet improved the growth compared with the other groups. For peptide and AA transporters, PepT1, B⁰AT1, CAT1 and PAT1 mRNA levels in proximal or distal intestine decreased in fish fed the FH diet. The concentration of free total essential AAs in serum was higher in the FH treatment than that in the FF treatment at 2 and 6 hr after feeding. For IGF‐1/AKT pathway in muscle, IGF‐1, 4E‐BP1 and FoxO1 mRNA levels were the highest in the FH group, whereas IGF‐1R mRNA levels were the highest expression level in the FF group. In conclusion, down‐regulated AAs transport, alleviated the delayed postprandial peak of serum‐free AAs and increased muscle protein synthesis were observed to improve the growth when turbot was fed high FPH level diets containing a high plant protein.     

Effects of essential fatty acid-deficient diets on growth, mortality, tissue histopathology and fatty acid compositions in juvenile turbot (Scophthalmus maximus)

Three diets in which the lipid component was supplied either as fish oil (FO), linseed oil (LO) or olive oil (OO) were fed to duplicate groups of juvenile turbot (Scophthalmus maximus) of initial weight 1.2 g for a period of up to 12 weeks. The latter two diets resulted in a significant reduction in specific growth rate and an increased mortality compared to the FO (control) fed fish. A liver histopathology was evident in around half of the fish fed the LO and OO diets but was absent in fish fed FO. The lesion showed indications of cellular alterations consisting of foci of densely basophilic cells but without evidence of inflammatory activity. The total lipid fatty acid composition of the carcass from fish fed LO had increased percentages of 18:2n-6 and 18:3n-3, but decreased percentages of all other polyunsaturated fatty acids (PUFA) including the physiologically important 20:4n-6, 20:5n-3 and 22:6n-3, compared to fish fed FO. Almost 2/3 of the total fatty acids in the carcass of OO-fed fish were monounsaturated while the percentages of total saturated fatty acids and all other PUFA, except 18:2n-6, were significantly reduced compared to fish fed FO. Broadly similar effects on total lipid fatty acid composition were observed in liver. In the liver glycerophospholipid classes of fish fed LO, percentages of 18:2n-6, 18:3n-3 and 20:3n-3 were significantly increased whereas all C20 and C22 PUFA, with the exception of 20:5n-3 in PI, were significantly reduced compared to fish fed FO. The liver glycerophospholipids of fish fed OO all showed significantly increased total monounsaturates, 18:2n-6, 20:2n-6, 18:2n-9 and 20:2n-9 as well as reduced percentages of 20:4n-6 and 22:6n-3, compared to fish fed FO. The brain glycerophospholipids showed broadly similar changes in response to dietary treatment although the magnitude of fatty acid alterations was less than those observed in liver. The greater mortalities in the OO-fed fish compared to the LO-fed fish suggests that incorporation of 18:3n-3 into tissue phospholipids can offset losses of long-chain PUFA more effectively than incorporation of 18:1n-9. However, levels of dietary long-chain PUFA must be optimised to allow normal growth and development. We conclude that the very low flux through the fatty acid desaturase/elongase pathways in turbot is not up-regulated by diets deficient in 20:5n-3 and 22:6n-3.     

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.     

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。     

Effect of dietary protein and lipid levels on growth and body composition of juvenile turbot (Scophthalmus maximus L) reared under optimum salinity and temperature conditions

Effect of dietary protein and lipid levels on growth and body composition of juvenile turbot was determined at optimum salinity and temperature conditions of 17 g L^?1 and 19.2 °C, respectively, by using 3 × 2 (protein levels: 550, 600 and 650 g kg^?1; lipid levels: 69 and 168 g kg^?1) factorial design with three replications of each. Fish were hand‐fed to satiety twice daily throughout the feeding trial. Weight gain and specific growth rate of fish were significantly (P < 0.05) increased with increased dietary lipid level, but not by dietary protein level. Daily feed intake was significantly (P < 0.05) affected by both dietary protein and lipid levels. Feed efficiency ratio and protein efficiency ratio were significantly (P < 0.05) affected by dietary lipid level, but not by dietary protein level. Moisture content of whole body was significantly (P < 0.05) affected by dietary lipid level, but not by dietary protein level. Crude lipid content of whole body was significantly (P < 0.05) affected by dietary lipid level, but not by dietary protein level. Significantly higher 20:5n?3, 22:6n?3 and n?3 highly unsaturated fatty acids were observed in turbot fed the low lipid diet than fish fed the high lipid diet in all protein levels, but significantly lower 18:2n?6 was observed in fish fed the former compared with the latter. In considering results of growth, specific growth rate and efficiency of feed, optimum dietary protein and lipid levels for juvenile turbot seemed to be 550 and 168 g kg^?1 of the diet, respectively, under optimum salinity and temperature conditions.     

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 different dietary arachidonic acid : docosahexaenoic acid ratios on phospholipid fatty acid compositions and prostaglandin production in juvenile turbot (Scophthalmus maximus)

Five purified diets containing AA (20:4n-6) at 0.02–0.78% dry weight and DHA (22:6n-3) at 0.93–0.17% dry weight were fed to duplicate groups of juvenile turbot (Scophthalmus maximus) of initial weight 0.87 g for a period of 11 weeks. The dietary DHA:AA ratio ranged from 62 to 0.2. Incorporation of AA into liver phospholipids increased with increasing dietary AA input. Phospholipids from fish fed diets containing 0.02, 0.06 and 0.11% of dry weight as AA generally contained less AA compared to fish fed fish oil while those fed diets containing 0.35 and 0.78% of dry weight as AA had higher AA levels in their phospholipids. The highest levels of AA were found in PI but the greatest percentage increase in AA incorporation was in PE and PC. Brain phospholipid fatty acid compositions were less altered by dietary treatment than those of liver but DHA content of PC and PE in brain was substantially lower in fish fed 0.93% pure DHA compared to those fed fish oil. This suggests that dietary DHA must exceed 1% of dry weight to satisfy the requirements of the developing neural system in juvenile turbot. In both tissues, (20:5n-3) concentration was inversely related to both dietary and tissue PI AA concentration. Similar dietary induced changes in AA, EPA and DHA concentrations occurred in the phospholipids of heart, gill and kidney. PGE₂ and 6-ketoPGF₁ were measured in homogenates of heart, brain, gill and kidney. In general, fish fed the lowest dietary AA levels had reduced levels of prostaglandins in their tissue homogenates while those fed the highest level of AA had increased prostaglandin levels, compared to fish fed fish oil. In brains, the PGE₂ concentration was only significantly increased in fish fed the highest dietary AA.Abbreviations AA arachidonic acid - DHA docosahexaenoic acid - EFA essential fatty acid - EPA eicosapentaenoic acid - HPTLC high performance thin-layer chromatography - HUFA highly unsaturated fatty acid - PC phosphatidylcholine - PE phosphatidylethanolamine - PGE prostaglandin E - PGE prostaglandin E - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid - TLC thin-layer chromatography     

Dietary lysine level affects digestive enzyme,amino acid transport and hepatic intermediary metabolism in turbot (Scophthalmus maximus)

Fish Physiology and Biochemistry - Lysine is one of the most important essential amino acids in fish, especially in the feed formulated with high levels of plant ingredients. Lysine restriction...     

Effects of salinity on the growth and lipid composition of Atlantic salmon (Salmo salar) and turbot (Scophthalmus maximus) cells in culture

The direct effects of osmotic pressure (salinity) on growth performance and lipid composition were investigated in fish cells in culture. Cell lines from a relatively stenohaline marine species, turbot (Scophthalmus maximus) (TF) and an anadromous species, Atlantic salmon (AS) were cultured in media supplemented with NaCl to produce osmotic pressures varying from 300 to 500 mOsm kg⁻¹. The growth rates of the two cell lines were affected in a similar manner by the salinity of the media with the rank order for both peak cell numbers and growth rates up to the day of peak cell number being 300 > 350 > 400 > 450 > 500 mOsm kg⁻¹. Cell death occurred in both cell lines in older cultures at all salinities with the greatest loss of viable cells in media of 300 and 350 kg⁻¹. However, there were quantitative and qualitative differences between the cell lines in their lipid metabolism in response to the salinity of the media. The lipid content expressed per cell showed a positive correlation between lipid per cell and salinity in TF cells, but this was less apparent in AS cells. The percentage of total polar lipid classes increased with increasing salinity in TF cells due mainly to graded increases in the percentages of choline phospholipids. In contrast, there were no significant differences in the proportions of polar and neutral lipid classes with salinity in AS cells. The only significant effect of salinity in AS cells was a decreased proportion of dimethylacetals in total lipid at the highest salinity. The same significant effect of salinity on dimethylacetal content of total lipid was observed in TF cells. However, in addition there was a graded decrease in the percentage of 18:2n-9 in TF cell total lipid with increasing salinity. This was accompanied by increased percentages of total n-3 and n-6 PUFA with higher proportions of both groups of PUFA at 450 and 500 compared with 300 mOsm kg⁻¹. The results show that environmental salinity, in the absence of hormonal or other physiological stimuli, has direct effects on the growth and lipid metabolism of fish cells and that these effects differ in cells from different fish species.     

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.     

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.     

Fishmeal replacement by mixed plant proteins and maggot meal on growth performance,target of rapamycin signalling and metabolism in juvenile turbot (Scophthalmus maximus L.)

Fishmeal could only be replaced by plant proteins at limited levels in aquafeeds, especially for carnivorous fish. In this study, an experiment was designed to evaluate the possibility of improving the utilization of plant proteins by maggot meal supplementation in turbot diet. Five diets were formulated: a reference diet (FM) containing 63% fishmeal and four experimental diets (35(0%), 35(3%), 40(0%), 40(3%)) in which fishmeal was substituted at different levels by plant proteins with 0–3% maggot meal. Turbot (4.90 ± 0.03 g) was fed with these diets for 9 weeks. Fishmeal was successfully replaced by plant proteins in turbot diet without growth reduction at 35% but not 40%. However, maggot supplementation (3%) at 40% plant protein replacement level achieved comparable growth performance with that of fishmeal. Maggot meal supplementation improved apparent digestibility coefficients, plasma hydroxyproline levels, intestine trypsin activities and activated target of rapamycin (TOR) signalling, all of which were decreased or down‐regulated after high plant protein replacement. Therefore, this study demonstrated maggot meal, a potential valuable protein source for turbot.     

Effect of triploidy on muscle cellularity and flesh quality of turbot (Scophthalmus maximus)

Production of triploid turbot is a relevant goal for the aquaculture industry due to the benefits associated with their sterility and growth. In order for a better understanding of the final quality of the species, muscle cellularity and quality parameters of the flesh were studied in seven diploid and seven triploid turbot after sexual maturation. White muscle cellularity was evaluated throughout white muscle cross‐sectional area from the epaxial left muscle section of each fish. To ascertain the flesh quality, several physico‐chemical parameters (moisture, ash, protein, total fat, fatty acids and minerals) were analysed, and textural mechanical properties (springiness, hardness, chewiness, cohesiveness, gumminess and adhesiveness) were determined objectively with a texturometer. Muscle cellularity was different between both groups, such that muscle fibre density was higher for diploid specimens (P < 0.001) and muscle fibres diameters were higher for triploids (P < 0.001). Both ploidy groups showed no significant differences in moisture, ash, protein and total fat content (P > 0.05). All textural properties analysed showed no significant differences between both ploidy groups (P > 0.05). In both ploidy groups, fibre density and fibre diameter showed no significant correlations (P > 0.05) with textural parameters such as springiness, cohesiveness, chewiness and adhesiveness. Ploidy did not affect fatty acid composition, with the exception of eicosapentaenoic acid (C_20:5ω3) which was higher in the triploid group. No significant differences (P > 0.05) existed between diploid and triploid turbot in content of all minerals analysed. According to our results, the ploidy status determines significant variations on muscle cellularity but does not influence the flesh quality of the turbot.     

不同培养液对大菱鲆致病性纤毛虫——贪食迈阿密虫种群增长及复壮毒力的影响

从山东烟台地区某大菱鲆养殖场的患病大菱鲆体内分离出致病性纤毛虫——贪食迈阿密虫,筛选了4种盾纤毛虫常用体外培养液[大肠杆菌培养液(DH)、牛肉浸膏培养液(BEV)、米粒培养液(RV)、改良L-15培养液(L-15)],研究了不同培养基对纤毛虫种群增长及复壮毒力的影响.试验结果显示:在BEV和L-15两种培养液中,贪食迈...     

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.     

Taurine alone or in combination with fish protein hydrolysate affects growth performance,taurine transport and metabolism in juvenile turbot (Scophthalmus maximus L.)

The study was conducted to investigate the effects of taurine (Tau) alone or in combination with fish protein hydrolysate (FPH) on growth performance, the expression of Tau transporter (TauT) and metabolic profile in juvenile turbot. FM, FPH0, FPH0+T, FPH10 and FPH10+T diets, respectively, contained 300, 150, 150, 80, and 80 g/kg fishmeal. FPH10 and FPH10+T diets contained 62 g/kg FPH. FPH0+T and FPH10+T diets were, respectively, prepared by supplementing the FPH0 and FPH10 diet formulations with 8 g/kg Tau. Specific growth rate was the highest in FM group and the lowest in FPH10 group. TauT mRNA levels in fish fed Tau supplemented diets were significantly lower than that in Tau unsupplemented diets. NMR‐based metabolomics analysis showed that Tau contents in liver of FPH0+T and FPH10+T were significantly higher than that of FM, FPH0 and FPH10. In muscle, Tau contents were significantly decreased in the FPH10+T versus FPH0 and the FPH10+T versus FPH10 comparisons. In conclusion, 62 g/kg FPH to replace fishmeal may not affect Tau synthesis, transport and metabolism. However, Tau supplemented alone or in combination with a certain level of FPH could reduce the requirement for Tau synthesis and transport and increased Tau levels in muscle and liver.