Neutral lipase and phospholipase activities in scallop juveniles (Pecten maximus) and dietary algae

Activities of the digestive enzymes neutral lipase and phospholipase A₂ were present in 2‐mm scallop juveniles, indicating an endogenous ability to digest lipids. When determined in the dietary microalgae, lipolytic activities of both enzymes differed significantly, being highest in Isochrysis galbana and Tetraselmis sp. and lowest in Chaetoceros mülleri. Compared with the activities found in scallop juveniles, enzyme activities from the microalgae were rather low and did not seem to contribute considerably to their digestive process. However, ingested algae seemed to have a regulating effect on the phospholipase activity in scallops, as juveniles fed I. galbana had a significantly lower activity than juveniles of the other treatments. No such effect was evident for neutral lipase, indicating different regulation mechanisms for both enzymes. Dietary effects on digestive enzymes in scallop juveniles should be considered when choosing new potential algal species as feed or for future formulation of artificial diets.     

On the relationship between low winter temperatures and mortality of juvenile scallops, Pecten maximus L., cultured in western Norway

The mortality of juvenile scallops (Pecten maximus L.) cultured at two densities at 8‐ and 15‐m depth was investigated at 17 trial locations along the western Norwegian coast between latitudes 58 and 65°N. During the winter of 1995/1996, scallops suffered high mortality at many sites. The results show a clear relationship between minimum temperature recorded on the sites and scallop mortality. All the scallops died at sites where minimum temperatures were lower than 2°C, while a mortality rate below 30% was only obtained at locations where the minimum temperature was 4–5°C. At temperatures between 2 and 4°C, mortality was intermediate, ranging from 52% to 100%. There were no mortality differences between culture densities, while a significantly higher mortality at 8‐ than at 15‐m depth was shown for two of the four sites with a total mean mortality lower than 30%. The results are discussed in relation to low‐temperature conditions along the Norwegian coast to suggest risk assessments for site selection in scallop culture.     

Nursery growth, survival and chemical composition of great scallop Pecten maximus (L.) spat from different larval settlement groups

Scallop Pecten maximus spat (1.3–2.1 mm shell height) from different settlement groups were transferred from hatchery to land‐based nursery at different ages and sizes. Chemical content, growth and survival were compared at transfer time and after 1 and 8 weeks of nursery growth. Growth was lowest and mortality highest in the first week after transfer. Mean shell height growth was 21.5–71.4 μm day^?1 and ash‐free dry weight (AFDW) growth ?2.7 to 10.3 μg day^?1. Spat from the first settlement group attained a larger size and weight than spat from larvae settled 3 days later, but had a lower daily growth rate (%). Keeping the late‐settled spat a longer time in the hatchery to reach a bigger size before transfer seemed not to improve subsequent nursery growth. Survival showed a large variation with mean survival ranging from 32% to 74%. A substantial reduction in lipid content was found after transfer to the nursery. Sterol content at transfer was the only lipid class correlating with survival in the nursery. Based on the results, it is justified that spat groups of different settlement age are included in production of 15‐mm great scallop spat if they are transferred from the hatchery at the same age.     

Stress and mortality in the supply chain of live scallops Pecten maximus L., from scuba diver to market

Here, we report results from a project where the aim was to study stress and mortality in the supply chain from harvest to market for live scallops Pecten maximus L., hand‐picked by scuba divers. With experiments conducted both during the warm (September) and cold (April) seasons, emphasis was put on how mechanical influence, stacking height, post‐harvest water storage and dry transport affected scallop fitness. When scallops were exposed to mild‐ or hard mechanical shocks (dropped three times from 50 or 150 cm), it was found that hard mechanical shock caused up to 90% mortality during subsequent water storage. Results from the experiments where scallops were stored at different stacking heights (15, 30 and 50 cm) in water showed that both the duration of storage and the stacking height had a significant effect on mortality. Higher stacking heights also affected survival in subsequent transport. For long‐term storage (>7 days), it is recommended to limit the stacking height of scallops stored in tanks to reduce risk of mortality. Respiration experiments showed an average oxygen consumption of 0.10 and 0.24 mg O₂ min^?1 kg^?1 f.w. in the cold and warm season respectively. Water supply during storage of scallops should be adjusted to water temperature and biomass. Registration of mortality and analysis of the Adenylic Energy Charge of scallops followed through the production cycle from harvest to market indicate that it is the sum of stressors that are determinable for final survival.     

Effects of dietary vitamin E on antioxidant defence mechanisms of juvenile turbot (Scophthalmus maximus L.), halibut (Hippoglossus hippoglossus L.) and sea bream (Sparus aurata L.)

In order to enhance growth, survival and quality during early juvenile stages of marine fish it is important to avoid lipid oxidation problems that are known to cause pathologies and disease. The aim of the present study was to characterize and compare the antioxidant systems in juvenile marine fish of commercial importance in European aquaculture, namely turbot (Scophthalmus maximus), halibut (Hippoglossus hippoglossus) and gilthead sea bream (Sparus aurata). The experiment investigated the interaction of the dietary antioxidant micronutrient, vitamin E, with antioxidant defence systems. Fish were fed with diets of identical unsaturation index supplemented with graded amounts of vitamin E. The relationships between dietary and subsequent tissue vitamin E levels were determined as well as the effects of vitamin E supplementation on lipid and fatty acid compositions of both liver and whole fish, on the activities of the liver antioxidant defence enzymes, and on the levels of liver and whole body lipid peroxidation products, malondialdehyde (thiobarbituric acid reactive substances, TBARS) and isoprostanes. Growth and survival was only significantly affected in sea bream where feeding the diet with the lowest vitamin E resulted in decreased survival and growth. A gradation was observed in tissue vitamin E and polyunsaturated fatty acid (PUFA)/vitamin E levels in response to dietary vitamin E levels in all species. The activities of the main radical scavenging enzymes in the liver, catalase, superoxide dismutase and glutathione peroxidase generally reflected dietary and tissue vitamin E levels being highest in fish fed with the lowest level of vitamin E. The indicators of lipid peroxidation gave consistent results in all three species, generally being highest in fish fed with the unsupplemented diet and generally lowest in fish fed with the diet with highest vitamin E. In this respect, isoprostane levels generally paralleled TBARS levels supporting their value as indicators of oxidative stress in fish. Overall the relationships observed were logical in that decreased dietary vitamin E led to decreased levels of tissue vitamin E, and generally higher activities of the liver antioxidant enzymes and higher levels of lipid peroxides.     

Effects of five compound attractants in high plant‐based diets on feed intake and growth performance of juvenile turbot (Scophthalmus maximus L.)

An experiment was conducted to evaluate the effects of five attractants on feed intake, growth performance and appetite‐regulating genes expression for juvenile turbot (Scophthalmus maximus L.). Fish were fed with seven diets: fish‐meal‐based diet (FM), fermented soybean meal replacing 45% fish meal (FSBM) and other diets with 1% compound attractant A (CA‐A) consisting of amino acids, compound attractant B (CA‐B) consisting of quaternary ammonium bases, compound attractant C (CA‐C) consisting of nucleotides, compound attractant G (CA‐G) consisting of the combination of CA‐A, CA‐B and CA‐C and blue mussel protein hydrolysate (BMH) on the basis of FSBM diet respectively. Daily feed intake in CA‐A group was significantly higher, but that in CA‐B group was significantly lower than FSBM group. Similarly, specific growth rate was significantly enhanced in CA‐A and CA‐G groups and were significantly decreased in CA‐B group. Protein efficiency ratio and protein retention in CA‐B group were significantly lower than FSBM group. Furthermore, BMH supplement increased the mRNA expression of Ghrelin in the intestine of fish. These results showed that CA‐A and CA‐G supplement could enhance growth in plant‐based diets, but CA‐B supplement had the opposite effect, which might be associated with lower feed intake and lower feed utilization.     

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.     

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.     

1H NMR‐based metabolomics studies on the effect of size‐fractionated fish protein hydrolysate,fish meal and plant protein in diet for juvenile turbot (Scophthalmus maximus L.)

This study was designed to evaluate changes in the metabolic profile of liver and muscle of turbot (Scophthalmus maximus L.) fed fishmeal‐based diet, diets containing size‐fractionated fish protein hydrolysate and plant protein‐based diet using ¹H NMR‐based metabolomics approach combined with the growth. Fish protein hydrolysate (FPH) was obtained by enzymatic treatment, permeate fraction was obtained as UF by ultrafiltered step, and retentate fraction was retained as RF. FM diet contained fish meal used as a single protein source. Four other diets (PP, UF, FPH and RF) contained 180 g kg^?1 diet fish meal. 54, 55 and 55 g kg^?1 dry diet UF, FPH and RF were supplemented to UF, FPH and RF diets. All diets were formulated to be isolipidic and isonitrogenous fed to five triplicate groups of turbot (16.05 ± 0.03 g) for 68 days. O‐PLS‐DA in FM versus UF, FM versus FPH, FM versus RF and FM versus PP resulted in a reliable model for muscle and liver tissue, while O‐PLS‐DA in UF versus FPH and UF versus RF only showed metabolites changes in liver tissue. Results indicated that metabolite changes among the different treatments were consistent with the growth tendency.     

Effects of diet supplementation of soya‐saponins,isoflavones and phytosterols on Atlantic salmon (Salmo salar,L) fry fed from start‐feeding

A 14‐week trial was conducted to investigate the effects of antinutritional factors (ANFs) commonly present in soybean ingredients, singly and in combination, on Atlantic salmon (Salmo salar L.) fed from start‐feeding. The experimental diets consisted of a negative control fish meal diet (FM), and a positive control diet with 167 g kg^?1 soybean meal inclusion (SBM) and four diets based on the FM diet supplemented with 2 g kg^?1 soya‐saponins (SAP), 1.5 g kg^?1 isoflavones (IFL), 0.3 g kg^?1 phytosterols (PHS) or a mixture of these (MIX). Fish fed the SAP diet showed significantly higher growth performance than those fed FM, while the IFL treatment significantly decreased growth performance of salmon fry. Fish fed the IFL diet had significantly lower maltase activity and higher trypsin activity in proximal intestine than fish fed the FM diet. Histological differences were observed in the liver of fish fed the IFL diet, characterized by reduced size of the hepatocytes. Fish fed the PHS and IFL diets showed the highest frequencies of skeletal deformities among the six treatments. In conclusion, the results indicate that purified isoflavones may negatively affect growth performance, intestinal function, liver metabolism and bone formation of salmon fry.     

The effect of ultrafiltered fish protein hydrolysate levels on the liver and muscle metabolic profile of juvenile turbot (Scophthalmus maximus L.) by 1H NMR‐based metabolomics studies

A ¹H NMR‐based metabolomics approach was to explore the effect of ultrafiltered fish protein hydrolysate (UF) levels on the liver and muscle metabolic profile of juvenile turbot (Scophthalmus maximus L.). Fish protein hydrolysate (FPH) was produced from by‐products by enzymatic treatment, and UF was obtained by diluting FPH followed filtration. Fish were fed diets containing fish meal protein, which was, respectively, replaced by UF protein 0, 50, 100, 150 and 200 g kg^?1 of dietary total protein (UF‐0, UF‐5, UF‐10, UF‐15 and UF‐20) for 68 days. OPLS‐DA of liver and muscle showed that high levels of UF in diets may lead to corresponding tissue metabolites changes. In liver tissue, changing metabolites included dimethylamine, N,N‐dimethylglycine, valine, isoleucine, leucine, arginine and alanine and were mainly involved in choline metabolism and amino acid metabolism. For muscle tissue, changing metabolites included lactate, alanine, proline, fumarate, tyrosine, histidine, cystathionine and taurine. And the metabolites were mainly involved in glycolysis and gluconeogenesis, proline metabolism and taurine metabolism. ¹H NMR‐based metabolomics is a useful approach to investigate different levels of UF on metabolic profile in liver and muscle tissues combined with the growth.     

Effects of dietary phospholipids and highly unsaturated fatty acids on the precocity,survival, growth and hepatic lipid composition of juvenile Chinese mitten crab,Eriocheir sinensis (H. Milne‐Edwards)

Precocious puberty is one of the major constraints to the further development of Chinese mitten crab (Eriocheir sinensis) farming industry. Although dietary phospholipids (PL) and highly unsaturated fatty acids (HUFA) supplementation have been shown to enhance the growth of larval E. sinensis in other studies, it is still unknown whether this also leads to a higher precocity rate for juvenile E. sinensis. This study was conducted to investigate the effects of dietary PL and HUFA on precocity, survival, growth and hepatic lipid composition of juvenile E. sinensis. Two diets were formulated with PL [3.95% dry weight (DW)] and HUFA (0.98% DW) supplementation (diet A) and without PL and HUFA supplementation (diet B) and fed to juvenile E. sinensis. Although dietary PL and HUFA levels did not significantly affect the survival and growth performance of juvenile E. sinensis, compared with crabs fed diet A, a higher precocity rate was found among juvenile E. sinensis fed diet B (P=0.051). A higher total lipid content, but significantly lower levels of HUFA and PL (P<0.05) were found in the hepatopancreas of crabs fed diet B than in those fed diet A. Meanwhile, the precocious females had significantly lower hepatosomatic index, arachidonic acid (20:4n‐6), eicosapentaenoic acid (20:5n‐3) and docosahexaenoic acid (22:6n‐3) contents in their hepatopancreas when compared with that of the normal immature juveniles (P<0.05). The results suggest that the occurrence of precocious puberty among farmed juvenile E. sinensis could be reduced by the inclusion of appropriate level of dietary PL and HUFA.     

Influence of replacing fish meal with enzymatic hydrolysates of defatted silkworm pupa (Bombyx mori L.) on growth performance,body composition and non‐specific immunity of juvenile mirror carp (Cyprinus carpio var. specularis)

This study was conducted to investigate the effects of dietary supplementation of enzymatic hydrolysates of defatted silkworm pupa (EHDSP) on growth performance, body composition and non‐specific immunity of juvenile mirror carp (Cyprinus carpio var. specularis). The control diet (EHDSP0) was produced using fish meal (FM) as the main protein source and the other four diets were formulated by substitution of 25% (EHDSP25), 50% (EHDSP50), 75% (EHDSP75) and 100% (EHDSP100) FM with EHDSP. Five groups containing 270 juvenile mirror carp (14.51 ± 0.03 g) were fed to apparent satiation for 8 weeks. The results indicated that the special growth rate, weight gain, protein efficiency ratio and feed conversion rate of experimental fish in EHDSP25 and EHDSP50 groups were not significantly different from EHDSP0 group (p > .05). The spleen index of experimental fish in EHDSP25 group was significantly higher than that of EHDSP0 group (p < .05). The muscle protein content of experimental fish in EHDSP25 and EHDSP50 groups was significantly higher than that of EHDSP0 group (p < .05). Serum alanine aminotransferase and total cholesterol of experimental fish fed with the EHDSP were significantly lower than that of control (p < .05). The activity of serum superoxide dismutase of experimental fish in EHDSP25, EHDSP50 groups was significantly higher than that of EHDSP0 group (p < .05). Intestinal trypsin activity of EHDSP25 group was significantly higher than that of EHDSP0 (p < .05). In conclusion, EHDSP can be included into diet to replace 50% FM of juvenile mirror carp without negative effect on growth, furthermore, it can improve the non‐specific immunity and function of intestinal tract.