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The global economy is increasingly focusing on the Oceans to meet its needs, for which maritime spatial planning is being fostered to promote this is done sustainably. Thus, assessing the current state and future development trends of the maritime sectors is key to evaluating the performance of the planning process. This is the aim of this study, in which the main maritime sectors related to the blue economy present in the archipelagos of the European Macaronesia Sea Basin, i.e. the Azores and Madeira (Portugal), and the Canary Islands (Spain), were identified and studied. The following maritime sectors were analysed: fisheries, marine aquaculture, marine biotechnology, coastal and maritime tourism, maritime transport, ship repair and maintenance, extraction of aggregates, deep-sea mining, offshore oil and gas, renewable ocean energies, and desalination. As part of the PLASMAR project, partners undertook in 2017–19 a literature review gathering scientific papers, official statistics and reports, which were complemented by interviews with experts from the economic sectors and public administration departments. Results show that while some maritime sectors are well established, others are emerging sectors with varying long-term growth potential. Besides, development patterns vary across the archipelagos. This study represents the first effort in the Macaronesia Sea Basin to set the basis of the current and future development conditions of the blue economy in applicability to maritime spatial planning processes at a regional level. The future post-pandemic context will provide a unique opportunity to promote the blue economy sectors and activities through the support provided by the EU Green Deal and Recovery and Resilience Plans, as well as with the actions envisioned under the United Nations Decade of Ocean Science for Sustainable Development (2021–2030). The European Macaronesia example –given its wide maritime territory, commitment, and capacities in terms of expertise and networking– presents a great potential to act as a good practice to extrapolate the new approach for a sustainable blue economy to other similar geographical settings such as island regions/states. [ABSTRACT FROM AUTHOR]

Optimising the amino acid supply in tune with the requirements and improving protein utilization for body protein growth with limited impacts on the environment in terms of nutrient loads is a generic imperative in all animal production systems. With the continued high annual growth rate reported for global aquaculture, our commitments should be to make sure that this growth is indeed reflected in provision of protein of high biological value for humans. The limited availability of fish meal has led to some concerted efforts in fish meal replacement, analysing all possible physiological or metabolic consequences. The rising costs of plant feedstuffs make it necessary to strengthen our basic knowledge on amino acid availability and utilization. Regulation of muscle protein accretion has great significance with strong practical implications. In fish, despite low muscle protein synthesis rates, the efficiency of protein deposition appears to be high. Exploratory studies on amino acid flux, inter-organ distribution and particularly of muscle protein synthesis, growth and degradation and the underlying mechanisms as affected by dietary factors are warranted. Research on specific signalling pathways involved in protein synthesis and degradation have been initiated in order to elucidate the reasons for high dietary protein/amino acid supply required and their utilization. [ABSTRACT FROM AUTHOR]

On page 24, Section 3.1.2, first paragraph, line 15, the citation [408] should be changed to [411] On page 24, Section 3.1.2, first paragraph, line 16, the citation [365] should be changed to [367]. On page 23, Section 3.1.1, second paragraph, line 11, the citation for "...Atlantic salmon, 0.6 (Lall and Bishop, 1977)" should be changed as "...Atlantic salmon, 0.6 [225]". Text Corrections On page 4, Section 2.1, first paragraph, line 8, the word "monooxygenase" should be revised to "monoxygenase". [Extracted from the article]

Abstract: Effects of the complete replacement of fish meal (FM) by soy protein concentrate (SPC) in high-energy diets (23 MJ kg−1 gross energy) on intestinal and liver histology were investigated in rainbow trout (Oncorhynchus mykiss). We developed a novel morphometric method to quantify the changes at the macro- and micromorphology level. Fish were acclimatized to the diet for 90 d and were then fed 1.0 g kg BW−1 d−1 once per day for 14 d before 10 fish per diet were randomly sampled, 6 h after their last meal. Fold height, epithelium length, stroma proportion of the proximal and distal intestines were not affected by the diet. No significant diet effect was detected on the features of the valve in the distal intestine. Fold height was linked to epithelium length in the proximal and distal intestines and this relationship was not modified by FM replacement. Enterocyte height and width were lower in SPC-fed fish in the distal intestine, but in the proximal intestine their morphology was not affected by the diet. There was no diet effect on the number of cells infiltered between the enterocytes in the proximal and distal intestines. The enterocyte nucleus was closer to the apex in the proximal and distal intestines in SPC-fed fish, but the features of the nucleus were not affected by the diet. The mean hepatocyte volume was on an average 36% lower in SPC-fed fish and was positively correlated to the hepatosomatic index for fish fed this diet only. In conclusion, SPC used in the present study did not either cause inflammatory reaction of the gut nor affected the epithelium surface. We suggest that the modifications due to SPC are linked to variations in the metabolic status of the enterocyte. [Copyright &y& Elsevier]

Our objective was to understand the reasons behind the persistent postprandial hyperglycemia in rainbow trout (Oncorhynchus mykiss). We hypothesized that in this species, high levels of dietary protein could increase the hepatic production of glucose, irrespective of the dietary carbohydrate supply. We fed juvenile rainbow trout four diets containing graded levels of protein for 14 d. Pair-feeding was employed to keep lipid and carbohydrate intakes constant. Six hours after feeding, as postulated, activities and mRNA levels of gluconeogenic enzymes (glucose-6-phosphatase, fructose-1,6-bisphosphatase) increased with increasing dietary protein (P < 0.05). However, in fish with a very low protein intake, there was a very strong increase in plasma glucose (18 mmol/L) that was also associated with a high capacity to store excess glucose as indicated by altered pyruvate kinase activity, glucokinase activity, and hepatic glycogen and fat concentrations (P < 0.05). In conclusion, at the same level of carbohydrate intake, a low dietary protein intake was associated with an unexplained increase in glycemia, which was probably responsible for the decrease in hepatic gluconeogenic enzyme expression. The effect of dietary protein on low carbohydrate utilization in this species remains unclear. [ABSTRACT FROM AUTHOR]

Simple Summary: Our aim is to introduce the mineral nutrition of fish and explain the complexity of determining requirements for these elements, which are absorbed and excreted by the fish into the surrounding water. To date, only the requirements for nine minerals have been investigated. The review is focused on the absorption and the dietary factors that reduce their absorption from feed ingredients of plant and animal origin. Some diseases, such as cataracts, anemia and bone deformity, have been linked to dietary deficiency of minerals. Aquatic animals have unique physiological mechanisms to absorb and retain minerals from their diets and water. Research and development in the area of mineral nutrition of farmed fish and crustaceans have been relatively slow and major gaps exist in the knowledge of trace element requirements, physiological functions and bioavailability from feed ingredients. Quantitative dietary requirements have been reported for three macroelements (calcium, phosphorus and magnesium) and six trace minerals (zinc, iron, copper, manganese, iodine and selenium) for selected fish species. Mineral deficiency signs in fish include reduced bone mineralization, anorexia, lens cataracts (zinc), skeletal deformities (phosphorus, magnesium, zinc), fin erosion (copper, zinc), nephrocalcinosis (magnesium deficiency, selenium toxicity), thyroid hyperplasia (iodine), muscular dystrophy (selenium) and hypochromic microcytic anemia (iron). An excessive intake of minerals from either diet or gill uptake causes toxicity and therefore a fine balance between mineral deficiency and toxicity is vital for aquatic organisms to maintain their homeostasis, either through increased absorption or excretion. Release of minerals from uneaten or undigested feed and from urinary excretion can cause eutrophication of natural waters, which requires additional consideration in feed formulation. The current knowledge in mineral nutrition of fish is briefly reviewed. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the beneficial effects of dietary arginine supplementation on growth performance, urea biosynthesis, and N excretion of juvenile gilthead sea bream ( Sparus aurata ) fed fish meal and plant-feedstuffs based diets. For that purpose, five isonitrogenous (45% crude protein) diets were formulated as follows: two diets were fish meal-based and contained 3.0 and 4.0% arginine; three diets were plant feedstuff-based and contained 1.6, 3.0, and 4.0% arginine. Triplicate groups of gilthead seabream juveniles (24 g) were fed each diet for 10 weeks. At the end of the trial, growth performance of fish fed the fish meal-based diets was higher than that of the other groups. Dietary arginine content did not affect growth performance. Ammonia excretion was not affected by diet composition, while urea excretion was directly related with arginine intake. Carbamyl phosphate synthase activity was not detected in the liver, while arginase activity increased and ornithine carbamyl-transferase decreased with dietary arginine content. Overall, an excess of dietary arginine had no beneficial effects on growth performance and feed utilisation of gilthead seabream juveniles fed plant feedstuffs-based diets. A functional urea cycle was not detected in the liver of gilthead sea bream juveniles. [ABSTRACT FROM AUTHOR]

The link between dietary carbohydrate/protein and de novo lipogenesis (DNL) remains debatable in carnivorous fish. We aimed to evaluate and compare the response of hepatic lipogenic gene expression to dietary carbohydrate intake/glucose and dietary protein intake/amino acids (AAs) during acute stimulations using both in vivo and in vitro approaches. For the in vivo trial, three different diets and a controlled-feeding method were employed to supply fixed amount of dietary protein or carbohydrate in a single meal; for the in vitro trial, primary hepatocytes were stimulated with a low or high level of glucose (3 mM or 20 mM) and a low or high level of AAs (one-fold or four-fold concentrated AAs). In vitro data showed that a high level of AAs upregulated the expression of enzymes involved in DNL [fatty acid synthase (FAS) and ATP citrate lyase (ACLY)], lipid bioconversion [elongation of very long chain fatty acids like-5 (Elovl5), Elovl2, Δ6 fatty acyl desaturase (D6D) and stearoyl-CoA desaturase-1 (SCD1)], NADPH production [glucose-6-phosphate dehydrogenase (G6PDH) and malic enzyme (ME)], and transcriptional factor sterol regulatory element binding protein 1-like, while a high level of glucose only elevated the expression of ME. Data in trout liver also showed that high dietary protein intake induced higher lipogenic gene expression (FAS, ACLY, and Elovl2) regardless of dietary carbohydrate intake, while high carbohydrate intake markedly suppressed the expression of acetyl-CoA carboxylase (ACC) and Elovl5. Overall, we conclude that, unlike rodents or humans, hepatic fatty acid biosynthetic gene expression in rainbow trout is more responsive to dietary protein intake/AAs than dietary carbohydrate intake/glucose during acute stimulations. This discrepancy probably represents one important physiological and metabolic difference between carnivores and omnivores. [ABSTRACT FROM AUTHOR]

In the present study, we evaluated the dietary fishmeal (FM) replacement potential of String Pro®, a novel methanotrophic bacterial protein meal (BM), in rainbow trout, based on growth, feed utilization, digestibility, nutrient balance and histological observations. Five isonitrogenous (49.4–51.1%) and isoenergetic (21–22.1 MJ/kg) practical diets containing 0, 12.5, 25, 37.5 and 48% BM, corresponding to 0, 25, 50, 75 and 100% FM replacement, respectively, were fed to triplicate groups of rainbow trout juveniles for 9 weeks. At the end of the feeding trial, growth rate and feed utilization were high in all the dietary groups (i.e. thermal growth coefficient of 2.55–2.96 and feed efficiency of 1.2–1.3). Feed efficiency and protein efficiency ratio did not differ between dietary groups; however, the decreasing growth trend due to reduction in feed intake was reflected in body condition and carcass estimates of protein and lipid retention, gain and whole‐body content, which decreased with increasing BM inclusion levels. There was no significant difference in apparent digestibility of protein, although, dry matter digestibility was reduced at the 48% dietary BM inclusion. Intestinal leucine aminopeptidase activity decreased; whereas, intestinal alkaline phosphatase activity and phosphorus retention increased with BM inclusion, with favourable reduction in phosphorus loss. Histological observations of the gastrointestinal tract, liver and kidney did not show any differences between dietary treatments. Based on these findings, we conclude that 12.5% of this novel BM ingredient can be safely included in the diets of rainbow trout, as a sustainable substitute for fish meal. [ABSTRACT FROM AUTHOR]

Abstract: The aim of the present work was to determine if a plant protein-based diet containing vegetable oils (VO) as the major lipid source could alter the distribution of IgM immunoreactive cells (IRCs) and the IgM expression pattern in the intestine and haematopoietic tissues of gilthead sea bream (GSB) (Sparus aurata) challenged with the myxosporean Enteromyxum leei. In a first trial (T1), GSB fed for 9 months either a fish oil (FO) diet or a blend of VO at 66% of replacement (66VO diet) was challenged by exposure to parasite-contaminated water effluent. All fish were periodically and non-lethally sampled to know their infection status. After 102 days of exposure, samples of intestine and head kidney were obtained for IgM expression and immunohistochemical detection (IHC). Additional samples of spleen were taken for IHC. Fish were categorized as control (C, not exposed), and early (E), or late (L) infected. The 66VO diet had no effect on the number of IgM-IRCs in any of the tissues or on IgM expression in C fish, whereas the infection with E. leei had a strong effect on the intestine. A combined time-diet effect was also observed, since the highest expression and IRCs values were registered in the posterior intestine (Pi) of E-66VO fish. A positive correlation was found between IgM expression and the presence of IgM-IRCs in the Pi. The effect of the time of infection was studied more in detail in a second trial (T2) in which samples of Pi were taken at 0, 24, 51, 91 and 133 days after exposure to the parasite. A significant increase of the IgM expression was detected only in parasitized fish, and very late after exposure. These results show that the duration of the exposure to the parasite is the most determinant factor for the observed intestinal IgM increased phenotype which gets magnified by the feeding of a high VO-based diet. [Copyright &y& Elsevier]

Previous studies in two rainbow trout lines divergently selected for lean (L) or fat (F) muscle suggested that they differ in their ability to metabolise glucose. In this context, we investigated whether genetic selection for high muscle fat content led to a better capacity to metabolise dietary carbohydrates. Juvenile trout from the two lines were fed diets with or without gelatinised starch (17.1%) for 10weeks, after which blood, liver, muscle and adipose tissues were sampled. Growth rate, feed efficiency and protein utilisation were lower in the F line than in the L line. In both lines, intake of carbohydrates was associated with a moderate postprandial hyperglycaemia, a protein sparing effect, an enhancement of nutrient (TOR-S6) signalling cascade and a decrease of energy-sensing enzyme (AMPK). Gene expression of hepatic glycolytic enzymes was higher in the F line fed carbohydrates compared with the L line, but concurrently transcripts for the gluconeogenic enzymes was also higher in the F line, possibly impairing glucose homeostasis. However, the F line showed a higher gene expression of hepatic enzymes involved in lipogenesis and fatty acid bioconversion, in particular with an increased dietary carbohydrate intake. Enhanced lipogenic potential coupled with higher liver glycogen content in the F line suggests better glucose storage ability than the L line. Overall, the present study demonstrates the changes in hepatic intermediary metabolism resulting from genetic selection for high muscle fat content and dietary carbohydrate intake without, however, any interaction for an improved growth or glucose utilisation in the peripheral tissues. [ABSTRACT FROM AUTHOR]

Glucose and lipid metabolism in pancreatic islet organs is poorly characterized. In the present study, using as a model the carnivorous rainbow trout, a glucose-intolerant fish, we assessed mRNA expression levels of several genes involved in glucose and lipid metabolism (including ATP-citrate lyase; carnitine palmitoyltransferase-1 isoforms, CPT; the mitochondrial isoform of the phospho enolpyrutave carboxykinase, mPEPCK and pyruvate kinase, PK) and glucosensing (glucose transporter type 2, Glut2; glucokinase, GK and the potassium channel, K) in Brockmann bodies. We evaluated the response of these parameters to changes in feeding status (food deprived vs. fed fish) as well as to changes in the amount of carbohydrate (dextrin) in the diet. A general inhibition of the glycolytic (including the glucosensing marker GK) and β-oxidation pathways was found when comparing fed versus food-deprived fish. When comparing fish feeding on either low- or high-carbohydrate diets, we found that some genes related to lipid metabolism were more controlled by the feeding status than by the carbohydrate content (fatty acid synthase, CPTs). Findings are discussed in the context of pancreatic regulation of glucose and lipid metabolism in fish, and show that while trout pancreatic metabolism can partially adapt to a high-carbohydrate diet, some of the molecular actors studied seem to be poorly regulated (K) and may contribute to the glucose intolerance observed in this species when fed high-carbohydrate diets. [ABSTRACT FROM AUTHOR]

This study was designed to assess the effects of dietary fat levels on glucose homeostasis in rainbow trout under prolonged hyperglycaemia induced by high carbohydrate intake. Trout were fed identical amounts of one of two iso-energetic diets containing either a low (LFD, 3%) or a high fat level (HFD, 20%) and similar amounts of digestible carbohydrates (26-30%) for 14 days. While a single high fat meal reduced glycaemia compared with a low fat meal, the consumption of a high fat diet for 14 days resulted in prolonged hypergylcaemia and reduced plasma glucose clearance in response to an exogenous glucose or insulin challenge. The hyperglycaemic phenotype in trout was characterised by a reduction of the activities of lipogenic and glucose phosphorylating enzymes with a concomitant stimulation of enzymes involved in glucose production in the liver and reduced glycogen levels in the white muscle. Impaired glucose tolerance (IGT) was further associated with a significant reduction of insulin receptor substrate 1 (IRS1) protein content in muscle, and with a poor response of HFD fed fish to an exogenous insulin load, suggestive of impaired insulin signalling in trout fed with a HFD. To our knowledge, this is the first study showing that a teleost can also develop a high fat-induced IGT, characterised by persistent hyperglycaemia and reduced insulin sensitivity, established symptoms of IGT and the prediabetic insulin-resistant state in mammals. Our results also provide evidence that persistent hyperglycaemia after a high carbohydrate meal stems from a metabolic interaction between dietary macronutrients rather than from high carbohydrate intake alone. [ABSTRACT FROM AUTHOR]

Water quality assessment is a key factor in the environmental management of freshwater networks, especially those including fish farms, which need cost-effective operational tools to monitor and control their waste output. In France, current legislation specifies limits in concentrations of dissolved compounds and suspended solids at fish-farm outlets. Despite the development of mass-balance modelling tools, chemical analysis of water (hydrological method) remains the most widely used method. To understand better the environmental impact of trout farms on aquatic ecosystems and to compare waste assessment methods, we monitored 20 commercial flow-through trout farms for 24 h, and we compared data obtained with the two methods (hydrological method and mass balance modelling) by linear regression. For total nitrogen and total phosphorus, the correlation between the two methods was high; thus, considering the uncertainty of both methods, this study was not able to determine which was more accurate. The high correlation between observed ammonia concentrations and predicted total nitrogen emissions provides a coefficient for estimating ammonia emissions at the farm level. The same approach is proposed for the evaluation of phosphate emissions. In conclusion, this study confirms the utility of simulation modelling for assessing nutrient release from fish farms. [ABSTRACT FROM PUBLISHER]

The objective of the present work is to investigate the main metabolic pathways by which dispensable amino acids (DAA) are diverted towards lipid formation in blackspot seabream. For that purpose, a control diet was formulated to contain 45% of crude protein (7.2 g N/100 g dry matter) mainly supplied by fish meal (45P). In two other diets, 22.2% of the dietary nitrogen (1.6 g N/100 g dry matter) was replaced by an equivalent amount of nitrogen provided by two different mixtures of DAA: alanine and serine (diet AS) or aspartic and glutamic acid (diet AG). A fourth diet (diet 35P) only containing 35% of crude protein (5.6 g N/100 g dry matter) was included in order to analyze the possible additive effects of DAA. Compared to fish fed diet 35P, blackspot seabream appear to make a more efficient use of the nitrogen provided by alanine and serine than that provided by aspartic and glutamic acids in terms of growth. Contrary to fish fed AG, fish fed AS attained similar specific FAS activities as 45P fed fish, suggesting a further role of alanine and serine on this lipogenic pathway. Dietary nitrogen reduction (45P vs. 35P) or its replacement by a mixture of aspartic and glutamic acids (diet AG) were shown to up-regulate phosphoenolpyruvate carboxykinase (PEPCK) but without, however, any effect on plasma glucose levels. Dietary nitrogen level and nature seems to exert a complex regulation on energetic pathways through the gluconeogenesis/tricarboxylic acids cycle interaction. [ABSTRACT FROM AUTHOR]

Abstract: In order to investigate the impact of dietary lipid sources on mechanisms involved in lipid deposition, three groups of European seabass fingerlings with average initial body weight of 5.2±1.0 g were fed three diets differing only by lipid source. These diets were: 100% anchovy oil (diet A), 40% anchovy oil–60% mix of vegetable oils (35% linseed, 15% palm, 10% rapeseed) (diet B) and 40% anchovy oil–60% mix of vegetable oils (24% linseed, 12% palm, 24% rapeseed) (diet C). After 64 weeks of rearing, when seabass reached the size of 160 g, the activity of lipogenic enzymes (fatty acid synthetase, glucose-6-phosphate dehydrogenase and malic enzyme) in liver and of lipoprotein lipase (LPL) in perivisceral adipose tissue, liver and white muscle were measured. Transport of lipid by lipoproteins was examined by determining plasma lipid composition and lipoprotein classes. Dietary oil source did not modify growth performance or lipid content of flesh and liver of seabass. Replacement of 60% of fish oil by the two mixtures of vegetable oils had no significant effect on hepatic lipogenesis and activity of LPL in liver and adipose tissue. Activity of LPL in white muscle was decreased in fish fed diet C compared to those fed diets A and B. Diets containing the mixture of vegetable oils led to lowered plasma, VLDL and LDL cholesterol levels compared to diet A. It is concluded that replacing 60% of fish oil by the two mixtures of vegetable oils in the feeds of European seabass fingerlings until they reach the size of 160 g has no marked effect on growth performance, lipogenesis and tissue lipid uptake but has a hypocholesterolemic effect. [Copyright &y& Elsevier]

Understanding the environmental burdens associated with aquafeeds is a critical component for assessing and improving the environmental performance of aquaculture. The aim of the study was to assess the environmental impacts associated with feeds for rainbow trout production in France, using Life Cycle Assessment (LCA). The stages assessed are: the extraction of the raw materials, the production and transformation of the primary ingredients used, the manufacturing of the feeds, the use of the feeds at the farm, transport at all stages, and the production and use of energy resources. The assessment revealed that the use of fishery resources (such as biotic resource use) and nutrient emissions at the farm (such as eutrophication potential) contribute most to the potential environmental impacts of salmonid aquafeeds. Improvements in feed composition and management practices seem to be the best ways for improving the environmental profile of aquafeeds. [ABSTRACT FROM AUTHOR]

We examined the influence of the oligosaccharide component of a lupin (Lupinus angustifolius) meal when fed to rainbow trout. Two strategies for examining the influence of oligosaccharides were undertaken, including the use of an exogenous enzyme supplement of α-galactosidase to degrade the oligosaccharides and ethanol extraction processing to remove the oligosaccharides. Controls for both processing and degradation of the galactosides were also included in the study. The test treatments were fed to the fish as 30% of their diet and were compared against a reference diet using standard diet-dilution digestibility techniques. The removal of the ethanol-soluble component of the lupin meal had the greatest influence on its apparent digestibility, with significant effects clearly seen on the digestibility of the nitrogen, organic matter and nitrogen-free extractive (NFE) components. Addition of exogenous α-galactosidase to the lupin meal produced a significant difference in the digestibility of nitrogen and numerical improvements in the values of most nutrient parameters. Comparison of the digestibilities of the enzyme-supplemented lupin meal, the galactose-supplemented extracted lupin meal and the original lupin meal treatments indicated that rainbow trout can absorb the monosaccharides produced from the degradation of α-galactosides. This improvement was further validated by examining the influence of the ethanol extraction process, which showed that it is the removal of the ethanol-soluble components, most likely oligosaccharides, that is the contributing factor. These findings indicate that further processing of lupin meals to remove their oligosaccharide fraction would have beneficial nutritional implications for fish. [Copyright &y& Elsevier]

There have been some arguments concerning supplementary feed (cereals) based common carp production in fishponds and water pollution, mostly in Central Europe. Using Czech Republic (top producer in EU) as a benchmark and combining data on nutrient digestibility of feedstuffs used combined with analyses of literature data, we have assessed – nutrient footprint (∼9.4–10.8 kg N ha−1, ∼2.7–3.2 kg P ha−1; 1.5–4 × < EU crop-livestock sectors); nutrient utilization efficiencies (NUE N ∼36%, NUE P ∼50%; 1.5–1.7 × > EU livestock average); autochthonous nutrient removal (∼8–9.2 kg N ha−1, 1.4–1.6 kg P ha−1); eco-cost burden (13–29 × ≪ positive services); eco-services (∼74.5–100.6 million € country−1; ∼2375 € ha−1) of carp production in Central Eastern European Region (CEER). Digestible nutrients offered by natural prey (7.9% N, 1% P on dry matter basis) to carp are ∼5–8 times higher than those provided by cereals and remains the key determinant for production. Despite this, 70–90% of nutrient footprint from feeding is contributed by cereals. Neutral footprint (∼374 kg ha−1) and exclusively natural (up to 300 kg ha−1) carp production intensities were identified, following which, commercial interest of carp farming may falter (costing intangible losses >56.5 million € in CEER), despite achieving 'greener-goals'. Per production cycle, carp aquaculture in CEER fishponds offer at least 579 million € worth of services. Our results show that carp production in ponds have lesser nutrient burden than crop and livestock productions in EU. Existing management of fishponds 'barely meet' optimum P requirements of common carp and present production intensity should not be vilified as a pollution causing activity. Risks and solutions for achieving both environmental (minimized footprint) and aquaculture goals (uncompromised production) are discussed. • Arguments concerning carp production & water pollution in Central Europe. • Political lobbying between environmentalists & fishpond managers for legislations. • Carp production in fishponds 'cleaner' than other food production sectors in EU. • Footprint < EU crop-livestock; NUE > EU livestock; Eco burden ≪ positive services. • Downscaled production for greener goals ≈ intangible losses >56.5 mill € in CEER. [ABSTRACT FROM AUTHOR]

Selenium (Se) deficiency is a problem widely encountered in humans and terrestrial livestock production with increasing attention also in aquaculture. Se supports the antioxidant system, which becomes especially important during stressful conditions. In the present study, the effect of Se-supplementation in broodstock and fry diets on the performance and antioxidant metabolism of rainbow trout fry under acute hypoxia was investigated. Rainbow trout broodstock were fed plant-ingredient based diets either without any Se-supplementation (Se level: 0.3 mg/kg) or supplemented with Se supplied as sodium selenite or as hydroxy-selenomethionine (Se level: 0.6 mg/kg respectively) for 6 months prior to spawning. The progenies were subdivided into three triplicate feeding groups and fed diets with similar Se levels compared to the parental diets, resulting in a 3x3 factorial design. After 11 weeks of feeding, the fry were either sampled or subjected to a hypoxic stress challenge. One hundred fish were transferred to tanks containing water with a low oxygen level (1.7 ± 0.2 ppm) and monitored closely for 30 min. When a fish started to faint it was recorded and transferred back to normoxic water. Direct fry feeding of the hydroxy-selenomethionine supplemented diet improved the resistance towards the hypoxic stress. On the contrary, fry originating from parents fed Se-supplemented diets showed a lower stress resistance compared to fry originating from parents fed the control diet. Fry subjected to hypoxia showed elevated oxidative stress with reduced glutathione (GSH) levels and increased isoprostanes (IsoP) and phytoprostanes (PhytoP) levels produced by lipid peroxidation of polyunsaturated fatty acids (PUFA), arachidonic and α-linolenic acids respectively. Increased mRNA expression of transcription factors (nrf2 , nfκb , keap1X2) and decreased mRNA expression of antioxidant enzymes (trxr , sod , gstπ) indicated a transcriptional regulation of the antioxidant response. In stressed fry, the mRNA expression of several antioxidant genes including gr , msr and gstπ was found to be higher when fed the control diet compared to the sodium selenite treatment, with a contrary effect for parental and direct Se nutrition on gpx. The long-term parental effect becomes of greater importance in stressed fry, where more than half of the genes were significantly higher expressed in the control compared to the selenite supplemented group. Image 1 • Programming effects by parental selenium nutrition were detectable in long-term. • Parental and direct inorganic selenium feeding had opposite effects on GPX. • Hypoxic stress induced an antioxidant response in rainbow trout fry. • Direct selenium feeding improved the stress resistance towards hypoxia. • Parental selenium feeding decreased the stress resistance towards hypoxia. [ABSTRACT FROM AUTHOR]

The present study aimed to determine the effects of dietary inclusion of selenium (Se) in the form of either inorganic Se (sodium selenite, NaSe) or organic Se (hydroxy-selenomethionine, OH-SeMet) on gilthead seabream (Sparus aurata). Triplicate groups of 150 fish each (6.2 ± 0.04 g initial body weight) were distributed in 15 tanks and fed one of five experimental diets for 63 days. A control diet (C) without Se supplementation (0.8 mg Se/kg diet), was formulated and served as a basal diet for the other 4 experimental feeds, supplemented with either 0.2 or 0.5 mg Se/kg supplied in the form of NaSe or OH-SeMet. Fish growth performance, biochemical composition of liver and muscle and Se content, malondialdehyde (MDA) concentration in muscle and hepatopancreas, hepatocyte morphology and lysozyme activity in the serum were studied. Furthermore, expression of related hepatic genes, such as manganese superoxide dismutase (Mn- sod), catalase (cat), heat shock protein 70 (hsp70), and glutathione peroxidase (gpx) was also analyzed. After the trial, fish were exposed to an acute and chronic confinement stress. Blood samples for cortisol analysis were taken at 0 and 2 h after the acute stress and after 7 days of chronic stress. The highest growth rate was observed in fish fed OH-SeMet at a level of 0.2 mg/kg, but with no significant difference with fish fed the control diet with no-added Se. The lowest growth was observed in fish fed NaSe up to 0.5 mg/kg. Increase in dietary Se, particularly in the form of OH-SeMet, led to an increase in Se contents in liver and muscle. Furthermore, dietary inclusion of OH-SeMet, led to a significant (p <.05) reduction in MDA in both liver and muscle. Se inclusion as NaSe at 0.2 mg/kg was not as effective as organic Se to improve muscle oxidative status. Dietary inclusion of Se at 0.2 mg/kg significantly reduced plasma cortisol after 2 h of acute stress, regardless of the Se form fed. Lysozyme activity measured in the serum was decreased with the increase in dietary Se supplementation. In summary, supplementation of Se up to 0.2 mg/kg (1–1.1 mg/kg analyzed dietary Se), particularly in the form of OH-SeMet, led to a beneficial effect on growth, maintenance of hepatic morphology and improved protection of juvenile gilthead seabream from acute or chronic stress. Besides, OH-SeMet was found to be more effective than NaSe in protection against oxidative stress in fish muscle. • This study aims to determine the importance of dietary Se supplementation on health and welfare of gilthead seabream. • The organic Se promotes growth and protect fish against the oxidative stress in muscle and liver. • The organic Se is more efficient than the inorganic form. • Supplementation with OH-SeMet (total Se content: 1.1 mg/kg) is sufficient to promote fish growth and protect against oxidative stress. [ABSTRACT FROM AUTHOR]

We studied the effects of supplementing a plant-ingredient based rainbow trout broodstock diet with different forms of selenium (Se) on reproductive performance, parental Se transfer and antioxidant metabolism in their progeny before first-feeding. Three groups of rainbow trout were fed a diet either without selenium supplementation (NC, basal Se level: 0.3 ppm) or supplemented with 0.3 ppm Se either in the form of sodium selenite (SS) or as hydroxy-selenomethionine (SO, dietary Se level: 0.7 ppm) over a 6-month period prior to spawning. In Se-supplemented groups, the total number of females spawning was significantly higher compared to the negative control group and the females fed SO began to spawn earlier compared to females fed SS or NC. Total Se concentrations were significantly higher in female muscle of SO group. Higher Se concentrations in the oocytes of both Se-supplemented groups confirmed a maternal transfer of Se, while total Se concentrations in milt samples were not significantly different between dietary groups. There was no effect on glutathione peroxidase (GPX) or other antioxidant enzyme activity in the liver of female or male broodstock, whereas in the swim-up fry, GPX activity was significantly higher in both Se-supplemented treatments with the highest activity in the SO group. Se-supplementation enhanced the expression of hepatic SelPa in broodstock males and females along with MsrB1, GPx1a, GPx4a2, CAT, Gclc, Keap1 and MsrB2 in male liver. In swim-up fry, only organic Se-supplementation led to a higher gene expression for SelPa, GPX1a, GPX1b2, CAT and MsrB2. An improvement of the oxidative status in whole-body of swim-up fry could not be confirmed in this study as the GSH/GSSG was even lower in swim-up fry of Se-supplemented groups, whereas the 8-isoprostane levels were lower in the SS group and no effect on protein carbonyl levels could be detected. Organic Se-supplementation led to a significant increase in the α-tocopherol and vitamin C levels in the progeny. These results show that dietary Se in broodstock nutrition influences spawning occurrence and along with the parental transfer, several traits in the progeny are affected. • Dietary Se affected the number and timing of spawning in rainbow trout females. • Better bioavailability of Se in broodstock diets increased its transfer to the progeny. • Highest GPX activity and mRNA expression of selenoproteins in offspring of broodstock fed organic Se. • Organic Se supplementation increased vitamin E and C levels in progeny. [ABSTRACT FROM AUTHOR]

The aim of the study was to analyse the effects of different dietary methionine (Met) concentrations on the mechanisms of fish muscle growth (hyperplasia and hypertrophy) by using cellular and molecular approaches. Three plant-based diets differing by their Met level (MD, Met-deficient; MC, Met-control, corresponding to established Met requirement; MH, Met-high) were fed to rainbow trout juveniles for 12 weeks. Met deficiency led to a reduced somatic growth right from the first 3 weeks of feeding onwards, while feeding the MH-diet led to an increased body growth in the 6 last weeks of feeding. The protein gain of the juveniles increased with dietary Met content. The protein retention efficiency of the MD-fed juveniles was half that of the MC- and MH-fed juveniles which were statistically similar. The changes induced by MD- and MH-feeding at muscle molecular and cellular levels were different. Compared to control, dietary Met deficiency altered the expression of myogenic, muscle growth- and metabolism-related genes by increasing Myf5, MyoD1, Mrf4, Mef2a, βact, CathD, GSTπ and TNFα expressions and decreasing Col1α1 and IGF1 expressions, whereas a high level of dietary Met decreased Pax7a2, Mef2c, βact, CathD, GDH2 and NF-κB expressions. The Met-control fed fish were discrimated according to HGF, Pax7a1 and Pax7a2 expressions. Dietary Met level thus acted on the expression of genes regulating specific transition points of myogenesis, and affected the expression of muscle structural genes and growth factors involved in satellite cell activation and muscle growth. Molecular results suggested that MD-diet favours satellite cell return to quiescence, while MC-diet favours satellite cell activation and MH-diet myogenic differentiation. At cellular level, the muscle total cross-sectional area of the juveniles fed the MD-diet was 28% lower than that of the juveniles fed the MC-diet despite a similar total number of white muscle fibres, and the number of white muscle fibres with a large diameter was lower in MD-fed fish than in MC-fed ones. The muscle total cross-sectional area of the juveniles fed the MH-diet was 32% higher than that of the juveniles fed the MC-diet, their total number of white muscle fibres 25% higher, and their number of white muscle fibres with a large diameter similar. Increasing dietary methionine above established requirement thus promotes white muscle hyperplasia and muscle growth in rainbow trout juveniles, while methionine deficiency dysregulates the expression of myogenic and muscle growth-related genes and impairs white fibre hypertrophy. Statement of relevance Determining the effects of dietary methionine levels on muscle growth mechanisms will help develop low fish meal feeds. [ABSTRACT FROM AUTHOR]

Meagre (Argyrosomus regius) is a fast-growing marine teleost and a promising candidate for the diversification of the Mediterranean aquaculture industry. Information on specific dietary requirements is necessary to improve the competitiveness of meagre farming. While the essential fatty acids (EFA) requirement of meagre fingerlings was previously estimated, knowledge regarding the EFA requirement of on-growing meagre juveniles is still lacking. To assess the EFA requirement of juvenile meagre, six isolipidic, isoproteic, semi-purified diets with different levels of n-3 long-chain polyunsaturated fatty acid (n-3 LC-PUFA) contents 0.0%, 0.3%, 0.6%, 0.9%, 1.2%, and 1.5% of diet dry matter (DM) were formulated and fed three times daily to triplicate groups of juvenile meagres of 35.6 ± 0.3 g for twelve weeks. Growth rates, whole body composition and fatty acid profile, liver lipid content as well as fatty acid profiles were assessed. In addition, the incidence of hepatic steatosis and granulomatosis was evaluated. Juvenile meagre fed the 0.0% n-3 LC-PUFA diet showed significantly lower growth and final body weight compared to all other treatments. There was an increase in growth of meagre with increasing dietary n-3 LC-PUFA levels and weight gain plateaued at levels above 0.9% n-3 LC-PUFA. Occurrence of granulomas examined in the liver supported the data on growth as the hepatic granulomatosis incidence in the groups fed <0.9% n-3 LC-PUFA were greater compared to fish fed diets containing higher levels of n-3 LC-PUFA. The findings of this study showed that the dietary n-3 LC-PUFA requirement of juvenile meagre for optimal growth and liver health status lays between 0.7 and 0.8% of the dietary dry matter. • Juvenile meagre showed excellent growth rates and food conversion efficiencies. • Growth performance declined in fish fed dietary EFA levels of 0.6% DM or less. • Diets with low EFA levels caused a higher incidence of hepatic granulomas. • The dietary requirement for EFA of juvenile meagre is between 0.7 and 0.8% DM. [ABSTRACT FROM AUTHOR]

We studied the effect of changes in dietary methionine (Met) supply in broodstock and first-feeding rainbow trout fry (offspring). Three plant-based diets differing in Met level (deficient, adequate or in excess of the established requirement) were fed to the broodstock (male and female) for 6 months prior to spawning (diets BD, BA and BE, respectively). The offspring from the parental Met-groups was then challenged in turn with the different Met fry-diets (FD, FA and FE, respectively) for 3 weeks from first-feeding. At spawning, females fed diet BD had significantly higher plasma total and LDL-cholesterol and slightly lower plasma triacylglycerol. Diet BD reduced female (but not male) growth, weight of spawn and egg size, but had no effect on relative fecundity. The free amino acid profile of oocytes was modified, with levels of Met and Cys correlating positively with the Met-levels of broodstock diets. SAM and SAH levels in oocytes followed the same pattern, as opposed to SAM/SAH ratio. At the swim-up stage, no significant effect of parental diet on fry weight was noted, whereas survival was the highest in fry from BE-broodstock. The subsequent 21-day fry feeding with different Met levels highly affected the daily growth index with a significant interaction between the parental-diet and fry-diet effects. The expression of a number of genes regulating sulfur amino acid metabolism was modified either directly by the dietary Met supply in both broodstock liver and in whole fry (e.g. BHMT1, GR, GSTπ, MsrA1) or indirectly by the parental Met intakes as seen in the swim-up fry (e.g. BHMT1, MTR, GSTπ, MsrA1). Importantly, long-lasting parental effects linked to broodstock Met-intake were seen in the fry, 21-days after first-feeding and irrespective of the fry diet (CTH, MsrA1, MsrB2, SOD2). Similarly, parental effects were noted on the gene expression of both NPY and POMC feeding peptides in fry prior to exogenous feeding which persisted for POMC in the 21-day fry. Parental effects were also demonstrated on the key myogenic gene Myog, on fMHC and GDH in swim-up fry, which persisted for GDH in 21-day fry. In summary, our results demonstrate that dietary Met levels of rainbow trout broodstock affect various traits in the offspring, some of which persisted during the first weeks of exogenous feeding. Further studies need to evaluate the long-term persistence of the parental effects over time and to elucidate the mechanisms, whether epigenetic or not. Statement of relevance Determining the multiple effects of dietary methionine levels on reproductive, growth performance and metabolism in offspring will help improve formulations of low fish meal feeds for rainbow trout at sensitive life cycle stages. [ABSTRACT FROM AUTHOR]

Growth and digestibility trials were undertaken to evaluate a single cell protein (SCP) produced from methane-oxidising bacteria Methylococcus capsulatus (String Bio, Bengaluru, India) as a replacement for fishmeal in the diets of Pacific white shrimp, Penaeus vannamei. Five isonitrogenous and isoenergetic diets were formulated to contain different levels of SCP: Control (Fishmeal, no SCP), SCP inclusion at 50 g/kg (S-5), 100 g/kg (S-10), 200 g/kg (S-20), 250 g/kg (S-25). The final body weight of the shrimp increased with increasing levels of SCP up to 200 g/kg (S-20) and recorded the best feed efficiency when compared to other treatments. Nitrogen (N) and phosphous (P) gain and retentions were improved in shrimp fed diets containing upto 200 g/kg SCP. There was no significant difference in the whole-body chemical and amino acid composition of shrimp. Apparent protein digestibility of the SCP was greater than 0.90. The expression levels of immune-relevant genes (lysozyme, Toll-like receptor and immune deficiency (IMD)) were up-regulated in the shrimp fed diet S-20. Results of this work demonstrate that SCP from M. capsulatus can be efficiently used in the diet of P. vannamei up to 200 g/kg (two-thirds of fishmeal replacement) with significant beneficial effects on growth, feed and nutrient utilisation. • SCP derived from Methylococcus capsulatus is a protein rich ingredient with a comparable amino acid profile with fishmeal • The digestibility of SCP by Pacific white shrimp was very high • SCP can be used in the diets of Pacific white shrimp upto 200 g/kg without affecting growth and nutrient utilization • The innate immune response was higher in Pacific white shrimp fed diets with SCP than those fed the control diet [ABSTRACT FROM AUTHOR]

Background: The achievement of sustainable feeding practices in aquaculture by reducing the reliance on wild-captured fish, via replacement of fish-based feed with plant-based feed, is impeded by the poor growth response seen in fish fed high levels of plant ingredients. Our recent strategy to nutritionally program rainbow trout by early short-term exposure to a plant-based (V) diet versus a control fish-based (M) diet at the first-feeding fry stage when the trout fry start to consume exogenous feed, resulted in remarkable improvements in feed intake, growth and feed utilization when the same fish were challenged with the diet V (V-challenge) at the juvenile stage, several months following initial exposure. We employed microarray expression analysis at the first-feeding and juvenile stages to deduce the mechanisms associated with the nutritional programming of plant-based feed acceptance in trout. Results: Transcriptomic analysis was performed on rainbow trout whole fry after 3 weeks exposure to either diet V or diet M at the first feeding stage (3-week), and in the whole brain and liver of juvenile trout after a 25 day V-challenge, using a rainbow trout custom oligonucleotide microarray. Overall, 1787 (3-week + Brain) and 924 (3-week + Liver) mRNA probes were affected by the early-feeding exposure. Gene ontology and pathway analysis of the corresponding genes revealed that nutritional programming affects pathways of sensory perception, synaptic transmission, cognitive processes and neuroendocrine peptides in the brain; whereas in the liver, pathways mediating intermediary metabolism, xenobiotic metabolism, proteolysis, and cytoskeletal regulation of cell cycle are affected. These results suggest that the nutritionally programmed enhanced acceptance of a plant-based feed in rainbow trout is driven by probable acquisition of flavour and feed preferences, and reduced sensitivity to changes in hepatic metabolic and stress pathways. Conclusions: This study outlines the molecular mechanisms in trout brain and liver that accompany the nutritional programming of plant-based diet acceptance in trout, reinforces the notion of the first-feeding stage in oviparous fish as a critical window for nutritional programming, and provides support for utilizing this strategy to achieve improvements in sustainability of feeding practices in aquaculture. [ABSTRACT FROM AUTHOR]

Responses in micro-mineral metabolism to changes in dietary ingredient composition and inclusion of a micro-mineral premix (Fe, Cu, Mn, Zn and Se) were studied in rainbow trout. In a 2 x 2 factorial design, triplicate groups of rainbow trout (initial weight: 20g) were fed over 12 weeks at 17°C a fishmeal-based diet (M) or a plant-ingredient based diet (V), with or without inclusion of a mineral premix. Trout fed the V vs. M diet had lower feed intake, growth, hepato-somatic index, apparent availability coefficient (AAC) of Fe, Cu, Mn and Zn and also lower whole body Se and Zn concentration, whereas whole body Fe and Cu and plasma Fe concentrations were higher. Feeding the V diet increased intestinal ferric reductase activity; at transcriptional level, hepatic hepcidin expression was down-regulated and ferroportin 1 was up-regulated. Transcription of intestinal Cu-transporting ATPases and hepatic copper transporter1 were higher in V0 compared to other groups. Among the hepatic metalo-enzyme activities assayed, only Se-dependent glutathione peroxidase was affected, being lower in V fed fish. Premix inclusion reduced the AAC of Fe, Cu and Zn; increased the whole body concentration of all micro- minerals; up-regulated hepatic hepcidin and down-regulated intestinal ferroportin 1 transcription; and reduced the transcription of Cu-transporting ATPases in the intestine. Overall, the regulation of micro-mineral metabolism in rainbow trout, especially Fe and Cu, was affected both by a change in ingredient composition and micro-mineral premix inclusion. [ABSTRACT FROM AUTHOR]

Genetic selection programmes in gilthead seabream mainly focus on traits related to growth, disease resistance, skeletal anomalies, or fillet quality. However, the effect of selection for growth on the reproductive performance of seabream broodstock has not received much attention. The present study aimed to determine the effect of selection for growth traits, high (HG) or low (LG) growth, and broodstock feeding with fish oil (FO diet) or rapeseed oil (RO diet) as main lipid sources, on reproductive performance of gilthead seabream. For the first part of the spawning season (Phase I) HG and LG broodstock were fed a commercial diet and the HG broodstock produced a higher number of larvae and higher viable eggs, hatching and larval survival rates than LG broodstock, affecting egg fatty acid profiles. For the second part of the study (Phase II) broodstock were fed one of the two diets containing FO or RO. Fecundity in terms of viable eggs, hatchlings, and larvae produced, as well as fertilization rates, were improved in HG broodstock. Some fatty acids such as 18:0, 20:2n-6, 20:3n-3 or EPA/ARA were also affected by the growth selection. According to the two-way ANOVA analysis, feeding the RO diet did not significantly affect fecundity parameters, but slightly reduced fertilization and hatching rates in HG broodstock. Nevertheless, HG broodstock showed better spawning quality parameters than LG broodstock, even when they were fed the RO diet. Egg fatty acid profiles reflected diet composition, although DHA contents were not affected. In conclusion, broodstock selected for high growth had a positive effect on broodstock performance, and FO replacement by RO did not markedly affect reproduction providing that fatty acid contents were sufficient to fulfill the essential fatty acid requirements of gilthead seabream broodstock. • High growth (HG) trait broodfish exhibited improved reproductive performance. • RO diet fed to HG broodfish did not negatively affect egg and larval quality. • The egg DHA content was not affected in HG broodfish fed with RO diet. [ABSTRACT FROM AUTHOR]

Abstract: High concentrations of n−3 polyunsaturated fatty acids (PUFA) that are readily susceptible to lipid peroxidative damage are found in fish feeds and in the tissues of fish, especially in early developmental stages. A dietary phospholipid (PL) supply has been shown to be beneficial during these critical stages. The objective of the study was to characterize the response of the antioxidant defense system under dietary prooxidant conditions in presence or absence of dietary PL during early development of rainbow trout. Rainbow trout (Oncorhynchus mykiss) at the first-feeding fry stage (mean weight: 66±2mg) or at the fingerling stage (mean weight: 1.5±0.4g) were fed 4 semi-purified diets supplemented with 12% fresh fish oil or 12% oxidized fish oil and 6% soybean lecithin or 6% soybean oil for 4weeks at 17°C. At fry stages, rainbow trout fry fed the PL-supplemented diets had a significantly higher final body weight than fry fed the PL-free diets (0.37±0.07 vs. 0.27±0.03g, respectively). Dietary inclusion of oxidized lipid reduced growth (0.19±0.02 vs. 0.45±0.07 and 2.5±0.6 vs. 4.8±0.6g, respectively) and increased the mRNA expression of antioxidant enzymes such as glutathione reductase and glutathione S-transferase at both developmental stages. However, dietary control of antioxidant enzyme activities and vitamins was low in rainbow trout fry whereas increased activities of antioxidant enzymes and decreased tocopherol contents were noticed in rainbow trout fingerlings fed oxidized lipid compared to rainbow trout fry fed fresh oil. This resulted in higher content of lipid peroxidation products in rainbow trout fry fed oxidized lipid compared to fish fed fresh lipid whereas this difference was reduced at the fingerling stage. The present study demonstrates that rainbow trout fry are more susceptible to oxidative stress induced by dietary oxidized lipid than rainbow trout fingerlings, possibly due to delayed response or lack of complete development of endogenous antioxidant defense system. [Copyright &y& Elsevier]

Abstract: The aim of this work was to investigate the catabolic process of three kinds of dietary carbohydrates in the gut of sea bass juveniles, with the possible contribution of the intestinal microbiota to the nutrition of the host, and the subsequent effects on intermediary metabolism. A first diet contained waxy maize (99% amylopectin), a highly digestible form of starch. A second diet was less quickly digestible due to its high amylose content of resistant starch. Two other diets contained fibre instead of starch, either only cellulose as control, or also other non-starch polysaccharides brought by lupin meal. The effect of the diets on the host confirmed previous results, with the stimulation of glucose storage in the liver in sea bass fed the starchy diets, which caused a significant increase in liver weight, while lupin meal caused an increase in visceral mass. Glycaemia was higher 7±1h after the last meal in the group fed resistant starch, compared to the other dietary groups, while the fast digestion of waxy maize resulted already in hypertriglyceridemia, possibly due to hepatic neolipogenesis. At the same sampling time, the activity of free amylase was reduced in the intestine of sea bass fed resistant starch, but maltase activity was stimulated in the brush border membranes of enterocytes in the same group, confirming thus the timely digestion of resistant starch. Hepatic mRNA transcripts indicated that glucose metabolism was oriented towards neoglucogenesis by the high-fibre diets, and towards glucose storage by the starchy diets, especially with waxy maize. The diet influenced both faecal and mucosal microbiota, though in different ways, likely due to the interaction with the host. Lupin meal seemed potentially interesting as a source of prebiotic polysaccharides, by modifying the balance between Vibrio spp. and Clostridium sp. Both forms of starch were also partly metabolised by microbiota, resulting in an increased concentration of acetate in the faeces. [Copyright &y& Elsevier]

Sustainable aquaculture, which entails proportional replacement of fish-based feed sources by plant-based ingredients, is impeded by the poor growth response frequently seen in fish fed high levels of plant ingredients. This study explores the potential to improve, by means of early nutritional exposure, the growth of fish fed plant-based feed. Rainbow trout swim-up fry were fed for 3 weeks either a plant-based diet (diet V, V-fish) or a diet containing fishmeal and fish oil as protein and fat source (diet M, M-fish). After this 3-wk nutritional history period, all V- or M-fish received diet M for a 7-month intermediate growth phase. Both groups were then challenged by feeding diet V for 25 days during which voluntary feed intake, growth, and nutrient utilisation were monitored (V-challenge). Three isogenic rainbow trout lines were used for evaluating possible family effects. The results of the V-challenge showed a 42% higher growth rate (P = 0.002) and 30% higher feed intake (P = 0.005) in fish of nutritional history V compared to M (averaged over the three families). Besides the effects on feed intake, V-fish utilized diet V more efficiently than M-fish, as reflected by the on average 18% higher feed efficiency (P = 0.003). We noted a significant family effect for the above parameters (P<0.001), but the nutritional history effect was consistent for all three families (no interaction effect, P>0.05). In summary, our study shows that an early short-term exposure of rainbow trout fry to a plant-based diet improves acceptance and utilization of the same diet when given at later life stages. This positive response is encouraging as a potential strategy to improve the use of plant-based feed in fish, of interest in the field of fish farming and animal nutrition in general. Future work needs to determine the persistency of this positive early feeding effect and the underlying mechanisms. [ABSTRACT FROM AUTHOR]

Compromisation of food intake when confronted with diets deficient in essential amino acids is a common response of fish and other animals, but the underlying physiological factors are poorly understood. We hypothesize that oxygen consumption of fish is a possible physiological factor constraining food intake. To verify, we assessed the food intake and oxygen consumption of rainbow trout fed to satiation with diets which differed in essential amino acid (methionine and lysine) compositions: a balanced vs. an imbalanced amino acid diet. Both diets were tested at two water oxygen levels: hypoxia vs. normoxia. Trout consumed 29% less food under hypoxia compared to normoxia (p<0.001). Under both hypoxia and normoxia trout significantly reduced food intake by 11% and 16% respectively when fed the imbalanced compared to the balanced amino acid diet. Oxygen consumption of the trout per unit body mass remained identical for both diet groups not only under hypoxia but also under normoxia (p>0.05). This difference in food intake between diets under normoxia together with the identical oxygen consumption supports the hypothesis that food intake in fish can be constrained by a set-point value of oxygen consumption, as seen here on a six-week time scale. [ABSTRACT FROM AUTHOR]

This study investigated the hypothesis that the voluntary feed intake in fish is regulated by diet-induced differences in oxygen use. Four diets were prepared with a similar digestible protein:digestible energy ratio (18 mg/kJ), but which differed in the composition of nonprotein energy source. This replacement of fat (F) by starch (S) was intended to create a diet-induced difference in oxygen use (per unit of feed): diets F30-S70, FS0-S50, F65-S35, and F80-S20 with digestible fat providing 28, 49, 65, and 81% of the nonprotein digestible energy (NPDE), respectively. Each diet was fed to satiation to triplicate groups of 20 rainbow trout for 6 wk. As expected, diet-induced oxygen use decreased linearly (R² = 0.89; P < 0.001) with increasing NPDE as fat. The digestible and metabolizable energy intakes of trout slightly increased with increasing NPDE as fat (i.e., decreasing starch content) (R² = 0.30, P = 0.08; and R² = 0.34, P = 0.05, respectively). Oxygen consumption of trout fed to satiation declined with increasing dietary NPDE as fat (R² = 0.48; P = 0.01). The inverse relation between digestible energy intake of trout and the diet-induced oxygen use (R² = 0.33; P = 0.05) suggests a possible role of diet-induced oxygen use in feed intake regulation as shown by the replacement of dietary fat by starch. [ABSTRACT FROM AUTHOR]

Mucins are O-glycosylated glycoproteins present on the apex of all wet-surfaced epithelia with a well-defined expression pattern, which is disrupted in response to a wide range of injuries or challenges. The aim of this study was to identify mucin gene sequences of gilthead sea bream (GSB), to determine its pattern of distribution in fish tissues and to analyse their transcriptional regulation by dietary and pathogenic factors. Exhaustive search of fish mucins was done in GSB after de novo assembly of next-generation sequencing data hosted in the IATS transcriptome database (www.nutrigroup-iats.org/seabreamdb). Six sequences, three categorized as putative membrane-bound mucins and three putative secreted-gel forming mucins, were identified. The transcriptional tissue screening revealed that Muc18 was the predominant mucin in skin, gills and stomach of GSB. In contrast, Muc19 was mostly found in the oesophagus and Muc13 was along the entire intestinal tract, although the posterior intestine exhibited a differential pattern with a high expression of an isoform that does not share a clear orthologous in mammals. This mucin was annotated as intestinal mucin (I-Muc). Its RNA expression was highly regulated by the nutritional background, whereas the other mucins, including Muc2 and Muc2-like, were expressed more constitutively and did not respond to high replacement of fish oil (FO) by vegetable oils (VO) in plant protein-based diets. After challenge with the intestinal parasite Enteromyxum leei, the expression of a number of mucins was decreased mainly in the posterior intestine of infected fish. But, interestingly, the highest down-regulation was observed for the I-Muc. Overall, the magnitude of the changes reflected the intensity and progression of the infection, making mucins and I-Muc, in particular, reliable markers of prognostic and diagnostic value of fish intestinal health. [ABSTRACT FROM AUTHOR]

Abstract: This study examines how dietary macronutrient-induced changes in voluntary food intake (FI) relate to changes in markers of hepatic oxidative metabolism and in the expression of FI regulatory neuropeptides in a teleost model, the rainbow trout. Rainbow trout were fed for 6weeks with one of four iso-energetic diets (2×2 factorial design), containing either a high (HP, ~500g·kg−1 DM) or a low (LP, ~250g·kg−1 DM) protein level (PL) with, at each PL, fat (diets HP-F and LP-F) being substituted by an iso-energetic amount of gelatinized corn starch (diets HP-St and LP-St) as non-protein energy source (ES). Irrespective of the dietary PL, FI (g·kg−0.8·d−1) and digestible energy intake (DEI, kJ·kg−0.8·d−1) were significantly (P<0.05) reduced by the iso-energetic replacement of fat by starch as non-protein ES. Interestingly, trout fed these St-diets had higher gene expression of markers of hepatic oxidative phosphorylation (OxPhos), i.e. ubiquinol-cytochrome c reductase subunit 2 (UCR2) and cytochrome oxidase subunit 4 (COX4) and of aerobic oxidative capacity (CS, citrate synthase), which paralleled glucokinase (GK) transcription. This positive relation suggests that glucose phosphorylation and markers of mitochondrial OxPhos are linked at the hepatic level and possibly triggered the observed reduction in FI. Moreover, trout displaying the reduced FI had higher cocaine amphetamine regulator transcript (CART) mRNA in hypothalamus, whereas neuropeptide Y (NPY) mRNA did not follow the macronutrient-induced changes in FI. Further studies are needed to unravel the mechanisms by which diet-induced changes in hepatic metabolism inform central feeding centers involved in the regulation of FI in fish. [Copyright &y& Elsevier]

The hypothesis was tested that fish fed to satiation with iso-energetic diets differing in macronutrient composition will have different digestible energy intakes (DEI) but similar total heat production. Four iso-energetic diets (2x2 factorial design) were formulated having a contrast in i) the ratio of protein to energy (P/E): high (HP/E) vs. low (LP/E) and ii) the type of nonprotein energy (NPE) source: fat vs. carbohydrate which were iso-energetically exchanged. Triplicate groups (35 fish/tank) of rainbow trout were hand-fed each diet twice daily to satiation for 6 weeks under non-limiting water oxygen conditions. Feed intake (FI), DEI (kJ kg-0.8 d-1) and growth (g kg-0.8 d-1) of trout were affected by the interaction between P/E ratio and NPE source of the diet (P<0.05). Regardless of dietary P/E ratio, the inclusion of carbohydrate compared to fat as main NPE source reduced DEI and growth of trout by ∼20%. The diet-induced differences in FI and DEI show that trout did not compensate for the dietary differences in digestible energy or digestible protein contents. Further, changes in body fat store and plasma glucose did not seem to exert a homeostatic feedback control on DEI. Independent of the diet composition, heat production of trout did not differ (P>0.05). Our data suggest that the control of DEI in trout might be a function of heat production, which in turn might reflect a physiological limit related with oxidative metabolism. [ABSTRACT FROM AUTHOR]

Background: Studies conducted with gilthead sea bream (Sparus aurata L.) have determined the maximum dietary replacement of fish meal and oil without compromising growth or product quality. The present study aimed to analyze the effect of the nutritional background on fish health and fish fed plant protein-based diets with fish oil (FO diet) or a blend of vegetable oils (66VO diet) were exposed for 102 days to the intestinal myxosporean parasite Enteromyxum leei, and the intestine transcriptome was analyzed with a customized oligo-microarray of 7,500 annotated genes. Results: Infection prevalence was high and similar in the two diet groups, but the outcome of the disease was more pronounced in fish fed the 66VO diet. No differences were found in the transcriptome of both diet control groups, whereas the number of differentially expressed genes in infected groups was considerable. K-means clustering of these differentially expressed genes identified four expression patterns that reflected the progression of the disease with the magnitude of the fold-change being higher in infected 66VO fish. A positive correlation was found between the time of infection and the magnitude of the transcriptional change within the 66VO group, being higher in early infected animals. Within this diet group, a strong up-regulation of many components of the immune specific response was evidenced, whereas other genes related to complement response and xenobiotic metabolism were down-regulated. Conclusions: The high replacement of fish oil by vegetable oils in practical fish feeds did not modify the intestine transcriptome of gilthead sea bream, but important changes were apparent when fish were exposed to the myxosporean E. leei. The detected changes were mostly a consequence rather than a cause of the different disease progression in the two diet groups. Hence, the developed microarray constitutes an excellent diagnostic tool to address changes associated with the action of intestinal pathogens, but lacks a prognostic value to predict in advance the different susceptibility of growing fish to the current pathogen. [ABSTRACT FROM AUTHOR]

Abstract: Two trials with juvenile black tiger shrimp (Penaeus monodon) were undertaken to study the effects of replacing fishmeal by different levels of plant proteins on growth performances and nutrient utilisation of shrimp in semi-intensive conditions (Expt. 1) and on the availability of dietary nitrogen (N) and amino acids (Expt. 2). Five isoproteic diets (on crude protein basis) were formulated to contain 34, 24, 16, 8, or 0% fishmeal, with fishmeal being replaced by a mixture of plant protein (corn gluten meal, wheat gluten, and rapeseed meal). In Expt. 1, the shrimp (initial body weight, IBW 1.5±0.1g) were reared in earthen ponds for 144days and fed one of the experimental diets. Apparent digestibility of nutrients and AA were assessed in Expt. 2, using 150L tanks and shrimp of 12.8±0.4g IBW. After 144days in grow-out ponds, shrimp fed the diet with 24% of fishmeal had similar growth as those fed the control diet containing 34% fishmeal (0% replacement). When 50% or more of the fishmeal were replaced, weight gain as well as N and energy gains significantly decreased. Digestibility of dry matter, protein and energy was also significantly lower in all fishmeal-replaced diets. In particular, leucine digestibility decreased by 26% at 100% replacement, which was significantly correlated to an increased incorporation of corn gluten meal. Our data confirm the need to improve our knowledge on AA availability and raw material quality in order to improve fishmeal replacement in P. monodon diets. [Copyright &y& Elsevier]

Abstract: The environmental consequences of replacing fish meal and fish oil with plant-based sources in salmonid feeds were investigated using Life Cycle Assessment (LCA). Two scenarios of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) farming were compared. The first scenario used a Standard Diet (STD) with high levels of fish meal and fish oil, and the second a Low Marine-Fishery-Capture Diet (LFD) in which fish meal and fish oil were replaced by plant protein and oil sources. Environmental impacts assessed were: acidification, eutrophication, climate change, terrestrial ecotoxicity, net primary production use (biotic resource use), water use, land occupation and total cumulative energy demand. The assessment confirmed the substantial contribution of feed to the environmental burdens of fish production and the LFD scenario led to a significant decrease in biotic resource use compared to the STD scenario with the same total energy demand. Environmental impacts of feeds depended highly on the geographic origins of feed ingredients from fishery (e.g., fish oil from Norway or Peru) and from terrestrial agricultural crop species (e.g., palm oil or rapeseed oil). This study demonstrated the importance of a multicriteria method to give stakeholders the most accurate information on the potential consequences of replacing fishery products with plant-based sources in aquafeeds. [Copyright &y& Elsevier]

Fish intestinal microbiota changes with the diet and this effect is of particular interest considering the increasing substitution of fish meal by plant protein sources. The objective of this work was to study the effects of partial substitution of fish meal with lupin and rapeseed meals on gut microbiota of the gilthead sea bream ( Sparus aurata) and in goldfish ( Carassius auratus). Faecal, gastrointestinal and intestinal contents were characterized using culture-based and molecular methods. Vibrionaceae was high in faeces and in the intestine of sea bream, while a more diverse microbiota was retrieved from the stomach, where Bacillales and Flavobacteriaceae appeared to be influenced by the diet. PCR-denaturing gradient gel electrophoresis profiles revealed a high diversity of the microbiota transiting in the sea bream digestive tract, with a shift between gastric and intestinal communities, especially in the group fed with lupin meal. The goldfish was different, with a predominance of Aeromonas spp., Shewanella putrefaciens and Staphylococcus spp. among the aerotolerant-cultivable bacteria. The culture-independent methods revealed the presence of anaerobes like Cetobacterium somerae, and that of Vibrio spp., likely in a viable, but noncultivable state. There was a trend towards decreasing diversity in goldfish microbiota with the partial substitution by lupin, which seemed to inhibit some taxa. [ABSTRACT FROM AUTHOR]

Abstract: Gilthead sea bream (Sparus aurata) were fed in triplicate groups with a commercial standard diet from the juvenile stage to male–female sex reversal under natural day-length and temperature conditions. Every 3–4months during the two-year production cycle, 9 fish were randomly selected and sampled for flesh composition analyses of total lipid levels and fatty acid (FA) composition. Curvilinear regressions fitting total lipid levels and % FAs in total lipids were made to underline the differential distribution of a given fillet FA within neutral and polar lipid fractions. This dataset along with published results on market-size fish were combined for multilinear regression approaches, with the aim of describing strong relationships (P<0.0001) between fillet FA composition and two independent variables: dietary FA composition and fillet lipid level. For saturated (14:0, 16:0, 18:0) and monounsaturated (16:1n-7, 18:1n-7, 18:1n-9, 20:1n-9) FAs, the overall variance in fillet FA composition is primarily explained by dietary FA composition and secondly by fillet lipid level. This second independent variable also contributes to explain the variations observed in arachidonic acid (20:4n-6) and docosahexaenoic acid (22:6n-3), but a statistically significant contribution is not found for linoleic acid (18:2n-6), linolenic acid (18:3n-3), eicosapentaenoic acid (20:5n-3) and docosapentaenoic acid (22:5n-3). The consistency of these predictive equations in our particular rearing conditions was proved by means of a test validation trial, using fish fed an experimental diet based on plant proteins and fish oil. [Copyright &y& Elsevier]

Abstract: The aim of this work was to underline the physiological role of the antioxidant peroxiredoxin (PRDX) family in gilthead sea bream (Sparus aurata L.), a perciform fish extensively cultured in the Mediterranean area. First, extensive BLAST searches were done on the gilthead sea bream cDNA database of the AQUAMAX European Project (www.sigenae.org/iats), and six contigs were unequivocally identified as PRDX1-6 after sequence completion by RT-PCR. The phylogenetic analysis evidenced three major clades corresponding to PRDX1-4 (true 2-Cyst PRDX subclass), PRDX5 (atypical 2-Cys PRDX subclass) and PRDX6 (1-Cys PRDX subclass) that reflected the present hierarchy of vertebrates. However, the PRDX2 branch of modern fish including gilthead sea bream was related to the monophyletic PRDX1 node rather than to PRDX2 cluster of mammals and primitive fish, which probably denotes the acquisition of novel functions through vertebrate evolution. Transcriptional studies by means of quantitative real-time PCR evidenced a ubiquitous PRDX gene expression that was tissue specific for each PRDX isoform. In a second set of transcriptional studies, liver and head kidney were chosen as target tissues in fish challenged with i) the intestinal parasite Enteromyxum leei, ii) a plant oil (VO) diet with deficiencies in essential fatty acids and iii) prolonged exposure to high-rearing densities. These studies showed that PRDX genes were highly and mostly constitutively expressed in the liver and were not affected by dietary intervention or high density. In contrast, head kidney was highly sensitive to the different experimental challenges: significantly lower values were found for PRDX5 in the three trials, for PRDX6 in parasitized and high density fish and for PRDX1 in parasitized and VO fish. PRDX2, 3 and 5 were decreased only in VO, high density and parasitized animals, respectively. These findings would highlight the role of PRDXs as integrative and highly predictive biomarkers of health and welfare in fish and gilthead sea bream in particular. [Copyright &y& Elsevier]

Most teleost fish are known to require high levels of dietary proteins. Such high-protein intake could have significant effects, particularly on insulin-regulated gene expression. We therefore analyzed the effects of an increase in the ratio of dietary carbohydrates/proteins on the refeeding activation of the Akt-target of rapamycin (TOR) signaling pathways in rainbow trout and the effects on the expression of several genes related to hepatic and muscle metabolism and known to be regulated by insulin, amino acids, and/or glucose. Fish were fed once one of three experimental diets containing high (H), medium (M), or low (L) protein (P) or carbohydrate (C) levels after 48 h of feed deprivation. Activation of the Akt/TOR signaling pathway by refeeding was severely impaired by decreasing the proteins-to-carbohydrates ratio. Similarly, postprandial regulation of several genes related to glucose (Glut4, glucose-6-phosphatase isoform 1), lipid (fatty acid synthase, ATP-citrate lyase, sterol responsive element binding protein, carnitine palmitoyltransferase 1, and 3-hydroxyacyl-CoA dehydrogenase), and amino acid metabolism (serine dehydratase and branched-chain α-keto acid dehydrogenase E2 subunit) only occurred when fish were fed the high-protein diet. On the other hand, diet composition had a low impact on the expression of genes related to muscle protein degradation. Interestingly, glucokinase was the only gene of those monitored whose expression was significantly upregulated by increased carbohydrate intake. In conclusion, this study demonstrated that macro-nutrient composition of the diet strongly affected the insulin/amino acids signaling pathway and expression pattern of genes related to metabolism. [ABSTRACT FROM AUTHOR]

This preliminary study assessed genotype × diet interaction in late growth of European sea bass ( Dicentrarchus labrax) fed with either a fish meal (FM)- or a fish oil (FO)-based diet (M) or an all-plant-based (PB) diet. A total of 550 fish from 224 families were reared together and tagged. DNA was sampled and microsatellites were used to assign parentage. When fish weight was 192 ± 54 g, two tanks were fed with M (FM: 100%; FO: 100%) and two others with PB (FM: 0% and FO: 0%). Body weight (BW), fork length (FL) and fillet lipid content ( CorrFat) were analysed with a linear model and with REML methodology. We observed no significant differences between groups, but a slightly lower ( P=0.03) daily growth coefficient in sea bass fed PB than in those fed M. Heritability estimates of BW differed significantly from zero (PB: 0.37 ± 0.18; M: 0.47 ± 0.24). Sire × diet interactions were significant and genetic correlations ranged between 0.51 and 0.87, showing genotype × diet interaction for BW and CorrFat. For the first time, genetic parameters in the context of total replacement of marine fishery by-products were estimated in European sea bass, showing re-ranking of family performances with extremely contrasted diets. [ABSTRACT FROM AUTHOR]

Abstract: The present study aimed to ascertain the different fatty acid (FA) descriptors linking dietary and muscle FA composition in one-year-old gilthead sea bream. For that purpose, our own published data along with additional data from the present study were compiled and analysed. High linear correlations (r 2 =0.90, P <0.001) between dietary and muscle fatty acid composition were reported for monoenes, C18 polyunsaturated FA (PUFA) and long-chain PUFA (LC-PUFA). Prediction deviations due to changes in muscle fatness were analysed in an independent trial with two different feeding levels (full ration size, 30% restriction ration). Regardless of feeding regimen, predicted values for muscle FA at low concentrations deviated (P <0.001) from observed values, but good predictions with less than 6% deviations were found for abundant fatty acids (16:1n−7, 18:1n−9, 18:2n−6, 18:3n−3, 20:4n−6, 20:5n−3, 22:6n−3). All this highlights the predictable effects of dietary oils in the muscle FA composition of gilthead sea bream, although further research is needed to cover all the range of commercial fish size and for the up-scaling of laboratory results to different fish farming conditions. [ABSTRACT FROM AUTHOR]

Abstract: The aim of the present work was to determine if a practical plant protein-based diet containing vegetable oils (VO) as the major lipid source could alter the disease course when challenged with the myxosporean Enteromyxum leei, a wide-spread parasite in the Mediterranean basin causing heavy economic losses. Gilthead sea bream (Sparus aurata) fed for 9 months either a fish oil (FO) diet or a blend of VOs at 66% of replacement (66VO diet) were challenged by exposure to parasite-contaminated water effluent. All fish were periodically and non-lethally sampled to obtain biometrical data and to know their infection status. After 102 days of exposure, fish were euthanized and haematological, biometrical, histological, immunological, glutathione and anti-oxidant data were obtained from tissue, blood and serum samples. Anorexia appeared in both exposed groups, but feed intake reduction was higher in 66VO fish. The signs of disease (lower growth, condition factor, specific growth rate, haematocrit) as well as the disease course were worse in fish from 66VO group, with a higher prevalence and intensity of infection, a higher percentage of fish harbouring the parasite in the entire intestinal tract, and a faster establishment of the parasite. Parasite intensity of infection was negatively correlated with growth parameters and haematocrit in both groups, and with complement, lysozyme and hepatic total glutathione in 66VO fish. [ABSTRACT FROM AUTHOR]

Abstract: In mammals, new evidence has demonstrated the important role of the autophagic/lysosomal pathway in regulating muscle mass and identified the transcription factor FoxO3 as a key factor of the control of this proteolytic system by inducing several autophagy-related genes. In contrast, the mechanisms responsible for the regulation of autophagy have not been investigated in teleosts, known to exhibit different muscle growth dynamics. The present work aimed to characterize both in vivo and in vitro the transcriptional regulation of several major genes involved in autophagy (LC3B, gabarapl1, atg12l, atg4b) in the white skeletal muscle of rainbow trout. We found that fasting fish for 14days or serum depletion of trout myocytes strongly induces the expression of all studied genes. Our in vitro study on trout myocytes indicated that IGF1 induces FoxO3 phosphorylation but has a low or no effect on autophagy-related gene expression, suggesting a moderate role for this transcription factor on the autophagic/lysosomal pathway in this species. Data reported here show for the first time in a lower vertebrate, the existence and the regulation of several major genes involved in the autophagy, opening a new area of research on the molecular bases of muscle protein degradation in teleosts. [ABSTRACT FROM AUTHOR]

Abstract: This study evaluates the influence of both dietary protein and methionine on amino acid trans- and deamination (alanine aminotransferase, ALAT and glutamate dehydrogenase, GDH) in three tissues (muscle, hepatopancreas, gills) of the marine black tiger shrimp (Penaeus monodon). Shrimp (2.4g) were fed one of the six semi-purified diets containing 14, 34 or 54% crude protein (% dry matter) with two levels of methionine (normal or 30% reduced) for 6weeks. Both ALAT and GDH activities were the highest in the muscle. ALAT activity in muscle significantly decreased when feeding the low vs. high protein diets. Compared to those fed the intermediate protein level, GDH activity in muscle decreased (by 35%) when fed the low and increased (by 26%) when fed the high protein diets (P<0.05). A significant interaction between dietary protein and methionine was observed on GDH activity in gills which, due to the relative methionine deficiency, increased 4-fold at the intermediate protein level. In summary, our results demonstrate for the first time the capacity of up and downregulation of enzyme activity by dietary protein levels in the muscle of P. monodon, and the active role played by branchial tissue in ammoniogenesis in response to a relative indispensable amino acid (methionine) deficiency. [ABSTRACT FROM AUTHOR]