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This study investigated the influence of dietary astaxanthin (AX) on glucose and lipid metabolism in rainbow trout liver. Two iso-nitrogenous and iso-lipidic diets were tested for 12 weeks in rainbow trout with an initial mean weight of 309 g. The S-ASTA diet was supplemented with 100 mg of synthetic AX per kg of feed, whereas the control diet (CTRL) had no AX. Fish fed the S-ASTA diet displayed lower neutral and higher polar lipids in the liver, associated with smaller hepatocytes and lower cytoplasm vacuolization. Dietary AX upregulated adipose triglyceride lipase (atgl), hormone-sensitive lipase (hsl2) and 1,2-diacylglycerol choline phosphotransferase (chpt), and downregulated diacylglycerol acyltransferase (dgat2), suggesting the AX's role in triacylglycerol (TAG) turnover and phospholipid (PL) synthesis. Dietary AX may also affect beta-oxidation with the upregulation of carnitine palmitoyltransferase 1 (cpt1α2). Although hepatic cholesterol levels were not affected, dietary AX increased gene expression of sterol regulatory element-binding protein 2 (srebp2). Dietary AX upregulated the expression of 6-phosphogluconate dehydrogenase (6pgdh) and downregulated pyruvate kinase (pkl). Overall, results suggest that dietary AX modulates the oxidative phase of the pentose phosphate pathway and the last step of glycolysis, affecting TAG turnover, β-oxidation, PL and cholesterol synthesis in rainbow trout liver. [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]

This study evaluated how different forms of selenium (Se) supplementation into rainbow trout broodstock diets modified the one-carbon metabolism of the progeny after the beginning of exogenous feeding and followed by hypoxia challenge. The progeny of three groups of rainbow trout broodstock fed either a control diet (Se level: 0·3 µg/g) or a diet supplemented with inorganic sodium selenite (Se level: 0·6 µg/g) or organic hydroxy-selenomethionine (Se level: 0·6 µg/g) was cross-fed with diets of similar Se composition for 11 weeks. Offspring were sampled either before or after being subjected to an acute hypoxic stress (1·7 mg/l dissolved oxygen) for 30 min. In normoxic fry, parental Se supplementation allowed higher glutathione levels compared with fry originating from parents fed the control diet. Parental hydroxy-selenomethionine treatment also increased cysteine and cysteinyl–glycine concentrations in fry. Dietary Se supplementation decreased glutamate–cysteine ligase (cgl) mRNA levels. Hydroxy-selenomethionine feeding also lowered the levels of some essential free amino acids in muscle tissue. Supplementation of organic Se to parents and fry reduced betaine-homocysteine S-methyltransferase (bhmt) expression in fry. The hypoxic stress decreased whole-body homocysteine, cysteine, cysteinyl-glycine and glutathione levels. Together with the higher mRNA levels of cystathionine beta-synthase (cbs), a transsulphuration enzyme, this suggests that under hypoxia, glutathione synthesis through transsulphuration might have been impaired by depletion of a glutathione precursor. In stressed fry, S-adenosylmethionine levels were significantly decreased, but S-adenosylhomocysteine remained stable. Decreased bhmt and adenosylmethionine decarboxylase 1a (amd1a) mRNA levels in stressed fry suggest a nutritional programming by parental Se also on methionine metabolism of rainbow trout. [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]

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]

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]

The rainbow trout (Oncorhynchus mykiss) exhibits high dietary amino acid requirements and an apparent inefficiency to use dietary carbohydrates. Using this species, we investigated the metabolic consequences of long-term high carbohydrates/low protein feeding. Fish were fed two experimental diets containing either 20 % carbohydrates/50 % proteins (C20P50), or high levels of carbohydrates at the expense of proteins (35 % carbohydrates/35 % proteins – C35P35). The expression of genes related to hepatic and muscle glycolysis (glucokinase (GK), pyruvate kinase and hexokinase) illustrates the poor utilisation of carbohydrates irrespective of their dietary levels. The increased postprandial GK activity and the absence of inhibition of the gluconeogenic enzyme glucose-6-phosphatase activity support the hypothesis of the existence of a futile cycle around glucose phosphorylation extending postprandial hyperglycaemia. After 9 weeks of feeding, the C35P35-fed trout displayed lower body weight and feed efficiency and reduced protein and fat gains than those fed C20P50. The reduced activation of eukaryotic translation initiation factor 4-E binding protein 1 (4E-BP1) in the muscle in this C35P35 group suggests a reduction in protein synthesis, possibly contributing to the reduction in N gain. An increase in the dietary carbohydrate:protein ratio decreased the expression of genes involved in amino acid catabolism (serine dehydratase and branched-chain α-keto acid dehydrogenase E1α and E1β), and increased that of carnitine palmitoyltransferase 1, suggesting a higher reliance on lipids as energy source in fish fed high-carbohydrate and low-protein diets. This probably also contributes to the lower fat gain. Together, these results show that different metabolic pathways are affected by a high-carbohydrate/low-protein diet in rainbow trout. [ABSTRACT FROM PUBLISHER]

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]

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]

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]

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]

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]

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]

Rainbow trout have a limited ability to use dietary carbohydrates efficiently and are considered to be glucose intolerant. Administration of carbohydrates results in persistent hyperglycemia and impairs post-prandial down regulation of gluconeogenesis despite normal insulin secretion. Since gluconeogenic genes are mainly under insulin control, we put forward the hypothesis that the transcriptional function of insulin as a whole may be impaired in the trout liver. In order to test this hypothesis, we performed intraperitoneal administration of bovine insulin to fasted rainbow trout and also subjected rainbow trout primary hepatocytes to insulin and/or glucose stimulation. We demonstrate that insulin was able to activate Akt, a key element in the insulin signaling pathway, and to regulate hepatic metabolism-related target genes both in vivo and in vitro. In the same way as in mammals, insulin decreased mRNA expression of gluconeogenic genes, including glucose 6-phosphatase (G6Pase), fructose 1 ,6-bisphosphatase (FBPase) and phosphoenolpyruvate carboxykinase (PEPCK). Insulin also limited the expression of carnitine palmitoyltransferase 1 (CPTI), a limiting enzyme of fatty acid ~3-oxidation. In vitro studies revealed that, as in mammals, glucose is an important regulator of some insulin target genes such as the glycolytic enzyme pyruvate kinase (PK) and the lipogenic enzyme fatty acid synthase (FAS). Interestingly, glucose also stimulates expression of glucokinase (GK), which has no equivalent in mammals. This study demonstrates that insulin possesses the intrinsic ability to regulate hepatic gene expression in rainbow trout, suggesting that other hormonal or metabolic factors may counteract some of the post-prandial actions of insulin. [ABSTRACT FROM AUTHOR]

In mammals, feeding promotes protein accretion in skeletal muscle through a stimulation of the insulin- and amino acid-sensitive mammalian target of rapamycin (mTOR) signaling pathway, leading to the induction of mRNA translation. The purpose of the present study was to characterize both in vivo and in vitro the activation of several major kinases involved in the mTOR pathway in the muscle of the carnivorous rainbow trout. Our results showed that meal feeding enhanced the phosphorylation of the target of rapamycin (TOR), PKB, p70 S6 kinase, and eIF4E-binding protein-i, suggesting that the mechanisms involved in the regulation of mRNA translation are well conserved between lower and higher vertebrates. Our in vitro studies on primary culture of trout muscle cells indicate that insulin and amino acids regulate TOR signaling and thus may be involved in meal feeding effect in this species as in mammals. In conclusion, we report here for the first time in a fish species, the existence and the nutritional regulation of several major kinases involved in the TOR pathway, opening a new area of research on the molecular bases of amino acid utilization in teleosts. [ABSTRACT FROM AUTHOR]

The objective of the present study was to characterise some important antioxidant enzymes and their relationships with retinoids and lipid peroxidation during rainbow trout (Oncorhynchus mykiss) early development. Eggs were incubated at 7°C until the swim-up stage whereupon fry were fed two semi-purified diets with 0 % (CO) and 8 % (OX) oxidised lipid respectively for 2 months at 17°C. The activities and gene expression of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) were determined as well as the levels of retinoids, F2-isoprostanes and lipid-soluble fluorescent products (LSFP) at various developmental stages. Only SOD had a detectable activity in embryos which increased during development and was linked with an increase of mitochondrial (SOD2) and cytosolic (SOD1) gene expression. SOD1 and SOD2 mRNA were more abundant in fry fed OX than in fry fed CO. CAT activity and gene expression also increased during development and were higher in fry fed OX compared with fry fed CO. Activity of Se-dependent GPX (Se-GPX) increased during development. The gene expression of cytosolic Se-GPX (GPX1) increased from hatching to 2-month-fed fry. Both phospholipid-hydroperoxide GPX and GPX1 genes were more expressed in fry fed OX than in fry fed CO. Retinoids decreased during development and, by 2 months, were lowered in fry fed OX compared with those fed CO. The levels of LSFP were higher in fry fed OX compared with fry fed CO. The present study demonstrates that antioxidant defence systems are active all through the development of rainbow trout and modulated by feeding oxidised lipid. [ABSTRACT FROM PUBLISHER]

Rainbow trout is unable to utilize high levels of dietary carbohydrates and experiences hyperglycemia after consumption of carbohydrate-rich meals. Carbohydrates stimulate hepatic glycolytic activity, but gene expression of the rate-limiting gluconeogenic enzymes glucose-6-phosphatase (G6Pase), fructose-1,6-bisphosphatase (FBPase) and phosphoenolpyruvate carboxykinase (PEPCK) remains high. Although there is significant mRNA expression and activity of gluconeogenic enzymes in trout intestine and kidney, the regulation of these enzymes by diet is not known. We tested the hypothesis that dietary carbohydrate modulates intestinal and renal G6Pase, FBPase and PEPCK. Fish were either fasted or fed isocaloric carbohydrate-free (CF) or high carbohydrate (HC) diets for 14 days. As expected, fish fed HC exhibited postprandial hyperglycemia and enhanced levels of hepatic glucokinase mRNA and activity. Dietary carbohydrates had no significant effect on the expression and activity of PEPCK, FBPase and G6Pase in all three organs. In contrast, fasting enhanced the activity, but not the mRNA expression of both hepatic and intestinal PEPCK, as well as intestinal FBPase. Therefore, the activity of rate-limiting gluconeogenic enzymes in trout can be modified by fasting, but not by the carbohydrate content of the diet, potentially causing hyperglycemia when fed high levels of dietary carbohydrates. In this species consuming low carbohydrate diets at infrequent intervals in the wild, fasting-induced increases in hepatic and intestinal gluconeogenic enzyme activities may be a key adaptation to prevent perturbations in blood glucose during food deprivation. [ABSTRACT FROM AUTHOR]

In mammals, the ubiquitin-proteasome proteolytic pathway is a major route of protein degradation and has been shown to be regulated by the feeding status via the protein kinase B (PKB)-Forkehead box-O transcription factor signaling pathway-mediated transcription regulation of atrophy-related ubiquitin ligases, atrogin1 and muscle RING finger 1. In contrast, in rainbow trout (Oncorhynchus mykiss), the activity of the proteasome in muscle was not affected during starvation-induced muscle degradation. The aim of this study was therefore to explore the molecular basis for this lack of induction of this proteolytic route during starvation. In this study, rainbow trout were food deprived for 7 and 14 d, refed ad libitum, and the effect of the nutritional status was assessed on the different steps involved in the regulation of the ubiquitin-proteasome system in muscle. We observed that starvation reduced the phosphorylation of PKB and enhanced the expression of atrogin1 in muscle, whereas refeeding led to the opposite effects. The level of polyubiquitinated proteins in muscle increased to over 2 times the initial value on d 0 after 14 d of starvation and decreased significantly at 12 h after refeeding, but there were no major changes in the activity of the main proteasomal peptidases (chymotrypsin-like and trypsin-like). Altogether, these results indicate that in rainbow trout muscle, the polyubiquitination step of the ubiquitin-proteasome route is regulated by the feeding status similarly to what is observed in mammals. [ABSTRACT FROM AUTHOR]

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]

In order to investigate the effects of dietary lipid sources on mechanisms involved in lipid deposition, two groups of rainbow trout were fed from first-feeding to the commercial size of 1kg (for 62 weeks) with two diets differing only by lipid source: 100% fish oil or 100% blend of vegetable oils (55% rapeseed oil, 30% palm oil, 15% linseed oil). The activities and levels of gene expression of lipogenic enzymes (fatty acid synthetase, glucose-6-phosphate dehydrogenase and malic enzyme) in liver and of lipoprotein lipase in perivisceral adipose tissue, white muscle and liver were determined. Transport of lipid was studied by determining lipid composition of plasma and lipoprotein classes. We also examined the clearance of LDL by assaying the level of LDL receptor gene expression in several tissues. Total replacement of dietary fish oil by the blend of vegetable oils did not affect growth of rainbow trout and did not modify muscle lipid content. Hepatic lipogenesis and lipid uptake in perivisceral adipose tissue, white muscle and liver were also not modified by dietary treatments. Diets containing the blend of vegetable oils induced a decrease in plasma cholesterol and LDL. In trout fed the vegetable oils diet, expression of LDL receptor gene in the liver was down-regulated. [ABSTRACT FROM PUBLISHER]

The activity of the somatotropic axis was analysed in juvenile rainbow trout (Oncorhynchus mykiss) fed either a fishmeal-based diet (FM) or graded levels of plant proteins to replace 50 % (PP50 diet), 75 % (PP75 diet) or 100 % (PP100 diet) of the fishmeal protein. For this purpose, partial cloning and sequencing of the gene encoding rainbow trout growth hormone receptor (GHR) was first accomplished by RT-PCR, using degenerate primers based on the sequences of non-salmonid fish GHR. Growth rates and energy retention were lowered by the PP75 and PP100 diets and a concurrent and progressive increase in plasma levels of growth hormone (GH) was found. However, no changes in hepatic GH binding and total plasma insulin-like growth factor (IGF)-I levels were observed among the four experimental groups. This fact agrees with the lack of changes in hepatic measurements of GHR and IGF-I transcripts. No consistent changes in IGF transcripts were found in peri-visceral adipose tissue and skeletal muscle, but GHR mRNA was up-regulated in the peri-visceral adipose tissue of fish fed the PP75 and PP100 diets, which would favour the lipolytic action of GH. Two specific bands (47 and 33 kDa) of IGF-binding proteins were found in the plasma of all analysed fish, but the sum of the two integrated areas increased progressively with plant protein supply, which might reflect a reduced free IGF availability. Therefore, in our experimental model, the growth impairment could be due, at least in part, to a lowered availability of biologically active IGF (free IGF fraction) rather than to liver GH desensitization or defect in IGF synthesis and release at the systemic and/or paracrine???autocrine level. [ABSTRACT FROM PUBLISHER]

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]

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]

Acetyl-CoA carboxylase (ACoAC) catalyses the carboxylation of acetyl-CoA into malonyl-CoA. This product plays a pivotal role in the regulation of energy metabolism since it is both a substrate for fatty acid synthesis and an inhibitor of the oxidative pathway. The present study was initiated to analyse the modulation of ACoAC activity in liver and selected extrahepatic tissues of rainbow trout (Oncorhynchus mykiss) by dietary changes as a contribution to the understanding of the nutritional control of lipid metabolism in fish. Short-term effects of food intake were studied by measuring ACoAC activity in the liver and dorsal white muscle at different time intervals after a meal. Only slight variations were observed in the muscle during the period 2???72 h after the meal. The long-term effects of an increase in dietary lipids or carbohydrates levels were examined by measuring ACoAC activity in the liver, adipose tissue, intestine, kidney, red muscle, dorsal and ventral white muscles of trout after 3 months of feeding with different diets. ACoAC activity is stimulated by a high-digestible starch diet in the abdominal adipose tissue and the white muscle. A high-lipid diet decreases ACoAC activity in the liver and the intestine, but not in other tissues. Contrary to mammals, a rapid adaptation of ACoAC activity to food supply is not effective in rainbow trout. However, a long-term nutritional control of ACoAC activity does occur in this species, but the target tissue differs with the predominant non-protein energy sources in the diet. The present results suggest the potential existence of two ACoAC isoforms with different tissue distribution as has been observed in mammals and birds. [ABSTRACT FROM PUBLISHER]

Two rapeseed (Brassica napus) meals, RM1 and RM2, with two levels of glucosinolates (GLS; 5 and 41 μmol/g DM respectively) were incorporated at the levels of 300 and 500 g/kg of the diets of juvenile rainbow trout (Oncorhynchus mykiss) in replacement of fish meal, and compared with a fish-meal-based diet. A decrease in the digestibility of the DM, protein, gross energy and P was observed with high-rapeseed meal (RM) incorporation. In trout fed on RM-based diets, growth performance was reduced even after only 3 weeks of feeding. Feed efficiency was adversely affected by RM and GLS intake. Protein and energy retention coefficients were significantly lower in fish fed on the diet containing the higher level of GLS. P retention was significantly lower with all the RM-based diets than with the fish-meal diet. Irrespective of the degree of growth inhibition, fish fed on RM-based diets exhibited similar typical features of hypothyroid condition due to GLS intake, expressed by lower plasma levels of triiodothyronine and especially thyroxine and a hyperactivity of the thyroid follicles. This hypothyroidal condition led to a strong adjustment of the deiodinase activities in the liver, the kidney and the brain. A significant increase of the outer ring deiodinase activities (deiodinases type I and II respectively) and a decrease of the inner ring deiodinase activity (deiodinase type III) were observed. It is concluded that the observed growth depression could be attributed to the concomitant presence of GLS, depressing the thyroid function, and of other antinutritional factors affecting digestibility and the metabolic utilization of dietary nutrients and energy. [ABSTRACT FROM PUBLISHER]

Selenium is an essential micronutrient and its metabolism is closely linked to the methionine cycle and transsulfuration pathway. The present study evaluated the effect of two different selenium supplements in the diet of rainbow trout (Onchorhynchus mykiss) broodstock on the one-carbon metabolism and the hepatic DNA methylation pattern in the progeny. Offspring of three parental groups of rainbow trout, fed either a control diet (NC, basal Se level: 0.3 mg/kg) or a diet supplemented with sodium selenite (SS, 0.8 mg Se/kg) or hydroxy-selenomethionine (SO, 0.7 mg Se/kg), were collected at swim-up fry stage. Our findings suggest that parental selenium nutrition impacted the methionine cycle with lower free methionine and S-adenosylmethionine (SAM) and higher methionine synthase (mtr) mRNA levels in both selenium-supplemented treatments. DNA methylation profiling by reduced representation bisulfite sequencing (RRBS) identified differentially methylated cytosines (DMCs) in offspring livers. These DMCs were related to 6535 differentially methylated genes in SS:NC, 6890 in SO:NC and 7428 in SO:SS, respectively. Genes with the highest methylation difference relate, among others, to the neuronal or signal transmitting and immune system which represent potential targets for future studies. [ABSTRACT FROM AUTHOR]

Glucose intolerance and limited efficiency in dietary carbohydrates use are characteristic features of carnivorous fish. The absence of post-prandial down-regulation of gluconeogenic gene expressions in rainbow trout suggests an insulin resistance state as observed in human patients with non-insulin-dependent diabetes mellitus. In order to investigate the ability of insulin to control hepatic gene expression in rainbow trout, we performed intra-peritoneal injection of bovine insulin to 24 hours-fasted rainbow trout. We demonstrated that insulin was able in vivo to activate its signaling pathway as indicated by the activation of the protein kinase B and to modulate expression of several genes including glucose-6-phosphatase and phosphoenolpyrnvate carboxykinase, carnitine palmitoyltransferase 1 and PPAR gamma co-activator 1 alpha. These results demonstrated the intrinsic capacity of insulin to regulate gene expression in rainbow trout liver. Consequently, the absence of post-prandial regulation of hepatic gluconeogenesis in trout cannot be directly attributed to default in insulin signaling. Since amino acids have recently been proposed as potent down regulators of insulin signaling in mammals we hypothesize that the ability of insulin to control expression of metabolic genes could be disrupted in carnivorous fish by the high dietary protein levels. This hypothesis is under investigation. [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]

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]

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]