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The effects of replacing fishmeal and fish oil with a plant-based diet were studied in juvenile (10g) and ongrowing (250-350g) rainbow trout from first-feeding. Feed-related differences in the intestinal and hepatic transcriptome were examined in juveniles after 7 months of feeding at 7°C. Based on microarray results obtained for juveniles, the expression of selected genes related to lipid, cholesterol and energy metabolisms, was assessed by RT-qPCR in ongrowing trout after 6 additional months of feeding at 17°C. Plasma glucose and cholesterol, lipid content and fatty acid profile of whole body were analyzed at both stages. After 7 months at 7°C, all juveniles reached the same body weight (10g), while at 13 months ongrowing fish fed the totally plant-based diet exhibited lower body weight (234 vs 330-337g). Body lipid content was higher in juveniles fed the totally plant-based diet (13.2 vs 9.4–9.9%), and plasma cholesterol was about 2-times lower in trout fed the plant-based diets at both stages. Fatty acid profile mirrored that of the respective diet, with low proportions of long-chain n-3 polyunsaturated fatty acids in fish fed plant-based diets. Genes involved in protein catabolism, carbohydrate metabolism and trafficking were down-regulated in the intestines of juveniles fed the plant-based diets. This was not true for ongrowing fish. Genes involved in lipid and cholesterol metabolisms were up-regulated in the livers of fish fed plant-based diets for both stages. In this study, feeding trout a totally plant-based diet from first-feeding affect a relatively low proportion of metabolism-related genes. In the longer term, when fish were reared at a higher temperature, only some of these changes were maintained (i.e. up-regulation of lipid/cholesterol metabolism). Although the plant-based diets tested in this study had no major deficiencies, small adjustments in the feed-formula are needed to further optimize growth performance while sparing marine resources. [ABSTRACT FROM AUTHOR]

In the context of limited marine resources, the exponential growth of aquaculture requires the substitution of fish oil and fishmeal, the traditional components of fish feeds by terrestrial plant ingredients. High levels of such substitution are known to negatively impact fish performance such as growth and survival in rainbow trout (Oncorhynchus mykiss) as in other salmonids. In this respect, genetic selection is a key enabler for improving those performances and hence for the further sustainable development of aquaculture. We selected a rainbow trout line over three generations for its ability to survive and grow on a 100% plant-based diet devoid of both fish oil and fishmeal (V diet) from the very first meal. In the present study, we compared the control line and the selected line after 3 generations of selection, both fed either the V diet or a marine resources-based diet (M diet). The objective of the study was to assess the efficiency of selection and the consequences on various correlated nutritional traits: feed intake, feed efficiency, digestibility, composition of whole fish, nutrient retention and fatty acid (FA) profile. We demonstrated that the genetic variability present in our rainbow trout population can be selected to improve survival and growth. The major result of the study is that after only three generations of selection, selected fish fed the V diet grew at the same rate as the control line fed the M diet, whilst the relative reduction of body weight was 36.8% before the selection. This enhanced performance on the V diet seems to be mostly linked to a higher feed intake for the selected fish. [ABSTRACT FROM AUTHOR]

Two experimental rainbow trout lines developed through divergent selection for low (Lean ‘L’ line) or high (Fat ‘F’ line) muscle fat content were used as models to study the genetic determinism of fat depots. Previous nutritional studies suggested that the F line had a better capability to use glucose than the L line during feeding trials. Based on that, we put forward the hypothesis that F line has a greater metabolic ability to clear a glucose load effectively, compared to L line. In order to test this hypothesis, 250 mg/kg glucose was intraperitoneally injected to the two rainbow trout lines fasted for 48 h. Hyperglycemia was observed after glucose treatment in both lines without affecting the phosphorylation of AMPK (cellular energy sensor) and Akt-TOR (insulin signaling) components. Liver glucokinase and glucose-6-phosphate dehydrogenase expression levels were increased by glucose, whereas mRNA levels of β-oxidation enzymes (CPT1a, CPT1b, HOAD and ACO) were down-regulated in the white skeletal muscle of both lines. Regarding the genotype effect, concordant with normoglycemia at 12 h after glucose treatment, higher muscle glycogen was found in F line compared to L line which exhibited hyperglycemia. Moreover, mRNA levels of hepatic glycolytic enzymes (GK, 6PFK and PK), gluconeogenic enzyme PEPCK and muscle fatty acid oxidation enzymes (CPT1a, CPT1b and HOAD) were concurrently higher in the F line. Overall, these findings suggest that F line may have a better ability to maintain glucose homeostasis than L line. [ABSTRACT FROM AUTHOR]

The present study investigated the effect of dietary carbohydrates on metabolism, with special focus on fatty acid bioconversion and flesh lipid composition in two rainbow trout lines divergently selected for muscle lipid content and fed with vegetable oils. These lines were chosen based on previously demonstrated potential differences in LC-PUFA synthesis and carbohydrate utilization. Applying a factorial study design, juvenile trout from the lean (L) and the fat (F) line were fed vegetable oil based diets with or without gelatinised starch (17.1%) for 12 weeks. Blood, liver, muscle, intestine and adipose tissue were sampled after the last meal. Feed intake and growth was higher in the L line than the F line, irrespective of the diet. Moderate postprandial hyperglycemia, strong induction of hepatic glucokinase and repressed glucose-6-phosphatase transcripts confirmed the metabolic response of both lines to carbohydrate intake. Further at the transcriptional level, dietary carbohydrate in the presence of n-3 LC-PUFA deficient vegetable oils enhanced intestinal chylomicron assembly, disturbed hepatic lipid metabolism and importantly elicited a higher response of key desaturase and elongase enzymes in the liver and intestine that endorsed our hypothesis. PPARγ was identified as the factor mediating this dietary regulation of fatty acid bioconversion enzymes in the liver. However, these molecular changes were not sufficient to modify the fatty acid composition of muscle or liver. Concerning the genotype effect, there was no evidence of substantial genotypic difference in lipid metabolism, LC-PUFA synthesis and flesh fatty acid profile when fed with vegetable oils. The minor reduction in plasma glucose and triglyceride levels in the F line was linked to potentially higher glucose and lipid uptake in the muscle. Overall, these data emphasize the importance of dietary macro-nutrient interface in evolving fish nutrition strategies. [ABSTRACT FROM AUTHOR]

Abstract: This study examined the genetic variability and genotype×diet interactions during early growth (initial mean body weight 1.2g) among seven heterozygous clones of rainbow trout, Oncorhynchus mykiss. The clones were hand-fed a diet containing either fishmeal or plant proteins during a 49-day trial divided into two periods (P1, 26days, and P2, 23days). Weight, variation of weight within clone, feed intake, feed efficiency and mortality were calculated for both periods. There was a highly significant effect of diet and of clone for all traits at both periods, except for feed efficiency and mortality at P1. Highly significant interactions between diet and clone were also recorded for all these traits, except for mortality at P1. The occurrence of genotype×diet interactions when feeding juvenile rainbow trout with an all plant-protein diet indicates that a highly performing genotype on a fishmeal diet may perform poorly when fed a plant-protein diet. Interactions were found for the two major determinants of growth, i.e. feed intake and feed efficiency, showing that the dietary response differs according to the genotype. Monitoring of the within-clone variability of weight showed that a plant-based diet is likely to enhance the overall phenotypic variance in a population, whatever its initial genetic variability. [Copyright &y& Elsevier]

Rainbow trout were fed for 10 weeks with either a carbohydrate-free diet (C-free) or with four experimental diets containing various levels (20 or 40%) and sources of starch (extruded wheat or peas) in order to examine metabolic utilisation of dietary vegetable carbohydrates and its endocrine control. The study was focused on the parameters described as limiting in glucose metabolism in fish. Feeding trials were conducted at 8 and 18 °C to establish whether carbohydrate-rich diets can be used in trout farming irrespective of water temperature. At both temperatures, pea diets (especially the highest level) resulted in a feed efficiency as high as the C-free diet. Fish had similar growth rates except when fed the low wheat content diet. Glycaemia values 6 h after feeding were significantly higher in trout fed carbohydrate diets than those given the C-free diet, whereas plasma insulin levels were similar independently of the levels of dietary starch. This study provides the first evidence that glucokinase (GK) activity and mRNA level in trout liver increase in proportion to the content of dietary starch. Nevertheless, these changes were not correlated with plasma insulin levels. Insulin-like growth factor-I (IGF-I) binding and number of receptors in skeletal muscle were consistently higher than those for insulin but no diet-induced differences were found for any of these parameters. Temperature clearly affected the postprandial profile of glucose and insulin, which both showed lower levels 6 h after feeding at 8 °C than at 18 °C, which was consistent with a lower feed intake. Glucose and insulin levels decreased markedly 24 h after feeding at 18 °C, while they were still high at 8 °C, an observation concordant with delayed transit rate. These findings indicate satisfactory adaptation of rainbow trout to diets with a relatively high vegetable starch content, especially when provided as extruded peas, and indicate that diets with increased levels of carbohydrates can be used in this species even when it is reared at low temperature. [Copyright &y& Elsevier]

Abstract: The distribution of lipids in the flesh of large size brown trout (mean body weight 3.8±0.2 kg) was analyzed using magnetic resonance imaging (MRI) scanning. Assays were conducted with fish from a control line and from a line selected for growth rate, both fed a low- or a high-energy diet for 4 months. The MRI acquisition sequence used was 15 ms echo time (TE), 140 ms repetition time (TR) and 90° flip angle. Based on preliminary complete scanning of one fish, 15 different sections were defined in three areas designated by external anatomical references. The images were corrected for different scanning biases, and the voxel units were calibrated for lipid content of the flesh using external references. Values obtained through MRI matched well with those obtained by chemical and NMR analyses of lipid content (r2>0.91). Automated image analysis was conducted to calculate lipid content in the different muscle regions. Wide differences were observed, demonstrating both anteroposterior and central to periphery gradients. These differences are discussed in relation to tissue distribution, muscle activity and energy requirement for locomotion. There was no difference in lipid content or localization between the two lines. Feeding a high-energy diet resulted in significantly higher lipid deposition, especially in the anterior region of the muscle. [Copyright &y& Elsevier]

Two diets based on fish meal (FM) or on a mixture of plant protein sources supplemented with free amino acids (PP) were fed to rainbow trout having a mean initial body weight of 162.5 g reared at 17±1 °C for 24 weeks to evaluate the effect of plant protein sources on fish growth, morphological and body quality traits. At the end of the growth study, samples were withdrawn for chemical analyses as well as measurements of different parameters of quality.Compared to the FM group, the trout fed diet PP had lower growth rates (mean final body weight 663 vs. 800 g), feed efficiency (0.95 vs. 1.07) and protein efficiency ratio (1.96 vs. 2.13). Trout fed diet PP were also characterised by a lower dressed weight (89.45% vs. 91.07%), hepato-somatic index (0.92% vs. 1.01%) and fillet lipid content (5.50% vs. 6.98%), but had a higher mesenteric fat (5.65% vs. 4.08%), viscero-somatic index (10.55% vs. 8.93%) and fillet crude protein content (20.53% vs. 19.93%). The muscle fatty acid composition reflected that of the diet with a higher level of polyunsaturated (n-6) fatty acids in trout fed diet PP and a higher content in polyunsaturated n-3 fatty acids and n-3/n-6 ratio in trout fed diet FM. Sensory analysis was carried out at 5 days post-mortem on microwave-cooked dorsal portions of fillets stored at 1 °C with ice covering. The results of the triangular test showed differences between groups (P<0.01) in organoleptic characteristics as well as in visual colour. Descriptive tests pointed out that fillets of trout fed diet PP exhibited higher hardness, less sweetness and less odour intensity than trout fed diet FM.In summary, data show that long-term feeding a diet in which fish meal was totally replaced by a mixture of plant protein sources significantly affects growth and quality criteria such as morphometric traits, fat deposits, fillet chemical composition and organoleptic characteristics of large commercial size rainbow trout. [Copyright &y& Elsevier]

The long-term effect of a plant (P)-based diet was assessed by proton nuclear magnetic resonance (1H-NMR) metabolomics in rainbow trout fed a marine fish meal (FM)–fish oil (FO) diet (M), a P-based diet and a control commercial-like diet (C) starting with the first feeding. Growth performances were not heavily altered by long-term feeding on the P-based diet. An 1H-NMR metabolomic analysis of the feed revealed significantly different soluble chemical compound profiles between the diets. A set of soluble chemical compounds was found to be specific either to the P-based diet or to the M diet. Pterin, a biomarker of plant feedstuffs, was identified both in the P-based diet and in the plasma of fish fed the P-based diet. 1H-NMR metabolomic analysis on fish plasma and liver and muscle tissues at 6 and 48 h post feeding revealed significantly different profiles between the P-based diet and the M diet, while the C diet showed intermediate results. A higher amino acid content was found in the plasma of fish fed the P-based diet compared with the M diet after 48 h, suggesting either a delayed delivery of the amino acids or a lower amino acid utilisation in the P-based diet. This was associated with an accumulation of essential amino acids and the depletion of glutamine in the muscle, together with an accumulation of choline in the liver. Combined with an anticipated absorption of methionine and lysine supplemented in free form, the present results suggest an imbalanced essential amino acid supply for protein metabolism in the muscle and for specific functions of the liver. [ABSTRACT FROM AUTHOR]

To meet the growing demand of fish feed for aquaculture, an increasing proportion of marine ingredients are being replaced by blends of plant products. However, the total replacement of marine ingredients in salmonid diets impairs fish performance. This is particularly true during the early fry stage and this stage is therefore considered of particular importance. In rainbow trout (RBT), the existence of a genetic variability to survive and grow with plant-based diets devoid of marine ingredients has now been proved, but the mechanisms behind are little studied especially at early stage. To investigate these, we analysed the whole transcriptome of three isogenic lines of RBT fry, which have similar growth when fed a marine resources-based diet (M diet) but which highly differ in their responses to a plant-based diet (V diet). Analysis of transcriptomes profiles revealed 1740, 1834 and 246 probes differentially expressed among the three genotypes when fed the V diet. The use of these lines led to the discovery of potential molecular markers linked to plant-based diet utilisation, some of them belonging to new pathways, never described before. An important number of genes was related to immunity, but further investigations are needed to better understand the difference between the genotypes in their immune status response to V diet exposure. Finally, differences in expression of genes related to feed intake and sensory perception among genotypes suggested that the mechanisms underlying the differences in growth on plant-based diet are closely linked to diet acceptance. Research on plants components affecting feed intake should be thus further explored. [ABSTRACT FROM AUTHOR]

Terrestrial plant resources are increasingly used as substitutes for fish meal and fish oil in fish feed in order to reduce the reliance of aquaculture on marine fishery resources. Although many studies have been conducted to assess the effects of such nutritional transition, no whole breeding cycles of fish fed diets free from marine resources has been reported to date. We therefore studied the reproductive performance of trout after a complete cycle of breeding while consuming a diet totally devoid of marine ingredients and thus of n-3 long chain polyunsaturated fatty acids (n-3 LC-PUFAs) that play a major role in the formation of ova. Two groups of female rainbow trout were fed from first feeding either a commercial diet (C, marine and plant ingredients), or a 100% plant-based diet (V, blend of plant proteins and vegetable oils). Livers, viscera, carcasses and ova were sampled at spawning and analyzed for lipids and fatty acids. Although the V-diet was devoid of n-3 LC-PUFAs, significant amounts of EPA and DHA were found in livers and ova, demonstrating efficient bioconversion of linolenic acid and selective orientation towards the ova. Some ova were fertilized to assess the reproductive performance and offspring survival. We observed for the first time that trout fed a 100% plant-based diet over a 3-year breeding cycle were able to produce ova and viable alevins, although the ova were smaller. The survival of offspring from V-fed females was lower (-22%) at first spawning, but not at the second. Our study showed that, in addition to being able to grow on a plant-based diet, rainbow trout reared entirely on such a diet can successfully produce ova in which neo-synthesized n-3 LC-PUFAs are accumulated, leading to viable offspring. However, further adjustment of the feed formula is still needed to optimize reproductive performance. [ABSTRACT FROM AUTHOR]

Two experimental rainbow trout lines were developed through divergent selection for low (Lean ‘L’ line) or high (Fat ‘F’ line) muscle fat content. Previous nutritional studies suggested that these lines differed in their regulation of lipid and glucose metabolism. Since insulin acts as an anabolic hormone by regulating lipid and glucose metabolism, we put forward the hypothesis that F line might have a stronger sensitivity to insulin than L line. In order to test this hypothesis, bovine insulin was injected into rainbow trout of the two lines fasted for 48 h. As expected, insulin induced hypoglycemia and activated Akt-TOR signaling both in the liver and muscle of the two lines. We demonstrate that this was coupled with increased expression of insulin dependent glucose transporter (GLUT4) and transcription factors of fatty acid anabolism (LXR and SREBP1c) in the muscle and liver, respectively, and lower mRNA levels of fatty acid oxidation enzymes (CPT1a, CPT1b and HOAD) in the white muscle of both lines. Regarding the genotype effect, TOR signaling response to insulin was stronger in F line as reflected by the higher phosphorylation of S6 protein and elevated mRNA levels of lipogenic enzyme (FAS) in the liver of F line. This observation was concordant with the higher plasma concentrations of free fatty acids and triglycerides in F line. Moreover, mRNA levels of hepatic gluconeogenic enzymes (G6Pase2, FBPase and PEPCK) and muscle fatty acid oxidation enzymes (CPT1a, CPT1b, HOAD and ACO) were higher in the F line. However, very few insulin-genotype interactions were detected, indicating that insulin induced similar changes in lipid and glucose metabolism in both lines. [ABSTRACT FROM AUTHOR]

High levels of dietary lipids are incorporated in feeds for most teleost fish to promote growth and reduce nitrogen waste. However, in Senegalese sole (Solea senegalensis) previous studies revealed that increasing the level of dietary lipids above 8% negatively affect growth and nutrient utilization regardless of dietary protein content. It has been shown that glucose regulation and metabolism can be impaired by high dietary fat intake in mammals, but information in teleost fish is scarce. The aim of this study was to assess the possible effect of dietary lipids on glucose metabolism in Senegalese sole with special emphasis on the regulation of proteins involved in the muscle insulin-signalling pathway. Senegalese sole juveniles (29 g) were fed two isonitrogenous diets (53% dry matter) for 88 days. These two diets were one with a high lipid level (∼17%, HL) and a moderate starch content (∼14%, LC), and the other being devoid of fish oil (4% lipid, LL) and with high starch content (∼23%, HC). Surprisingly, feeding Senegalese sole the HL/LC diet resulted in prolonged hyperglycaemia, while fish fed on LL/HC diet restored basal glycaemia 2 h after feeding. The hyperglycaemic phenotype was associated with greater glucose-6-phosphatase activity (a key enzyme of hepatic glucose production) and lower citrate synthase activity in the liver, with significantly higher liver glycogen content. Sole fed on HL/LC diet also had significantly lower hexokinase activity in muscle, although hexokinase activity was low with both dietary treatments. The HL/LC diet was associated with significant reductions in muscle AKT, p70 ribosomal S6-K1 Kinase (S6K-1) and ribosomal protein S6 (S6) 2 h after feeding, suggesting down regulation of the AKT-mTOR nutrient signalling pathway in these fish. The results of this study show for the first time that high level of dietary lipids strongly affects glucose metabolism in Senegalese sole. [ABSTRACT FROM AUTHOR]

Abstract: A growth trial was conducted to test the growth potential and nutrient utilization of Senegalese sole fed diets with increasing substitution of supplemental fish oil (FO) by vegetable oil (VO) blends. Triplicate groups of twenty Senegalese sole juveniles (12g) were fed to satiation over a period of 12weeks with 6 extruded diets containing 570g protein/kg DM and 90g lipid/kg DM. Two blends of VO were tested (A and B) with two FO substitution rates 50% (VO50A and VO50B) and 100% (VO100A and VO100B). A concomitant replacement of 50% fish meal and 50% FO (VO50PP), and a control diet (CTR) containing only FO, were also evaluated. After 12-weeks feeding the dietary treatments did not affect growth performance and final body composition. Muscle eicosapentaenoic acid (EPA) was reduced in all treatments compared to CTR, but docosahexaenoic acid (DHA) was only reduced in the VO50PP group. FO substitution led to a general increase of muscle linoleic acid (18:2 n−6, LOA) with VO50PP inducing maximal levels (15% vs 6% in FO diet). Lipogenic enzymes (FAS, ME and G6PD) as well as long chain fatty acid elongation (elov5) and desaturation (Δ4 desaturase) were not affected by dietary treatments. Results suggest that Senegalese sole can cope with high levels of VO without compromising growth performance or nutrient utilization. Despite differences in muscle fatty acid profile, fish fillet had good nutritional value. [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]

Abstract: Two lines of rainbow trout divergently selected for muscle fat content, fat line (F) and lean line (L) were used to investigate the effect of genetic selection on digestion, intestinal nutrient transport and fatty acid bioconversion, in relation to dietary starch intake. This study involved a digestibility trial for 2weeks using Cr2O3 as inert marker, followed by a feeding trial for 4weeks. For the entire duration, juvenile trout from the two lines were fed diets with or without gelatinized starch. Blood, pyloric ceca, midgut and hindgut were sampled at 24h after the last meal. Transcripts of the proteins involved in nutrient transport and fatty acid bioconversion were abundant in the proximal intestine. GLUT2 transcripts were slightly higher in the F line ceca than in the L line. Dietary starch intake did not enhance the transcription of intestinal glucose transporters, SGLT1 and GLUT2; but it was associated with the higher expression of ApoA1 and PepT1 in the midgut. Significantly, the F line exhibited higher intestinal mRNA levels of MTP, ApoA4, Elovl2, Elovl5 and D6D than the L line, linked to chylomicron assembly and fatty acid bioconversion. Apparent digestibility coefficients of protein, lipid and starch were high in both lines, but not significantly different between them. In conclusion, we found a higher potential of chylomicron synthesis and fatty acid bioconversion in the intestine of F line, but no adaptive transcriptional response of glucose transporters to dietary starch and no genotypic differences in nutrient digestibility. [Copyright &y& Elsevier]

Genetic adaptation to dietary environments is a key process in the evolution of natural populations and is of great interest in animal breeding. In fish farming, the use of fish meal and fish oil has been widely challenged, leading to the rapidly increasing use of plant-based products in feed. However, high substitution rates impair fish health and growth in carnivorous species. We demonstrated that survival rate, mean body weight and biomass can be improved in rainbow trout (Oncorhynchus mykiss) after a single generation of selection for the ability to adapt to a totally plant-based diet (15.1%, 35.3% and 54.4%, respectively). Individual variability in the ability to adapt to major diet changes can be effectively used to promote fish welfare and a more sustainable aquaculture. [ABSTRACT FROM AUTHOR]

Abstract: In the last years, the increase of aquaculture production has led to the evolution of feed composition with an increasing substitution of fish meal and fish oil with terrestrial plant products. In the meantime, selective breeding of fish has been widely developed. The ability to grow on plant-based diets has recently been proven to be genetically variable, pointing out the interest to increase knowledge on the potential consequences of substitution of fish meal and fish oil on current breeding programs. Moreover, heritabilities of major production traits other than growth also need to be estimated in this new environment. Experimental rainbow trouts (about 3000) were derived from a factorial mating design (25 sires and 10 dams), separated in two batches and fed two experimental diets since the first meal. The marine diet (M) was based on fish meal (FM) and fish oil (FO), while the substitution rate by plant products was total in the plant-based (PB) diet (free of FM and FO). We estimated heritabilities under both diets and genotype by diet interactions for body weight, fork length, condition factor, TGC (thermal-unit growth coefficient) and processing yields: viscera yield, carcass yield, filet yield and head yield. Due to the reduced growth of fish fed the plant-based diet, the estimation of genetic parameters was done on fish from both dietary treatments measured at the same age and also on fish measured at the same weight assuming that family rankings could evolve with further growth. Survival rates were high and did not depend on the diet. At 343days, the body weight of the M batch (440.2g±11.3) was twice that of the PB batch (216.0g±12.6). After 465days, the PB batch reached 407.4g±17.1. Whatever the comparison (at same age or same weight), there were significant diet-induced differences for processing traits except for head yield (P =0.80, same age) and filet yield (P =0.82, same weight). The main findings are the high heritabilities observed in fish fed the plant-based diet for body weight (0.69±0.13), TGC (0.65±0.12) and the generally low, though significant, genotype by diet interactions for final body weight, fork length, carcass yield, viscera yield, head yield and filet yield between fish fed source-contrasted diets. Estimates of genetic correlation between diets were the lowest for early weight (0.67±0.12) and indicate differential abilities for initial growth between M and PB diets. [Copyright &y& Elsevier]

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]

Genetic selection is commonly used in farm animals to manage body fat content. In rainbow trout, divergent selection for low or high muscle fat content leads to differences in utilization of dietary energy sources between the fat muscle line (FL) and the lean muscle line (LL). To establish whether genetic selection on muscle fat content affects the hepatic insulin/nutrient signaling pathway, we analyzed this pathway and the expression of several metabolism-related target genes in the livers of the two divergent lines under fasting and then refeeding conditions. Whereas glycemia returned to basal level 24 h after refeeding in FL trout, it remained elevated in the LL trout. Target of rapamycin (TOR) protein was more abundant in the livers of FL trout than in LL trout, and refeeding activation of the hepatic TOR signaling pathway (TOR, S6K1, and S6) was therefore enhanced. Genes related to glycolysis (glucokinase and pyruvate kinase) and gluconeogenesis (glucose-6-phosphatase and phosphoenolpyruvate carboxykinase) were only slightly affected by refeeding and genetic selection. Refeeding stimulated expression of lipogenic genes and the sterolresponsive element binding protein (SREBP1), and expression of fatty acid synthase, glucose-6-phosphate dehydrogenase, and serine dehydratase was predominant in the livers of FL fish compared with LL fish. In agreement with recent findings linking TOR to lipogenesis control, we concluded that genetic selection for muscle fat content resulted in overactivation of the TOR signaling pathway-associated lipogenesis and probably also improved utilization of glucose. [ABSTRACT FROM AUTHOR]

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]

Abstract: The objective of this study was to characterize the body composition, nutrient utilization, energy storage sites and major economic traits of trout selected at pan-size for or against body weight corrected muscle fat content (fat line, FL and lean line, LL respectively). The study focused on the effect of selection in fish size larger than the size where selection has been applied, and the possible effect of diet composition on the differences between lines. FL and LL trout were fed two diets differing for energy content and protein/lipid ratio during 62-days. The feeding trial (day 370 to day 432) started with 327g fish. Dietary protein and lipid contents were 58.7% and 8.3% dry matter (DM) respectively for diet D8 and 47.7% and 26.7% DM for diet D27. Growth, feed intake, feed utilization and traits related to body shape and composition were recorded. In both lines diet D27 improved growth rate, feed efficiency, protein retention and fat gain (P <0.05), and was associated with a higher viscero-somatic index, fillet fat content and lower trimmed fillet yield than diet D8. At the end of the trial, LL fish were slightly heavier than FL ones (P =0.02, no line×diet interaction). The main differences between lines were observed in lipid gain and retention. Values of lipid retention (% of intake) were higher in FL fish (P =0.04) regardless of diet, and suggested a greater ability of FL fish in de novo lipid synthesis. Fat deposition (% of weight gain) was greater in FL fish during the trial and occurred in fillet (P =0.04), on internal side of the belly flap (P =0.04), as well as in other sites of the body, as indicated by higher overall body lipid content at day 432 (P =0.02, no line×diet interaction). Nevertheless, carcass and fillet yields were not different between lines regardless of diet. In conclusion, selection for body weight corrected muscle fat content modified the ability of the fish to utilize nutrients and to store more or less fat in the different body sites. The differences were expressed in a large range of dietary protein/fat ratios. Line×diet interactions were recorded for a very limited number of traits, indicating that the combined use of genetic and nutritional tools should be efficient to manage carcass quality (growth, body shape, processing yields) and fat deposition (quantity and body location) in trout, with no need to adapt diet formulation according to lines. [Copyright &y& Elsevier]

Abstract: In mammals, carnitine palmitoyltransferase 1 (CPT1) is a rate limiting enzyme of fatty acid oxidation. Two isoforms are present. We characterized a full-length cDNA sequence encoding chicken liver L-CPT1 isoform and a partial cDNA sequence encoding chicken muscle M-CPT1 isoform. CPT1 messengers showed the expected tissue specificity. M-CPT1 messenger and CPT1 activity were higher in oxidative than in glycolytic muscle. Expression of both isoforms was assessed in various tissues of genetically fat or lean chickens. Fasting considerably increased L-CPT1 mRNA expression and β-hydroxyacyl CoA dehydrogenase (HAD) activity in the liver of fat or lean chickens. Unexpectedly, fasting did not increase M-CPT1 mRNA levels nor HAD activity in muscles of either chicken genotype. It however increased succinyl-CoA:3-ketoacid CoA transferase (SCOT) mRNA expression (an enzyme related to ketone body utilization) in oxidative muscle. SCOT messenger was slightly more abundant in oxidative muscle of lean chickens but not in glycolytic muscle. In conclusion, the regulation of fatty acid oxidation is probably not impaired in fat chicken. The absence of fasting stimulation of M-CPT1 mRNA expression, which is at variance with the situation observed in mammals, suggests that during fasting, chicken muscles preferentially use ketone bodies as fuel, at least in the short term. [Copyright &y& Elsevier]

The role of somatolactin (SL) in the regulation of energy homeostasis in gilthead sea bream (Sparus aurata) has been analysed. First, a down-regulation of plasma SL levels in response to gross shifts in dietary amino acid profile and the graded replacement of fish meal by plant protein sources (50%, 75% and 100%) has been observed. Thus, the impaired growth performance with changes in dietary amino acid profile and dietary protein source was accompanied by a decrease in plasma SL levels, which also decreased over the course of the post-prandial period irrespective of dietary nitrogen source. Secondly, we examined the effect of SL and growth hormone (GH) administration on voluntary feed intake. A single intraperitoneal injection of recombinant gilthead sea bream SL (0.1 μg/g fish) evoked a short-term inhibition of feed intake, whereas the same dose of GH exerted a marked enhancement of feed intake that still persisted 1 week later. Further, we addressed the effect of arginine (Arg) injection upon SL and related metabolic hormones (GH, insulin-like growth factor-I (IGF-I), insulin and glucagon) in fish fed diets with different nitrogen sources. A consistent effect of Arg injection (6.6 μmol/g fish) on plasma GH and IGF-I levels was not found regardless of dietary treatment. In contrast, the insulinotropic effect of Arg was found irrespective of dietary treatment, although the up-regulation of plasma glucagon and glucose levels was more persistent in fish fed a fish meal based diet (diet FM) than in those fed a plant protein diet with a 75% replacement (diet PP75). In the same way, a persistent and two-fold increase in plasma SL levels was observed in fish fed diet FM, whereas no effect was found in fish fed diet PP75. Taken together, these findings provide additional evidence for a role of SL as a marker of energy status, which may be perceived by fish as a daily and seasonal signal of abundant energy at a precise calendar time. [Copyright &y& Elsevier]

The carnitine palmitoyltransferase I (EC.2.3.1.21; CPT I) mediates the transport of fatty acids across the outer mitochondrial membrane. In mammals, there are two different proteins CPT I in the skeletal muscle (M) and liver (L) encoded by two genes. The carnitine palmitoyltransferase system of lower vertebrates received little attention. With the aim of improving knowledge on the CPT family in fish, we examined CPT I cDNA and CPT activity in different tissues of rainbow trout (Oncorhynchus mykiss). Using RT-PCR, we successfully cloned a partial CPT I cDNA sequence (1650 bp). The predicted protein sequence revealed identities of 63% and 61% with human L-CPT I and M-CPT I, respectively. This mRNA is expressed in liver, white and red skeletal muscles, heart, intestine, kidney and adipose tissue of trout. This is in good agreement with the measurement of the CPT activity in the same tissues. The [IC50] that reflects the sensitivity to malonyl-CoA inhibition was 0.116±0.004 μM for the liver and 0.426±0.041 μM for the white muscle. These results demonstrate for the first time the existence of at least one gene encoding for CPT I present in both the liver and the muscle of rainbow trout. [Copyright &y& Elsevier]