Inge Geurden

Rainbow trout can discriminate between feeds with different oil sources

The purpose of present two-choice trials was to examine the capacity of groups of juvenile rainbow trout to differentiate between two isolipidic diets containing distinct oils and to detect an eventual preference. The choice was offered by means of two self-feeders per tank. One feeder distributed a standard diet with fish oil (FO), the other a diet containing vegetable oil, either rich in linolenic acid (linseed oil, LO), linoleic acid (sunflower oil, SO), or oleic acid (rapeseed oil, RO). Each 15-day preference test was preceded by a 15-day adaptation period during which both feeders distributed the same diet. The tests were followed by a 10- to 15-day validation period in order to confirm that feeder solicitations were steered by the characteristics of the diets. Preferences were expressed as relative changes in feed demands for a specific feeder. Averaged over all groups, the preference tests demonstrated the capacity of rainbow trout to discriminate between a diet with FO and a diet containing vegetable oil, and indicated a general preference for the diet with FO over the other diets irrespective of whether they received the diet with fish oil (Experiment 1) or with vegetable oil (Experiment 2) prior to the preference test. The tests also indicated a difference in the extent of relative avoidance of each of the three vegetable oil diets. Diet LO was the most avoided, as indicated by the 37-39% decrease in demands for the feeder with diet LO (P<0.05). Diet RO was the best accepted, causing a decrease in feed demands of only 15-17% (P>0.05). The avoidance of diet SO at the end of the preference test was 30% (P>0.05) after an initially higher avoidance of 43% (P<0.05). It is believed that the metabolic consequences of the excess of linolenic or linoleic acid negatively affected the feed acceptances of diets LO and SO. Further work is needed to elucidate a possible interference of differences in palatability. In all groups, the lower demands for the vegetable oil diets were compensated by increased demands for diet FO. Hence, changes in diet selection had no effect on total feed or energy intakes, measured as the sum of both selections.

Hepatic protein kinase B (Akt)-target of rapamycin (TOR)-signalling pathways and intermediary metabolism in rainbow trout (Oncorhynchus mykiss) are not significantly affected by feeding plant-based diets.

The aim of the present study was to analyse the effects of partial or total replacement of fish meal (FM) and fish oil (FO) by a mixture of plant protein (PP) and a mixture of vegetable oils (VO) on the hepatic insulin-nutrient-signalling pathway and intermediary metabolism-related gene expression in rainbow trout (Oncorhynchus mykiss). Triplicate groups of fish were fed four practical diets containing graded levels of replacement of FM and FO by PP and VO for 12 weeks: diet 0/0 (100 % FM, 100 % FO); diet 50/50 (50 % FM and 50 % PP, 50 % FO and 50 % VO); diet 50/100 (50 % FM and 50 % PP, 100 % VO); diet 100/100 (100 % PP, 100 % VO). Samplings were performed on trout starved for 5 d then refed with their allocated diet. In contrast to partial substitution (diet 50/50), total substitution of FM and FO (diet 100/100) led to significantly lower growth compared with diet 0/0. The insulin-nutrient-signalling pathway (protein kinase B (Akt), target of rapamycin (TOR), S6 protein kinase 1 (S6K1) and S6) was characterised in trout liver and found to be activated by refeeding. However, changes in diet compositions did not differentially affect the Akt-TOR-signalling pathway. Moreover, expression of genes encoding fructose-1,6-biphosphatase, mitochondrial phosphoenolpyruvate carboxykinase, glucokinase, pyruvate kinase and carnitine palmitoyl transferase 1 were not affected by refeeding or by dietary changes. Refeeding down- and up-regulated the expression of gluconeogenic glucose-6-phosphatase isoform 1 and lipogenic fatty acid synthase genes, respectively. Expression of both genes was also increased with partial replacement of FM and total replacement of FO (diet 50/100). These findings indicate that plant-based diets barely affect glucose and lipid metabolism in trout.

High levels of dietary fat impair glucose homeostasis in rainbow trout

SUMMARY 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.

High or low dietary carbohydrate:protein ratios during first-feeding affect glucose metabolism and intestinal microbiota in juvenile rainbow trout

Based on the concept of nutritional programming in mammals, we tested whether an acute hyperglucidic–hypoproteic stimulus during first feeding could induce long-term changes in nutrient metabolism in rainbow trout. Trout alevins received during the five first days of exogenous feeding either a hyperglucidic (40% gelatinized starch + 20% glucose) and hypoproteic (20%) diet (VLP diet) or a high-protein (60%) glucose-free diet (HP diet, control). Following a common 105-day period on a commercial diet, both groups were then challenged (65 days) with a carbohydrate-rich diet (28%). Short- and long-term effects of the early stimuli were evaluated in terms of metabolic marker gene expressions and intestinal microbiota as initial gut colonisation is essential for regulating the development of the digestive system. In whole alevins (short term), diet VLP relative to HP rapidly increased gene expressions of glycolytic enzymes, while those involved in gluconeogenesis and amino acid catabolism decreased. However, none of these genes showed persistent molecular adaptation in the liver of challenged juveniles (long term). By contrast, muscle of challenged juveniles subjected previously to the VLP stimulus displayed downregulated expression of markers of glycolysis and glucose transport (not seen in the short term). These fish also had higher plasma glucose (9 h postprandial), suggesting impaired glucose homeostasis induced by the early stimulus. The early stimulus did not modify the expression of the analysed metabolism-related microRNAs, but had short- and long-term effects on intestinal fungi (not bacteria) profiles. In summary, our data show that a short hyperglucidic–hypoproteic stimulus during early life may have a long-term influence on muscle glucose metabolism and intestinal microbiota in trout.

Link between lipid metabolism and voluntary food intake in rainbow trout fed coconut oil rich in medium-chain TAG.

We examined the long-term effect of feeding coconut oil (CO; rich in lauric acid, C12) on voluntary food intake and nutrient utilisation in rainbow trout (Oncorhynchus mykiss), with particular attention to the metabolic use (storage or oxidation) of ingested medium-chain TAG. Trout were fed for 15 weeks one of the four isoproteic diets containing fish oil (FO) or CO as fat source (FS), incorporated at 5% (low fat, LF) or 15% (high fat, HF). Fat level or FS did not modify food intake (g/kg(0·8) per d), despite higher intestinal cholecystokinin-T mRNA in trout fed the HF-FO diet. The HF diets relative to the LF ones induced higher growth and adiposity, whereas the replacements of FO by CO resulted in similar growth and adiposity. This, together with the substantial retention of C12 (57% of intake), suggests the relatively low oxidation of ingested C12. The down-regulation of carnitine palmitoyl-transferase-1 (CPT-1) confirms the minor dependency of medium-chain fatty acids (MCFA) on CPT-1 to enter the mitochondria. However, MCFA did not up-regulate mitochondrial oxidation evaluated using hepatic hydroxyacyl-CoA dehydrogenase as a marker, in line with their high retention in body lipids. At a low lipid level, MCFA increased mRNA levels of fatty acid synthase, elongase and stearoyl-CoA desaturase in liver, showing the hepatic activation of fatty acid synthesis pathways by MCFA, reflected by increased 16 : 0, 18 : 0, 16 : 1, 18 : 1 body levels. The high capacity of trout to incorporate and transform C12, rather than to readily oxidise C12, contrasts with data in mammals and may explain the absence of a satiating effect of CO in rainbow trout.

The Positive Impact of the Early-Feeding of a Plant-Based Diet on Its Future Acceptance and Utilisation in Rainbow Trout

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.

Macronutrient-induced differences in food intake relate with hepatic oxidative metabolism and hypothalamic regulatory neuropeptides in rainbow trout (Oncorhynchus mykiss).

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

The evaluation of energy intake adjustments and preferences in juvenile rainbow trout fed increasing amounts of lipid.

This study aimed at a better understanding of the feed intake (FI) regulation in rainbow trout by dietary digestible energy (DE). The DE contents of the three diets (20.5, 23.0 and 24.7 kJ per g dry diet) were modified by supplementing different amounts of fish oil. The crude lipid and protein levels were 13%, 26% and 34% and 64%, 54% and 48%, respectively (% dry diet). The daily FI was measured by means of self-feeders in groups of rainbow trout (32-55 g, initial BW) during two 5 to 6-week trials. Their eventual preference for one of the lipid levels was evaluated by offering the choice between a low and higher lipid diet. The results indicated that fish of a similar body mass had a similar FI without apparent energy intake compensations. It is believed that the excessive energy intakes with the higher lipid diets were not caused by a higher palatability of these diets since the trout did not express any particular preference. The trout fed the lipid rich diets had a higher level of body adiposity, but a similar protein growth. The observation that the trout did not reduce FI when fed the high lipid diet implies a low negative feedback by the ingested fat or by the increase in body adiposity. The similarities in lean carcass growth favour the idea that growing animals regulate their FI in order to meet the demand for maximal protein growth rather than to satisfy a predetermined energy requirement.

Constraints on Energy Intake in Fish: The Link between Diet Composition, Energy Metabolism, and Energy Intake in Rainbow Trout

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 (2×2 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 non-protein 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.

Maintenance and growth requirements for nitrogen, lysine and methionine and their utilisation efficiencies in juvenile black tiger shrimp, Penaeus monodon, using a factorial approach

We used a factorial approach to distinguish maintenance from growth requirements for protein, lysine and methionine in the black tiger shrimp, Penaeus monodon. Juvenile P. monodon (initial weight 2.4 g) were fed during 6 weeks one of ten semi-purified diets based on casein and purified amino acids (AA) as N source. The diets contained four levels of crude protein (CP, from 5 to 54 % DM diet) with two levels (% CP) of lysine or methionine (normal or 30 % deficient). Requirements were determined using linear and non-linear regression models. We could thus obtain the first ever data on maintenance (N equilibrium) requirements for CP and AA in P. monodon. CP requirements for maintenance (4.5 g/kg body weight (BW) per d) represented approximately 19 % of the CP requirement for maximal N gain (23.9 g/kg BW per d). The marginal efficiency of utilisation reached a maximum of 38 % for N, 0.77 for lysine and 1.62 for methionine using N gain as response. Lysine requirements were 0.20 g/kg BW per d for N maintenance and 1.40 g/kg BW per d for maximal N gain. Methionine requirements were 0.11 g/kg BW per d for N maintenance and 0.70 g/kg BW per d for maximal N gain. The lysine (5.8 %) and methionine (2.9 %) requirements for maximal N gain, expressed as percentage of protein requirement, agree with literature data using a dose-response technique with smaller P. monodon. The observed interaction between dietary CP and methionine for N gain demonstrates that requirements for indispensable AA (expressed as % CP) cannot be evaluated separately from CP requirements.