Feeding rainbow trout with a lipid-enriched diet: effects on fatty acid sensing, regulation of food intake and cellular signaling pathways.

Using rainbow trout fed with low-fat or high-fat diets, we aimed to determine whether the response of food intake, mRNA abundance of hypothalamic neuropeptides involved in the metabolic regulation of food intake and fatty acid sensing systems in the hypothalamus and liver are similar to results previously observed when levels of specific fatty acids were raised by injection. Moreover, we also aimed to determine if the phosphorylation state of intracellular energy sensor 5'-AMP-activated protein kinase (AMPK), and proteins involved in cellular signaling such as protein kinase B (Akt) and target of rapamycin (mTOR) display changes that could be related to fatty acid sensing and the control of food intake. The increased levels of fatty acids in the hypothalamus and liver of rainbow trout fed with a high-fat diet only partially activated fatty acid sensing systems and did not elicit changes in food intake, suggesting that the fatty acid sensing response in fish is more dependent on the presence of specific fatty acids, such as oleate or octanoate, rather than to the global increase in fatty acids. We also obtained, for the first time in fish, evidence for the presence and function of energy sensors such as AMPK and proteins involved in cellular signaling, like mTOR and Akt, in the hypothalamus. These proteins in the hypothalamus and liver were generally activated in fish fed the high-fat versus low-fat diet, suggesting that cellular signaling pathways are activated in response to the increased availability of fatty acids.

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.

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.

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.

A vegetable oil feeding history affects digestibility and intestinal fatty acid uptake in juvenile rainbow trout Oncorhynchus mykiss.

Future expansion of aquaculture relies on the use of alternatives to fish oil in fish feed. This study examined to what extent the nature of the feed oil affects intestinal lipid uptake properties in rainbow trout. The fish were fed a diet containing fish (FO), rapeseed (RO) or linseed (LO) oil for 8 weeks after which absorptive properties were assessed. Differences in digestibility due to feed oil history were measured using diet FO with an indigestible marker. Intestinal integrity, paracellular permeability, in vitro transepithelial fatty acid transport (3H-18:3n-3 and 14C-16:0) and their incorporation into intestinal epithelia were compared using Ussing chambers. Feed oil history did not affect the triacylglycerol/phosphatidylcholine ratio (TAG/PC) of the newly synthesized lipids in the segments. The lower TAG/PC ratio with 16:0 (2:1) than with 18:3 (10:1) showed the preferential incorporation of 16:0 into polar lipids. The FO-feeding history decreased permeability and increased transepithelial resistance of the intestinal segments. Transepithelial passage rates of 18:3n-3 were higher when pre-fed LO compared to RO or FO. Similarly, pre-feeding LO increased apparent lipid and fatty acid digestibilities compared to RO or FO. These results demonstrate that the absorptive intestinal functions in fish can be altered by the feed oil history and that the effect remains after a return to a standard fish oil diet.

Dietary phosphatidylcholine affects postprandial plasma levels and digestibility of lipid in common carp (Cyprinus carpio).

This study examined the effect of purified soyabean phosphatidylcholine (SPC) on circulating plasma lipids and nutrient digestibility in juvenile common carp. The fish (100 (SD 15) g, 25 degrees C) were fed, for 4 weeks, a casein-based diet containing either 12 % soyabean oil (diet SBO) or 8 % SBO plus 4 % SPC (diet SPC). The lipid, protein and energy contents of the faeces were analysed for the determination of apparent digestibility. At the end of the trial, the fish were fasted for 48 h and fed a single meal. Plasma lipids were then analysed over the next 48 h. The growth (1.63 v. 1.26 % per d) and apparent lipid digestibility (96.3 v. 92.1 %) were higher in SPC- than in SBO-fed fish. The amplitude of the postprandial (8 h after the meal) TAG peak was identical in fish from both treatments, despite the 33 % lower amount of TAG in diet SPC. Both observations support the idea that SPC stimulates intestinal TAG uptakes. The lower TAG:phospholipid ratio of the secreted plasma lipids at the time of absorption suggests a larger number of smaller intestinal lipoproteins in SPC- than SBO-fed fish, possibly due to the recycling of absorbed lysophosphatidylcholine for chylomicron formation. In the 48 h unfed state, phospholipid levels remained approximately 20 % higher in SPC-fed than in SBO-fed fish, but we observed no hypocholesterolaemic effect of SPC. In summary, the present data support earlier histological indications of a positive role of dietary phosphatidylcholine in intestinal TAG uptakes in carp.