The current experiment aimed to study whether interactions with lipid metabolism possibly might explain the relative increased liver weight obtained in fish fed sub-optimal methionine levels. A basal diet based on a blend of plant proteins which is low in methionine (1.6 g Met/16 g N) was compared to a methionine adequate diet (2.2 g Met/16 g N) prepared by adding DL-methionine (2.4 g/kg) to the basal diet in the expense of wheat grain. Fish oil was used as the lipid source. The diets were balanced in all nutrients except methionine. The diets were fed to Atlantic salmon (500 g BW) for a period of 3 months. Feed intake did not differ, rendering the intake of all nutrients except methionine equal. Fish fed the low methionine diet had an increased liver size relative to body weight, indicating fat deposition in the liver. Fish given the sub-optimal methionine diet showed about six times higher fatty acid synthase (FAS) activity as compared to the fish fed the adequate methionine diet, indicating a higher de novo lipogenesis. A significant rise in the liver 18:1 to 18:0 fatty acid ratios also supported storage of lipids over fatty acid oxidation. Indeed, methionine limitation resulted in significantly higher TAG concentrations in the liver. Sub-optimal dietary methionine also resulted in lower hepatic taurine concentrations and the total bile acids concentrations were reduced in faeces and tended to be reduced in plasma. Taken together, our data show that salmon fed sub-optimal methionine levels had increased relative liver weight and developed signs commonly described in the early stage of non-alcoholic fatty liver disease in rodent models (increased FAS activity, changed fatty acid ratios and TAG accumulation).
The appetite-stimulating effect of krill hydrolysate (KH), free amino acids and specific water-soluble low molecular N-compounds was explored in a short feeding trial with Atlantic salmon. A 100 g kg À1 fish meal positive (MFM), a 30 g kg À1 fish meal negative control (LFM) and six more LFM diets were produced added known attractants for fish-KH at two levels: low (LAK) or high (HAK), AMP, choline chloride (CC); an amino acid mix resembling the free amino acid composition of KH (AA) or a mix of AMP, CC and AA (ACA). HAK, ACA and AA showed highest feed intake and significantly higher plasma phospholipids and cholesterol. The lower performing treatments (LFM, AMP) showed higher liver lipids and hepatosomatic index. CC induced reduced liver lipids and increased plasma phospholipids and cholesterol. Appetite regulating neuropeptide gene expression analysis (qPCR) was performed in fish fed LFM, HAK and AA. Pyy showed the highest postprandial expression in LFM, whereas the expression of the anorexigenic neuropeptides cart, pomca1, pomca2 and pomca2s was low. These apparently contradictory results may be explained by initial appetite stimulation by HAK and AA, resulting in higher feed intake, followed by satiation and appetite downregulation.
This experiment aimed to test the interaction of lysine limitation with nutrient accretion and muscle carnitine depot in Atlantic salmon. Fish were fed adequate or low‐lysine diets for 3 months. Lysine intake was significantly less (48%) in fish fed the low‐lysine diet as compared with that fed the adequate one. There was no difference in dietary amino acids between treatments, with the exception of lysine. The lower lysine intake was reflected in plasma free lysine being 52% less while the free lysine concentration in the liver and muscle were unaffected.
Although there was no significant difference between voluntary feed intakes among treatments, fish fed the low‐lysine diet had reduced growth, protein and energy deposition as compared with fish fed the adequate lysine diet. White trunk muscle contained more glycogen and less protein in fish fed the low‐lysine diet while no difference in lipid was observed. The livers from fish fed the low‐lysine diet contained less glycogen and slightly more fat and protein than the livers from fish fed the adequate lysine diet. Lysine limitation reduced carnitine in the liver without affecting muscle carnitine depot. Thus, low‐lysine diets did not likely affect the fatty acid oxidation capacity. This fact was supported by unaffected fatty acid profiles and lipid classes between treatments during the 3‐month study. In conclusion, lysine limitation does not deplete the muscle carnitine depot during the on‐growing seawater phase of Atlantic salmon, but affects the deposition pattern of nutrients.
Digestive enzymes of freshwater fish Labeo rohita (family Cyprinidae; class Actinopterygii; infraclass Teleostei; order Cypriniformes) were studied during ontogenic development. Amylase, protease and lipase activities showed a polynomial relationship with age of rohu, whereas trypsin, chymotrypsin and lipase activities exhibited exponential trends with the age of rohu larvae. SDS‐PAGE of crude enzyme extract revealed that the proteases of higher molecular weight (MW) appeared during early ontogeny, whereas low MW proteases were observed at a later stage. Substrate SDS‐PAGE supported the quantitative study of protease activities as evidenced with the increasing number and intensity of activity bands with the age of fish. The number of protease activity bands observed in 4, 10, 12 and 24 DAH (days after hatching) larvae were 5, 7, 8 and 9, respectively. Inhibition of protease activities with soybean trypsin inhibitor (SBTI) (58.6–81.2%), phenyl methyl sulphonyl fluoride (PMSF) (55.6–70%), N‐α‐p‐tosyl‐l‐lysine chloromethyl ketone (TLCK) (41.1–52.3%) and N‐tosyl‐l‐phenylalanine chloromethyl ketone (TPCK) (27.9–44.5%) indicated the presence of serine proteases, trypsin‐ and chymotrypsin‐like enzymes in rohu larvae. Inhibition with SBTI and TPCK showed power, TLCK and ethylenediaminetetraacetic acid showed exponential, whereas PMSF showed polynomial relationships during the study period.
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