The patterns of plasma free amino acids after force-feeding in rainbow trout Oncorhynchus mykiss (Walbaum) with and without dorsal aorta cannulation

Two experiments were conducted to compare the patterns of plasma free amino acid concentrations after force-feeding in rainbow trout Oncorhynchus mykiss (Walbaum) with and without dorsal aorta cannulation. In the first experiment, 35 rainbow trout averaging 504 ± 7.8 g (mean ± SD) were divided into seven groups of five fish each. After 48 h starvation, a group of fish was anaesthetized and blood samples were taken at one of the following time periods: 0, 4, 8, 12, 24, 36 and 48 h after feeding. In the second experiment, five dorsal aorta cannulated rainbow trout averaging 511 ± 6.2 g (mean ± SD) were kept in a cage. After 48 h starvation, the fish were anaesthetized and blood samples were taken from the same fish at 0, 4, 8, 12, 24, 36 and 48 h after feeding. In the first experiment, the concentration of all plasma free amino acids except histidine and glycine peaked at 4 h and returned to the basal level 24 h after feeding. In the second experiment, the concentration of all plasma free amino acids except isoleucine, leucine, phenylalanine and tryptophan also peaked at 4 h and returned to the basal level 24 h after feeding. These results showed that the pattern of plasma free amino acid concentrations from fish with and without dorsal aorta cannulation were similar.     

Dietary essential amino acids and heat increment in rainbow trout (Oncorhynchus mykiss)

Oxygen consumption attributable to apparent heat increment (AHI) was measured in relation to varying essential amino acid proportions (EAA) infused into rainbow trout,Oncorhynchus mykiss (250–450 g), induced to swim at 1 BL s^–1. Five diets, mimicking EAA concentrations in trout whole body protein, deficient in the branched chain amino acids (isoleucine, leucine and valine), containing unbalanced proportions of EAAs and supplying lysine in excessive and limiting proportions, were tested. Following infusion of the experimental diets, a significant increase in oxygen consumption was observed. Changes in plasma EAAs following infusion paralleled the time course of AHI (i.e., oxygen consumption). AHI represented the equivalent of 15–32% of the gross energy intake depending on dietary EAA composition. Diets supplying EAAs similar to trout whole body protein and limiting in lysine produced the lowest AHI values, indicating efficient utilization of dietary amino acids. Higher AHI values were associated with diets deficient in the branched chain amino acids and diets supplying lysine in excess. Duration of elevated metabolism was independent of both dietary composition and energy intake. Different proportions of EAAs in the diet can increase the energy expended as AHI. In an attempt to reduce the energy liberated as AHI, attention must be paid to the quality, quantity and balance of dietary EAAs.     

Arginine effects on biochemical composition of sperm in rainbow trout,Oncorhynchus mykiss

The present study investigated the effect of arginine on seminal plasma composition in rainbow trout. Male rainbow trout broodstocks (2500 ± 200 g) were fed five practical diets (each consisting of three triplicates) supplemented with Arginine at 0.50%, 1.50% and 2.00%. The control group were fed without arginine. Broodstock feeding lasted for 90 days, and then fish semen was sampled. Results indicated no significant differences in LDH, ALP, Fe²⁺ and phosphorous content among the different treatments. The lowest levels of AST and ALT and the highest levels of Ca²⁺ and Mg²⁺ ions were observed in the treatment fed with 1.50% arginine, which showed significant differences from other treatments (P < 0.05). Moreover, the amount of Cl^?, Na⁺ and K⁺ ions was significantly increased in the seminal plasma in fish fed diets containing arginine in comparison with the control. As the amount of arginine was increased, the levels of uric acid became significantly greater in contrast to urea and glucose levels. The highest amounts of cholesterol, fructose and total protein were observed in treatments fed on 2.00%, 0.50% and 1.00% arginine, respectively, showing significant differences from other treatments (P < 0.05). The highest pH value was assayed in the 1.50% arginine treatment. Results indicated that arginine had a potential efficacy on semen quality in rainbow trout broodstocks.     

Effect of feeding on the tissue free amino acid concentrations in rainbow trout (Oncorhynchus mykiss Walbaum)

This study investigates whether tissue free amino acid (FAA) pools in rainbow trout,Oncorhynchus mykiss (Walbaum), are altered following feeding and the relationships between the amount of food consumed and the FAA pool size. Trout were starved for 7 days to provide baseline data and then refed on day 8. Individual food intake was measured by radiography and the consumption of amino acids (AA) calculated from dietary protein consumption. Total FAA concentrations in the stomach, liver and white muscle were little changed at various times after the meal and this pattern was repeated for the majority of individual FAA. Overall, the most notable change was a reduction in essential FAA concentrations (principally in valine, leucine and isoleucine) in the white muscle following feeding. However, in the caeca total FAA, total essential FAA and a number of individual FAA were significantly elevated at 4, 9 and 15h following feeding. There were few significant correlations between dietary amino acid consumption and total tissue FAA and essential FAA concentration in the stomach, caecum and white muscle; correlations were stronger in the liver. In order to explain the relative constancy of total FAA concentrations in the tissues following food intake (which represents over 100% of the total FAA pool) a model is presented that quantifies the AA flux through the free pools and considers the role of protein turnover in regulating FAA pool size.     

Bioavailability of amino acids chelated and glass embedded zinc to rainbow trout, Oncorhynchus mykiss, fingerlings

A 2 × 4 factorial experiment was conducted to determine the bioavailability of zinc (Zn) from amino acids chelated (Zn–Am) and glass embedded Zn (Zn–Gl) as sources for rainbow trout, Oncorhynchus mykiss , fed practical type diets. Two levels of Zn (20 and 40 mg kg⁻¹) were supplemented to the diets using either zinc sulphate (Zn–Sf), zinc methionine (Zn–Mt), Zn–Am or Zn–Gl. Rainbow trout with an average weight of 2 g were fed the experimental diets for 15 weeks. Growth and feed gain ratio (FGR) were not significantly influenced by the dietary Zn content and forms. Alkaline phosphatase (ALP) activity for both levels of Zn–Am was significantly higher than that of Zn–Sf and Zn–Gl at 20 mg supplementation. In another experiment, fish of about 95 g were fed the same experimental diets to determine the absorption of Zn and it was found to be significantly higher from Zn–Am compared with the rest. Retention from Zn–Am at 20 mg was significantly higher than the rest, excluding Zn–Sf. The results suggest that the availability of Zn from Zn–Am might be superior among the sources compared.     

Influence of dietary arginine and glycine content on thyroid function and growth of juvenile rainbow trout, Oncorhynchus mykiss (Walbaum)

Juvenile rainbow trout, held at 12°C on a 12 h light :12 h dark photocycle, were fed a constant ration (1 % of body wt day ^?1) of isonitrogenous and isoenergetic diets that varied in either arginine content (3.6-56.1 g kg^?1 dry matter; experiment 1), or glycine (3.3–118 g kg^?1 dry matter) and alanine (5.0-42.3 g kg^?1 dry matter) content (experiment 2). In experiment 1, the lowest dietary level of arginine depressed growth, feed efficiency, plasma l -thyroxine (T₄) and 3,5,3′-triiodo-l -thyronine (T₃) levels and hepatic T₄ 5’monodeiodinase (5'D) activity responsible for T₄-to-T₃ conversion. Over the dietary range of 7.1–56.1 g arginine kg^?1, there was no change in 5'D activity, despite an arginine stimulation of growth. The optimum level of arginine for growth was within the range of 14.1–28.1 g kg^?1 of the diet or 32–63 g kg^?1 of dietary protein. In experiment 2, an increase in dietary glycine level, at the expense of glutamic acid, increased 5'D activity without attendant elevation of the plasma T₃ level. The latter finding suggests that glycine also induced a compensatory increase in T₃ degradation rate. This may explain why the glycine-induced increase in 5'D activity was unaccompanied by any changes in growth indices. Alteration of dietary alanine content did not affect growth or thyroid function. We conclude that of the various dietary amino acids tested, only glycine led to a progressive stimulation of hepatic T₄ 5'D activity. However, because glycine likely enhanced T₃ degradation, no increases in plasma T₃ or growth indices were found. Glycine may serve as an advance signal that activates thyroid function immediately preceding or coincident with energy and nutrient (especially protein) intake. This, in turn, may improve the efficiency of nutrient absorption and/or post-absorptive anabolic events.     

Intraperitoneal injection of sulfur amino acids enhance the hepatic cysteine dioxygenase activity in rainbow trout (Oncorhynchus mykiss)

As part of the investigation into cysteine metabolism in fish, sulfur amino acids and their derivatives were injected intraperitoneally to fingerling rainbow trout (Oncorhynchus mykiss) to examine how the doses of these compounds affect the hepatic cysteine dioxygenase [EC 1.12.11.20] in this species. A dose of 0.25 mmol L-cysteine per 100 g body weight induced the enzyme activity as much as 2.5 times that of the control fish within 4h after the injection. The activity increased proportionally to the increasing dose of cysteine up to the dose of 0.15 mmol per 100 g body weight. The induction was observed to be rather specific to L-cysteine. These findings suggested that the cysteine sulfinate pathway might play an important role in the metabolism of excess cysteine in rainbow trout. The dosage of L-cysteine larger than 0.50 mmol per 100g body weight led to mortality of the fish. The pathway of cysteine catabolism was considered to function to prevent toxic accumulation of cysteine in rainbow trout, as in the case of mammals.     

Cholesterolaemia and triacylglycerolaemia in farmed rainbow trout,Oncorhynchus mykiss

Physiological (reference) value intervals determined by the lower 2.5% and upper 97.5% quantiles were calculated for blood plasma cholesterol (CHOL) and triacylglycerol (TGL) in farmed rainbow trout, Oncorhynchus mykiss, in raceway culture. The fish were given dry pelleted diets that contained 37–47% crude protein and 7–14% crude fat and were kept at a stocking density of 50 kg m^?3 in tanks provided with running freshwater at an ambient temperature of 3–16°C and dissolved oxygen, 8.4–11.5 mg L^?1. Blood was sampled between September and November at a photoperiod of 9:00–13:00 hours: 11:00–15:00 hours (light:dark). Cholesterol levels were significantly (P = 0.0001) greater in males (4.7–12.1 mmol L^?1, n = 34, mean weight 406 ± 138 g) than immature females (3.2–9.7 mmol L^?1, n = 386, mean weight 416 ± 147 g). Physiological range for TGL in immature females and males was 2.4–14.4 mmol L^?1 (n = 249, mean weight 418 ± 149 g). The distribution and density of the quantiles in the tested reference group were made possible by the use of histograms, which showed normal distributions for CHOL in males and in females, but not for TGL, in which a sinistral asymmetry was found. Correlation and regression analyses indicated significant (P = 0.0000) dependence between the CHOL and total protein (r² = 76.2%), CHOL and Fulton's condition factor (r² = 43.3%) and CHOL and absolute weight of liver (r² = 45.5%). Fluctuation in cholesterolaemia and triacylglycerolaemia, depending on nutrition and the aquaculture method is discussed below.     

Responses to prolonged hypoxia by rainbow trout (Salmo gairdneri) I. Free amino acids and proteins in plasma,liver and white muscle

In order to estimate the mobilization of nitrogen compounds for energetic purposes in trout under hypoxic conditions, commercial-size rainbow trout, acclimated to 15°C, were maintained for 10 weeks at an oxygen level of 5.3 ± 0.5 mg/l (hypoxic group) or 8.4 ± 0.4 mg/I (control group), and the changes in tissue concentrations of free amino acids and proteins studied. In animals subjected to hypoxia, there was a decrease in plasma free amino acids involved in gluconeogenesis, liver alanine and aspartic acid, plasma and liver protein concentrations, and muscle free histidine. These results suggest a trend of rainbow trout metabolic activity towards energy production at the expense of anabolism when oxygen availability in water is limited over a long period of time.     

The effect of water-borne magnesium on the dietary magnesium requirement of the rainbow trout (Oncorhynchus mykiss)

Rainbow trout (Oncorhynchus mykiss) (mean initial weight 0.84 g) were fed diets containing graded levels of magnesium (Mg) (78 to 725µg/g) while being exposed to one of several levels in the rearing water (1.4 to 1000 mg/l). Uptake of Mg from the water, in Mg-deficient fish, was linearly related to the water Mg concentrated. It appears that the fish's Mg requirement can be met from either or both the diet or water. Under the experimental conditions, a water-borne concentration of 46 mg/l was calculated to be sufficient to meet the Mg requirement of the fish fed a Mg-free diet.     

The effects of dietary organic or inorganic selenium in rainbow trout (Oncorhynchus mykiss) under crowding conditions

In the present study, the effects of different sources of selenium (Se; sodium selenite or selenomethionine) supplementation on the growth and serum concentrations of oxidative stress markers [malondialdehyde (MDA), 8‐isoprostane, glutathione peroxidase (GSH‐Px) activity] and muscle Se, MDA and heat shock protein 70 (Hsp70) levels in rainbow trouts were evaluated. The fish (n = 360; 0 + years old) with initial average weight of 20 ± 0.8 g were randomly assigned to 12 treatment groups consisting of 3 replicates of 10 fish each in a 2 × 2 × 3 factorial arrangement of treatments (stocking densities, Se sources, Se levels). The fish were kept at low (25 kg m^?3) or high (100 kg m^?3) stocking densities and fed a basal (control) diet or the basal diet supplemented with either 0.15 or 0.30 mg of Se kg^?1 of diet from two different forms: sodium selenite or selenomethionine. High stocking density decreased weight gain, feed intake and feed conversion ratio (FCR) when basal diet was fed (P = 0.001). A linear increase in feed intake and weight gain and improvement in FCR were found in sodium selenite (P = 0.01)‐ or selenomethionine (P = 0.001)‐supplemented fish reared under crowding conditions. Serum and muscle Se levels and serum GSH‐Px activity increased (P = 0.001) linearly, whereas serum and muscle MDA concentrations and serum 8‐isoprostane decreased linearly as dietary sodium selenite (P = 0.01) or selenomethionine (P = 0.001) supplementation increased. Selenomethionine and sodium selenite supplementation decreased Hsp70 in the muscle of fish reared under crowding conditions (P < 0.05). Supplementation with Se improved growth and antioxidant status of fish and the effects of selenomethionine were relatively greater than sodium selenite in the crowded groups. Results suggest that crowding conditions cause significant detrimental effects in rainbow trout indicated by increased oxidative stress, reduced feed intake and body weight gain. ?t also indicates that dietary Se supplementation offers a feasible way of reducing the losses in performance of rainbow trout reared under crowding conditions. Selenomethionine seems to be more effective than sodium selenite and the higer dose in the present study also seems to be more effective than the lower dose.     

Apparent absorption efficiencies of amino acids in rainbow trout, Oncorhynchus mykiss (Walbaum), fed diets containing bacterial single-cell protein

The aim of the present study was to compare the apparent absorption efficiency of individual indispensable and dispensable amino acids in diets containing varying proportions of bacterial single cell protein (BSCP) and fishmeal when fed to rainbow trout, Oncorhynchus mykiss (Walbaum). Four diets were formulated to contain a total of 45.8% crude protein, of which 0% was from BSCP in diet 1 (BSCP-0), 25% in diet 2 (BSCP-25), 62.5% in diet 3 (BSCP-62.5) and 100% in diet 4 (BSCP-100); the remainder of the protein was from fishmeal. The four diets were isonitrogenous and had similar amino acid compositions. Duplicate groups of 25 fish were fed one of the four experimental diets at the rate of 20 g kg^?1 body weight per day for 132 days. Apparent absorption efficiency was measured over a 45-day period during the growth trial. Results from two methods of faeces collection and two indicators of absorption efficiency were compared. The amino acid absorption efficiency of BSCP-100 was significantly lower (P < 0.05) when compared to the other three diets and absorption of some of indispensable amino acids was below the requirement level. The variation seen in individual amino acids absorption efficiencies between diets suggests that this parameter may be important in fish nutritional investigations.     

Effect of protein source and nutrient density on growth efficiency, histology and plasma amino acid concentration of rainbow trout (Oncorhynchus mykiss Walbaum)

The effects of protein source and nutrient density on growth efficiency, nutrient digestibility and plasma amino acid concentrations of rainbow trout were evaluated. A 3 by 2 factorial treatment design with three protein sources, fish meal–barley (F–B), plant concentrates (PC) and plant meals (PM), and two nutrient densities were used. A commercial reference diet was also fed. Triplicate tanks of 30 fish (initial wt. 28 g) were fed each diet, and the final weight averaged 240 g fish⁻¹. Protein source and nutrient density affected feed intake, weight gain and feed conversion ratio. Weight gain of trout fed the PC and PM diets was approximately 10% less than fish fed the F–B diets. Protein retention was affected by protein source, but not nutrient density, and was the highest for the fish fed diets containing fish meal and the lowest for the fish fed PM diets. Apparent digestibility coefficients and apparent amino acid availabilities of the diets corresponded with differences in weight gain. This study provides further evidence that growth rates of trout fed fish meal‐free diets, using conventional and concentrated plant protein ingredients, are good but some limitation to growth exists in the fish meal‐free diets.     

Lipid-stimulated somatostatin secretion in rainbow trout,Oncorhynchus mykiss

Previous work has shown that somatostatins (SS) affect teleost lipid metabolism indirectly by inhibition of insulin (INS) and directly by stimulation of hepatic lipolysis. In the present study, rainbow trout (Oncorhynchus mykiss) were used to characterize further the lipid-SS relationship by evaluating how lipid, contributes to SS secretion bothin vivo andin vitro. In vivo hyperlipidemia was induced for up to 3 h by short-term (2 min) infusion of a triacylglycerol (TG)-rich lipid emulsion (20% Intralipid^®). Plasma total lipid concentration increased 118 and 155% over control levels 1 h and 3 h, respectively, after infusion; much of this increase was due to elevated plasma fatty acids (FA), which increased 39 and 520%, respectively, over the same time-frame. The hyperlipidemic pattern was attended by a significant increase in the plasma concentration of SS. The specific effects of fatty acids were evaluated on isolated Brockmann bodies. Palmitic acid and oleic acid stimulated SS release 378 and 82%, respectively, over baseline levels. These results indicate that lipids, and in particular fatty acids, modulate SS secretion in rainbow trout.     

Identification of a novel gill-specific calpain from rainbow trout (Oncorhynchus mykiss)

Calpains are calcium-dependent neutral proteases responsible for many cellular functions. The two forms of calpain ubiquitously expressed in mammalian tissues are known as μ-calpain and m-calpain. We report here the identification of a novel calpain that is similar to but distinct from the μ- and m-calpains in rainbow trout. The cDNA of the novel gene is 2623 bp in length with a single open reading frame. The predicted protein (676 amino acids) contains the conserved calpain characteristic domains that include: domain I (pro peptide), II (cysteine catalytic site), III (electrostatic switch), and IV (calmodulin-like) with five Ca²⁺-binding EF hands. Northern blot and RT-PCR analyses demonstrated that the novel calpain gene is predominantly expressed in rainbow trout gills. Comparison of the novel protein with the ubiquitously expressed calpains and several mammalian tissue-specific calpains revealed that the novel calpain is an orthologue of the mammalian digestive tract specific calpain (calpain 9).     

Total calcium and inorganic phosphate in the blood plasma of farmed rainbow trout, Oncorhynchus mykiss

Reference (physiological) value intervals determined by the lower 2.5% and upper 97.5% quantiles were calculated for total calcium (Ca _t ) and inorganic phosphate (P) in farmed rainbow trout, Oncorhynchus mykiss , at an age of 10–12 months in raceway culture. Total calcium levels were significantly ( P ≤0.0009) greater in males (2.63–4 mmol L⁻¹, n =47, mean weight 359±142 g) than in immature females (2.48–3.97 mmol L⁻¹, n =410, mean weight 400±145 g). The reference range for P in immature females and males was 3.09–9.32 mmol L⁻¹ ( n =467, mean weight 401±144 g). The distribution and density of the quantiles in the tested reference group were made possible by the use of histograms, which showed normal distributions for Ca _t in males but not for Ca _t and P in immature females and males, in which a sinistral asymmetry was found.     

Nitrogen utilisation and ureogenesis as affected by dietary nucleic acid in rainbow trout (Oncorhynchus mykiss) and turbot (Psetta maxima)

We evaluated the influence of dietary nucleic acid (NA) supplied either as an RNA extract (RNA) or as brewer's yeast (BY) on nitrogen (N) utilisation, N excretion and ureogenesis in a freshwater teleost, the rainbow trout (Oncorhynchus mykiss) and a marine teleost, the turbot (Psetta maxima). Five diets containing two levels of NA or BY were formulated for each species, and fed to juvenile rainbow trout (initial body weight (IBW): 21.7 g) and turbot (IBW: 11.8 g) over 8–10 weeks. Besides growth and N utilisation, we monitored total ammonia-N and urea-N excretion rates and measured the activities of selected enzymes (glutamate dehydrogenase, ornithine carbamoyltransferase, arginase and uricase). There was no clear N sparing effect of dietary NA or BY supplementations in either species. Feeding diets containing the RNA extract led to an almost two-fold increase in postprandial plasma urea-N and uric acid concentrations and in urea-N excretion rates. Glutamate dehydrogenase and arginase activities were decreased in fish fed NA supplemented diets. Uricase activities were inversely related to dietary NA levels. Data on N excretion and enzyme activities clearly suggest that ureogenesis is influenced by dietary NA in both species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Improvement of flesh quality in rainbow trout (Oncorhynchus mykiss) fed supranutritional dietary selenium yeast is associated with the inhibited muscle protein degradation

Supranutritional dietary selenium (Se) has been demonstrated beneficial for fish health, however, its effects on fish flesh quality remain unclear. This study investigated the effects of supranutritional dietary Se on the flesh quality of rainbow trout (Oncorhynchus mykiss) and their potential mechanism. Fish were fed a basal diet supplemented with or without graded Se yeast for 10 weeks. Results showed that Se supplementation significantly increased fillet crude protein and enhanced fillet water‐holding capacity as well as fillet firmness. Supplementing with both 2 and 4 mg/kg Se significantly downregulated the expressions of two autophagy–lysosome‐related genes (autophagy‐related 12‐like and gamma‐aminobutyric acid type A receptor‐associated protein‐like 1) in fish muscle, while supplementing with 4 mg/kg Se also significantly downregulated the expression of two ubiquitin–proteasome‐related genes (muscle RING finger 2 and F‐box protein 25). Correlation analysis indicated that the improved fillet quality parameters were closely correlated with the expressions of these differentially expressed genes. This study revealed that dietary Se was effective for the improvement of rainbow trout flesh quality, and the improved fish flesh quality was associated with the inhibited protein degradation in fish muscle.