Dietary carbohydrate and growth,body composition and heat increment in rainbow trout (Salmo gairdneri)

A study was conducted to determine growth, body composition and heat increment (HI) of rainbow trout reared on isonitrogenous high digestible carbohydrate (HC) and high fat (HL) diets at 15°C. Trout reared on the HC diet had a significantly lower final body weight and carcass lipid content and a significantly higher feed:gain ratio and carcass protein content than trout reared on the HL diet after 12 weeks on the test diets. The lower carcass lipid composition indicates that trout do not readily convert dietary carbohydrates into fat. HI of trout reared on the HC diet was significantly higher than that of fish reared on the HL diet. Considering the poor utilization of dietary carbohydrates as an energy source and the apparently low conversion of dietary glucose into fat, the cause of the higher HI associated with the HC diet is not completely understood. However, it is possible the elimination of excess dietary glucose is an energy dependent process resulting in an increased heat production which would partly account for the increased HI. Furthermore, if the HC diet is a low net energy diet, then the metabolism and utilization of dietary and tissue proteins (amino acids) for energy by the trout may also be partly responsible for the increased HI of the trout reared on the HC diet.     

Dynamics of plasma free amino acids in rainbow trout (Oncorhynchus mykiss under variety of dietary conditions

Plasma amino acid profiles in the systemic circulation of rainbow trout acclimated to diets containing different protein sources, with and without supplementation with free amino acids, were monitored for up to 120 hours after consumption of the diets. Plasma concentrations of lysine, methionine, and isoleucine increased more rapidly after feeding a diet supplemented with these amino acids in free form and, depending upon the increase in plasma concentration, remained at concentrations above those in fish fed the basal diet for at least 24h after feeding. Dietary supplementation with isoleucine increased plasma concentrations of leucine and valine as well as of isoleucine. Maximum plasma concentrations for most amino acids were attained between 12 and 24h postprandial. Dietary inclusion of gelatin caused more rapid elevations in plasma glycine and serine than did free glycine. Feeding at three hour intervals resulted in stable plasma concentrations of individual amino acids in contrast to the fluctuations occurring when fish were fed once daily. Frequent feeding favoured a higher proportion of protein to lipid in body composition.     

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.     

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.     

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.     

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.     

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.     

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.     

Apparent amino acid availability from feedstuffs in extruded diets for rainbow trout Oncorhynchus mykiss

Apparent amino acid availability coefficients of several typical and novel feed ingredients were determined in rainbow trout using extruded diets and the faecal stripping technique. The ingredients were tested included five fish meals, three terrestrial animal by‐products, five plant protein concentrates, four plant meals, and seven low‐protein plant ingredients. Amino acid availability from the fish meals was relatively high ranging from 90 to 101%. Lower coefficients overall were observed for Menhaden fish meal FAQ when compared to the other fish meals. No differences in apparent amino acid availability were detected among the animal by‐products. Within the plant concentrate group, rice protein concentrate and barley protein concentrate exhibited generally lower amino acid availabilities compared to other concentrates tested. Among the plant meals, only the availabilities of histidine, valine, isoleucine and lysine in flaxseed meal were lower than those of soybean meal. Apparent amino acid availabilities among the low‐protein plant products were variable and significantly different.     

Metabolism of exogenous histamine in rainbow trout (Oncorhynchus mykiss)

Information on the metabolism of exogenous histamine in fish is of much concern regarding the effect of dietary histamine on fish. Histamine catabolic enzymes, diamine oxidase and histamine N-methyl transferase were measured in the tissues of rainbow trout. Diamine oxidase was detected in the stomach, pylorus caeca and intestine. Histamine N-methyl transferase was detected only in the liver.A change in the contents of histamine and its metabolites was observed in the tissues of rainbow trout after oral administration of histamine. A large amount of imidazole acetic acid was observed in the serum, kidney, liver and muscle. On the other hand, 1-methyl histamine was detected only in the liver. Histamine and its metabolites, imidazole acetic acid and 1-methyl histamine were metabolized and diminished within 48 hr in all tissues.These results showed that histamine was metabolized by two metabolic routes in rainbow trout. One is the main pathway producing imidazole acetic acid by intestinal diamine oxidase and the other is the complementary pathway producing 1-methyl histamine by liver N-methyl-transferase.     

Dietary formic acid enhances apparent digestibility of minerals in rainbow trout, Oncorhynchus mykiss (Walbaum)

The effect of dietary formic acid on the availability of phosphorus (P) from a fishmeal-based diet by rainbow trout, Oncorhynchus mykiss (Walbaum), reared in brackish water (5–6 g L⁻¹) was determined. Diets containing a low amount of P (6.0 mg P g⁻¹ dry matter) were acidified with 0, 4 and 10 mL kg⁻¹ formic acid and fed to trout (average weight, 520 g) for 4 weeks. The measured pH of the diets were 6.3, 5.8 and 5.3, respectively. The intestinal solubility of P and the digestibility of P were measured by stripping the faecal matter from the fish. The apparent digestibility coefficient (ADC) of P significantly ( P < 0.05) increased from 69.5% to 75.0% of the basal diet in fish fed diets containing 10 mL kg⁻¹ formic acid. The solubility of P in the intestine was highly variable within each treatment, and the differences were not significant. The pH of intestinal content increased with the increase in dietary formic acid concentration. The ADC of magnesium and calcium also showed a significant ( P < 0.05) increase with the acidification of diet by formic acid.     

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).     

Effects of traditional rainbow trout (Oncorhynchus mykiss) breeding on dissolved organic nitrogen pools and microbial activity in the water

Seasonal effects of rainbow trout (Oncorhynchus mykiss) breeding in traditional, flow‐through ponds on abundance and activity of bacteria, and on pools and cycling of dissolved free and combined amino acids (DFAA and DCAA), were studied in three aquaculture systems in Denmark. The fish breeding was found to increase both abundance and production of bacteria by about 60% (annual average of all three systems). The bacterial production correlated positively with temperature, and was 10 to 60‐fold higher in summer than winter. Concentrations of DFAA (range: 45–690 nM) increased by 40–350% (mean of 89%) during passage in the systems. The bacterial uptake of DFAA was stimulated by 42% (annual average value of the three systems) between inlets and outlets. Uptake of DFAA was estimated to sustain from 45% (C) and 70% (N) of the bacterial C and N demand. Concentrations of DCAA, e.g., proteins and peptides, varied from 600 to 3900 nM, and increased on average 89% during passage in the three systems. Bacterial capacity for utilization of DCAA was determined as leucine peptidase activity and related to the bacterial density. The cell‐specific peptidase activity was, on average, 103% higher at outlets than inlets. Changes in the peptidase activity did not correlate with variations in DCAA pools or the bacterial production. Concentrations of ammonium, urea and phosphate increased on average by 88%, 103% and 66%, respectively, during passage in the three systems, whereas no changes were observed for nitrate. Our study shows that traditional fish breeding impacts pools and cycling of dissolved organic N, as well as bacterial dynamics. The short residence time of water in the ponds (3–6 h) implies that most of the nutrients produced in the systems were discharged into the downstream water, rather than assimilated by bacteria within the systems.     

The role of cortisol in amino acid mobilization and metabolism following exhaustive exercise in rainbow trout (Oncorhynchus mykiss Walbaum)

The role of cortisol in the mobilization and metabolism of amino acids following exhaustive exercise in rainbow trout (Onorhynchus mykiss) was investigated. Plasma cortisol levels were elevated for 4h following exercise and 6, of a total of 22 amino acids, showed significant responses. In white muscle, alanine and glutamine were elevated immediately after exercise through to 4h, whereas glutamate, and the branched-chain amino acids (BCAA), isoleucine, leucine and valine, all decreased over this time. In plasma all of these amino acids increased from 2-4h post-exercise, while in liver, alanine and glutamine increased, glutamate did not change and the BCAAs declined over this time. Blockade of the post-exercise elevation in plasma cortisol with either metyrapone (cortisol synthesis inhibitor) or dexamethasone (cortisol release blocker) prevented the changes in tissue amino acid levels. This study demonstrates that cortisol can act rapidly (within minutes to hours) to alter amino acid metabolism in fish. A model is presented to explain the action of cortisol on amino acid metabolism.     

Assessing the bioavailability of L‐methionine and a methionine hydroxy analogue (MHA‐Ca) compared to DL‐methionine in rainbow trout (Oncorhynchus mykiss)

Different synthetic forms of supplemental methionine (Met) are produced commercially by various manufacturers. Limited information exists on the bioavailability of these different forms in commercially relevant fish species. The objective of this study was to compare the relative bioavailability of L‐methionine and a methionine hydroxy analogue calcium salt (MHA‐Ca) to DL‐methionine in a commercially relevant species, rainbow trout, using a linear slope‐ratio assay. In addition, the ability of the rainbow trout to effectively convert the D‐isomer of methionine into the biologically active L‐isomer was studied by comparing the relative bioavailability of L‐methionine to DL‐methionine. Nine (9) diets were created from a methionine‐deficient basal diet which was supplemented with graded equimolar levels of methionine (0.1%, 0.2%, 0.3% of diet by weight) from either DL‐methionine, L‐methionine and methionine hydroxy analogue calcium salt (MHA‐Ca). Diets were fed in triplicate (IBW = 24.0 ± 0.6 g) for 12 weeks using a pair‐feeding protocol. The basal diet was formulated to be deficient in both methionine (0.5%) and cysteine (0.3%), whereas meeting all other known nutritional requirements of rainbow trout. MHA‐Ca was found to be less available (P < 0.05) than DL‐methionine with relative bioavailability values of 69%, 60% and 73% based upon weight gain, growth rate (TGC) and retained nitrogen values respectively. No significant differences (P > 0.05) in bioavailability were found between DL‐methionine and L‐methionine. These findings lead to the conclusion that differences in bioavailability do exist between sources of synthetic methionine, additionally findings suggest that rainbow trout are able to effectively convert the D‐isomer of methionine into the L‐isomer without significantly affecting fish performance.     

Effects of egg aging on the metabolites of ovarian fluid in rainbow trout,Oncorhynchus mykiss

Nuclear magnetic resonance ‐based metabolomics was applied to study effects of egg aging on ovarian fluid metabolites in rainbow trout, Oncorhynchus mykiss. The eggs of three females were pooled and then assigned to three plastic vials for 18 days in vitro storage at 4°C. Ovarian fluid samples were taken 0, 6, 12 and 18 days after storage. Three groups of metabolites including amino acids, osmolytes and energy metabolites were found to change during storage period. The glucose levels of ovarian fluid showed significant decreases on days 12 and 18 after storage (p < .05). For acetoacetate and acetate, significant increases were observed, respectively, on days 12 and 18 after storage (p < .05). The creatine levels of ovarian fluid increased significantly on days 12 and 18 after storage (p < .05). Lactate levels in ovarian fluid elevated during storage period (p < .05). Glycerol levels showed a significant increase on days 12 and 18 after storage (p < .05). The values of osmolytes in ovarian fluid (e.g., betaine, taurine, trimethylamine‐N‐oxide, N,N‐dimethylglycine) showed a decreasing trend on day 12 which continued until the end of storage on day 18 (p < .05). Almost all amino acids elevated on days 12 and 18 after storage (p < .05). After an apparent elevation in isoleucine levels on days 6 and 12, this amino acid decreased on day 18 after storage (p < .05). The osmolytes might act as antioxidant against free‐radicals produced as a result of over‐ripening. Glucose can be used as energy resource for eggs and bacteria during ova storage. Also, change pattern of amino acids indicate hydrolysis of proteins as the time of storage increases.     

The uptake and esterification of radiolabelled fatty acids by enterocytes isolated from rainbow trout (Oncorhynchus mykiss)

To study the intestinal fatty acid absorption in fish in vitro, enterocytes were isolated from the intestine of rainbow trout and incubated with an equimolar mixture of seven fatty acids [16:0, 18:1n-9, 18:2n-6, 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3] in which the component carrying a radioactive label was varied. The fatty acid mixture was presented in the form of micelles formed by sonication with sodium taurocholate. Control studies showed that the presence of sodium taurocholate in the incubation medium caused an immediate 23% increase in the mortality of the cells but the remaining cells were viable. The effect of the bile salt on cellular permeability was evident at longer exposure periods. The proportions of 14C-labelled 20:5n-3 and 22:6n-3 present as monomeric fatty acids in the micellar solutions were higher than those of 16:0, 18:1n-9, 18:2n-6, 18:3n-3 and 20:4n-6. The rates of uptake of 14C-labelled 20:4n-6, 20:5n-3 and 22:6n-3 by enterocytes were significantly lower than those of the other fatty acid substrates over the first minute of incubation but no significant differences in uptake rate between fatty acids were obvious over a 15 min incubation period. Notable differences were observed between substrates in their distribution pattern in enterocyte lipid classes. Although most of the radioactivity from all radiolabelled substrates was recovered in triacylglycerols, the amounts of 14C- labelled 16:0, 20:4n-6, 20:5n-3 and 22:6n-3 recovered in the polar lipid fraction were higher than those of 14C-labelled C18 substrates, particularly after 15 min. Conversely, the initial esterification rates of 18:1n-9, 18:2n-6 and 18:3n-3 into triacylglycerols were significantly higher than those of 20:4n-6, 20:5n-3 and 22:6n-3. It is concluded that isolated enterocytes can be used for the study of the mechanism of intestinal fatty acid absorption in fish.     

Dietary effects on insulin and glucagon plasma levels in rainbow trout (Oncorhynchus mykiss) and gilthead sea bream (Sparus aurata)

The effects of dietary amino acid profile (based on muscle (M) or whole body composition (WB) and the balance between indispensable (IAA) and dispensable amino acids (DAA) in the diet, on plasma levels of insulin and glucagon, were analysed in rainbow trout and gilthead sea bream. Plasma insulin values (baseline and 6 h postfeeding) were higher in trout than in sea bream, but the relative postfeeding increase was more pronounced in sea bream. Within the same dietary amino acid profile, diets with lower IAA/DAA, had a lower effect on the postfeeding secretion of insulin in both species. Circulating levels of glucagon (baseline and postfeeding relative increases) were higher in sea bream. In trout, diets with WB amino acid profile had a greater secretory effect on postfeeding glucagon than did diets with M profile, while gilthead sea bream showed an inverse response to circulating glucagon with respect to diet. Muscle insulin and insulin growth factor-I binding parameters were not affected by the dietary regimen. The postfeeding glucagon response depends on both the dietary AA profile and the fish species, while that of insulin seems to be more uniform, and is affected in a similar way regardless of the species.     

Partial or total replacement of fishmeal by solvent-extracted cottonseed meal in diets for juvenile rainbow trout (Oncorhynchus mykiss)

The effect of solvent‐extracted cottonseed meal (SCSM) as a partial or total replacement of fishmeal was studied in juvenile rainbow trout (Oncorhynchus mykiss). Six experimental diets SCSM0, SCSM25, SCSM50, SCSM75, SCSM75A and SCSMT, containing a gradient of SCSM 0, 152, 305, 465, 460 and 610 g kg^?1 to replace 0, 112.5, 225, 337.5, 337.5 and 450 g kg^?1 fishmeal protein were fed to triplicate groups (initial body weight of 39.2 ± 0.1 g) for 8 weeks. The diet SCSM75A was supplemented with lysine and methionine, to be similar to SCSM0 for juvenile rainbow trout. Faeces were colleted after 4 weeks of normal feeding for apparent digestibility coefficients (ADC) of dry matter, crude protein and gross energy determination. Total replacement of fishmeal adversely affected growth performance. Fish fed with diet SCSMT had significantly (P < 0.05) lower weight gain, specific growth ratio, feed conversion efficiency (FCE) and protein efficiency ratio than fish fed with other diets. The FCE of SCSM75 and SCSM75A were significantly lower (P < 0.05) than those of fish fed with SCSM0 diets. The ADC of the dry matter of SCSM75 and SCSMT were significantly lower than the SCSM0 diet, and the ADC of crude protein and the energy of SCSMT were the lowest (P < 0.05). The ADC of threonine, proline, alanine, valine, isoleucine, leucine, lysine and methionine of fish fed with diet SCSMT were lower. Lysine and methionine supplement positively affected the ADC of SCS75A diet. There were no significant differences in the fish body composition. It is shown that SCSM can be utilized in the juvenile rainbow trout diet up to 305 g kg^?1, to replace about 50% of fishmeal protein in this experiment.