Effects of ration level in rainbow trout, Oncorhynchus mykiss (Walbaum), on sensory characteristics, lipid content and fatty acid composition

Effects of feed ration levels (RLs) and age on sensory characteristics, fillet lipid content (LC) and fatty acid composition (FA) of rainbow trout, Oncorhynchus mykiss (Walbaum), fillet were investigated in a longitudinal feeding experiment. Daily RLs were administered as a percentage of the ration size necessary for expected maximum growth (RL100). Fish on RL50, RL75, RL100 and RL200 were sampled at 1.4, 2.0 and 2.4 years of age. Analyses of LC and FA were carried out on a fillet cross-section. Sensory analyses were performed by an expert panel using ranking and conventional profiling. RL had no effect on the intensity of fresh smell or any sensory characteristic of taste or consistency of fish on RLs over 75%, irrespective of age; fish on RL50, however, scored significantly lower for fresh taste and firmness. Age had no effect on the intensity of total taste, whereas fresh taste increased and rancidity decreased with increasing age irrespective of RL. LC related strongly to RL and age, and increased up to 2.0 years of age. An increase in LC resulted in a higher level of monounsaturated FA and a lower level of polyunsaturated FA. A restriction to RL75 had no negative effects on sensory characteristics, LC or FA of the fillet.     

Influence of dietary palm oil on growth, tissue fatty acid compositions, and fatty acid metabolism in liver and intestine in rainbow trout (Oncorhynchus mykiss)

This study aimed to investigate the effects of dietary crude palm oil (CPO) on fatty acid metabolism in liver and intestine of rainbow trout. Triplicate groups of rainbow trout for 10 weeks at 13 °C were fed on diets in which CPO replaced fish oil (FO) in a graded manner (0–100%). At the end of the trial, fatty acid compositions of flesh, liver and pyloric caeca were determined and highly unsaturated fatty acid (HUFA) synthesis and fatty acid oxidation were estimated in isolated hepatocytes and caecal enterocytes using [1‐¹⁴C]18:3n‐3 as substrate. Growth performance and feed efficiency were unaffected by dietary CPO. Fatty acid compositions of selected tissues reflected the dietary fatty acid composition with increasing CPO resulting in increased proportions of 18:1n‐9 and 18:2n‐6 and decreased proportions of n‐3HUFA, 20:5n‐3 and 22:6n‐3. Palmitic acid, 16:0, was also increased in flesh and pyloric caeca, but not in liver. The capacity of HUFA synthesis from 18:3n‐3 increased by up to threefold in both hepatocytes and enterocytes in response to graded increases in dietary CPO. In contrast, oxidation of 18:3n‐3 was unaffected by dietary CPO in hepatocytes and reduced by high levels of dietary CPO in enterocytes. The results of this study suggest that CPO can be used at least to partially replace FO in diets for rainbow trout in terms of permitting similar growth and feed conversion, and having no major detrimental effects on lipid and fatty acid metabolism, although flesh fatty acid compositions are significantly affected at an inclusion level above 50%, with n‐3HUFA reduced by up to 40%.     

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.     

Effects of dichlorvos and formalin on fatty acid metabolism of rainbow trout (Oncorhynchus mykiss) skin cells in primary culture

The effects of sub-lethal doses of dichlorvos and formalin, antimicrobial/parasitic agents used in aquaculture, on lipid composition and metabolism of rainbow trout skin cells in primary culture were investigated. [1-¹⁴C]Stearic (18:0), [1-¹⁴C]lin 18:2n-6) and [1-¹⁴C]linolenic (18:3n-3) acids were used as tracers to determine effects on fatty acid incorporation and metabolism. Formalin increased cell numbers and reduced the lipid content of the cells and the incorporation of radioactive fatty acids. The effects of dichlorvos were qualitatively similar but quantitatively less. Formalin induced relatively small but significant changes in lipid class composition including a decreased proportion of phosphatidycholine with increased proportions of sphatidylethanolamine and phosphatidylserine. Dichlorvos had no significant effect on lipid class compositions. The trout primary skin cells expressed substantial 9, 6 and 5 fatty acyl desaturase activities. Although, as expected, the cells were m active towards [1-¹⁴C]18:3n-3, the cells were unusually active towards [1-¹⁴C]18:2n-6. Both dichlorvos and, especially, formalin appeared to significantly inhibit 9 and 6 desaturation. Changes in the distribution of radioactivity between individual spholipid classes was also influenced by formalin and dichlorvos, and this may be related to changes in desaturase activity. This study has shown that topically active agents used in aquaculture, formalin and dichlorvos, had a range of effects on the rainbow trout skin cell cultures that may affect cell proliferation and lipid and fatty acid metabolism. Both agents significantly inhibited desaturation of fatty acids, particularly of 18:2n-6 to 20:4n-6 and, as 20:4n-6 is a major eicosanoid precursor ish and considering the importance of eicosanoids in the biochemistry of skin, it is suggested that these agents may have direct effects on fish skin that could have important consequences for fish health in general.     

The effect of culture on morphology, lipid and fatty acid composition, and polyunsaturated fatty acid metabolism of rainbow trout (Oncorhynchus mykiss) skin cells

Rainbow trout (Oncorhynchus mykiss) skin cell cultures were obtained by trypsinization of the tissue and grown in Leibovitz L-15 medium. Lipid class compositions, and fatty acid profiles of total lipids and individual phospholipid classes were determined at different times of culture. The metabolism of polyunsaturated fatty acids (PUFA) was investigated by incubating primary cultures after 7 and 14 days with [1-¹⁴C]18:2n-6 and [1-¹⁴C-]18:3n-3. The change in morphology between epithelial-like primary cultures and fibroblastic-like secondary subcultures was accompanied by alterations in the lipid composition. Polar lipids became predominant by 14 days in culture. The relative proportions of phosphatidylcholine (PC), the most abundant phospholipid, phosphatidylinositol and cholesterol increased significantly, while sphingomyelin decreased. Saturated fatty acids, 18:1n-9, n-6 and n-9PUFA were more abundant in total lipid in cultures at 14 days and 4 months than in cells initially isolated which contained higher percentages of longer chain monoenes and n-3PUFA. The changes in fatty acid composition with time in culture were observed in all the major phospholipid classes. Rainbow trout skin cells in culture desaturated and elongated both 18:2n-6 and 18:3n-3, with 20:4n-6 and 20:5n-3 being the most abundant products, respectively. PC presented the highest incorporation of radioactivity, especially following incubation with 18:3n-3. Lipid metabolism in general increased with the age of primary cultures, with both the amount of C18 PUFA incorporated and metabolized by desaturation/elongation significantly increased in 14 day cultures compared to 7 day cultures. Product/precursor ratios calculated for both n-6 and n-3 fatty acids showed that, while 6 desaturase activity was increased significantly with cell age, 5 desaturase activity was more affected by the fatty acid series, with 18:3n-3 being more readily transformed to 20:5n-3 than 18:2n-6 to 20:4n-6. Further desaturation of 20:5n-3 to hexaenes was low. Overall, the data suggested that the trout skin cell cultures were more similar to mammalian skin fibroblasts than mammalian epidermal/keratinocyte cultures.     

Relationships between growth and disease resistance in rainbow trout,Oncorhynchus mykiss (Walbaum)

Rainbow trout from 23 families were evaluated for growth and resistance to the bacterial coldwater disease (BCWD) caused by Flavobacterium psychrophilum and infectious haematopoietic necrosis (IHN) caused by IHN virus. Average family weights were between 161 and 263 g with an average of 225 g at 213 days post‐fertilization with specific growth rates ranging from 2.37 to 2.88. Per cent survival of fish challenged with F. psychrophilum was between 18% and 100%, while for those challenged with IHNV, the range was between 12% and 93%. Significant positive correlations were found for end body weight and resistance to IHN (P < 0.05) and for early body weight and resistance to BCWD (P < 0.1). However, no significant correlations were detected between resistance to both pathogens or disease resistance and overall genetic diversity or diversity within the major histocompatibility locus.     

Use of probiotics to control furunculosis in rainbow trout, Oncorhynchus mykiss (Walbaum)

Aerobic heterotrophic bacteria were isolated from the intestinal contents of Atlantic salmon, Salmo salar , rainbow trout, Oncorhynchus mykiss , and turbot, Scophthalmus maximus , on tryptone soya agar and De Man Rogosa and Sharpe agar, of which 11 of 177 (6% of the total) of the isolates were antagonistic to Aeromonas salmonicida . Four of these cultures, which were identified tentatively as A. hydrophila , Vibrio fluvialis , Carnobacterium sp. and an unidentified Gram-positive coccus, were beneficial to fish when fed singly or as an equi-mixture. Feed supplemented with the putative probiotics indicated survival of the organisms in the gastrointestinal tract for 7 days. Feeding with the probiotics for 7 and 14 days led to better survival following challenge with A. salmonicida . There was no indication of serum or mucus antibodies to A. salmonicida , but there was an increased number of erythrocytes, macrophages, lymphocytes and leucocytes, and enhanced lysozyme activity in the fish.     

Effects of supplemental dietary l‐carnitine and ractopamine on the performance of juvenile rainbow trout,Oncorhynchus mykiss

This study was carried out to investigate a possible protein‐sparing action of l ‐carnitine and ractopamine in rainbow trout, Oncorhynchus mykiss. An 8‐week feeding trial was carried out to evaluate the effects of supplementation of three levels of l ‐carnitine (0, 1 and 2 g kg^?1) and two levels of ractopamine (0 and 10 mg kg^?1) on growth performance, fillet fatty acid compositions and blood biochemical parameters in a 3 × 2 factorial experimental design. Ractopamine and 1 g kg^?1 carnitine improved the specific growth rate (1.03% and 1.05% day^?1), feed conversion ratio (FCR, 1.3 and 1.29), protein efficiency ratio (PER, 1.88 and 1.85) of fish and crude protein (73.5 and 73.8) content of fish fillet. l ‐carnitine and ractopamine increased the levels of albumin, total protein and globulin in the serum of fish. Apart from eicosapentaenoic acid and docosahexaenoic acid, other fatty acids of fish fillet were increased by ractopamine, while total saturated fatty acids were almost intact. However, the total n‐3 poly unsaturated fatty acids were reduced by l ‐carnitine supplementation (P<0.05). The present study showed that 1 g kg^?1l ‐carnitine and 10 mg kg^?1 ractopamine each can improve the performance of rainbow trout and their combination in diet could enhance the protein level and change the fatty acids profile in fillet muscle.     

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.     

Influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum)

The influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum), was studied experimentally in the laboratory and at a fish farm. In experiment I, the effect of parasitic infection on columnaris disease was studied using F. columnare carrier fish. The fish were exposed to Diplostomum spathaceum cercariae and a set of other stressors in order to induce clinical columnaris infection. Parasitic infection and other stressors failed to induce the disease. Disease occurred when the fish were challenged with F. columnare, but D. spathaceum infection did not enhance the severity of the infection. In experiment II, the influence of rearing density and water temperature was studied. Overall mortality was highest in fish at normal rearing density with high temperature (+23 degrees C). At low temperature (+18 degrees C) mortality was not affected by rearing density, but the transmission of columnaris disease was faster at normal rearing density at both temperatures. This supports the view that reduction of fish density could be used in prevention of columnaris disease especially if water temperature is high. Because the lower rearing density can also decrease the transmission of ectoparasites and penetrating endoparasites, it could be an efficient tool in ecological disease management.     

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.     

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.     

Inclusion of macroalgae meal (Macrocystis pyrifera) as feed ingredient for rainbow trout (Oncorhynchus mykiss): effect on flesh fatty acid composition

The use of macroalgae as an additional component in animal feeding has been studied. However, information on how it could influence muscle composition of fish body is scarce. This study evaluates four diets with different macroalgae inclusion levels (0%, 1.5%, 3% and 6%) to test the effect on body fatty acid composition of rainbow trout. Tanks with a volume of 600 L were stocked with 60.6 ± 7.9 g fish at a density of 45 individuals tank^?1 and fed for 124 days. At the end of the experiment there were not significant differences (P<0.05) in muscle proximate composition among fish fed the different treatments. However, it was determined that inclusion of 3% and 6% of macroalgae meal resulted in a significant increase (P<0.05) of polyunsaturated fatty acids (PUFAs) in muscle. In summary, macroalgae meal in rainbow trout diets do not enhance the quantity of protein and lipid contents at muscle level but an addition of 3–6% might contribute to increase the level of PUFAs, specially EPA, DHA and LIN. Thus, use of macroalgae meal might help to increase lipid quality content in the final product due the beneficial effects of PUFAs for human health.     

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.     

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.