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Aquaculture is one of the most thriving animal production sectors, and Nile tilapia (Oreochromis niloticus) farming represents 8% of total finfish culture. However, the industry sustainability depends on the development of cost‐effective and environmental friendly feeds. This study aimed to reduce dietary protein levels in diets for juvenile tilapia and to minimize diet environmental impact while maximizing biological efficiency. A growth trial was performed using five isoenergetic plant protein‐based diets with decreasing levels of crude protein: 360, 340, 320, 300 and 280 g/kg diet (D360, D340, D320, D300 and D280, respectively). Dietary protein utilization was assessed by metabolic trials using a radiolabelled amino acid mixture. Tilapia in all treatments showed similar growth performance and feed intake. Feed conversion ratio was significantly higher in fish fed the D280 than the D360 diet, while no differences were found for other treatments. Protein retention was significantly higher in tilapia fed the D300 than the D360 diet. Amino acid catabolism increased in fish fed the D360 diet, though without significant differences in muscle amino acid retention. This study demonstrates that dietary protein levels can be reduced to 300 g/kg diet without hindering tilapia growth and feed conversion ratio, while reducing environmental nitrogen losses. FCT: UID/Multi/04326/2019/ IF/00482/2014/CP1217/CT0005/ DL57/2016/CP1361/CT0033. info:eu-repo/semantics/publishedVersion

Recent studies reported positive effects of taurine supplementation to plant protein-based diets on fish growth performance and feed conversion ratio but the mechanisms involved remain unclear. The objective of this work was to provide a first insight on the importance of taurine on dietary protein and lipid absorption and utilisation, in Senegalese sole (Solea senegalensis) juveniles fed plant-based diets. Seven practical diets were tested: a fishmeal-based diet (FM) and a plant protein-based formula (PP85) to which taurine was added in graded levels: 0.2% (TAU1), 0.3% (TAU2), 0.4% (TAUS), 0.7% (TAU4) and 1.5% (TAU5). Short-term metabolic trials were performed by tube-feeding the different experimental diets labelled with different tracers: C-14-taurine, C-14-triolein (as tracer of dietary triacylglycerol) and a mixture of C-14-amino acids. Feeding sole juveniles with the PP85 diet affected taurine metabolism, since a higher proportion of dietary taurine was directed to gallbladder and probably used for bile salt synthesis. Consistent with this, fish receiving the plant-based low taurine-content diets (PP85 and TAU1) displayed a poor triacyglycerol digestion capacity. A higher proportion of C-14-amino acids was retained in fish muscle when fed the TAU5 compared to the PP85 diet. These short-term trials suggested that taurine supplementation to a plant-based diet might be beneficial to sole performance by enhancing bile salt synthesis and triacylglycerol digestion as well as muscle amino acid retention. However, this dietary supplementation should be tested in long-term trials to verify the effect on fish growth and to assess the beneficial effects on protein and lipid metabolism suggested here. Statement of relevance in the general field of aquaculture (limited to 60 characters): Adding taurine to a plant-based diet is beneficial for sole (C) 2016 Elsevier B.V. All rights reserved. National Funds through Fundacao para a Ciencia e Tecnologia [PTDC/MAR/117047/2010] Fundacao para a Ciencia e Tecnologia, Portugal [SFRH/BDP/65578/2009] info:eu-repo/semantics/acceptedVersion

The present study aimed to assess the effects of increased availability of dietary amino acids (AA) on brain monoamine neurotransmitters and the metabolic processes resulting from stressful situations in fish. Senegalese sole ( Solea senegalensis ) juveniles (24.2 ± 0.4 g wet mass) were weekly subjected to an acute handling stressor (HDLG) or remained undisturbed (CTL). Additionally, both treatments were fed a control or a high protein (HP) diet (CTL, CTL HP, HDLG and HDLG HP). The HP diet slightly increased the levels of digestible indispensable AA, together with tyrosine and cysteine. Repeated handling induced a stress response after 14 and 28 days in fish held at both HDLG and HDLG HP treatments. While dietary treatment and handling stress activated the serotonergic system at 14 days, these effects were not observed after 28 days. In addition, the HP diet minimized the decrease in plasma indispensable AA due to repeated handling stress after 28 days. It was concluded that HP diet decreased post-stress plasma glucose and lactate levels in HDLG HP specimens only at 14 days of treatment. Moreover, dietary treatment was also effective in stimulating DA synthesis and release, thus dietary phenylalanine supplementation can increase DA biosynthesis in fish.

Item does not contain fulltext Senegalese sole (Solea senegalensis) is a eurytherm teleost that under natural conditions can be exposed to annual water temperature fluctuations between 12 and 26 degrees C. This study assessed the effects of temperature on sole metabolic status, in particular in what concerns plasma free amino acid changes during thermal acclimation. Senegalese sole maintained at 18 degrees C were acclimated to either cold (12 degrees C) or warm (26 degrees C) environmental temperatures for 21 days. Fish maintained at 18 degrees C served as control. Plasma concentrations of cortisol, glucose, lactate, triglycerides, proteins, and free amino acids were assessed. Cold acclimation influenced interrenal responses of sole by increasing cortisol release. Moreover, plasma glucose and lactate concentrations increased linearly with temperature, presumably reflecting a higher metabolic activity of sole acclimated to 26 degrees C. Acclimation temperature affected more drastically plasma concentrations of dispensable than that of indispensable amino acids, and different acclimation temperatures induced different responses. Asparagine, glutamine and ornithine seem to be of particular importance for ammonia detoxification mechanisms, synthesis of triglycerides that may be used during homeoviscous adaptation and, to a lesser extent, as energetic substrates in specimens acclimated to 12 degrees C. When sole is acclimated to 26 degrees C taurine, glutamate, GABA and glycine increased, which may suggest important roles as antioxidant defences, in osmoregulatory processes and/or for energetic purposes at this thermal regimen. In conclusion, acclimation to different environmental temperatures induces several metabolic changes in Senegalese sole, suggesting that amino acids may be important for thermal acclimation. 01 juli 2012

The effects of prolonged feed deprivation were assessed on blood plasma metabolites and free amino acid levels in Solea senegalensis. Juvenile specimens were maintained at two experimental conditions (24 h fasted and 21 days feed-deprived). In feed-deprived fish, relative growth rate and hepatosomatic index as well as plasma triglycerides and proteins levels were significantly lower. However, plasma cortisol levels were higher in feed-deprived fish, while plasma glucose and lactate values were not significantly different between treatments. Furthermore, feed-deprived fish showed higher levels of total plasma free amino acids than 24 h fasted fish. In 21 days feed-deprived sole, an increase in plasma cortisol levels may suggest a functional role in mobilizing energy due to the catabolic action of this hormone in teleosts. Higher levels of glutamine, arginine and ornithine in 21 days feed-deprived fish may be indicative of a dual role for these amino acids: ammonia detoxification and carbon source for gluconeogenesis. The increased plasma glucogenic and branched-chain amino acid levels, together with the maintenance of plasma glucose in 21 days feed-deprived sole, suggest active liver gluconeogenic processes supported by tissue proteolysis.

Fish held at high stocking densities are generally exposed to chronic stress situations that impose severe energy demands and may predispose the fish to infection. Senegalese sole Solea senegalensis (Kaup) juveniles (78.8±18.9 g body weight) were maintained at low (LSD; 4 kg m−2 at the end of the experiment), medium (MSD; 9 kg m−2) and high (HSD; 14 kg m−2) stocking densities during a period of 63 days. Although disease outbreaks were observed in fish reared at HSD, growth and food consumption did not vary among different treatments. Results from plasma cortisol and free amino acids (FAAs) showed significant differences among different rearing densities pointing to HSD as stressful rearing condition. However, higher plasma glucose and osmolality levels indicated that fish held at MSD may also be under stress. The higher usage of FAAs from the HSD group may be due to the higher demand for energy production in order to cope with stressful rearing conditions, higher rate of protein synthesis or due to synthesis of other important metabolites related to stress response. Therefore, results from the present study point out to HSD as a stressful rearing condition and suggest that crowding stress may affect amino acid requirements.

The objective of this work was to evaluate the effect of a dietary amino acid imbalance, originating from the use of a soy protein concentrate (SPC) as the major protein source, on the growth performance and amino acid metabolism of Senegalese sole (Solea senegalensis) postlarvae. Senegalese sole (85.6±24.6 mg wet weight) were fed one of two experimental diets: one based on fish meal (FM) and another based on SPC. Diets were isonitrogenous (around 56% crude protein) and isoenergetic. Diet acceptability was very good and the growth rate was 6.9% day−1 for sole eating the FM diet and 6.0% day−1 for sole eating the SPC diet. Mass-specific ammonia excretion and the activities of selected amino acid metabolic enzymes (ALAT, ASAT and GDH) did not present significant differences between treatments, although this may have been due to the high variability found for these parameters in the SPC treatment. This variability may suggest different capacities of individual fish to adapt to the possible methionine dietary deficiency. The utilization of amino acids as a substrate for lipogenesis does not seem to be affected by the dietary protein source, since NAPDH-generating enzymes (G6PD and ME) had similar activities in both treatments. Amino acid metabolism in Senegalese sole postlarvae seems to be slightly affected by the dietary protein source. Nevertheless, the changes induced by the SPC diet do not seem to impair growth, at least at the high dietary protein level used in this experiment.

The postprandial metabolism of dietary free amino acids (AA) was studied in post-larval Senegal sole, Solea senegalensis, aged 32 days after hatching (DAH). The diet was administered as a single pulse (36 nl, 43.1 mmol/l) using a micro tube-feeding technique and a dissolved mixture of crystalline AA. In four separate treatments the diet contained L [U-(14)C] tracer for two indispensable AA (IAA), lysine and arginine or two dispensable amino acids (DAA), glutamate and alanine. The post-larva absorbed all tested AA from the diet with similar efficiency (97.5%). A small fraction of the IAA was catabolised (11.5+/-1.1 and 15.1+/-3.3%, for lysine and arginine, respectively) and a high proportion was retained in the body (86.7+/-1.3 and 81.6+/-4.1%). For the DAA more were catabolised (64.9+/-5.3% and 41.4+/-7.2% for glutamate and alanine, respectively) and less were retained (32.9+/-5.1% and 56.3+/-7.2%). On this basis, it appears that post-larval Senegal sole use DAA in preference to IAA as energy substrates while the retention (assimilation efficiency) is better for the IAA. These results support other recent studies that early stages of fish have a better capacity to regulate AA catabolism than previously believed and that indispensable AA are saved for body growth.

This study evaluated the effect of dietary taurine supplementation on the growth performance and methionine metabolism of gilthead seabream larvae. For this purpose, a growth experiment was carried out in which seabream larvae were fed upon rotifers supplemented with “blank” or taurine-enriched liposomes. A complementary trial was also done in which seabream larvae fed with live prey were subsequently tube-fed a solution containing a L-[U-14C] methionine with or without a taurine supplement. Results from the growth experiment showed that rotifers were successfully enriched with taurine, but no effects were observed on larval growth performance, survival or amino acid composition. Furthermore, dietary taurine supplementation did not result in an increase of larval taurine levels, a factor that may have been determinant for the absence of effects observed on growth performance. In the tube-feeding trial, results showed that dietary taurine supplementation led to an increase of methionine retention in larvae. These findings suggest the existence of an active taurine biosynthesis pathway for gilthead seabream during the larval stage. Hence, gilthead seabream may not be dependent on dietary taurine to maintain the taurine body pool, since it may convert taurine from methionine if required. Taken together, the results from this study indicate that dietary taurine supplementation does not seem to enhance the larval growth performance in fish species able to biosynthesise taurine during this stage, which seems to be the case of gilthead seabream. However, this study also showed that in these species, dietary taurine supplementation may ultimately affect larval metabolism by increasing methionine availability for several important physiological purposes, contributing to a better understanding on the role of taurine during the early life stages of fish development.

Flatfish species seem to require dietary taurine for normal growth and development. Although dietary taurine supplementation has been recommended for flatfish, little is known about the mechanisms of taurine absorption in the digestive tract of flatfish throughout ontogeny. This study described the cloning and ontogenetic expression of the taurine transporter (TauT) in the flatfish Senegalese sole (Solea senegalensis). Results showed a high similarity between TauT in Senegalese sole and other vertebrates, but a change in TauT amino acid sequences indicates that taurine transport may differ between mammals and fish, reptiles or birds. Moreover, results showed that Senegalese sole metamorphosis is an important developmental trigger to promote taurine transport in larvae, especially in muscle tissues, which may be important for larval growth. Results also indicated that the capacity to uptake dietary taurine in the digestive tract is already established in larvae at the onset of metamorphosis. In Senegalese sole juveniles, TauT expression was highest in brain, heart and eye. These are organs where taurine is usually found in high concentrations and is believed to play important biological roles. In the digestive tract of juveniles, TauT was more expressed in stomach and hindgut, indicating that dietary taurine is quickly absorbed when digestion begins and taurine endogenously used for bile salt conjugation may be recycled at the posterior end of the digestive tract. Therefore, these results suggest an enterohepatic recycling pathway for taurine in Senegalese sole, a process that may be important for maintenance of the taurine body levels in flatfish species. Electronic supplementary material The online version of this article (doi:10.1007/s00726-010-0826-6) contains supplementary material, which is available to authorized users.

Submitted by Cláudia Aragão (caragao@ualg.pt) on 2014-06-27T15:54:39Z No. of bitstreams: 1 Changes in plasma amino acid levels in a euryhaline fish exposed to different environmental salinities.pdf: 266884 bytes, checksum: 641d084627ff64add627207a0fa103fa (MD5) Approved for entry into archive by Carla Janeira (cjaneira@ualg.pt) on 2014-07-02T13:41:46Z (GMT) No. of bitstreams: 2 0467513760195905.zip: 232033 bytes, checksum: 965c98a95d931054253620cfc3e8a833 (MD5) Changes in plasma amino acid levels in a euryhaline fish exposed to different environmental salinities.pdf: 266884 bytes, checksum: 641d084627ff64add627207a0fa103fa (MD5) Made available in DSpace on 2014-07-02T13:41:46Z (GMT). No. of bitstreams: 2 0467513760195905.zip: 232033 bytes, checksum: 965c98a95d931054253620cfc3e8a833 (MD5) Changes in plasma amino acid levels in a euryhaline fish exposed to different environmental salinities.pdf: 266884 bytes, checksum: 641d084627ff64add627207a0fa103fa (MD5) Previous issue date: 2010

Aromatic amino acids (AAs, phenylalanine and tyrosine) may be specifically required during fish metamorphosis, since they are the precursors of thyroid hormones which regulate this process. This project attempted to evaluate aromatic AA metabolism during the ontogenesis of fish species with a marked (Senegalese sole; Solea senegalensis) and a less accentuated metamorphosis (gilthead seabream; Sparus aurata). Fish were tube-fed with three L-[U-14C] AA solutions at pre-metamorphic, metamorphic and post-metamorphic stages of development: controlled AA mixture (Mix), phenylalanine (Phe) and tyrosine (Tyr). Results showed a preferential aromatic AA retention during the metamorphosis of Senegalese sole, rather than in gilthead seabream. Senegalese sole's highly accentuated metamorphosis seems to increase aromatic AA physiological requirements, possibly for thyroid hormone production. Thus, Senegalese sole seems to be especially susceptible to dietary aromatic AA deficiencies during the metamorphosis period, and these findings may be important for physiologists, fish nutritionists and the flatfish aquaculture industry.

Submitted by Cláudia Aragão (caragao@ualg.pt) on 2014-07-03T09:35:03Z No. of bitstreams: 1 Stress response and changes in amino acid requirements in Senegalese sole (Solea senegalensis Kaup 1858).pdf: 188488 bytes, checksum: 14dd8d76a6c88ef43ece205db64f4d3e (MD5) Approved for entry into archive by Merja Muzavor (mmuzavor@ualg.pt) on 2014-07-04T13:00:39Z (GMT) No. of bitstreams: 2 0467513760195905.zip: 145125 bytes, checksum: 4e32b8c6a1f11283065eb97b02d855ec (MD5) Stress response and changes in amino acid requirements in Senegalese sole (Solea senegalensis Kaup 1858).pdf: 188488 bytes, checksum: 14dd8d76a6c88ef43ece205db64f4d3e (MD5) Made available in DSpace on 2014-07-04T13:00:39Z (GMT). No. of bitstreams: 2 0467513760195905.zip: 145125 bytes, checksum: 4e32b8c6a1f11283065eb97b02d855ec (MD5) Stress response and changes in amino acid requirements in Senegalese sole (Solea senegalensis Kaup 1858).pdf: 188488 bytes, checksum: 14dd8d76a6c88ef43ece205db64f4d3e (MD5) Previous issue date: 2008

7 páginas, 2 figuras, 4 tablas., [EN] Live preys commonly used in fish larval rearing seem to be imbalanced in terms of amino acids. Manipulation of their amino acid composition is difficult, but the use of microencapsulated diets allows this manipulation. This study analysed the effect of amino acid supplementation, in order to compensate for dietary amino acid imbalances, on growth and survival of gilthead seabream (Sparus aurata) larvae. Larvae were reared until 32 days after hatching, in a closed recirculating water system (19 °C), using only live food (rotifers and Artemia). Thereafter, larvae were fed Artemia or one of the experimental microencapsulated diets: supplemented with indispensable amino acids (IAAsup diet), or supplemented with dispensable amino acids (DAAsup diet). Experiment lasted for 10 days. Dietary supplementation with indispensable amino acids resulted in A/E ratios [(each indispensable amino acid) × (total indispensable amino acids)−1 × 1000] more similar to the ones of larval seabream and in higher IAA:DAA ratios than in the DAAsup diet. Survival was similar in larvae fed the IAAsup diet (75%) or Artemia (87%), but was significantly lower in larvae fed the DAAsup diet (52%). Larvae from all treatments more than doubled their average dry weight during the experimental period. Final dry weight was similar in larvae fed both microcapsules, but these were lighter than larvae fed with Artemia. Relative growth rate (RGR) and total biomass production tended to be higher in larvae fed the IAAsup (RGR = 9% day−1) than the DAAsup diet (RGR = 7.5% day−1) and only in this last treatment these parameters were significantly lower than in larvae fed with Artemia. Therefore, dietary supplementation with indispensable amino acids resulted in a more balanced dietary amino acid profile, which significantly increased survival. Further studies introducing microdiets earlier in the development seem necessary in order to optimise growth., [FR] Les proies vivantes utilisées dans les élevages larvaires semblent déséquilibrées en terme d'acides aminés. Manipuler la composition en acides aminés est difficile mais l'usage d'aliments encapsulés le permet. Cette étude analyse l'effet d'une supplémentation en acides aminés sur la croissance et la survie des larves de daurade royale (Sparus aurata) afin de compenser un déséquilibre. Les larves sont élevées jusqu'à 32 jours après éclosion en circuit fermé (eau à 19 °C), en utilisant des aliments vivants uniquement (rotifères et Artemia). Les larves sont ensuite nourries avec des Artemia ou bien avec des aliments microencapsulés expérimentaux, supplémentés en acides aminés soit indispensables (régime IAAsup) soit non-indispensables (régime DAAsup). La durée de l'expérience est de 10 jours. La supplémentation en acides aminés indispensables conduit à des ratios A/E [(chaque AA indispensable) × (total en AA indispensables)−1 × 1000] plus proches de ceux de la larve de dorade et des ratios IAA/DHA plus élevés que ne le fait la supplémentation en acides aminés non-indispensables. Le taux de survie est similaire chez les larves nourries avec le régime IAAsup (75 %) ou Artemia (87 %) mais inférieur significativement chez les larves soumises au régime DAAsup (52 %). Le poids sec moyen de ces larves, quel que soit leur régime alimentaire, a doublé durant la période expérimentale. Le poids sec est similaire chez les larves nourries de microcapsules mais moins élévé chez celles nourries d'Artemia. Le taux de croissance relative (RGR) et la production de biomasse totale tendent à être plus élevés chez les larves nourries en IAAsup (RGR = 9 % jour−1) que celles nourries en DAAsup (RGR = 7,5 % jour−1). C'est seulement dans ce dernier traitement que les paramètres sont plus faibles significativement que chez les larves nourries d'Artemia. Ainsi, les supplémentations alimentaires en acides aminés indispensables peuvent augmenter la survie. D'autres études, introduisant des micro-aliments de façon plus précoce dans le développement, semblent nécessaires afin d'optimiser la croissance., The authors acknowledge funding by project AGL2004-06669-C02-01 (MCYT, Spain) and grant SFRH/BPD/14835/2003 (FCT, Portugal).

The rearing of most marine fish larvae still relies on live food. Dietary amino acid (AA) imbalances when using live food in the larval rearing of flatfishes have been suggested. The aim of this study was to test if dietary AA supplementation affects AA metabolism in Senegalese sole (Solea senegalensis) post-larvae. This was done by tube-feeding Artemia-fed sole with a dipeptide solution containing two potential limiting AA (leucine and phenylalanine), in order to supplement the larval gut content and to balance the dietary AA profile. Two experiments were done using different 14C-labelled AA as tracers. The first used a 14C-protein hydrolysate to test the effect of balancing the dietary AA profile on the overall AA metabolism. The second experiment analysed the effect of balancing the dietary AA profile on metabolism of three-selected AA: arginine, leucine and glutamate. A set-up to determine the handling of 14C-labelled AA was used, in order to quantify the absorption, catabolism and retention of the test mixtures by the sole post-larvae. The first experiment demonstrated that, when fish were fed the dipeptide supplement, AA catabolism decreased and AA retention increased. This agrees with the hypothesis that balancing the dietary AA profile increases AA retention in fish. In the second experiment, arginine and glutamate catabolism were reduced by the dipeptide supplementation. Leucine supplementation did not reduce leucine catabolism but instead increased arginine and glutamate retention. This result supports earlier studies that fish larvae are able to regulate their AA metabolism. A balanced dietary AA profile increases the AA retention and may improve growth and nitrogen utilisation.

Texto integral disponível através do editor To improve the formulation of diets for the early stages of marine fish, assimilation rates of free amino acids (FAA) and protein in postlarval Senegal sole (Solea senegalensis) were determined. Fish (2.45 ± 0.87 mg dry weight) were tube fed 36 nL of a diet of FAA containing L-[35S] methionine (FAA diet) or bovine serum albumin, containing L-[methylated-14C]bovine serum albumin (Prot-diet), both at a concentration of 4.08 g/L. A time series was performed, and the amounts of label in incubation water, liver, gut and body carcass were quantified. The FAA diet was absorbed with a 3.5-times-higher transfer rate (P < 0.001) from the gut into the larval body tissues compared with the Prot-diet. The FAA diet also was assimilated with greater efficiency than the Prot-diet (80% versus 58%, P = 0.001). If we assume that the label present in the gut represents amino acids incorporated into the intestinal tissue, the assimilation efficiencies for the two diets were 89 and 64%. Therefore, FAA seems to be superior to protein as a dietary source of amino acids in Senegal sole postlarvae. However, because the absorption dynamics of protein and FAA differ, care should be taken when using the sources together to avoid amino acid imbalance.