Utilization of free amino acids, yolk proteins and lipids in developing eggs and yolk-sac larvae of walleye pollock Theragra chalcogramma

ABSTRACT:   To elucidate the utilization of the major yolk nutrient stocks in eggs and larvae of walleye pollock Theragra chalcogramma , the contents of free amino acids (FAA), the major yolk protein (180 kDa lipovitellin originated from vitellogenin B in ovulated eggs: oLv B), and lipids were measured. Most eggs hatched 18 days after fertilization at 5°C, and all larvae absorbed almost all their yolk mass by 28 days. The total FAA content showed no change during the first 6 days, and then decreased to 28% of the initial level by 18 days. The oLv B contents, measured by an enzyme-linked immunosorbent assay using a specific antiserum against oLv B, gradually decreased from 6 to 18 days, followed by a rapid decline. The content of phospholipids (PL) and triacylglycerols (TG) showed no marked change until hatching, and then decreased until disappearance of yolk sac. From these results, it is proposed that there are two main periods for nutrient utilization in embryos and larvae of walleye pollock. In the first period, FAA was mainly utilized until 18 days after fertilization. Active utilization of oLv B and lipids (PL and TG) instead of FAA occurred during the second period from 18 to 28 days.     

海水仔稚鱼早期阶段氨基酸的营养生理研究进展

深海海鱼鱼卵中的游离氨基酸占鱼卵总氨基酸含量的近50％（干重），游离氨基酸库中的游离氨基酸似乎是卵黄蛋白的水解产物。仔鱼吸收卵黄内源营养的过程中，游离氨基酸库中的游离氨基酸逐渐减少，到仔鱼开始摄食时，库中的游离氨基酸已近枯竭，游离氨基酸不仅是代谢能源，而且是体蛋白合成的基本原料，仔鱼开始摄食后，氨基酸作为分解代谢的重要物质，可能有高达60％的能量由氨基酸提供，由于仔稚鱼的生长主要是通过合成蛋白质增加鱼体的重量，加之仔稚鱼的生长速度很高，因此，仔稚鱼需要其饲料中有较高的氨基酸含量。仔鱼开始摄食后，其消化系统对蛋白质的水解和吸收能力较弱，海水仔稚鱼的早期阶段，仔鱼的肠道对游离氨基酸的吸收要高于对多肽和蛋白质中氨基酸的吸收，仔鱼开始摄食后，由浮游生物获得大量的游离氨基酸，用微粒饲料培育仔稚鱼，微脂粒技术可能是向饲料中添加游离氨基酸的实用技术。     

Free Amino Acids in Pacu,Piaractus mesopotamicus,Eggs and Larvae

Free amino acids (FAA) are used principally as substrate in protein synthesis and the source of energy in aerobic catabolism. In marine fish, embryo and larvae FAA are used to maintain body fluid osmolality during fish early development. However, there is essentially no information about FAA concentrations in early ontogeny of freshwater neotropical species in comparison to marine fishes. Therefore, the aim of this study was to evaluate the FAA concentrations in pacu, Piaractus mesopotamicus, eggs and larvae. Broodstock fish were induced to spawn and ovulated females were stripped of their eggs and immediately sampled for analysis. Larvae were sampled right after hatching (HL) and after the completion of the yolk‐sac absorption (YSA). The wet weight of the HL and YSA larvae amounted to 0.5 ± 0.1 mg and 1.1 ± 0.3 mg, respectively. HL larvae showed higher levels of most of the indispensable amino acids (IAA) in comparison to eggs and YSA larvae. Exceptions were observed with His and Trp that showed higher or similar levels, respectively, in YSA larvae. The FAA Orn, Tau, Glu, Gln, Gly, and Tyr increased concentrations in both larval stages while that of Tau was found in higher concentration in all analyzed stages. Also, the concentrations of Asn, Ala, Pro, Ser, and Asp were higher in HL larvae. Both larval stages displayed a rise in total free IAA/total free DAA (dispensable amino acids) ratio. The authors conclude that the highest level of FAA in HL pacu larvae is indicative of active proteolysis of yolk reserves and a probable catabolism regulation of some FAA through spare‐effect. In addition, Tau is one of the major FAA occurring during pacu ontogeny and may be performing regulation on body fluid osmolality regulation .     

Chemical changes in fed and starved larval trout cod, Maccullochella macquarensis during early development

The changes in proximate composition, amino acid (total and free) and fatty acid content of artificially propagated trout cod, Maccullochella macquariensis larvae from five mothers hatched, weaned and reared separately, each in two groups, one fed with Artemia naupli and the other starved, for 15 days (after yolk resorption), are presented. There was no significant change in the proximate composition of fed larvae with devlopment, but in starved larvae the protein (linearly) and lipid (curvi-linearly) content decreased significantly as starvation progressed. The essential amino acids (EAA) and non- essential amino acids (NEAA) found in highest amounts in trout cod larvae were lysine, leucine, threonine and arginine, and alanine, serine and glutamic acid, respectively. In fed larvae the total amino acid (TAA), TEAA and TNEAA content did not vary significantly as development progressed. In starved larvae the TAA, EAA and NEAA content, as well as all the individual amino acids decreased significantly (P<0.05) from the levels in day of hatch and/or yolk-sac resorbed larvae. The greatest decrease occurred in the TEAA content (7.38±0.76 at day of hatch to 1.96±0.09 15 day starved in moles larva^–1; approximately a 74% decrease), whereas the decrease in TNEAA was about 38%. Unlike in the case of TAA distinct changes in the free amino acid (FAA) pool were discernible, from day of hatch and onwards, in both fed and starved trout cod larvae. In both groups of larvae the most noticeable being the decrease of % FEAA in TFAA, but not the % FAA in TAA. Four fatty acids together, accounted for more than 50% of the total in each of the major fatty acid categories in all larvae sampled; 16: 0, 18:1n-9, 22: 6n-3 and 20: 4n-6, amongst saturates, monoenes, n-3 PUFA and n-6 PUFA, respectively. Twelve fatty acids either decreased (14: 0, 16: 1n-7, 20: 1n-9, 20: 4n-6, 20: 5n-3, 22: 5n-3 and 22: 6n-3) or increased (18: 2n-6, 18: 3n-3, 18: 3n-6, 18: 4n-3 and 20: 3n-3) in quantity, after 15 days of feeding, from the base level in day of hatch and/ or yolk- sac resorbed larvae. The greatest increase occurred in 18: 3n-3 from 6.4±0.1 to 106.2±13.1 g mg lipid^–1 larva^–1, and the greatest decrease occurred in 22: 6n-3 (181.2±12.4 to 81.4±6.2 g mg lipid^–1 larva^–1). In starved larvae, at the end of 15 days, all the fatty acids, except 18: 0, 20: 3n-3 and 20: 4n-6, decreased significantly (P<0.05) from the levels in day of hatch and/or yolk- sac resorbed larvae.     

Changes in free amino acid profile of red snapper <Emphasis Type="Italic">Lutjanus campechanus</Emphasis>, eggs,and developing larvae

The free amino acids (FAA) profile was determined for newly fertilized eggs and resultant larvae from wild-caught red snapper Lutjanus campechanus induced to spawn with hCG. Yolk sac and oil globule volumes of eggs and larvae were monitored over time from digital photographs. FAA profiles of the eggs and larvae were measured in picomoles (pmol) of FAA/mg of eggs by HPLC. Newly fertilized eggs had a mean total FAA content of 21.72 ± 3.55 nmoles/egg (92.81 ± 9.71 nmoles/mg eggs). Leucine, valine, lysine, and isoleucine were the most abundant essential FAA comprising 35.9% of the total FAA. Alanine, serine, asparagine, and glycine were the most abundant non-essential FAA comprising 34.2% of the total FAA. At 24 h post-hatch (hph) the mean total FAA had decreased by 81% since egg fertilization. The bulk of the FAA decrease was between the time of hatch and 12 hph. Only 8.5 ± 1.5% of the initial concentration in fertilized eggs of isoleucine, 9.7 ± 2.5% of arginine, and 9.9 ± 2.0% of threonine remained at 12 hph. Among the non-essential FAA, alanine dropped the most by 12 hph with 4.6% of the concentration found in a recently fertilized egg remaining, while cysteine had increased 254.7 ± 26.2%. The yolk sac volume decreased rapidly in the first 12 hph and was further reduced 77.0 ± 2.5% from 12 to 24 hph. The oil globule depletion rate was a more linear decline from fertilized egg to 36 hph.     

Amino acid profiles and amino acid utilization in larval African catfish (Clarias gariepinus): effects of ontogeny and temperature

The present study investigated the qualitative amino acid (AA) requirements of larval African catfish Clarias gariepinus. Yolk-sac larval AA profiles were measured at different temperatures and also in animals reared at 28 °C fed Artemia nauplii or an experimental dry diet. The AA profile of C. gariepinus larvae changed during ontogeny, especially before the start of exogenous feeding. The AA profiles of the food items (yolk, Artemia and the dry diet) differed considerably from that of the larvae. No selective absorption of yolk AA was detected. Higher temperatures led to increased absorption and depletion rates of AA, and also to a higher retention efficiency of yolk nutrients. However, changes in temperature did not induce preferential absorption or depletion of individual AA, and caused only small variations in the AA profile. Depletion rates of individual AAs varied, possibly due to differences between larval and yolk AA profiles, and also to changes in the larval AA profile during ontogeny. There was little regulation of catabolism of individual AA in yolk-sac and starved larvae, and no sparing of essential AA.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

The amino acid profiles in developing eggs and larvae of the freshwater Percichthyid fishes,trout cod,Maccullochella macquariensis and Murray cod,Maccullochella peelii peelii

Results on changes in the total amino acids (protein bound + free) and the free amino acids (FAA) in relation to development, from egg (unfertilised and/or fertilised) to yolk-sac resorbed larva, before first feeding, in two Percichthyid fish, trout cod, Maccullochella macquariensis and Murray cod, Maccullochella peelii peelii, which lay demersal, adhesive eggs, are presented. Throughout development, the FAA accounted for only a small proportion (0.19 % in fertilised eggs of both species) of the total amino acid pool. Nine essential amino acids (EAA) and eight non-essential amino acids (NEAA) were quantified in the amino acid pool at all stages of development. In both species, the total amino acid content decreased during the transformation (at 20 ± 1 °C) from newly hatched larva to yolk-sac resorbed larva. Overall, the changes in the TEAA and TNEAA reflected that of the amino acid pool. In trout cod, all but one EAA (lysine) and two NEAA (cysteine and glycine) decreased with ontogeny, from fertilised egg to yolk-sac resorbed larva. In Murray cod, however, the exceptions to the general decline were two NEAA (aspartic acid and glycine). In contrast, the FAA increased with development, the changes being reflected in both FEAA and FNEAA. Qualitatively, the predominant free amino acids in trout cod and Murray cod eggs were alanine, lysine, leucine and serine. Because the egg protein and the total amino acid contents declined with development, it is concluded that the rate of breakdown of yolk protein was higher than the anabolic and catabolic processes during embryogenesis. Data also suggest that in freshwater fish FAA are an unlikely primary energy substrate during embryogenesis.     

Free amino acid and protein content in the planktonic copepod Temora longicornis compared to Artemia franciscana

Previous studies have indicated that natural zooplankton, the natural prey organisms of marine fish larvae, is superior to enriched Artemia spp. nauplii in supporting survival, growth and normal development of pigmentation and eye migration in Atlantic halibut larvae. The present study was designed to compare the amino acid dynamics in copepods harvested from a natural lagoon with that of enriched Artemia franciscana nauplii. The natural zooplankton in the present study was dominated by Temora longicornis, but varied in its developmental stage composition, apparent nutritional quality, amount and availability. The protein content in natural zooplankton varied from 31% to 54% of dry mass (DM) compared to 31% in enriched Artemia nauplii. The amount of free amino acids (FAA) in relation to protein was 14% in enriched Artemia nauplii and varied between 16% and 27% in zooplankton in two consecutive seasons. The FAA composition of zooplankton in 1996 was unaffected by stage and season, and showed a species-specific pattern. In response to starvation, two patterns in the amino acid dynamics of copepods were found. The first, exhibited by the major nonessential amino acids (NEAA), showed a continuous reduction in individual amount, while the second pattern, exhibited by the minor nonessential amino acids and all the essential FAA, had an initial increase followed by a reduction. It is suggested that the protein amount in DHA Selco™ (INVE)-enriched Artemia nauplii is too low in order to maximise the growth potential of some marine fish larvae as cod and halibut.     

The impact of varying feeding regimes on oxygen consumption and excretion of carbon dioxide and nitrogen in post-smolt Atlantic salmon Salmo salar L.

Oxygen consumption, carbon dioxide excretion and nitrogen excretion from 2 kg Atlantic salmon Salmo salar L. were studied at land-based outdoor tanks, throughout a 10 day period. Fish were fed six feed rations (0 [fasting fish], 0.15, 0.30, 0.45, 0.60 and 0.75% BW day^-1) of commercially extruded dry feed at two provision regimes: between 07.00-09.00 h and 19.00–21.00 h (periodic feeding); and between 07.00-21.00 h (continuous feeding). Fish were acclimatized to the feeding regime for 5 days prior to the start of the experiment. From days 5 to 10 of the study, oxygen consumption was measured automatically every 15 min. During day 10, carbon dioxide excretion, ammonia and urea excretion were measured hourly throughout a 24 h period. During the experiment, the water temperature and salinity were 8.5C and 33.5 ppt, respectively. Significant linear relationships between feed ration and metabolic rates were evident. Increased feed ration influenced oxygen consumption, carbon dioxide excretion, ammonia and urea excretion, ammonia quotient and the respiration quotient, proportionally. The two feed provision regimes caused the establishment of different daily rhythms in metabolic excretion. Only small differences in total daily excretion were however recorded for each feed ration. Ammonia and respiratory quotient results clearly showed that fish became more dependent on fat oxidation as an energy source, when feed was restricted.     

Free amino acids in metamorphosing bonefish (Albula sp.) leptocephali

Metamorphosing leptocephalous larvae of the bonefish (Albula sp.) were analyzed for total ninhydrinpositive substances (NPS) and free amino acids. Total NPS content showed little change during metamorphosis. The average NPS value (±S.E.) for 16 larvae was 1.8 (±0.1) mg×larva^–1, which represents approximately 4% and 8% of the total dry weight of early and advanced larvae, respectively. Taurine was the most abundant free amino acid in whole-larva extracts, accounting for 36% and 59% of the total by weight in early and advanced larvae, respectively. The essential amino acids, leucine, isoleucine, phenylalanine, histidine, valine, methionine, lysine and arginine, accounted for about half (47%) of the total in early larvae but were reduced to about 23% of the total in advanced larvae. All of the component essential amino acids decreased during metamorphosis, but the greatest effect was seen with the first five. The remaining non-essential amino acids comprised less than 20% of the total in early larvae and, although the overall value changed little during metamorphosis, certain components such as glycine and glutamic acid showed large increases whereas others such as tyrosine and serine were reduced. Increases in amino acid content after acid hydrolysis of whole-larva extracts indicated that trichloroacetic acid-soluble, low molecular weight peptides were present in both early and advanced leptocephali.     

Kinetics of Nitrogen Excretion by Cultured Flounder Paralichthys olivaceus

This study was undertaken to quantify the rate of nitrogen excretion by the cultured Japanese flounder as a function of body weight and water temperature. Measurements of the did rate of nitrogen excretion for the cultured flounder were made at mean water temperature of about 16, 19, and 23 C. The excretion rate of nilmgen was estimated by adding the nitrogen excreted in the forms of ammonia and urea nitrogens. The diurnal pattern in the excretion rate of nitrogen was governed mainly by the feeding time. The proportions of excreted ammonia to total nitrogen were 77.9% at 16 C, 84.5% at 19 C, and 83.2% at 23 C. The average was 81.9%. The relationship between the body weight and excretion rate of nitrogen ( E_N , mg N/fish per h) is expressed as E_N =α,.W', where α, = nitrogen excretion rate at temperature 0 . The parameter α, is dependent on water temperature, such that α, =α₂₀ Q₀₁^(0–200) where α₂₀= nitrogen excretion rate at 20 C, and Q₀₁, = temperature coefficient. The values of the kinetics parameters for the cultured flounder were found to be γ = 0.57, α₂₀= 0.060, and Q₀₁= 1.12.     

Growth, stress response and free amino acid levels in Senegalese sole (Solea senegalensis Kaup 1858) chronically exposed to exogenous ammonia

Stressful husbandry conditions are likely to affect growth and amino acid metabolism in fish. In this study, chronic ammonia exposure was used to test the effects of a stressor on growth and amino acid metabolism of Senegalese sole juveniles. The fish were exposed for 52 days to 11.6 mg L⁻¹ [low‐TAN (L‐TAN)] or 23.2 mg L⁻¹ [high‐TAN (H‐TAN)] of total ammonia nitrogen (TAN), or to 0 mg L⁻¹ (Control). Growth in L‐TAN groups was slightly but significantly different from the Control groups [relative growth rate (RGR=0.35±0.13 and 0.52±0.23% day⁻¹ respectively)]. In H‐TAN groups, growth was severely affected (RGR=0.01±0.13% day⁻¹). Stress parameters (plasma cortisol and glucose) showed slight or no significant differences between treatments. Plasma free amino acid (FAA) concentrations were affected after H‐TAN treatment. Increases in glutamine and aspartate concentrations in H‐TAN fish suggest alterations in amino acid metabolism related to nitrogen excretion processes. Some of the changes in FAA concentrations also suggest mobilization to energy supply and synthesis of metabolites related to stress response. Therefore, Senegalese sole seem to adapt to the L‐TAN concentration tested, but the H‐TAN concentration reduced growth and affected amino acid metabolism.     

Absorption, assimilation and catabolism of individual free amino acids by larval Atlantic halibut (Hippoglossus hippoglossus)

Following the completion of the lecithotrophic phase, most marine larvae rely on an incompletely developed digestive tract to absorb amino acids (AA) and other nutrients needed for rapid growth. Despite their undeveloped state, larvae must absorb nutrients in sufficient amounts to fuel exceptionally high rates of growth. This study examined the ability of larval Atlantic halibut to absorb, assimilate and catabolise dispensable (alanine, glutamate) and indispensable (arginine, lysine) dietary free amino acids (FAA) using tube-fed AA solutions with ¹⁴C tracers. Absorption of FAA was rapid with an average of 71% absorbed from the gut within 30 min after tube feeding. Evacuation of FAA by larvae was low, averaging only 6% of tube-fed dose. Dispensable amino acids (DAA) were catabolised in greater proportion (17%) than indispensable FAA (54%). Saturation of FAA transporters was not attained under the present conditions (20 mM, 20% midgut filling). Absorption rates did not differ significantly between the four FAA.     

Incorporation of yolk fatty acids into body lipids of goldfish (Carassius auratus L.) larvae raised at two different temperatures

In five separate experiments, eggs from a single female goldfish were fertilized at 20°C. They were incubated at 22°C for 6 hours, after which some of the eggs were transferred to 13°C. When a defined post-hatch developmental stage was reached, lipid extracts were prepared from larvae, both with yolk sacs intact and after removal of the yolk sac by dissection. Other larvae were sampled at yolk exhaustion. Gas chromatographic analysis of fatty acid profiles revealed that larvae incorporated 16:0, 18:0, 20:4 (n–6) and 22:6 (n–3) into their tissues in proportions higher than those present in the eggs from which they were derived. At 22°C, these trends were particularly apparent at yolk exhaustion. At 13°C, proportions of polyunsaturated fatty acids in the bodies of newly hatched larvae were higher than those in the 22°C larval bodies. Monounsaturated fatty acids were preferentially depleted during development, especially in larvae from high quality eggs. No dependence of egg quality, as assessed by larval viability at 22°C, on total egg lipid mass or fatty acid composition was found. Larvae from the lowest quality eggs showed a reduced preference for incorporation of (n–3) polyunsaturated fatty acids into their tissues.     

Augmentation of free amino acids in eggs of red snapper,Lutjanus campechanus as part of the induced spawning protocol

An attempt was made to increase free amino acids (FAA) concentrations in eggs of red snapper Lutjanus campechanus as part of an induced spawning protocol. Mature female red snapper were given intramuscular injections of human chorionic gonadotropin (HCG) at a 1100 IU kg^?1 of female body weight. Immediately after the HCG injection, one set of females received an intramuscular injection of FAA in a buffered saline solution at a dose of 20 mg FAA cocktail kg^?1 of body weight. The FAA cocktail was 25% isoleucine, 25% leucine, 25% lysine and 25% valine on a molecular weight basis. A second set of fish did not receive the FAA supplement. Both sets of fish were equally successful in spawning with ovulation occurring 21–32.5 h post injection. Injection of FAA into brooders as eggs began final maturation and hydration resulted in an increase in leucine (nmoles mg egg^?1) and a slight increase in isoleucine (nmoles egg^?1) in recently ovulated eggs. FAA supplementation via brood injection altered FAA utilization rates by embryos and during larval development, resulted in a greater yolk sac and oil globule reserve in larvae as they approached the time of first feeding.     

饲料蛋白质水平对拉萨裸裂尻鱼幼鱼肌肉氨基酸及蛋白质代谢的影响

本实验旨在探索饲料蛋白质水平对拉萨裸裂尻鱼(Schizopygopsis younghusbandi Regan)幼鱼肌肉氨基酸及蛋白质代谢的影响。选用初始体重为(22.42±0.56) g 的拉萨裸裂尻鱼 540 尾,随机分成 6 组,每组 3 个重复,每个重复30 尾鱼,分别投喂蛋白质水平为 20%、25%、30%、35%、40%、45%的实验饲料,养殖时间为 60 d。结果表明:随着饲料蛋白水平的增加,拉萨裸裂尻鱼氮摄入量(NI)和绝对氮摄入量(ANI)均呈逐渐升高的变化趋势。氮沉积(ND)、蛋白质效率(PER)、净蛋白质利用率(NPU)均呈先升高后降低的变化趋势。肌肉总必需氨基酸(TEAA)、总呈味氨基酸(TFAA)、总非必须氨基酸(TNEAA)、总氨基酸(TAA)随饲料蛋白的升高呈先升高后趋于稳定的变化趋势。饲料蛋白水平超过 30%后,血氨(ammonia)、尿素氮(urea)、白蛋白(ALB)显著升高。总蛋白(TP)在饲料蛋白低于 35%时呈逐渐升高的变化趋势。拉萨裸裂尻鱼肝脏谷丙转氨酶 ALT、谷草转氨酶 AST 均呈先升高后趋于稳定的变化趋势,血清 ALT呈先降低后升高的变化趋势,血清 AST在饲料蛋白含量高于 35%后显著升高。综合考虑饲料蛋白利用率、血液总蛋白含量、肝脏及血清转氨酶活性,建议拉萨裸裂尻鱼饲料蛋白含量为 30%~35%。     

Nitrogenous excretion in juvenile turbot, Scophthalmus maximus (L.), under controlled conditions

Laboratory experiments on juvenile turbot. Scophthalmus maximus (L.), were carried out under controlled temperature (12°C) and feeding regimes in a flow-through system. Monitoring of total ammonia nitrogen (TAN) and urea nitrogen (Urea-N) was performed through continuous sampling of the effluent sea water. The effects of ingested nitrogen levels on TAN and Urea-N daily and hourly excretion rates were studied. Strong relationships were found between ingested nitrogen, and both TAN and Urea-N excretion for both daily and hourly maximum excretion rates. Turbot showed a low metabolic activity, confirmed by low excretion levels. Daily patterns for TAN and Urea-N production were different, suggesting specific physiological phenomenon for urea excretion mechanisms. The question of whether turbot are partly ureotelic or ureogenic is underlined.     

Physico‐chemical characterization and in vitro digestibility of commercial feeds used in weaning of marine fish

Six different commercial feeds used in weaning and postweaning of marine fish were analysed from the perspective of some of their main physico‐chemical features and in vitro digestibility. While the range of particle sizes in dry state closely agreed to those claimed by the producing companies, dramatic changes were observed after suspension in water. Sedimentation rates measured in some of the feeds with higher particle sizes may be too high to ensure intake by young fish. The label claims of the products overestimate the amount of total protein but underestimate that of lipids. The analysis of amino acid profiles evidenced important differences in total amino acid composition between feeds of the same company but a great similitude in the profile of free amino acids (FAA). The evaluation of leaching evidenced that the fraction of FAA, which represented <2% of dry matter of the feeds, is quickly released in water, acting mainly as attractants. The in vitro digestibility assay evidenced significant differences in the bioaccessibility of the nitrogen fraction in some of the feeds by the digestive proteases of the sea bream (Sparus aurata).     

Changes in lipid classes,fatty acids,protein and amino acids during egg development and yolk‐sac larvae stage in brill (Scophthalmus rhombus L.)

Brill (Scophthalmus rhombus L.) is a flatfish considered of special interest for aquaculture diversification, but the high mortality observed during the early larval rearing is the main obstacle to commercial culture. The objective of this study was to contribute to the knowledge of nutrient utilization of early‐hatched larvae, characterizing the changes in lipid and protein contents during embryogenesis and the yolk‐sac larval stage of S. rhombus. Total lipid, lipid classes and fatty acid contents remained constant during embryogenesis and yolk‐sac larval development, except for phosphatidylethanolamine and phosphatidylinositol, which increased in quantity during the yolk‐sac larval stage. On the other hand, total protein (including non‐protein nitrogen) and amino acids decreased their contents in both periods, especially at hatching. The decrease only in the serine, glutamic acid, proline and lysine contents during embryogenesis suggests a selective use of amino acids during this phase. Unlike embryogenesis, amino acids loss during hatching appears to be non‐selective, and almost all amino acids (essential and non‐essential) decreased. Our results suggest that there is higher catabolism of protein vs. lipid during embryogenesis and the yolk‐sac larval stage of S. rhombus.