Changes in protein-bound and free amino acids in the muscle of the tiger prawn Penaeus esculentus during starvation

Using the starvation technique, changes in protein and free amino acids were examined in Penaeus esculentus Haswell collected from Moreton Bay, Australia, by trawling in 1985. Prawns of 17.7±0.26 g wet weight were held at 25°C until 2 d after moulting. Groups of seven or eight were then starved fro 5, 10, or 15 d, with appropriate control groups. At the end of each period, ecreted amino acids were collected for 24 h and whole-muscle amino acids and free amino acids (FAA) g^-1 in each prawn were analysed. Concentrations of whole-muscle amino acids showed only minor changes with starvation, but concentrations of many of the FAA changed significantly. Total FAA averaged 1 182±45 mol g^-1 dry weight. Individual FAA, in order of abundance, were glycine, arginine, proline, taurine, threonine, hydroxyproline, alanine, glutamic acid, valine, aspartic acid and lysine; the remaining FAA each contributed <0.2% of the total. Only taurine and alanine did not show significant changes with starvation. Concentrations of glycine, arginine, hydroxyproline, glutamic and aspartic acid increased, while those of proline, threonine, valine and lysine decreased with starvation, that of proline approaching zero after 15 d starvation. Excreted amino acid-nitrogen represented <2% of excreted ammonianitrogen ornithine being the most abundant (35%), followed by leucine (22%) and lysine (17%). The relative abundance of excreted amino acids did not correspond with those of the FAA. It is suggested that, as starvation progresses, the muscle protein is progressively hydrolysed, but with the remaining muscle maintaining its amino acid composition. The liberated amino acids enter the FAA pool and become available for energy production. Proline may have an important role as an energy source, but the ability to synthesise proline may be limited, and thus the artificial food of penaeid prawns may be improved by its addition.     

The essential amino-acid requirements of the prawnPalaemon serratus. The growth of prawns on diets containing proteins of different amino-acid compositions

Two specimens ofPalaemon serratus Pennant (of about 4 g wet weight) were each given intrahaemocoelic injections of [U-¹⁴C] acetate. They were maintained under controlled conditions, and the release of¹⁴CO₂ and other labelled metabolites was followed for 6 days. The prawns were then sacrificed and fractionated into trichloroacetic acid soluble compounds, lipids, nucleic acids and a protein/chitin residue. Little or no¹⁴C had been incorporated into arginine, methionine, valine, threonine, isoleucine, leucine, lysine, histidine, phenylalanine and tryptophan. It was inferred thatP. serratus cannot synthesize these amino acids from ordinarily available materials, and that the prawns have an absolute dietary requirement for them. The glucosamine isolated from the protein/chitin residue contained more radioactivity than any of the other compounds examined. This implies a rapid turnover of glucosamine and chitin inP. serratus. The gain in weight of prawns fed on compound diets containing freeze-dried cod muscle was about 70% that of prawns fed on fresh mussel mantle. Predigestion of freeze-dried cod muscle with proteolytic enzymes did not significantly enhance its nutritional valve as assessed by increases in weight. Prawns fed two other compounded diets containing proteins of low nutritional value (gelatin and zein, respectively) grew at about 1/5 the rate of prawns fed fresh mussel mantle. Supplementation of the gelatin and zein in these diets with tryptophan, and with lysine plus tryptophan, respectively, failed to augment their nutritional value significantly.     

Assimilation of dietary free amino acids,peptides and protein in post-larval Atlantic halibut (<Emphasis Type="Italic">Hippoglossus</Emphasis><Emphasis Type="Italic">hippoglossus</Emphasis>)

With the aim of improving the formulation of diets for early stages of marine fish, post-larval Hippoglossus hippoglossus (56 days after first feeding; 23 mg dry weight) were tube-fed liquid diets of radiolabelled L-(¹⁴C) free amino acids (FAA diet), peptides (PEPT diet) and protein (PROT diet), in order to compare the kinetics of transfer from gut into the liver and carcass. The test diets were prepared using equal concentrations of amino acids (AA) and fed as a single pulse (0.3 µl, 2.2 µg AA). The PROT diet consisted of methylated-¹⁴C-labelled (mainly lysine) bovine serum albumin (BSA), the PEPT diet was the PROT diet digested with trypsin and the FAA diet consisted of an amino acid mixture based on BSA composition with added ¹⁴C-lysine. The common labelling with ¹⁴C-lysine permitted a comparison of the three diets. Based on dissection and distribution of the tracer in each compartment, the FAA diet was almost completely absorbed 1 h after the tube-feeding; PEPT and PROT diets required more time before they were processed by the digestive tract. The FAA diet was absorbed about eight times as fast as protein and about six times as fast as peptides from the gut. A low retention of label in the liver for all diets at all sampling times suggests that the liver does not store the AA at these absorption rates. The FAA diet had a higher assimilation efficiency (about 90%) than the PEPT diet (about 12%) and PROT diet (about 32%). FAAs, thus, appear to be superior to protein and peptides as a dietary source of amino acids in post-larval Atlantic halibut.     

Seagrass and algal beds as nursery habitats for tiger prawns (Penaeus semisulcatus and P. esculentus) in a tropical Australian estuary

We evaluated the importance of seagrass and algae to two species of tiger prawns (Penaeus semisulcatus and P. esculentus) by detailed sampling at four sites (two seagrass, two algae) in the Embley River estuary, and through sampling 26 sites in 7 adjacent estuaries at one time of year. Samples of tiger prawns were collected in the Embley River estuary with a small beam trawl at night every 2 wk from September to May for 2 yr (1990 to 1992). The two seagrass sites, which were 11 and 13 km from the river mouth, showed less seasonal variation in salinity than the two algal sites, which were 15 and 20 km from the river mouth. The algal beds at the two upstream sites almost disappeared during the wet season, but the biomass of seagrass did not change significantly between the wet and dry seasons. The grooved tiger prawn (P. semisulcatus), the main species at all sites, comprised 88% of the total tiger prawn catch over the two years. They were found at all sites during the pre-wet season, but after the onset of the wet season, they disappeared along with the algae, from the upstream sites. The brown tiger prawn (P. esculentus) was found almost exclusively (97% of the total catch) on the seagrass sites downstream. In the study of several estuaries, juvenile P. semisulcatus were caught at all 26 sites, and P. esculentus were caught in much smaller numbers, at 16 sites. Approximately equal numbers of P. semisulcatus were caught in seagrass and algal beds in the pre-wet season. Very few individuals >10 mm carapace length of either species, were caught. The results from this study highlight the importance of algal beds during the pre-wet season as nursery areas for one species of tiger prawn (P. semisulcatus).     

Seasonal changes in nucleic acids,amino acids and protein content in juvenile Norway lobster (<Emphasis Type="Italic">Nephrops norvegicus</Emphasis>)

The objective of this study was to describe the seasonal variations in nucleic acid contents and amino acid profiles in the muscle of juvenile Nephrops norvegicus. RNA and protein contents, and RNA:protein and RNA:DNA ratios varied significantly between seasons, being highest in spring and lowest in autumn/winter ( P<0.05). Though DNA content increased significantly from autumn to summer ( P<0.05), protein:DNA ratio did not show significant seasonal variations ( P=0.05). In respect to protein-bound amino acid content (BAA), a significant increase was observed from winter to summer ( P<0.05). Both essential (EAA) and non-essential amino acids (NEAA) increased significantly (27.6% and 27.8%, respectively; P<0.05), mainly due to the considerable increase in arginine and proline (59.1% and 225.2%, respectively; P<0.05). A significant decrease was observed in the free amino acid content (FAA) from winter to summer ( P<0.05); and a higher percentage decrease occurred in free non-essential (FNEAA; 27.9%) in comparison to free essential amino acids (FEAA; 21.8%). The significant increase in RNA and BAA contents from winter to spring may be related to protein synthesis. On the other hand, the lowest values obtained in winter may be due to a reduction in feeding activity; in this period the muscle protein must be progressively hydrolysed, which is evident with the higher FAA content. The liberated amino acids enter the FAA pool and become available for energy production.     

Osmoregulation of the grass shrimp Palaemonetes pugio Exposed to polychlorinated biphenyls (PCBs).

Glycine was the most abundant free amino acid (FAA) in abdominal muscle of grass shrimp Palaemonetes pugio, accounting for over 50% of the total pool. Arginine, alanine, proline, taurine and serine were also present at relatively high concentrations. Following transfer from 17%. to 2 and 32%. S, new steady-state levels of total FAA were observed at 72 h. Total FAA and the sum of glycine, alanine and proline exhibited a positive correlation with salinity. Exposure to Aroclor 1254 did not have appreciable effects on total FAA levels, indicating that disruption of intracellular osmoregulation was not a major consequence of PCB toxicity. However, changes in individual amino acid concentrations in exposed shrimp were reflective of an altered metabolic state. Glycine, which did not show changes immediately following exposure, underwent large decreases after transfer to PCB-free water and indicated a delayed effect of PCB exposure. A correlation between salinity and the sum of glycine, alanine and proline did not exist in exposed shrimp.     

Effects of nutritional history on nitrogen assimilation in congeneric temperate and tropical scleractinian corals

The nutritional history of corals is known to affect metabolic processes such as inorganic nutrient uptake and photosynthesis, but little is known about how it affects assimilation efficiency of ingested prey items or the partitioning of prey nitrogen between the host and symbiont. The temperate scleractinian coral Oculina arbuscula and its tropical congener Oculina diffusa were acclimated to three nutritional regimes (fed twice weekly, starved, starved with an inorganic nutrient supplement), then fed Artemia nauplii labeled with the stable isotope tracer ¹⁵N. Fed corals of both species had the lowest assimilation efficiencies (36–51% for O. arbuscula, 38–57% for O. diffusa), but were not statistically different from the other nutritional regimes. Fed and starved corals also had similar NH₄⁺ excretion rates. This is inconsistent with decreased nitrogen excretion and reduced amino acid catabolism predicted by both the nitrogen recycling and conservation paradigms. In coral host tissue, ~90% of the ingested ¹⁵N was in the TCA-insoluble (protein and nucleic acids) and ethanol-soluble (amino acids/low molecular weight compounds) within 4 h of feeding. The TCA-insoluble pool was also the dominant repository of the label in zooxanthellae of both species (40–53% in O. arbuscula, 50–60% in O. diffusa). However, nutritional history had no effect on the distribution of prey ¹⁵N within the biochemical pools of the host or the zooxanthellae for either species. This result is consistent with the nitrogen conservation hypothesis, as preferential carbon metabolism would minimize the effects of starvation on nitrogen-containing biochemical pools.Communicated by P.W. Sammarco, Chauvin     

Free amino acid and protein contents of start-feeding larvae of turbot (Scophthalmus maximus) at three temperatures

The contents of free amino acids (FAA) and total protein, together with growth and gut-content, of turbot (Scophthalmus maximus L.) larvae reared at 14, 18 and 22 °C were studied from first-feeding to approximately 140 effective day-degrees post hatch (D_eff ^∘). Artemia franciscana nauplii and two species of rotifers were used as prey. Protein content accounted for about 42 and 26% of dry body mass in the A. franciscana nauplii and the rotifers, respectively. The FAA pool constituted 5.6 and 4.8% of the total amino acids in the same animals. The dry body mass of turbot larvae was exponentially related to D_eff ^∘. Protein and FAA contents were linearly related to dry body mass, and were independent of rearing temperature between 14 and 18 °C. At the end of the experiment, however, turbot larvae at 22 °C had lower gut content values, retarded growth rates, and decreased FAA contents and concentrations. Thus, at this high temperature, turbot larvae seem unable to catch and ingest sufficient prey, or to sustain an amino acid assimilation rate from the intestine sufficient to meet metabolic demands. Received: 2 January 1997 / Accepted: 25 September 1998     

The mechanism of crustacean salinity tolerance: Cell volume regulation by K+ and glycine effluxes

The regulation of muscle fiber K⁺ and free amino acid (FAA) concentrations during hypoosmotic stress was investigated in the moderately euryhaline crab Cancer irroratus. After 6 h of exposure to 60% ASW, muscle fiber K⁺ concentration declined from 185 mM to 140 mM. Following this, the blood glycine levels began to increase, indicating an FAA efflux from the cells. These data indicate that both muscle fiber K⁺ and FAA contribute to cell volume regulation in C. irroratus. The early release of K⁺ limits the initial rate of cell hydration. The subsequent efflux of glycine accounts for the volume regulation response of the muscle fibers. The cell volume regulatory system of C. irroratus is a coordinated use of both inorganic ions and FAA.     

Free amino acids and energy metabolism in eggs and larvae of seabass, Lates calcarifer

Contents of free amino acids (FAA), protein and ammonium ions together with rates of ammonia excretion and oxygen consumption were measured in order to study the role of FAA as an energy substrate in developing eggs and larvae of seabass (Lates calcarifer) maintained in seawater (30 ppt) at 28 °C without feeding. Initially eggs contained 25.3 nmol ind⁻¹ of FAA of which 21.5 nmol was rapidly utilised by the developing eggs and larvae during the period up to 40 h post spawning (PS) when nearly all the yolk had been resorbed. During the same period, a net increase in protein content of 1.7 μg ind⁻¹ was observed, indicating that the major part of the amino acids lost from the free pool had been polymerised into body proteins. Assuming that the balance of the FAA after protein synthesis was used entirely for energy metabolism, FAA appeared to be an important energy substrate during the embryonic stages (2 to 16 h PS); after hatching, the contribution of FAA to energy metabolism was less significant. From 50 h PS until the end of the study period at 100 h PS, amino acids derived from somatic protein were used for energy metabolism. For the overall period from just after spawning up to 100 h PS, the data indicate that ca. 14% of the total aerobic energy metabolism was derived from amino acid catabolism. Received: 26 September 1997 / Accepted: 1 April 1998     

Larval durations and recruitment patterns of two Caribbean gobies (Gobiidae): contrasting early life histories in demersal spawners

The population dynamics of small tiger prawns (Penaeus esculentus and P. semisulcatus) were studied at three sites around north-western Groote Eylandt, Gulf of Carpentaria, Australia, between August 1983 and August 1984. Seagrasses typical of open-coastline, reef-flat and river-mouth communities were found in the shallow depths (2.5 m) at these sites. The temperature and salinity of the bottom waters did not differ among the shallowest depths of the three sites and mean values at night ranged from 21.9 to 32.0 °C, and from 30.1 to 37.5% S. Data from fortnightly sampling with beam trawls showed that virtually all post-larvae (90%) were caught in the intertidal and shallow subtidal waters (2.0 m deep). At one site, where the relationship between seagrass biomass, catches and depth could be studied in detail, high catches were confined to seagrass in shallow water, within 200 m of the high-water mark. This was despite the fact that seagrass beds of high biomass (>100 g m^-2 between August and February) were found nearby, in only slightly deeper water (2.5 m). It is likely, therefore, that only the seagrass beds in shallow waters of the Gulf of Carpentaria act as important settlement and nursery areas for tiger prawns. In general, catches of tiger prawn postlarvae (both P. esculentus and P. semisulcatus) and juvenile P. esculentus on the seagrass in the shallowest waters at each site were higher in the tropical prewet (October–December) and wet (January–March) seasons than at other times of the year. Juvenile P. semisulcatus catches were highest in the pre-wet season. While seasonal differences accounted for the highest proportion of variation in catches of tiger prawn postlarvae and juvenile P. semisulcatus, site was the most important factor for juvenile P. esculentus. In each season, catches of juvenile P. esculentus were highest in the shallow, open-coastline seagrass, where the biomass of seagrass was highest. The fact that the type of seagrass community appears to be more important to juvenile P. esculentus than to postlarvae, suggests that characteristics of the seagrass community may affect the survival or emigration of postlarval tiger prawns. Few prawns (<10%) from the seagrass communities in shallow waters exceeded 10.5 mm in carapace length. Despite the intensive sampling, growth was difficult to estimate because postlarvae recruited to the seagrass beds over a long period, and the residence times of juveniles in the sampling area were relatively short (8 wk).     

Nitrogen regeneration by the surf zone penaeid prawn Macropetasma africanus

Nitrogen excretion of individual Macropetasma africanus (Balss) from an exposed beach/surf zone in Algoa Bay, South Africa was monitored under laboratory and field conditions in relation to body mass, temperature and feeding during 1985. Excretion rate experiments were performed on starved prawns at 15°, 18°, 20° and 25°C, as well as on individuals fed on four different diets (mussel, fish, shrimp and natural diet) at 15° and 20°C. The ratios of the excreted compounds to total nitrogen excreted were similar for the four diets despite differences in their nitrogen content and in the amount of food consumed. At 15° and 20°C, ammonia excretion rates of fed individuals were four to seven times higher than in starved prawns. the excretion rates were not correlated with nitrogen content of diets. M. africanus recycles 1 557 g NH₄–N per metre strip per year or 1 832 g total nitrogen m^-1 yr^-1, which constitute 12 and 14%, respectively, of total phytoplankton requirements of the surf zone. This study indicates that large motile crustaceans, when abundant, can play an important role in nutrient recycling in turbulent marine environments.     

Estimating the influence of prawn stocking density and seagrass type on the growth of juvenile tiger prawns (Penaeus semisulcatus): results from field experiments in small enclosures

Fine mesh enclosures (0.9 m² in basal area, 1 m high, with 100 µm mesh) and a jet-net retrieval system were developed to test the influence of juvenile prawn stocking density on growth rates in (1) different months (April and October/November) and (2) different types of intertidal seagrass beds in the Embley River estuary of tropical Australia. Small juvenile tiger prawns (3-6 mm in carapace length, CL) were stocked in enclosures at densities of 4-32 prawns per enclosure (4.4-35.5 prawns m^-2) on a high biomass seagrass bed (about 70 g m^-2 of mostly Enhalus acoroides) and one with low biomass (about 10 g m^-2 of mostly Halodule uninervis). After 2-3 weeks in the enclosures, recovery rates, and hence possibly survival, were greater on the high biomass Enhalus than on the low biomass Halodule. However, not all fish and crustaceans could be excluded from the enclosures. Growth rates were twice as fast on the high biomass Enhalus than on the low biomass Halodule. It is likely that the high biomass Enhalus, with its greater surface area, supported more epiphytic flora and fauna and reduced the potential for interference competition between prawns, compared with the low biomass Halodule. Growth rates on Enhalus were significantly faster at a stocking density of 4 prawns per enclosure (1.3 mm CL week^-1) than at a stocking density of 16 and 32 prawns per enclosure (both 0.8 mm CL week^-1), and did not differ significantly between April and October/November (temperatures were about 30°C at both times). The mean growth rate at 8 prawns per enclosure (1.1 mm CL week^-1) did not differ significantly from those at 4, 16 and 32 prawns per enclosure. These results from two seagrass beds suggest that the carrying capacity for juvenile tiger prawns was greater in the high biomass Enhalus than the low biomass Halodule bed.     

Nitrogen and energy loss via nonfaecal and faecal excretion in the marine teleost Lithognathus lithognathus

Nitrogen excretion and assimilation efficiencies of individual Lithognathus lithognathus (Cuvier 1830), a marine teleost from high energy surf zones in Algoa Bay, South Africa, were determined under laboratory conditions in 1985. Nonfaecal-nitrogen excreted by starved and fed L. lithognathus consists mainly of ammonia with urea and amino acids as secondary excretory products. Ammonia excretion rates were temperature dependant with the excretion rate of starved fish significantly lower than those of fed fish, at all three experimental temperatures. The mass component b of the mass/ammonia excretion equation was temperature independent and ranged from 0.651 to 0.700 and 0.589 to 0.635 for starved and fed fish respectively. The mean percentage of food energy lost via dissolved nonfaecal excretory products (exogenous plus endogenous) was 6.11±6.07%. Assimilation efficiencies ranged from 70.75 to 99.29% for dry matter and from 95.72 to 99.58% on an energy basis. The combined nonfaecal and faecal energy loss was calculated at 11.87% of the ingested energy. The benthic feeding ichthyofauna recycle 255 g total nitrogen per metre strip per year which constitutes 2% of total phytoplankton nitrogen requirements of the surf zone.     

Stimulatory effect of ingested protein and/or free amino acids on the secretion of the gastro-endocrine hormone cholecystokinin and on tryptic activity, in early-feeding herring larvae, Clupea harengus

In the larvae of many marine teleosts, the stomach is absent until they approach or attain metamorphosis. Consequently, the formation of chyme containing specific free amino acids from the gastric digestion of protein, which are believed to be signals initiating the release of the digestive hormone cholecystokinin (CCK), is lacking. CCK, when secreted into the blood circulation from specialized intestinal cells, stimulates gallbladder motility and is a key factor causing the release of pancreatic digestive enzymes into the gut lumen. Using first-feeding Atlantic herring larvae (Clupea harengus) as a model, the aim of the present study was to determine if a CCK response together with tryptic activity could be elicited in larvae ingesting dietary protein and/or FAA. Larvae were tube fed single lamellar liposome vesicles (SLV) containing: (1) physiological saline (PS), (2) bovine serum albumin (BSA), (3) specific free amino acids (FAA), or (4) a ratio (1:1) of BSA and FAA. The CCK and trypsin levels were then assayed (radio-immunoassay) at 0, 15, 60 and 120 min after tube feeding. A marked CCK response was elicited in all treatments compared to the PS control at 15 and 30 min and was significant (p<0.05) at 120 min after tube feeding. Larvae tube fed the FAA treatment exhibited CCK levels that increased linearly from 1.6 to 5.6 fmol mg^-1 dry weight (DW) after 2 h of digestion, although this response was below the BSA and BSA:FAA treatments. The BSA and BSA:FAA treatments, after 15 min of digestion, showed a rapid CCK increase, over the PS and the FAA liposome treatments, to 8.1 and 5.4 fmol mg^-1 DW, respectively. At the end of the assay, BSA and BSA:FAA demonstrated similar levels (10.2 and 9.2 fmol mg^-1 DW, respectively). Larvae tube fed the PS control or the FAA liposome treatment did not demonstrate any appreciable increase in tryptic activity during the 2 h digestion period (0.03-0.071 and 0.03-0.048 mU mg^-1 DW, respectively). In contrast, the BSA:FAA treatment increased from 0.03-0.148 mU mg^-1 DW 1 h after feeding, which was significantly (p<0.05) higher than the PS and FAA liposomes, and then decreased markedly (0.085 mU mg^-1 DW) after 2 h of digestion. The larvae tube fed BSA liposomes, however, demonstrated steadily increasing tryptic activity throughout the sampling period, attaining 0.255 mU mg^-1 DW after 2 h, which was significantly (p<0.05) more than all the other treatments. The results showed that ingested liposomes containing FAA or the protein BSA or a combination of these two nutrients effectively stimulated CCK production in first-feeding herring larvae. In contrast, liposomes containing only physiological saline did not elicit a CCK response. In addition, liposomes containing BSA stimulated tryptic activity in herring larvae, which was not observed in fish fed liposomes that included only FAA or PS. This suggests that a suitable protein substrate is required to regulate protein digestion.     

Relative importance of benthic microalgae,phytoplankton, and mangroves as sources of nutrition for penaeid prawns and other coastal invertebrates from Malaysia

Carbon, sulfur, and nitrogen stable isotope ratio techniques were used in 1989 and 1990 to evaluate the relative importance of algae and of mangrove detritus in the nutrition of two penaeid prawn species on the west coast of Peninsular Malaysia. Mangrove detritus was found to contribute to the nutrition of juvenile Penaeus merguiensis de Man living within tidal creeks, but not to adult P. merguiensis and juvenile and adult Parapenaeopsis sculptilis (Heller) captured offshore. Results from radiotracer feeding studies, with refractory ¹⁴C mangrove lignocellulose as the food source, indicated that juvenile P. merguiensis from tidal creeks assimilated mangrove carbon with an efficiency of 13.4%. This did not differ significantly (P=0.05) from the assimilation efficiencies of juvenile and adult P. sculptilis living offshore (10.0 and 10.9%, respectively); these values were significantly higher (P<0.001) than for adult P. merguiensis (2.1%). Thus, the differential contribution of mangrove material to the nutrition of prawns in the tidal creeks and offshore was not related to differences in the prawn's ability to utilize detritus. Instead, our analysis of C stable isotopes in sediments indicated that mangrove detritus was generally more abundant within the tidal creeks than offshore. Juvenile prawns in the tidal creeks may also utilize mangrove material indirectly by feeding on small detritivorous invertebrates. Stable isotope analyses suggest that benthic microalgae constitute the other major dietary component for prawns living in tidal creeks. Prawns offshore were utilizing mainly phytoplankton-based material.     

Amino acid metabolism and protein turnover in larval turbot (Scophthalmus maximus) fed natural zooplankton or Artemia

The present paper studied the influence of different food regimes on the free amino acid (FAA) pool, the rate of protein turnover, the flux of amino acids, and their relation to growth of larval turbot (Scophthalmus maximus L.) from first feeding until metamorphosis. The amino acid profile of protein was stable during the larval period although some small, but significant, differences were found. Turbot larvae had proteins which were rich in leucine and aspartate, and poor in glutamate, suggesting a high leucine requirement. The profile of the FAA pool was highly variable and quite different from the amino acid profile in protein. The proportion of essential FAA decreased with development. High contents of free tyrosine and phenylalanine were found on Day 3, while free taurine was present at high levels throughout the experimental period. Larval growth rates were positively correlated with taurine levels, suggesting a dietary dependency for taurine and/or sulphur amino acids. Reduced growth rates in Artemia-fed larvae were associated with lower levels of free methionine, indicating that this diet is deficient in methionine for turbot larvae. Leucine might also be limiting turbot growth as the different diet organisms had lower levels of this amino acid in the free pool than was found in the larval protein. A previously presented model was used to describe the flux of amino acids in growing turbot larvae. The FAA pool was found to be small and variable. It was estimated that the daily dietary amino acid intake might be up to ten times the larval FAA pool. In addition, protein synthesis and protein degradation might daily remove and return, respectively, the equivalent of up to 20 and 10 times the size of the FAA pool. In an early phase (Day 11) high growth rates were associated with a relatively low protein turnover, while at a later stage (Day 17), a much higher turnover was observed. Received: 19 March 1997 / Accepted: 14 April 1997     

Major osmolyte changes during oocyte hydration of a clupeocephalan marine benthophil: Atlantic herring (<Emphasis Type="Italic">Clupea harengus</Emphasis>)

The major inorganic and organic osmolytes responsible for hydrating the oocytes during pre-ovulatory meiotic maturation in autumn- and spring-spawning stocks of Atlantic herring are examined. Despite the ovulated eggs of spring-spawning herring being 1.6- to 2-fold larger than the autumn-spawning stock, the GSI (27 ± 3%) and degree of oocyte hydration (70–72% water) were similar. Normalising the data with respect to dry mass revealed that the physiological mechanisms underlying the maturational influx of water were the same for both classes of egg. Cl⁻, K⁺ and P_i together with a small pool of free amino acids (FAA) represented the driving forces for oocyte hydration. K⁺ (autumn and spring) and P_i (spring) maintained their concentrations in the ovulated eggs, while all other ions, including Cl⁻, Na⁺, NH₄ ⁺ and Mg²⁺ were significantly diluted. In contrast the FAA concentration increased during the hydration process. Amongst the inorganic ions, Cl⁻ showed the greatest increase in the ovulated eggs. The FAA content doubled from 1.5 to 3.3% of dry mass during oocyte hydration and accounted for 29% of the calculated ovoplasmic osmolality in the ovulated eggs from both autumn- and spring-spawners. This significant osmotic effect of the small pool of FAA was due to the low water content of the benthic eggs. The differential movement of the inorganic and organic osmolytes that underly oocyte hydration in Atlantic herring are discussed in relation to current models of transmembrane ion flux.     

Changes in metabolic substrates during early development in anchoveta Engraulis ringens (Jenyns 1842) in the Humboldt Current

We assessed the ontogenetic changes in protein content and free amino acids (FAA) in eggs and early larvae of Engraulis ringens (anchoveta) off central Chile on different dates during the spawning season. On all sampling dates, a reduction in embryonic yolk-sac volume, proteins and FAA concentrations occurred during development. Protein electrophoresis (SDS–PAGE) of eggs and larvae showed at least 22 protein bands: 11 were consumed early and not detected after hatching. The proportion of essential FAA (EFAA) was higher than the proportion of non-essential FAA (NEFAA) in early eggs and in 7 day-old larvae (82.5-73% EFAA respectively). During egg development, the FAA pool was dominated by leucine, alanine and lysine, three amino acids contributing 35–44% of the total FAA in eggs. During larval development, histidine was the most abundant FAA. In July, total FAA constituted 13–18% of the egg dry weight. A similar proportion (45–51%) occurred in July between protein plus FAA and total lipids. The differences in egg size during the spawning season along with variability in batch composition suggests that the female spawning condition is a major factor determining egg quality and early offspring success.     

Influence of cadmium accumulation and dietary status on fatty acid composition in two colour forms of shore crabs, Carcinus maenas

 The effects of cadmium exposure and dietary status on cadmium accumulation, fatty acid (FA) content and profiles were investigated in two colour forms of the shore crab Carcinus maenas. Groups of shore crabs were either starved or fed with blue mussels, Mytilus edulis, during a 40 d exposure period to 2 or 6 μM Cd²⁺ (as CdCl₂). Starved green individuals accumulated more cadmium in haemolymph and hepatopancreas than did red crabs and green crabs fed during the experiments. In the red colour form, no difference in cadmium accumulation was observed between starved and fed individuals. In both colour forms, hepatopancreas contained more FA than gills and muscle. The FAs often present in the largest amounts in the tissues were 16:0, 16:1ω7, 18:1ω7, 18:1ω9, 20:4ω6, 20:5ω3 and 22:6ω3. However, saturated (SAFA) and mono-unsaturated fatty acids (MUFAs) were dominant in hepatopancreas, whereas poly-unsaturated fatty acids (PUFAs) were dominant in gills and muscles. At the beginning of the experiment, the total FA content in the hepatopancreas was 111.6 mg g⁻¹ (dry weight) for red crabs and 78.4 mg g⁻¹ for green shore crabs. During the experiment, however, the FA content decreased in red crabs. This decrease was more pronounced for starved individuals than for fed individuals. Also, the decrease in FA content was more pronounced in crabs exposed to 6 μM cadmium compared to crabs exposed to 2 μM or crabs not exposed to cadmium. No change in FA content was observed in green shore crabs, irrespective of diet and cadmium exposure. For both colour forms, no change in FA content was observed for gills and muscle. In red crabs, a decrease was observed for all FAs in the hepatopancreas. This decrease, however, was more pronounced for SAFAs and MUFAs than for PUFAs, indicating that the metabolism of FAs during starvation and cadmium exposure is selective. The experiments indicate that green colour forms of shore crabs are more tolerant of natural stress such as starvation and anthropogenic stress, e.g. cadmium exposure, than are red colour forms of shore crabs. Received: 23 September 1999 / Accepted: 29 April 2000