Studies on the nutrition of marine flatfish. Growth of the plaice Pleuronectes platessa on diets containing proteins derived from plants and other sources

The growth of 0-group Pleuronectes platessa L. on diets containing plant proteins was examined. Diets were made acceptable to the fish by including in them cod flesh which had been predigested with proteolytic enzymes, as well as a proportion of air-dried cod meal. A leaf-protein concentrate was fed at 6 levels ranging from 0 to 63 % of the total dietary protein; B.P. protein concentrate (Toprina) and soyabean meal were examined at a single level — about 45 % of the dietary protein. The remainder of the protein in these diets was animal protein, cod meal or cod flesh. Analysis of the weight gains after a 12 week feeding period and of the growth curves showed that, as the proportion of leaf protein in the diet increased, the growth rate diminished, the effect becoming more pronounced when leaf protein reached 40 % or more of the dietary protein. Growth on the diet containing B.P. protein concentrate was superior to that on the diet containing soyabean meal and was comparable with diets containing low levels of leaf protein. The protein efficiency ratios (PER= g live weight gain/g protein fed) for the different diets showed similar trends to the growth rates. Diets containing 40 % or less of leaf protein and the B.P. protein diet had PERs of 1.4 to 1.5. The soyabean meal diet and diets containing more than 40 % of the protein as leaf protein had PERs of 1.0 or less. The results are considered in the context of production diets for flatfish.     

Effects of various diets on the nutritional value of Artemia sp. as food for the prawn Penaeus monodon

The nutritional value of Artemia sp. as food for marine fish and crustacean larvae has been linked to the level of its polyunsaturated fatty acid (PUFA) content. Experiments in August 1984 were conducted to determine the effects of various artificial diets and algae on fatty acid composition of PUFA-deficient Artemia sp. (Utah GSL strain) and their resulting value as food for postlarvae of the prawn Penaeus monodon (Fabricius). Nauplii of the brine shrimp were grown on extracts of corn, copra, soybean and rice bran containing precursors (C18) to long-chain PUFA and also on algal species containing different levels of long-chain PUFA (C20). The nauplii were then used as food for P. monodon postlarvae. The results revealed that absence of C20 polyunsaturates from the feeds and their presence in the algae were reflected in the polyunsaturated fatty acid content of the tissues of Artemia sp. When fed with brine shrimp fed on algae, P. monodon displayed better postlarval survival and significantly higher growth; related to the content of polyunsaturated fatty acids in Artemia sp. A practical feeding approach in prawn hatcheries would be to grow Artemia sp. on a cheap diet such as rice bran, and then to enhance its nutritional value with a diet high in PUFA prior to harvesting, in order to improve hatchery production.     

Ammonia excretion by the benthic estuarine shrimp Crangon franciscorum (Crustacea: Crangonidae) in relation to diet

Ammonia excretion of individual Crangon franciscorum Stimpson was monitored in response to ingestion of single meals. The three experimental diets were tubificids, mysids and fish. Ammonia excretion was also monitored for individual shrimp which had been starved. The rate of ammonia excretion was higher for fed than for starved individuals in all cases. Ammonia excretion rates were higher for shrimp which were fed tubificids than those fed the other diets. The rate of excretion was influenced by both weight of the individual and the amount ingested of each diet. Ammonia excretion was influenced by dietary factors other than nitrogen content of the diet or the quantity ingested. The data suggest that field estimates of ammonia excretion based on the excretion rates of starved animals may be underestimates. The recent feeding history of an organism influences the rate of ammonia excretion as well as the relationship between the rate of excretion and weight.     

Responses of hemolymph osmolality and tissue water of Penaeus chinensis Osbeck juveniles subjected to sudden change in salinity

Hemolymph osmolality and tissue water of laboratory-reared Penaeus chinensis Osbeck juveniles (0.83 to 1.86 g) were investigated, after they had been transferred individually from 10, 20, 30 and 40 ppt to 10, 20, 30 and 40 ppt for 0.25, 0.5, 1, 2, 5 and 10 d, respectively. Hemolymph osmolality and tissue water of shrimp were stablilized within 5 d after they had been subjected to a sudden change in salinity from each salinity level. Hemolymph osmolality had a positively linear relationship with medium osmolality. Tissue water decreased with increased medium osmolality, and decreased with increased hemolymph osmolality. The mean (SD) isosmotic point was 703 (8) mOsm kg^–1 which is equivalent to 24.2 (1.0) ppt. P. chinensis juveniles exhibited hyperosmotic regulation in salinities below isosmotic value, and hypoosmotic regulation in those above. The shrimp originally adapted to high salinity levels (30 and 40 ppt) showed less fluctuation of tissue water than those adapted to low salinity levels (10 and 20 ppt) when they were subjected to a sudden change in salinity.     

The iridoid glycoside,catalpol, as a deterrent to the predatorCamponotus floridanus (Formicidae)

Summary We investigated the role of the iridoid glycoside, catalpol, as a deterrent to the predator,Camponotus floridanus. Four laboratory colonies of this ant were offered buckeye caterpillars (Junonia coenia: Nymphalidae) raised on diets with and without catalpol. The same colonies were offered sugar-water solutions containing varying concentrations of catalpol, in both no-choice and choice tests. Regardless of diet, buckeye caterpillars appeared to be morphologically protected from predation by the ants, possibly because of their large spines or tough cuticle. However, buckeyes raised on diets with catalpol had high concentrations of catalpol in their hemolymph; extracts of this high-catalpol hemolymph proved to be an effective deterrent to the ants. When starved ants were not given the choice of food items, they were more likely to consume sucrose solutions that contained 5 mg catalpol/ml or 10 mg catalpol/ml than they were to consume solutions with 20 mg catalpol/ml. When they were given a choice of sugar solution or a sugar solution containing catalpol, the ants avoided solutions with catalpol at any of these concentrations. Ant colony responses to catalpol in sucrose solutions varied considerably over time and among colonies.     

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.     

Diets of calanoid copepods on the West Florida continental shelf: Relationships between food concentration,food composition and feeding activity

Bottle incubations were conducted in March, July/August and October 1992. to measure the daily rations (R) and objectively characterize the diets of the calanoid copepodsEucalanus elongatus, Undinula vulgaris, Centropages velificatus andTemora stylifera from the west Florida continental shelf. Daily rations,R, were clustered around two, order-of-magnitude different means, 1.3 and 11.2% of body C d^–1, representative of quiescent and active feeding modes, respectively. The food concentration at which the transition from quiescent to active mode occurred was influenced by food particle size. In the quiescent mode, diets were dominated by nanoplankton, whereas no food type dominated the diet in the active mode. Selective feeding, defined as a statistically significant difference between the frequency distributions of foods in the diet and environment, occurred in both quiescent and active copepods. However, what appeared to be selective feeding in quiescent copepods could be explained by processes that passively modified the distribution of the diet relative to that of the food supply. Conversely, selective feeding in active copepods apparently resulted from foraging for particles >5 m in diameter in food environments dominated by nanoplankton (<5 m).     

The biological degradation of arginine and glutamic acid in seawater in relation to the growth of phytoplankton

The responses of natural summer coastal plankton communities to low-level additions (10^-5 to 10^-7 M) of arginine and glutamic acid has been followed by in vivo measurement of chlorophyll fluorescence. This technique is capable of detecting a response to a 10^-6 M enrichment under most conditions. The time sequence of the response varied with the amino acid used and with the enantiomeric form of the amino acid. Ammonia and carbon dioxide were liberated before the increase in chlorophyll fluorescence occurred. Liberation of ammonia in a dark bottle from L-arginine was from 75 to 85% of the theoretical yield. Microautoradiography using ¹⁴C L-arginine or L-glutamic acid at 10^-7 M showed heavy labeling associated with fecal pellets and detrital aggregates. Phytoplankton cells were not appreciably labeled. The evidence suggests that bacteria are important in the cycling of these compunds.     

Trophic ecology of the stomiid (Pisces: Stomiidae) fish assemblage of the eastern Gulf of Mexico: Strategies,selectivity and impact of a top mesopelagic predator group

The trophic ecology of the stomiid assemblage (Pisces, Stomiiformes, Stomiidae) in the eastern Gulf of Mexico, a region with physical and biological characteristics typical of oligotrophic low-latitude regimes, was investigated. Over 1400 specimens representing 69 species and 17 genera were examined. Four patterns of feeding were evident among the abundant stomiids: (1) myctophid predation; (2) zooplankton/small micronekton predation; (3) penaeidean shrimp predation; and (4) copepod/micronekton predation. One rare species preyed on cephalopods. Il was concluded that stomiids exhibited a high level of prey-selectivity, particularly considering the broad range of prey types available in the eastern Gulf of Mexico. The absence of numerically dominant potential prey (e.g.Cyclothone spp., sternoptychids) in the diets of piscivorous stomiids is possibly a function of feeding periodicity coupled with stomiid vertical migration. Stomiids may feed at night in the upper 200 m on vertically migrating myctophids while disregarding co-occurring nonmigrating prey during the daytime. Integration of stomiid abundance and diet data suggests that: (1) stomiids are the dominant upper trophic-level predators of the Gulf of Mexico mesopelagial, (2) stomiids inflict the highest predation impact on myctophids in low-latitude midwater ecosystems, and (3) the historic use of predation-avoidance arguments to explain certain mesopelagic phenomena (e.g. vertical migration, ventral photophores) appears to be substantiated by estimates of stomiid predation-impact. The stomiids may serve as key trophic mediators in the transfer of energy from the mesopelagial to the bathyand benthopelagial.     

Isolation and characterization of two actins of the Pacific white shrimp, <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

Two actin genes named actinT1 and actinT2 were isolated and sequenced from the Pacific white shrimp, Litopenaeus vannamei, by screening from a shrimp eyestalk cDNA library. L. vannamei actinT1 cDNA has a 1,128-bp open reading frame encoding for 376 amino acids while L. vannamei actinT2 cDNA has a 1,131-bp open reading frame coding for 377 amino acids. Alignment of the actinT1 and actinT2 cDNA sequences showed that these two actin genes share a sequence identity of 86% at amino acid residues. When compared with actins of several other invertebrate and vertebrate species, the nucleotide sequence of actinT1 is highly homologous (97–100%) with beta-actins, while actinT2 shares 86–95% identity with alpha-actins on the nucleotide level. Phylogenetic analysis and BLAST searches indicated that the ActinT1 protein is identical to crustacean beta-actins, while the ActinT2 protein is highly homologous to crustacean alpha-actins. Constitutive expression of the actinT1 and actinT2 genes were detected by RT-PCR in all adult shrimp organs, including brain, eye-stalk, gill, heart, hemolymph, hepatopancreas, muscle, swimming legs, and stomach, as well as in the shrimp zygote, nauplius, and mysis life stages. These data will facilitate attempts to clone and identify more shrimp genes and constitutive shrimp promoters.     

Respiratory regulation and the role of the blood in the burrowing shrimp Callianassa californiensis (Decapoda: Thalassinidae)

Questions relating to oxygen extraction and hemolymph oxygen transport have been investigated in the burrowing shrimp Callianassa californiensis Dana. The relationship of oxygen consumption to oxygen tension was determined in intermolt and postmolt shrimp. Soft postmolt individuals had higher rates of oxygen consumption and a somewhat lower critical oxygen tension than intermolt specimens. Both oxygen-carrying capacity and hemocyanin content are somewhat lower in postmolt compared to intermolt shrimp. The measured gradient between postbranchial (pericardial sinus) and prebranchial (abdominal sinus) hemolymph oxygen content is small and highly variable, and is not affected by exposure to anoxic stress. Hemolymph pH and buffering capacity are high. The oxygen consumption-oxygen tension relationships and respiratory properties of the hemolymph are discussed in relation to the hypoxic environment associated with the habitat of these crustaceans.Supported in part by Institutional Sea Grant 04-3-158-4 and Public Health Service Grant HL 12326.Communicated by J.S. Pearse, Santa Cruz     

Identification and quantitative analysis of the amino acids present in protein of the brown shrimp Penaeus aztecus

In order to better formulate an artificial shrimp diet, the protein composition of shrimp was assessed and the essential amino acids determined. Penaeus aztecus were treated to remove lipids, carbohydrates and other interfering substances, and the protein was hydrolyzed to its constituent amino acids. The amino acids were then quantitatively analyzed by gasliquid chromatography as their trimethylsilyl derivatives. The essential amino acids were then determined. Shrimp were injected with ¹⁴C-labeled glucose and analyses performed to determine which amino acids then exhibited radioactivity, i.e., were synthesized from the labeled glucose. Those amino acids which were not manufactured from the glucose were categorized as essential.     

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.     

Allocation of nuptial gifts in tree crickets changes with both male and female diet

Male tree crickets, Oecanthus nigricornis, offer a nuptial gift to females during mating in the form of a secretion from a dorsal metanotal gland. I examined the effects of male and female nutrient limitation on allocation of the gift. Males were fed ¹⁴C radiolabeled amino acids, placed onto high- or low-quality diets and then mated with females also maintained on high- or low-quality diets. Female acquisition of radiolabel increased, and residual radiolabel within the metanotal gland decreased, over time verifying that gift size correlates with duration of courtship feeding. High-diet females produced greater egg numbers and allocated proportionally more of the male-derived amino acids to their ovaries. As predicted, duration of courtship feeding was greater for males on high-quality diets and females on low-quality diets. Amino acid transfer, measured as the proportion of total radiolabel transferred to the female, showed a significant interaction between male and female diet. High-diet females acquired available radiolabel more rapidly from high-diet males, but low-diet females acquired available radiolabel more rapidly from low-diet males. The causes of these differences between feeding duration and nutrient allocation are discussed.     

Osmoregulation of the grass shrimp Palaemonetes pugio exposed to polychlorinated biphenyls (PCBs). I. Effecton chloride and osmotic concentrations and chloride-and water-exchange kinetics

Grass shrimp, Palaemonetes pugio, were capable of hypo- and hyper-osmotic regulation of body fluids. Hemolymph chloride and osmotic concentrations were maintained at relatively stable levels over a wide salinity range. Following an abrupt transfer from intermediate (14 and 17) to high (31 and 35) or low (1 and 2) salinities, hemolymph chloride levels exhibited initial overshoot and undershoot, respectively, of new steady-state levels. Osmotic concentrations exhibited an initial undershoot at low, but not overshoot at high salinity. Chloride space in salinity-acclimated shrimp was relatively stable at salinities from 1 to 35. Changes in chloride space following salinity transfer paralleled those of hemolymph chloride levels, and are discussed in the light of alterations in intracellular sodium concentrations reported earlier. Rate constants for chloride turnover indicated independent exchanges of sodium and chloride ions. Water-turnover measurements showed that permeability of P. pugio was greatest at the isosmotic salinity (17) and reduced at salinities which were associated with active osmoregulation. Exposure to sublethal and 96-h LC₅₀ levels of Aroclor^® 1254 did not seriously alter hemolymph chloride and osmotic concentrations, chloride space or chloride-exchange kinetics in adult shrimp. Disruption of hemolymph chloride regulation in juvenile shrimp was associated with large mortalities not observed in adults. Shrimp exposed to Aroclor 1254 at 17 S exhibited reduced water permeability similar to levels previously observed in controls at high and low salinities in response to osmotic or ionic gradients. Exposure to PCBs did not result in further reduction in permeability at the latter salinities.     

Studies on the nutritional requirements of the larval stages of Penaeus japonicus using microencapsulated diets

Fatty acid biosynthesis in the larval stages of Penaeus japonicus Bate was examined by feeding microencapsulated diets containing (1-¹⁴C) palmitic acid, and fat-free diets supplemented with defined fatty acids. Highest larval growth rates were achieved on diets containing Tapes philippinarum lipid and, when defined fatty acids were substituted, on diets containing 20:63 fatty acid. The radioactive tracer experiments indicate that 16:17, 18:0 and 18:19 fatty acids may be synthesised from palmitic acid and that P. japonicus larvae may possess the ability to elongate 18:33 to 20:53 and 22:63, and 18:26 to 20:46. However, the rates of these reactions appear to be too slow to meet the larval requirements for essential fatty acids and the 3 series of polyunsaturated fatty acids must be provided in the diet.     

The role of chemosensory behavior of Symbiodinium microadriaticum,intermediate hosts,and host behavior in the infection of coelenterates and molluscs with zooxanthellae

Symbiotic dinoflagellates, Symbiodinium microadriaticum (=zooxanthellae), may gain access to aposymbiotic hosts (i.e., those lacking zooxanthellae) by chemosensory attraction of the motile algae by the potential host or via an intermediate host. Laboratory experiments showed that motile zooxanthellae were attracted to intact aposymbiotic host animals, but not to starved symbiotic hosts. Fed symbiotic hosts and brine shrimp (Artemia sp.) nauplii also attracted motile zooxanthellae. The attraction of these zooxanthellae was directly correlated with nitrogen levels in the seawater surrounding the hosts; thus ammonia and possibly nitrate could be atractants. Brine shrimp nauplii, acting as intermediate hosts actively ingested both motile and non-motile zooxanthellae. the ingested zooxanthellae tended to remain morphologically unaltered during and after passage through the gut of the brine shrimp. Capture and ingestion of brine shrimp containing zooxanthellae by aposymbiotic scyphistomae of the jellyfish Cassiopeia xamachana led to infection of the scyphistomae with zooxanthellae. Zooxanthellae isolated from 17 different species of coelenterates and molluscs could be transferred via brine shrimp to the endodermal cells of the scyphistomae. However only 10 of these isolates persisted to establish a permanent association with C. xamachana. Scyphistomae in suspensions of motile zooxanthellae responded by a classical coelenterate feeding response, which may facilitate ingestion of the potential symbionts and establishment of a symbiosis.     

Studies on the nutrition of marine flatfish. The effect of oral administration of diethylstilboestrol and cyproheptadine on the growth of Pleuronectes platessa

Two organic compounds, diethylstilboestrol a synthetic oestrogen substitute, and cyproheptadine which acts as an appetite stimulant in some mammals, were added separately at 3 different concentrations to diets fed to plaice (Pleuronectes platessa), and their effect on growth examined. Over a 10 week period, low orally-administered diethylstilboestrol (600 g/kg moist diet) gave rise to increased weight gain, food consumption and food conversion. Higher dietary concentrations of diethylstilboestrol (up to 2400 g/kg diet) were without effect on these parameters. Cyproheptadine did not stimulate appetite, weight gain or food conversion at any of the levels tested.With an Appendix on Statistics by J. A. Pope and A. M. Shanks     

The distribution,metabolism and toxicity of 14c-DDT in model aquarium tanks with fish and sediment simulating a tropical marine environment

Studies conducted on the distribution, fate and metabolism of DDT in a model ecosystem simulating a tropical marine environment of fish, Gobious nudiceps, Lethrinus harak, Gobious keinesis, Gobious nebulosis and white shrimp (Panaeus setiferus), show that DDT concentration in the water decreases rapidly within the first 24?h. Rapid accumulation of the pesticide in the biota also reaches a maximum level in 24?h before gradually declining. The bioaccumulation factors calculated for the fish species (G. keinesis) and white shrimp (P. Setiferus) were 270 and 351, respectively, after 24?h. There was a steady build up of DDT residues in the sediment during the first 24?h which continued to a maximum concentration of 6.66?ng/g in the seawater/fish/sediment ecosystem after 3 weeks and 5.27?ng/g in the seawater/shrimps/sediment ecosystem after 2.7 days. The depuration of the accumulated pesticide was slow with only 54% lost in G. nudiceps within 3 days of exposure in fresh sea water. By contrast, depuration was fast in the white shrimp, which lost 97% of the accumulated pesticide under the same conditions. DDT was found to be toxic to two of the fish species (G. nebulosis and L. harak) and to white shrimp, and the degree of toxicity was dependent on the particular species. The 24?h LC₅₀ at room temperature for the fish species G. nebulosis and white shrimp was found to be 0.011 and 0.116?mg/kg, respectively. These levels are comparable to the ones recorded for the temperate organisms. Degradation of DDT to its primary metabolites, DDE and DDD, was found in all the compartments of the ecosystem with DDE being the major metabolite in the fish, shrimps and sediment, while in seawater, DDD dominated as the major metabolite.     

Acceptability and digestion of diets fed to larval stages ofHomarus gammarus and the role of dietary conditioning behaviour

Whilst the role of chemoreception in juvenile crustacean feeding behaviour has received considerable attention, the chemosensory responses of larval planktonic crustacean stages have been neglected. The present study investigated the acceptability of processed, microparticulate and microencapsulated diets and the possible role of chemoattractants for larval and juvenile stages ofHomarus gammarus (Linnaeus) and juvenile stages ofPalaemon elegans (Rathke). Larval and juvenile lobster were obtained from MAFF (Ministry of Agriculture, Fisheries and Food, Conwy, UK), and juvenileP. elegans were collected from the Menai Straits (Gwynedd, UK) during the summer months of 1985/1986. During the larval stages, food capture results from chance encounter, and the role of low molecular-weight chemoattractants appears to be in the determination of the edible from the inedible. Dietary conditioning behaviour in decapod larvae, which has until now not been investigated, enables the larvae to discriminate more rapidly between particles of different digestibilities. The digestibility and assimilation efficiency displayed on artificial diets in the present study suggest that lobster larvae possess limited enzymatic capabilities, although later stages are more efficient and display longer food-retention time. Growth (0.114 mm d^–1) and survival (80.0%) of lobster larvae fed natural diets were significantly better (F=14.8;P=0.001, andF=54.0,P=0.001, respectively) than of individuals fed artificial diets. Although pre-digested artificial diets (ingredients pre-treated with pancreatin) resulted in higher survival (74 and 63%) compared to microencapsulated diets (37%), there was no significant growth difference between the artificial diets tested.