The ecological significance of lipid/fatty acid synthesis in developing eggs and newly hatched larvae of Pacific cod (Gadus macrocephalus)

The lipid/fatty acid composition of marine fish eggs and larvae is linked with buoyancy regulation, but our understanding of such processes is largely restricted to species with pelagic eggs. In this study, we examined developmental changes in the lipid/fatty acids of eggs and embryos of Pacific cod (Gadus macrocephalus), a species that spawns demersal eggs along coastal shelf edges, but as larvae must make a rapid transition to the upper reaches of the water column. Adult Pacific cod were collected in the Gulf of Alaska during the spawning season and eggs of two females were artificially fertilized with sperm from three males for each female. The eggs were subsequently reared in the laboratory to determine (1) how lipids/fatty acids were catabolized during egg and larval development, and (2) whether lipid/fatty acid catabolism had measurable effects on egg/embryo density. Eggs incubated at 4°C began hatching after 3-weeks and continued to hatch over a 10-day period, during which there was a distinct shift in lipid classes (phospholipids (PL), triacyglycerols (TAG), and sterols (ST)) and essential fatty acids (EFAs: 22:6n-3 (DHA), 20:5n-3 (EPA), and 20:4n-6 (AA)). In the egg stage, total lipid content steadily decreased during the first 60% of development, but just prior to hatch we observed an unexpected 2–3-fold lipid increase (~6–9 μg individual⁻¹) and a significant drop in egg density. The increase in lipids was largely driven by PL, with evidence of long-chained fatty acid synthesis. Late-hatching larvae had progressively decreasing lipid and fatty acid reserves, suggesting a shift from lipogenesis to lipid catabolism with continued larval development. Egg density measures suggest that lipid/fatty acid composition is linked to buoyancy regulation as larvae shift from a demersal to a pelagic existence following hatch. The biochemical pathway by which Pacific cod are apparently able to synthesize EFAs is unknown, therefore representing a remarkable finding meriting further investigation.     

Changes in lipid content,lipid class composition and fatty acid composition of developing eggs and unfed larvae of cod (Gadus morhua)

Changes in the lipid class and fatty acid composition of developing eggs and unfed larvae of cod (Gadus morhua L.) were studied with the objective of determining probable requirements of cod larvae for dietary lipid. The eggs were collected on 24 March 1985 from holding tanks containing cod which had been caught off the northwest coast of Scotland. Phosphatidylcholine (PC) was the only lipid class to decline in absolute terms during embryogenesis. Catabolism of neutral lipid was initiated during the first week after hatching, and the rate of neutral lipid utilisation increased after the larvae had completely absorbed their yolk sacs. The quantity of triacylglycerol (TAG) remained constant during embryogenesis, but the percentage of 22:6(n-3) int TAG increased substantially during this period. It was calculated that ca. 33% of the 22:6(m-3) released during the process of PC catabolism was incorporated into TAG and sterol ester. The results suggest that PC, replete in appropriate essential fatty acids, should represent a major proportion of the lipid in artificial diets for fish and crustacean larvae.     

Fatty acid composition as an indicator of food intake in cod larvae Gadus morhua from Lofoten,Northern Norway

Fatty acid analyses were used to study the transition from endogenous to exogenous feeding in Arcto-Norwegian cod larvae Gadus morhua L. sampled from Lofoten waters, Northern Norway in April 1985. Fatty acids of total lipids were analysed from phytoplankton, eggs and nauplii of Calanus finmarchicus, and cod eggs and larvae. Gas chromatographic and gas chromatographic-mass spectrometric methods were used. On the basis of these analyses it is suggested that lipids of phytoplankton origin form an important part of the diet of cod larvae during the first feeding period.     

Lipids in atlantic halibut (Hippoglossus hippoglossus) eggs from planktonic samples in Northern Norway

Fertilized Atlantic halibut (Hippoglossus hippoglossus L.) eggs in different developmental stages (Days 0 to 18) were sampled from plankton in North Norway in February 1986 and analysed for lipid classes and fatty acid content. In unfertilized ovulated eggs taken from ripe fish caught in 1983/1984, polar and neutral lipids comprised ca. 71 and 30% of the total lipids, respectively, decreasing and increasing to 67 and 33%, respectively, in Stage III (11 to 18 d old) fertilized eggs. Of the polar lipids, phosphatidylcholine decreased markedly from 62% in unfertilized ovulated eggs to ca. 40% in Stage I (0 to 3 d old) fertilized eggs, while phosphatidylethanolamine increased from ca. 7 to 33%. Triacylglycerols, the major neutral lipids, increased from ca. 13% in unfertilized ovulated eggs to 16% in Stage III fertilized eggs. The total lipid in Stage I fertilized eggs had relatively low levels of polyunsaturated fatty acids (PUFA), with (n-3) PUFA accounting for only ca. 25% of the total fatty acids. The (n-3) PUFA increased to ca. 40% of the total fatty acids in Stage III, while the (n-3): (n-6) ratio increased from 4.1 to 7.0.     

Role of temperature on lipid/fatty acid composition in Pacific cod (Gadus macrocephalus) eggs and unfed larvae

During early development, oviparous fish species must use finite lipid and fatty acid (FA) reserves for both catabolism and structural components. In cold environments, developing fish have the additional constraint of maintaining membrane fluidity for metabolic efficiency (homeoviscous adaptation), resulting in further demand on lower melting point FAs like n-3 polyunsaturated fatty acids (PUFAs). To examine whether marine fish embryos physiologically adapt to changing temperature environments, we incubated Pacific cod (Gadus macrocephalus) eggs at 5 temperatures (0, 2, 4, 6, and 8 °C) in the laboratory and sampled them repeatedly during development to measure changes in lipid/FA composition. Pacific cod embryos increased n-3 PUFA content during the egg stage in all temperature treatments, with the possible exception of 0 °C, where poor survival and hatch success limited our ability for continued sampling. At the beginning of the hatch cycle, free-swimming embryos shifted from lipogenesis to lipid catabolism. The rates of lipogenesis and catabolism were temperature dependent, and the distinct increase in unsaturated fatty acids at temperatures <8 °C was consistent with homeoviscous adaptation theory. However, with the possible exception of embryos at 0 °C, the relative amounts of essential fatty acids (e.g., EPA, DHA, AA) were conserved in a similar manner across incubation temperatures. Collectively, these data suggest Pacific cod are capable of homeoviscous adaptation but cannot tolerate temperatures approaching 0 °C despite their possible ability to biosynthesize PUFAs from other energetic sources.     

Physiological energetics of developing embryos and yolk-sac larvae of Atlantic cod (Gadus morhua). II. Lipid metabolism and enthalpy balance

This paper presents quantitative data for the changes in the contents of total lipids, lipid classes and their associated fatty acids, together with the changes in caloric contents of developing eggs and yolk-sac larvae of Atlantic cod (Gadus morhua L.). During development between Day 0 to 28 post fertilisation, 32% of catabolic metabolism was fuelled by lipids. On a mass-specific basis, polar lipids (mainly phosphatidyl choline) contributed 60%, and neutral lipids (mainly triacyl-glycerol) contributed 40% to this catabolic component, with each class supplying similar amounts of fatty acids as fuel. The fatty acids supplied by phosphatidyl choline were catabolised non-selectively (i.e. in proportion to their presence in the egg), with about half of them being polyunsaturated. However, of the fatty acids esterified in triacylglycerol, the larvae showed an apparent oxidation preference for monoenes over polyunsaturates or saturates. Routine rates of oxygen consumption and ammonia production were related to the caloric contents of the eggs and larvae in order to derive an enthalpy balance equation (of the form P=C+R+E) for an Atlantic cod larva during its period of endogenous nutrition. For the interval of Day 0 to 25 post fertilisation (the period of yolk dependence), integration of the physiological and caloric data revealed that Atlantic cod larvae conserved 53% of yolk enthalpy (C) for growth (P), 42% was dissipated due to metabolism (R) while only 5% was lost via excretion (E).     

Discrimination between two reared stocks of cod (Gadus morhua) from the Faroe Islands by chemometry of the fatty acid composition in the heart tissue

The composition of fatty acids in total lipids and in phosphatidylcholine and phosphatidylethanolamine in the heart tissue from two reared stocks of cod (Gadus morhua L.) was determined by a chemometric method, consisting of methanolysis, gas chromatography of the resulting fatty acid methyl esters and multivariate statistical treatment, by principal component analysis, of the analytical data. The two reared stocks of cod from the Faroe Bank and the Faroe Plateau had significantly different fatty acid profiles in all three groups of lipids. This difference is expected to be purely genetic and free of biotic and abiotic impacts on the fatty acid profiles. The observed clear-cut distinction suggests that the method may have the potential to discriminate between the corresponding wild stocks, although possible variation in the fatty acid profile caused by internal and environmental factors must be better understood. Received: 6 August 1999 / Accepted: 6 January 2000     

Changes in biochemical and fatty acid composition of the razor clam <Emphasis Type="Italic">Solen marginatus</Emphasis> (Solenidae: Bivalvia) during larval development

Aquaculture studies have revealed that polyunsaturated fatty acids are critical for maintaining substantial growth, survival and reproductive rates, and high food conversion efficiencies for a wide variety of marine and freshwater organisms. The aim of this study was to investigate the gross biochemical and fatty acid composition of both neutral and polar lipid compartments of the razor clam Solen marginatus throughout embryonic and larval development. High levels of stored reserves in S. marginatus eggs allow a short larval development, lasting only 8 days. The energy required for embryogenesis was obtained from stored proteins. During larval development from D-shaped veliger until settlement, protein, lipid, and carbohydrate reserves were indistinctly stored for metamorphosis. Although total lipids increased, fatty acids in both neutral and polar lipids decreased during embryonic development. The depots allow a short larval development in which settlement is reached with lower amounts of stored neutral and polar lipids than the contents found in the oocytes. Non-methylene-interrupted dienoic fatty acid levels were similar to those of some polyunsaturated fatty acids, with increasing percentages at the onset of metamorphosis. This study indicates that S. marginatus exhibits a different pattern in the use of gross biochemical and fatty acid reserves during larval development compared to other razor clam and bivalve species, mainly due to the large size of its eggs and the short larval development stage reported in this species.     

Fatty-acid composition of ovulated eggs from wild and cultured turbot (Scophthalmus maximus) in relation to yolk and oil globule lipids

The fatty-acid composition of lipids from ovulated eggs of wild and cultured turbot was investigated in order to estimate the nutritional requirements during embryonic and early larval development. Lipid comprised 13.8±0.5% (n=5) and 13.2±0.7% (n=7) of the egg dry weight in wild and cultured turbot, respectively. Polyunsaturated fatty acids (PUFA) of the (n-3) series accounted for 39% of total fatty acids in total lipid of both wild and cultured fish. The predominant (n-3) FUFA was docosahexaenoic acid (22:6 n-3), which also was the most abundant fatty acid in turbot eggs and comprised 24 and 23% of the total egg fatty acids in wild and cultured fish, respectively. Phospholipids, triacylglycerols and cholesterol-wax esters of turbot eggs all exhibited a specific fatty-acid profile distinctly different from that of total lipid. The general pattern of the fatty-acid distribution in lipids of eggs from wild and cultured turbot was similar, but the relative amount of 18:2(n-6) was considerably higher and 20:1(n-9) slightly higher in cultured fish. These differences were extended to all lipid classes and probably reflect the dietary intake of certain vegetable and marine fish oils. Calculations based on light microscopical studies showed that 55 to 60% of the total lipids in cultured turbot eggs are confined to the oil globule. The size of the oil globule remained constant during embryogenesis, and a reduction in size occurred first after hatching and mainly after yolk depletion. This implies that the total amount of lipids utilised during the embryonic development is considerably less than the total lipids present in ovulated turbot eggs. Comparison of the fatty-acid composition of total lipids from eggs and vitellogenin of wild turbot reveals that egg lipids contained a lower level of saturated and a higher level of monounsaturated fatty acids. Eggs also contained wax esters, which were not detected in vitellogenin, suggesting that vitellogenin is not the only source of lipids for turbot eggs.     

Studies on the nutrition of marine flatfish. The effect of dietary fatty acids on the tissue fatty-acids of the plaice Pleuronectes platessa

The ratios of polyunsaturated fatty acids in the lipids of hatchery-reared plaice [Pleuronectes platessa (L.)] maintained on a diet containing corn oil and freeze-dried cod muscle accurately reflect ratios of polyunsaturated fatty acids in the diet. Such plaice have much greater levels of triglycerides in both their livers and extrahepatic tissues than wild plaice of comparable size; the total amount of 22:6 3 acid in triglycerides of the two groups is, however, very similar. Transfer of experimental plaice to a fat-free diet caused a marked depletion of triglycerides from both livers and extrahepatic tissues with almost all fatty acids being reduced in amount. Addition of 12:0 and 14:0 ethyl esters (4% of each) to the fat-free diet caused some accumulation of triglycerides in extrahepatic tissues. Further addition of 18:2 6 and 18:3 3 acids (both at levels of 0.4%) to the diet caused an accumulation of triglycerides in the liver. These triglycerides had increased levels of saturated and monounsaturated fatty acids up to C 18 but the levels of higher polyunsaturated fatty acids were not inereased. There was relatively little change in the fatty acids of phospholipids throughout these experimental dietary regimes. The levels of phospholipids were constant and very similar to those of wild plaice. The results indicate: (1) plaice fed a diet relatively low in 22:6 3 acid may accumulate relatively large amounts of triglyceride in order to store adequate amounts of 22:6 3 acid; (2) plaice are capable of synthesising saturated and mono-unsaturated fatty acids up to C 18 in the presence of dietary 12:0 and 14:0 acids; (3) 18:2 6 and 18:3 3 acids in the diet do not undergo chain elongation and further desaturation in plaice with the dietary regimes we used.     

Effect of lipid emulsions on production and fatty acid composition of eggs of the scallop<Emphasis Type="Italic"> Argopecten purpuratus</Emphasis>

The impact of supplementing lipid emulsions rich in eicosapentaenoic acid (EmEPA), docosahexaenoic acid (EmDHA) or saturated fatty acids (EmCOCO) to a standard algal diet [3:1 mixture of Isochrysis galbana (T-iso) and Chaetoceros neogracile, St-diet] on Argopecten purpuratus broodstock was evaluated. Broodstock fecundity was compared as well as the egg quality in terms of lipid content, fatty acid composition and lipid class distribution. Fecundity was defined as the number of eggs released in the spawning process, since spawning was virtually complete. Results indicated that the total lipid content of the eggs of A. purpuratus was diet independent. A greater energy reserve was spent on a larger number of oocytes and not on bigger sized oocytes with a higher lipid content. The lipids supplied through the emulsions were at least partially allocated to the eggs, demonstrating that the fatty acid composition of the eggs could be manipulated, especially the neutral lipid fraction. Levels of EPA changed more rapidly than DHA levels, supporting the observation that they fulfilled an energetic and structural role, respectively. The St-diet supplemented with 50%EmCOCO resulted in a significantly higher fecundity compared to the algal diet supplemented with 25%EmEPA+25%EmDHA and the non-supplemented algal diet. It would seem that saturated fatty acids (SAFA) were more easily or preferentially incorporated in the female gonads of A. purpuratus. The relative content of SAFA and 18:2( n-6) in these eggs rose significantly. The relative content of the highly unsaturated fatty acids, EPA and DHA, on the other hand was substantially lower in the neutral lipid fraction, but hardly affected in the polar lipid fraction. It appeared that the maintenance of an adequate DHA/EPA ratio (approximately 1.2) was more important than the absolute levels of the two fatty acids, as long as a threshold value was reached.     

Water and salt balance of halibut eggs and larvae (Hippoglossus hippoglossus)

Eggs of halibut [Hippoglossus hippoglossus (L.)] have a negative buoyancy in sea water of 35 S, in contrast to eggs of most flatfish species. The cause of this was investigated. The osmolality of the yolk is 350–420 mOsm during embryonic development. This is within the range for marine teleost serum and for yolk of pelagic eggs. Concentrations of major inorganic ions are comparable with those of pelagic plaice eggs [Pleuronectes platessa (L.)]. The values for Na⁺, K⁺ and Cl^- are 6, 85, and 64 mmol · (1 H₂O)^-1 after fertilization, and at the time of hatching the corresponding values are 17, 11, and 80. Large amounts of other inorganic constitutents are excluded for osmotic reasons. Malfunction in the regulation of osmolality or of inorganic constituents is thus unlikely to be the cause of negative buoyancy. The relative dry weight of the chorion (egg shell) in halibut eggs is less than in several pelagic egg types, excluding the chorion as the main contributor of negative buoyancy. It is concluded that a high content of organic matter in the rest of the egg is the cause of the negative buoyancy.     

Decreased lipid storage in juvenile Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>) during settlement in cold-water eelgrass habitat

We characterized the prey field and the lipid classes/fatty acids in the flesh of age 0 juvenile cod (Gadus morhua) during their late-summer/fall arrival and settlement into eelgrass (Zostera marina) in coastal Newfoundland. Examination of available prey demonstrated a high abundance of small zooplankton (Acartia, Microsetella and Oithona sp.) with no larger Calanus sp. prey. Breakpoint analysis showed significant changes in the accumulation of relative (mg g⁻¹ wet weight) and absolute (μg fish⁻¹) amounts of lipid with standard length at the time of settlement (~60 mm standard length). Settling juvenile cod showed an alternate lipid utilization strategy where they catabolized phospholipids (PL) to a greater extent than triacylgylcerols (TAG). Polyunsaturated fatty acids (PUFA) content in cod flesh decreased as fish grew indicating that nearshore zooplankton quality was not optimal for PL formation. The dramatic reduction in cod PL was likely due to both catabolism of muscle and a lack of dietary PUFA suitable for PL synthesis. However, juvenile cod continued to grow, leading to decreased lipid stores and suggesting that cod settling into eelgrass are under intense selection pressure for growth prior to the onset of winter, possibly as a means of escaping gape-limited predation. These data contrast better-studied freshwater and estuarine systems in which lipid storage is critical for successful overwintering.     

Egg buoyancy of Baltic cod (Gadus morhua) and its implications for cod stock fluctuations in the Baltic

Successful spawning of Baltic cod (Gadus morhua L.) is restricted to the deep basins of the Baltic Sea and is dependent on abiotic conditions such as salinity and water oxygen content. Due to irregular inflows of saline water from the North Sea, the conditions for spawning fluctuate and consequently so does the cod stock. In May and June 1990 eggs were obtained from spawning cod caught off northern Gotland, Sweden. Our investigation revealed neutral egg buoyancy for Baltic cod at a salinity of 14.4±1.1 ppt, with a slight decrease some days before hatching. Today salinities of this magnitude occur only in the Bornholm basin, whereas in the Gdansk and Gotland basins the eggs sink toward the bottom and are exposed to lethal oxygen conditions.     

Lipid composition of eggs and adductor muscle in giant scallops (Placopecten magellanicus) from different habitats

Scallops (Placopecten magellanicus Gmelin) were collected during August 1989 from shallow water (10 m) and deep water (31 m) habitats at Sunnyside, Trinity Bay, Newfoundland, to compare the lipid composition of eggs and adductor muscle tissue. Less favorable food levels and lower temperature conditions associated with deeper water have previously been shown to produce slower growth and reduced fecundity in individuals from this habitat. Triacylglycerol reserves consistently accounted for 60% of the total lipids present in both groups. The total lipid content of the eggs and the composition of their triacylglycerol fatty acid pools were similar in shallow water and deep water scallops, indicating very little if any nutritional difference between the two groups. Relative to their counterparts from shallow water, individuals from deeper water contained higher proportions of docosahexaenoic acid [22:6(n-3)] in the egg phospholipids and higher levels of 24-methylenecholesterol (a phytosterol commonly found in diatoms) in the adductor muscle. Differences in fatty acid composition are interpreted as biochemical adjustments of cell membranes to increase membrane fluidity, thereby compensating for the lower temperatures prevailing at the greater depth.     

Lipid storage and composition in tropical surgeonfishes (Teleostei: Acanthuridae)

The lipid composition of tropical marine reef fishes is poorly known, despite their use as food by local human populations and recent interest in health-related benefits of fish lipids. We examined the composition of lipids from epaxial muscle, liver, and two storage sites [mesenteries surrounding the gut (intraperitoneal fat, IPFs) and retroperitoneal fat bodies (FBs) posterior to the peritoneal cavity] in three species of surgeonfishes from Ishigaki Island, Japan: Naso lituratus (Bloch and Schneider, 1801), Acanthurus lineatus (Linnaeus, 1758), and A. bariene (Lesson, 1830). Triacylglycerols dominated all samples of neutral lipid and constituted ≥ 99% of FBs and IPFs. Polar lipids generally contained large fractions of phosphatidylethanolamine and phosphatidylcholine. Quantified fatty acids ranged in length from C₁₄ to C₂₄. C₁₆ fatty acids prevailed (>35% of neutral fatty acids, >23% of polar fatty acids), although C₁₈ (>16 and >14%, respectively) and C₂₀ acids (>8 and >19%, respectively) were also common. Saturated fatty acids, dominated by palmitic acid (16:0), comprised 38.7 to 50.7% of acids from neutral lipids and 30.8 to 41.1% from polar lipids. The most common monounsaturated acids were 18:1n9 and 20:1n9. Polyunsaturated acids were prevalent in polar lipids (especially 20:4n6, 20:5n3, 22:2n3, 22:5n3, 22:5n6 and 22:6n3). Common polyunsaturated acids of neutral lipids were 18:2n6, 18:4n3, several n-3 and n-6 C₂₀ acids, 22:2n3 and 22:5n3. IPF and FB were almost identical across species, and lipids of fat bodies (IPFs, FBs) were more similar to those of muscle than those of liver for all three species. The FBs appear to constitute an accessory storage site, which overcomes constraints on lipid storage imposed by a small, inflexible abdominal cavity that contains both viscera and consistently voluminous gut contents. Fatty acid signatures indicate that largely overlooked epiphytic or epilithic diatoms contribute significantly to lipid acquisition. The combination of large quantities of both saturated and n-3 and other polyunsaturated fatty acids in surgeonfishes, in contrast to low saturates and high polyunsaturated acids in lipids of commercially important cool-water fishes, suggests that a study of dietary effects of fish lipids on human inhabitants of the tropics may be instructive insofar as human health and nutrition are concerned. Received: 16 March 1998 / Accepted: 6 August 1998     

Maternal allocation of lipid classes and fatty acids with seasonal egg production in Atlantic cod (Gadus morhua) of wild origin

A suite of characteristics is often used to assess egg quality as these properties potentially play important roles in progeny survival and growth. Our objective was to assess egg characteristics including lipid biocomposition of an iteroparous, batch-spawning teleost of wild origin. Maternal allocation to egg number was generally dome-shaped (5 of 8 females) and egg size declined over the breeding season for eight breeding pairs of wild Atlantic cod (Gadus morhua) (n = 43 batches). Egg lipid composition ranged considerably among females and between egg batches within females (e.g., phospholipids 40–86 %; polar 47–87 % and neutral lipids 15–52 % of total lipids; polyunsaturated fatty acids 16–50 % of total fatty acids). Principal component analyses revealed significant inter-relationships among maternal traits, batch sequence and fecundity, and egg size and composition. Seasonal trends with regard to lipid deposition were variable; three females showed consistent declines in lipid parameters (μg egg⁻¹) with both batch number and egg diameter, one female showed consistent increase and the four remaining females showed no trend. The three females that exhibited seasonal declines in egg lipid content were characterized as having high fertilization success (>75 %). Our findings highlight the variability in lipid allocation to eggs of batch spawners of wild origin and characterize the composition of endogenous reserves available during embryogenesis and yolk sac larval stages.     

Fecundity,brood loss and egg development through embryogenesis of <Emphasis Type="Italic">Armases cinereum</Emphasis> (Decapoda: Grapsidae)

The present work is a comprehensive study of reproduction and embryonic development of Armases cinereum. Ovigerous A. cinereum (Bosc, 1802) females from Sebastian Inlet, Florida (9.88–19.4 mm CW) lay 2,000–12,000 eggs per brood, depending on their CW (mm): fecundity = 24.662 CW^1.9432. A. cinereum displayed significant brood loss through development (ca. 500 eggs per brood) independently from their CW (no senescence). However, since smaller females lay fewer eggs than larger ones, the percentage of eggs lost during embryonic development is greater in smaller females. The number of eggs carried on a later stage of development (potential fertility = 5.5593 CW^2.4417) is a more accurate estimate of the reproductive output and subsequent recruitment. Egg volume increased during development (64%, 0.025–0.041 mm³ or 0.36–0.43 mm of diameter, N = 270) and was strongly correlated with egg water content increase (19.21%, r = 0.89). Lipids, particularly fatty acids, seem to be the major energy source for embryonic development, decreasing 56.31 and 37.08% (respectively) during embryonic development; both are negatively correlated with egg volume (r = −0.90). The utilization of fatty acids through the different developmental stages of A. cinereum is presented. The most consumed fatty acids are the monounsatured (43.33 μg mg⁻¹ dw), followed by the saturated (29.91 μg mg⁻¹ dw) and polyunsaturated (24.03 μg mg⁻¹). Palmitic (16:0) and linoleic (18:2n-6) acids are preferentially consumed (19.5 and 17.9 μg mg⁻¹ dw, respectively). The high proportion of essential polyunsaturated fatty acids of C₁₈ and C₂₀ reflects the consumption of primary producers such as mangrove leaves. EPA/DHA ratio (2.85–3.84) and low DHA content indicated that this species appears in a medium-low level of the trophic chain. The low ratio of 18:1n-7/18:1n-9 and high percentage of 18:1n-9 (marker of carnivory) may be a sign of the consumption of juvenile invertebrates. The high percentage of odd-numbered FA indicated the occurrence of detritivores/scavenger behaviours. The fatty acid composition of the eggs reflects adult feeding ecology (omnivorous) and habitat.     

Lipid biochemistry and reproductive biology in two species of Gammaridae (Crustacea: Amphipoda)

Gammarus oceanicus Segerstråle, 1947 and Echinogammarus marinus (Leach, 1815) were sampled during the breeding season from Oslofjord in 1984, and their lipid composition examined in relation to reproductive condition. In G. oceanicus, female lipid content increased as the ovary matured. Both the amount of lipid stored and the rate of accumulation were greater in spring than in winter. Spring eggs contained 12.4 g lipid, of which 63% was triacylglycerol and 27% phospholipid. Both fractions decreased steadily during embryonic development. Winter eggs contained 19.2g lipid, of which 52% was triacylglycerol and 43% phospholipid. During the early stages of embryonic development the amount of phospholipid decreased sharply, whereas that of triacylglycerol increased, suggesting that some of the fatty acid released from phospholipid was sequestered temporarily as triacylglycerol. When newly spawned, both winter and spring eggs were richer in monoenoic fatty acids than adult amphipods and these acids were the major fuel consumed during development. 6 fatty acids were utilised more slowly than 3 acids, and egg carotenoid pigment content remained constant. Female E. marinus increased in lipid content as the ovary matured. Spring eggs contained 14.7 g lipid when newly spawned and this increased to 16.6 g during the early stages of development. This increase was entirely triacylglycerol, which declined in later stages; the source of the extra lipid was unclear. Eggs contained very little phospholipid or sterol, and both of these components remained at a steady low level during development. E. marinus eggs were not significantly rich in thonoenoic acids compared with adults, and saturated, monoenoic and polyenoic acids were utilised about equally during development. Both adults and eggs were rich in 20.46, which was utilised at a slower rate than the 3 polyunsaturated acids during embryonic development; again, egg carotenoid pigment content remained constant. In both species there was a decrease in the size of the egg (and as a result, of the newly hatched juvenile), but an increase in total reproductive output (i.e., the total weight of the egg clutch) per female as the breeding season proceeded. The reproductive output of an individual female is probably related to food availability during the period of ovarian maturation, whereas the size of an individual egg is dictated largely by feeding conditions for the juveniles once they are independent of the female. The different patterns of lipid utilisation during development found in this study emphasize the flexibility of response in the reproductive biology of gammarid amphipods. It is not yet possible, however, to relate the differing patterns in a simple way either to egg size or total female reproductive output. Two outstanding problems are the source of extra triacylglycerol during the early stages of development of E. marinus and the metabolic cost of brooding eggs.     

Energy substrates for eggs and prefeeding larvae of the dolphinCoryphaena hippurus

Changes in the chemical composition of developing dolphin (Coryphaena hippurus) eggs and prefeeding yolksac larvae were determined in order to estimate probable dietary requirements of first-feeding larvae. Daily dry matter, protein nitrogen (PN), non-protein nitrogen (NPN), lipid, gross energy content, fatty acid and amino acid profiles from Day 1 to Day 2 eggs and Day 1 to Day 3 larvae were compared. Lipid was the primary endogenous energy source accounting for the daily caloric deficit through both the egg and larval stages, except over the day of hatching. The catabolism of lipid by embryos (0.078 cal d^–1) was greater than that by yolksac larvae (0.036 cal d^–1). The higher demand for energy by embryos was related to a greater rate of protein synthesis during the egg stage. The ratio of PN:NPN increased during egg development without change in total nitrogen content, but was constant throughout the yolksac larvae period. The lipid content per embryo did not decrease over the hatching period (Day 2 to 3, postspawning). However, there was a loss in amino acid content not totally accounted for by sloughing of the chorion at hatching. This loss, as protein, accounted for 0.053 cal of gross energy, which represented 70% of the total estimated energy needs of the fish over this period. Loss of non-essential amino acids (25%) was higher than that of essential amino acids (13%). Proline and tyrosine accounted for 32% of the total loss of amino acids at this time. The only preferential use of fatty acids over any period was a small but significant drop in the content of C22:6n-3 prior to the onset of feeding (Day 5, postspawning). It is speculated that the pattern of energy-substrate use of first-feeding dolphin larvae will reflect the pattern of endogenous energy use during the egg and prefeeding yolksac larval stages. Diets or feeding regimens with lipid as the primary energy source, and containing a fatty acid profile similar to that of eggs or yolksac larvae, should be useful in culturing this species, at least during the early feeding stages.