Free amino acids as energy substrate in developing eggs and larvae of the cod Gadus morhua

The content of free amino acids (FAA) in the cod (Gadus morhua L.) egg is about 200 nmol at spawning, decreasing by about 100 nmol/egg during the egg stage and about 75 nmol/larva during the yolksac larval stage. Together, alanine, leucine, serine, isoleucine, lysine, and valine account for about 75% of the decrease. Ammonium accumulates gradually during the egg stage and is quickly excreted after hatching. The body protein content is maintained during the egg and yolksac larval stages. The measured oxygen uptake of the cod embryo during the egg and yolksac larval stages accounts for about 85% of the oxygen necessary to catabolize the FAA disappearing during this period. Ammonia excretion of the cod embryo, as taken from literature data, is similar to the expected ammonia production from catabolism of the FAA. Our data suggest that FAA are a major substrate for aerobic energy production in cod eggs and yolksac larvae. The implication of this finding for the production of a favourable first-feed for cod and other cultivated marine fish larvae, and for the selection of high quality eggs of marine fishes, is stressed.     

The importance of free amino acids to the energy metabolism of eggs and larvae of turbot (Scophthalmus maximus)

A study was undertaken to establish the role of free amino acids (FAA) in aerobic energy dissipation in embryos of turbot (Scophthalmus maximus) which contain an oil globule in the egg. Laboratory-reared developing eggs and larvae (15°C, 34 salinity) were measured for oxygen uptake, ammonia excretion, contents of FAA, protein, and ammonium, and volumes of yolksac and oil globule. Newly spawned eggs from different batches contained 55 to 90 nmol egg^–1 of FAA. Resorption of FAA occurred in parallel with the consumption of yolk. Resorption of the oil globule, however, occurred predominantly after hatching and mainly after yolk resorption. The combined data suggest that approximately 70% of the FAA are utilized as an energy substrate, while the rest are polymerized into body proteins. FAA become a significant energy substrate in the early egg stage and account for 100% of the aerobic energy dissipation 2 d after Fertilization then decrease to ca. 60% at the time of hatching. Lipids derived from the oil globule seem to be the main fuel after hatching and account for ca. 90% of the energy dissipation at the onset of first-feeding. Thus, the energetics of fish embryos which contain an oil globule seems to be different from those that depend exclusively on the nutritional reserves of the yolk.     

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.     

Growth, carbon, nitrogen and caloric content of Solea senegalensis (Pisces: Soleidae) from egg fertilization to metamorphosis

Eggs and larvae of the Senegal sole, Solea senegalensis Kaup, were reared from fertilization until the end of metamorphosis, which occurs by Day 17 after hatching at 19.5 °C. Changes in energy content and biomass quality were studied in terms of dry weight and of carbon, nitrogen and energy content. S. senegalensis spawned eggs of about 1 mm diameter which hatched 38 h after fertilization. Average dry weight of individual eggs was 46 μg, the chorion accounting for about 18% of total dry weight. Gross energy of recently fertilized sole eggs was approximately 1 J egg⁻¹. From fertilization to hatching, eggs lost 8% of their total energy (chorion not included). After hatching, larvae lost 14% of their initial energy until the start of feeding which occurred about 48 h afterwards. The principal components catabolized during embryogenesis were carbon-rich compounds that decreased by 26%, while nitrogen-rich compounds decreased by only 10% and were practically unaltered from hatching to the start of feeding. Feeding larvae displayed constant growth during the period studied (specific growth rate on a dry weight basis was 0.26 d⁻¹). The relative proportion of carbon and nitrogen content revealed an accumulation of high energy compounds in the days before metamorphosis. By Day 14, the energy content reached values similar to those of recently hatched embryos, but decreased again during metamorphosis. Received: 10 June 1998 / Accepted: 28 January 1999     

Energy metabolism during development of eggs and larvae of gilthead sea bream (Sparus aurata)

Developing eggs and larvae of laboratory-reared gilthead sea bream (Sparus aurata) maintained in filtered seawater (40 ppt) at 18°C, were measured for oxygen uptake, ammonia excretion, contents of free amino acids (FAA), protein, fatty acids (FA) accumulated ammonia, and volumes of yolk-sac and oil globule. Absorption of the yolk coincided with the consumption of FAA and was complete ca. 100 h post-fertilisation. Amino acids from protein were mobilised for energy in the last part of the yolk-sac stage. Absorption of the oil globule occurred primarily after hatching following yolk absorption, and correlated with catabolism of the FA neutral lipids. Overall, FAA appear to be a significant energy substrate during the egg stage (60 to 70%) while FA from neutral lipids derived from the oil globule are the main metabolic fuel after hatching (80 to 90%).     

Lipids and fatty acids as indicators of egg condition,larval feeding and maternal effects in Cape hakes (<Emphasis Type="Italic">Merluccius paradoxus</Emphasis> and <Emphasis Type="Italic">M. capensis</Emphasis>)

Cape hakes, Merluccius paradoxus and M. capensis, are important gadoid fish that are commercially harvested in the Benguela Current system off Namibia and South Africa. The aim of this study was to elucidate the nutritional condition and feeding preferences of their larvae. Hake eggs and larvae were sampled in austral spring of two consecutive years, 2007 and 2008, off the west coast of South Africa. They were identified to species using genetics, and total lipid content and fatty acid (FA) composition were analysed for each individual egg and larva to compare the condition of different early life stages of both hake species. Higher abundances of M. paradoxus eggs and larvae were consistently found compared to M. capensis. In both species, eggs contained wax esters (WE) and had significantly higher lipid content per dry mass than larvae. Lipid content as well as FA composition changed with the developmental stage of larvae. Quantities of essential fatty acid (EFA) increased with feeding of larvae due to dietary lipid incorporation. In 2007, yolk-sac larvae contained significantly lower total lipids than in 2008. It is argued that this was due to reduced lipid transfer by the spawning females to the eggs. These findings indicate that maternal effects are important in determining condition of hake larvae and that this may have an effect on their survival and subsequent recruitment.     

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).     

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.     

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.     

Physiological energetics of developing embryos and yolk-sac larvae of Atlantic cod (Gadus morhua). I. Respiration and nitrogen metabolism

This paper provides the basis for a general model of catabolic metabolism for developing embryos and yolk-sac larvae of Atlantic cod (Gadus morhua L.). Yolk-dependent routine rates of oxygen consumption, ammonia excretion, and accumulation of ammonium ions were related to quantitative changes in contents of glucose, glycogen, lactate, free amino acids, proteins and lipid classes (lipid classes published separately) in order to determine the rate and sequence of catabolic substrate oxidation that occurs with development at 6.0°C, 34.5 S. The stoichiometric relation of the oxygen consumption and total ammonia production to substrate utilisation indicated that during the first 2 to 3 d of development, glycogen was the sole substrate of oxidative metabolism. After formation of the syncytium, free amino acids (75%) together with polar lipids (13%, mainly phosphatidyl choline) and neutral lipids (9%, mainly triacylglycerol) comprised the metabolic fuels of embryonic development. Following hatch (Day 16 post fertilisation), the fuels were free amino acids (32%), polar lipids (20%, mainly phosphatidyl choline), neutral lipids (17%, mainly triacylglycerol) and proteins (31%). Thus, the catabolic metabolism of endogenously feeding Atlantic cod larvae was predominantly fuelled by amino acids (67%) and lipids (32%), while glycogen only accounted for 1% of the total enthalpy dissipated. It is proposed that the above sequence of catabolic substrate oxidation is also generally applicable to other cold-water fishes which spawn eggs that do not contain oil glubules.     

Lipid and protein utilisation during early development of yellowtail kingfish (<Emphasis Type="Italic">Seriola lalandi</Emphasis>)

The pelagic yellowtail kingfish Seriola lalandi has become a target species for aquaculture in Asia and Australasia. Australasian production is reliant on larviculture from eggs of captive brood stock; however, knowledge regarding the nutritional requirements of larvae of this species is still scarce, particularly in relation to lipids. As a first step in establishing these requirements, eggs and larvae from captive S. lalandi brood stock were examined for differences in total protein, total lipid and lipid classes between individual spawning events, over the spawning season, and during larval development from fertilisation to 15 days post hatch. Results indicate that total protein egg⁻¹ varied significantly between individual spawning events within a season, but neither total lipid nor total protein egg⁻¹ varied significantly across the spawning season. Brood stock egg lipids were made up of approximately 60% phospholipid, 25% wax and/or sterol esters (WE), 15% triacylglycerol (TAG), and small amounts of sterols and free fatty acids. During the early larval period, both WE and TAG were utilised concurrently for energy. The larvae experienced very high mortality around 5–7 days post hatch, which coincided with very low levels of all neutral lipid classes. Although many other factors may also influence larval mortality, these results indicate that lipid provisioning may be an important factor in larval survival during the critical period around first-feeding in this species. Examination of ratios of TAG:ST, often used as a condition index in fish larvae, suggested that some of the larvae were suffering from starvation. However, as egg-derived WE appears to provide a significant source of energy during the early larval period in S. lalandi, it is suggested that WE should be included in any index of larval nutritional state.     

Physiological and biochemical aspects of embryonic and larval development of the winter flounder Pseudopleuronectes americanus

Eggs and larvae of the winter flounder Pseudopleuronectes americanus Walbaum were hatched and raised in the laboratory under controlled conditions. Biochemical composition was measured during development and found to be similar to that of other species: 65 to 80 percent protein, 15 to 30 percent fat, and 0 to 5 percent carbohydrate. Ash content was 7 to 10 percent of dry weight. The chorion comprised more than half of the weight of an egg and the data suggested that it was possibly a source of nutrition to the developing embryo. The sequence of utilization appeared to be carbohydrate and then protein to hatching, lipid, mixed lipid and protein, the predominantly protein until feeding began. Carbohydrate was accumulated at first feeding and depleted when growth began. Protein and lipid were deposited in approximately constant proportions. Respiration rates of eggs were low, 0.002–0.015 l O₂ egg^-1 h^-1, but rose gradually from fertilization to hatching. Respiration rates of early larvae were from two to eight times that of eggs (0.033–0.131 l O₂ larva^-1 h^-1). Variation in larval respiration rates indicated a three-fold difference in rate according to level of activity. Eggs excreted ammonia at an increasing rate from fertilization to hatching. Larvae excreted ammonia, primary amines, and other unidentified organic nitrogenous substances. Rates of excretion and proportions of excretory products varied with stage of development. Primary amine excretion was variable and a major component in early stages. Ammonia-N excreted was two to 20 times primary amine N excreted. Unidentified substances were the predominant form of N excretion during early feeding. Ammonia accounted for most of the N excreted in older larvae. Early specific growth rates were 2.1 and 5.5%. Net caloric conversion and net and gross nitrogen efficiencies were low in first feeding larvae compared to adult fishes (32.2, 27.7, and 10.7% respectively).Contribution no. 5071 from the Woods Hole Oceanographic Institution     

Food availability and physiological state of sea urchin larvae (<Emphasis Type="Italic">Strongylocentrotus purpuratus</Emphasis>)

Food availability is highly variable in the ocean. Many species of marine invertebrates have a larval form that depends upon exogenous nutrients for growth, yet there are few biochemical and physiological indices for determining changes in the nutritional status of larvae. In this study, the effects of food availability on biochemical compositions and metabolic processes of larvae of the sea urchin, Strongylocentrotus purpuratus, were determined. Larvae were cultured under different food concentrations (fed-to-excess and unfed) and a suite of biological processes assayed, ranging from measurements at the level of the whole organism to that of specific molecules. These data were normalized to DNA content (an index of cell number) to allow comparisons of physiological rates in larvae of different sizes. Changes in the following were measured during larval growth: free amino acid pool, protein, lipid classes (cholesterol, free fatty acids, hydrocarbons, phospholipids, triacylglycerol), enzyme activities (Na⁺, K⁺-ATPase and citrate synthase), and respiration rates. In growing larvae, the two key components that showed differential cell-specific content relative to unfed larvae were glycine in the free amino acid pool and phospholipids. Additionally, several lipid classes were detectable only in fed larvae (cholesterols, free fatty acids, and hydrocarbons). While triacylglycerols were present in eggs and utilized during pre-feeding development, they were not re-accumulated at detectable levels in feeding larvae. Respiration rates, protein content, and enzyme activities were all similar on a cell-specific basis, showing that these variables did not provide useful indices of differences in physiological state between fed and unfed larvae. In contrast, measurements of the cell-specific content of glycine and certain lipid classes did provide useful indices of physiological state of larvae. Application of these indices could potentially allow for determinations of nutritional state of larvae in the ocean.     

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.     

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     

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.     

Maternal provisioning for larvae and larval provisioning for juveniles in the toxopneustid sea urchin <Emphasis Type="Italic">Tripneustes gratilla</Emphasis>

Lipid and protein biochemistry of eggs (84 μm in diameter), embryos and early larvae of the tropical echinoid Tripneustes gratilla (Linnaeus 1758) were quantified to determine how maternal provisions are used to fuel development of the echinopluteus. The eggs contained a mean of 30.82 ng lipid and 87.32 ng protein. Energetic lipids were the major lipid component (55.52% of total lipid) with the major class being triglyceride (TG: mean 15.9 ng, 51.58% of total). Structural lipid was dominated by phospholipid (PL: mean 11.18 ng, 36.26% of total). Early embryogenesis was not a major drain on egg energetic lipid and protein. Development of the functional feeding larva used ca. 50% of initial egg energetic lipid and most of this was TG. Maternal TG was still present in the 8-day echinoplutei and it was estimated that this energetic lipid would be depleted in unfed larvae by day 10. There was no change in PL. In a separate experiment lipid biochemistry of rudiment stage larvae and early developing juveniles were quantified to determine how lipids are used during metamorphosis. Fed larvae accumulated lipid (mean 275.49 ng) with TG and PL being the major energetic and structural lipids, respectively. Larval lipid stores were not appreciably depleted by metamorphosis and so were available for the early benthic stage juvenile. Juveniles started their benthic existence with 314 ng total lipid (TG: mean 46.84 ng, 14.9% of total, PL: mean 137.51 ng, 43.67% of total). Nile Red histochemistry and histology showed that the stomach serves as a nutrient storage organ and, that lipid stores accrued by larvae sustain developing juveniles for up to 4 days post settlement. Triglyceride supported both non-feeding stages of development and the prefeeding larval and perimetamorphic benthic stage. In this first study of lipid stores in settlement stage echinoderm larvae, we show that T. gratilla larvae sequester the same major energetic lipid (TG) to support the early juvenile that the female parent provided them to fuel early development.     

Differential utilization of biochemical components during larval development of the spider crab Maja brachydactyla (Decapoda: Majidae)

Changes in lipid class, fatty acid, fat-soluble vitamins, amino acid and minerals were studied during larval development of Maja brachydactyla Balss, 1922 in order to provide information of its ontogeny and evaluate possible dietary deficiencies and constraints. Four different batches were analyzed from hatching to metamorphosis using enriched Artemia as food. Cultured larvae were in a good nutritional condition as confirmed by the continuous lipid accumulation throughout ontogeny. A regulation of polyunsaturated fatty acids (PUFA) occurred during development in order to maintain adequate basal levels of ω-3 fatty acids (likely through a retro conversion of C₂₀ and C₂₂ into C₁₈ PUFA). Variations in the tocopherol levels indicated its possible role in PUFA protection against oxidation. Essential amino acid balance during development was not correlated with the Artemia feeding regime, but rather reflected inherent variations of the own species ontogeny. Larval requirements in essential minerals were fully supplied by the enriched Artemia.     

Effect of temperature on viability and axial muscle development in embryos and yolk sac larvae of the Northeast Arctic cod (Gadus morhua)

Cod (Gadus morhua L.) eggs may develop and hatch within temperatures of −1.5 to 12 °C, but little is known about the effects of very low temperatures on larval characteristics. Eggs of the Northeast Arctic cod (Gadus morhua) were incubated at 1, 5 or 8 °C from Day 1 after fertilisation until hatching, and transferred to 5 °C after hatching. Histological samples of the axial musculature were taken at hatching and 5 d after hatching, and the data on muscle cellularity from these samples were related to survival and hatching, size, developmental data and viability of the yolk sac larvae. All larvae hatched at the same developmental stage. Incubation of eggs at 1 °C produced shorter larvae with a larger yolk sac and more, small deep fibres at hatching than larvae from eggs incubated at 5 or 8 °C. The larval size difference was still present 5 d after hatching, a time at which the larvae from 1 °C-incubated eggs were less developed and less resistant to an acute viability stress test (65 ppt salinity). Although there were no differences between temperature groups in number and size of muscle fibres 5 d after hatching, the deep fibres of the 1 °C-group contained less myofibrils than the two other groups. The phenotype of the larvae at hatching was thus affected within these incubation temperatures. Although all groups were transferred to the same temperature after hatching, the lowest egg incubation temperature (1 °C) still had a negative effect 5 d after hatching, as these larvae were both smaller, less resistant to stress and had less functional muscles at the time of first feeding. Our conclusion is therefore that 1 °C is close to, or below, the lower thermal tolerance limit for normal functional development of Northeast Arctic cod. The results are discussed in relation to larval viability and recruitment of this species in the wild. Received: 4 February 1998 / Accepted: 10 July 1998     

Ladybird mothers mitigate offspring starvation risk by laying trophic eggs

A large proportion of ladybird beetle (Coleoptera: Coccinellidae) eggs are apparently infertile—they do not develop an embryo and are consumed by larvae hatching within the egg batch. The predicted benefits of egg consumption for larvae are empirically well supported. An important question, however, remains: are these eggs a maternal strategy to feed offspring (i.e., trophic eggs) or did egg eating evolve to exploit unavoidably infertile eggs? We investigated the adaptive value of infertile eggs in laboratory experiments with multicoloured Asian ladybirds (Harmonia axyridis). Female H. axyridis were assigned to low and high resource environments for brief intervals; we predicted that tactics to facilitate egg cannibalism, such as infertile egg production and hatching asynchrony, would be adopted in low food environments in which starvation risk for offspring is greater. We conducted two experiments in this manner that provided females with information about resource levels through prey feeding or scent. We also observed female oviposition patterns and tested for infertile egg distributions that departed from random. Females produced 56% more infertile eggs in the low vs. the high food treatment; however, hatching synchrony did not change. We consider a potential confound between information and nutrition state unlikely because ladybirds are well able to tolerate low food for 24 h, the duration of trials, and because females were in good condition when trials began. Results suggest that ladybirds use information from prey encounter to manipulate the proportion of trophic eggs in a manner consistent with the adaptive hypothesis, the first evidence of trophic egg plasticity in a non-eusocial insect.