The influence of irradiance on the biochemical composition of three phytoplankton species and their nutritional value for larvae of the Pacific Oyster (Crassostrea gigas)

Three species of phytoplankton grown at high (HL) or low light (LL) were fed as saturating rations to laboratory-reared larval Crassostrea gigas. Larval C. gigas fed diets of HL grown Chaetoceros gracilis and HL grown Isochrysis aff. galbana grew faster than those fed LL grown cells of the same phytoplankton species. Faster growth of C. gigas larvae was consistently associated with increases in the percent composition of short chain saturated fatty acids (FA) 14:0+16:0 in the HL grown cells. There were no consistent and significant differences between HL and LL grown phytoplankton cells in their content of carbon, nitrogen, protein, lipid or carbohydrate. Intraspecific increases in percent composition of essential fatty acids (EFAs), 20:53 and 22:63, in the phytoplankton were not associated with improvements in the growth or survival of the oyster larvae. Oyster larvae fed diets of Phaeodactylum tricornutum with a relatively high proportion of EFAs grew more slowly than those fed C. gracilis. In this experiment the proportion of dietary EFA 20:53 was negatively correlated with oyster growth rates. The faster growing oyster larvae contained relatively more of the FAs 14:0+16:0 which may be useful as measures of larval oyster condition. After a diet of one phytoplankton species for ca. 10 d, oyster larvae acquired distinctive FA profiles resembling that of their phytoplankton prey.     

Effects of monospecific algal diets of varying biochemical composition on the growth and survival of Pacific oyster (Crassostrea gigas) larvae

Using monospecific diets of Thalassiosira pseudonana cells grown under different steady-state conditions, it was determined that higher growth rates of larval Crassostrea gigas Thunberg were obtained when fed T. pseudonana cells grown under high light. High light grown T. pseudonana cells consistently contained relatively more of the saturated fatty acids 14:0 and 16:0. Considered over three independent experiments, high light grown T. pseudonana cells were lower in protein and higher in carbohydrate than low light grown cells. Higher growth rates of larval C. gigas were obtained on diets with more of the essential fatty acid (EFA) 22:63, and less of the other EFA, 20:53. The relative requirements of C. gigas larvae for the essential fatty acids 20:53 and 22:63 are discussed. Faster growing larvae contained higher percentages of the fatty acids 14:0 and 16:0, and lower percentages of 22:2j. Oyster growth rates were correlated with their content of the fatty acids: 14:0, 16:0 and 22:2j in two experiments utilizing separately spawned batches of larvae. Fatty acid profiles are proposed as a technique for assessing larval condition. C. gigas larvae contained ten times the percent composition of the FAs 16:43, 18:17, 20:17 and 22:2j compared with their diet. Correlation analysis suggests that the dietary source of 18:17, 20:17 and 22:2j was 16:17. It is concluded that T. pseudonana cells grown under high light are a superior diet for C. gigas larvae in comparison with low light grown cells of the same species.     

Importance of the fatty acids 20:5ω 3 and 22:6ω 3 in the diet of plaice (Pleuronectes platessa) larvae

The requirement of plaice (Pleuronectes platessa L.) larvae for the fatty acids 20:5 3 and 22:6 3 was studied. Larvae were reared from first feeding to beyond metamorphosis on Artemia sp. nauplii (EG brand), whose nutritional content had been manipulated by enrichment. Some larvae were fed Artemia rich in 20:5 3 and 22:6 3, and others were fed nauplii with no 22:6 3 and low levels of 20:5 3 (1.9% of the total fatty acids). The differences in diet had no significant difference on growth or survival of the plaice larvae. The results indicate that the plaice larvae in this study did not require 22:6 3 in their diet, even when the levels of 20:5 3 were low.     

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.     

Effects of food quality on the reproductive success of Acartia tonsa and Acartia hudsonica: laboratory observations

The effect of the chemical composition of food on the reproductive success of the copepods Acartia tonsa Dana and A. hudsonica Pinhey was studied in the laboratory. Laboratory-reared individuals were fed one of three monoalgal diets at different stages of growth: the diatom Thalassiosira weissflogii, the flagellate Rhodomonas lens and the dinoflagellte Prorocentrum minimum. The diet was analyzed for carbon, nitrogen, protein, carbohydrate and fatty acid content. Reproductive success was measured as eggs female^-1 day^-1 (E _r) and as the hatching success of the eggs. The E _r of Acartia spp. was correlated with protein and specific fatty acids [16:17 (negative), 20:53, 22:63, and 18:0 (positive)] and, especially, the fatty acid composition of the algae expressed as the 3:6 and 20:22 fatty acid ratios. The youngest diatom cultures and exponentially-growing flagellates displayed the highest E _r; the lowest E _r was recorded for females fed the senescent diatom cultures. The development time of eggs was affected by the age of the phytoplankton culture fed to the female. Hatching success of eggs decreased with the age of the algal culture, but no correlation was found with the meansured chemical components of the food.     

Incorporation and utilization of bacterial lipids in theSolemya velum symbiosis

We undertook a detailed analysis of the lipid composition ofSolemya velum (Say), a bivalve containing endosymbiotic chemoautotrophic bacteria, in order to determine the presence of lipid biomarkers of endosymbiont activity. The symbiont-free clamMya arenaria (L.) and the sulfur-oxidizing bacteriumThiomicrospira crunogena (Jannasch et al.) were analyzed for comparative purposes. The ¹³C ratios of the fatty acids and sterols were also measured to elucidate potential carbon sources for the lipids of each bivalve species. Both fatty acid and sterol composition differed markedly between the two bivalves. The lipids ofS. velum were characterized by large amounts of 18: 17 (cis-vaccenic acid), 16:0, and 16 : 17 fatty acids, and low concentrations of the highly unsaturated plant-derived fatty acids characteristic of most marine bivalves. Cholest-5-en-3-ol (cholesterol) accounted for greater than 95% of the sterols inS. velum. In contrast,M. arenaria had fatty acid and sterol compositions similar to typical marine bivalves and was characterized by large amounts of the highly unsaturated fatty acids 20 : 53 and 22 : 63 and a variety of plant-derived sterols. The fatty acids ofT. crunogena were similar to those ofS. velum and were dominated by 18:17, 16:0 and 16:17 fatty acids. Thecis-vaccenic acid found inS. velum is almost certainly symbiontderived and serves as a potential biomarker for symbiontlipid incorporation by the host. The high concentrations ofcis-vaccenic acid (up to 35% of the total fatty acid content) in both symbiont-containing and symbiont-free tissues ofS. velum demonstrate the importance of the endosymbionts in the lipid metabolism of this bivalve. The presence ofcis-vaccenic acid in all the major lipid classes ofS. velum demonstrates both incorporation and utilization of this compound. The ¹³C ratios of the fatty acids and sterols ofS. velum were significantly lighter (–38.4 to –45.3) than those ofM. arenaria (–23.8 to – 24.2) and were similar to the values found for the fatty acids ofT. crunogena (–45); this suggests that the lipids ofS. velum are either derived directly from the endosymbionts or are synthesized using endosymbiontderived carbon.Woods Hole Oceanographic Institution Contribution No. 7356Please address all correspondence and reprint requests to Dr Conway at her present address: Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260, USA     

Biochemical composition of Nyctiphanes australis and its possible use as an aquaculture feed source: Lipids,pigments and fluoride content

Nyctiphanes australis contained, on a dry weight basis, an average of 52% crude protein and 5.0 to 9.5% lipid. The fatty acid profile of N. australis was markedly unsaturated, with a mean total 3 fatty acid content of 48.6±2.4% of total fatty acids. N. australis contained high levels of the essential long-chain polyunsaturated fatty acids eicosapentaenoic (EPA, 20:53) and docosahexaencic (DHA, 22:63), ranging from 16.6 to 36.5% and 11.1 to 24.8%, respectively. The concentration of total carotenoids ranged from 137 to 302 g g^–1 dry wt, with no significant differences in concentrations found with season or life stage. The carotenoids were comprised of 79.5% astaxanthin and 20.5% canthaxanthin. The lipid and pigment compositions of N. australis suggest that the species could serve as a suitable feed source for cultured salmonids. Like other euphausiids, N. australis contained high levels of fluoride, with a seasonal range between 277 and 3507 g g^–1 dry wt. The high fluoride levels found in N. australis would not detract from its potential as a feed source for salmonids because ingested fluoride is largely absorbed by the skeleton.     

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.     

Impacts of copepods on marine seston,and resulting effects on<Emphasis Type="Italic"> Calanus finmarchicus</Emphasis> RNA:DNA ratios in mesocosm experiments

We investigated the impact of copepods on the seston community in a mesocosm set-up, and assessed how the changes in food quantity, quality and size affected the condition of the grazers, by measuring the RNA:DNA ratios in different developmental stages of Calanus finmarchicus. Manipulated copepod densities did not affect the particulate carbon concentration in the mesocosms. On the other hand, chlorophyll a content increased with higher copepod densities, and increasing densities had a positive effect on seston food quality in the mesocosms, measured as C:N ratios and 3:6 fatty acid ratios. These food quality indicators were significantly correlated to the nutritional status of C. finmarchicus. In contrast to our expectations, these results suggest a lower copepod growth potential on higher quality food. However, in concordance with earlier studies, we found that when copepods were in high densities the large particles (>1000 µm³) decreased and that the smaller particles (<1000 µm³) increased in number. These patterns were closely linked to the condition of C. finmarchicus, which were of better condition (RNA:DNA ratios) with increasing biovolumes of large particles, and, conversely, lower RNA:DNA ratios with increasing biovolumes of smaller particles. Consequentially, the selective grazing by copepods stimulated increased biovolumes of smaller plankton, and this increase was responsible for the increased food quality, in terms of C:N and 3:6 ratios. Thus, we conclude that the decreasing growth potentials of C. finmarchicus were a result of a decrease of favourably sized food particles, induced by copepod grazing.Communicated by O. Kinne, Oldendorf/Luhe     

Patterns of photosynthetic carbon metabolism in light-limited phytoplankton

Patterns of phytoplankton carbon (C) metabolism were examined in å combined laboratory and field study to assess the influence of light conditions on ¹⁴C assimilation into photosynthetic end-products. Laboratory studies with three species representing distinct size classes and taxonomic groups tested the influence of low light on patterns of C flow. Prorocentrum mariae-lebouriae (dinoflagellate) and Ditylum brightwellii (diatom) showed decreased movement of photoassimilated ¹⁴C into protein following a shift to low light ¹⁴C assimilation into lipids and photosynthetic pigments increased in low light and was paralleled by increased chl a per cell. The proportion of ¹⁴C fixed into protein returned to the pre-shift level upon return to initial light conditions. Monochrysis lutheri (chrysophyte) did not show this pattern of reduced % ¹⁴C protein. Incubations of 12 and 24 h demonstrated significant rearrangements in labeling patterns at night, wherein ¹⁴C flow into protein in darkness was favored. % ¹⁴C protein at night was lower for M. lutheri than for the other species, suggesting some interspecific differences in the low light response. Measurements of ¹⁴C assimilation in phytoplankton assemblages from Chesapeake Bay demonstrated movement of a higher proportion of photo-assimilated C into protein in samples collected in the surface mixed layer than in those below the pycnocline. In comparison, phytoplankton collected below the pycnocline fixed a higher proportion of ¹⁴C into lipids, photosynthetic pigments, and low molecular weight metabolites, as was observed in low light laboratory cultures. A comparison of 12- and 24-h incubations for measuring patterns of C flow into photosynthetic end-products confirmed the inadequacy of short-term measurements, as significant changes in ¹⁴C allocation occurred in the dark phase of the photocycle. Together, these results suggest that ¹⁴C assimilation into photosynthetic end-products can be a useful measure of adaptive state in changing light conditions, but point out some difficulties in applying this approach in situ.     

Metabolism of palmitic,linoleic, and linolenic acids in adult oysters,Crassostrea virginica

This study investigated incorporation and metabolism of saturated [(1-¹⁴C) 16:0] and unsaturated [(1-¹⁴C) 18:26 and (1-¹⁴C) 18:33] fatty acids in adult eastern oysters,Crassostrea virginica Gmelin (spawned from parents obtained in 1986 from Mobjack Bay, Virginia, USA), and the influence of temperature on these processes. InC. virginica, incorporation of injected palmitic (16:0) and linolenic (18:33) acids was increased when oysters which had been grown in warm water (22 to 23°C) were transfered to cold water (5 to 7°C) for 8 to 18 d. Incorporation of linoleic acid (18:26) was unchanged under these conditions. The changes in concentration may have been linked to depression of metabolism in these oysters, in particular that of 16:0, which was reduced by 90%. Oxidation of incorporated fatty acids was much higher in warm than in cold water. Cold-temperature conditioning ofC. virginica altered the distribution of fatty acids among the neutral and polar lipid fractions. Long-term exposure to cold water increased the proportion of fatty acids in the polar fraction, which may be related to maintenance of membrane fluidity. Short-term exposure to cold water had the opposite effect, which may be due to increased energy requirements as the oyster adapts to new conditions. Reutilization of¹⁴C-acyl groups demonstrated de novo synthesis of 16:0 and 18:0 fatty acids. Only limited elongation and no desaturation of the administered fatty acids was observed.     

Modeling Free-Surface Fluctuation Effects for Calculation of Turbulent Open-Channel Flows

A new method of introducing the free-surface effects in the calculation of turbulent open-channel flows using the amplitude of the free-surface fluctuation is proposed along with a modeling method of the equation for the free-surface fluctuation. It can be incorporated in two-equation models like k-or k-type models by introducing the damping factor to represent the interaction of the eddies with the fluctuating free-surface. Test calculations for fully developed flows and those over backward-facing step indicate good agreement with direct numerical simulation results as well as experimental results.     

Expansive covers of turf-forming algae on human-dominated coast: the relative effects of increasing nutrient and sediment loads

Turf-forming algae form more extensive habitat on subtidal rock adjacent to urban than non-urban coast of South Australia. This pattern is frequently observed on the worlds temperate coasts and is variously considered to be a result of enhanced concentration of nutrients or rates of sediment deposition on urban coasts. We experimentally tested which of three components of environmental change (increased nutrients in water, increased nutrients in sediments and increased sediment deposition) best explain the expansive covers of turf-forming algae on urban coasts. All three treatments had independent and positive effects on the percentage cover of turf-forming algae. The addition of nutrients from the water column had the largest influence (²=0.55), which was more than six times greater than the effect of nutrients added to sediments (²=0.08). An increase in rate of deposition of sediments had substantial effects (²=0.35), which were about one third less than those of water-borne nutrients. Importantly, the combined effect of all three treatments caused a 77% increase in percentage cover of turf-forming algae, which is comparable to the observed difference in covers between urban and non-urban coast in South Australia (93%). These results suggest that human activities that reduce water quality in both nutrient and sediment loads account for major change observed on human-dominated coasts. Despite this knowledge, we still lack complete information on the mechanisms that switch the primary subtidal habitat from canopy-forming algae to turf-forming algae on human-dominated coasts.Communicated by M.S. Johnson, Crawley     

Changes in the digestive gland of Euphausia superba during short-term starvation: lipid class,fatty acid and sterol content and composition

During a period of short-term (19 d) starvation, total lipid in the digestive gland of Euphausia superba Dana decreased from 21 to 9% dry weight. Total lipid per digestive gland decreased significantly during starvation compared to Day 0 individuals, falling from 1960 (±172) to 385 (±81) g. Polar lipid was the major lipid class utilised during starvation, falling from 1510 (±225) to 177 (±46) g per digestive gland (76 to 45%). Absolute levels of triacylglycerol fell from 300 (±41) to 76 (±5) g; however, relative levels remained unchanged. The relative level of free fatty acid increased significantly with starvation (4 to 39%) with absolute levels ranging from 79 (±1) to 156 (±20) g per digestive gland. Absolute levels of all fatty acids per digestive gland declined continually until the end of the starvation period. The long-chain polyunsaturated acids eicosapentaenoic (20:53) and docosahexaenoic (22:63), decreased with starvation from 37 to 26% and 15 to 10%, respectively whereas the saturated fatty acid, palmitic acid (16:0), increased from 15 to 20%. Cholesterol, the major sterol in this organ, increased from 17 (±20) to 44 (±13) g per digestive gland by Day 3, and by Day 19 had returned to levels found in the digestive gland of Day 0 individuals. Desmosterol followed a similar pattern to cholesterol, increasing from 3 (±1) g per digestive gland on Day 0 to 11 (±4) g on Day 3, and falling to 2 (±1) g on Day 19. Other sterols in the digestive gland, predominantly of algal origin, fell from the levels found in Day 0 individuals to near zero amounts by Day 6. The digestive gland of E. superba plays a dynamic role during shortterm starvation in terms of lipid content and composition. The relative levels of polar lipids, free fatty acids and cholesterol in the digestive gland may provide reliable indices of the nutritional condition of E. superba in the field. Sterols in the digestive gland are indicative of recent dietary composition of krill, and may also be used to quantify dietary input from individual phytoplanktonic species.     

Marine lipids in a cold coastal ecosystem: Gilbert Bay,Labrador

Some organisms living in coastal bays in Newfoundland and Labrador have to contend with sub-zero temperatures for most of the year. The goal of this study was to examine the lipid composition of the food web in such an environment in order to obtain information on essential nutrients and trophic relations. In August 2000, plankton, 16 species of macroinvertebrates, and sediments were collected in a shallow, sheltered bay, Gilbert Bay, southern Labrador (52°35N; 55°50W). Plankton had higher proportions of 3 fatty acids (39±5%) and lower proportions of bacterial fatty acids (5±2%), while sediments had 12±5% 3 fatty acids and 15±9% bacterial fatty acids. Plankton 3 fatty acids levels and sediment bacterial fatty acid levels were higher than in equivalent samples previously collected further south, in Newfoundland. Benthic macroinvertebrates contained 0.5±0.4% of their wet weight as lipid, of which 45±15% was triacylglycerol. Levels of 3 fatty acids were high and relatively constant across phyla, accounting for 36±11% of the total fatty acids. Compared to other invertebrates, echinoderms had a unique fatty acid composition, which was characterized by high levels of 20:4-6 (9±6%), bacterial fatty acids (8±4%), and 20:1+22:1 monoenoic fatty acids (13±6%). Baseline lipid data from plankton and macroinvertebrates showed constantly high levels of polyunsaturated fatty acids, indicating their physiological importance at low temperatures.Communicated by J.P. Grassle, New Brunswick     

Feeding deterrence properties of apo-fucoxanthinoids from marine diatoms. I. Chemical structures of apo-fucoxanthinoids produced by Phaeodactylum tricornutum

Methanol extracts of freshly harvested cells of the marine diatom Phaeodactylum tricornutum were found to produce a feeding deterrent effect in the copepod Tigriopus californicus. Bioassay guided fractionation of the methanol extracts led to the isolation of four compounds possessing feeding deterrent activity. The compounds were identified as apo-10-fucoxanthinal (1), apo-12-fucoxanthinal (2), apo-12-fucoxanthinal (3), and apo-13-fucoxanthinone (4) by detailed spectroscopic analysis and comparison with authentic compounds produced semi-synthetically from fucoxanthin. Compounds 1 to 4 exhibited feeding deterrent responses in T. californicus at concentrations of less than 20 ppm.     

Swimming pattern as an indicator of the roles of copepod sensory systems in the recognition of food

The roles of copepod sensory systems in the recognition of food were investigated using the Bugwatcher, a video-computer system designed to track and describe quantitatively the swimming patterns of aquatic organisms. The swimming behavior of the copepodPseudocalanus minutus in the presence of phytoplankton is characterized by a decrease in average swimming speed and an increase in pause behaviors compared to its swimming behavior in filtered seawater. Copepods exposed to chemosensory stimulation alone (filtered phytoplankton exudate) exhibited an increase in average swimming speed and an increase in the number of burst swimming behaviors. When exposed to a novel, non-food chemosensory stimulus (morpholine), no change in swimming behavior was observed unless the copepods had been conditioned to this odor in the presence of phytoplankton. Copepods exposed to mechanosensory stimulation alone (plastic spheres) exhibited a decrease in swimming speed and an increase in pause behaviors. When exposed to both forms of stimulation simultaneously (phytoplankton exudate and plastic spheres), a further decrease in swimming speed and increase in pause behaviors occurs, yielding a swimming pattern similar to that found in the presence of phytoplankton. This analysis of swimming pattern indicates that both chemoreception and mechanoreception contribute to the recognition of food inP. minutus.Contribution No. 406 of the US EPA Environmental Research Laboratory; Narragansett, Rhode Island 02882, USA     

Primary production in Dumbell Bay in the Arctic Ocean

Photosynthesis, chlorophyll, inorganic nutrients, and related factors were measured throughout a productive season in a small coastal inlet of the Arctic Ocean. Significant production was confined to 5 m for a very limited time period beacause of both light and nutrient limitations. Maximum chlorophyll was 8.2 mg Chl a m^-3 and maximum gross and net photosynthesis rates were 830 and 550 mg C m^-2 d^-1. Annual gross and net photosynthesis is estimated at about 12 and 9 g C m^-2 respectively. The effect of light stimulation on assimilation is described, sources of coastal zone nutrient enrichment are considered, and discrepancies between gross and net photosynthesis are discussed with reference to the growth characteristics of the phytoplankton population.Bigelow Laboratory for Ocean Sciences. Contribution No. 76008     

Variation in lipid classes during the molting cycle of the prawn Penaeus japonicus

The variation in the concentration and fatty acid composition of lipid classes during the molting cycle of the prawn Penaeus japonicus was investigated. The lipid concentration of the whole body reached a maximum at mid-premolt (Stage D₂) and then decreased to low level at late premolt (Stage D_3–4). The accumulation of lipids during the premolt period seemed to be attributable to the increase of both polar and neutral lipids. The increase of neutral lipids at Stage D₂ was derived from not only triglycerides but also free sterols and free fatty acids. Regarding the fatty acid composition of every lipid class, a marked variation occurred mainly at the intermolt (Stage C). In this stage, the polar lipids were rich in monoenoic acids such as 18:1 and poor in polyenoic acids such as 20:53 and 22:63. The triglycerides were rich in polyenoic acids at Stage C, but poor in monoenoic acids such as 16:1 and 18:1. The steryl esters contained large amounts of saturated acids such as 16:0 and 18:0 throughout the molting cycle, however the level of polyenoic acids increased at Stage C.     

Physiological ecology of picoplankton in the North Sea

The distribution of cyanobacteria in the surface waters of the North Sea was measured during July 1987. Numbers of cyanobacteria ranged from 2.5x10⁶ to 1.7x10⁸ cells 1^-1. In the majority of stations, cyanobacterial numbers were highest in the near-surface water and a subsurface maximum was found at only one station. The distribution of ¹⁴C among the end-products of photosynthesis was determined for picoplankton (<1 m) and other phytoplankton >1 m throughout the North Sea. The majority of label was found in the protein fraction of both picoplankton and >1 m phytoplankton; incorporation into lipids and polysaccharides plus nucleic acids was much lower. We interpret the large incorporation into protein to be a consequence of nutrient limitation of these natural assemblages. Photosynthetic parameters of the two size fractions were also determined. Assimilation number (P _m ^B ) and initial slope were greater for the picoplankton fraction than for phytoplankton >1 m but there was no evidence of significant photoinhibition of either fraction at irradiances up to 1 000 E m^-2 s^-1.