Lipid classes and their content of n-3 highly unsaturated fatty acids (HUFA) in Artemia franciscana after hatching, HUFA-enrichment and subsequent starvation

The distribution of n-3 highly unsaturated fatty acids (HUFA) over the major neutral and polar lipid classes was determined for two predominant types of live food used in the larviculture of marine fish and shrimp, i.e. freshly hatched and HUFA-enriched Artemia, and compared with data reported in the literature for wild copepods, representing the natural diet of these larvae. Lipid class composition and their content of n-3 HUFA, particularly eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), were assessed in freshly hatched, HUFA-enriched and subsequently starved Artemia franciscana. The n-3 HUFA enrichment was based on feeding Artemia a lipid emulsion in which either fatty acid ethyl esters (EE, diluted with olive oil) or triacylglycerol (TAG) provided a level of 30% n-3 HUFA. Enrichment of Artemia with either type of the lipid emulsions resulted in an increase of total lipid content from 20.0 to 28.2–28.7% of dry matter mainly due to the accumulation of neutral lipid, primarily TAG (from 82 to 158 mg g⁻¹ dry wt in freshly hatched and 24-h enriched Artemia). Enriched brine shrimp utilized up to 27–30% of their TAG content during 72 h of starvation at 12 °C. The absolute tissue concentrations of polar lipids remained constant at 71 to 79 mg g⁻¹ dry wt throughout the enrichment and subsequent starvation. The level of n-3 HUFA increased drastically during enrichment from 6.3% of total fatty acids (8.2 mg g⁻¹ dry wt) in freshly hatched nauplii to between 20.4 and 21.8% (40.4 to 43.2 mg g⁻¹ dry wt) in 24-h enriched Artemia and was not significantly affected by the source of n-3 HUFA. During starvation, 18:0, 20:4n-6 and 20:5n-3 were retained, whereas 18:4n-3, 22:5n-3 and 22:6n-3 were specifically catabolized. The major polar lipids, phosphatidylethanolamine (PE) and phosphatidylcholine (PC), of freshly hatched Artemia showed very low levels of DHA (<0.1% of total fatty acids) and carried about 45% of the total EPA present. Enrichment with either of the emulsions resulted in an increase of the neutral lipid fraction which concentrated >64% of the EPA and >91% of the total DHA present. This is in sharp contrast with the high levels of n-3 HUFA, in particular DHA, in the polar lipid fraction reported for wild copepods. The contrasting distribution of DHA in the neutral and polar lipid fractions of enriched brine shrimp compared to the natural diet may influence the efficacy of this essential fatty acid for marine fish larvae in aquaculture systems. Received: 10 June 1997 / Accepted: 8 August 1997     

Effect of sexual maturation on the tissue biochemical composition of<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> and<Emphasis Type="Italic"> O</Emphasis>.<Emphasis Type="Italic"> defilippi</Emphasis> (Mollusca: Cephalopoda)

Changes in the protein, lipid, glycogen, cholesterol and energy contents, total amino acid and fatty acid profiles of Octopus vulgaris and O. defilippi tissues (gonad, digestive gland and muscle) during sexual maturation (spermatogenesis and oogenesis) were investigated. Both species showed an increase of amino acids and protein content in the gonad throughout sexual maturation (namely in oogenesis), but allocation of these nitrogen compounds from the digestive gland and muscle was not evident. The major essential amino acids in the three tissues were leucine, lysine and arginine. The major non-essential amino acids were glutamic acid, aspartic acid and alanine. With respect to carbon compounds, a significant increasing trend (P<0.05) in the lipid and fatty acid contents in the three tissues was observed, and, consequently, there was also little evidence of accumulated lipid storage reserves being used for egg production. It seems that for egg production both Octopus species use energy directly from food, rather than from stored products. This direct acquisition model contrasts with the previous model for Octopus vulgaris proposed by ODor and Wells (1978: J Exp Biol 77:15–31). Most of saturated fatty acid content of the three tissues was presented as 16:0 and 18:0, monounsaturated fatty acid content as 18:1 and 20:1 and polyunsaturated fatty acid content as arachidonic acid (20:4n-6), eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3). Though cholesterol is an important precursor of steroid hormones, this sterol content exhibited variations that do not seem to be related with the maturation process. Moreover, significant differences (P<0.05) were obtained between genders, suggesting that perhaps there is a greater physiological demand for cholesterol during spermatogenesis than oogenesis. If the component sterols of octopus are of a dietary origin, considerable variation in the cholesterol content between species might be expected on the basis of the sterol composition of their prey. The glycogen reserves increased significantly in the gonad and decreased significantly (P<0.05) in the digestive gland and muscle of O. vulgaris (these trends were not evident in O. defilippi). Glycogen may play an important role in the maturation process and embryogenesis of these organisms, because carbohydrates are precursors of metabolic intermediates in the production of energy. It was evident that sexual maturation had a significant effect upon the gonad energy content, but the non-significant energy variation (P>0.05) in the digestive gland and muscle revealed no evidence that storage reserves are transferred from tissue to tissue. The biochemical composition of digestive gland and muscle may not be influenced by sexual maturation, but rather by other biotic factors, such as feeding activity, food availability, spawning and brooding.Communicated by S.A. Poulet, Roscoff     

Changes in phospholipase A2 activity and lipid content during early development of Atlantic halibut (Hippoglossus hippoglossus)

During early development in fish, phospholipase A₂ (EC 3.1.1.4) regulates membrane lipid modifications, which relates to changes in environmental conditions and provision of fatty acids required for metabolic energy substrates and prostaglandin biosynthesis. A method to analyze phospholipase A₂ in rat tissues has been modified to measure its activity in embryonic Atlantic halibut (Hippoglossus hippoglossus L.). Egg and embryo samples were collected during the 1994 spawning season. Enzyme activity was undetectable at fertilization but in 10-d embryos was 230 pmol mg⁻¹ h⁻¹ (at 20 °C) and increased by ∼120% at hatch (17-d). Significant alterations in the fatty acid composition of important phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), were also observed. The content of some critical polyunsaturated fatty acids, and the ratio of unsaturated/saturated fatty acids, declined significantly over development. Acyl-chain restructuring mediated through the activity of phospholipase A₂, coupled with other observed lipid changes (significant increases in the PC/PE ratio and cholesterol content), would produce a decreased fluidity of membranes during embryonic development, coinciding with the predicted upward movement of larvae in the water column. Arachidonic acid (20:4n-6) removed from PE could serve as a precursor for biosynthesis of 2-series prostaglandins, and eicosapentaenoic acid (20:5n-3) from PC is a likely source for other prostaglandin types. Despite removal of polyunsaturated fatty acids, there was an overall increase in lipid and fatty acid concentration, which can be attributed to amino acid catabolism during early developmental stages. Received: 9 September 1996 / Accepted: 8 September 1997     

Ecological investigations on the zooplankton community in balsfjorden,northern Norway: Lipids and fatty acids in Meganyctiphanes norvegica,Thysanoessa raschi and T. inermis during mid-winter

Total lipid of Meganyctiphanes norvegica (M. Sars) contained 53% triacylglycerols and traces of wax esters, that of Thysanoessa raschi (M. Sars) contained 44% triacylglycerols and 10% wax esters and that of T. inermis (Krøyer) contained 28% triacylglycerols and 40% wax esters. The triacylglycerols of M. norvegica were relatively rich in 20:1 and 22:1 fatty acids and its traces of wax esters resembled those of calanoid copepods. The triacylglycerols of both Thysanoessa species were deficient in 20:1 and 22:1 fatty acids but were richer in 16:1(n-7) and 18:1 (n-7) acids than those of M. norvegica. The wax esters of T. raschi contained phytol as almost the only fatty alcohol and were rich in 16:0 and 18:1 (n-9) fatty acids. The wax esters of T. inermis contained mainly 16:0 and 14:0 fatty alcohols with lesser amounts of phytol and their dominant fatty acid was 18:1, especially the (n-9) isomer. The triacylglycerols of T. inermis had 18:4 (n-3) as the major polyunsaturated fatty acid. From these and other aspects of fatty acid and fatty alcohol analyses it is concluded that a major foodstuff of M. norvegica in Balsfjorden is wax ester-rich calanoid copepods. T. raschi and especially T. inermis are concluded to have much more preference for phytoplanktonic food. Results are discussed in terms of current knowledge of the lipid chemistry of krill in the northern and southern hemispheres.     

Biochemical composition of cephalopods with different life strategies,with special reference to a giant squid,<Emphasis Type="Italic"> Architeuthis</Emphasis> sp.

The present study is a comprehensive comparison of the biochemical composition (protein, lipid, glycogen and cholesterol contents, and amino acid and fatty acid profiles) of 14 species of cephalopods with different life strategies (benthic, nektobenthic, benthopelagic and pelagic) in distinct habitats (neritic, demersal, oceanic and deep sea), with special emphasis placed on a male Architeuthis sp. The giant squid showed a significantly lower protein and total amino acid content in the gonad and digestive gland. The major essential amino acids were leucine, lysine and arginine. The major non-essential amino acids were glutamic acid, aspartic acid and proline. The benthopelagic cirrate octopus Opisthoteuthis sp. showed a lower content of these nitrogen compounds in the muscle. In respect to lipid and fatty acid contents, the giant squid showed the highest values in the gonad and digestive gland and the lowest in the muscle. Most of saturated fatty acid content was presented as 16:0; monounsaturated fatty acid content, as 18:1 and 20:1; and polyunsaturated fatty acid content, as 20:4n-6, 20:5n-3 and 22:6n-3. The highest cholesterol content in the gonad was detected in Opisthoteuthis sp. and Architeuthis sp.; in the digestive gland. in Todarodes sagittatus; and in the muscle, in Sepia elegans. The highest glycogen value in the gonad was detected in Octopus vulgaris; in the digestive gland and muscle, the highest values were attained in Opisthoteuthis sp. The relationships between life strategies and biochemical composition were investigated and principal component analysis (PCA) was performed. The different cephalopod life strategies could be well separated on the basis of the first two principal components. The nektobenthic and pelagic strategies were clearly separated from the benthic, suggesting that these groups of species are characterized by lower lipid content and higher protein content in the gonad. A rather similar life-strategy distinction was obtained for the digestive gland. The benthopelagic strategy was also well separated from benthic and pelagic strategies and from Architeuthis sp. In the muscle, the results indicated lower nitrogen and carbon compound contents in Architeuthis sp. and Opisthoteuthis sp. The environmental conditions that possibly explain the Architeuthis sp. stranding, the relationships between biochemical compositions and the life strategies of the different cephalopod species studied are discussed.Communicated by S.A. Poulet, Roscoff     

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.     

Factors determining the hatching success of Antarctic krill <Emphasis Type="Italic">Euphausia superba</Emphasis> embryo: lipid and fatty acid composition

The present study addresses the effect of maternal diet on hatching success and condition of embryos and larvae of Antarctic krill Euphausia superba. Lipid and fatty acid content and composition were determined in field and laboratory samples. Developmental stages analyzed in embryos included: multiple-cell, gastrula, and limb-bud stages. Larval stages analyzed included: nauplius I, nauplius II, and metanauplius. Laboratory-reared embryos were spawned by gravid females incubated under three feeding groups: (1) phytoplankton mixture, (2) phytoplankton mixture and minced clam, and (3) phytoplankton mixture, minced clam, and commercial larval food. Hatching success was highest in group 3 (100%), lowest in group 1 (0%), and highly variable in field samples (0–48%). Lipid decreased slightly in embryos during embryonic development, while large decreases in lipid were found during nauplius development. High levels of 18:2(n-6), 20:4(n-6), and 22:6(n-3) observed with group 3 samples coincided with high hatching success in krill embryos. The ratio of 22:6(n-3)/20:5(n-3) also correlated to hatching success of embryos. The fatty acid profile of embryos in group 3 was similar to that of the field-collected embryos, reflecting the contribution of the commercial larval food in the maternal diet. In our study, the maternal diet was found to influence the fatty acid composition of embryos and in turn affects the hatching success of krill. Specific polyunsaturated fatty acids appeared to play important roles in embryogenesis in krill.     

Species-specific differences in lipid composition and omnivory indices in Arctic copepods collected in deep water during autumn (North Water Polynya)

To discern species-specific patterns in omnivory indices in copepods from the North Water Polynya, lipid classes and fatty acids were determined in Calanus hyperboreus Krøyer copepodite stage 5 (CV), C. glacialis Jaschnov (CV) and Metridia longa Lubbock (females) sampled below the mixed layer during autumn 1999. Generally, M. longa contained higher proportions of triacylglycerols, polar lipids and 18:1(n-9) than the other species. M. longa also had lower relative amounts of wax esters (WE), polyunsaturated fatty acids (PUFA), 20:1(n-9), 22:1(n-11), and lower absolute (g lipid copepod^-1) and relative (% lipid) lipid levels. Unsaturation coefficients (UC; ratio of polyunsaturated to total WE) were usually lowest in M. longa. These differences probably relate to a lesser dependence on phytoplankton in M. longa, and hence more ingestion of PUFA-poor prey. Moreover, levels of the carnivory index 18:1(n-9)/18:1(n-7) were highest in M. longa. Our data support the widespread contention that M. longa is more omnivorous than C. hyperboreus and C. glacialis. Proportions of bacterial fatty acids [odd-numbered and/or branched (OBFA); 18:1(n-7)] and PUFA/SFA (SFA: saturated fatty acids) ratios were largely unrelated to feeding strategy. Relationships between relative and absolute amounts of 16:1(n-7) and 18:1(n-7) in copepods suggest that the latter fatty acid is formed in vivo by chain elongation of the former. However, elevated levels of 18:1(n-7) and OBFA in M. longa at stations dominated by the microbial loop imply that these indices can be used to track ingestion of PUFA-poor protozoans when diatoms are scarce.Communicated by J.P. Grassle, New Brunswick     

The Arctic pteropod<Emphasis Type="Italic"> Clione limacina</Emphasis>: seasonal lipid dynamics and life-strategy

During various seasons from May 2001 until July/August 2003, the lipid dynamics of the pteropod Clione limacina from Kongsfjorden, Svalbard, were investigated with respect to ontogenetic development and life-cycle. Polytrochous larvae, which were dominant in spring (April, May), composed the lipid-richest specimens of the population, with total lipid of about 50% of dry mass (%DM). Major lipid classes were triacylglycerols (TAG) and 1-O-alkyldiacylglycerol ethers (DAGE), accounting on average for 53.1 and 21.9% of total lipid, respectively. Until summer, larvae grew to adults by utilising their storage lipids. In July/August, lipids were depleted to about 10%DM due to maturation and reproduction. Almost all animals in autumn (September) were mature and able to replenish their lipid deposits by accumulating DAGE (26.7%) and TAG (39.6%). This is probably the prerequisite for successful overwintering.Principal component analysis (PCA), based on the fatty acid compositions, revealed ontogenetic differences between polytrochous larvae, and small and full-grown adults. Higher proportions of 18:4(n-3) and 14:0 were found in polytrochous larvae and smaller adults during spring. Both fatty acids were highly significantly correlated with the proportions of TAG, which were used for growth and development because they are presumably easier to metabolise. PCA also divided C. limacina specimens into DAGE-rich and DAGE-poor. We suggest that DAGE are a long-term energy store and hypothesise that they are necessary during periods of food scarcity, but may also serve as an energy source for reproduction. The fatty acids 17:1(n-8), 15:0, 16:1(n-7) and 18:1(n-7) were significantly correlated with the proportion of DAGE but not with TAG. These fatty acids, which do not originate from their only prey, Limacina helicina, are synthesised de novo. Their abundance reflects an efficient lipid production by C. limacina. Based on the results of lipid biosynthesis and accumulation in combination with the population structure, we suggest that C. limacina has at least a 2-year life-cycle in Svalbard waters.Communicated by M. Kühl, Helsingør     

Evaluation of fatty acids as biomarkers for a natural plankton community. A field study of a spring bloom and a post-bloom period off West Greenland

An investigation of the fatty acid composition of a natural arctic plankton community was carried out over two fishing banks located between 63°N and 65°N off the West Greenland coast. Samples for fatty acid analyses, species determination and biomass assessments of the plankton community were taken at the depth of fluorescence maximum. High biomass and diatom dominance during the spring bloom and low biomass and flagellate dominance in the post-bloom period were reflected by the fatty acid profiles. The total amount of fatty acid ranged from 55 to 132 µg l^-1 during the spring bloom and from 1 to 5 µg l^-1 during the post bloom. Analysis of the fatty acids showed that when the plankton was dominated by diatoms of the genera Thalassiosira and Chaetoceros, the proportions of C16:1(n-7) and C20:5(n-3) were correspondingly high. C18s, and particularly C18:1(n-9), were more abundant when the plankton was dominated by small autotrophic flagellates, primarily haptophytes. We found a good positive correlation between the common diatom marker, C16:1(n-7)/C16:0, and the biomass percentage of diatoms (r=0.742, P<0.001), as well as between the biomass percentage of flagellates and total C18 fatty acids (r=0.739, P<0.001). This supports the use of these specific fatty acids and fatty acid ratios as general biomarkers of the plankton community. However, the fatty acids are not specific enough to sufficiently characterise the composition of the plankton community, and microscopical support is needed to verify observed trends.     

Trophic ecology of Rimicaris exoculata : a combined lipid abundance/stable isotope approach

The alvinocaridid shrimp Rimicaris exoculata is an abundant component of the biota of Mid-Atlantic Ridge hydrothermal vents. To determine the nutritional strategy of this organism, we analysed the molecular abundance and carbon isotopic composition of its phospholipid fatty acids. High abundances of n-7 fatty acids (>40% total fatty acids) were observed in R. exoculata muscle tissues, in bacterial epibionts scraped from its gill bailers, and from the bacterially infested metal sulphides that the shrimp ingest. The phospholipid fatty acid abundance data indicates that the bacteria in the sulphides are closely related to the bacterial epibiota inhabiting the shrimp gill bailers, carapace and other body parts. Compound specific δ¹³C analyses of the phospholipid fatty acids gave average values of −12‰ for the epibiont bacteria and −21‰ for the sulphide bacteria. This difference may be largely due to the expression of different forms of RuBisCO (Forms I and II) which fractionate against ¹³C to different extents. Carbon limitation within the shrimp epibiont population may be an additional factor. The δ¹³C values (mean = −13‰) of the saturated and monounsaturated fatty acids isolated from the muscle tissues of R. exoculata were very close to those of the epibionts, indicating that the predominant source of dietary carbon for the shrimp is their epibionts, with a lesser contribution from free-living bacteria. The δ¹³C values (−26‰) of shrimp cholesterol were much more negative than those of the fatty acids, and this cholesterol is likely to have derived from the oceanic photic zone. Received: 26 June 1997 / Accepted: 6 November 1998     

Lipids and fatty acids of the benthic marine harpacticoid copepod<Emphasis Type="Italic"> Heteropsyllus nunni</Emphasis> Coull during diapause: a contrast to pelagic copepods

Many free-living copepods produce and store lipids prior to entering diapause (long-term dormancy). Heteropsyllus nunni Coull is the only marine harpacticoid copepod known to undergo any form of diapause. This study presents the first information on the types of lipids and fatty acids produced for long-term diapause in this benthic species. Sexually immature adults of H. nunni undergo diapause within a pliable self-made cyst. Prior to entering diapause (which lasts 3–4 months), they produce and store large amounts of orange lipid. The lipids apparently are utilized during diapause. Although some residual lipids remain, chiefly around the gonads, after the copepods emerge from their cysts, the lipid stores are visibly reduced. Typically, the copepods mate and produce eggs within 48 h after diapause is terminated. Light level and confocal laser scanning microscopy revealed that the lipid stores are distributed throughout the body in numerous oil droplets and not as a single oil sac, as seen in many marine calanoid copepods prior to overwintering (winter diapause). Transmission electron microscopy showed lipid spheres within the gut epithelium and large droplets of lipids stored extracellularly. Confocal laser scanning microscopy of copepods in pre-diapause, during diapause (encysted), post-diapause (recently excysted), and in reproductive condition, revealed that lipid stores are reduced following diapause, but are not totally absent. Analysis of lipid classes showed that H. nunni store predominantly wax esters/sterol esters (83% of total lipids) during diapause. The predominant lipid is most likely wax esters, as sterol esters typically are found only in small amounts in copepods. Fatty acid (FA) profiles of the copepods in diapause showed 16:0 to be most abundant followed by 16:1n-7 and 18:0; other FA occurred at concentrations <10% of total FA. Three polyunsaturated fatty acids (PUFA), 20:5n-3, 18:2n-6 and 20:4n-6, were found at concentrations <2% of total FA. These PUFA are "essential fatty acids" in H. nunni, obtained through dietary sources. The lipid classes and fatty acids present in H. nunni during diapause are compared to those of other copepods, some in a state of diapause and others not. It appears that lipid class and FA profiles are indicative of genetic makeup, type of diet or amount of food consumed prior to dormancy. Some classic paradigms of lipids and their association with copepod diapause are re-evaluated.Communicated by P.W. Sammarco, Chauvin     

Seasonal lipid dynamics of the copepods <Emphasis Type="Italic">Pseudocalanus minutus</Emphasis> (Calanoida) and <Emphasis Type="Italic">Oithona similis</Emphasis> (Cyclopoida) in the Arctic Kongsfjorden (Svalbard)

Seasonal lipid dynamics of various developmental stages were investigated in Pseudocalanus minutus and Oithona similis. For P. minutus, the dominance of 16:1(n−7), 16:4(n−3) and 20:5(n−3) fatty acids indicated a diatom-based nutrition in spring, whereas 22:6(n−3), 16:0, 18:2(n−6) and 18:1(n−9) pointed to a flagellate-based diet during the rest of the year as well as omnivorous/carnivorous low-level feeding during winter. The shorter-chain fatty alcohols 14:0 and 16:0 prevailed, also reflecting biosynthetic processes typical of omnivores or carnivores. Altogether, the lipid signatures characterized P. minutus as an opportunistic feeder. In contrast, O. similis had consistently high amounts of the 18:1(n−9) fatty acid in all stages and during all seasons pointing to a generally omnivorous/carnivorous/detritivorous diet. Furthermore, the fatty alcohol 20:1(n−9) reached high percentages especially in adult females and males, and feeding on Calanus faecal pellets is suggested. Fatty alcohols, as wax ester moieties, revealed significant seasonal variations in O. similis and a seasonal trend towards wax ester accumulation in autumn in P. minutus. P. minutus utilized its lipid deposits for development in the copepodite stages III and IV and for gonad maturation in CV and females during the dark season. However, CVs and females depended on the spring phytoplankton bloom for final maturation processes and reproduction. O. similis fueled gonad maturation and egg production for reproduction in June by wax esters, whereas reproduction in August/September co-occurred with the accumulation of new depot lipids. Both species revealed significantly higher wax ester levels in deeper (>50 m) as compared to surface (0–50 m) dwelling individuals related to a descent prior to overwintering.     

Seasonal patterns in fatty acids of <Emphasis Type="Italic">Calanus hyperboreus</Emphasis> (Copepoda,Calanoida) from Cumberland Sound,Baffin Island,Nunavut

The marine copepod Calanus hyperboreus accumulates large quantities of lipids and essential fatty acids during summer months in Northern oceans. However, few data exist regarding their winter fatty acid profiles, which could be informative regarding the use of lipids by C. hyperboreus to successfully survive and reproduce during times of ice-cover and limited food. The present study compared fatty acids of C. hyperboreus between summer (August 2007 and 2008) and winter (early April 2008 and 2009) in Cumberland Sound, Canada. Summer samples from both years had significantly higher ∑polyunsaturated fatty acids and unsaturation indices (based on μg fatty acid mg dry tissue⁻¹) than winter samples and separated on a principal component analysis due to higher 18:2n-6, 18:4n-3, and 20:5n-3, consistent with phytoplankton consumption. Winter C. hyperboreus had significantly higher ∑monounsaturated fatty acids (MUFA) versus summer samples and separated on the principal component analysis due to higher proportions of 16:1n-7, 20:1n-9, and 22:1n-9, suggesting they were not actively feeding. Based on the seasonal fatty acid comparison, C. hyperboreus was catabolizing specific fatty acids (e.g. 20:5n-3), conserving others (e.g. 22:6n-3), and maintaining or increasing biosynthesis of certain MUFA (e.g. 18:1n-9) during winter. These findings provide insight into the seasonal strategy of acquisition (summer) and utilization (winter) of specific fatty acids by a key Arctic organism and could become important for monitoring changes in fatty acids associated with decreased ice-cover duration due to climate warming.     

Fatty acid compositions of neutral glycerides from the ovaries of the asteroids Ctenodiscus crispatus,Asterias lincki and Pteraster militaris from Balsfjorden,northern Norway

Fatty acid analyses of lipids in the asteroids Ctenodiscus crispatus (Retzius), Asterias lincki (Müller and Troschel) and Pteraster militaris (O. F. Müller) collected in Balsfjorden, near Tromsø, northern Norway in 1978–1980, were performed to further our understanding of the asteroids' reproductive biology. Fatty acid compositions were determined in phospholipids, triacylglycerols and alkyldiacylglycerols isolated from ovaries of frozen C. crispatus. The major polyunsaturates in phospholipids were 20:4(n-6) followed by 20:5(n-3). Both the triacylglycerols and alkyldiacylglycerols were relatively deficient in polyunsaturates but had 20:1(n-13) and 18:1(n-7) as major components. Both these liqid classes had additionally 15% of their fatty acids as a variety of odd-numbered and branched-chain fatty acids. The fatty acid composition of neutral glycerides, largely alkyldiacylglycerols, from heat-dried ovaries of C. crispatus was not materially different from that of alkyldiacylglycerols isolated from the ovaries of frozen individuals. The fatty acid composition of neutral glycerides, largely alkyldiacylglycerols, from heat-dried ovaries of P. militaris was similar to that for C. crispatus in that 20:1(n-13) and 18:1(n-7) were major components, and odd-numbered together with branched-chain fatty acids accounted for about 15% of the total fatty acids. The fatty acid composition of neutral glycerides, consisting of equal amounts of triacylglycerols and alkyldiacylglycerols, from heat-dried ovaries of A. lincki differed from those of the previous two species in that 20:1(n-9) and 18:1(n-9) were major components and odd-numbered together with branched-chain fatty acids accounted for less than 5% of the total fatty acids. The results are discussed mainly in terms of lipid biosynthetic mechanisms within the asteroids and the interplay between internal biosynthetic activities and dietary fatty acids, particularly those from bacterial sources, is considered.     

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.     

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.     

Lipids of some Caribbean and Red Sea corals: total lipid,wax esters,triglycerides and fatty acids

The Caribbean reef-building corals Porites porites (Pallas) and Montastrea annularis (Ellis and Solander) and the Red Sea corals Pocillopora verrucosa (Ellis and Solander), Stylophora pistillata (Esper) and Goniastrea retiformis (Lamark) were analysed for total lipid, wax ester and triglyceride content, and fatty acid composition. M. annularis contained about 32% of dry weight as total lipid, whereas much lower values of between 11 and 17% were recorded for the other four species. It is concluded that there is greater variation in coral total lipid than hitherto thought. The total lipid contained a substantial proportion of wax ester (22 to 49%) and triglyceride (18 to 37%). The storage lipids (wax esters and triglycerides) accounted for between 6 and 20% of the dry weight and between 46 and 73% of the total lipid. Variation in lipid content between species could not be attributed to geographical location, but the low values for total lipid in Red Sea corals may in part be due to environmental factors as these samples were collected in winter. All corals analysed contained high levels of saturated fatty acids, the most abundant fatty acids being 16:0, 18:0 and 18:1n-9. Marked differences were observed in polyunsaturated fatty acid (PUFA) content between species, with comparatively low levels of 10 and 11% of fatty acids being recorded in M. annularis and G. retiformis, respectively. The values for the other species ranged between 21 and 37%. Fatty acid composition may vary according to the proportions of fatty acids obtained from diet, algal photosynthesis and synthesis by the animal tissues.     

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.     

Potential dietary effects on the fatty acid composition of the common jellyfish Aurelia aurita

Fatty acid composition of the natural and aquarium-reared common jellyfish Aurelia aurita was investigated. Fatty acid composition of the aquarium-reared A. aurita clearly reflected that of the diet, brine shrimp (Artemia). In the same way, fatty acid composition of the natural A. aurita was assumed to reflect those of natural diets. Samples of natural A. aurita were collected from April 1995 to September 1995 in the Seto Inland Sea, Japan, and their fatty acids were analyzed by gas chromatography and mass spectrometry. Variation of fatty acid compositions was seasonal rather than dependent on body size. Two major seasonal groups were divided by the cluster analysis of the A. aurita fatty acid composition: the April–June and the August–September clusters. The April–June cluster was characterized by high contents of the (n − 3)-fatty acids of diatom origin, accumulated via the grazing food chain. By contrast, the August–September cluster was characterized by an increase in (n − 6)-fatty acids of macroalgal origin, probably transferred via the detritus food chain. These results suggest that the diet of natural A. aurita may shift between the diatom-based food chain and the detritus-based food chain. Received: 12 April 2000 / Accepted: 1 December 2000