Ontogenic variations in fatty acid and alcohol composition of the pelagic amphipod <Emphasis Type="Italic">Themisto libellula</Emphasis> in Kongsfjorden (Svalbard)

The fatty acid and alcohol composition of the pelagic amphipod, Themisto libellula, was monitored during the 5 first months of its life cycle (4–20 mm length) in an Arctic fjord, Kongsfjorden, Svalbard. Fatty acids of the three major lipid classes, polar lipids (PL), triacylglycerol (TAG), and wax esters (WE), were analyzed to highlight ontogenic changes in their diet and metabolism. The PL composition of T. libellula did not show any strong variations along their growth except during the first month where an important increase of 20:5(n-3) (EPA) and 22:6(n-3) (DHA) was observed. The TAG composition revealed a clear gradient corresponding to a diet shift from omnivorous juveniles toward carnivorous sub-adults and adults. Indeed, fatty acid trophic markers of diatoms were dominant in the juveniles, whereas 20:1(n-9) and 22:1(n-11), the Calanus sp. trophic markers, overwhelmed in the older stages. The WE composition highlighted the same general trend, however, differences were found with the TAG and are discussed as a result of differences in turnover rates and assimilation pathways between the two lipid classes.     

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.     

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     

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

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

Trophic relationships of white-chinned petrels from Crozet Islands: combined stomach oil and conventional dietary analyses

The diet of white-chinned petrels Procellaria aequinoctialis breeding at the Crozet Archipelago (southern Indian Ocean) was studied using two complementary methods: lipid analysis of stomach oils as trophic markers together with the conventional dietary approach (i.e., stomach content analysis). Objectives were (1) to investigate the adult diet when they feed for themselves by analyzing stomach oil lipids, and (2) to compare the lipid signature of chick and adult oils. Stomach oils mainly consisted of triacylglycerols (TAG), diacylglycerol-ethers (DAGE) and wax esters (WE) (66, 14 and 11%, respectively). The dietary origin of TAG and WE was evaluated by linear discriminant analyses with fatty acid and fatty alcohol fractions. Analyses evidenced that stomach oils did not originate from Antarctic krill, but instead from myctophid fish, thus demonstrating the importance of mesopelagic fish in the nutrition of adult petrels. This result was consistent with the identification of digested remains of myctophids recovered from adult stomach contents after long foraging trips. Large amounts of a rare lipid class, DAGE (up to 76% of total lipids), were identified in two stomach oils, together with fresh remains of the squid Gonatus antarcticus (99% by mass), suggesting that DAGE could have the potential to be trophic markers of cephalopods. Moreover, six oils probably originated from Patagonian toothfish, thus confirming strong interactions between white-chinned petrels and fisheries. Comparison between chick and adult stomach oils indicated no major differences in their biochemical composition suggesting an identical dietary origin of oils, mainly myctophids. Both adult and chick oils can therefore be used to determine the feeding ecology of adult birds when they feed far away from their breeding grounds. Finally, food analysis of chick samples and adult samples collected after short and long trips indicated different foraging grounds during the two kinds of trips, and also between long trips performed in subtropical and Antarctic waters.     

Association of <Emphasis Type="Italic">Waminoa</Emphasis> sp. (Acoela) with corals in the Wakatobi Marine Park,South-East Sulawesi,Indonesia

This is the first quantitative study on the prevalence of epizoic Waminoa sp. acoel worms and their association with corals in the Wakatobi Marine National Park (WMNP), South-East Sulawesi, Indonesia. Three replicate transects were laid on the reef crest, flat and slope at six sites in 2006 and eight sites in 2007. Four of the sites were common in both years. In total 69 transects were surveyed in 2006, and 87 transects in 2007. A total of 4.8% of all observed hard corals were associated with acoel worms in 2006 and 2.6% of hard and soft corals in 2007. Acoels were present on 16 and 21 of the coral taxa studied in 2006 and 2007 respectively. The worms were strongly associated with the azooxanthellate coral Tubastrea spp. and were rare or absent on the most abundant coral genera Montipora and Porites. The mean number of corals having acoels was highest on reef slopes, whereas acoels were virtually absent on reef flats. Corals that had a high and a medium cover of worms were more common in 2007 than in 2006. No significant trend in the adaptation of the zooxanthellae of Waminoa sp. to different depths at different sites was revealed. The impact of the worm on the coral is unknown, but high numbers may have a shading effect and a negative impact on the coral’s photophysiology. This acoel merits more study of its life cycle, its photophysiology, and its impact on its host corals. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Changes in yolk total proteins and lipid components and embryonic growth rates during lobster (<Emphasis Type="Italic">Homarus americanus</Emphasis>) egg development under a simulated seasonal temperature cycle

From August 2000 to June 2001, seven egg-carrying female lobster (Homarus americanus) from the Îles de la Madeleine population (Gulf of St. Lawrence, Canada) were held under a simulated seasonal temperature cycle to monitor egg development from extrusion to hatching. For the first time, changes in the yolk components (total lipids and major lipid classes, total proteins) and embryo growth of single eggs were monitored separately over the entire development period. Under the controlled temperature conditions, egg development proceeded in three phases. (1) Autumn, from extrusion to early December, was marked by a rapid increase in the Perkinss eye index and rapid declines in yolk total proteins and triacylglycerols (TAG). Embryo daily growth rate was estimated between 1 and 2 µg proteins day^–1. (2) Winter, from late December to early April (temperature stable at ca. 1°C) was characterized by a stationary phase in the evolution of the eye index and yolk lipid use, and embryo growth slowed significantly. (3) Spring, from late April to hatching in June was the period with the most rapid changes in yolk TAG and embryo growth rates >6 µg proteins day^–1 were recorded. Almost 65% of the live biomass (total proteins) of the hatching larvae was accumulated during the last few weeks of development. An index of embryo growth efficiency was estimated as the slope of the relationship between embryo total proteins and yolk TAG during egg development. A relationship was found between the initial mean egg dry weight and the embryo growth efficiency index suggesting that under the same experimental conditions bigger eggs used yolk lipids more efficiently and sustained faster embryonic growth than smaller eggs. The relationship may also explain why larger larvae originate from larger eggs.Communicated by R.J.Thompson, St Johns     

Effect of additions of dietary triacylglycerol microspheres on growth,survival, and settlement of mussel (<Emphasis Type="Italic">Mytilus</Emphasis> sp.) larvae

This study reports the effect of additions of dietary microspheres of triacylglycerol (TAG, extracted from the diatom Chaetoceros muelleri) on larval development and settlement of Mytilus sp. The first experiment showed that mussel larvae successfully ingested TAG microspheres as soon as they acquired the ability to feed from exogenous sources. In a second experiment, larvae were fed for 28 days on diets consisting of 0, 1, 20, or 50% TAG microspheres (based on the cell concentration of a full algal ration) added as partial replacements for a ration of Isochrysis sp. (T-ISO). Lipid content and growth of larvae fed on a diet composed of 20% TAG were higher than those of the control groups, whereas survival was negatively affected. No growth or survival effect was detected with larvae fed on a diet composed of 1% TAG, whereas high mortality after 14 days was observed for larvae fed on a diet composed of 50% TAG microspheres. In a third experiment, 22-day-old larvae were fed on rations of Isochrysis sp. supplemented with 0, 1, 10, 20, 50, and 100% TAG for 2 days and allowed to settle for a 7-day period. TAG content of the larvae increased with TAG added to the diet until a saturation threshold was reached between a 20 and 50% supplementation level. Similarly, free fatty acid (FFA) content increased with TAG level in the diet and was linearly correlated with TAG content of larvae. Increased levels of FFA in larvae were attributed to digestion of TAG supplements. Settlement success and survival of larvae were not affected by diet; however, regression analysis revealed that TAG level in pre-metamorphic larvae explained 28% of survival variability among cultures.Communicated by R.J. Thompson, St. Johns     

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.     

The fate of dietary lipids in the Arctic ctenophore Mertensia ovum (Fabricius 1780)

Lipids of the Arctic ctenophore Mertensia ovum, collected from Kongsfjorden (Svalbard) in 2001, were analysed to investigate seasonal variability and fate of dietary lipids. Total lipids, lipid classes and fatty acid and alcohol compositions were determined in animals, which were selected according to age-group and season. Changes in lipids of age-group 0 animals were followed during growth from spring to autumn. Total lipids increased from May to September. Lipids as percentage of dry mass were lowest in August indicating their use for reproduction. Higher values occurred in September, which may be due to lipid storage for overwintering. Wax esters were the major lipid class accounting for about 50% of total lipids in age-group 0 animals from July and August. Phospholipids were the second largest lipid fraction with up to 46% in this age-group. The principal fatty acids of M. ovum from all age-groups were 22:6(n-3), 20:5(n-3) and 16:0. Wax ester fatty alcohols were dominated by 22:1(n-11) and 20:1(n-9) followed by moderate proportions of 16:0. The unique feature of M. ovum lipids was the high amount of free fatty alcohols originating probably from the dietary wax esters. In May, free alcohols exhibited the highest mean proportion with 14.6% in age-group 0 animals. We present the first data describing a detailed free fatty alcohol composition in zooplankton. This composition was very different from the alcohol composition of M. ovum wax esters because of the predominance of the long-chain monounsaturated 22:1(n-11) alcohol accounting for almost 100% of total free alcohols in some samples. The detailed lipid composition clearly reflected feeding of M. ovum on the herbivorous calanoid species, Calanus glacialis and C. finmarchicus, the abundant members of the zooplankton community in Kongsfjorden. Other copepod species or prey items seem to be less important for M. ovum.     

Swimming ability of eels (<Emphasis Type="Italic">Anguilla rostrata,Conger oceanicus</Emphasis>) at estuarine ingress: contrasting patterns of cross-shelf transport?

The transport of eel early life stages may be critical to their population dynamics. This transport from ocean spawning to freshwater, estuarine and coastal nursery areas is a combination of physical and biological processes (including swimming behavior). In New Jersey, USA, the American eel (Anguilla rostrata) enters estuaries as glass eels (48.7–68.1 mm TL) in contrast to the Conger eel (Conger oceanicus) that enters as larger (metamorphosing) leptocephali (68.3–117.8 mm TL). To begin to understand the mechanisms of cross-shelf transport for these species, we measured the potential swimming capability (critical swimming speed, U _crit) under ambient conditions throughout the ingress season. A. rostrata glass eels were collected over many months (January–June) at a range of temperatures (4–21°C), with relative condition declining over the course of the ingress period as temperatures warmed. C. oceanicus occurred later in the season (April–June) and at warmer temperatures (14–24.5°C). Mean U _crit values for A. rostrata (11.7–13.3 cm s⁻¹) and C. oceanicus (14.7–18.6 cm s⁻¹) were comparable, but variable, with portions of the variability explained by water temperature, relative condition, ontogenetic stage, and fish length. Travel times to Little Egg Inlet, New Jersey, estimated using 50% U _crit values, indicate it would take A. rostrata ~30 and ~60 days to swim from the shelf edge and Gulf Stream, respectively. Travel times for C. oceanicus were shorter, ~20 days from the shelf edge, and ~45 days from the Gulf Stream. Despite differences in life stage, our results indicate both species are competent swimmers, and suggest they are capable of swimming from the Gulf Stream and/or edge of the continental shelf to estuarine inlets.     

Temporal genetic variation in populations of the limpet Cellana grata from Hong Kong shores

Variations in the relative contributions of gene flow and spatial and temporal variation in recruitment are considered the major determinants of population genetic structure in marine organisms. Such variation can be assessed through repeated measures of the genetic structure of a species over time. To test the relative importance of these two phenomena, temporal variation in genetic composition was measured in the limpet Cellana grata, among four annual cohorts over 10 years at four rocky shores in Hong Kong. A total of 408 limpets, comprising individuals from 1998, 1999, 2006 and 2007 cohorts were screened for genetic variation using five microsatellite loci. Minor but significant genetic differentiation was detected among samples from the 1998/1999 collection (F _ST = 0.0023), but there was no significant differentiation among the 2006/2007 collection (F _ST = 0.0008). Partitioning of genetic variation among shores was also significant in 1998/1999 but not in the 2006/2007 collection, although there was no correlation between genetic and geographic distances. There was no significant difference between collections made in 1998/1999 and 2006/2007. This lack of clear structure implies a high level of gene flow, but differentiation with time may be the result of stochastic recruitment variation among shores. Estimates of effective population size were not high (599, 95% C.L. 352–11397), suggesting the potential susceptibility of the populations to genetic drift, although a significant bottleneck effect was not detected. These findings indicate that genetic structuring between populations of C. grata in space and time may result from spatio-temporal variation in recruitment, but the potential development of biologically significant differentiation is suppressed by a lack of consistency in recruitment variability and high connectivity among shores.     

Variability in nursery function of tropical seagrass beds during fish ontogeny: timing of ontogenetic habitat shift

Seagrass beds are often considered to be important nurseries for coral reef fish, yet the effectiveness of these nursery functions (refuge and food availability) at different juvenile stages is poorly understood. To understand how the demands of juvenile fish on seagrass nursery functions determines the timing of ontogenetic habitat shifts from seagrass beds to coral reefs, we conducted visual transect survey and field tethering and caging experiments on three different sizes of the coral reef fish Pacific yellowtail emperor (Lethrinus atkinsoni) during its juvenile tenure in seagrass beds at Ishigaki Island, southern Japan. The study showed that although the number of individual L. atkinsoni juveniles decreased by >90 % during their stay in the seagrass nursery, the shelter and/or food availability functions of the nursery, at least for a juvenile size of approximately 5 cm total length (TL), provided the best survival and growth option. The timing of ontogenetic migration to coral reefs of larger fish (>8 cm TL) was attributed to foraging efficiency for larger food items in different habitats. Overall, the function of the seagrass bed nursery changed with juvenile body size, with marginally higher survival and significantly greater growth rates during early juvenile stages in seagrass beds compared to coral reefs. This would contribute to the enhancement in the number of individuals eventually recruited to adult populations.     

Laboratory culture of the aeolid nudibranch <Emphasis Type="Italic">Spurilla neapolitana</Emphasis> (Mollusca,Opisthobranchia): life history aspects

Spurilla neapolitana from the Eastern Mediterranean was cultured in a self-sustained, temperature-controlled laboratory culture system, and its life cycle is described. Adults were collected from three field sites situated 120 km apart, along the Israeli Mediterranean coastline, between March 2006 and August 2007. Cultures of the life-cycle stages were raised at 24°C. S. neapolitana deposited white, coiled, spiral egg masses containing zygotes. Veliger larvae hatched 3.0 ± 0.4 days post oviposition. The veliger larvae are obligatory planktotrophs, with a minimal larval phase of 22 days. In the lab, larvae settled and metamorphosed following exposure to metabolites derived from distinct prey sea anemone species. Reproductive maturity was reached 42 ± 5 days post metamorphosis, resulting in a laboratory generation time of 67 days (egg to egg). The average life span of reproductive specimens in this study was 157 ± 13 days post-oviposition and they reached a length of 7–10 cm. During this period, an average adult deposited ca. 40 × 10⁶ zygotes. This species has several characteristics that suggest it will be a useful model for laboratory-oriented research. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Proximate composition and nucleic acid content of premetamorphic leptocephalus larvae of the congrid eel Ariosoma balearicum

Water, ash, proximate composition (protein, lipid, carbohydrate, hexosamine), and nucleic acid (DNA, RNA) content were measured in premetamorphic larvae of the congrid eel Ariosoma balearicum (Delaroche) collected from the eastern Gulf of Mexico. Specimens ranged from 15.0 to 202.3 mm total length (TL) and 0.0116 to 4.3860 g wet mass (WM). Water content increased linearly with increasing specimen mass over the entire size range; consequently, percent water was uniform and had a mean value of 92.9±1.09% WM. Ash content also increased linearly with specimen mass, but only up to a mass of 2.5 g WM (165 mm TL). Ash content in specimens >165 mm TL showed only a small increase with mass, suggesting an improved osmoregulatory capability in larger individuals. The absolute amount of all proximate components increased with increasing specimen size, but rates of deposition among the components varied, resulting in different patterns in the relative concentrations of each with growth. Protein dominated the ash-free dry mass (AFDM) throughout development (29 to 59% AFDM); carbohydrate and hexosamine occurred in similar proportions (8 to 24% AFDM). Lipid was a significant proportion of the AFDM in only the smallest individuals. Lipid concentrations decreased initially as mass increased in individuals smaller than 0.4 g WM (90 mm TL), indicating a low rate of lipid deposition in small individuals. In specimens >90 mm TL, lipid concentrations were uniform and had a mean of 12% AFDM. Trends for biochemical components and nucleic acids suggest that growth of Phase I leptocephali occurs in two subphases (Ia and Ib). Phase Ia is characterized by cellular proliferation, preferential synthesis of protein and carbohydrate relative to lipid, and growth manifested more as increased length rather than increased mass. For A. balearicum, Phase Ia extends from yolk-sac absorption to 90 mm TL. In Phase Ib, nucleic acid content levels off, lipid deposition increases, and mass increases exponentially.     

Patterns of growth and triacylglycerol content in snow crab Chionoecetes opilio (Brachyura: Majidae) zoeal stages reared in the laboratory

Snow carb Chionoecetes opilio zoea I and zoea II larvae, hatched from females in a controlled mating experiment, were reared in the laboratory at 10.1 °C and 28.0 salinity, to resolve the patterns of growth (dry weight [DW]) and change in energy reserves (triacylglycerols [TAG]) within a given moult cycle. The patterns of growth and change in TAG reserves were similar in each zoeal stage. Following hatching or a moult, the zoeae entered a phase of rapid size increase, i.e. high daily growth rates (5.5 to 12.8% DWd^-1), for 1/3 to 2/5 of the duration of the moult cycle. During the same period, the zoeae accumulated TAG reserves until a maximum (TAG DW^-1) was reached at the end of the phase of rapid growth. The period of high growth rates and of TAG accumulation is interpreted as the required time for the zoeae to reach a point in development [i.e. point of reserves saturation (PRS); Anger and Dawirs (1981)] where sufficient growth and energy reserves allow moulting to the next stage. Following the phase of rapid growth and TAG accumulation, the zoeae entered a phase of low daily growth rates (0 to 1% DWd^-1) during which the TAG reserves decreased to a minimum at the end of the phase. Prior to, and during the moult to zoea II, a phase of negative growth was observed in the zoea I larvae. We conclude that measurement of zoeal size and TAG content, along with morphometric criteria (e.g. epidermal retraction), can be used to assess growth and nutritional condition of C. opilio zoeal stages from the sea.     

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     

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.     

Ontogeny of space use and diet of two temperate damselfish species, <Emphasis Type="Italic">Parma microlepis</Emphasis> and <Emphasis Type="Italic">Parma unifasciata</Emphasis>

Studies of reef fish herbivory have mainly focused on the impacts and behaviour of adults of tropical species. In this study, the ontogenetic shifts in home range, aggression, feeding rate, diet and gut morphology in juveniles and adults of two temperate territorial damselfishes, Parma microlepis and Parma unifasciata, were determined. Both P. microlepis and P. unifasciata juveniles under 80 mm TL exhibited no aggressive chases towards conspecifics or other species, while above 80 mm TL aggressive chase frequency increased in conjunction with an increase in home range, defended as a territory. Ontogenetic diet shifts, characterised by an increase in herbivory (P. unifasciata: juveniles: 64% plant material, adults: 95% plant material; P. microlepis: juveniles: 43% plant material, adults: 67% plant material) were observed for both species. The ratio of digestive tract length to body length, which often accompanies a switch to herbivory, increased significantly with ontogeny for both species. Compared to tropical confamilial grazers, these temperate damselfish species feeding rates were lower, and they had larger territories which were not as strongly defended (fewer aggressive chases).     

Neutral lipid compositions of antarctic fish tissues may reflect use of fatty acyl substrates by catabolic systems

Antarctic fishes contain large quantities of lipid in adipose tissue, blood and oxidative muscle. In this study, the fatty acyl compositions of free fatty acids (FFA) and triacylglycerol (TAG) in serum, adipose tissue and oxidative muscle were determined in two species of Antarctic fishes, Trematomus newnesi Boulenger, 1902, and Notothenia gibberifrons Lönnberg, 1905, collected off the Antarctic Peninsula in February and March 1987. Total lipid contents of serum and oxidative muscle in each species also were measured. T. newnesi, a mesopelagic species, has significantly more lipid in the serum than N. gibberifrons, a demersal species (17.6±2.4 and 10.4±1.0 mg lipid g^-1 serum, respectively). The oxidative muscle of T. newnesi also contains more lipid than does that of N. gibberifrons (12.5 and 9.3% by dry weight, respectively). Fatty acids comprising greater than 5% of one or more of the lipid pools assayed in both species include 14:0, 16:0, 16:1, 16:2, 18:0, 18:1, 20:5 and 22:6. The percentages of 16:1 were lower (p<0.05) in oxidative muscle FFA than in all other assayed lipid pools in both species, suggesting a catabolic preference for some monoenes in oxidative muscle of these fishes. The percentage of 14:0 was two-to three-fold higher in adipose tissue TAG than in all other lipid pools in both species. Differences in fatty acid compositions between lipid pools indicate than proteins involved in the mobilization, transport or catabolism of fatty acids exhibit differential recognition of fatty acids in T. newnesi and N. gibberifrons.