Martin Graeve

Diet-induced changes in the fatty acid composition of Arctic herbivorous copepods: Experimental evidence of trophic markers

To verify the potential of fatty acids as trophic markers, feeding experiments were carried out with the dominant herbivorous copepods Calanus finmarchicus, C. hyperboreus and C. glacialis from the Greenland Sea during two Arctic expeditions in June/July 1991. Depending on the fatty acid composition of these copepods, the diatom Thalassiosira antarctica or the dinoflagellate Amphidinium carterae were offered as food to induce deviating fatty acid compositions. Since the copepodite Stages CV of C. finmarchicus had very low amounts of the 16:1(n−7) fatty acid but high amounts of 18:4(n−3), the specimens were fed on T. antarctica rich in 16:1(n−7) over a period of up to 42 days. At the end of the feeding experiment the portion of the 16:l(n-7) fatty acid had strongly increased by 11%, while the 18:4(n−3) fatty acid was almost depleted. In contrast, high amounts of the 16:1(n−7) fatty acid in C. hyperboreus (CV) suggested feeding on diatoms, therefore its diet was changed to A. carterae dominated by high amounts of the 18:4(n−3) fatty acid. After 47 days the portion of 18:4(n−3) increased by 8%, whereas 16:1(n−7) decreased by 3 %. In female C. glacialis the changes in the fatty acid composition after feeding with A. carterae were less pronounced as compared to the other species, due to a severe lipid loss during the experiment. The feeding experiments document the incorporation and turnover of dietary fatty acids under controlled laboratory conditions and provide clear evidence for the potential of specific fatty acids as trophic marker lipids.

Lipids in Arctic benthos: does the fatty acid and alcohol composition reflect feeding and trophic interactions?

Arctic benthic organisms of various taxa (Anthozoa, Polychaeta, Pantopoda, Crustacea, Echinodermata) were collected on the shelves off northeast Greenland, Spitsbergen and the western Barents Sea. Their fatty acid compositions were generally characterised by the predominance of the polyunsaturated fatty acids 20:5(n-3) and 22:6(n-3) together with the saturated fatty acid 16:0, which reflect the dominance of phospholipids. The fatty acid compositions of most benthic specimens were influenced by fatty acids of dietary origin. High amounts of the fatty acid 16:1(n-7), typical of diatoms, were found in different taxa from the northeast Greenland shelf. The 18:4(n-3) fatty acid, often typical of non-diatom input, was only dominant in Ophiopholis aculeata from the Spitsbergen shelf. In some taxa small amounts of wax esters were detected with alcohol moieties similar to those of the dominant Arctic copepods. The occurrence of intact wax esters, as well as the wax ester typical fatty acids 20:1(n-9) and 22:1(n-11), also suggested ingestion of large herbivorous copepods. An unusual fatty acid composition was found for most brittle stars, due to a ratio of the 18:1(n-9) and (n-7) fatty acid isomers below 1 with lowest ratios of 0.1. A similar low ratio was also detected in the polychaete Onuphis conchylega. The extremely low portions of the 18:1(n-9) fatty acid are striking, since carnivores are generally characterised by high levels of this fatty acid. A clear gradient from low 18:1(n-9) to (n-7) ratios in suspension feeders, via predatory decapods, to higher ratios in the scavenging amphipods was a major characteristic of the benthic species. Our investigations showed that lipid analyses can give important hints on trophic relationships of benthic species and may serve as means to establish the intensity of pelagic-benthic coupling.

Herbivorous or omnivorous? On the significance of lipid compositions as trophic markers in Antarctic copepods

Three dominant Antarctic copepods, Calanoides acutus, Rhincalanus gigas and Metridia gerlachei (copepodite stages V and females), were collected during summer (January/ February) in the southern Weddell Sea south of 70°S. Detailed analyses of their lipid and fatty acid/ alcohol compositions were carried out. The trophic positions of these copepods were elucidated by means of the lipid compositions (“marker lipids”). High amounts of wax esters were found in C. acutus (92% of total lipids) and in R. gigas (84–86%). The level of wax esters in M. gerlachei was relatively low (27–42%), while the accumulation of triacylglycerols tended to be higher (19–22%). Characteristic lipid components of C. acutus were the long-chain monounsaturated fatty acids and fatty alcohols 20:1 (n-9) and 22:1 (n-11). These components together with elevated amounts of the 18:4 (n-3) and, to a lesser degree, of the 16:! (n-7) fatty acids, typical of phytoplankton lipids, indicate herbivorous feeding for C. acutus. Other abundant fatty acids were 20:5 (n-3) and 22:6 (n-3). The fatty acid composition of M. gerlachei was characterized by very high amounts of these 22:6 and 20:5 acids. Other important fatty acids were 18:1 (n-9) and 16:0, but only small amounts of 16:1 (n-7) and 18:4 (n-3) occurred. In contrast to C. acutus the fatty alcohols of M. gerlachei consisted almost exclusively of the short-chain components 14:0 and 16:0 M. gerlachei is known as an omnivorous species, which was clearly reflected by its lipid and fatty acid/alcohol pattern. Few data are available on the feeding of R. gigas, but it is usuaally described as an herbivorous small-particle feeder. R. gigas showed fatty acid/alcohol characteristics typical of either C. acutus or M. gerlachei. Higher amounts of the 16:1 (n-7) and 18:4 (n-3) fatty acids suggest herbivorous feeding, whereas the dominance of short-chain alcohols (14:0 and 16:0) resembled the lipid pattern found in the omnivorous M. gerlachei. Hence, the lipid composition of R. gigas showed an intermediate pattern, which implies a tendency towards an opportunistic feeding mode, positioned somewhere between the other two species.

Ontogenetic and seasonal changes in lipid and fatty acid/alcohol compositions of the dominant Antarctic copepods Calanus propinquus, Calanoides acutus and Rhincalanus gigas

Lipid compositions of the dominant Antarctic copepods Calanoides acutus, Rhincalanus gigas and Calanus propinquus from the Weddell Sea have been investigated in great detail. Copepods were collected during summer in 1985 and late spring/early winter in 1986. The analyses revealed specific adaptations in the lipid biochemistry of these species which result in very different lipid components. The various copepodite stages of C. acutus synthesize wax esters with long-chain monounsaturated moieties and especially the alcohols consisted mainly of 20:1(n-9) and 22:1(n-11). R. gigas also generates wax esters, but with moieties of shorter chain length. The fatty alcohols consisted mainly of 14:0 and 16:0 components, while the major fatty acids were 20:5, 18:4 and 22:6, of which 18:4 probably originated from dietary input. In contrast, C. propinquus accumulates triacylglycerols, a very unusual depot lipid in polar calanoid copepods. Major fatty acids in C. propinquus were the long-chain monounsaturates 22:1(n-9) and 22:1(n-11), which may comprise up to 50% of total fatty acids. In C. acutus and C. propinquus there was a clear increase of long-chain fatty acids with increasing developmental stage. In contrast, the fatty acid and alcohol composition of the R. gigas copepodite stages were characterized by the dominance of the polyunsaturated fatty acids as well as high amounts of the monounsaturates 18:1(n-9) and 16:1(n-7). There was a considerable decrease of the dietary fatty acid 18:4(n-3) towards the older stages during summer; in late winter/early spring 18:4 was only detected in very low amounts. This tendency was also found in the other two species, but was less pronounced. In all three species dry weight and lipid content increased exponentially from younger to older stages. The highest portion of wax esters, or of triacylglycerols in C. propinquus, was found in the adults. Dry weight and lipid content were generally higher during summer. In late winter/early spring the variability was more pronounced and lipid-rich specimens showed a selective retention of long-chain monounsaturated fatty acids, whereas in lipid-poor specimens these fatty acids were very much depleted.

Improved separation and quantification of neutral and polar lipid classes by HPLC-ELSD using a monolithic silica phase: Application to exceptional marine lipids

An improved HPLC method is presented, which allows separation and quantification of a broad range of lipid classes of marine zooplankton with special regard to neutral lipids. Marine zooplankton species often produce high amounts of exceptional lipids, especially at high latitudes, in order to cope with the harsh environmental conditions and strong seasonality in food supply. Major neutral lipid classes are wax esters, triacylglycerols, diacylglycerol ethers, free fatty alcohols and sterols. Neutral and polar lipids were separated and identified on a monolithic silica column (Chromolith Performance-Si) using high performance liquid chromatography (HPLC) with an evaporative light scattering detector (ELSD). The method resolves a broad spectrum of lipids, varying in polarity from squalene to lysophosphatidylcholine in a single run. The total run time was 35 min including column re-equilibration. The calibration was made at levels of 0.1-60 microg lipid/injection, but a 10-15-fold greater amount can be injected if single lipid classes need to be separated, e.g. for further determination of individual fatty acids. The method was applied to representative Arctic zooplankton species (copepods, pteropods, euphausiids and ctenophores) that are known to biosynthesize in particular neutral lipids like diacylglycerol ethers and free fatty alcohols.

Combined lipid, fatty acid and digestive tract content analyses: a penetrating approach to estimate feeding modes of Antarctic amphipods

Abstract. Antarctic amphipods from the Weddell Sea and Bransfield Strait were collected to investigate the impact of various species and feeding types on lipid and fatty acid compositions. In combination with digestive tract content analyses, such information can help clarify the type of feeding mode of the various amphipod species. Micro- and macropredatory amphipod species had only small amounts of triacylglycerols as storage lipids, whereas the deposit-feeder Epimeria georgiana was rich in triacylglycerols (55% of total lipids). The fatty acids 22:6(n-3), 20:5(n-3), 18:1(n-9) and 16:0 were major lipid components of most species. Ampelisca richardsoni, a suspension feeder, had a high amount of 18:4(n-3), a major component of cryptophytes and/or haptophytes, connected with feeding on sedimenting phytoplanktonic material and with a strong bentho-pelagic coupling. In Oradarea edentata, fragments of brown algae were found almost exclusively. The major fatty acid of the macroalgae, 20:4(n-6), replaced the 22:6(n-3) in the phospholipids and triacylglycerols of the amphipod. The sponge eater, Echiniphimedia hodgsoni, was rich in 16:1(n-7) and 18:1(n-7), suggesting that the unidentifiable organic matter was of diatom origin. Eusirus perdentatus, a typical predator, had high proportions of saturated and monounsaturated fatty acids, showing no specialisation in lipid and fatty acid composition. The fatty acid composition of Epimeria georgiana was similar to that of Eusirus perdentatus. However, high levels of triacylglycerols in Epimeria georgiana reflect periodical food plenty and starvation, due to its dependence on dead items. The fatty acid composition of the necrophage Waldeckia obesa was clearly different because of the predominance of 18:1(n-9) (>40% of total fatty acids). This dominance is probably the result of feeding on highly degraded carrion-derived organic matter, which is the major food of W. obesa.

Fatty acid and alcohol composition of the small polar copepods, Oithona and Oncaea : indication on feeding modes

The fatty acid and alcohol compositions of the Antarctic copepods Oithona similis, Oncaea curvata, Oncaea antarctica and the Arctic Oncaea borealis were determined to provide the first data on their lipid biochemistry and to expand the present knowledge on their feeding modes and life-cycle strategies. All these tiny species contained high amounts of wax esters (on average 51.4–86.3% of total lipid), except females of Oithona similis (15.2%). The fatty-acid composition was clearly dominated by 18:1(n-9), especially in the wax-ester-rich Oncaea curvata (79.7% of total fatty acids). In all species, 16:0 and the polyunsaturated fatty acids 20:5(n-3) and 22:6(n-3), which are structural components of all membranes, occurred in significant proportions. The dominant fatty alcohols were 14:0 and 16:0. In Oncaea antarctica and Oncaea borealis, the 20:1(n-9) and 22:1(n-11) alcohols and, to a lesser extent, the corresponding fatty acids were also found in high proportions. This indicates carnivorous feeding, although de novo biosynthesis cannot be excluded. The variable composition might be due to a wider range of food items and parasitic feeding. Typical trophic marker fatty acids for phytoplankton ingestion occurred only in small amounts, which suggests that the species were feeding on particles such as detritus or aggregates and not on living phytoplankton. From the compositional data of fatty acids and alcohols, it can be concluded that feeding behaviour of all species is omnivorous and/or carnivorous.

Lipid, fatty acid and protein utilization during lecithotrophic larval development of Lithodes santolla (Molina) and Paralomis granulosa (Jacquinot)

During the larval development of the subantarctic king crab, Lithodes santolla, and stone crab, Paralomis granulosa, we compared changes in the carbon, fatty acid and protein contents of larvae reared under constant conditions from hatching to metamorphosis, either in presence or absence of food (Artemia spp. nauplii). In both species the feeding condition had no influence on any of the chemical parameters studied, indicating a fully lecithotrophic (i.e. non-feeding) mode of development from hatching of the first zoea to metamorphosis of the late megalopa. Dry mass and carbon contents at hatching were similar in the larvae of both species, but L. santolla contained initially higher total amounts of fatty acids and protein than P. granulosa. Both species utilized considerable portions of their total fatty acid pool which decreased logarithmically throughout the time of development. At metamorphosis, it was almost exhausted in P. granulosa, while L. santolla had consumed only about 60%. Protein utilization, in contrast, was higher in L. santolla (40%) than in P. granulosa (20%). Triacylglycerol was the principal storage lipid in both species, accounting initially for about 75% of the lipid fraction; it was strongly utilized during larval development. Phospholipid constituted the second largest lipid class; it also decreased in P. granulosa, but to a lesser extent in L. santolla. The major fatty acids of both species were 18:1(n9), 20:5(n3) and

Perspectives on marine zooplankton lipids

We developed new perspectives to identify important questions and to propose approaches for future research on marine food web lipids. They were related to (i) structure and function of lipids, (ii) lipid changes during critical life phases, (iii) trophic marker lipids, and (iv) potential impact of climate change. The first addresses the role of lipids in membranes, storage lipids, and buoyancy with the following key question: How are the properties of membranes and deposits affected by the various types of lipids? The second deals with the importance of various types of lipids during reproduction, development, and resting phases and addresses the role of the different storage lipids during growth and dormancy. The third relates to trophic marker lipids, which are an important tool to follow lipid and energy transfer through the food web. The central question is how can fatty acids be used to identify and quantify food web relationships? With the fourth, hypotheses are presented on effects of global warmi...

On the lipid biochemistry of polar copepods: compositional differences in the Antarctic calanoids Euchaeta antarctica and Euchirella rostromagna

During austral summer of 1985 different developmental stages (CIII, CIV, CV, females, males) of the Antarctic copepod Euchaeta antarctica and females of Euchirella rostromagna were collected in the southeastern Weddell Sea to determine their lipid contents and compositions. For E. antarctica the analyses revealed a strong ontogenetic accumulation of lipids towards the older copepodids with highest lipid contents in late CV stages and adults. The females of E. rostromagna had moderate lipid levels. The most striking difference between these two species concerns their lipid class compositions. E. antarctica deposited predominantly wax esters, whereas in E. rostromagna the major lipid class consisted of triacylglycerols, an unusual storage lipid in polar marine copepods. Principal fatty acids in E. antarctica were the monounsaturates 18:1(n-9) and 16:1(n-7), especially in the lipid-rich stages, while the polyunsaturated fatty acids 20:5(n-3) and 22:6(n-3), usually membrane lipids, dominated in the lipid-poor stages. The wax ester moieties in E. antarctica consisted almost entirely of 14:0 and 16:0 fatty alcohols. Major components in E. rostromagna were the fatty acids 18:1(n-9), 16:0, 20:5(n-3) and 22:6(n-3). The potential of fatty acids and alcohols as typical trophic markers is rendered largely insignificant in the two species due to catabolic processes.