Fatty acids of sponges from the Sea of Okhotsk

The fatty acid (FA) composition of Demospongiae species from the Sea of Okhotsk was studied. Fifteen sponge species were investigated for the first time, and the previously studied species Desmacella rosea and Myxilla incrustans were reexamined for their FA composition. Gas chromatography-mass spectrometry revealed 150 different fatty acids, of which 15 have not been identified in sponge lipids previously. The relative content of saturated FAs varied from 7.6 in Melonachora kobjakovae to 29.6% in Amphilectus digitata, with an average of 14.6% of total FAs. The relative content of monoenic FAs ranged from 12.8 in T. dirhaphis to 27.0% in Polymastia sp., with an average of 20.6% of total FAs. Non-methylen-interrupted, primarily unsaturated Δ5,9-FAs contributed a significantly to the amount of polyunsaturated fatty acids of sponges; this being a distinguishing feature of the FA composition of the investigated group of organisms.     

Lipid class and fatty acid compositions of the zoanthid Palythoa caesia (Anthozoa: Hexacorallia: Zoanthidea) and its chemotaxonomic relations with corals

Lipid class and fatty acid (FA) compositions of the zoanthid Palythoa caesia from the South China Sea (Vietnam) were determined and compared with those of zooxanthellate reef-building and soft corals from the same region to clarify chemotaxonomic relations between zoanthids and corals. Total lipids of P. caesia and corals were formed by the same lipid classes. The lipids of P. caesia contained specific for cnidarians monoalkyldiacylglycerols and phosphonolipids. The proportion between reserve and structural lipid classes of P. caesia was similar to that of alcyonarians (soft corals), but significantly differed from reef-building corals. This similarity may be caused by the similar energy budget of cnidarian colonies without hard exoskeleton. The distribution of polyunsaturated FAs (PUFAs), such as 22:4n-6 and 22:5n-3, indicated pathways of biosynthesis of PUFAs in Palythoa to be closer to those in reef-building corals than in soft corals. The differences in the profiles of common FAs between P. caesia and reef-building corals were showed. Zoanthid lipids contained rare Δ5,9 non-methylene-interrupted (NMI) PUFAs (with two and three double bounds), which probably originated from microorganisms associated with P. caesia. Trienoic acids Δ5,9,15–24:3 and Δ5,9,17–24:3 were found in Palythoa for the first time. NMI PUFAs, as well as total FA profile, can be used for chemotaxonomy of Palythoa.     

Biosynthesis of long-chain polyunsaturated fatty acids in the razor clam Sinonovacula constricta: Characterization of four fatty acyl elongases and a novel desaturase capacity

As an unusual economically important aquaculture species, Sinonovacula constricta possesses high levels of long-chain polyunsaturated fatty acids (LC-PUFA). Previously, our group identified fatty acyl desaturases (Fad) with Δ5 and Δ6 activities in S. constricta, which was the first report of Δ6 Fad in a marine mollusc. Here, we further successfully characterize elongases of very long-chain fatty acids (Elovl) in this important bivalve species, including one Elovl2/5, two Elovl4 isoforms (a and b) and a novel Elovl (c) with Elovl4 activity. In addition, we also determined the desaturation activity of S. constricta Δ6 Fad toward 24:5n-3 to give 24:6n-3, a key intermediate in docosahexaenoic acid (DHA) biosynthesis. Therefore, S. constricta is the first marine mollusc reported to possess all Fad and Elovl activities required for LC-PUFA biosynthesis via the ‘Sprecher pathway’. This finding greatly increases our understanding of LC-PUFA biosynthesis in marine molluscs. Phylogenetic analysis by interrogating six marine molluscan genomes, and previously functionally characterized Elovl and Fad from marine molluscs, suggested that DHA biosynthetic ability was limited to a few species, due to the general lack of Δ4 or Δ6 Fad in most molluscs.     

New advances in fatty acids as antimalarial, antimycobacterial and antifungal agents

This review deals with the most recent findings on the antimalarial, antimycobacterial, and antifungal properties of fatty acids, with particular emphasis on novel marine fatty acids. The first section deals with the most recent and some background literature on what has been the latest developments with respect to fatty acids as antimalarial agents and the importance of enzyme inhibition, in particular the inhibition of the enoyl-ACP reductase (FabI) of Plasmodium falciparum, the principal agent responsible for malaria. This section of the review also emphasizes the latest antimalarial research with the very long-chain Delta5,9 fatty acids from sponges. The second section of the review deals with the recent literature on the antimycobacterial activity of fatty acids and the importance of enzyme inhibition, in particular the inhibition of the enoyl-ACP reductase (InhA) of Mycobacterium tuberculosis for antimycobacterial activity. The inhibitory activities of the Delta5,9 fatty acids against InhA as well as that of the alpha-methoxylated fatty acids are also discussed. The importance of Delta5,9 fatty acids as topoisomerase I inhibitors and its connection to cancer is also reviewed. The last part of the review, the antifungal section, also emphasizes the most recent research with antifungal fatty acids and the importance of enzyme inhibition, in particular N-myristoyltransferase (NMT) inhibition, for antifungal activity. This last section of the review emphasizes the latest research with the alpha-methoxylated fatty acids but the importance of acetylenic fatty acids is also considered.     

Comparative study of the endemic freshwater fauna of Lake Baikal—VI. Unusual fatty acid and lipid composition of the endemic sponge Lubomirskia baicalensis and its amphipod crustacean parasite Brandtia (Spinacanthus) parasitica

Lipids and phospholipids (both plasmalogen and alkyl forms) of the freshwater sponge Lubomirskia baicalensis and the sponge's gammarid parasite Brandtia (Spinacanthus) parasitica were examined. Composition of alkenyl-acyl (plasmalogen), alkylacyl and diacyl forms of major phospholipid classes, phosphatidylethanolamine and phosphatidylcholine were determined. One hundred and eighty-three fatty acids were identified by GC-MS: 46 saturated, 55 monoenoic, 35 dienoic, 25 trienoic and 22 tetra-, penta- and hexaenoic. The freshwater sponges, belonging to the family Lubomirskiidae, were shown to contain unusual long-chain fatty acids: anteiso-5, 9–28:2, branched-5, 9–29:2, 5,9,23–29:3, 5,9,23–30:3, 15,18,21,24–30:4 and 15,18,21,24,27–30:5. Some from these fatty acids were found in lipids of the amphipod parasite.     

A test of the Menge-Sutherland model of community organization in a tropical rocky intertidal food web

Summary Menge and Sutherland (1976) predicted that in physically benign habitats: (1) community structure will be most strongly affected be predation, (2) the effect of predation will increase with a decrease in trophic position in the food web, (3) trophically intermediate species will be influenced by both predation and competition, and (4) competition will occur among prey species which successfully escape consumers. These predictions were tested in a tropical rocky intertidal community on the Pacific coast of Panama. The most abundant mobile species included fishes and crabs, which occupied the top trophic level, and predaceous gastropods and herbivorous molluscs, which occupied intermediate trophic levels. The most abundant sessile organisms were encrusting algae, foliose algae, barnacles, and bivalves. Diets were broad and overlapping, and 30.3% of the consumers were omnivorous. Each consumer group had strong effects on prey occurring at lower trophic levels: (1) Fishes and crabs reduced the abundance of predaceous snails, herbivorous molluscs, foliose algae, and sessile invertebrates. (2) Predaceous gastropods reduced the abundance of herbivorous molluscs and sessile invertebrates. (3) Herbivorous molluscs reduced the abundance of foliose algae and young stages of sessile invertebrates, and altered relative abundances of the encrusting algae. The encrusting algae, although normally the dominant space occupiers, proved to be inferior competitors for space with other sessile organisms when consumers were experimentally excluded. However, the crusts escaped consumers by virtue of superior anti-herbivore defenses and competed for space despite intense grazing. Observations do not support the hypothesis that the trophically intermediate species compete. Hence, with the exception of this last observation, the predictions of the Menge and Sutherland model were supported. Although further work is needed to evaluate other predictions of the model in this community, evidence from this study joins an increasing body of knowledge supporting the model. Contradictory evidence also exists, however, indicating that aspects of the model require revision.     

Identification of a Δ5-like Fatty Acyl Desaturase from the Cephalopod Octopus vulgaris (Cuvier 1797) Involved in the Biosynthesis of Essential Fatty Acids

Long-chain polyunsaturated fatty acids (LC-PUFA) have been identified as essential compounds for common octopus (Octopus vulgaris), but precise dietary requirements have not been determined due, in part, to the inherent difficulties of performing feeding trials on paralarvae. Our objective is to establish the essential fatty acid (EFA) requirements for paralarval stages of the common octopus through characterisation of the enzymes of endogenous LC-PUFA biosynthetic pathways. In this study, we isolated a cDNA with high homology to fatty acyl desaturases (Fad). Functional characterisation in recombinant yeast showed that the octopus Fad exhibited Δ5-desaturation activity towards saturated and polyunsaturated fatty acyl substrates. Thus, it efficiently converted the yeast's endogenous 16:0 and 18:0 to 16:1n-11 and 18:1n-13, respectively, and desaturated exogenously added PUFA substrates 20:4n-3 and 20:3n-6 to 20:5n-3 (EPA) and 20:4n-6 (ARA), respectively. Although the Δ5 Fad enables common octopus to produce EPA and ARA, the low availability of its adequate substrates 20:4n-3 and 20:3n-6, either in the diet or by limited endogenous synthesis from C(18) PUFA, might indicate that EPA and ARA are indeed EFA for this species. Interestingly, the octopus Δ5 Fad can also participate in the biosynthesis of non-methylene-interrupted FA, PUFA that are generally uncommon in vertebrates but have been found previously in marine invertebrates, including molluscs, and now also confirmed to be present in specific tissues of common octopus.     

Effect of habitat and motor activity of molluscs on fatty acid composition of triglycerides and phospholipids

A comparative analysis of fatty acids (FA) in neutral lipids and phospholipids of digestive gland and pedal muscle has been performed in molluscs from various ecological groups differing by belonging to sea or fresh water, trophic types or the associated motor activity. In freshwater pulmonary gastropods Lymnaea stagnalis and Lymnaea ovalis and marine prosobranchial molluscs Buccinum undatum and Littorina littorea the total content of ω3-acids in phospholipids of the studied tissues differed more than twice, predominantly due to the combined effect of temperature and salinity of the habitat. The lower viscosity of cell membranes in marine species (ω3/ω6 < 1) is determined to the greatest degree by the presence of eicosapentaenoic acid that accounts for 22–25% of the FA sum in marine species. Comparison of the molluscs by their trophic belonging has revealed the presence of linoleic acid in triglycerides in digestive glands of phytophages (8–12%), but the practically complete absence of this acid in the predator B. undulatum (< 0.8%). By mobility, L. littorea inhabiting the high-low tide littoral was inferior to freshwater pulmonary gastropods and to the marine predator, as it stops moving twice a day during the low tide. In phospholipids of pedal muscle of this mollusc the amount of long-chain polyunsaturated C: 22 FA was 3–6 times lower than that in other studied species, which might possibly indicate the role of these acids in functioning of the pedal muscle contractile tissue. On the whole, use of the FA characteristics as the parameters determining belonging to certain ecological group requires a certain caution due to a complex action of biotic and abiotic factors on the animal metabolism. The exception is the ω3/ω6 ratio in total phospholipids of fresh water and marine gastropods.     

Identification and characterization of a novel C20-elongase gene from the marine microalgae, Pavlova viridis, and its use for the reconstitution of two pathways of long-chain polyunsatured fatty acids biosynthesis in Saccharomyces cerevisiae

The marine microalga, Pavlova viridis, contains long-chain polyunsatured fatty acids including eicosapentaenoic acid (EPA, 20:5n-3) and docosapentaenoic acid (DPA, 22:5n-3). A full-length cDNA sequence, pvelo5, was isolated from P. viridis. From sequence alignment, the gene was homologous to fatty acyl elongases from other organisms. Heterologous expression of pvelo5 in Saccharomyces cerevisiae confirmed that it encoded a specific C20-elongase within the n-3 and n-6 pathways. Elongation activity was confined exclusively to EPA and arachidonic acid (20:4n-6). GC analysis indicated that pvelo5 could co-express with other genes for biosynthesis to reconstitute the Δ8 and Δ6 pathways. Real-time PCR results and fatty acid analysis demonstrated that long-chain polyunsatured fatty acids production by the Δ8 pathway might be more effective than that by the Δ6 pathway.     

Effects of weaning and finishing feeding treatment on fatty acids, especially cis and trans C18:1 isomers, in the Longissimus thoracis muscle of Galician Blond calves

Seven- to 10-month-old calves are the typical production of Galician Blond (GB), the most important bovine local beef breed in Spain. As meat lipid repercussions on human health depend on their fatty acid (FA) profile, this study aimed at analysing the individual FA at weaning and at the end of the feeding finishing period, especially trans and cis 18:1 isomers in total lipids of the Longissimus thoracis muscle in GB male calves. Distribution of main FA in veal lipids was characterized by gas-liquid chromatography (GLC) analysis on a high polar glass capillary column. Total trans and cis 18:1 isomers were purified, from total FA (TFA) methyl esters, by preparative reversed-phase high-performance liquid chromatography, to allow subsequent specific analysis of their different isomers by GLC. Calves stayed with their mothers at pasture for 2 or 5 months in intensive or semi-extensive systems, followed by an indoor feeding period. The weaned (W) group was finished on concentrate and hay, whereas the non-weaned (NW) group was finished on concentrate and hay and it continued suckling until slaughter. The studied effects did not present interactions. The duration of the indoor finishing period hardly had a significant influence on the parameters analysed. Compared to W calves, NW calves had higher proportions (% TFA) of 18:3n-3 (+38%, P < 0.0001), 20:5n-3 (+22%, P = 0.005), 22:5n-3 (+13%, P = 0.042), cis monounsaturated FA (MUFA; +8%, P = 0.032), total MUFA (+8%, P = 0.013), n-3 polyunsaturated FA (PUFA; +25%, P = 0.0001) and conjugated linoleic acid (CLA; +48%, P < 0.0001) to the detriment of 18:0 (-10%, P = 0.002), 18:2n-6 (-25%, P = 0.004) and n-6 PUFA (-20%, P = 0.011). With regard to cis and trans 18:1 isomers, NW calves had higher proportions (% total cis or trans 18:1) of Δ11trans and Δ16trans (+25% and +22%, respectively, P < 0.01) and Δ13 + 14trans (+13%, P < 0.05) and lower proportions of Δ6 to 8 and Δ10trans (-28% and 58%, respectively, P < 0.001). NW calves' meat might be more suitable for human health than W calves' meat due to the increase of anti-atherogenous FA such as n-3 PUFA, CLA and 18:1 Δ11trans.     

Comparative study of the fatty acid composition of sponges of the genus Ircinia. Identification of the new 23-methyl-5,9-tetracosadienoic acid.

1. The phospholipid fatty acid compositions of the sponges Ircinia strobilina, Ircinia felix, Ircinia campana, Ircinia sp., Spongia tubulifera and Dysidea etherea were studied, revealing the presence, besides other common fatty acids, of considerable amounts (2-5%) of the novel 23-methyl-5,9-tetracosadienoic acid (1). 2. The demospongic acids 5,9-tetracosadienoic acid, 23-methyl-5,9-tetracosadienoic acid (1), and 5,9-pentacosadienoic acid, were particularly abundant in sponges of the genus Ircinia, in contrast to the most common 5,9-hexacosadienoic acid found in other species. These findings are discussed in terms of the taxonomy of the Dictyoceratida. 3. The complete characterization of the novel phospholipid fatty acid 23-methyl-5,9-tetracosadienoic acid (1) is presented.     

Effect of botanical composition of silages on rumen fatty acid metabolism and fatty acid composition in longissimus muscle and subcutaneous fat of lambs

To study the effect of feeding silages with different botanical composition, on rumen and lamb fat, 30 male lambs were assigned to five different silage groups for 11 weeks: botanically diverse silage (BDS); white clover silage (WCS); red clover silage (RCS), intensive English ryegrass silage (IRS) and crushed linseed and maize silage (MSL). Besides the silages, animals received organic wheat and barley and the MSL group additionally received bicarbonate (15 g/day). Silages were sampled when the bales were opened and analysed for fatty acid (FA) content and chemical composition. At slaughter, ruminal contents were sampled and 24 h after slaughter, longissimus muscle and subcutaneous (SC) fat were sampled. All samples were analysed for FA composition. The MSL group ingested the highest amount of FA (35.8 g/day v. 13.5, 19.4, 17.2 and 30.4 g/day for MSL v. BDS, WCS, RCS and IRS, respectively) and the sum of the major polyunsaturated FA, C18:2 n-6 and C18:3 n-3, was similar for groups BDS, WCS, RCS and MSL (61.3 g/100 g, 62.3 g/100 g, 62.3 g/100 g, 63.7 g/100 g of FA methylesters (FAME), respectively), while group IRS ingested higher proportions of these FA (74.5 g/100 g of FAME). Rumen data showed that animals fed BDS presented higher proportions of biohydrogenation intermediates, particularly C18:1 t11 and CLA c9t11, suggesting partial inhibition of rumen biohydrogenation. In the MSL group, the content of C18:3 n-3 in the rumen was highest, most probably due to reduced lipolysis and hence biohydrogenation through the combined effect of esterified C18:3 n-3 and seed protection. Additionally, C18:3 n-3 proportions were higher in rumen contents of RCS animals compared with WCS animals, which could be due to the activity of the polyphenol oxidase enzyme in the RC silages. Proportions of C18:3 n-3 were similar between treatments both for SC and intramuscular (IM) fat, whereas CLA c9t11 content was higher in the SC fat of BDS animals and lower in the IM fat of IRS animals compared with the other forage groups. No differences were found for C20:4 n-6, C20:5 n-3, C22:5 n-3 and C22:6 n-3 in the IM fat of the animals. Nevertheless, indices for desaturation and elongation activity in muscle of BDS animals suggest some stimulation of the first three steps of desaturation and elongation (Δ6-desaturase, elongase and Δ5-desaturase) of long-chain FA.     

Isolation and Characterization of a Δ5-Desaturase from Oblongichytrium sp.

We isolated a cDNA clone with homology to known desaturase genes from Oblongichytrium sp., recently classified as a new genus of thraustochytrids (Labyrinthulomycetes), and found that it encoded Δ5-desaturase by its heterologous expression in yeast. The enzyme had higher activity toward 20:4n-3 than 20:3n-6, indicating that this Δ5-desaturase can be used in the production of n-3 polyunsaturated fatty acids in transgenic organisms.     

Metabolism of unusual membrane phospholipids in the marine sponge Microciona prolifera

Sponges are unique in regard to membrane phospholipid composition. Features virtually without parallel in other organisms are the predominance of the C26-C30 polyenoic acids (demospongic acids) in the phosphatidylethanolamines (PE) and the attachment of identical acyl groups to the glycerol moiety. The biosynthesis and disposition of these unusual phospholipids were followed in the marine sponge Microciona prolifera where PE ( delta 5,9-26:2, delta 5,9-26:2) is a major molecular species. Incorporation experiments with radiolabeled fatty acids, bases, and intact phospholipids revealed the de novo biosynthesis of the two major phosphatides, phosphatidylethanolamines (PE) and phosphatidylcholines (PC), via the cytidine pathway as in higher animals, with ethanolamine selectively incorporated into PE( delta 5,9-26:2, delta 5,9-26:2). Methylation of PE and random acyl chain migration across different phospholipid classes were marginal, but the exchange of PC for PE, apparently mediated by the action of phospholipase, was indicated after uptake of the unnatural PC( delta 9-27:1, delta 9-26:1). The present study demonstrates in the most primitive multicellular animals a phospholipid metabolic pattern similar to that in higher organisms, with unique acyl and phosphoethanolamine transferases apparently involved in the biosynthesis of the (demospongic) di-C26-acyl-PE molecular species.     

First evidence of the presence of chitin in skeletons of marine sponges. Part II. Glass sponges (Hexactinellida: Porifera)

Sponges (Porifera) are presently gaining increased scientific attention because of their secondary metabolites and specific skeleton structures. In contrast to demosponges, whose skeletons are formed from biopolymer spongin, glass sponges (hexactinellids) possess silica-organic composites as the main natural material for their skeletal fibres. Chitin has a crystalline structure and it constitutes a network of organized fibres. This structure confers rigidity and resistance to organisms that contain it, including monocellular (yeast, amoeba, diatoms) and multicellular (higher fungi, arthropods, nematodes, molluscs) organisms. In contrast to different marine invertebrates whose exoskeletons are built of chitin, this polysaccharide has not been found previously as an endogenous biopolymer within glass sponges (Hexactinellida). We hypothesized that glass sponges, which are considered to be the most basal lineage of multicellular animals, must possess chitin. Here, we present a detailed study of the structural and physico-chemical properties of skeletal fragments of the glass sponge Farrea occa. We show that these fibres have a layered design with specific compositional variations in the chitin/silica composite. We applied an effective approach for the demineralization of glass sponge skeletal formations based on an etching procedure using alkali solutions. The results show unambiguously that alpha-chitin is an essential component of the skeletal structures of Hexactinellida. This is the first report of a silica-chitin's composite biomaterial found in nature. From this perspective, the view that silica-chitin scaffolds may be key templates for skeleton formation also in ancestral unicellular organisms, rather than silica-protein composites, emerges as a viable alternative hypothesis.     

Multiple defensive roles for triterpene glycosides from two Caribbean sponges

Despite their high nutritional value and a lack of physical defenses, most marine sponges appear to be minimally affected by predators, competitors, and fouling organisms, possibly due to sponge chemical defenses. In the last 15 years, several triterpene glycosides have been isolated from sponges, but their ecological or physiological roles are largely unknown. We tested triterpene glycosides from Erylus formosus and Ectyoplasia ferox, Caribbean sponges belonging to two different orders, in field and laboratory assays for effects on fish feeding, attachment by potential biofilm-forming bacteria, fouling by invertebrates and algae, and overgrowth by neighboring sponges. Formoside and other triterpene glycosides from Erylus formosus deterred predation, microbial attachment, and fouling by invertebrates and algae. Triterpene glycosides from Ectyoplasia ferox were found to be antipredatory and allelopathic. Thus, triterpene glycosides in these sponges appear to have multiple ecological functions. Tests with different triterpene glycosides at several concentrations indicated that small differences in molecular structure affect ecological activity. In order to establish whether triterpene glycosides could be involved in water-borne versus surface-mediated interactions, the presence of triterpene glycosides in the seawater surrounding live sponges was measured using two in situ sampling methods followed by HPLC and NMR spectral analysis. Water-borne triterpene glycosides were below detection limits for both species. However, top sponge layers and swabs of the surfaces of both sponges contained sufficiently high concentrations of triterpene glycosides to deter bacterial settlement and fouling of Erylus formosus surfaces and overgrowth of Ectyoplasia ferox by neighboring sponges. Enemies of these sponges appear to be deterred by surface contact of triterpene glycosides rather than by water-borne interactions. The dual strategy of employing one group of compounds for multiple purposes and minimizing the loss of compounds into seawater suggests that these organisms utilize chemical defenses with efficiency.     

Elevated cadmium accumulation in marine organisms from Terra Nova Bay (Antarctica)

 As a contribution towards identification of the principal environmental factors involved in cadmium accumulation in Antarctic marine organisms and the establishment of a baseline near the Italian Antarctic Station “Baia Terra Nova”, surface sediments, plankton and benthic organisms were studied in coastal waters of Terra Nova Bay (Ross Sea). The cadmium content of sediments was similar to that regarded as background in most marine coastal areas, whereas in surface water, phyto- and zooplankton it was similar to values measured in areas of enhanced upwelling. Algal and animal taxa dominating benthic associations had a higher cadmium content than related species from other seas. Very high concentrations of the metal were found in sponges (10–80 μg/g dw) and in the digestive gland of molluscs (up to 345 μg/g in Neobuccinum eatoni). The rapid regeneration of cadmium and its natural occurrence and bioavailability in highly productive coastal waters seem to be responsible for cadmium accumulation in the tissues of marine organisms near the “Baia Terra Nova” station. Received: 6 June 1995/Accepted: 23 November 1995     

Delta-8 desaturation activity varies among fatty acyl desaturases of teleost fish: high activity in delta-6 desaturases of marine species

The benefits of dietary fish and fish oil are derived from n-3 long-chain polyunsaturated fatty acids (LC-PUFA) that have beneficial effects in a range of human diseases and pathologies such as cardiovascular and other inflammatory disorders, neural development and neurological pathologies. The precursor of n-3 LC-PUFA, 18:3n-3 does not have the same beneficial effects prompting interest in the pathways of endogenous synthesis of LC-PUFA in vertebrates. The LC-PUFA biosynthesis pathway classically involves Δ6 and Δ5 fatty acyl desaturases (Fad), but it was recently shown that Δ6 Fad in mammals also displayed Δ8 activity demonstrating a possible alternative "Δ8-pathway" for the synthesis of LC-PUFA. Our primary hypothesis was that Δ8 desaturase activity would be a common feature of vertebrate Δ6 Fads, and so the aim of the present study was to determine the ability of teleostei Fads for Δ8 desaturation activity. To this end, cDNAs for Fads from a range of freshwater, diadromous and marine teleost fish species were assayed for Δ8 activity in the heterologous yeast expression system. In summary, the present study has demonstrated that Δ8 desaturation activity was also a characteristic of fish orthologs, although the activity varied notably between freshwater/diadromous and marine fish species, with the latter possessing Fads2-like proteins with Δ8 activity far higher than mammalian FADS2. The data showed that, generally, the fish Fad are technically υ-3 desaturases, with new double bonds introduced 3C beyond a pre-existing double bond. However, the ability of zebrafish and rabbitfish Fads, previously characterised as Δ6/Δ5 bifunctional desaturases, to introduce non-methylene interrupted double bonds in 20:3n-3 and 20:2n-6 suggested that a novel combination of regioselectivity modes operates within these enzymes.     

Combined measurements of egg fatty acids and stable isotopes as indicators of feeding ecology in lake-dwelling birds

1. We examined whether egg fatty acid (FA) profiles and stable isotopes (SIA) could be used in a comparative way to infer the diet of two aquatic bird species with contrasting feeding habits: a surface forager, the pheasant-tailed jacana ( Hydrophasianus chirurgus ), and a pursuit-diving forager, the little cormorant ( Phalacrocorax niger ), at Haleji Lake (Pakistan).
2. The species differed markedly in the overall percentage of saturated and polyunsaturated fatty acids. Moreover, among jacanas, two groups of birds had relatively high or low concentrations of long-chain essential PUFAs (such as 18:2 n-6 and 20:4 n-6), suggesting differing contributions from animal prey and plant material.
3. These trophic differences were corroborated by δ¹⁵N values which indicated both a higher trophic position in cormorants relative to jacanas, and differences in trophic position for the two groups of jacanas. In this latter species δ¹³C values in both groups also pointed to differing diets, involving mainly grazing plants or a contribution from animal resources (aquatic invertebrates or insects).
4. Both lower values of δ¹³C and higher percentages of 18:1 n-7 detected in little cormorants may indicate the influence of the anoxic water, typical of a freshwater system densely covered by macrophytes.
5. These results indicate how both biomarkers offer complementary information in studies of feeding ecology, refining interpretations of trophic pathways which are usually based on FA or SIA alone. Comparisons of FA profiles and SIA among species also proved to be a reliable approach for inferring the diet of species for which information is scarce or contradictory, as is the case for the pheasant-tailed jacana.