Diet composition and seasonal feeding patterns of a freshwater ringed seal (Pusa hispida saimensis)

The Saimaa ringed seal (Pusa hispida saimensis) is one of the few freshwater seal populations worldwide. The major conservation issue of this critically endangered population is bycatch mortality. We used digestive tract content and stable isotopes (δ¹³C and δ¹⁵N) to estimate the diet and seasonal feeding patterns for gaining better understanding of the seals feeding habits and potential conservation implications. The diet was similar across age groups. Altogether 15 fish species were identified and the most important were smelt (Osmerus eperlanus), ruff (Gymnocephalus cernuus), perch (Perca fluviatilis), vendace (Coregonus albula) and cyprinids. The high δ¹⁵N values suggested that the seals lose weight during winter and spring. Additionally the drop in δ¹⁵N values indicated that pups start to recover from postweaning stress and gain weight around the age of 6 mo. The isotope values differed regionally, which emphasizes that samples from consumers and prey should be collected from the same regions to improve interpretation of the stable isotopic results. Overall, diet composition suggests minimal to nonexistent competition with commercial or recreational fishing. However, observed weight loss of pups during summer may be related to higher risk of bycatch and this should be taken into account when planning temporal fishing closures.     

Temporal variation in the vertical stratification of blubber fatty acids alters diet predictions for lactating Weddell seals

Fatty acid signature analysis of blubber has been used to study the foraging ecology of some marine mammals. However, species-specific information on fatty acid (FA) deposition, distribution and mobilization is required to develop further the application of FA as trophic markers within the marine environment. Blubber samples were collected from adult female Weddell seals post-parturition and end of lactation, and were divided into inner and outer half sections. We determined the degree to which there was vertical stratification in FA composition, and how this changed over the lactation period. Inner and outer layers of post-parturition blubber cores separated into two distinct groups. Sixty-two per cent of the dissimilarity between the two layers was accounted for by a higher abundance of monounsaturated fatty acids (18:1ω9c and 16:1ω7c) in the outer blubber layer, and more saturated fatty acids (16:0 and 14:0) in the inner layer. By end of lactation, the FA composition of the inner layer was different to post-parturition samples, and 20:5ω3 had the highest fractional mobilization of all FA. In contrast, the proportion of FA in the outer layer did not change, and there was more variability in the fractional mobilization of FA indicating mobilization was not uniform across the blubber layer. Dietary predictions changed considerably when highly mobilized FA were removed from analyses, and predictions were more consistent with previous dietary studies. The lack of uniformity in FA mobilization adds problems to the future use of FASA in dietary predictions, highlighting the need for more detailed information on FA mobilization.     

Stratification and intra- and inter-specific differences in fatty acid composition of common dolphin (Delphinus sp.) blubber: implications for dietary analysis

Sixty-five fatty acids were quantified in the blubber of common dolphins (Delphinus delphis, D. capensis) incidentally caught off the coast of southern California. Dolphins were grouped by sex, reproductive status and species, and a blubber sample was collected at a mid-lateral site located caudal to the trailing edge of the dorsal fin. Samples were divided horizontally into inner, middle and outer layers and gradients in fatty acid content (mass percent) were observed across the depth of the blubber. Levels of monounsaturated fatty acids were greatest in the outer layer, whereas levels of saturated and polyunsaturated fatty acids were greatest in the inner layer. Degree of stratification was greatest in sexually mature dolphins. Blubber of sexually immature, but physically mature, male dolphins was also highly stratified, suggesting that this difference may be attributed to differences in diet. Classification and regression tree analysis resulted in the fewest misclassifications when dolphins were grouped by species, possibly indicating that these closely related animals forage on different prey species. Dietary-derived fatty acids were typically selected as splitting criteria in classification and regression tree analyses, suggesting that the observed differences in fatty acid composition between the various groups of dolphins may be attributed to differences in diet.     

Transfer of fatty acids from female seal blubber via milk to pup blubber

Fatty acids were determined by gas chromatography followed by principal component analysis (PCA) and soft independent modelling of class analogy (SIMCA) in the blubber of 18 female grey seals, Halichoerus grypus, in their milk and in the blubber of their 1-week-old nursing pups. Large individual differences were observed in both blubber and milk content of fatty acids. The fatty acid composition of the milk was systematically different from the composition in maternal blubber, with higher relative amounts of the saturated acids, the monounsaturated with 20 carbon atoms and the n3 polyunsaturated, except 18:3n3. The composition of the fatty acids in the blubber of the pups was different from that of the milk. The same fatty acids that were enriched in the milk were depleted in the blubber of the pups. Therefore the fatty acid composition in the blubber of the pups was similar to that in the adults, although not identical. The results from this investigation imply that the composition of the fatty acids in the blubber of female seals and in the blubber of their pups cannot be determined directly by analysis of fatty acid composition of milk. Accepted: 16 February 2000     

Variation in blubber fatty acid composition among marine mammals in the Canadian Arctic

The composition of predator adipose stores can provide important insights into foraging patterns and the ecological relationships among species. We determined the fatty acid (FA) composition of 843 blubber samples from 80 bearded seals (Erignathus barbatus), 33 harbor seals (Phoca vitulina), 239 harp seals (Pagophilus groenlandicus), 32 hooded seals (Cystophora cristata), 281 ringed seals (Phoca hispida), 53 walruses (Odobenus rosmarus rosmarus), 105 beluga whales (Delphinapterus leucas), and 20 narwhals (Monodon monoceros) across the Canadian Arctic to examine patterns of variability among and within species. FA signatures accurately distinguished phocid seals, walruses, and whales. Belugas and narwhals had the most similar FA signatures of any two species, suggesting substantial overlap in their diets, especially in the narwhal‐wintering area off eastern Baffin Island. Among phocid seals, harp and hooded seals had the most similar FA signatures. Bearded seals were most similar to walruses, which was consistent with the benthic feeding habits of both species. Within species, geographic differences in FA signatures were found over both large (>4,000 km) and small (<100 km) spatial scales. Overall, within‐species differences were smaller than among‐species differences. In general, FA signature patterns were consistent with previous studies of the ecology and diets of arctic marine mammals.     

Essential fatty acid profiles of maturation feeds used in freshwater ornamental fish culture

An investigation of live or freshly prepared feeds used as maturationdiets for freshwater ornamental fish was conducted to uncover similaritiesor differences in total and essential fatty acids. Analysis of these maturation feeds reveals that there are relatively low levels of totalfatty acids (0.81–8.96 mg/100 mg^–1 dry weight) and withthe exception of the beef heart diet all other feeds have undetectablelevels of docosahexaenoate (22:6n3). The beef heart diet was observed topossess 4.86 mg 100 mg^–1 dry weight of 22:6n3 most probablydue to the addition of skipjack tuna, Katsuwanus pelamis, roe. All otherfeeds examined were found to contain low to moderate levels ofeicosapentaenoate (20:5n3) 0.00–0.61 mg/100 mg^–1 dryweight. Surprisingly relatively high amounts of arachidonate (20:4n6)0.16–0.90 mg/100 mg^–1 dry weight were observed in all ofthe maturation diets and ranged between 3.79%–27.16% ona percent composition basis. The results obtained to date indicate a need toscrutinize the role of arachidonate in the maturation and spawningprocess.     

Variation in the fatty acid composition of blubber in Cape fur seals (<Emphasis Type="Italic">Arctocephalus pusillus pusillus</Emphasis>) and the implications for dietary interpretation

Analysis of the fatty acid (FA) composition of blubber is a valuable tool in interpreting the diet of marine mammals. This technique is based on the principle that particular FA present in prey can be incorporated largely untransformed into predator adipose tissue stores, thereby providing biochemical signatures with which to identify prey species. Several studies of phocid seals and cetaceans have documented vertical stratification in the FA composition of blubber such that inferences about diet may vary greatly depending on the layer of the blubber that is analysed. It is not known whether blubber in otariid seals (fur seals and sea lions) also displays vertical stratification in FA composition. Furthermore, it is not known whether the FA composition of blubber is uniform in these species. In the present study, the vertical and regional variation in FA composition of blubber was investigated in seven adult female Cape fur seals (Arctocephalus pusillus pusillus). The proportion of monounsaturated fatty acids (MUFA) was greater in the outer (43.6±1.3%) than inner portion (40.9±1.2%; t₂₀=5.59, P<0.001) whereas the proportions were greater in the inner than outer portions for saturated fatty acids (23.6±0.5% and 21.9±0.6%, respectively, t₂₀ = 5.31, P<0.001) and polyunsaturated fatty acids (PUFA, 35.5±0.7% and 34.5±0.7%, respectively, t₂₀ = 3.81, P < 0.001). There was an inverse relationship between MUFA and PUFA in the blubber, independent of sampling location. In addition, with the exception of the inner portion from non-lactating females, blubber from the mammary area had the highest proportions of 18:19c and total MUFA, followed by blubber from the rump and neck, suggesting that the deposition and mobilisation of blubber lipids may not be uniform around the body in otariid seals. These results support the need for blubber tissue to be sampled from the same site on animals, and to the full depth of the blubber layer, to minimise variation in FA profiles that could occur if different sites and depths were sampled. Such standardisation of sampling will further aid in interpreting diet in otariid seals using the FA Signature Analysis approach.     

Changes in fatty acid profiles of thermo-intolerant and thermo-tolerant marine diatoms during temperature stress

Fatty acid composition and degree of fatty acid saturation during temperature stress in thermo-intolerant (Phaeodactylum tricornutum) and thermo-tolerant (Chaetoceros muelleri) marine diatoms were investigated. A greater number of fatty acids were observed in C. muelleri than in P. tricornutum regardless of treatment. The major fatty acids detected were 14:0, 16:0, 16:1, 16:2, 16:3, 18:0, 18:1(n-9)c, 18:2(n-6) and 20:5(n-3) with additional fatty acids 18:1(n-9)t and 20:4(n-6) detected in C. muelleri. Short duration (2 h) temperature increase above optimal growth temperature had a greater effect on fatty acid composition in C. muelleri than in P. tricornutum and the degree of fatty acid saturation was affected more by temperature in C. muelleri than in P. tricornutum during both short and long duration (24 h) treatments. Total protein assay results suggest that P. tricornutum, but not C. muelleri, was undergoing stress under our growing conditions although lipids in both diatoms were affected by increased temperature. Immunodetection of proteins with anti-rubisco indicates that the rubisco large subunit was undergoing greater turnover in C. muelleri than in P. tricornutum. However, the integrity of rubisco as a suitable indicator of lipid status needs further study. This work supports the hypothesis that a particular temperature, and not treatment duration, has the greater effect on changes in fatty acid composition. Furthermore, changes in fatty acid composition and degree of fatty acid saturation occurred more quickly in the diatoms in response to increased temperature than previously observed in nutrient starvation studies. Since diatom lipids represent an important resource for growth and reproduction of marine animals, the rapid alteration of their lipid composition under temperatures normally encountered in marine environments warrants further study.     

Plasma non-esterified fatty acid profiles in wild Atlantic salmon during their freshwater migration and spawning

Total plasma non-esterified fatty acid (NEFA) levels increased significantly in adult Atlantic salmon during the first months of their upstream migration and spawning in the Exploits River, Newfoundland, Canada. The highest levels occurred in May and were 5467±270·43 nmol ml⁻¹ for females and 4617±334·70 nmol ml⁻¹ for males. Significantly higher levels were maintained by females compared with males for most of the upstream migration. Between August and October, total plasma NEFA levels declined by 61% in females but only 23% in males. The decline in plasma monounsaturated and polyunsaturated fatty acid levels accounted for 74% of the loss of NEFAs in females. Specific plasma NEFAs such as 16: 0 (palmitic), 16: 1 (palmitoleic), 18: 1n9 (oleic) and 20: 5n3 (eicosapentaenoic acid) differed significantly between males and females during migration and spawning. The mean gonadosomatic index ( I _G) values of females in May and just prior to spawning were 0·37±0·01 and 10·25±0·32, respectively. The rapid decline in the plasma NEFA content of females coincided with the largest increase in their I _G (1·85±0·02–10·25±0·32). Corresponding I _G values for males were 0·34±0·01 in May and 3·33±0·78 prior to spawning. Plasma NEFA levels of spent salmon did not differ between sexes and were significantly lower than those of salmon preparing to spawn.     

Identifying a reliable blubber measurement site to assess body condition in a marine mammal with topographically variable blubber,the Pacific walrus

Pacific walruses may be unable to meet caloric requirements in the changing Arctic ecosystem, which could affect body condition and have population‐level consequences. Body condition has historically been monitored by measuring blubber thickness over the xiphoid process (sternum). This may be an unreliable condition index because blubber at other sites along the body may be preferentially targeted to balance energetic demands. Animals in aquaria provided an opportunity for controlled study of how blubber topography is altered by caloric intake. Morphology, body mass, blubber thickness (21 sites), and caloric intake of five mature, nonpregnant, nonlactating female walruses were measured monthly (12 month minimum). Body condition (mass × standard length^?1) was described by a model that included caloric intake and a seasonal effect, and scaled positively with estimates of total blubber mass. Blubber thicknesses (1.91–10.69 cm) varied topographically and were similar to values reported for free‐ranging female walruses. Body condition was most closely related to blubber thickness measured dorsomedially in the region of the anterior insertion of the pectoral flippers (shoulders); sternum blubber thickness was a relatively poor indicator of condition. This study demonstrates the importance of validating condition metrics before using them to monitor free‐ranging populations.     

Using stable isotopes to understand changes in ringed seal foraging ecology as a response to a warming environment

Trends toward increased temperatures, reduced sea ice extent, and longer open water seasons have resulted in changing Arctic ecosystem dynamics. Expected changes include shifts in distribution and abundance of prey species for seabirds and marine mammals. Using stable isotope analysis, we studied spatial and interannual variation in ringed seal (Pusa hispida) feeding ecology in Hudson Bay in relation to environmental variables, between 2003 and 2010. Ringed seal muscle and hair samples collected from Arviat and Sanikiluaq, Nunavut, were analyzed for stable isotope ratios of nitrogen (δ¹⁵N) and carbon (δ¹³C). Seals from western Hudson Bay (Arviat) had higher δ¹⁵N and lower δ¹³C than seals from eastern Hudson Bay (Sanikiluaq), and stable isotope ratios varied interannually within each region. Peak δ¹⁵N occurred in years with spring air temperatures between approximately ?5°C and ?2°C. This temperature range was characteristic of warm years in western Hudson Bay and cool years in eastern Hudson Bay. We hypothesize that the high δ¹⁵N observed in ringed seals is indicative of greater importance of capelin (Mallotus villosus) in ringed seal diet. A comparison of ringed seal isotopic niche widths indicated greater dietary differences between western and eastern Hudson Bay with warming, suggesting a possible ecological divergence related to climate change.     

Dynamics of blood chylomicron fatty acids in a marine carnivore: implications for lipid metabolism and quantitative estimation of predator diets

Blubber fatty acid(s) (FA) signatures can provide accurate estimates of predator diets using quantitative FA signature analysis, provided that aspects of predator FA metabolism are taken into account. Because the intestinal absorption of dietary FA and their incorporation into chylomicrons (the primary transport lipoproteins for dietary FA in the blood) may influence the relationship between FA composition in the diet and adipose tissue, we investigated the metabolism of individual FA at these early stages of assimilation. We also investigated the capacity of chylomicron signatures to provide quantitative estimates of prey composition of an experimental meal. Six captive juvenile grey seals (Halichoerus grypus) were fed either 2.3 kg (n=3) or 4.6 kg (n=3) of Atlantic herring (Clupea harengus). Although chylomicron FA signatures resembled diet signatures at all samplings, absolute differences were smallest at 3-h post-feeding, when chylomicrons were likely largest and had the greatest ratio of triacylglycerol to phospholipid FA. Specific FA that differed significantly between diet and chylomicron signatures reflected either input from endogenous sources or loss through peroxisomal -oxidation. When these aspects of metabolism were accounted for, the quantitative predictions of diet composition generated using chylomicron signatures were extremely accurate, even when tested against 28 other prey items.     

Modifying fatty acid profiles through a new cytokinin‐based plastid transformation system

The widespread use of herbicides and antibiotics for selection of transgenic plants has not been very successful with regard to commercialization and public acceptance. Hence, alternative selection systems are required. In this study, we describe the use of ipt, the bacterial gene encoding the enzyme isopentenyl transferase from Agrobacterium tumefaciens, as a positive selectable marker for plastid transformation. A comparison between the traditional spectinomycin‐based aadA selection system and the ipt selection system demonstrated that selection of transplastomic plants on medium lacking cytokinin was as effective as selection on medium containing spectinomycin. Proof of principle was demonstrated by transformation of the kasIII gene encoding 3‐ketoacyl acyl carrier protein synthase III into tobacco plastids. Transplastomic tobacco plants were readily obtained using the ipt selection system, and were phenotypically normal despite over‐expression of isopentenyl transferase. Over‐expression of KASIII resulted in a significant increase in 16:0 fatty acid levels, and a significant decrease in the levels of 18:0 and 18:1 fatty acids. Our study demonstrates use of a novel positive plastid transformation system that may be used for selection of transplastomic plants without affecting the expression of transgenes within the integrated vector cassette or the resulting activity of the encoded protein. This system has the potential to be applied to monocots, which are typically not amenable to traditional antibiotic‐based selection systems, and may be used in combination with a negative selectable marker as part of a two‐step selection system to obtain homoplasmic plant lines.     

Importance of biological and abiotic factors for geochemical cycling in a freshwater eutrophic lake

Here we report the results of a comprehensive biogeochemical monitoring of Rostherne Mere in 1998, including changes in dissolved oxygen, organic carbon and nitrogen, nitrate/nitrite, ammonia, Al, Na, S, K, Mg, Ca, Si, Fe, Mn, orthophosphate, particulate N & P, suspended solids, temperature, pH, chlorophyll-a and zooplankton. The results demonstrated the major influence of primary producers on the overall geochemical cycling of N, P and Si, and suggested that the significance of zooplankton might have been previously underestimated. For major anions and cations, however, the influence of biota on lake water concentrations appeared to be negligible, reflecting the fact that these chemicals were present far in excess of plankton requirements. Thus changes in concentrations of Ca, K, Na, Mg and S were rather limited and must have reflected changes in hydrological and meteorological parameters. K, however, demonstrated a transitional pattern, reflecting some influence of biological uptake. During the stratification period, the slow processes of bacterial decomposition in the hypolimnion gradually released chemicals contained in the materials accumulated in the bottom layer, remarkably increasing the concentrations of dissolved compounds of those elements present in amounts comparable with the pool stored in the sedimenting detritus (e.g. orthophosphate P, ammonia N, Si and DOC). The decomposition also resulted in a drop in the redox potential, followed by partial denitrification and chemical release from the sediments. The hypolimnion of the Mere was confirmed to remain at the stage of Mn release, characterised by accumulation of DOC, orthophosphates, ammonia and initial stages of denitrification. High levels of P released from the sediments during the stratification period suggest that the lake’s recovery after sewage diversion might be further delayed.     

Analysis of interspecific variation in relative fatty acid composition: use of flow cytometry to estimate unsaturation index and relative polyunsaturated fatty acid content in microalgae

Relative polyunsaturated fatty acid content and unsaturation index are very important composition variables in the use of microalgae both for animal and human nutrition and biofuel production. A readily available technique to rapidly and inexpensively estimate relative fatty acid composition is very important for mass screening of new strains for the production of different types of oil. This study demonstrates the validity of Nile Red staining and flow cytometry for quick estimation of unsaturation index and relative fatty acid content in microalgae. Nile Red staining allows polar and neutral lipid contents to be estimated, and in this study a significant correlation was observed between polar/neutral ratio and fatty acid content in the species studied, corresponding to higher polyunsaturated fatty acid content in the polar lipid fraction of microalgae. This technique enables quick estimation of relative polyunsaturated fatty acid content and interspecific variation, as well as variations caused by culture conditions. In the species studied, most variability in fatty acid composition was due to variation in monounsaturated and polyunsaturated fatty acids, with less variation observed in saturated fatty acid content.     

A comparison of the major lipid classes and fatty acid composition of marine unicellular cyanobacteria with freshwater species

The fatty acid composition of two motile (strains WH 8113 and WH 8103) and one nonmotile (strain WH 7803) marine cyanobacteria has been determined and compared with two freshwater unicellular Synechocystis species (strain PCC 6308 and PCC 6803). The fatty acid composition of lipid extracts of isolated membranes from Synechocystis PCC 6803 was found to be identical to that of whole cells. All the marine strains contained myristic acid (14:0) as the major fatty acid, with only traces of polyunsaturated fatty acids. This composition is similar to Synechocystis PCC 6308. The major lipid classes of the nonmotile marine strain were identified as digalactosyl diacylglycerol, monogalactosyl diacylglycerol, phosphatidylglycerol, and sulfoquinovosyl diacylglycerol, identical to those found in other cyanobacteria.Abbreviations DGDG Digalactosyl diacylglycerol - MGDG Monogalactosyldiacylglycerol - PG Phosphatidylglycerol - SGDG sulfoquinovosyl diacylglycerol - gc gas chromatography - ms mass spectrometry     

Comparison of fatty acid profiles of spawning and non-spawning Pacific herring, Clupea harengus pallasi

Crude lipid and fatty acid composition from liver, intestine, roe, milt and flesh of spawning and non-spawning Pacific herring (Clupea harengus pallasi) were examined to determine the relative effects of spawning on the nutritional value of herring. Depletion of lipid due to spawning condition was significant (P < 0.01) in all organ tissues and flesh of spawning herring. The lipid content ranged from an average of 1.9 to 3.4% (wet weight basis) in different organ tissues of spawning herring, to 10.5 to 16% in non-spawning fish. The fatty acid profile exhibited many differences in the relative distribution of individual fatty acids among organ tissues and between the two fish groups. Oleic acid (C18:1n-9), a major monounsaturated fatty acid (MUFA) found in all tissue lipids, decreased significantly (P < 0.01) in spawning fish. The two monoenes, C20:1n-9 and C22:1n-11, occurred at high concentrations in the flesh but at only minor proportion in the digestive organs and gonads. Spawning herring also had significantly (P < 0.01) higher polyunsaturated fatty acids (PUFA) content in the organ tissues, particularly in the milt and ovary, with docosahexaenoic acid (C22:6n-3, DHA) having the greatest proportion. Among the n-6 fatty acids, only C18:2n-6 and C20:4n-6 occurred at notable amounts and were present in higher proportions in spawning fish. We concluded that although relatively higher n-3 fatty acid content was found in the organ lipids of spawning herring, they are not an energy-dense prey food source due to the fact that both flesh and gonads contain a very low amount of lipid.     

Oxidation of short-chain fatty acids by sulfate-reducing bacteria in freshwater and in marine sediments

Colony counts of acetate-, propionate- and l-lactate-oxidizing sulfate-reducing bacteria in marine sediments were made. The vertical distribution of these organisms were equal for the three types considered. The highest numbers were found just beneath the border of aerobic and anaerobic layers.Anaerobic mineralization of acetate, propionate and l-lactate was studied in the presence and in the absence of sulfate. In freshwater and in marine sediments, acetate and propionate were oxidized completely with concomitant reduction of sulfate. l-Lactate was always fermented. Lactate-oxidizing, sulfate-reducing bacteria, belonging to the species Desulfovibrio desulfuricans, and lactate-fermenting bacteria were found in approximately equal amounts in the sediments. Acetate-oxidizing, sulfate-reducing bacteria could only be isolated from marine sediments, they belonged to the genus Desulfobacter and oxidized only acetate and ethanol by sulfate reduction. Propionate-oxidizing, sulfate-reducing bacteria belonged to the genus Desulfobulbus. They were isolated from freshwater as well as from marine sediments and showed a relatively large range of usable substrates: hydrogen, formate, propionate, l-lactate and ethanol were oxidized with concomitant sulfate reduction. l-Lactate and pyruvate could be fermented by most of the isolated strains.     

A method for the direct evaluation of the fatty acid status in a drop of blood from a fingertip in humans: applicability to nutritional and epidemiological studies

Several studies have shown that the fatty acid composition of circulating lipids reflects dietary fat intake, in turn being related to health status. The fatty acid composition of plasma lipids is therefore an important parameter in studies on dietary interventions. The aim of our study was to develop a rapid and inexpensive method for the analysis of circulating fatty acids applicable to large population groups. Drops of blood collected from fingertips have been directly subjected to transmethylation for gas chromatography analysis. This new method, validated for reproducibility, has been compared with the conventional method, based on withdrawal of blood from the antecubital vein followed by lipid extraction, and identical data have been obtained with the two techniques. Observed and predicted differences between blood and plasma fatty acids are related to the contribution of circulating cell membranes in blood. Finally the application of the methods to samples from 100 healthy subjects and the assessed correlation between dietary habits and blood fatty acid profiles demonstrate the validity of the new method and its applicability to nutritional and epidemiological studies.     

The first total synthesis of the marine fatty acid (+/-)-2-methoxy-13-methyltetradecanoic acid: a cytotoxic fatty acid to leukemia cells

The recently discovered marine fatty acid (+/-)-2-methoxy-13-methyltetradecanoic acid was synthesized for the first time in six steps (26% overall yield) starting from commercially available methyl 12-methyltridecanoate. The synthetic approach provided enough material to corroborate the structure of the acid, which was recently identified in the sponge Amphimedon complanata from Aguadilla, Puerto Rico, and to test its cytotoxicity to three leukemia cell lines. The key step in the synthesis was the addition of trimethylsilyl cyanide to 12-methyltridecanal under triethylamine catalysis. Nuclear magnetic resonance data are provided for the first time for this methoxylated fatty acid and the synthetic approach utilized is of general applicability since it can be used in the synthesis of other methyl-branched 2-methoxylated fatty acids. We also report that the acid (+/-)-2-methoxy-13-methyltetradecanoic acid is cytotoxic to human chronic myelogenous leukemia K-562 (EC50=238 microM), histiocytic lymphoma U-937 (EC50=250 microM), and promielocytic leukemia HL-60 (EC50=476 microM) in RPMI 1640 medium.