Fatty Acid Modification Research Articles

Transcriptional regulation of seed oil accumulation in Arabidopsis thaliana: role of transcription factors and chromatin remodelers

Triacylglycerols (TAGs) are derived from ester linkage of fatty acids (FAs) and glycerol and stored by plants in their seeds as carbon and energy resource to help seeds germinate and form self-sustainable seedlings. There are two main steps in TAG biosynthesis—biosynthesis of FAs in plastids and assembly of TAG in the endoplasmic reticulum. FA biosynthesis and TAG assembly is orchestrated by many transcription factors (TFs) which operate in gene regulatory networks by controlling the expression of some key genes coding for enzymes involved in the above processes. This review highlights the role of TFs involved in positive and negative transcriptional regulation of genes involved in accumulation of seed oil in Arabidopsis thaliana. Furthermore, epigenetic regulation of the expression of these key regulators of FA biosynthesis is elaborated. Also, the role of microRNAs in the regulation of target genes, including TFs, involved in FA biosynthesis, is discussed. Altogether, this review focuses on transcriptional regulation by TFs and chromatin remodelers, and post-transcriptional regulation of genes encoding for enzymes involved in FA biosynthesis and modifications by microRNAs. Manipulating the expression of these key factors can help increase seed oil content in oilseed crops.

N-terminal Myristoylation Enhanced the Antimicrobial Activity of Antimicrobial Peptide PMAP-36PW

Drug-resistant bacteria infections and drug residues have been increasing and causing antibiotic resistance and public health threats worldwide. Antimicrobial peptides (AMPs) are novel antimicrobial drugs with the potential to solve these problems. Here, a peptide based on our previously studied peptide PMAP-36PW was designed via N-terminal myristoylation and referred to as Myr-36PW. The fatty acid modification provided the as-prepared peptide with good stability and higher antimicrobial activity compared with PMAP-36PW in vitro. Moreover, Myr-36PW exhibited effective anti-biofilm activity against Gram-negative bacteria and may kill bacteria by improving the permeability of their membranes. In addition, the designed peptide Myr-36PW could inhibit the bacterial growth of Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa GIM 1.551 to target organs, decrease the inflammatory damage, show an impressive therapeutic effect on mouse pneumonia and peritonitis experiments, and promote abscess reduction and wound healing in infected mice. These results reveal that Myr-36PW is a promising antimicrobial agent against bacterial infections.

Rational design of a GLP-1/GIP/Gcg receptor triagonist to correct hyperglycemia, obesity and diabetic nephropathy in rodent animals

AimsTo design and screen a potent GLP-1/GIP/Gcg receptors triagonist with therapeutic potential in rodent animals with diabetes and obesity. Main methodsFirst, we obtained a 12-mer dual GIP/Gcg receptor agonist from a large combinatorial peptide library via high-throughput screening technique and then fused to the Exendin (9–39) to generate a potent GLP-1/GIP/Gcg triagonist. Further site fatty chain modification was performed to improve the druggability via enhancing in vivo stability and cyclic half-life. In vitro signaling and functional assays in cell lines expressing each receptor and in vivo efficacy evaluation in rodent model animals with hyperglycemia and obesity were all carefully performed. Key findingsWe screened and obtained a potent GLP-1/GIP/Gcg triagonist, termed XFL0, which promotes in vitro GLP-1, GIP, Gcg receptor activation comparable to native GLP-1, GIP and glucagon, respectively. Site-specific fatty acid modification significantly enhanced plasma stability of XFL0 and exhibited no obvious impact on receptor activation. The selected XFL0 conjugates termed XFL6, showed glucose-dependent insulin secretion and improved glucose tolerance by acting on all GLP-1, GIP and Gcg receptors in gene-deficient mice of which the effects were all significantly greater than any single receptor agonist. After chronic treatment in rodent animals with diabetes and obesity, XFL6 potently decreased body weight and food intake, ameliorated the hyperglycemia and hemoglobin A1c levels as well as the lipid metabolism and diabetic nephropathy related disorders. SignificanceXFL6, as a novel GLP-1/GIP/Gcg receptor triagonist, held potential to deliver outstanding improvement in correcting hyperglycemia, obesity and diabetic nephropathy.

Plastidic Δ6 Fatty-Acid Desaturases with Distinctive Substrate Specificity Regulate the Pool of C18-PUFAs in the Ancestral Picoalga Ostreococcus tauri.

Eukaryotic Δ6-desaturases are microsomal enzymes that balance the synthesis of ω-3 and ω-6 C18-polyunsaturated fatty acids (C18-PUFAs) according to their specificity. In several microalgae, including Ostreococcus tauri, plastidic C18-PUFAs are strictly regulated by environmental cues suggesting an autonomous control of Δ6-desaturation of plastidic PUFAs. Here, we identified two putative front-end Δ6/Δ8-desaturases from O tauri that, together with putative homologs, cluster apart from other characterized Δ6-desaturases. Both were plastid-located and unambiguously displayed a Δ6-desaturation activity when overexpressed in the heterologous hosts Nicotiana benthamiana and Synechocystis sp. PCC6803, as in the native host. Detailed lipid analyses of overexpressing lines unveiled distinctive ω-class specificities, and most interestingly pointed to the importance of the lipid head-group and the nonsubstrate acyl-chain for the desaturase efficiency. One desaturase displayed a broad specificity for plastidic lipids and a preference for ω-3 substrates, while the other was more selective for ω-6 substrates and for lipid classes including phosphatidylglycerol as well as the peculiar 16:4-galactolipid species occurring in the native host. Overexpression of both Δ6-desaturases in O tauri prevented the regulation of C18-PUFA under phosphate deprivation and triggered glycerolipid fatty-acid remodeling, without causing any obvious alteration in growth or photosynthesis. Tracking fatty-acid modifications in eukaryotic hosts further suggested the export of plastidic lipids to extraplastidic compartments.

Zebrafish (Danio rerio) physiological and behavioural responses to insect-based diets: a multidisciplinary approach

Black Soldier Fly (BSF) meal is considered as an alternative, emerging and sustainable ingredient for aquafeed production. However, results on fish physiological responses are still fragmentary and often controversial, while no studies are available on fish behavior in response to these new diets. The present work represents the first comprehensive multidisciplinary study aimed to investigate zebrafish physiological and behavioural responses to BSF-based diets. Five experimental diets characterized by increasing inclusion levels (0, 25, 50, 75 and 100% respect to fish meal) of full fat BSF prepupae meal were tested during a 2-months feeding trial. Prepupae were cultured on coffee silverskin growth substrate enriched with a 10% Schizochytrium sp. to improve insects’ fatty acids profile. The responses of zebrafish were assayed through biometric, histological, gas chromatographic, microbiological, spectroscopic, molecular and behavioural analyses. Results evidenced that BSF-based diets affected fish fatty acid composition, while behavioural tests did not show differences among groups. Specifically, a 50% BSF inclusion level diet represented the best compromise between ingredient sustainability and proper fish growth and welfare. Fish fed with higher BSF inclusions (75 and 100%) showed hepatic steatosis, microbiota modification, higher lipid content, fatty acid modification and higher expression of immune response markers.

Structural Insight into the Self-Assembly of a Pharmaceutically Optimized Insulin Analogue Obtained by Small-Angle X-ray Scattering.

B29Nε-lithocholyl-γ-l-βGlu-desB30 human insulin [NN344] belongs to a group of insulins with fatty acid or sterol modifications. These insulin analogues have been found to form subcutaneous depots upon injection and hereby have a protracted release profile in vivo. In the present study, B29Nε-lithocholyl-γ-l-Glu-desB30 human insulin was investigated using in-solution small-angle X-ray scattering (SAXS) at chemical conditions designed to mimic three stages during treatment in vivo: in-vial/pen, postinjection, and longer times after injection. We found that the specific insulin analogue formed a mixture of mono- and dihexamers under in-vial/pen conditions of low salt and stabilizing phenol. At postinjection, conditions mimicking a subcutaneous depot, B29Nε-lithocholyl-γ-l-Glu-desB30 human insulin, formed very long straight soluble hexamer-based rods stacked along the Zn(II)-axis. The self-assembly was triggered by an increase in salt concentration when going from vial to physiological conditions. Mimicking longer times after injection and the additional removal of phenol caused the length of the rods to decrease significantly. Finally, we found that the self-assembly could be controlled by varying the amount of modification at the interaction interface by making mixed hexamers of B29Nε-lithocholyl-γ-l-Glu-desB30 and desB30 human insulin. This opens extra possibilities for controlling the release profile of very-long-acting insulins.

A novel method for the modification of magnetite nanoparticles for the enhancement of its dispersibility in hydrophobic media

Different mixing methods were used for the fatty acid modification of magnetite nanoparticles. The magnetite nanoparticles were dissolved in ethanol with a previously dissolved oleic or stearic acid solution. The mix was then homogenized and washed to eliminate the unreacted fatty acid. The modified nanoparticles were FTIR analysis confirmed that the modified nanoparticles presented the characteristic groups of the fatty acids. The vibrating sample magnetometer analysis (VSM) showed a very slight decrease in the magnetic properties of the magnetite, from 88.7 to 53.7 emu/g, which obviously was due to the presence of the fatty acids. TGA showed that the fatty acid modification degree of the magnetite was less than that reported elsewhere, nonetheless, during the precipitation tests in toluene; it was observed that the modification was sufficient to maintain the stability of the dispersion without affecting the magnetic properties. Additionally, TEM micrographs showed that the modified magnetite nanoparticles formed a cluster, with the fatty acids surrounding the magnetite nanoparticles. Confirming that the fatty acid layer was deposited as monolayers on the magnetite particle surface of approximately 2 nm in thickness with OA and 2.3 nm with SA. It was observed that the use of the two fatty acids produced inter-digitation among the fatty acids. Finally, nanocomposites based on liquid silicone rubber (LSR) were prepared with modified and unmodified magnetite and their rheological and magnetic behavior was studied. This fatty acid modification of magnetite particles improved the dispersion and distribution of modified particles into the silicon matrix.

Unlocking the power of fatty acids as dietary tracers and metabolic signals in fishes and aquatic invertebrates.

Determining the transfer and transformation of organic matter in food webs is a fundamental challenge that has implications for sustainable management of ecosystems. Fatty acids (FA) offer a potential approach for resolving complex diet mixtures of organisms because they provide a suite of molecular tracers. Yet, uncertainties in the degree of their biochemical modification by consumers, due to selective retention or metabolism, have limited their application. Here, we consolidated 316 controlled feeding studies of aquatic ectotherms (fishes and invertebrates) involving 1404 species-diet combinations to assess the degree of trophic modification of FA in muscle tissue. We found a high degree of variability within and among taxa in the %FA in consumer muscle tissue versus %FA in diet regression equations. Most saturated FA had weak relationships with the diet (r2 < 0.30) and shallow slopes (m < 0.30), suggesting a lack of retention in muscle when fed in increasing amounts. Contrarily, several essential FA, including linoleic (18:2n-6) and α-linolenic acid (18:3n-3), exhibited significant relationships with the diet (m > 0.35, r2 > 0.50), suggesting supply limitations and selective retention in muscle by consumers. For all FA, relationships strengthened with increasing taxonomic specificity. We also demonstrated the utility of new correction equations by calculating the potential contributions of approximately 20 prey items to the diet of selected species of generalist fishes using a FA mixing model. Our analyses further reveal how a broad range of fishes and invertebrates convert or store these compounds in muscle tissue to meet physiological needs and point to their power in resolving complex diets in aquatic food webs. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Fatty acid metabolism and modifications in Chironomus riparius.

A priori knowledge of fatty acid modifications in consumers is essential for studies using fatty acids as biomarkers. We investigated fatty acid metabolism and possible modification pathways in benthic invertebrate Chironomus riparius larvae (Diptera). We conducted diet manipulation experiments using natural food sources (two chlorophyte algae, a diatom and a non-toxic cyanobacterium). We also did a diet-switch experiment on two different resources, fish food flakes TetraMin® and cyanobacterium Spirulina, to study fatty acid turnover in Chironomus. Results of the diet manipulation experiments indicate that Chironomus larvae have a strong tendency to biosynthesize 20:5n-3 and 20:4n-6 from precursor fatty acids, and that the dietary availability of polyunsaturated fatty acids (PUFA) does not control larval growth. Fatty acid modifications explain why low dietary availability of PUFA did not significantly limit growth. This has ecologically relevant implications on the role of benthic chironomids in conveying energy to upper trophic level consumers. A diet-switch experiment showed that the turnover rate of fatty acids in Chironomus is relatively fast--a few days. The compositional differences of algal diets were large enough to separate Chironomus larvae into distinct groups even if significant modification of PUFA was observed. In summary, fatty acids are excellent dietary biomarkers for Chironomus, if modifications of PUFA are considered, and will provide high-resolution data on resource use. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Dietary fat concentrations influence fatty acid assimilation patterns in Atlantic pollock (Pollachius virens).

A key aspect in the use of fatty acids (FA) to estimate predator diets using quantitative FA signature analysis (QFASA) is the ability to account for FA assimilation through the use of calibration coefficients (CC). Here, we tested the assumption that CC are independent of dietary fat concentrations by feeding Atlantic pollock (Pollachius virens) three formulated diets with very similar FA proportions but different fat concentrations (5-9% of diet) for 20 weeks. CC calculated using FA profiles of diet and triacylglycerols in pollock liver were significantly different for the three diets. To test the robustness of diet estimates to these differences, we used the CC set derived from feeding the diet with the lowest fat concentration, published prey FA profiles and realistic diet estimates of pollock to construct 'pseudo-predators'. Application of QFASA to each pseudo-predator using the three sets of CC and the same prey FA profiles resulted in diet estimate biases of twofold for major prey items and approximately fivefold for minor prey items. This work illustrates the importance of incorporating diets with fat concentrations that are similar to natural prey when conducting feeding experiments to calculate CC. This article is part of the theme 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Shifting systems: prerequisites for the application of quantitative fatty acid signature analysis in soil food webs

Quantitative fatty acid signature analysis (QFASA) is widely used to investigate trophic interactions in marine ecosystems, as nutritionally important ω3 long-chain polyunsaturated fatty acids at the food web base allow tracing of their trophic transfer in the food chain. By contrast, the basal resources in soil food webs comprise a wider array of trophic markers, including branched-chain, cyclopropane as well as several mono- and polyunsaturated fatty acids. These diverse markers allow distinguishing between the three dominant soil carbon and energy channels, the root, bacterial and fungal pathway. QFASA has not been applied yet to soil ecosystems owing to the lack of a priori data to fit the model. The present work investigates the transfer of absolute and relative trophic marker fatty acids into Collembola as dominant representatives of the soil mesofauna. Three different species were fed on a variety of single diets characteristic for the green and brown food chain. Calibration coefficients were calculated and diet estimation trials for mixed diet set-ups were performed, using a library comprising 50 different resources. However, estimation of Collembola diet was only partially successful, identifying the main components, but not the correct relative proportions. Adjustments by fat content or diet group exclusion did not improve the results. Nonetheless, this work provides, to our knowledge, a first comprehensive dataset to translate the application of QFASA from marine to soil ecosystems. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Fatty acid bioconversion in harpacticoid copepods in a changing environment: a transcriptomic approach

By 2100, global warming is predicted to significantly reduce the capacity of marine primary producers for long-chain polyunsaturated fatty acid (LC-PUFA) synthesis. Primary consumers such as harpacticoid copepods (Crustacea) might mitigate the resulting adverse effects on the food web by increased LC-PUFA bioconversion. Here, we present a high-quality de novo transcriptome assembly of the copepod Platychelipus littoralis, exposed to changes in both temperature (+3°C) and dietary LC-PUFA availability. Using this transcriptome, we detected multiple transcripts putatively coding for LC-PUFA-bioconverting front-end fatty acid (FA) desaturases and elongases, and performed phylogenetic analyses to identify their relationship with sequences of other (crustacean) taxa. While temperature affected the absolute FA concentrations in copepods, LC-PUFA levels remained unaltered even when copepods were fed an LC-PUFA-deficient diet. While this suggests plasticity of LC-PUFA bioconversion within P. littoralis, none of the putative front-end desaturase or elongase transcripts was differentially expressed under the applied treatments. Nevertheless, the transcriptome presented here provides a sound basis for future ecophysiological research on harpacticoid copepods. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Inter- and intraspecific differences in rotifer fatty acid composition during acclimation to low-quality food.

Biochemical food quality constraints affect the performance of consumers and mediate trait variation among and within consumer species. To assess inter- and intraspecific differences in fatty acid retention and conversion in freshwater rotifers, we provided four strains of two closely related rotifer species, Brachionus calyciflorus sensu stricto and Brachionus fernandoi, with food algae differing in their fatty acid composition. The rotifers grazed for 5 days on either Nannochloropsis limnetica or Monoraphidium minutum, two food algae with distinct polyunsaturated fatty acid (PUFA) profiles, before the diets were switched to PUFA-free Synechococcus elongatus, which was provided for three more days. We found between- and within-species differences in rotifer fatty acid compositions on the respective food sources and, in particular, highly specific acclimation reactions to the PUFA-free diet. The different reactions indicate inter- but also intraspecific differences in physiological traits, such as PUFA retention, allocation and bioconversion capacities, within the genus Brachionus that are most likely accompanied by differences in their nutritional demands. Our data suggest that biochemical food quality constraints act differently on traits of closely related species and of strains of a particular species and thus might be involved in shaping ecological interactions and evolutionary processes. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Ambient temperature and algal prey type affect essential fatty acid incorporation and trophic upgrading in a herbivorous marine copepod.

The essential fatty acids (EFA) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are critical nutrients for all organisms, and the temperature sensitivity of their trophic transfer in marine systems is of concern because of rising ocean temperatures. Laboratory-reared copepodites of the marine calanoid Calanus finmarchicus were used to test the effects of temperature (at 6°C, 12°C and increasing temperature stress) and prey type (the dinoflagellate Heterocapsa triquetra and the diatom Thalassiosira weissflogii) on the extent and efficiency of dietary EPA and DHA incorporation from phytoplankton to copepods in a set of feeding experiments using 13C labelling. Temperature was a significant determinant of C. finmarchicus copepodites' EFA incorporation and gross growth efficiency, defined as the fraction of ingested EFA retained in copepod tissue. Ingestion and incorporation of both EFA were higher at warmer temperature, except in the case of DHA in copepods feeding on diatoms. DHA-associated growth efficiency was higher at the higher temperature for copepodites consuming the dinoflagellate, but temperature-related variation in algal EFA content was also a predictive factor. Moreover, our results strongly suggest that copepodites are capable of synthesizing EPA when consuming an EPA-depleted diet. Our study implies that the copepod link of marine food webs is resilient in terms of EFA transfer when confronted with alterations of ambient temperature and prey type availability. Measurements presented here are critical for estimating how EFA transfer dynamics respond to intra- and interannual environmental variability. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Assimilation and turnover rates of lipid compounds in dominant Antarctic copepods fed with 13 C-enriched diatoms

The study revealed species- and stage-specific differences in lipid accumulation of the dominant Antarctic copepods, the primarily herbivorous Calanoides acutus (copepodite stage V (CV), females) and the more omnivorous Calanus propinquus (females) storing wax esters and triacylglycerols, respectively, which were collected in summer (end of December). Feeding carbon-labelled diatoms to these copepods, 13C elucidated assimilation and turnover rates of copepod total lipids as well as specific fatty acids and alcohols. The 13C incorporation was monitored by compound-specific stable isotope analysis (CSIA). CV stages of C. acutus exhibited an intense total lipid turnover and 55% of total lipids were labelled after 9 days of feeding. By contrast, total lipid assimilation of female C. acutus and C. propinquus was lower with 29% and 32%, respectively. The major dietary fatty acids 16:0, 16:1(n − 7) and 20:5(n − 3) had high turnover rates in all specimens. In C. acutus CV, the high rates of the de novo synthesized long-chain monounsaturated fatty acids and alcohols 20:1(n − 9) and 22:1(n − 11) indicate intense lipid deposition, whereas these rates were low in females. The differences in lipid assimilation and turnover clearly show that the copepod species exhibit a high variability and plasticity to adapt their lipid production to their various life phases.This article is part of the theme issue ‘The next horizons for lipids as ‘trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

The critical importance of experimentation in biomarker-based trophic ecology.

Fatty acids are commonly used as biomarkers for making inferences about trophic relationships in aquatic and soil food webs. However, researchers are often unaware of the physiological constraints within organisms on the trophic transfer and modification of dietary biomarkers in consumers. Fatty acids are bioactive molecules, which have diverse structures and functions that both complicate and enhance their value as trophic tracers. For instance, consumers may synthesize confounding non-dietary sourced markers from precursor molecules, and environmental conditions also affect fatty acid composition. There is a vital need for more research on the uptake and transfer of trophic biomarkers in individual organisms in order to advance the field and make meaningful use of these tools at the scale of populations or ecosystems. This special issue is focused on controlled feeding experiments on a diverse taxonomic breadth of model consumers from freshwater, marine and soil ecosystems with a goal of creating a more integrated understanding of the connection between consumer physiology and trophic ecology. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Stable isotopes of fatty acids: current and future perspectives for advancing trophic ecology.

To understand consumer dietary requirements and resource use across ecosystems, researchers have employed a variety of methods, including bulk stable isotope and fatty acid composition analyses. Compound-specific stable isotope analysis (CSIA) of fatty acids combines both of these tools into an even more powerful method with the capacity to broaden our understanding of food web ecology and nutritional dynamics. Here, we provide an overview of the potential that CSIA studies hold and their constraints. We first review the use of fatty acid CSIA in ecology at the natural abundance level as well as enriched physiological tracers, and highlight the unique insights that CSIA of fatty acids can provide. Next, we evaluate methodological best practices when generating and interpreting CSIA data. We then introduce three cutting-edge methods: hydrogen CSIA of fatty acids, and fatty acid isotopomer and isotopologue analyses, which are not yet widely used in ecological studies, but hold the potential to address some of the limitations of current techniques. Finally, we address future priorities in the field of CSIA including: generating more data across a wider range of taxa; lowering costs and increasing laboratory availability; working across disciplinary and methodological boundaries; and combining approaches to answer macroevolutionary questions. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Dynamics of diet-egg transfer of fatty acids in the teleost fish, red drum (Sciaenops ocellatus).

Eggs of marine organisms are increasingly being recognized as important components of marine food webs. The degree to which egg fatty acid profiles reflect maternal diet fatty acid profiles, and therefore the value of fatty acids in eggs as trophic biomarkers, depends on the species' reproductive strategy and the extent of modification of ingested fatty acids. We measured the dynamics of transfer of recently ingested fatty acids to spawned eggs in a batch-spawning teleost, red drum (Sciaenops ocellatus). Results of 21 diet-shift experiments, from which the fatty acid profiles of the diets and eggs were compared, showed that 15 of 27 fatty acids measured (one saturated, two monounsaturated and 12 polyunsaturated fatty acids) in eggs were correlated with their levels in the recent diet, and the rate of incorporation into eggs was proportional to the magnitude of the diet shift. Large shifts in diet might occur naturally during spawning migrations or when prey communities vary over time. Results of this study indicate that fatty acids in red drum eggs can be useful for studying adult diet and exploring trophic linkages in marine systems. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Juvenile Dungeness crabs (Metacarcinus magister) selectively integrate and modify the fatty acids of their experimental diets.

Dungeness crabs (Metacarcinus magister) are ecologically and economically important in the coastal Northeast Pacific, yet relatively little is currently known about their feeding behaviour in the wild or their natural diet. Trophic biomarkers, such as fatty acids (FA), can be used to reveal trophic interactions. We used two feeding experiments to assess differences in FA composition of juvenile crabs fed different known foods to evaluate how they modify and integrate dietary FA into their own tissues and determine whether crab FA reflect diet changes over a six-week period. These experimental results were then compared with the FA signatures of wild caught juvenile crab with undetermined diets. We found that juvenile Dungeness crabs fed different foods assimilated dietary FA into their tissues and were distinct in their FA signatures when analysed with multivariate statistics. Experimentally fed juvenile crabs contained greater proportions of the most abundant long-chain polyunsaturated fatty acids (LCPUFA, >C20) than their foods. Crabs fed foods lacking in LCPUFA, particularly DHA (22:6ω3, docosahexaenoic acid), did not survive or grew slower than crabs fed other foods. This suggests that LCPUFA are physiologically important for this species and indicates biosynthesis of these FA does not occur or is not sufficient to meet their needs.This article is part of the theme issue ‘The next horizons for lipids as ‘trophic biomarkers’: evidence and significance of consumer modification of dietary fatty acids’.

Interaction between ω6 and ω3 fatty acids of different chain lengths regulates Atlantic salmon hepatic gene expression and muscle fatty acid profiles.

Atlantic salmon smolts (approx. 20-months old) were fed experimental diets with different combinations of omega-6:omega-3 fatty acids (FAs) (high-ω6, high-ω3, or balanced) and eicosapentaenoic acid plus docosahexaenoic acid (EPA + DHA) levels (0.3, 1.0 or 1.4%) for 12 weeks. Muscle FA (% total FA) reflected dietary C18-polyunsaturated FA; however, muscle EPA per cent and content (mg g-1) were not different in salmon fed high-ω3 or balanced diets. Muscle DHA per cent was similar among treatments, while DHA content increased in fish fed 1.4% EPA + DHA, compared with those fed 0.3-1.0% EPA + DHA combined with high-ω6 FA. Muscle 20:3ω6 (DGLA) content was highest in those fed high-ω6 with 0.3% EPA + DHA. Quantitative polymerase chain reaction analyses on liver RNA showed that the monounsaturated FA synthesis-related gene, scdb, was upregulated in fish fed 1.0% EPA + DHA with high-ω6 compared to those fed 0.3% EPA + DHA. In high-ω3-fed salmon, liver elovl2 transcript levels were higher with 0.3% EPA + DHA than with 1.0% EPA + DHA. In high-ω6-fed fish, elovl2 did not vary with EPA + DHA levels, but it was positively correlated with muscle ARA, 22:4ω3 and DGLA. These results suggest dietary 18:3ω3 elongation contributed to maintaining muscle EPA + DHA levels despite a two- to threefold change in dietary proportions, while 18:2ω6 with 0.3% EPA + DHA increased muscle DGLA more than arachidonic acid (ARA). Positive correlations between hepatic elovl2 and fabp10a with muscle ω6:ω3 and EPA + DHA + ARA, respectively, were confirmed by reanalysing data from a previous salmon trial with lower variations in dietary EPA + DHA and ω6:ω3 ratios. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.