Long-chain Polyunsaturated Fatty Acids Research Articles

The importance of essential fatty acids and their ratios in aquafeeds to enhance salmonid production, welfare, and human health

Long chain polyunsaturated fatty acids (LC-PUFA), namely those from omega-3 (n-3) and omega-6 (n-6) families, are paramount for both fish and human nutrition. Some of these biomolecules cannot be synthesized de novo and must be acquired through the diet, being termed dietary essential fatty acids (EFA). Fish requirements for EFA have traditionally been met through the incorporation of fish oil (FO) in the formulation of aquafeeds. However, with limited supply of FO the aquaculture industry is searching for additional sustainable sources of LC-PUFA. This has significantly shifted the type of ingredients used in aquafeed formulation, namely vegetable oils (VO) deficient in long-chain omega-3, often resulting in imbalanced levels and ratios of fatty acid classes. Such imbalances can negatively affect fish performance and welfare, as well as the levels of health promoting omega-3 LC-PUFA present in fish fillets. Given the relevance that salmonid aquaculture plays in global fish production (principally Atlantic salmon, Salmo salar), as well as its growing role as a source of dietary health promoting omega-3 LC-PUFA for humans, the present review summarizes the scientific knowledge available to date on the dietary requirements for LC-PUFA by salmonids and humans. We discuss the implications of using imbalanced aquafeed formulations upon fish performance and welfare, as well as the subsequent consequences for human nutrition, along with current efforts to replace FO by alternative ingredients such as algal oil (AO) that can safeguard high-quality salmonid products for human consumption.

Biotechnology of α-linolenic acid in oilseed rape (Brassica napus) using FAD2 and FAD3 from chia (Salvia hispanica)

α-Linolenic acid (ALA, 18:3Δ9,12,15) is an essential fatty acid for humans since it is the precursor for the biosynthesis of omega-3 long-chain polyunsaturated fatty acids (LC-PUFA). Modern people generally suffer from deficiency of ALA because most staple food oils are low or lack ALA content. Biotechnological enrichment of ALA in staple oil crops is a promising strategy. Chia (Salvia hispanica) has the highest ALA content in its seed oil among known oil crops. In this study, the FAD2 and FAD3 genes from chia were engineered into a staple oil crop, oilseed rape (Brassica napus), via Agrobaterium tumefaciens-mediated transformation of their LP4-2A fusion gene construct driven by the seed-specific promoter PNapA. In seeds of T0, T1, and T2 lines, the average ALA contents were 20.86, 23.54, and 24.92%, respectively, which were 2.21, 2.68, and 3.03 folds of the non-transformed controls (9.42, 8.78, and 8.22%), respectively. The highest seed ALA levels of T0, T1, and T2 plants were 38.41, 35.98, and 39.19% respectively, which were 4.10–4.77 folds of the respective controls. FA-pathway enzyme genes (BnACCD, BnFATA, BnSAD, BnSCD, BnDGAT1, BnDGAT2, and BnDGAT3) and positive regulatory genes (BnWRI1, BnLEC1, BnL1L, BnLEC2, BnABI3, BnbZIP67, and BnMYB96) were all significantly up-regulated. In contrast, BnTT1, BnTT2, BnTT8, BnTT16, BnTTG1, and BnTTG2, encoding negative oil accumulation regulators but positive secondary metabolism regulators, were all significantly down-regulated. This means the foreign ShFAD2-ShFAD3 fusion gene, directly and indirectly, remodeled both positive and negative loci of the whole FA-related network in transgenic B. napus seeds.

Polyunsaturated Fatty Acids: Conversion to Lipid Mediators, Roles in Inflammatory Diseases and Dietary Sources.

Polyunsaturated fatty acids (PUFAs) are important components of the diet of mammals. Their role was first established when the essential fatty acids (EFAs) linoleic acid and α-linolenic acid were discovered nearly a century ago. However, most of the biochemical and physiological actions of PUFAs rely on their conversion to 20C or 22C acids and subsequent metabolism to lipid mediators. As a generalisation, lipid mediators formed from n-6 PUFAs are pro-inflammatory while those from n-3 PUFAs are anti-inflammatory or neutral. Apart from the actions of the classic eicosanoids or docosanoids, many newly discovered compounds are described as Specialised Pro-resolving Mediators (SPMs) which have been proposed to have a role in resolving inflammatory conditions such as infections and preventing them from becoming chronic. In addition, a large group of molecules, termed isoprostanes, can be generated by free radical reactions and these too have powerful properties towards inflammation. The ultimate source of n-3 and n-6 PUFAs are photosynthetic organisms which contain Δ-12 and Δ-15 desaturases, which are almost exclusively absent from animals. Moreover, the EFAs consumed from plant food are in competition with each other for conversion to lipid mediators. Thus, the relative amounts of n-3 and n-6 PUFAs in the diet are important. Furthermore, the conversion of the EFAs to 20C and 22C PUFAs in mammals is rather poor. Thus, there has been much interest recently in the use of algae, many of which make substantial quantities of long-chain PUFAs or in manipulating oil crops to make such acids. This is especially important because fish oils, which are their main source in human diets, are becoming limited. In this review, the metabolic conversion of PUFAs into different lipid mediators is described. Then, the biological roles and molecular mechanisms of such mediators in inflammatory diseases are outlined. Finally, natural sources of PUFAs (including 20 or 22 carbon compounds) are detailed, as well as recent efforts to increase their production.

Preparation of glycerides enriched with n-3 long-chain polyunsaturated fatty acids by Thermomyces lanuginosus lipase-catalyzed hydrolysis of tuna oil

n-3 Long-chain polyunsaturated fatty acids (LC-PUFAs) in glyceride form have attracted much attention due to their nutritional value and high bioavailability. In this work, Thermomyces lanuginosus lipase (TL 100 L) was used to selectively hydrolyze tuna oil for preparing glycerides enriched with n-3 LC-PUFAs. Effects of reaction variables on the hydrolysis reaction were optimized by response surface methodology, and the optimal conditions were as follows: reaction temperature 54 °C, enzyme load 3%, reaction time 6 h, mass ratio of water to tuna oil 1.5:1. Under these conditions, the resulting degree of hydrolysis was 55.01%, and the total content of eicosapentaenoic acid and docosahexaenoic acid in the glyceride fraction was found to be 46.39%. The final product was mainly composed of diacylglycerols (53.93%) and triacylglycerols (34.58%) after removal of the free fatty acids. This paper provides a simple and suitable method for industrial production of glycerides enriched with n-3 LC-PUFAs.

A simple method to isolate fatty acids and fatty alcohols from wax esters in a wax-ester rich marine oil.

Calanus finmarchicus is one of the most important zooplankton species in the North Atlantic. The zooplankton is currently being harvested and industrially processed to a marine oil product for human consumption as a marine nutraceutical containing long-chain omega-3 polyunsaturated fatty acids. This oil is very rich in wax esters, a lipid class where fatty acids are esterified to long chain fatty alcohols. In this paper we describe a simple method to 1) isolate the wax esters from the other lipid classes present in the oil, 2) hydrolyze the wax esters, and 3) separate the fatty acids from the fatty alcohol, all by means of solid phase extraction. Starting with an average of 322 mg Calanus oil, we obtained 75 mg fatty alcohols and 63 mg fatty acids. Contrary to previously described techniques, our method neither oxidize the fatty alcohols to fatty acids, nor are the fatty acids methylated, allowing the native, unesterified fatty acids and fatty alcohols to be used for further studies, such as in cell culture experiments to study the metabolic effects of these specific lipid fractions rather than the intact oil or wax esters.

Effects of a novel medium-long-medium-type structured lipid synthesized using a two-step enzymatic method on lipid metabolism and obesity protection in experimental mice.

We synthesized a novel, structured lipid containing caprylic acid at its sn-1,3 position and docosahexaenoic acid at its sn-2 position (1,3C-2D-TAG) by using a two-step enzymatic method and then investigated the relationship between the types of fatty acids in the structured lipid and their physiological functions. Furthermore, we compared the effects of similarly structured lipids on postprandial lipid metabolism and obesity protection. The results revealed that the novel structured lipid synthesized using the enzymatic method not only exhibited better physical properties than traditional oils but also had high oxidation stability and crystallization degree. In terms of physiological function, the low-dose 1,3C-2D-TAG group exhibited higher cholesterol and triglyceride levels, lower kidney weight or body weight, and higher serum aspartate aminotransferase and blood urea nitrogen levels than control group, whereas the high-dose 1,3C-2D-TAG group exhibited higher triglyceride levels. Moreover, the medium-dose 1,3C-2D-TAG group had remarkably higher high-density lipoprotein cholesterol levels and lower low-density lipoprotein cholesterol levels than the high-lipid, control, and 1,2,3C-TAG groups; however, the cholesterol and triglyceride levels and kidney and liver functions did not differ significantly among these groups. The study results suggest that 1,3C-2D-TAG can not only facilitate the effective utilization of the energy supplied by medium-chain fatty acids but also help overcome difficulties in the absorption of long-chain polyunsaturated fatty acids.

Post-feeding Molecular Responses of Cobia (Rachycentron canadum): RNA-Sequencing as a Tool to Evaluate Postprandial Effects in Hepatic Lipid Metabolism.

We used transcriptome sequencing to investigate the hepatic postprandial responses of Rachycentron canadum (cobia), an important commercial fish species. In total, 150 cobia juveniles (50 per tank, triplicate) were fed ad libitum with a commercial diet for 7days, fasted for 24h, and fed for 10min. The liver was sampled 10min prior to feeding and 30min, 1, 2, 4, 8, 12, and 24h after the feeding event. Each sample was evaluated in terms of liver fatty acid profile and gene expression. Differential gene expressions were evaluated, focusing on fatty acid synthesis and oxidation pathways. In general, the liver fatty acid profile reflected diet composition. Docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and n-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) levels increased at 8 to 12h but decreased at 24h after the feeding event. A high number of differentially expressed genes (DEGs) were observed comparing fish that fasted for 8h with those fasted for 30min and 24h, while a reduced number of DEGs was observed comparing individuals who fasted for 30min compared with those who fasted for 24h. Similarly, the main differences in the expression of genes related to the fatty acid biosynthesis and oxidation pathways were noticed in individuals who fasted for 8h compared with those who fasted for 30min and 24h. The results suggested that the adequate time to sample the individuals ranged between 8 and 12h after the meal since, apparently, after 24h, differential gene expression was not necessarily influenced by food intake.

Plankton depletion by mussel grazing negatively impacts the fitness of lobster larvae

Bivalve aquaculture has rapidly expanded in shallow coastal habitats, leading to substantial changes in the natural environment. While the grazing pressure of mussels (Mytilus edulis) on phytoplankton and zooplankton has been well documented, cascading effects of plankton depletion on the development and survival of resident fish and invertebrate larvae remains largely unexplored. This study experimentally assessed the impact of control (no mussel grazing), low (20 mussels·m−3) and high (80 mussels·m−3) mussel densities on a model community composed of various phytoplankton and zooplankton and how changes in plankton characteristics influence the development and survival of lobster (Homarus americanus) larvae. Mussels fed primarily on nanophytoplankton, while the picophytoplankton experienced significantly less grazing. Zooplankton was readily ingested by mussels, showing a sharp decline under low mussel density, whereas it was eliminated from the plankton under high density of mussels. A 65% decrease of polyunsaturated fatty acids (∑ PUFA) was observed in the plankton under mussel filtration compared to control. Lobster larvae reared under control plankton conditions exhibited the fastest development, highest survival rate, carapace length, total lipid and ∑ PUFA contents, and TAG/ST (triacylglycerols/sterols) ratio, indicating the best nutritional condition. Larvae reared under low mussel density showed longer developmental time, lower survival rate and carapace length than that for controls. In addition, these larvae showed a significant reduction in total lipid, ∑ PUFA, and TAG/ST, which collectively suggests poor nutritional status due to limited dietary intake. Under high mussel density, larvae did not molt to stage II and died within 14 days, suggesting that the depleted plankton community was unable to sustain physiological requirements for survival and development of the larvae. Overall, mussel grazing altered the meroplankton community structure and its biochemical composition, ultimately impacting the development, survival, and quality of lobster larvae. These findings provide novel information on complex trophic interactions between bivalves and a meroplankton species and adds a new perspective to fuel discussions on energy and nutrient pathways, particularly long-chain PUFA, in ecosystem carrying capacity studies for shellfish aquaculture in shallow coastal habitats.

Effect of dietary krill oil supplementation on horse red blood cell membrane fatty acid composition and blood parameters.

Supplementation with marine-derived n-3 long-chain polyunsaturated fatty acids (LC PUFAs), eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) is linked to beneficial health effects in both humans and horses. Krill oil (KO), which is extracted from the Antarctic krill (Euphausia superba), is well documented as a safe and biologically available dietary supplement in humans and several animal species, but there is a lack of documentation regarding its effect as a dietary ingredient for horses. The objective of this study was to test whether KO as a dietary supplement had the ability to raise horse red blood cell (RBC) membrane EPA and DHA, expressed as the n-3 index. Five nonworking Norwegian cold-blooded trotter horse geldings (body weight [BW]: 567 ± 38 kg) were supplemented with KO (10 mL/100 kg BW) for 35 days in a longitudinal study. Blood samples were analysed for RBC membrane fatty acid (FA) profile, haematology and serum biochemistry every 7th day. KO was well accepted by all horses, and no adverse health effects were observed during the 35-day trial period. KO supplementation affected the RBC membrane FA profile by increasing the n-3 index from Day 0 to 35 (Day 0: 0.53% vs. Day 35: 4.05% of total RBC FAs). The observed increase in the sum of EPA and DHA (p < 0.001), total n-3 FAs (p < 0.001) and the reduction of n-6 FAs (p < 0.044) resulted in a lower n-6:n-3 ratio (p < 0.001) by Day 35 of KO supplementation. In conclusion, the RBC n-3 index was increased and the general n-6:n-3 ratio was decreased in horses receiving 35-day dietary KO supplementation.

Docosahexaenoic acid promotes M2 microglia phenotype via activating PPARγ-mediated ERK/AKT pathway against cerebral ischemia-reperfusion injury

In ischemia-reperfusion stroke, microglia play a dual role in brain injury as well as brain repair, and promoting their switch from a pro-inflammatory M1 phenotype to an anti-inflammatory M2 phenotype is considered to be a potential therapeutic strategy. Docosahexaenoic acid (DHA) is an essential long-chain omega-3 polyunsaturated fatty acid that exhibits potent anti-inflammatory properties in the acute phase of ischemic stroke, but its effect on microglia polarization is unknown. Thus, the objective of this study was to investigate the neuroprotective effects of DHA on rat brain following ischemia-reperfusion injury, and to investigate the mechanism by which DHA regulates microglia polarization. We administered DHA 5 mg/kg intraperitoneally daily for 3 d following a transient middle cerebral artery occlusion reperfusion model in rats. The protective effects of DHA on cerebral ischemia-reperfusion injury were detected by TTC staining, HE staining, Nissler staining, and TUNEL staining. Quantitative real-time PCR, immunofluorescence, western blot, and enzyme-linked immunosorbent assay were used to detect the expression of M1 and M2 microglia-associated markers and PPARγ-mediated ERK/AKT signaling pathway proteins. We found that DHA significantly improved brain injury by decreasing the expression of the M1 phenotypic marker (iNOS, CD16) and increasing the expression of the M2 phenotypic marker (Arg-1, CD206). DHA also increased the expression of peroxisome proliferator-activated receptor gamma (PPARγ) mRNA and protein, increased the expression of the pathway protein AKT, and decreased the expression of ERK1/2. In addition, DHA promoted the expression of anti-inflammatory factor IL-10 and decreased the expression of pro-inflammatory factors TNF-α and IL-1β. However, the PPARγ antagonist GW9662 greatly blocked these beneficial effects. These results suggest that DHA may activate PPARγ to inhibit ERK and activate AKT signaling pathways to regulate microglia polarization, thereby reducing neuroinflammation and promoting neurological recovery to alleviate cerebral ischemia-reperfusion injury.

Omega-3 versus Omega-6: Are We Underestimating the Ecological Significance of Arachidonic Acid in Aquatic Systems?

The long-chain polyunsaturated fatty acids (PUFA) eicosapentaenoic acid (EPA, ω-3, or n-3) and arachidonic acid (ARA, ω-6 or n-6) are known to have distinct physiological functions, yet can both support growth and reproduction of consumers, raising the question of whether EPA and ARA are ecologically substitutable dietary resources. We explored the relative importance of EPA and ARA for the growth and reproduction of the freshwater keystone herbivore Daphnia in a life-history experiment. Both PUFA were supplemented in a concentration-dependent manner to a PUFA-free diet, separately and in combination (50% EPA: 50% ARA mixture). The growth-response curves obtained with EPA, ARA, and the mixture were virtually congruent and the thresholds for PUFA limitation did not differ, indicating that EPA (n-3) and ARA (n-6) were substitutable dietary resources under the applied experimental conditions. The actual requirements for EPA and ARA might change with growth conditions, e.g., under the influence of parasites or pathogens. The higher retention of ARA in Daphnia suggests that EPA and ARA are subject to different turnover rates, which also implies different physiological functions. Studies on the ARA requirements of Daphnia could provide valuable information on the presumably underestimated ecological importance of ARA in freshwater food webs.

Diet effects on longevity, heat tolerance, lipid peroxidation and mitochondrial membrane potential in Daphnia.

The dietary supply of polyunsaturated fatty acids (PUFA) crucially affects animals' performance at different temperatures. However, the underlying physiological mechanisms are still insufficiently understood. Here, we analyzed lifespan and heat tolerance of four genotypes of Daphnia magna reared on either the green alga Scenedesmus obliquus that lacks long-chain (> C18) PUFA, or the heterokont alga Nannochloropsis limnetica that contains C20 PUFA, both either at saturating and near-starvation levels. A significant genotype-by-diet interaction in lifespan was observed at saturating diets. The C20 PUFA-rich diet eliminated differences in lifespan among genotypes on the PUFA-deficient diet. Corrected for body length, acute heat tolerance was higher at low than at high food concentration, at least in the older of the two age groups analyzed. Genotypes differed significantly in heat tolerance, but there were no genotype-by-diet interactions. As predicted, the C20 PUFA-rich diet resulted in higher lipid peroxidation (LPO) and a lower mitochondrial membrane potential (ΔΨm). LPO levels averaged across clones and rearing conditions were inversely related to acute heat tolerance. Yet, heat tolerance was higher on the PUFA-rich diet than on the PUFA-deficient diet, particularly in older Daphnia, indicating that the C20 PUFA-rich diet allowed Daphnia to compensate for higher LPO. In contrast, Daphnia with intermediate levels of ΔΨm showed the lowest heat tolerance. Neither LPO nor ΔΨm explained the diet effects on lifespan. We hypothesize that antioxidants present in the PUFA-rich diet may have enabled higher heat tolerance of Daphnia despite higher LPO, which may also explain the lifespan expansion of otherwise short-lived genotypes.

Peanut thresholds in peanut-allergic children are related to dietary composition.

There is no clear explanation for the large variation in threshold levels among peanut-allergic children. We hypothesized that diet composition can partly explain this variation in thresholds, as nutrients and foods influence the intestinal barrier function and microbiota. to explore the relationship between the threshold levels for peanut and nutritional intake and gut microbial composition in peanut-allergic children. In this explorative cross-sectional study the cumulative threshold levels for peanut were determined by oral food challenge tests. Data on nutrients and foods consumed were obtained from 3-day food diaries. Microbial composition of faeces and saliva were determined by molecular microbiota detection technique. Multivariable linear regression analysis and multiple logistic regression were used to explore the associations, adjusted for energy and senitization. Sixty-five children were included, of whom 32 (49%) (median age 50 months, IQR 28.0-96.5) had a positive oral food challenge. Significant positive associations were found between the intake of total carbohydrates, vitamin A and cumulative threshold levels for peanut, while significant negative associations were found for long-chain polyunsaturated fatty acids, linoleic acid and omega-6 fatty acids. No associations were found between threshold levels and microbial composition of faeces and saliva. However, a significant higher abundance of Proteobacteria and Bacteroidetes in saliva (p = 0.011 and 0.04, respectively) and of Proteobacteria in faeces (p = 0.003) were found in children with a positive peanut challenge compared to children with a negative peanut challenge. As a novel concept, this study showed that dietary composition is related to threshold levels for peanut.

Impact of Omega-3 fatty acid Supplementation on Global Cognitive Function among Healthy and Althahimer’s disease Elders: A Systematic Review and Meta-analysis (P4-6.007)

<h3>Objective:</h3> This study investigated the role of LCn-3PUFA in global cognitive function among healthy and Alzheimer’s disease (AD) elders considering their baseline cognitive performance. <h3>Background:</h3> Cognitive decline is a core feature of the aging brain. Supplementation with Omega-3 long-chain polyunsaturated fatty acids (LCn-3PUFA) is widely used to manage dementia among elders, however the conflict about its effectiveness. <h3>Design/Methods:</h3> Five electronic databases were searched (PubMed, Scopus, Web of Science, Cochrane library, and Embase) for healthy or AD elders above 60 years old receiving LCn-3PUFA as a capsule supplementation, only randomized clinical trials that assess the global cognitive function were included, we investigated the global cognitive function by the mini-mental state examination scale (MMSE). <h3>Results:</h3> A total of 7409 citations were identified of which 13 citations were included in the final synthesis. The pooled Cohen’s d of MMSE favored the omega arm over the control arm in global cognitive performance (Cohen’s d −0.19, 95% CI (−0.27 to −0.10), P&lt;0.001), however, the studies were not homogeneous, the subgroup analysis showed that only healthy elders receiving LCn-3PUFA had significant results while no significant change was found among AD patients (Cohen’s d −0.20, 95% CI (−0.29 to −0.11), P&lt;0.001), (Cohen’s d −0.14, 95% CI (−0.36 to 0.09), P=0.16) respectively. Another subgroup analysis based on the baseline cognitive performance revealed that only healthy elders with no cognitive impairment showed significant results that favored the omega group (Cohen’s d −0.23, 95% CI (−0.33 to −0.13), P&lt;0.001), however, no significant change were recorded in elders with cognitive impairment ranging from mild to moderate (Cohen’s d −0.08, 95% CI (−0.25 to 0.09), P=0.33). <h3>Conclusions:</h3> Healthy elders with normal cognitive performance showed a significant benefit from LCn-3PUFA in global cognitive function, however, no significant change was recorded among elders with mild to moderate cognitive impairment or among elders with AD. <b>Disclosure:</b> Mr. Hefnawy has nothing to disclose. Dr. Diab has nothing to disclose. Dr. Elsayed has nothing to disclose. Dr. Abdelaziz has nothing to disclose. Dr. Mouffokes has nothing to disclose. Dr. Elboraay has nothing to disclose. Mr. Najah has nothing to disclose. Mr. Aljabali has nothing to disclose. Mr. Abdou Negida has nothing to disclose.

Transcriptome Analysis Reveals an Eicosapentaenoic Acid Accumulation Mechanism in a Schizochytrium sp. Mutant.

Eicosapentaenoic acid (EPA) is an omega-3 long-chain polyunsaturated fatty acid (PUFA) essential for human health. Schizochytrium is a marine eukaryote that has been widely utilized for the synthesis of PUFAs. The current low potency and performance of EPA production by fermentation of Schizochytrium spp. limits its prospect in commercial production of EPA. Since the synthesis pathway of EPA in Schizochytrium spp. is still unclear, mutagenesis combined with efficient screening methods are still desirable. In this study, a novel screening strategy was developed based on a two-step progressive mutagenesis method based on atmospheric and room temperature plasma (ARTP) and diethyl sulfate (DES) after multiple stresses (sethoxydim, triclosan and 2,2'-bipyridine) compound screening. Finally, the mutant strain DBT-64 with increased lipid (1.57-fold, 31.71 g/L) and EPA (5.64-fold, 1.86 g/L) production was screened from wild-type (W) strains; the docosahexaenoic acid (DHA) content of mutant DBT-64 (M) was 11.41% lower than that of wild-type strains. Comparative transcriptomic analysis showed that the expression of genes related to the polyketide synthase, fatty acid prolongation, and triglyceride synthesis pathways was significantly upregulated in the mutant strain, while the expression of genes involved in the β-oxidation pathway and fatty acid degradation pathway was downregulated in favor of EPA biosynthesis in Schizochytrium. This study provides an effective strain improvement method to enhance EPA accumulation in Schizochytrium spp. IMPORTANCE Schizochytrium, a marine eukaryotic microorganism, has emerged as a candidate for the commercial production of PUFAs. EPA is an omega-3 PUFA with preventive and therapeutic effects against cardiovascular diseases, schizophrenia, and other disorders. Currently, the low potency and performance of EPA production by Schizochytrium spp. limits its commercialization. In this study, we performed two-step progressive mutagenesis based on ARTP and DES and screened multiple stresses (sethoxydim, triclosan, and 2,2'-bipyridine) to obtain the EPA-high-yielding Schizochytrium mutant. In addition, high expression of the polyketide synthase pathway, fatty acid elongation pathway, and triglyceride synthesis pathway in the mutants was confirmed by transcriptomic analysis. Therefore, the multistress screening platform established in this study is important for breeding EPA-producing Schizochytrium spp. and provides valuable information for regulating the proportion of EPA in microalgal lipids by means of genetic engineering.

Reproduction of Hatchery-Reared Pike-Perch (Sander lucioperca) Fed Diet with Low-Marine-Ingredients: Role of Dietary Fatty Acids

This research aimed to evaluate the reproductive potential of hatchery-reared (F1) pike-perch (Sander lucioperca) broodstock fed a commercial diet with low levels of long-chain polyunsaturated fatty acids (Lc-PUFA) and wild (F0) pike-perch broodstock fed forage fish. Reproductive parameters, including pseudogonadosomatic index (PGSI), egg size, latency time, hatching rate, embryo survival, and eggs’ fatty acid (FA) composition, as well as plasma sex hormone, glucose and immunoglobulin levels after hormone injection, were analyzed. The results showed low PGSI (10% in F1 vs. 14% in F0) and embryo survival (24% in F1 vs. 61% in F0) in F1 broodstock, but a satisfactory hatching rate (63% in F1 vs. 78% in F0) and larval size (4.6 mm in F1 vs. 4.7 mm in F0). A low arachidonic acid (ARA) percentage in F1 fish eggs (1.32%), along with increased immunoglobulin levels (17.31 g/L), suggests that immune system activation might have depleted the reserves of ARA in F1 fish, which is the key fatty acid for successful oocyte maturation. We assumed that the administration of more sustainable diets, based on terrestrial plant ingredients, is not inferior to higher-quality diets based on marine ingredients.

Compound‐specific stable isotopes resolve sources and fate of polyunsaturated fatty acids in biota of headwater streams

Abstract Organisms at the base of stream food webs are typically poor in long‐chain polyunsaturated fatty acids (LC‐PUFA), especially in docosahexaenoic acid (DHA), whereas consumers at higher trophic levels are often rich in LC‐PUFA. For example, fish tissues, especially the brain, are DHA‐rich. This obvious mismatch between consumer LC‐PUFA and their basal dietary supply may result from selective retention and/or endogenous conversion of dietary precursors to LC‐PUFA. To determine which is more likely, we investigated compound‐specific carbon stable isotopes in PUFA (δ13CPUFA) of potential basal resources (stream epilithon, leaf litter) and consumers (invertebrates, European bullhead, and two salmonid species and their brain, eye, liver, and muscle tissues). We predicted that consumer‐PUFA, depleted in 13C values relative to their dietary sources, would indicate internal de novo PUFA synthesis. Alternatively, higher consumer‐δ13CPUFA values would imply selective retention of aquatic (epilithon and conditioned leaves) rather than terrestrial resources or internal production, irrespective of trophic levels. Invertebrate grazers and predators resembled δ13C values of essential fatty acids (δ13CEFA) of benthic algae, while shredder‐δ13CEFA values reflected those of conditioned rather than fresh leaves. Lower eye‐δ13CEFA values of salmonids than in livers indicated high retention of dietary PUFA sources (invertebrates and epilithon). Stable isotope values of eicosapentaenoic acid suggest that all consumers retained algal EPA, while insectivorous fish produced DHA in their liver. There is no further evidence from carbon stable isotopes for local PUFA conversion within neural tissues. Our study demonstrates that δ13CPUFA can be used to track sources of these highly functional molecules in aquatic consumers and highlights the importance of algal derived‐PUFA for these consumers in oligotrophic headwater streams.

Enteral long-chain polyunsaturated fatty acids and necrotizing enterocolitis: A systematic review and meta-analysis

BackgroundPreterm infants are at risk of long-chain polyunsaturated fatty acid (LCPUFA) deficiency. Recent studies on high-dose DHA; n-3 LCPUFA in preterm infants suggested potential positive effects on cognitive outcomes but raised concerns about some increased neonatal morbidities. These studies and recent recommendations for DHA supplementation generated controversy owing to the lack of balance between DHA and arachidonic acid (ARA; n-6 LCPUFA). ObjectivesTo identify the effect of enteral supplementation of DHA, with and without ARA, on necrotizing enterocolitis (NEC) in very preterm infants. MethodsA systematic review of randomized and controlled trials compared enteral LCPUFAs with placebo or no supplementation in very preterm infants. We searched PubMed, Ovid-MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, and CINHAL databases from inception to July 2022. Data were extracted in duplicate using a structured proforma. A meta-analysis and metaregression with random-effects models were used. The interventions evaluated were DHA alone vs. that combined with ARA, source of DHA, dose, and supplement delivery methods. Methodological qualities and risk of bias were assessed using the Cochrane risk-of-bias tool. ResultsFifteen randomized clinical trials (RCTs) included 3963 very preterm infants with 217 cases of NEC. Supplementation with DHA alone increased NEC (2620 infants; RR: 1.56; 95% CI: 1.02, 2.39) with no evidence of heterogeneity (I2 = 0.0%, P = 0.46). Multiple metaregression revealed significant reduction in NEC when ARA was supplemented with DHA (aRR 0.42; 95% CI: 0.21, 0.88). The source of DHA, dose, and feeding type revealed no associations with NEC. Two RCTs supplemented high-dose DHA to lactating mothers. There was a significant increase in risk of NEC with this approach (1148 infants; RR: 1.92; 95% CI: 1.02, 3.61) with no evidence of heterogeneity (I2 = 0.0, P = 0.81). ConclusionsSupplementation with DHA alone may increase risk of NEC. Concurrent supplementation with ARA needs to be considered when adding DHA to preterm infants’ diet.

Determination of dietary essential fatty acids in a deep-sea fish, the splendid alfonsino Beryx splendens: functional characterization of enzymes involved in long-chain polyunsaturated fatty acid biosynthesis

The splendid alfonsino Beryx splendens is a commercially important deep-sea fish in East Asian countries. Because the wild stock of this species has been declining, there is an urgent need to develop aquaculture systems. In the present study, we investigated the long-chain polyunsaturated fatty acid (LC-PUFA) requirements of B. splendens, which are known as essential dietary components in many carnivorous marine fish species. The fatty acid profiles of the muscles, liver, and stomach contents of B. splendens suggested that it acquires substantial levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from its natural diet. The functional characterization of a fatty acid desaturase (Fads2) and three elongases (Elovl5, Elovl4a, and Elovl4b) from B. splendens confirmed their enzymatic capabilities in LC-PUFA biosynthesis. Fads2 showed Δ6 and Δ8 bifunctional desaturase activities. Elovl5 showed preferential elongase activities toward C18 and C20 PUFA substrates, whereas Elovl4a and Elovl4b showed activities toward various C18-22 substrates. Given that Fads2 showed no Δ5 desaturase activity and no other fads-like sequence was found in the B. splendens genome, EPA and arachidonic acid cannot be synthesized from C18 precursors; hence, they can be categorized as dietary essential fatty acids in B. splendens. EPA can be converted into DHA in B. splendens via the so-called Sprecher pathway. However, given that fads2 is only expressed in the brain, it is unlikely that the capacity of B. splendens to biosynthesize DHA from EPA can fulfill its physiological requirements. These results will be useful to researchers developing B. splendens aquaculture methods.

Omega-3 Fatty Acids during Pregnancy in Indigenous Australian Women of the Gomeroi Gaaynggal Cohort.

Higher dietary intakes of Omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFAs) have been linked to lower rates of preterm birth and preeclampsia. The aim of this analysis was to describe dietary intake and fractions of red blood cell (RBC) membrane LC-PUFAs during pregnancy in a cohort of Indigenous Australian women. Maternal dietary intake was assessed using two validated dietary assessment tools and quantified using the AUSNUT (Australian Food and Nutrient) 2011-2013 database. Analysis from a 3-month food frequency questionnaire indicated that 83% of this cohort met national n-3 LC-PUFA recommendations, with 59% meeting alpha-linolenic acid (ALA) recommendations. No nutritional supplements used by the women contained n-3 LC-PUFAs. Over 90% of women had no detectable level of ALA in their RBC membranes, and the median Omega-3 Index was 5.5%. This analysis appears to illustrate a decline in concentrations of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) across gestation in women who had preterm birth. However, there was no visible trend in LC-PUFA fractions in women who experienced hypertension during pregnancy. Further research is needed to better understand the link between dietary intake of n-3 LC-PUFA-rich foods and the role of fatty acids in preterm birth and preeclampsia.