C20 Polyunsaturated Fatty Acids Research Articles

Nitrogen and fatty acid rumen metabolism in cattle offered high or low polyphenol oxidase red clover silage

Polyphenol oxidase (PPO) in red clover (RC) has been shown to reduce both lipolysis and proteolysis in silo and implicated (in vitro) in the rumen. However, all in vivo comparisons have compared RC with other forages, typically with lower levels of PPO, which brings in other confounding factors as to the cause for the greater protection of dietary nitrogen (N) and C18 polyunsaturated fatty acids (PUFA) on RC silage. This study compared two RC silages which when ensiled had contrasting PPO activities (RC+ and RC−) against a control of perennial ryegrass silage (PRG) to ascertain the effect of PPO activity on dietary N digestibility and PUFA biohydrogenation. Two studies were performed the first to investigate rumen and duodenal flow with six Hereford×Friesian steers, prepared with rumen and duodenal cannulae, and the second investigating whole tract N balance using six Holstein-Friesian non-lactating dairy cows. All diets were offered at a restricted level based on animal live weight with each experiment consisting of two 3×3 Latin squares using big bale silages ensiled in 2010 and 2011, respectively. For the first experiment digesta flow at the duodenum was estimated using a dual-phase marker system with ytterbium acetate and chromium ethylenediaminetetraacetic acid as particulate and liquid phase markers, respectively. Total N intake was higher on the RC silages in both experiments and higher on RC− than RC+. Rumen ammonia-N reflected intake with ammonia-N per unit of N intake lower on RC+ than RC−. Microbial N duodenal flow was comparable across all silage diets with non-microbial N higher on RC than the PRG with no difference between RC+ and RC−, even when reported on a N intake basis. C18 PUFA biohydrogenation was lower on RC silage diets than PRG but with no difference between RC+ and RC−. The N balance trial showed a greater retention of N on RC+ over RC−; however, this response is likely related to the difference in N intake over any PPO driven protection. The lack of difference between RC silages, despite contrasting levels of PPO, may reflect a similar level of protein-bound-phenol complexing determined in each RC silage. Previously this complexing has been associated with PPOs protection mechanism; however, this study has shown that protection is not related to total PPO activity.

One-pot Synthesis and Static Bulk Foam Studies of C18-conjugated Zwitterionic Surfactant

This study reports a one-pot-synthesis of C18-conjugated zwitterionic surfactant, linolyel dimethyl amidopropyl betaine (Lino-DAB) for application in oil and gas industry. The synthesis proceeded with the direct thermo-condensation of N,N-dimethyl-1,3-propandiamine with C18-polyunsaturated omega-6 fatty acid (linoleic acid) and subsequent treatment with sodium chloroacetate. The structure of the (Lino-DAB) products (19 g) was identified by 1H-NMR and FTIR experiments and by comparing with the other long chain zwitterion surfactants from the literature. The static bulk foam tests in the absence and presence of crude oil under range of reservoir conditions (0 - 3.5 % salinity and 25 – 90 °C temperature) were conducted, the results indicated that the high salinity and presence of crude oil were detrimental to the foamability of the Lino-DAB (0.2 %) at ambient condition (25 °C), but the foamability were significantly improved at the higher temperature (60 – 90 °C). Similarly, the Lino-DAB exhibits viscoelastic properties in brine solutions and has the ability to retain its viscosity by 10% at 90°C. Thus, this study demonstrates that Lino-DAB surfactant could have a potential application to enhance oil recovery particularly in foam assisted flooding.

A complete enzymatic capacity for long-chain polyunsaturated fatty acid biosynthesis is present in the Amazonian teleost tambaqui, Colossoma macropomum

In vertebrates, the essential fatty acids (FA) that satisfy the dietary requirements for a given species depend upon its desaturation and elongation capabilities to convert the C18 polyunsaturated fatty acids (PUFA), namely linoleic acid (LA, 18:2n-6) and α-linolenic acid (ALA, 18:3n-3), into the biologically active long-chain (C20-24) polyunsaturated fatty acids (LC-PUFA), including arachidonic acid (ARA, 20:4n-6), eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3). Recent studies have established that tambaqui (Colossoma macropomum), an important aquaculture-produced species in Brazil, is a herbivorous fish that can fulfil its essential FA requirements with dietary provision C18 PUFA LA and ALA, although the molecular mechanisms underpinning such ability remained unclear. The present study aimed at cloning and functionally characterizing genes encoding key desaturase and elongase enzymes, namely fads2, elovl5 and elovl2, involved in the LC-PUFA biosynthetic pathways in tambaqui. First, a fads2-like desaturase was isolated from tambaqui. When expressed in yeast, the tambaqui Fads2 showed Δ6, Δ5 and Δ8 desaturase capacities within the same enzyme, enabling all desaturation reactions required for ARA, EPA and DHA biosynthesis. Moreover, tambaqui possesses two elongases that are bona fide orthologs of elovl5 and elovl2. Their functional characterization confirmed that they can operate towards a variety of PUFA substrates with chain lengths ranging from 18 to 22 carbons. Overall our results provide compelling evidence that demonstrates that all the desaturase and elongase activities required to convert LA and ALA into ARA, EPA and DHA are present in tambaqui within the three genes studied herein, i.e. fads2, elovl5 and elovl2.

The Transcriptional Factor PPARαb Positively Regulates Elovl5 Elongase in Golden Pompano Trachinotus ovatus (Linnaeus 1758).

The nuclear peroxisome proliferator-activated receptors (PPARs) regulate the transcription of elongases of very long-chain fatty acids (Elovl), which are involved in polyunsaturated fatty acid (PUFA) biosynthesis in mammals. In the present study, we first characterized the function of Elovl5 elongase in Trachinotus ovatus. The functional study showed that ToElovl5 displayed high elongation activity toward C18 and C20 PUFA. To investigate whether PPARαb was a regulator of Elovl5, we also reported the sequence of T. ovatus PPARαb (ToPPARαb). The open reading frame (ORF) sequence encoded 469 amino acids possessing four typical characteristic domains, including an N-terminal hypervariable region, a DNA-binding domain (DBD), a flexible hinge domain and a ligand-binding domain (LBD). Thirdly, promoter activity experiments showed that the region from PGL3-basic-Elovl5-5 (-146 bp to +459 bp) was defined as the core promoter by progressive deletion mutation of Elovl5. Moreover, PPARαb overexpression led to a clear time-dependent enhancement of ToElovl5 promoter expression in HEK 293T cells. Fourth, the agonist of PPARαb prominently increased PPARαb and Elovl5 expression, while PPARαb depletion by RNAi or an inhibitor was correlated with a significant reduction of Elovl5 transcription in T. ovatus caudal fin cells (TOCF). In conclusion, the present study provides the first evidence of the positive regulation of Elovl5 transcription by PPARαb and contributes to a better understanding of the transcriptional mechanism of PPARαb in fish.

An Insect Prostaglandin E2 Synthase Acts in Immunity and Reproduction.

Eicosanoids, oxygenated metabolites of C20 polyunsaturated fatty acids (PUFAs), mediate fundamental physiological processes, including immune reactions and reproduction, in insects. Prostaglandins (PGs) make up one group of eicosanoids, of which PGE2 is a relatively well-known mediator in various insect taxa. While PG biosynthesis has been reported, the specific biosynthetic pathway for PGE2 is not known in insects. Here, we posed the hypothesis that Se-mPGES2 mediates biosynthesis of physiologically active PGE2 through its cognate protein. To test this hypothesis, we interrogated a transcriptome of the lepidopteran insect, Spodoptera exigua, to identify a candidate PGE2 synthase (Se-mPGES2) and analyzed its sequence and expression. Its predicted amino acid sequence contains a consensus thioredoxin homology sequence (Cys-x-x-Cys) responsible for catalytic activity along with an N-terminal membrane-associated hydrophobic domain and C-terminal cytosolic domain. It also shares sequence homology (36.5%) and shares almost overlapping three dimensional structures with a membrane-bound human PGES2 (mPGES2). Se-mPGES2 was expressed in all developmental stages with high peaks during the late larval instar and adult stages. Immune challenge significantly up-regulated its expression levels in hemocytes and fat body. Injecting double-stranded RNA (dsRNA) specific to Se-mPGES2 significantly impaired two cellular immune responses, hemocyte-spreading behavior and nodule formation following bacterial challenge. Humoral immunity was also significantly suppressed, registered as reduced phenoloxidase activity and antimicrobial peptide expression levels. The suppressed immune responses were reversed following PGE2, but not arachidonic acid (AA), treatments. RNAi treatments also reduced the egg-laying behavior of females. Control females mated with the RNAi-treated males led to substantially reduced egg-laying behavior, which was also reversed following PGE2 injections into females. These results strongly bolster our hypothesis that Se-mPGES2 acts in the biosynthesis of PGE2, a crucial biochemical signal mediating immune and reproductive physiology of S. exigua.

Polyunsaturated C18 fatty acids derivatized with Gly and Ile as an additional tool for studies of the catalytic evolution of fungal 8- and 9-dioxygenases

The fungal linoleate diol synthase (LDS) family contains over twenty characterized 8-, 9-, and 10-dioxygenases (DOX), usually fused to catalytically competent cytochromes P450. Crystal structures are not available, but indirect evidence suggests that linoleic acid enters the active site of 8R-DOX-LDS headfirst and enters 9S-DOX-allene oxide synthase (AOS) with the ω-end (tail) first. Fatty acids derivatized with amino acids can conceivably be used to study oxidation in tail first position by enzymes, which bind natural fatty acids headfirst. The results might reveal catalytic similarities of homologous enzymes. 8R-DOX-5,8-LDS oxidize 18:2n-6-Ile and 18:2n-6-Gly in tail first position to 9S-hydroperoxy metabolites, albeit with less position and stereo specificity than 9S-DOX-AOS. The oxygenation mechanism of 9S-DOX-AOS with antarafacial hydrogen abstraction at C-11 and oxygen insertion at C-9 was also retained. Two homologues, 8R-DOX-7,8-LDS and 8R-DOX-AOS, oxidized 18:2n-6-Ile and 18:2n-6-Gly at C-9, suggesting a conserved feature of 8R-DOX domains. 9R-DOX-AOS, with 54% sequence identity to 9S-DOX-AOS, did not oxidize the derivatized C18 fatty acids. 9Z,12Z-16:2, two carbon shorter than 18:n-6 from the ω-end, was rapidly metabolized to an α-ketol, but 7Z,10Z-16:2 was not a substrate. An unsaturated carbon chain from C-1 to C-8 was apparently more important than the configuration at the ω-end. 8R-DOX-LDS and 9R-DOX-AOS may thus bind 18:2n-6 in the same orientation. The oxidation of 18:2n-6 in straight or reverse head-to-tail positions illustrates evolutionary traits between 8- and 9-DOX domains. Fatty acids derivatized with amino acids provide a complementary tool for the analysis of evolution of enzymes.

Carbon cycling in the Sargasso Sea water column: Insights from lipid biomarkers in suspended particles

This study compares lipid biomarker and bulk constituents (organic carbon and δ13C, nitrogen and carbonate) in particles (30–4400 m depth) collected at the Oceanic Flux Program site in the northern Sargasso Sea off Bermuda during three periods of contrasting oceanographic conditions: following the spring bloom (April 2015), during the low productivity period (November 2015), and two weeks after the passage of the Category 3 Hurricane Nicole (October 2016). Lipids biomarkers were used to evaluate the contributions of phytoplankton, zooplankton and bacterial sources to the particulate organic carbon (POC), diagenetic transformations of suspended POC throughout the water column and within the nepheloid layer, and seasonal and non-seasonal temporal variability, including the impact of an extreme weather event.Depth profiles of lipid concentration and molecular composition showed seasonal and non-seasonal variability in particle composition throughout the water column. Higher lipid concentrations in April versus November 2015 reflected differences in plankton productivity, while relative abundances in diagnostic phytosterols, C18 polyunsaturated fatty acids and alkenones reflected differences in phytoplankton community structure. A rapid decrease in lipid biomarker concentrations below the euphotic zone was accompanied by a marked transition in lipid composition, e.g. increases in the relative percentages of cholesterol, C16-C18 fatty alcohols, odd- and branched- chain fatty acids. This reveals the intense remineralization of algal-derived carbon and fresh inputs of organic materials from zooplankton and microbial production within the upper mesopelagic zone. Particulate lipids within the bathypelagic zone reveal a close connectivity of deep particulate carbon composition with upper ocean properties, while compositional heterogeneity with depth reflects continuous particle turnover and de novo particle production throughout the deep water column. Additionally, depth trends in the within-class composition of fatty alcohols and bacterial fatty acids were observed, suggesting depth zonation of zooplankton and microbial community structure.Hurricane Nicole (October 2016) strongly impacted upper ocean physics and nutrient supply, resulting in a transient phytoplankton bloom/flux event that increased concentrations of fresh particulate phytodetritus (e.g., C18 polyunsaturated fatty acids) throughout the mesopelagic and upper bathypelagic zones. Concurrent increases in zooplankton and bacterial lipid biomarkers (e.g., 18:1ω9, odd- and branched-chain fatty acids, cholesterol) indicated that this transient pulse of labile carbon stimulated mesopelagic zooplankton and microbial activity, resulting in an enrichment of labile materials in the suspended particle pool throughout the deep ocean. Our study demonstrates that extreme weather events can have a major impact on carbon flux and particle cycling in the deep ocean, with pronounced implications for the ocean carbon pump and deep ocean ecosystems.

Tracking Fatty Acids From Phytoplankton to Jellyfish Polyps Under Different Stress Regimes: A Three Trophic Levels Experiment

The impacts of biochemicals driving food web processes are under investigation for just the last few decades. In addition, as jellyfish are drawing increasing attentions because of their mass developments and of their potential capacity of driving food web structures and energy flow by ‘top-down’ and ‘bottom-up’ controls. We here show that the provision with the biochemical complex thiamin (vitamin B1) to the common phytoplankton Rhodomonas baltica altered its fatty acid (FA) pattern towards 3-highly-unsaturated FAs (3-HUFA) and that this pattern was further transferred up to the zooplankton consumer, the copepod Acartia tonsa. However, polyps of the Jellyfish Aurelia aurita feeding on A. tonsa only had a low relative 3-HUFA content, especially due to a reduction in 22:63 (DHA), but elevated levels of 20:46 (ARA). The high proportion of the -6 HUFA, ARA in polyps may provide evidence for preferential conversion of ARA in polyps, eventually from DHA in a so far unknown pathway. In contrast to A. tonsa, newly hatched A. salina nauplii used as food for A. aurita polyps were almost devoid of HUFA, but contained high levels of C18 polyunsaturated FAs (C18-PUFA). Consequently, polyps feeding on them contained few HUFA, while high levels of C18-PUFA predominated. This suggests that A. aurita polyps cannot efficiently convert 3 C18-PUFA to 3-HUFA. In addition, besides a decrease in saturated FAs, especially an increase in HUFA in A. aurita polyps with decreasing temperature was observed, for which the dietary provision with HUFA seemed to be critical. Altering the FA pattern as a response of temperature reflects an adaptation to seasonal changes and may be related to their life history plasticity.

A non-venomous sPLA2 of a lepidopteran insect: Its physiological functions in development and immunity

Eicosanoids are oxygenated C20 polyunsaturated fatty acids that mediate various physiological processes in insects. Eicosanoid biosynthesis begins with a C20 precursor, arachidonic acid (5,8,11,14-eicosatetraenoic acid: AA). AA is usually released from phospholipids at sn-2 position by catalytic activity of phospholipase A2 (PLA2). Although various PLA2s classified into 16 gene families (= Groups) are known in various biological systems, few PLA2s are known in insects. Only two PLA2s involved in intracellular calcium independent PLA2 (iPLA2) group have been identified in lepidopteran insects with well known eicosanoid physiology. This study reports the first secretory PLA2 (sPLA2) in lepidopteran insects. A partial open reading frame (ORF) of PLA2 was obtained by interrogating Spodoptera exigua transcriptome. Subsequent 3′-RACE resulted in a full ORF (Se-sPLA2A) encoding 194 amino acid sequence containing signal peptide, calcium-binding domain, and catalytic site. Phylogenetic analysis indicated that Se-sPLA2A was clustered with other Group III sPLA2s. Se-sPLA2A was expressed in most larval instars except late last instar. Its expression was inducible by immune challenge and juvenile hormone analog injection. RNA interference of Se-sPLA2A significantly suppressed cellular immunity and impaired larval development. These results suggest that non-venomous sPLA2 plays a crucial role in immune and developmental processes in S. exigua, a lepidopteran insect.

Importance of extracellular matrix and growth state for the EA.hy926 endothelial cell response to polyunsaturated fatty acids.

Consumption of different PUFAs (polyunsaturated fatty acids) can induce functional changes in blood vessels via endothelial cells, which interact with dietary factors in the circulation. The basement membrane that separates the endothelium from the smooth muscle cells of the medial layer can also influence the functional state of endothelial cells. However, the effect of basement membrane on the endothelial response to dietary PUFAs in relation to growth state (e.g. proliferation versus quiescence) has never been investigated. We therefore compared the viability (CCK kit) and proliferation (bromodeoxyuridine incorporation) of EA.hy926 endothelial cells grown on Matrigel or collagen versus non-coated plates. EA.hy926 viability and proliferation were also assessed after treatment with 0–150 μM of PUFAs [linoleic acid (LA), arachidonic acid (AA), α-linolenic acid (ALA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)]. Our study showed that only cells grown on Matrigel-coated plates reached quiescence after becoming confluent with a decreased level of MCM2 and p-cyclin D1 (T286), increased levels of p27kip1 and a low level of apoptosis and senescence. AA, EPA and DHA decreased the viability and proliferation of subconfluent cells grown on plastic dishes in a dose-dependent manner, while the presence of Matrigel made the cells resistant to these adverse effects. Confluent cell viability was less sensitive to higher concentrations of AA, EPA and DHA than subconfluent cells, and a significant increase in caspase-3 cleavage was only observed in confluent cells treated with DHA. Higher concentrations of AA, EPA and DHA suppressed DNA synthesis by both subconfluent and confluent cells, while precursor C18 PUFAs (LA and ALA) had no negative effects on viability and proliferation. Our study is the first to show that extracellular matrix and growth state are important factors in the EA.hy926 cell response to PUFAs, and that the mechanisms by which individual PUFAs operate may be growth state-dependent.

Regiospecificity of a novel bacterial lipoxygenase from Myxococcus xanthus for polyunsaturated fatty acids

Lipoxygenase (LOX) is the key enzyme involved in the synthesis of oxylipins as signaling compounds that are important for cell growth and development, inflammation, and pathogenesis in various organisms. The regiospecificity of LOX from Myxococcus xanthus, a gram-negative bacterium, was investigated. The enzyme catalyzed oxygenation at the n-9 position in C20 and C22 polyunsaturated fatty acids (PUFAs) to form 12S- and 14S-hydroxy fatty acids (HFAs), respectively, and oxygenation at the n-6 position in C18 PUFAs to form 13-HFAs. The 12S-form products of C20 and C22 PUFAs by M. xanthus LOX is the first report of bacterial LOXs. The residues involved in regiospecificity were determined to be Thr397, Ala461, and Ile664 by analyzing amino acid alignment and a homology model based on human arachidonate 15-LOX with a sequence identity of 25%. Among these variants, the regiospecificity of the T397Y variant for C20 and C22 PUFAs was changed. This may be because of the reduced size of the substrate-binding pocket by substitution of the smaller Thr to the larger Tyr residue. The T397Y variant catalyzed oxygenation at the n-6 position in C20 and C22 PUFAs to form 15- and 17-hydroperoxy fatty acids, respectively. However, the oxygenation position of T397Y for C18 PUFAs was not changed. The discovery of bacterial LOX with novel regiospecificity will facilitate the biosynthesis of regiospecific‑oxygenated signaling compounds.

Cloning, tissue distribution, functional characterization and nutritional regulation of a fatty acyl Elovl5 elongase in chu's croaker Nibea coibor

Enzymes that lengthen the carbon chain of polyunsaturated fatty acids (PUFA) are key to the biosynthesis of the long-chain polyunsaturated fatty acids (LC-PUFA). Here we report on the molecular cloning, tissue distribution, functional characterization and nutritional regulation of a elovl5 gene from Nibea coibor. The full-length cDNA was 1315 bp, including a 5-untranslated region (UTR) of 134 bp, a 3-UTR of 296 bp and an open reading frame of 885 bp, which specified a peptide of 294 amino acids. Bioinformatics analysis showed that the deduced peptide sequence possessed all the characteristic features of microsomal fatty acyl elongases, including the so-called histidine box (HXXHH), the canonical C-terminal endoplasmic reticulum retention signal, several predicted transmembrane regions and other highly conserved motifs. Expression of elovl5 was strongly observed in stomach, and more weakly in kidney, spleen, intestine, brain, eye, liver, gill, muscle and heart. Functional characterization revealed that the chu's croaker Elovl5 was able to elongate both C18 and C20 PUFA substrates. Nutritional study indicated that the hepatic expression of elovl5 could be up-regulated by low dietary n−3 LC-PUFA. These results may contribute to better understanding the LC-PUFA biosynthetic pathway and regulation mechanism in chu's croaker.

Quantification of Fatty Acid Methyl Esters in Various Biological Matrices by LC-DAD and LC-MS after One-Step Transesterification

The investigation of lipids is a complex field and many analytical methods are applied. Gas chromatography (GC) methods are the standard methods; however, GC systems are not available for every researcher. Hence, an alternative liquid chromatography (LC) method was designed. The LC method was developed and validated for separation of C16:n and C18:n fatty acid methyl esters (FAMEs). Baseline-separation of eight FAMEs was achieved with one C18 column. Two ways of detection (diode array detector, DAD, and mass spectrometry, MS), were established and compared. The validation results proved that the method is precise (RSD 0.99) for all analysed FAMEs. Limits of detection (LOD) down to 3 ng and limits of quantification (LOQ) down to 6 ng were achieved. For sample preparation, a one-step transesterification method was used. In bacon fat, duckling lard, and butter, palmitic acid, stearic acid, and oleic acid were the main fatty acids. Olive oil had the highest oleic acid content (60% w/w). Linseed extract showed the highest linolenic acid content (60% w/w). In fungi extract, only a few fatty acids were detected. The highest fatty acid diversity was found in algae extract. Seven of the eight investigated fatty acids were found therein including C16 polyunsaturated fatty acids (PUFAs); further, uncommon C16 and C18 PUFAs were identified. In conclusion, the LC method with DAD detection represents a simple alternative to the usually used GC methods. Using MS detection, unknown compounds can be additionally identified and 10-fold lower LODs can be achieved.

Lipidomic analysis of Botryococcus (Trebouxiophyceae, Chlorophyta) - Identification of lipid classes containing very long chain fatty acids by offline two-dimensional LC-tandem MS

Very long chain fatty acids (VLCFAs) were identified in four strains of the green alga Botryococcus braunii (Trebouxiophyceae). The algae contained a series of monoenoic fatty acids up to triacontenoic acid and further VLCFAs in amounts around 1% of total fatty acids. The separation of lipid classes using hydrophilic interaction chromatography revealed that the most abundant VLCFAs (28:2, 28:1 and 28:0) were contained in neutral lipids (triacylglycerols and/or diacylglycerols) and in phospholipids (phosphatidic acid and/or phosphatidylcholine). Using non-aqueous reversed-phase liquid chromatography tandem mass spectrometry (NARP-LC/MS2) of the appropriate collected fractions, molecular species of triacylglycerols containing one or two VLCFAs were described and phosphatidylcholines containing VLCFAs were separated for the first time. Because the presence of Botryosphaerella sudetica (Chlorophyceae) as contaminant of Botryococcus braunii strain Droop 1950/807-1 placed some doubts on the results of previous studies, a strain of this green alga of was also analyzed. In contrast to Botryococcus, C16, a substantially lower proportion of C18 polyunsaturated fatty acids and no VLCFAs were detected in Botryosphaerella.

Survey of the C20 and C22 oxylipin family in marine diatoms

Diatoms are highly abundant microalgae that are prolific sources for bioactive oxylipins, which are generated by the oxidative transformation of polyunsaturated fatty acids and contribute to the chemical defense of algae. We introduce herein a targeted survey of oxylipins from four diatom species with a focus on metabolites that also occur in mammalians. Analytical data from LC–MS/MS measurements indicate that diatoms produce a surprising variety of oxylipins, including several hydroxylated and dihydroxylated C18-, C20-, and C22-polyunsaturated fatty acids. It was also found that diatoms represent the first non-mammalian source for the resolvins (Rv)E2 and RvE3, which have recently been identified as modulators of inflammation in humans and rodents. The oxylipins are produced in response to mechanical disruption of the cells, suggesting their potential involvement in the wound-activated chemical defense of these diatoms.

Elovl4 can effectively elongate C18 polyunsaturated fatty acids in loach Misgurnus anguillicaudatus

In this study, full-length cDNA sequences of elovl4a and elovl4b from loach Misgurnus anguillicaudatus were cloned. The full-length cDNAs of loach elovl4a and elovl4b were 2423 and 2054bp, encoding 315 and 300 amino acids, respectively. The deduced amino acid sequences of elovl4a and elovl4b in loach both shared the highest identity with those of Danio rerio, whereas lower identity score between loach elovl4a and elovl4b was present. Temporal expression and tissue expression of loach elovl4a and elovl4b were studied by reverse transcriptase PCR. Results of the tissue expression analyses suggested different functions of loach elovl4a and elovl4b. Functional characterizations of loach elovl4a and elovl4b on synthesis of fatty acids, especially elongating C18 polyunsaturated fatty acids (PUFAs) to longer-chain fatty acids, were studied by heterologous expression in Saccharomyces cerevisiae. Loach elovl4a and elvol4b enzymes were able to elongate all fatty acids tested including 18:2n-6, 18:3n-3, 18:3n-6, 20:4n-6 and 20:5n-3. At last, expression levels of the two elovl4 genes of loach fin cells incubated with 18:2n-6 and 18:3n-3 of different concentrations were measured. Expressions of elovl4a and elovl4b of loach fin cells were significantly up-regulated by 18:2n-6 and 18:3n-3. The results obtained here indicated that loach elovl4 could effectively elongate C18 PUFAs. This was a systematic report of elovl4's elongating functions towards C18 and provided an alternative pathway for C20 biosynthesis in fish species.

Eicosanoid-mediated immunity in insects

Eicosanoid is a collective term for oxygenated metabolites of C20 polyunsaturated fatty acids. As seen in mammals, eicosanoids play crucial roles in mediating various physiological processes, including immune responses, in insects. Upon microbial pathogen infection, non-self recognition signals are propagated to nearly immune effectors such as hemocytes and fat body using various immune mediators, in which eicosanoid signals act as the ultimate downstream mediator. The chemical diversity of eicosanoids may operate to mediate various immune responses. Some entomopathogenic bacteria suppress eicosanoid biosynthesis, which inhibits host insect immunity and promotes their pathogenicity. This review introduces immune responses mediated by various eicosanoids. Then it explains the cross-talks of eicosanoids with other immune mediators including cytokines, biogenic monoamines, and nitric oxide to clarify the complexity of insect immune mediation. Finally, we highlight the biological significance of eicosanoids by demonstrating bacterial pathogenicity inhibiting a key enzyme – phospholipase A2 – in eicosanoid biosynthesis using their secondary metabolites to defend host insect immune attack.

Silkworm (Bombyx mori) has the Capability to Accumulate C20 and C22 Polyunsaturated Fatty Acids

Bombyx mori, an insect with significant economic importance in China contains a large amount of lipids among different developmental stages, especially the pupal stage. Silkworms are fed with a base diet and fish oil supplemented diet, and the fatty acid (FA) compositions of different strains, life stages, and sexes are determined by GC‐MS. No C20 or C22 polyunsaturated fatty acids (PUFAs) are detected in silkworms fed with the base diet. Females accumulates more unsaturated fatty acids (USFAs) than males in the pupal stage, and the relative amount of USFAs decreases from the wandering stage to the moth stage, especially in females, suggesting that silkworms prefer the utilization of USFAs. In addition, when silkworms are fed with a diet supplemented with fish oil, which contained C20 and C22 PUFAs, they accumulates C20 and C22 PUFAs in proportion to the concentration of fish oil added to the diet.Practical Applications: Currently, the main dietary source of C20 and C22 PUFAs is marine fish, however, of which the stocks are declining because of environmental pollution and overfishing. Because of its excellent capability to accumulate C20 and C22 PUFAs, silkworm could be developed as a potential new source of these long‐chain FAs by transferring ?5 and ?6 desaturases to silkworm or its primary food, mulberry.The silkworm (Bombyx mori) can accumulate C20 and C22 polyunsaturated fatty acids (PUFAs) from diet supplemented with these PUFAs (B). The accumulation efficiency of eicosapentaenoic acid (EPA) in silkworm is particularly outstanding. This capacity may make silkworm a potential new source of C20 and C22 PUFAs.

Enrichment of C20-22 Polyunsaturated Fatty Acids from Refined Liver Oil of Leafscale Gulper Shark, Centrophorus squamosus

ABSTRACTThe C20-22 polyunsaturated fatty acids (C20-22 PUFAs), mainly eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), were concentrated from the refined liver oil of deep sea leafscale gulper shark, Centrophorus squamosus, by sequential processes of winterization, urea complexation, and argentation chromatography. Winterization at 4°C using acetonitrile as solvent showed significant reduction of the total saturated fatty acid content (12%) with a concomitant increase of polyunsaturated fatty acids (36.3%). The urea complexation process significantly enriched the total polyunsaturated fatty acids (49.5%), with a reduction in saturated fatty acids (83.9%). Silica gel based argentation chromatography further concentrated the esters of C20-22 PUFA (>99 percent purity). Nuclear Magnetic Resonance spectroscopy was used as a chemical fingerprinting tool to study the enrichment of C20-22 PUFAs at various stages of the purification process. An increase in the proton signal intensity at the olefinic region (at δ 4–6) and a decrease in the aliphatic signals (at δ 0.5–2) showed that the process successfully enriched the C20-22 PUFAs.Abbreviations: PUFA, polyunsaturated fatty acid; EPA, eicosapentaenoic acid; DHA, docosahexaenoic acid; FAME, fatty acid methyl esters; SFA, saturated fatty acid; MUFA, monounsaturated fatty acid; NMR, nuclear magnetic resonance; GCMS, gas chromatography mass spectrometry; FFA, free fatty acid.

Dietary supplementation of papaya (Carica papaya l.) leaf affects abundance of Butyrivibrio fibrisolvens and modulates biohydrogenation of C18 polyunsaturated fatty acids in the rumen of goats

Saturated fatty acid (SFA) produced as a final product of rumen biohydrogenation (BH) by rumen microbes is harmful to human health because of increasing the risk of heart-related diseases. The objectives of this study were to test the effect of papaya leaf (PL) on the BH of C18 polyunsaturated fatty acids (PUFA), fatty acid composition and bacterial abundance in the rumen of goats. Three rumen fistulated male goats were assigned in a 3 × 3 latin square design using three levels of PL; no addition of PL in basal diet (control, CON, 50% concentrate +50% alfalfa hay), 25% of alfalfa hay in basal diet replaced by PL (medium PL, MPL) and 50% of alfalfa hay in basal diet replaced by PL (high PL, HPL). Supplementation of the diet with PL significantly (p < .05) decreased the rate of rumen BH of C18:2n-6 (linoleic acid, LA) at different hours of sampling at the higher inclusion of PL (HPL), resulted in the reduction (p < .05) of C18:0 (stearic acid, SA) in the rumen liquor. The population of Butyrivibrio fibrisolvens as determined by the real-time PCR was higher (p < .05) for MPL (5.78 cell/mL) and HPL (6.09 cell/mL) in the rumen of goats as compared with the CON (5.50 cell/mL). The current results imply that PL supplementation could be a useful feeding strategy to modulate the BH of LA in rumen of goat.