C20 Polyunsaturated Fatty Acids Research Articles

Effects of genetic selection and threonine on meat quality in Pekin ducks

The present study was conducted to investigate the effects of genetic selection and threonine levels on meat quality in Pekin ducks. At 15 D of age, 192 lean ducks and 192 fatty ducks were selected and allotted to one of three treatments with 8 replicates with similar BW (8 ducks/cage), respectively. All ducks were fed the experimental diets (0.00, 0.15, and 0.30% added threonine) for 21 D from 15 to 35 D of age. The results showed that fatty ducks had higher (P < 0.001) feed intake, feed/gain ratio, abdominal fat percentage, and sebum percentage and lower (P = 0.001) breast muscle percentage compared with that of lean ducks. The fatty-type and lean-type ducks had similar weight gain and BW. Dietary threonine supplementation improved (P < 0.05) growth performance and increased breast muscle percentage in lean-type ducks, but it did not affect (P > 0.05) those indices in fatty-type ducks. Lean ducks had higher (P < 0.001) hepatic contents of total lipids, triglyceride, cholesterol, and plasma low-density lipoprotein cholesterol concentration, and dietary threonine supplementation decreased (P < 0.05) hepatic total lipid, cholesterol, and triglyceride contents in lean ducks, but it had no influence on hepatic lipids in fatty ducks (P > 0.05). Lean ducks had higher (P < 0.05) concentrations of monounsaturated fatty acid (MUFA), and C18-polyunsaturated fatty acid (PUFA) in the liver, PUFA in the breast muscle, and C18:3n6 and C18:3n3 in plasma and lower C20-PUFA and C22-PUFA in the liver and MUFA in plasma, compared with fatty ducks. Threonine supplementation increased PUFA, N3-PUFA, and n6-PUFA in plasma and hepatic fatty acids profiles in lean ducks (P > 0.05) but had on influence on total MUFA and total PUFA in the liver, breast muscle, and plasma in fatty ducks (P > 0.05). In conclusion, genetic selection toward meat production and threonine supplementation increases meat production and PUFA contents, which would influence eating quality, but it is benefit for human health.

Genome wide identification and functional characterization of two LC-PUFA biosynthesis elongase (elovl8) genes in rabbitfish (Siganus canaliculatus)

Elongases of very long-chain fatty acids (Elovls) catalyze the rate-limiting step of the elongation pathway that results in net two‑carbon elongation of pre-existing fatty acyl chains. As a set of crucial enzymes involved in the long-chain polyunsaturated fatty acids (LC-PUFA) biosynthesis, Elovls of fish have been investigated extensively in recent years. In the present study, we first identified two novel fish-specific elovl genes (named as elovl8a and elovl8b) from the herbivorous marine teleost rabbitfish (Siganus canaliculatus) by genomic survey and molecular cloning methods. Subsequently, their functional characteristics, tissue distribution patterns and transcriptional changes in response to different nutritional states were investigated. Full-length coding sequences of the elovl8a and elovl8b genes were 804 and 792 bp, encoding 267 and 263 amino acids, respectively. Multiple alignment, genomic synteny and phylogenetic analyses further suggested that elovl8 genes were unique to teleosts. Functional characterization by heterologous expression in yeast showed that Elovl8b could elongate C18 (18:2n-6, 18:3n-3 and 18:4n-3) and C20 (20:4n-6 and 20:5n-3) polyunsaturated fatty acids (PUFAs) to longer-chain polyunsaturated fatty acids (LC-PUFAs) whereas Elovl8a lacked this ability. In vitro, the expression of elovl8b but not elovl8a in rabbitfish hepatocytes was significantly up-regulated by incubation with 18:2n-6, 18:3n-3, 20:4n-6 and 20:5n-3, respectively. In vivo, compared with fish oil, dietary vegetable oil enriched in C18 PUFA enhanced the expression of elovl8b in rabbitfish brain, liver, intestine and gill tissues. These findings suggest that elovl8b but not elovl8a is a novel active member of the Elovl protein family involved in the LC-PUFAs biosynthesis pathway in rabbitfish, and provide novel insight into the mechanisms of LC-PUFAs biosynthesis in teleost.

Characterization and Biosynthesis of Lipids in Paulinella micropora MYN1: Evidence for Efficient Integration of Chromatophores into Cellular Lipid Metabolism.

The chromatophores found in the cells of photosynthetic Paulinella species, once believed to be endosymbiotic cyanobacteria, are photosynthetic organelles that are distinct from chloroplasts. The chromatophore genome is similar to the genomes of α-cyanobacteria and encodes about 1,000 genes. Therefore, the chromatophore is an intriguing model of organelle formation. In this study, we analyzed the lipids of Paulinella micropora MYN1 to verify that this organism is a composite of cyanobacterial descendants and a heterotrophic protist. We detected glycolipids and phospholipids, as well as a betaine lipid diacylglyceryl-3-O-carboxyhydroxymethylcholine, previously detected in many marine algae. Cholesterol was the only sterol component detected, suggesting that the host cell is similar to animal cells. The glycolipids, presumably present in the chromatophores, contained mainly C16 fatty acids, whereas other classes of lipids, presumably present in the other compartments, were abundant in C20 and C22 polyunsaturated fatty acids. This suggests that chromatophores are metabolically distinct from the rest of the cell. Metabolic studies using isotopically labeled substrates showed that different fatty acids are synthesized in the chromatophore and the cytosol, which is consistent with the presence of both type I and type II fatty acid synthases, supposedly present in the cytosol and the chromatophore, respectively. Nevertheless, rapid labeling of the fatty acids in triacylglycerol and phosphatidylcholine by photosynthetically fixed carbon suggested that the chromatophores efficiently provide metabolites to the host. The metabolic and ultrastructural evidence suggests that chromatophores are tightly integrated into the whole cellular metabolism.

Assessing Polyunsaturated Fatty Acid Essentiality in Florida Pompano

AbstractWe evaluated the growth performance and fatty acid composition of juvenile Florida Pompano Trachinotus carolinus that were fed diets containing different combinations of n‐3 and n‐6 polyunsaturated fatty acids (PUFAs) to assess the relative importance of C18 PUFAs and long‐chain (LC) PUFAs in meeting the essential fatty acid requirements of this species. Juvenile fish (mean ± SD =47.4 ± 0.6 g/fish) were fed diets containing different combinations of n‐3 and n‐6 PUFAs: a positive control containing menhaden fish oil, a negative control containing hydrogenated soybean oil, and experimental diets containing hydrogenated soybean oil amended with ethyl esters of 18:3(n‐3); 18:3(n‐3) and 18:2(n‐6); 22:6(n‐3); 22:6(n‐3) and 20:4(n‐6); or 20:5(n‐3), 22:6(n‐3), and 20:4(n‐6). After 7 weeks, weight gain, feed conversion ratio, and specific growth rate were significantly greater among fish that were fed the positive control feed in comparison with those that received the negative control feed. The addition of n‐3 and/or n‐6 PUFAs to the negative control formulation did not significantly improve performance in any case, although numeric improvements observed for weight gain and feed conversion ratio appeared to suggest some advantage from providing a complement of both n‐3 and n‐6 LC‐PUFAs in the diet. Fatty acid composition varied significantly between all tissues, generally reflecting dietary fatty acid profiles. Tissue fatty acid profiles suggest that Florida Pompano have some ability to elongate and desaturate fatty acids and may be able to survive on diets containing C18 PUFAs, but they perform optimally when provided with LC‐PUFAs directly.

Reevaluating Polyunsaturated Fatty Acid Essentiality in Channel Catfish

AbstractChannel Catfish Ictalurus punctatus reportedly require alpha‐linolenic acid (18:3[n‐3]) for normal growth. However, contradicting studies suggest that elevated dietary levels of this fatty acid might cause growth inhibition, and diets containing linoleic acid (18:2[n‐6]) alone or in combination with 18:0 and 18:1(n‐9) may support growth as well as fish oil (FO)‐based diets containing increased levels of long‐chain (LC) polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (20:5[n‐3]), docosahexaenoic acid (22:6[n‐3]), and arachidonic acid (20:4[n‐6]). Further complicating the understanding of essential fatty acid (EFA) requirements in Channel Catfish are conflicting reports suggesting that feeding LC‐PUFAs can either enhance or suppress growth. We evaluated growth performance and fatty acid composition of juvenile Channel Catfish that were fed seven diets containing n‐3 and n‐6 C18 PUFAs and/or LC‐PUFAs in different combinations: a positive control containing menhaden FO, a negative control containing hydrogenated soybean oil, and experimental diets containing hydrogenated soybean oil amended with ethyl esters of 18:3(n‐3); 18:3(n‐3) and 18:2(n‐6); 22:6(n‐3); 22:6(n‐3) and 20:4(n‐6); or 20:5(n‐3), 22:6(n‐3), and 20:4(n‐6). After 18 weeks, we observed no differences in growth or conversion efficiency. However, fish that were fed diets supplemented with C18 PUFAs and LC‐PUFAs numerically outperformed those fed the FO‐based positive control feed. Tissue fatty acid profiles generally mirrored that of the diet provided, except that saturates were underrepresented in the tissues. Channel Catfish that received the EFA‐free negative control feed or the diets supplemented with 18:3(n‐3) and 18:2(n‐6) or 20:4(n‐6) and 22:6(n‐3) developed elevated ratios of 22:5(n‐6) : 22:6(n‐3), whereas an elevated ratio of 20:3(n‐9) : 20:4(n‐6) was observed among fish that were fed the 22:6(n‐3) supplemented diet. Our findings suggest that C18 PUFA and LC‐PUFA diets appear to equally satisfy the EFA requirements of this species; however, intact dietary LC‐PUFAs should be carefully balanced to avoid antagonistic effects of surplus n‐3 LC‐PUFA on n‐6 LC‐PUFA biosynthesis.

In vitro rumen biohydrogenation of unsaturated fatty acids in tropical grass-legume rations.

Aim:The aim of this study was to evaluate the effects of various combinations of tropical grass-legume species in rations on the biohydrogenation (BH) activity of unsaturated fatty acids (FAs), C18:0 composition, and fermentation profile in an in vitro rumen system.Materials and Methods:Samples of the following five fodder plants were used: One species of grass (Pennisetum purpureum) and four species of tree legumes (Leucaena leucocephala, Gliricidia sepium, Calliandra calothyrsus, and Indigofera zollingeriana). The following eight experimental diets were evaluated: 50% P. purpureum + 50% L. leucocephala (LL I); 50% P. purpureum + 50% G. sepium (GS I); 50% P. purpureum + 50% C. calothyrsus (CC I); 50% P. purpureum + 50% I. zollingeriana (IZ I); 75% P. purpureum + 25% L. leucocephala (LL II); 75% P. purpureum + 25% G. sepium (GS II); 75% P. purpureum + 25% C. calothyrsus (CC II); and 75% P. purpureum + 25% I. zollingeriana (IZ II). Each ration was replicated 3 times. In vitro rumen incubation was performed for 48 h, according to the Tilley and Terry method. Determination of the FA profiles of the forage materials and rumen fluid samples was performed using gas chromatography.Results:The percentage of polyunsaturated FA (PUFA) in the forage materials ranged from 34.18% (P. purpureum) to 74.51% (C. calothyrsus). The percentage of monounsaturated FA (MUFA) ranged from 5.06% (P. purpureum) to 8.71% (L. leucocephala). The percentage of saturated FA (SFA) was the lowest at 19.12% (C. calothyrsus) and highest at 60.76% (P. purpureum). In vitro BH of C18:3 n-3, C18:2 n-6, C18:1 n-9, and C18 PUFA in the experimental diets ranged from 72% to 100%. The BH of C18:1 n-9 in GS I (80%) and IZ I (72%) was significantly different (p<0.05). The percentage of C18:0 was 10-50% and significantly different (p<0.05) among treatments, with the highest (of 50%) in GS II. No significant differences (p>0.05) were observed in the fermentation parameters (pH, total volatile FAs, in vitro dry matter digestibility, and in vitro organic matter digestibility) among the treatments, except in NH3 concentration (p<0.05).Conclusion:The various combinations of tropical legumes do not have significant inhibitory effects on the BH of C18:2 n-6, C18:3 n-3, and C18 PUFA after in vitro incubation for 48h. Furthermore, an increase in the tropical legume ratio in the ration tends to suppress C18:0 formation after the fermentation simulation process. IZ I has the potential to reduce C18:1 n-9 (MUFA) disappearance and yield an ideal rumen fermentation profile.

Reevaluating Polyunsaturated Fatty Acid Essentiality in Hybrid Striped Bass

AbstractUnderstanding fatty acid requirements in multiple taxa is necessary to determine the degree to which dietary fish oil can be reduced or replaced with less costly, more abundant lipid sources. The polyunsaturated fatty acid (PUFA) requirements of hybrid Striped Bass (White Bass Morone chrysops × Striped Bass M. saxatilis) are currently reported as 0.5–1.0% eicosapentaenoic acid (20:5n‐3) and/or docosahexaenoic acid (22:6n‐3). This requirement does not account for n‐3 PUFA essentiality on an individual fatty acid basis and does not address possible requirements for n‐6 PUFAs (i.e., linoleic acid [18:2n‐6] or arachidonic acid [20:4n‐6]). Accordingly, hybrid Striped Bass were fed diets containing selected individual n‐3 and n‐6 C18 PUFAs, n‐3 and n‐6 long‐chain (LC) PUFAs, or combinations thereof to determine the essentiality or expendability of these nutrients. These experimental feeds were compared to feeds containing only saturated fatty acids (negative control) or menhaden fish oil (positive control). Growth performance and ratios of 22:5n‐6 : 22:6n‐3 observed in tissue fatty acid profiles suggested that dietary provision of 22:6n‐3 was adequate to satisfy physiological demand for LC‐PUFAs. However, reductions in feed conversion ratio and numeric improvements in growth suggest that dietary provision of 20:4n‐6 is also important. Results of this study indicate that provision of C18 PUFAs alone is insufficient to avoid essential fatty acid deficiency and to support optimal growth of this fish. To ensure that essential fatty acid requirements are met and performance is optimized, diets for hybrid Striped Bass should be formulated to contain 22:6n‐3 and 20:4n‐6, if not a full complement of n‐3 and n‐6 LC‐PUFAs.

Reevaluating Polyunsaturated Fatty Acid Essentiality in Nile Tilapia

AbstractNile Tilapia Oreochromis niloticus have a reported dietary requirement for the n‐6 C18 polyunsaturated fatty acid (PUFA) 18:2n‐6 (linoleic acid), but direct provision of the n‐6 long‐chain (LC) PUFA 20:4n‐6 (arachidonic acid) may be a more efficient approach. No quantitative requirement for n‐3 fatty acids has been formally recognized for tilapias, but beneficial effects of having both n‐3 and n‐6 PUFAs in the diet have been reported. To better understand PUFA essentiality in tilapias, we formulated diets to assess the growth performance and tissue fatty acid composition of juvenile Nile Tilapia (mean ± SD = 25.7 ± 0.2 g/fish) that were fed different combinations of n‐3 and n‐6 PUFAs: a positive control containing menhaden fish oil, a negative control containing hydrogenated soybean oil, and experimental diets containing hydrogenated soybean oil amended with ethyl esters of 18:3n‐3; 18:3n‐3 and 18:2n‐6; 22:6n‐3; 22:6n‐3 and 20:4n‐6; or 20:5n‐3, 22:6n‐3, and 20:4n‐6. After 7 weeks, weight gain, feed conversion ratio, and specific growth rate were significantly greater among fish that received the positive control feed than among fish that were fed the negative control feed. The addition of n‐3 and n‐6 PUFA supplements generally improved performance in most cases. No fish showed any gross indications of fatty acid deficiency, although significant differences in fatty acid deficiency indicator ratios (i.e., 20:3n‐9 : 20:4n‐6 and 22:5n‐6 : 22:6n‐3) were present in all tissues analyzed. Nile Tilapia that were fed an n‐3 PUFA‐free diet exhibited significantly elevated 22:5n‐6 : 22:6n‐3 ratios, indicating a developing n‐3 PUFA deficiency. Feeding n‐3 PUFAs—as 18:3n‐3, 22:6n‐3, or a combination of n‐3 LC‐PUFAs—had a corrective effect on these ratios, affirming the importance of a proper balance of n‐3 and n‐6 PUFAs to satisfy the essential fatty acid requirements of Nile Tilapia and further illustrating the ability of this species to satisfy physiological demand for LC‐PUFAs through de novo biotransformation of C18 PUFAs.

Rational Engineering of Hydratase from Lactobacillus acidophilus Reveals Critical Residues Directing Substrate Specificity and Regioselectivity.

Enzymatic conversion of fatty acids (FAs) by fatty acid hydratases (FAHs) presents a green and efficient route for high-value hydroxy fatty acid (HFA) production. However, limited diversity was achieved among HFAs, to date, with respect to chain length and hydroxy position. In this study, two highly similar FAHs from Lactobacillus acidophilus were compared: FA-HY2 has a narrow substrate scope and strict regioselectivity, whereas FA-HY1 utilizes longer chain substrates and hydrates various double-bond positions. It is revealed that three active-site residues play a remarkable role in directing substrate specificity and regioselectivity of hydration. If these residues on FA-HY2 are mutated to the corresponding ones in FA-HY1, a significant expansion of substrate scope and a distinct enhancement in hydration of double bonds towards the ω-end of FAs is observed. A three-residue mutant of FA-HY2 (TM-FA-HY2) displayed an impressive reversal of regioselectivity towards linoleic acid, shifting the ratio of the HFA regioisomers (10-OH/13-OH) from 99:1 to 12:88. Notable changes in regioselectivity were also observed for arachidonic acid and for C18 polyunsaturated fatty acid substrates. In addition, TM-FA-HY2 converted eicosapentaenoic acid into its 12-hydroxy product with high conversion at the preparative scale. Furthermore, it is demonstrated that microalgae are a source of diverse FAs for HFA production. This study paves the way for tailor-made FAH design to enable the production of diverse HFAs for various applications from the polymer industry to medical fields.

Fatty acid profile of duodenal digesta and of meat of feedlot beef cattle fed diets containing different levels of concentrate

The objective of this study was to evaluate the fatty acid profile of duodenal digesta (experiment I) and of meat of beef cattle (experiment II) fed diets containing different levels of concentrate (220, 400, 590 and 790 g of concentrate/kg of dry matter of the diets). The experiment I was conducted with four Charolais-Nellore steers (460 ± 18.2 kg of BW), with a T-shaped duodenal cannula, using a double 4 × 4 Latin square as an experimental design. In experiment II, 16 crossbred Charolais-Nellore young bulls (192.44 ± 18.2 kg of BW) were randomly distributed in the experimental treatments (220, 400, 590 and 790 g of concentrate/kg of dry matter of the diets). The diets were isonitrogenous (120 g of crude protein/kg of dry matter). The intramuscular fat content was used as a covariant for the statistical analysis of the meat fatty acid profile. The duodenal content of fatty acid C17:0 decreased with increase of concentrate levels, while its content in the meat presented a quadratic variation with the increase of the concentrate levels of the diets, being the lowest values observed for the diet with 400 g of concentrate. The duodenal content of fatty acid C18:1 trans-11 decreased, whereas the content of this fatty acid in the meat increased with the increase of the dietary concentrate levels. The increase in the level of concentrate reduced the content of polyunsaturated fatty acids C18:3 n-3, C20:3 n-6, C20:4 n-6, and C20:5 n-3 EPA in both the duodenal digesta and meat. No difference was observed in the n-6/n-3 fatty acids ratio (mean of 13.96) of the meat between diets. The elevation of the level of concentrate in confinement diets reduces the nutraceutical quality of the meat of Charolais-Nellore young bulls slaughtered at 14-16 months of age due to the reduction of the polyunsaturated fatty acids content important for human health.

Distinct characteristics of the substrate binding between highly homologous catalase-related allene oxide synthase and hydroperoxide lyase

A catalase-related allene oxide synthase (cAOS) or a hydroperoxide lyase (cHPL) fused together with an 8R-lipoxygenase is involved in the stress signaling of corals via an arachidonic acid pathway. cAOS gives rise to α-ketol and cyclopentenone, while cHPL catalyzes the cleavage of 8R-hydroperoxyeicosatetraenoic acid (8R-HpETE) to C8-oxo acid and C12 aldehyde. In silico analysis of the substrate entry sites of highly identical coral cAOS and cHPL indicated that two positively charged residues of cAOS, K60 and K107, and the corresponding residues of cHPL, E60 and K107, may be involved in the anchoring of the carboxy group of polyunsaturated fatty acid (PUFA) hydroperoxides. A mutational analysis of cAOS and cHPL revealed that K60 or E60 and K107 were not necessary in the tethering of 8R-HpETE, however, the E60 of cHPL was essential in the productive binding of PUFA hydroperoxides. The substrate preferences of cAOS and cHPL were determined with hydroperoxy derivatives of C18, C20, C22 PUFAs, anandamide (AEA), 1-arachidonoyl glycerol (1-AG) and selected methylated substrates. Although cAOS and cHPL were able to metabolize different free PUFA substrates and arachidonoyl derivatives, only cHPL catalyzed the reaction with methylated PUFA hydroperoxides. The differences in the substrate binding and preferences between cAOS and cHPL can be explained by the distinct properties of their substrate entry sites. The current study demonstrated that homologous PUFA metabolizing enzymes may contribute to the versatile usage of the substrate pool.

Why most insects have very low proportions of C20 polyunsaturated fatty acids: The oxidative stress hypothesis.

Eicosanoids, a group of C20 oxygenated polyunsaturated fatty acids (PUFAs), mediate various physiological processes, such as immunity, reproduction, excretion, and metabolism in insects. Arachidonic acid (AA) is used for the main precursor for the production of various eicosanoids. However, most terrestrial insects possess relatively low AA levels. Insects are presumed to be evolved sincethe Paleozoic era, at which oxygen levels might be much higher than current conditions. Compared with other animals, they exhibit relatively high metabolic rates with the well-developed tracheal system, which directly supply enough oxygen to active tissues like flight muscles. This might allow insects to be susceptible to reactive oxygen species (ROS) generated from high oxidative catabolism. Long-chain PUFAs including AA is usually reacted with ROS and become peroxidized. Peroxidized PUFAs cause various cellular damage. Thus, we propose a hypothesis that terrestrial insects minimize AA levels to minimize oxidative stress.

Effects of Supplementation of Branches and Leaves Trimmed from Tea Plant on Growth Performance, Rumen Fermentation and Meat Composition of Nanjiang Yellow Goats.

Simple SummaryThe effects of supplementation of branches and leaves trimmed from tea tree (BLTT) on growth performance, rumen fermentation characteristics and meat composition of fattening Nanjiang Yellow goats were studied. Supplementation of 4% BLTT increased final body weight of goats and also increased the activity of superoxide dismutase, while glutathione peroxidase and malondialdehyde followed the quadratic curve. Quadratic curves were also observed for villus height, crypt depth and the ratio of villus height to crypt depth in the jejunum. The quadratic effect was obtained for total essential amino acids, and individual amino acids threonine and leucine. Supplementation of 4% BLTT decreased the saturated fatty acid C16:0, and a quadratic effect was observed for polyunsaturated fatty acid C18:3 (n−3). From the present study, it is concluded that BLTT supplementation has a positive effect on body antioxidative status, gastrointestinal development, rumen fermentation characteristics and overall hence growth performance and meat composition in goats.Thirty-two 6-month-old, healthy Nanjiang Yellow goats (34.6 ± 4.16 kg) were randomly divided into four treatments to evaluate the supplementary effects of branches and leaves trimmed from tea tree (BLTT) on growth performance, rumen fermentation characteristics, and meat composition in fattening goats. The control goats were fed a basal diet. Treatments 1, 2 and 3 were fed 2%, 4% and 6% of BLTT respectively. After a 60 d feeding trial, blood samples were collected for the analysis of the antioxidant profile and goats were slaughtered to obtain the rumen fluid and carcass samples for determination of rumen fermentation characteristics and meat composition perameters. Increased final body weight was observed in goats fed 4% BLTT compared with control. The activity of superoxide dismutase was increased in the 4% BLTT-treated group, while glutathione peroxidase and malondialdehyde followed the quadratic curve. Quadratic curves were also observed for villus height, crypt depth and the ratio of villus height to crypt depth in the jejunum. The quadratic effect was obtained for total essential amino acids, and individual amino acids threonine and leucine. The saturated fatty acid C16:0 was decreased with 4% of BLTT supplementation, and a quadratic effect was observed for polyunsaturated fatty acid C18:3 (n−3). To sum up, our findings revealed that BLTT supplementation has a positive effect on body antioxidative status, gastrointestinal development, rumen fermentation characteristics and overall growth performance and meat quality in goats.

Conversion of rainforest to oil palm and rubber plantations alters energy channels in soil food webs.

In the last decades, lowland tropical rainforest has been converted in large into plantation systems. Despite the evident changes above ground, the effect of rainforest conversion on the channeling of energy in soil food webs was not studied. Here, we investigated community‐level neutral lipid fatty acid profiles in dominant soil fauna to track energy channels in rainforest, rubber, and oil palm plantations in Sumatra, Indonesia. Abundant macrofauna including Araneae, Chilopoda, and Diplopoda contained high amounts of plant and fungal biomarker fatty acids (FAs). Lumbricina had the lowest amount of plant, but the highest amount of animal‐synthesized C20 polyunsaturated FAs as compared to other soil taxa. Mesofauna detritivores (Collembola and Oribatida) contained high amounts of algal biomarker FAs. The differences in FA profiles between taxa were evident if data were analyzed across land‐use systems, suggesting that soil fauna of different size (macro‐ and mesofauna) are associated with different energy channels. Despite that, rainforest conversion changed the biomarker FA composition of soil fauna at the community level. Conversion of rainforest into oil palm plantations enhanced the plant energy channel in soil food webs and reduced the bacterial energy channel; conversion into rubber plantations reduced the AMF‐based energy channel. The changes in energy distribution within soil food webs may have significant implications for the functioning of tropical ecosystems and their response to environmental changes. At present, these responses are hard to predict considering the poor knowledge on structure and functioning of tropical soil food webs.

Biosynthetic pathway of arachidonic acid in Spodoptera exigua in response to bacterial challenge

Eicosanoids play crucial roles in mediating insect immune responses. In vertebrates, phospholipase A2 (PLA2) releases arachidonic acid (AA) from phospholipids (PLs) for biosynthesis of various eicosanoids. However, little AA is found in PLs of lepidopteran insects. Spodoptera exigua, a lepidopteran insect, is known to use eicosanoids to mediate immunity. Although AA was not detected in PLs of hemocytes and fat body (two immune tissues) of naïve larvae, it was detected at small but significant level after bacterial infection, suggesting induction of AA biosynthesis for immunity. Based on a mammalian AA biosynthetic pathway, this study hypothesizes that AA is synthesized from C18 polyunsaturated fatty acid (PUFA) precursor by subsequent desaturation and elongation reactions because PLs of S. exigua larvae are rich in linoleic acid. After inhibiting PLA2 activity to prevent release of free fatty acids, different PUFA precursors were injected to S. exigua larvae followed by assessment of eicosanoid-mediated cellular immune response. ω-6 PUFAs were effective in inducing immune response whereas α-linolenic acid (an ω-3 PUFA) was not. Several fatty acyl desaturases (SeDESs) have been predicted from S. exigua transcriptomes. Specific inhibitors against Δ5 or Δ6 DESs inhibited eicosanoid-mediated immune responses. Furthermore, RNA interference (RNAi) specific to Δ5 or Δ6 DES genes significantly suppressed eicosanoid-mediated immune responses. Four very long chain fatty acid elongase genes (SeEloV-A ∼ SeEloV-D) were predicted. Among respective RNAi treatments of these genes, only one RNAi treatment specific to type 5 elongase (SeEloV-B) suppressed eicosanoid-mediated immune response. These results suggest that S. exigua larvae can synthesize AA from linoleic acid via Δ5- and Δ6-desaturations by SeDESs along with chain elongation by SeEloV-B. Finally, this study showed significant fitness cost of uncontrolled AA biosynthesis. AA injection alone without bacterial challenge significantly induced both cellular and humoral immune responses. This unnecessary energy expense due to free AA resulted in reduced pupal size and decreased adult egg production. The detrimental effect of free AA explains physiological significance of little AA content in lepidopteran insects except for life-or-death situation such as pathogen infection.

Fecal Microbiota and Its Correlation With Fatty Acids and Free Amino Acids Metabolism in Piglets After a Lactobacillus Strain Oral Administration.

Lactobacillus has a positive effect on the host intestinal microbiota. In piglets, dietary supplementation with Lactobacillus affects general health and plays an important role in nutrient digestion and fermentation. However, this association requires further investigation. Here, we studied newborn piglets from 12 litters. The nursed piglets were given a creep feed beginning on day 10 post-partum and weaned at day 30. Piglets were fed either a control basic diet or a diet including supplementation with Lactobacillus reuteri ZLR003 at 6.0 × 106 CFU/g feed. At day 30 and 60, feces samples were taken and used for sequencing of the V3-V4 hypervariable region of the 16S rRNA gene. At day 60, feces samples and serum samples were also taken and used to measure the short chain fatty acids (SCFAs) and to detect long chain fatty acids (LCFAs) and free amino acids (FAAs), respectively. The results revealed that L. reuteri ZLR003 could improve piglet fecal microbiota composition, especially at the end of weaned period. The concentrations of lactic acid and butyric acid in feces were higher, and acetic acid concentration was lower in the L. reuteri ZLR003 group compared with the control group (P < 0.05). The serum polyunsaturated fatty acids C18:2n6c, C18:3n3, C20:4n6, and C22:6n3 were significantly higher (P < 0.05), as were the serum FAAs Gly, Ala, Val, Iso, Asn, Asp, Glu, Met, Phe, and Leu (P < 0.05), in the L. reuteri group compared with the control group. A correlation analysis revealed that the genera Ruminococcaceae_UCG-010 and Ruminococcaceae_UCG-014 had a negative correlation with the SCFAs content in feces, the genus Prevotella_9 had a higher positive correlation with C18:2n6c, and the genera Megasphaera and Mitsuokella had a more positive significant effect on the serum FAAs content in weaned piglets in the L. reuteri ZLR003 group compared with the control group. In conclusion, L. reuteri ZLR003 influenced the fecal microbiota composition of piglets, and its effects were related to the metabolism of SCFAs, LCFAs, and FAAs. Our findings will help facilitate the application of Lactobacillus strains in pig production.

Elovl4a participates in LC-PUFA biosynthesis and is regulated by PPAR\u03b1\u03b2 in golden pompano Trachinotus ovatus (Linnaeus 1758)

The elongases of very long-chain fatty acids (Elovls) are responsible for the rate-limiting elongation process in long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis. The transcription factor, PPARα, regulates lipid metabolism in mammals; however, the detailed mechanism whereby PPARαb regulates Elovls remains largely unknown in fish. In the present study, we report the full length cDNA sequence of Trachinotus ovatus Elovl4a (ToElovl4a), which encodes a 320 amino acid polypeptide that possesses five putative membrane-spanning domains, a conserved HXXHH histidine motif and an ER retrieval signal. Phylogenetic analysis revealed that the deduced protein of ToElovl4a is highly conserved with the Oreochromis niloticus corresponding homologue. Moreover, functional characterization by heterologous expression in yeast indicated that ToElovl4a can elongate C18 up to C20 polyunsaturated fatty acids. A nutritional study showed that the protein expressions of ToElovl4a in the brain and liver were not significantly affected among the different treatments. The region from PGL3-basic-Elovl4a-5 (−148 bp to +258 bp) is defined as the core promoter via a progressive deletion mutation of ToElovl4a. The results from promoter activity assays suggest that ToElovl4a transcription is positively regulated by PPARαb. Mutation analyses indicated that the M2 binding site of PPARαb is functionally important for protein binding, and transcriptional activity of the ToElovl4a promoter significantly decreased after targeted mutation. Furthermore, PPARαb RNA interference reduced ToPPARαb and ToElovl4a expression at the protein levels in a time-dependent manner. In summary, PPARαb may promote the biosynthesis of LC-PUFA by regulating ToElovl4a expression in fish.

Microbial Synthesis of Linoleate 9 S-Lipoxygenase Derived Plant C18 Oxylipins from C18 Polyunsaturated Fatty Acids.

Plant oxylipins, including hydroxy fatty acids, epoxy hydroxy fatty acids, and trihydroxy fatty acids, which are biosynthesized from C18 polyunsaturated fatty acids (PUFAs), are involved in pathogen-specific defense mechanisms against fungal infections. However, their quantitative biotransformation by plant enzymes has not been reported. A few bacteria produce C18 trihydroxy fatty acids, but the enzymes and pathways related to the biosynthesis of plant oxylipins in bacteria have not been reported. In this study, we first report the biotransformation of C18 PUFAs into plant C18 oxylipins by expressing linoleate 9 S-lipoxygenase with and without epoxide hydrolase from the proteobacterium Myxococcus xanthus in recombinant Escherichia coli. Among the nine types of plant oxylipins, 12,13-epoxy-14-hydroxy- cis, cis-9,15-octadecadienoic acid was identified as a new compound by NMR analysis, and 9,10,11-hydroxy- cis, cis-6,12-octadecadienoic acid and 12,13,14-trihydroxy- cis, cis-9,15-octadecadienoic were suggested as new compounds by LC-MS/MS analysis. This study shows that bioactive plant oxylipins can be produced by microbial enzymes.

Molecular cloning and functional characterization of elongase (elovl5) and fatty acyl desaturase (fads2) in sciaenid, Nibea diacanthus (Lacepède, 1802)

In the present paper, we investigated the molecular cloning and functional characterization of elongase of very long chain fatty acid (elovl) and fatty acyl desaturase (fads) genes in a marine teleost, Nibea diacanthus. The elongase cDNA sequence encoded a polypeptide of 294 amino acids exhibiting Elovl5 activity, which effectively elongated both C18 (18:2n−6, 18:3n−3 and 18:3n−6) and C20 (20:4n−6 and 20:5n−3) polyunsaturated fatty acids. The desaturase cDNA sequence specified a polypeptide of 445 amino acids indicating Δ6 desaturation activity, which coul converted C18:2n−6 and C18:3n−3 to C18:3n−6 and C18:4n−3, respectively. Tissue distribution analysis by quantitative real-time PCR revealed that the elovl5 was primarily expressed in intestine and liver, while the fads2 was mainly expressed in liver and brain. These results increase our knowledge of the ability of endogenous highly unsaturated fatty acids (HUFA) biosynthesis in marine carnivorous fish.

Effects of soybean oil supplement to diets of lactating dairy cows, on productive performance, and milk fat acids profile: a meta-analysis

The aim of this paper was to investigate the effects of soybean oil, an oilseed rich in the polyunsaturated fatty acid C18:2, on dry matter intake (DMI), milk yield (MY) and milk composition (including fatty acid profile) of dairy cows by using a meta-analysis approach. In the meta-analysis, effect size for all outcomes was reported as standardised means difference with 95% confidence intervals. Q test and I2 were calculated to detect the heterogeneity, with a meta-regression also used to investigate sources of heterogeneity. Results of the meta-analysis indicated that adding soybean oil to the diet of dairy cows decreased DMI, milk fat percentage (MFP), milk fat yield (MFY) and milk protein percentage (MPP), whilst significantly increasing MY. Unsaturated fatty acids C18:2, C18:1 trans-11 and C18:2 cis-9 trans-11 conjugated linoleic acid (CLA) all significantly increased with soybean oil inclusion in the diet, whereas C18:3 was numerically higher but not significantly different. Milk composition, including MFP, MFY, MPP, short-chain and medium-chain fatty acids (C4–C20) and unsaturated fatty acids showed a significant percentage of heterogeneity. Results of the meta-regression indicated that dose of soybean oil, total fatty acids in the diet, and the concentration of C18:2 between groups receiving soybean oil and control are the most important factors affecting this heterogeneity. By performing meta-analysis and meta-regression it can be concluded that dietary supplementation of soybean oil to lactating dairy cows decreased DMI, and increased MY and the amount of unsaturated fatty acids in milk.HighlightsSoybean oil dietary supplementation to dairy cows increased milk yield but reduced milk fat content.Milk fat in soybean oil dietary supplemented cows has a lower content of short-chain fatty acids, associated with de novo synthesis.Milk fat in soybean oil dietary supplemented cows has a higher content of C18:1 trans-11, C18:2 and CLA.