n-3 Canola oil effectively replaces fish oil as a new safe dietary source of DHA in feed for juvenile Atlantic salmon.

Limited availability of fish oils (FO), rich in n-3 long-chain (≥C20) PUFA, is a major constraint for further growth of the aquaculture industry. Long-chain n-3 rich oils from crops GM with algal genes are promising new sources for the industry. This project studied the use of a newly developed n-3 canola oil (DHA-CA) in diets of Atlantic salmon fingerlings in freshwater. The DHA-CA oil has high proportions of the n-3 fatty acids (FA) 18 : 3n-3 and DHA and lower proportions of n-6 FA than conventional plant oils. Levels of phytosterols, vitamin E and minerals in the DHA-CA were within the natural variation of commercial canola oils. Pesticides, mycotoxins, polyaromatic hydrocarbons and heavy metals were below lowest qualifiable concentration. Two feeding trials were conducted to evaluate effects of two dietary levels of DHA-CA compared with two dietary levels of FO at two water temperatures. Fish increased their weight approximately 20-fold at 16°C and 12-fold at 12°C during the experimental periods, with equal growth in salmon fed the FO diets compared with DHA-CA diets. Salmon fed DHA-CA diets had approximately the same EPA+DHA content in whole body as salmon fed FO diets. Gene expression, lipid composition and oxidative stress-related enzyme activities showed only minor differences between the dietary groups, and the effects were mostly a result of dietary oil level, rather than the oil source. The results demonstrated that DHA-CA is a safe and effective replacement for FO in diets of Atlantic salmon during the sensitive fingerling life-stage.

Enhancing Omega-3 Long-Chain Polyunsaturated Fatty Acid Content of Dairy-Derived Foods for Human Consumption.

Omega-3 polyunsaturated fatty acids (n-3 PUFA) are termed essential fatty acids because they cannot be synthesized de novo by humans due to the lack of delta-12 and delta-15 desaturase enzymes and must therefore be acquired from the diet. n-3 PUFA include α-linolenic acid (ALA, 18:3n-3), eicosapentaenoic (EPA, 20:5n-3), docosahexaenoic (DHA, 22:6n-3), and the less recognized docosapentaenoic acid (DPA, 22:5n-3). The three long-chain (≥C20) n-3 PUFA (n-3 LC-PUFA), EPA, DHA, and DPA play an important role in human health by reducing the risk of chronic diseases. Up to the present time, seafood, and in particular, fish oil-derived products, have been the richest sources of n-3 LC-PUFA. The human diet generally contains insufficient amounts of these essential FA due largely to the low consumption of seafood. This issue provides opportunities to enrich the content of n-3 PUFA in other common food groups. Milk and milk products have traditionally been a major component of human diets, but are also among some of the poorest sources of n-3 PUFA. Consideration of the high consumption of milk and its processed products worldwide and the human health benefits has led to a large number of studies targeting the enhancement of n-3 PUFA content in dairy products. The main objective of this review was to evaluate the major strategies that have been employed to enhance n-3 PUFA content in dairy products and to unravel potential knowledge gaps for further research on this topic. Nutritional manipulation to date has been the main approach for altering milk fatty acids (FA) in ruminants. However, the main challenge is ruminal biohydrogenation in which dietary PUFA are hydrogenated into monounsaturated FA and/or ultimately, saturated FA, due to rumen microbial activities. The inclusion of oil seed and vegetable oil in dairy animal diets significantly elevates ALA content, while the addition of rumen-protected marine-derived supplements is the most effective way to increase the concentration of EPA, DHA, and DPA in dairy products. In our view, the mechanisms of n-3 LC-PUFA biosynthesis pathway from ALA and the biohydrogenation of individual n-3 LC-PUFA in ruminants need to be better elucidated. Identified knowledge gaps regarding the activities of candidate genes regulating the concentrations of n-3 PUFA and the responses of ruminants to specific lipid supplementation regimes are also critical to a greater understanding of nutrition-genetics interactions driving lipid metabolism.

Correlations between growth and wool quality traits of genetically divergent Australian lambs in response to canola or flaxseed oil supplementation.

The correlations between growth and wool traits in response to canola and flaxseed oil supplementation were evaluated in Australian prime lambs. Sixty dual-purpose prime lambs including purebred Merino and crossbred lambs were allocated to one of five treatments of lucerne hay basal diet supplemented with isocaloric and isonitrogenous wheat-based pellets. Treatments were: no oil inclusion (Control); 2.5% canola oil; 5% canola oil; 2.5% flaxseed oil and 5% flaxseed oil, with lamb groups balanced by breed and gender. Each lamb was daily supplemented with 1kg of pellets and had free access to lucerne hay and water throughout the 7-week feeding trial, after a 3-week adaptation. Individual animal basal and supplementary pellet feed intakes were recorded daily, while body conformation traits, body condition scores and liveweights were measured on days 0, 21, 35 and 49. The lambs were dye-banded on the mid-side and shorn before commencing the feeding trial and mid-side wool samples were collected from the same dye-banded area of each lamb at the end of the experiment. Correlations between wool quality traits and lamb performance were non-significant (P>0.05). Oil supplementation had no detrimental effect on lamb growth and wool quality traits (P > 0.05). Gender significantly affected wither height gain and fibre diameter. There were significant interactions between oil supplementation and lamb breed on chest girth. The correlations between clean fleece yield (CFY) and other wool quality traits were moderate ranging from 0.29 to 0.55. Moderate to high correlations between fibre diameter (FD) and other wool quality traits were detected (0.46-0.99) with the strongest relationship between FD and wool spinning fineness (SF). The relationship between CFY and wool comfort factor (CF) were positive, while negative relationships between CFY and the others were observed. A combination of 5% oil supplementation and genetics is an effective and strategic management tool for enhancing feed efficiency and growth performance without negative effects on wool quality in dual-purpose lamb production. This is a good outcome for dual-purpose sheep farmers. It essentially means the absorbed nutrients in supplemented lambs yielded good growth performance without any detrimental impact on wool quality; a win-win case of nutrient partitioning into the synthesis of muscle and wool without compromising either traits.

Enhancement of dairy sheep cheese eating quality with increased n-3 long-chain polyunsaturated fatty acids.

This study investigated the effect of different plant oil-infused and rumen-protected wheat-based pellets containing eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) on n-3 long-chain (≥C20) polyunsaturated fatty acid (LC-PUFA) content, fatty acid recovery, and sensory attributes of ripened cheese from dairy sheep. During a 10-wk supplementary feeding trial, 60 dairy ewes balanced by live weight, milk yield, parity, and sire breed were randomly divided into 6 groups that were (1) supplemented with on-farm existing commercial wheat-based pellets without oil inclusion (control) or supplemented with wheat-based pellets infused with 50 mL/kg dry matter of oils from (2) canola, (3) rice bran, (4) flaxseed, (5) safflower, and (6) rumen-protected EPA + DHA. Milk samples from each treatment were collected separately by sire breed during the experimental period for cheese processing at the end of the experiment. Twelve batches of cheese (2 batches per treatment) were processed and ripened for 120 d. Three cheese samples were collected and analyzed for each cheese making session (total of 36 cheese samples) at d 120 of ripening. Processed cheese of rumen-protected EPA + DHA had the most efficiency at elevating total n-3 LC-PUFA [total EPA + DHA + docosapentaenoic acid (DPA, 22:5n-3] content compared with the control (0.49 vs. 0.28%). Flaxseed elicited the greatest enhancement of α-linolenic acid (ALA, 18:3n-3), whereas safflower was the most effective diet in enhancing the level of linoleic acid (18:2n-6) in cheese (1.29 vs. 0.71% and 4.8 vs. 3.3%, respectively). Parallel recoveries of n-3 and n-6 LC-PUFA were observed across all treatments except for α-linolenic acid and EPA. Cheese eating sensory traits were also highly affected by oil supplementation, with the highest score of 7.5 in cheese from the rice bran and flaxseed treatments. These results provide new insights into the biological mechanisms and processes that determine dairy ewe milk productivity by underpinning the vital biological role of n-3 LC-PUFA in not only enhancing the healthy composition of cheese from ewes but also translating it into consumer acceptability.

Supplementing Dairy Ewes Grazing Low Quality Pastures with Plant-Derived and Rumen-Protected Oils Containing Eicosapentaenoic Acid and Docosahexaenoic Acid Pellets Increases Body Condition Score and Milk, Fat, and Protein Yields.

The Australian dairy sheep industry is small and mostly based on a natural grass grazing system, which can limit productivity. The current study tested different plant oil-infused and rumen protected polyunsaturated fats and their interactions with sire breeds to improve lactation traits and body condition scores (BCS) of ewes grazing low quality pastures. It was hypothesised that supplementing lactating ewe's diets with plant-derived polyunsaturated oils would improve milk production and composition without compromising BCS. Sixty ewes (n = 10/treatment) in mid-lactation, balanced by sire breed, parity, milk yield, body condition score, and liveweight, were supplemented with: (1) control: wheat-based pellets without oil inclusion; wheat-based pellets including; (2) canola oil (CO); (3) rice bran oil (RBO); (4) flaxseed oil (FSO); (5); safflower oil (SFO); and (6) rumen protected marine oil containing eicosapentaenoic acid and docosahexaenoic acid (RPO). Except for the control group, all supplementary diets included the same level of 50 mL/kg DM of oil and all diets were isocaloric and isonitrogenous. Experimental animals were grazed in the same paddock with ad libitum access to pasture, hay, and water during the 10-week study. RPO was the most effective diet that enhanced milk, fat, and protein yields by approximately 30%, 13%, and 31%, respectively (p < 0.0001). A significant increase in milk production was also observed with CO, RBO, and SFO treatments (p < 0.0001). Breed significantly influenced animal performance with higher milk yields recorded for crossbred Awassi × East Friesian (AW × EF) (578 g/day) vs. purebred Awassi (452 g/day) (p < 0.0001). This study provides empirical evidence for the use of rumen-protected and plant-derived oil-infused pellets as supplements under low quality pasture grazing conditions to improve the production performance of purebred Awassi and crossbred AW × EF ewes.

Enhanced Omega-3 Polyunsaturated Fatty Acid Contents in Muscle and Edible Organs of Australian Prime Lambs Grazing Lucerne and Cocksfoot Pastures.

The enhancement of health-beneficial omega-3 long⁻chain (≥C20) polyunsaturated fatty acid (n-3 LC-PUFA) contents in the muscle, liver, heart, and kidney of Australian prime lambs through pasture grazing and supplementation with oil infused pellets was investigated. Forty-eight first-cross prime lambs were randomly assigned into a split-plot design with pasture type as the main plot effect and pellet supplementation as a sub-plot effect in a feeding trial that lasted for nine weeks. The n-3 LC-PUFA content in Longissimus dorsi muscle of all lambs was well above the 30 mg threshold for "omega-3 source" nutrition claim under the Australian Food Standards and Guidelines. Pasture type impacted the fatty acid contents in muscle, heart, and kidney of prime lambs. Lambs grazing cocksfoot grass only had high 18:3n-3 (ALA) and n-3 LC-PUFA contents (67.1 mg/100 g and 55.2 mg/100 g, respectively) in the Longissimus dorsi muscle, which was not significantly different (p > 0.8990) from the contents of lambs grazing only lucerne. Supplementation of pellets with or without oil infusion to grazing lambs generally decreased the ALA and n-3 LC-PUFA contents and increased the n-6/n-3 ratio in the Longissimus dorsi muscle. The fatty acid content in the internal organs of grazing lambs was also affected by pellet supplementation. The liver and kidney of grazing lambs were both "good sources" (60 mg/100 g) of omega-3. The cocksfoot grass showed considerable potential for producing healthy, premium quality meat with high contents of n-3 and n-3 LC-PUFA, which may consequently enhance the omega-3 intake of Australian lamb consumers.

Southern Australian seaweeds: A promising resource for omega-3 fatty acids.

To assess the suitability of southern-Australian macroalgae as potential marine resources for fatty acids (FA), and in particular polyunsaturated fatty acids (PUFA), analysis of 61 species, comprising of 11 Chlorophyta, 17 Phaeophyceae (Ochrophyta) and 33 Rhodophyta, was conducted. Total fatty acid (TFA) concentrations varied considerably (between 0.6 and 7.8 in % of dry weight (DW)) between species, with on average the highest concentrations being in the Phaeophyceae, then the Chlorophyta, and with the Rhodophyta recording the lowest average concentrations. Results revealed significant differences in the fatty acid profiles of the three algal groups. Most species exhibit high proportions of PUFA in their fatty acid profile and a low ratio of n-6/n-3 PUFA. These properties highlight the potential for southern-Australian macroalgae to be used for these FA in food, animal feed and nutraceutical applications.

Supplementation with plant-derived oils rich in omega-3 polyunsaturated fatty acids for lamb production.

In this report, an overview of the health benefits of omega-3 long-chain (≥C20) polyunsaturated fatty acids (n-3 LC-PUFA) and recent progress in using alpha linolenic acid (ALA) rich sources derived from oilseeds to enhance productive performance, n-3 PUFA profiles and sensory properties of lamb for human consumption is reviewed. Omega-3 LC-PUFA can prevent mental health issues and chronic human disorders including cancer, cardiovascular and inflammatory diseases. The median amount of n-3 LC-PUFA consumption is generally lacking in Western diets. More attention is now being paid to the use of innovative nutritional strategies to improve PUFA content in ruminants, which could subsequently increase the content of health-benefitting n-3 LC-PUFA for human consumption. The richest sources of dietary n-3 LC-PUFA are derived from marine products, while forage and oilseeds such as flaxseed, canola, and their oils are abundant in ALA. Numerous studies have shown that dietary ALA increases n-3 LC-PUFA levels of edible tissues. However, other studies concluded that ALA rich supplementation led to no differences in tissue FA profiles because of extensive biohydrogenation of dietary ALA, limited conversion from ALA to n-3 LC-PUFA and low incorporation of n-3 LC-PUFA into edible tissues. Generally, the inclusion of ALA rich sources in lamb diets potentially increases ALA content in lamb. It is proposed that supplementing ruminants with ALA-rich sources at or below 6% can promote n-3 PUFA profiles in lamb and is unlikely to have negative effects on feed intake, growth, carcass and sensory properties.

Omega-3 Long-Chain Fatty Acids in the Heart, Kidney, Liver and Plasma Metabolite Profiles of Australian Prime Lambs Supplemented with Pelleted Canola and Flaxseed Oils.

The objective of the study was to ascertain whether human health beneficial omega-3 long-chain (≥C20) polyunsaturated fatty acid (n-3 LC-PUFA) content in heart, kidney and liver can be enhanced by supplementing prime lambs with graded levels of canola and flaxseed oil. Health status of the lambs, as a consequence of the supplementation, was also investigated by examining their plasma metabolites. Sixty purebred and first-cross lambs were allocated to one of five treatments of lucerne hay basal diet supplemented with isocaloric and isonitrogenous wheat-based pellets without oil inclusion (Control) or graded levels of canola oil at 2.5% (2.5C), 5% (5C), flaxseed oil at 2.5% (2.5F) and 5% (5F) in a completely randomised design. Pre-slaughter blood, post-slaughter kidney, liver and heart samples were analysed for plasma metabolite and fatty acid profiles. Summations of docosapentaenoic acid and docosahexaenoic acid, and total n-3 LC-PUFA were enhanced in the liver and kidney of 5F supplemented lambs with a marked decrease in n-6/n-3 ratio and significant breed differences detected. There were generally no deleterious impacts on animal health status. A combination of 5% oil supplementation and lamb genetics is an effective and strategic management tool for enhancing n-3 LC-PUFA contents of heart, kidney and liver without compromising lamb health.

Australian and New Zealand Fish Oil Products in 2016 Meet Label Omega-3 Claims and Are Not Oxidized.

We provide new fish oil product results to assist industry in Australia and New Zealand and, ultimately, consumers in understanding the high product quality assurance protocols in place, together with the high product quality that has been determined by both industry and independent laboratories. Fish oil capsule products common to Australia and New Zealand were purchased in May 2016 in Richmond, Victoria, Australia. Products were from two groups; five standard fish oil products and five fish oil concentrates. Noting Therapeutic Goods Administration (TGA) requirement for use of standard methods, for all analyses undertaken a laboratory was selected that met the TGA criteria, including with accreditation. Total n-3 content exceeded the label-claimed content for all 10 products, with supplements containing on average 124% of the claimed content (range 115%-136%); eicosapentaenoic acid and docosahexaenoic acid (EPA + DHA) content averaged 109% of the label claim (range 99%-119%). All 10 products (100%) similarly met the international recommended peroxide value (PV) level. Anisidine value (pAV) met the international recommended level for eight of the 10 products, with two products known to contain flavorings that interfere with the pAV test. When accredited laboratories and standard protocols are used, Australian and New Zealand fish oil products have been shown to clearly meet their label claims for EPA + DHA content, and are not oxidized.

Preliminary Validation of a High Docosahexaenoic Acid (DHA) and α-Linolenic Acid (ALA) Dietary Oil Blend: Tissue Fatty Acid Composition and Liver Proteome Response in Atlantic Salmon (Salmo salar) Smolts.

Marine oils are important to human nutrition as the major source of docosahexaenoic acid (DHA), a key omega-3 long-chain (≥C20) polyunsaturated fatty acid (n-3 LC-PUFA) that is low or lacking in terrestrial plant or animal oils. The inclusion of fish oil as main source of n-3 LC-PUFA in aquafeeds is mostly limited by the increasing price and decreasing availability. Fish oil replacement with cheaper terrestrial plant and animal oils has considerably reduced the content of n-3 LC-PUFA in flesh of farmed Atlantic salmon. Novel DHA-enriched oils with high alpha-linolenic acid (ALA) content will be available from transgenic oilseeds plants in the near future as an alternative for dietary fish oil replacement in aquafeeds. As a preliminary validation, we formulated an oil blend (TOFX) with high DHA and ALA content using tuna oil (TO) high in DHA and the flaxseed oil (FX) high in ALA, and assessed its ability to achieve fish oil-like n-3 LC-PUFA tissue composition in Atlantic salmon smolts. We applied proteomics as an exploratory approach to understand the effects of nutritional changes on the fish liver. Comparisons were made between fish fed a fish oil-based diet (FO) and a commercial-like oil blend diet (fish oil + poultry oil, FOPO) over 89 days. Growth and feed efficiency ratio were lower on the TOFX diet. Fish muscle concentration of n-3 LC-PUFA was significantly higher for TOFX than for FOPO fish, but not higher than for FO fish, while retention efficiency of n-3 LC-PUFA was promoted by TOFX relative to FO. Proteomics analysis revealed an oxidative stress response indicative of the main adaptive physiological mechanism in TOFX fish. While specific dietary fatty acid concentrations and balances and antioxidant supplementation may need further attention, the use of an oil with a high content of DHA and ALA can enhance tissue deposition of n-3 LC-PUFA in relation to a commercially used oil blend.

Recent advances in omega-3: Health Benefits, Sources, Products and Bioavailability.

The joint symposium of The Omega-3 Centre and the Australasian Section American Oil Chemists Society; Recent Advances in Omega-3: Health Benefits, Sources, Products and Bioavailability, was held November 7, 2013 in Newcastle, NSW, Australia. Over 115 attendees received new information on a range of health benefits, aquaculture as a sustainable source of supply, and current and potential new and novel sources of these essential omega-3 long-chain (LC, ≥ C20) polyunsaturated fatty acid nutrients (also termed LC omega-3). The theme of "Food versus Fuel" was an inspired way to present a vast array of emerging and ground breaking Omega-3 research that has application across many disciplines. Eleven papers submitted following from the Omega-3 Symposium are published in this Special Issue volume, with topics covered including: an update on the use of the Omega-3 Index (O3I), the effects of dosage and concurrent intake of vitamins/minerals on omega-3 incorporation into red blood cells, the possible use of the O3I as a measure of risk for adiposity, the need for and progress with new land plant sources of docosahexaenoic acid (DHA, 22:6ω3), the current status of farmed Australian and New Zealand fish, and also supplements, in terms of their LC omega-3 and persistent organic pollutants (POP) content, progress with cheap carbon sources in the culture of DHA-producing single cell organisms, a detailed examination of the lipids of the New Zealand Greenshell mussel, and a pilot investigation of the purification of New Zealand hoki liver oil by short path distillation. The selection of papers in this Special Issue collectively highlights a range of forward looking and also new and including positive scientific outcomes occurring in the omega-3 field.

A nutritional-toxicological assessment of Antarctic krill oil versus fish oil dietary supplements.

Fish oil dietary supplements and complementary medicines are pitched to play a role of increasing strategic importance in meeting daily requirements of essential nutrients, such as long-chain (≥ C20, LC) omega-3 polyunsaturated fatty acids and vitamin D. Recently a new product category, derived from Antarctic krill, has been launched on the omega-3 nutriceutical market. Antarctic krill oil is marketed as demonstrating a greater ease of absorption due to higher phospholipid content, as being sourced through sustainable fisheries and being free of toxins and pollutants; however, limited data is available on the latter component. Persistent Organic Pollutants (POP) encompass a range of toxic, man-made contaminants that accumulate preferentially in marine ecosystems and in the lipid reserves of organisms. Extraction and concentration of fish oils therefore represents an inherent nutritional-toxicological conflict. This study aimed to provide the first quantitative comparison of the nutritional (EPA and DHA) versus the toxicological profiles of Antarctic krill oil products, relative to various fish oil categories available on the Australian market. Krill oil products were found to adhere closely to EPA and DHA manufacturer specifications and overall were ranked as containing intermediate levels of POP contaminants when compared to the other products analysed. Monitoring of the pollutant content of fish and krill oil products will become increasingly important with expanding regulatory specifications for chemical thresholds.

DHA-containing oilseed: a timely solution for the sustainability issues surrounding fish oil sources of the health-benefitting long-chain omega-3 oils.

Benefits of long-chain (≥C20) omega-3 oils (LC omega-3 oils) for reduction of the risk of a range of disorders are well documented. The benefits result from eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA); optimal intake levels of these bioactive fatty acids for maintenance of normal health and prevention of diseases have been developed and adopted by national and international health agencies and science bodies. These developments have led to increased consumer demand for LC omega-3 oils and, coupled with increasing global population, will impact on future sustainable supply of fish. Seafood supply from aquaculture has risen over the past decades and it relies on harvest of wild catch fisheries also for its fish oil needs. Alternate sources of LC omega-3 oils are being pursued, including genetically modified soybean rich in shorter-chain stearidonic acid (SDA, 18:4ω3). However, neither oils from traditional oilseeds such as linseed, nor the SDA soybean oil have shown efficient conversion to DHA. A recent breakthrough has seen the demonstration of a land plant-based oil enriched in DHA, and with omega-6 PUFA levels close to that occurring in marine sources of EPA and DHA. We review alternative sources of DHA supply with emphasis on the need for land plant oils containing EPA and DHA.

Readily available sources of long-chain omega-3 oils: is farmed Australian seafood a better source of the good oil than wild-caught seafood?

Seafood consumption enhances intake of omega-3 long-chain (≥C20) polyunsaturated fatty acids (termed LC omega-3 oils). Humans biosynthesize only small amounts of LC-omega-3, so they are considered semi-essential nutrients in our diet. Concern has been raised that farmed fish now contain lower LC omega-3 content than wild-harvested seafood due to the use of oil blending in diets fed to farmed fish. However, we observed that two major Australian farmed finfish species, Atlantic salmon (Salmo salar) and barramundi (Lates calcifer), have higher oil and LC omega-3 content than the same or other species from the wild, and remain an excellent means to achieve substantial intake of LC omega-3 oils. Notwithstanding, LC omega-3 oil content has decreased in these two farmed species, due largely to replacing dietary fish oil with poultry oil. For Atlantic salmon, LC omega-3 content decreased ~30%-50% between 2002 and 2013, and the omega-3/omega-6 ratio also decreased (>5:1 to <1:1). Australian consumers increasingly seek their LC omega-3 from supplements, therefore a range of supplement products were compared. The development and future application of oilseeds containing LC omega-3 oils and their incorporation in aquafeeds would allow these health-benefitting oils to be maximized in farmed Australian seafood. Such advances can assist with preventative health care, fisheries management, aquaculture nutrition, an innovative feed/food industry and ultimately towards improved consumer health.

Characterization of oilseed lipids from "DHA-producing Camelina sativa": a new transformed land plant containing long-chain omega-3 oils.

New and sustainable sources of long-chain (LC, ≥C20) omega-3 oils containing DHA (docosahexaenoic acid, 22:6ω3) are required to meet increasing demands. The lipid content of the oilseed of a novel transgenic, DHA-producing land plant, Camelina sativa, containing microalgal genes able to produce LC omega-3 oils, contained 36% lipid by weight with triacylglycerols (TAG) as the major lipid class in hexane extracts (96% of total lipid). Subsequent chloroform-methanol (CM) extraction recovered further lipid (~50% polar lipid, comprising glycolipids and phospholipids) and residual TAG. The main phospholipid species were phosphatidyl choline and phosphatidyl ethanolamine. The % DHA was: 6.8% (of total fatty acids) in the TAG-rich hexane extract and 4.2% in the polar lipid-rich CM extract. The relative level of ALA (α-linolenic acid, 18:3ω3) in DHA-camelina seed was higher than the control. Major sterols in both DHA- and control camelina seeds were: sitosterol, campesterol, cholesterol, brassicasterol and isofucosterol. C16-C22 fatty alcohols, including iso-branched and odd-chain alcohols were present, including high levels of iso-17:0, 17:0 and 19:0. Other alcohols present were: 16:0, iso-18:0, 18:0 and 18:1 and the proportions varied between the hexane and CM extracts. These iso-branched odd-chain fatty alcohols, to our knowledge, have not been previously reported. These components may be derived from wax esters, or free fatty alcohols.

Polycyclic aromatic hydrocarbon degradation of phytoplankton-associated Arenibacter spp. and description of Arenibacter algicola sp. nov., an aromatic hydrocarbon-degrading bacterium.

Pyrosequencing of the bacterial community associated with a cosmopolitan marine diatom during enrichment with crude oil revealed several Arenibacter phylotypes, of which one (OTU-202) had become significantly enriched by the oil. Since members of the genus Arenibacter have not been previously shown to degrade hydrocarbons, we attempted to isolate a representative strain of this genus in order to directly investigate its hydrocarbon-degrading potential. Based on 16S rRNA sequencing, one isolate (designated strain TG409(T)) exhibited >99% sequence identity to three type strains of this genus. On the basis of phenotypic and genotypic characteristics, strain TG409(T) represents a novel species in the genus Arenibacter, for which the name Arenibacter algicola sp. nov. is proposed. We reveal for the first time that polycyclic aromatic hydrocarbon (PAH) degradation is a shared phenotype among members of this genus, indicating that it could be used as a taxonomic marker for this genus. Kinetic data for PAH mineralization rates showed that naphthalene was preferred to phenanthrene, and its mineralization was significantly enhanced in the presence of glass wool (a surrogate for diatom cell surfaces). During enrichment on hydrocarbons, strain TG409(T) emulsified n-tetradecane and crude oil, and cells were found to be preferentially attached to oil droplets, indicating an ability by the strain to express cell surface amphiphilic substances (biosurfactants or bioemulsifiers) as a possible strategy to increase the bioavailability of hydrocarbons. This work adds to our growing knowledge on the diversity of bacterial genera in the ocean contributing to the degradation of oil contaminants and of hydrocarbon-degrading bacteria found living in association with marine eukaryotic phytoplankton.

Metabolic engineering of biomass for high energy density: oilseed-like triacylglycerol yields from plant leaves.

High biomass crops have recently attracted significant attention as an alternative platform for the renewable production of high energy storage lipids such as triacylglycerol (TAG). While TAG typically accumulates in seeds as storage compounds fuelling subsequent germination, levels in vegetative tissues are generally low. Here, we report the accumulation of more than 15% TAG (17.7% total lipids) by dry weight in Nicotiana tabacum (tobacco) leaves by the co-expression of three genes involved in different aspects of TAG production without severely impacting plant development. These yields far exceed the levels found in wild-type leaf tissue as well as previously reported engineered TAG yields in vegetative tissues of Arabidopsis thaliana and N. tabacum. When translated to a high biomass crop, the current levels would translate to an oil yield per hectare that exceeds those of most cultivated oilseed crops. Confocal fluorescence microscopy and mass spectrometry imaging confirmed the accumulation of TAG within leaf mesophyll cells. In addition, we explored the applicability of several existing oil-processing methods using fresh leaf tissue. Our results demonstrate the technical feasibility of a vegetative plant oil production platform and provide for a step change in the bioenergy landscape, opening new prospects for sustainable food, high energy forage, biofuel and biomaterial applications.

Echium oil is better than rapeseed oil in improving the response of barramundi to a disease challenge.

Pathogen infection stimulates the fatty acid (FA) metabolism and the production of pro-inflammatory derivatives of FA. Barramundi, Lates calcarifer, was fed on a diet rich in preformed long-chain (⩾C20) polyunsaturated fatty acids (LC-PUFA) from fish oil (FO), to compare with diets containing high levels of C18 precursors for LC-PUFA - stearidonic (SDA) and γ-linolenic acid (GLA) - from Echium plantagineum (EO), or rapeseed oil (RO) rich in α-linolenic acid (ALA), but a poor source of LC-PUFA and their precursors. After 6weeks, when growth rates were similar amongst the dietary treatments, a sub-lethal dose of Streptococcus iniae was administered to half of the fish, while the other half were maintained unchallenged and were pair-fed with the infected fish. Under a disease challenge situation, the tissue FA depots depleted at 3days post-infection (DPI) and were then restored to their previous concentrations at 7DPI. During the infection period, EO fish had a higher content of n3 and n6 PUFA in their tissues, higher n3:n6 PUFA ratio and reduced levels of the eicosanoids, TXB2 and 6-keto-PGF1α, in their plasma compared with RO fish. Fish fed on FO and EO had a longer lasting and enduring response in their FA and eicosanoid concentrations, following a week of bacterial infection, compared with those fed on RO. EO, containing SDA and GLA and with a comparatively higher n3:n6 PUFA ratio, proved more effective than RO in compensating for immunity stress.

Coping with sub-optimal water temperature: modifications in fatty acid profile of barramundi as influenced by dietary lipid.

Metabolic responses to sub-optimal temperature deplete lipid depots, remodel membrane lipid and alter the fatty acid profile in the whole body and tissues of ectothermic vertebrates including fish. The magnitude of these changes may depend on dietary history including oil sources with different fatty acid compositions. Barramundi, Lates calcarifer (Perciformes, Latidae), a tropical ectothermic fish, was fed on diets either rich in dietary long-chain (≥C(20)) polyunsaturated fatty acids (LC-PUFA) from fish oil, rich in stearidonic and γ-linolenic acid (SDA and GLA, respectively) from Echium plantagineum, or rapeseed oil deficient in LC-PUFA. Following 5 weeks at the optimum temperature of 30 °C when growth rates were comparable amongst dietary treatments, water temperature was dropped to 20 °C for 1 week for half of the animals and maintained at 30 °C for the other half. Decreased temperature increased the liver and skeletal muscle content of LC-PUFA in fish fed on echium oil compared with rapeseed oil, while dietary LC-PUFA depots in fish oil fed-fish depleted rapidly in the week of sub-optimal temperature. The lipid unsaturation index of cellular membrane in the liver and muscle increased under low temperature at the same rate regardless of dietary oil. Therefore, rapid exposure of an ectothermic vertebrate to a lower and sub-optimal temperature caused significant modulation in fatty acid composition. We propose that the tolerance of barramundi, a representative of tropical farmed fish, to sub-optimal temperature will be enhanced when fatty acid substrates closer to the LC-PUFA are available in their diet.