Docosahexaenoic Acid Is Naturally Concentrated at the sn-2 Position in Triacylglycerols of the Australian Thraustochytrid Aurantiochytrium sp. Strain TC 20.

The Labyrinthulomycetes or Labyrinthulea are a class of protists that produce a network of filaments that enable the cells to glide along and absorb nutrients. One of the main two Labyrinthulea groups is the thraustochytrids, which are becoming an increasingly recognised and commercially used alternate source of long-chain (LC, ≥C20) omega-3 containing oils. This study demonstrates, to our knowledge for the first time, the regiospecificity of the triacylglycerol (TAG) fraction derived from Australian thraustochytrid Aurantiochytrium sp. strain TC 20 obtained using 13C nuclear magnetic resonance spectroscopy (13C NMR) analysis. The DHA present in the TC 20 TAG fraction was determined to be concentrated in the sn-2 position, with TAG (16:0/22:6/16:0) identified as the main species present. The sn-2 preference is similar to that found in salmon and tuna oil, and differs to seal oil containing largely sn-1,3 LC-PUFA. A higher concentration of sn-2 DHA occurred in the thraustochytrid TC 20 oil compared to that of tuna oil.

Adjustments in fatty acid composition is a mechanism that can explain resilience to marine heatwaves and future ocean conditions in the habitat-forming seaweed Phyllospora comosa (Labillardière) C.Agardh.

Marine heatwaves are extreme events that can have profound and lasting impacts on marine species. Field observations have shown seaweeds to be highly susceptible to marine heatwaves, but the physiological drivers of this susceptibility are poorly understood. Furthermore, the effects of marine heatwaves in conjunction with ocean warming and acidification are yet to be investigated. To address this knowledge gap, we conducted a laboratory culture experiment in which we tested the growth and physiological responses of Phyllospora comosa juveniles from the southern extent of its range (43-31°S) to marine heatwaves, ocean warming and acidification. We used a 'collapsed factorial design' in which marine heatwaves were superimposed on current (today's pH and temperature) and future (pH and temperature projected by 2100) ocean conditions. Responses were tested both during the heatwaves, and after a 7-day recovery period. Heatwaves reduced net photosynthetic rates in both current and future conditions, while respiration rates were elevated under heatwaves in the current conditions only. Following the recovery period, there was little evidence of heatwaves having lasting negative effects on growth, photosynthesis or respiration. Exposure to heatwaves, future ocean conditions or both caused an increase in the degree of saturation of fatty acids. This adjustment may have counteracted negative effects of elevated temperatures by decreasing membrane fluidity, which increases at higher temperatures. Furthermore, P. comosa appeared to down-regulate the energetically expensive carbon dioxide concentrating mechanism in the future conditions with a reduction in δ13 C values detected in these treatments. Any saved energy arising from this down-regulation was not invested in growth and was likely invested in the adjustment of fatty acid composition. This adjustment is a mechanism by which P. comosa and other seaweeds may tolerate the negative effects of ocean warming and marine heatwaves through benefits arising from ocean acidification.

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.

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.

Liver proteome response of pre-harvest Atlantic salmon following exposure to elevated temperature.

BACKGROUND: Atlantic salmon production in Tasmania (Southern Australia) occurs near the upper limits of the species thermal tolerance. Summer water temperatures can average over 19 °C over several weeks and have negative effects on performance and health. Liver tissue exerts important metabolic functions in thermal adaptation. With the aim of identifying mechanisms underlying liver plasticity in response to chronic elevated temperature in Atlantic salmon, label-free shotgun proteomics was used to explore quantitative protein changes after 43 days of exposure to elevated temperature. RESULTS: A total of 276 proteins were differentially (adjusted p-value < 0.05) expressed between the control (15 °C) and elevated (21 °C) temperature treatments. As identified by Ingenuity Pathway Analysis (IPA), transcription and translation mechanisms, protein degradation via the proteasome, and cytoskeletal components were down-regulated at elevated temperature. In contrast, an up-regulated response was identified for NRF2-mediated oxidative stress, endoplasmic reticulum stress, and amino acid degradation. The proteome response was paralleled by reduced fish condition factor and hepato-somatic index at elevated temperature. CONCLUSIONS: The present study provides new evidence of the interplay among different cellular machineries in a scenario of heat-induced energy deficit and oxidative stress, and refines present understanding of how Atlantic salmon cope with chronic exposure to temperature near the upper limits of thermal tolerance.

Sequential protein extraction as an efficient method for improved proteome coverage in larvae of Atlantic salmon (Salmo salar).

Understanding diet- and environmentally induced physiological changes in fish larvae is a major goal for the aquaculture industry. Proteomic analysis of whole fish larvae comprising multiple tissues offers considerable potential but is challenging due to the very large dynamic range of protein abundance. To extend the coverage of the larval phase of the Atlantic salmon (Salmo salar) proteome, we applied a two-step sequential extraction (SE) method, based on differential protein solubility, using a nondenaturing buffer containing 150 mM NaCl followed by a denaturing buffer containing 7 M urea and 2 M thiourea. Extracts prepared using SE and one-step direct extraction were characterized via label-free shotgun proteomics using nanoLC-MS/MS (LTQ-Orbitrap). SE partitioned the proteins into two fractions of approximately equal amounts, but with very distinct protein composition, leading to identification of ∼40% more proteins than direct extraction. This fractionation strategy enabled the most detailed characterization of the salmon larval proteome to date and provides a platform for greater understanding of physiological changes in whole fish larvae. The MS data are available via the ProteomeXchange Consortium PRIDE partner repository, dataset PXD003366.

Direct determination of fatty acids in fish tissues: quantifying top predator trophic connections.

Fatty acids are a valuable tool in ecological studies because of the large number of unique structures synthesized. They provide versatile signatures that are being increasingly employed to delineate the transfer of dietary material through marine and terrestrial food webs. The standard procedure for determining fatty acids generally involves lipid extraction followed by methanolysis to produce methyl esters for analysis by gas chromatography. By directly transmethylating ~50 mg wet samples and adding an internal standard it was possible to greatly simplify the analytical methodology to enable rapid throughput of 20-40 fish tissue fatty acid analyses a day including instrumental analysis. This method was verified against the more traditional lipid methods using albacore tuna and great white shark muscle and liver samples, and it was shown to provide an estimate of sample dry mass, total lipid content, and a condition index. When large fatty acid data sets are generated in this way, multidimensional scaling, analysis of similarities, and similarity of percentages analysis can be used to define trophic connections among samples and to quantify them. These routines were used on albacore and skipjack tuna fatty acid data obtained by direct methylation coupled with literature values for krill. There were clear differences in fatty acid profiles among the species as well as spatial differences among albacore tuna sampled from different locations.

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.

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.

Commentary on a trial comparing krill oil versus fish oil.

Considerable interest exists presently in comparing the performance of krill oil (KO) and fish oil (FO) supplements. Ramprasath et al. (Lipids Health Dis12:178, 2013) have recently compared use of KO and FO in a trial with healthy individuals to examine which oil is more effective in increasing n-3 PUFA, decreasing the n-6:n-3 ratio and improving the omega-3 index. The authors concluded that KO was more effective than FO for all three criteria. However, careful examination of the fatty acid profiles of the oils used showed that the FO used was not a typical FO; it contained linoleic acid as the dominant fatty acid (32%) and an n-6:n-3 ratio of >1. Due to the fatty acid profile being non-representative of typically commercially marketed FO, the conclusions presented by Ramrasath et al. (Lipids Health Dis12:178, 2013) are not justified and misleading. Considerable care is needed in ensuring that such comparative trials do not use inappropriate ingredients.

No distinct local cuisines among humpback whales: A population diet comparison in the Southern Hemisphere.

Southern hemisphere humpback whale (Megaptera novaeangliae, SHHW) breeding populations follow a high-fidelity Antarctic krill (Euphausia superba) diet while feeding in distinct sectors of the Southern Ocean. Their capital breeding life history requires predictable ecosystem productivity to fuel migration and migration-related behaviours. It is therefore postulated that populations feeding in areas subject to the strongest climate change impacts are more likely to show the first signs of a departure from a high-fidelity krill diet. We tested this hypothesis by investigating blubber fatty acid profiles and skin stable isotopes obtained from five SHHW populations in 2019, and comparing them to Antarctic krill stable isotopes sampled in three SHHW feeding areas in the Southern Ocean in 2019. Fatty acid profiles and δ13C and δ15N varied significantly among all five populations, however, calculated trophic positions did not (2.7 to 3.1). Similarly, fatty acid ratios, 16:1ω7c/16:0 and 20:5ω3/22:6ω3 were above 1, showing that whales from all five populations are secondary heterotrophs following an omnivorous diet with a diatom-origin. Thus, evidence for a potential departure from a high-fidelity Antarctic krill diet was not seen in any population. δ13C of all populations were similar to δ13C of krill sampled in productive upwelling areas or the marginal sea-ice zone. Consistency in trophic position and diet origin but significant fatty acid and stable isotope differences demonstrate that the observed variability arises at lower trophic levels. Our results indicate that, at present, there is no evidence of a divergence from a high-fidelity krill diet. Nevertheless, the characteristic isotopic signal of whales feeding in productive upwelling areas, or in the marginal sea-ice zone, implies that future cryosphere reductions could impact their feeding ecology.

Polycyclovorans algicola gen. nov., sp. nov., an aromatic-hydrocarbon-degrading marine bacterium found associated with laboratory cultures of marine phytoplankton.

A strictly aerobic, halotolerant, rod-shaped bacterium, designated strain TG408, was isolated from a laboratory culture of the marine diatom Skeletonema costatum (CCAP1077/1C) by enrichment with polycyclic aromatic hydrocarbons (PAHs) as the sole carbon source. 16S rRNA gene sequence analysis placed this organism within the order Xanthomonadales of the class Gammaproteobacteria. Its closest relatives included representatives of the Hydrocarboniphaga-Nevskia-Sinobacter clade (<92% sequence similarity) in the family Sinobacteraceae. The strain exhibited a narrow nutritional spectrum, preferring to utilize aliphatic and aromatic hydrocarbon compounds and small organic acids. Notably, it displayed versatility in degrading two- and three-ring PAHs. Moreover, catechol 2,3-dioxygenase activity was detected in lysates, indicating that this strain utilizes the meta-cleavage pathway for aromatic compound degradation. Cells produced surface blebs and contained a single polar flagellum. The predominant isoprenoid quinone of strain TG408 was Q-8, and the dominant fatty acids were C(16:0), C(16:1) ω7c, and C(18:1) ω7c. The G+C content of the isolate's DNA was 64.3 mol% ± 0.34 mol%. On the basis of distinct phenotypic and genotypic characteristics, strain TG408 represents a novel genus and species in the class Gammaproteobacteria for which the name Polycyclovorans algicola gen. nov., sp. nov., is proposed. Quantitative PCR primers targeting the 16S rRNA gene of this strain were developed and used to show that this organism is found associated with other species of marine phytoplankton. Phytoplankton may be a natural biotope in the ocean where new species of hydrocarbon-degrading bacteria await discovery and which contribute significantly to natural remediation processes.

High cell density cultivation of a novel Aurantiochytrium sp. strain TC 20 in a fed-batch system using glycerol to produce feedstock for biodiesel and omega-3 oils.

A recently isolated Australian Aurantiochytrium sp. strain TC 20 was investigated using small-scale (2 L) bioreactors for the potential of co-producing biodiesel and high-value omega-3 long-chain polyunsaturated fatty acids. Higher initial glucose concentration (100 g/L compared to 40 g/L) did not result in markedly different biomass (48 g/L) or fatty acid (12-14 g/L) yields by 69 h. This comparison suggests factors other than carbon source were limiting biomass production. The effect of both glucose and glycerol as carbon sources for Aurantiochytrium sp. strain TC 20 was evaluated in a fed-batch process. Both glucose and glycerol resulted in similar biomass yields (57 and 56 g/L, respectively) by 69 h. The agro-industrial waste from biodiesel production-glycerol-is a suitable carbon source for Aurantiochytrium sp. strain TC 20. Approximately half the fatty acids from Aurantiochytrium sp. strain TC 20 are suitable for development of sustainable, low emission sources of transportation fuels and bioproducts. To further improve biomass and oil production, fortification of the feed with additional nutrients (nitrogen sources, trace metals and vitamins) improved the biomass yield from 56 g/L (34 % total fatty acids) to 71 g/L (52 % total fatty acids, cell dry weight) at 69 h; these yields are to our knowledge around 70 % of the biomass yields achieved, however, in less than half of the time by other researchers using glycerol and markedly greater than achieved using other industrial wastes. The fast growth and suitable fatty acid profile of this newly isolated Aurantiochytrium sp. strain TC 20 highlights the potential of co-producing the drop-in biodiesel and high value omega-3 oils.

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.

Fatty acid profiles of more than 470 marine species from the Southern Hemisphere.

Lipid and fatty acid datasets are commonly used to assess the nutritional composition of organisms, trophic ecology, and ecosystem dynamics. Lipids and their fatty acid constituents are essential nutrients to all forms of life because they contribute to biological processes such as energy flow and metabolism. Assessment of total lipids in tissues of organisms provides information on energy allocation and life-history strategies and can be an indicator of nutritional condition. The analysis of an organism's fatty acids is a widely used technique for assessing nutrient and energy transfer, and dietary interactions in food webs. Although there have been many published regional studies that assessed lipid and fatty acid compositions, many only report the mean values of the most abundant fatty acids. There are limited individual records available for wider use in intercomparison or macro-scale studies. This dataset consists of 4856 records of individual and pooled samples of at least 470 different marine consumer species sampled from tropical, temperate, and polar regions around Australia and in the Southern, Indian, and Pacific Oceans from 1989 to 2018. This includes data for a diverse range of taxa (zooplankton, fish, cephalopods, chondrichthyans, and marine mammals), size ranges (0.02 cm to ~13 m), and that cover a broad range of trophic positions (2.0-4.6). When known, we provide a record of species name, date of sampling, sampling location, body size, relative (%) measurements of tissue-specific total lipid content and abundant fatty acids, and absolute content (mg 100 g-1 tissue) of eicosapentaenoic acid (EPA, 20:5n3) and docosahexaenoic acid (DHA, 22:6n3) as important long-chain (≥C20 ) polyunsaturated omega-3 fatty acids. These records form a solid basis for comparative studies that will facilitate a broad understanding of the spatial and temporal distribution of marine lipids globally. The dataset also provides reference data for future dietary assessments of marine predators and model assessments of potential impacts of climate change on the availability of marine lipids and fatty acids. There are 480 data records within our data file for which the providers have requested that permission for reuse be granted, with the likely condition that they are included as a coauthor on the reporting of the dataset. Records with this condition are indicated by a "yes" under "Conditions_of_data_use" in Data S1: Marineconsumer_FAdata.csv (see Table 2 in Metadata S1 for more details). For all other data records marked as "No" under "Conditions_of_data_use," there are no copyright restrictions for research and/or teaching purposes. We request that users acknowledge use of the data in publications, research proposals, websites, and other outlets via formal citation of this work and original data sources as applicable.

Porticoccus hydrocarbonoclasticus sp. nov., an aromatic hydrocarbon-degrading bacterium identified in laboratory cultures of marine phytoplankton.

A marine bacterium, designated strain MCTG13d, was isolated from a laboratory culture of the dinoflagellate Lingulodinium polyedrum CCAP1121/2 by enrichment with polycyclic aromatic hydrocarbons (PAHs) as the sole carbon source. Based on 16S rRNA gene sequence comparisons, the strain was most closely related to Porticoccus litoralis IMCC2115(T) (96.5%) and to members of the genera Microbulbifer (91.4 to 93.7%) and Marinimicrobium (90.4 to 92.0%). Phylogenetic trees showed that the strain clustered in a distinct phyletic line in the class Gammaproteobacteria for which P. litoralis is presently the sole cultured representative. The strain was strictly aerobic, rod shaped, Gram negative, and halophilic. Notably, it was able to utilize hydrocarbons as sole sources of carbon and energy, whereas sugars did not serve as growth substrates. The predominant isoprenoid quinone of strain MCTG13d was Q-8, and the dominant fatty acids were C(16:1ω7c), C(18:1ω7c), and C(16:0). DNA G+C content for the isolate was 54.9 ± 0.42 mol%. Quantitative PCR primers targeting the 16S rRNA gene of this strain showed that this organism was common in other laboratory cultures of marine phytoplankton. On the basis of phenotypic and genotypic characteristics, strain MCTG13d represents a novel species of Porticoccus, for which the name Porticoccus hydrocarbonoclasticus sp. nov. is proposed. The discovery of this highly specialized hydrocarbon-degrading bacterium living in association with marine phytoplankton suggests that phytoplankton represent a previously unrecognized biotope of novel bacterial taxa that degrade hydrocarbons in the ocean.

Transgenic production of arachidonic acid in oilseeds.

We describe a transgenic microalgal Δ9-elongase pathway transformed in both Brassica napus and Arabidopsis thaliana seed resulting in the production of arachidonic acid (ARA). This pathway is noteworthy for both the production of ARA in seed tissue and the low levels of intermediate C20 fatty acids that accumulate. We also demonstrate that the arachidonic acid is naturally enriched at the sn2 position in triacylglycerol. This is the first report of ARA production by the Δ9-elongase pathway in an oilseed.

Jumping on the omega-3 bandwagon: distinguishing the role of long-chain and short-chain omega-3 fatty acids.

Omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) are almost unanimously recognized for their health benefits, while only limited evidence of any health benefit is currently available specifically for the main precursor of these fatty acids, namely α-linolenic acid (ALA, 18:3n-3). However, both the n-3 LC-PUFA and the short-chain C18 PUFA (i.e., ALA) are commonly referred to as "omega-3" fatty acids, and it is difficult for consumers to recognize this difference. A current gap of many food labelling legislations worldwide allow products containing only ALA and without n-3 LC-PUFA to be marketed as "omega-3 source" and this misleading information can negatively impact the ability of consumers to choose more healthy diets. Within the context of the documented nutritional and health promoting roles of omega-3 fatty acids, we briefly review the different metabolic fates of dietary ALA and n-3 LC-PUFA. We also review food sources rich in n-3 LC-PUFA, some characteristics of LC-PUFA and current industry and regulatory trends. A further objective is to present a case for regulatory bodies to clearly distinguish food products containing only ALA from foods containing n-3 LC-PUFA. Such information, when available, would then avoid misleading information and empower consumers to make a more informed choice in their food purchasing behavior.

Algiphilus aromaticivorans gen. nov., sp. nov., an aromatic hydrocarbon-degrading bacterium isolated from a culture of the marine dinoflagellate Lingulodinium polyedrum, and proposal of Algiphilaceae fam. nov.

A strictly aerobic, halotolerant, Gram-stain-negative, rod-shaped bacterium, designated strain DG1253(T), was isolated from a laboratory culture of the marine dinoflagellate Lingulodinium polyedrum (CCAP 1121/2). The strain was able to degrade two- and three-ring polycyclic aromatic hydrocarbons. It exhibited a narrow nutritional spectrum, preferring to utilize aliphatic and aromatic hydrocarbon compounds and small organic acids. Cells produced surface blebs and contained a single polar flagellum. The predominant isoprenoid quinone of strain DG1253(T) was Q-8. The fatty acid profile was dominated by C(18:1)ω7c. The mean DNA G+C content of strain DG1253(T) was 63.6 ± 0.25 mol%. 16S rRNA gene sequence analysis placed this organism within the order Xanthomonadales of the class Gammaproteobacteria. Its closest relatives included representatives of the Hydrocarboniphaga-Nevskia-Sinobacter clade (≤ 89.9% 16S rRNA gene sequence similarity) in the family Sinobacteraceae. On the basis of distinct phenotypic and genotypic characteristics, strain DG1253(T) is considered to represent a novel species of a new genus in the class Gammaproteobacteria, for which the name Algiphilus aromaticivorans gen. nov., sp. nov. is proposed. The type strain of the type species, Algiphilus aromaticivorans, is DG1253(T) (=ATCC BAA-2243(T)=DSM 24793(T)). In addition, a new family, Algiphilaceae fam. nov., is proposed to accommodate the genus Algiphilus.

Biodiscovery of new Australian thraustochytrids for production of biodiesel and long-chain omega-3 oils.

Heterotrophic growth of thraustochytrids has potential in co-producing a feedstock for biodiesel and long-chain (LC, ≥C(20)) omega-3 oils. Biodiscovery of thraustochytrids from Tasmania (temperate) and Queensland (tropical), Australia, covered a biogeographic range of habitats including fresh, brackish, and marine waters. A total of 36 thraustochytrid strains were isolated and separated into eight chemotaxonomic groups (A-H) based on fatty acid (FA) and sterol composition which clustered closely with four different genera obtained by 18S rDNA molecular identification. Differences in the relative proportions (%FA) of long-chain C(20), C(22), omega-3, and omega-6 polyunsaturated fatty acids (PUFA), including docosahexaenoic acid (DHA), docosapentaenoic acid, arachidonic acid, eicosapentaenoic acid (EPA), and saturated FA, as well as the presence of odd-chain PUFA (OC-PUFA) were the major factors influencing the separation of these groups. OC-PUFA were detected in temperate strains of groups A, B, and C (Schizochytrium and Thraustochytrium). Group D (Ulkenia) had high omega-3 LC-PUFA (53% total fatty acids (TFA)) and EPA up to 11.2% TFA. Strains from groups E and F (Aurantiochytrium) contained DHA levels of 50-61% TFA after 7 days of growth in basal medium at 20 °C. Groups G and H (Aurantiochytrium) strains had high levels of 15:0 (20-30% TFA) and the sum of saturated FA was in the range of 32-51%. ß,ß-Carotene, canthaxanthin, and astaxanthin were identified in selected strains. Phylogenetic and chemotaxonomic groupings demonstrated similar patterns for the majority of strains. Our results demonstrate the potential of these new Australian thraustochytrids for the production of biodiesel in addition to omega-3 LC-PUFA-rich oils.