Experimental reduction in dietary omega-3 polyunsaturated fatty acids depresses sperm competitiveness

 The health benefits of diets containing rich sources of long-chain omega-3 polyunsaturated fatty acids (n-3 LC-PUFA) are well documented and include reductions in the risk of several diseases typical of Western societies. The dietary intake of n-3 LC-PUFA has also been linked to fertility, and there is abundant evidence that a range of ejaculate traits linked to fertility in humans, livestock and other animals depend on an adequate intake of n-3 LC-PUFA from dietary sources. However, relatively few studies have explored how n-3 LC-PUFA influence reproductive fitness, particularly in the context of sexual selection. Here, we show that experimental reduction in the level of n-3 LC-PUFA in the diet of guppies (Poecilia reticulata) depresses a male’s share of paternity when sperm compete for fertilization, confirming that the currently observed trend for reduced n-3 LC-PUFA in western diets has important implications for individual reproductive fitness.

Solvent-induced 7R-dioxygenase activity of soybean 15-lipoxygenase-1 in the formation of omega-3 DPA-derived resolvin analogs

The resolvin family contains important anti-inflammatory and pro-resolution compounds enzymatically derived in vivo from the polyunsaturated omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). More recently, docosapentaenoic acid (DPA) has emerged as another potentially important precursor in the biological production of resolvin compounds. In this work we have used medium engineering to develop a simple method for the controlled synthesis of two di-hydroxylated diastereomers of DPAn-3 catalyzed by soybean 15-lipoxygenase-1 (15-sLOX-1) in the presence of short chain n-alcohols, including methanol, ethanol and propan-1-ol. The complete structures of the two major products, 7S,17S-dihydroxydocosapenta-8Z,10E,13Z,15E,19Z-enoic acid (7S,17S-diHDPAn-3) and 7R,17S-dihydroxydocosapenta-8Z,10E,13Z,15E,19Z- enoic acid (7R,17S-diHDPAn-3), have been elucidated using spectroscopic analysis. The alcohol-dependent R-dioxygenase activity of soybean 15-lipoxygenase with mono-hydroperoxide intermediate substrates has also been demonstrated with other biologically relevant PUFAs, including DHA, EPA and ARA. The developed method has applications in the production of closely related isomers of naturally occurring resolvins and protectins, demonstrating the versatility of 15-sLOX-1 as a biocatalyst. © 2014 Elsevier B.V.

Omega-3 fatty acid production from enzyme saccharified hemp hydrolysate using a novel marine thraustochytrid strain

In this work, a newly isolated marine thraustochytrid strain, Schizochytrium sp. DT3, was used for omega-3 fatty acid production by growing on lignocellulose biomass obtained from local hemp hurd (Cannabis sativa) biomass. Prior to enzymatic hydrolysis, hemp was pretreated with sodium hydroxide to open the biomass structure for the production of sugar hydrolysate. The thraustochytrid strain was able to grow on the sugar hydrolysate and accumulated polyunsaturated fatty acids (PUFAs). At the lowest carbon concentration of 2%, the PUFAs productivity was 71% in glucose and 59% in the sugars hydrolysate, as a percentage of total fatty acids. Saturated fatty acids (SFAs) levels were highest at about 49% of TFA using 6% glucose as the carbon source. SFAs of 41% were produced using 2% of SH. This study demonstrates that SH produced from lignocellulose biomass is a potentially useful carbon source for the production of omega-3 fatty acids in thraustochytrids, as demonstrated using the new strain, Schizochytrium sp. DT3.

Propyl gallate and butylated hydroxytoluene influence the accumulation of saturated fatty acids, omega-3 fatty acid and carotenoids in thraustochytrids

Schizochytrium sp. S31 was shown to have potential for production of the functional food ingredients docosahexaenoic acid (DHA) and astaxanthin, with coproduction of biodiesel. Biomass and lipid levels were greater with glycerol than with glucose as carbon source. Addition of propyl gallate or butylated hydroxytoluene to the media resulted in increased biomass and lipid levels, with propyl gallate being the more effective of the two antioxidants. Medium supplementation with propyl gallate at 0.03% and glycerol as the carbon source resulted in enhanced biomass productivity (28.50 g L^-1), lipid accumulation (24.87 g L^-1) and astaxanthin levels (452.26 μg L^-1).

Dietary sources, current intakes, and nutritional role of omega-3 docosapentaenoic acid

Fish oils and long-chain omega-3 fatty acids are well recognized for their critical role in human diets. Docosapentaenoic acid (DPA, 22:5n-3) has always been a part of healthy nutrition, since infants obtain almost as much DPA as DHA from human milk. Fish oil supplements and ingredients, oily fish, and grass-fed beef can serve as the primary DPA sources for the general population. Although the DPA levels in fish oils are substantially lower than those of EPA and DHA, concentrated DPA products are now becoming commercially available, and DPA-based drugs are under development. Epidemiological studies show that similar to eicosapentaenoic (EPA, 20:5n-3) and docosahexaenoic (DHA, 22:6n-3) acids, DPA is linked to various improvements in human health, perhaps owing to its structural similarity to the other two molecules. Studies in mammals, platelets, and cell cultures have demonstrated that DPA reduces platelet aggregation, and improves lipid metabolism, endothelial cell migration, and resolution of chronic inflammation. Further, other in vivo and in vitro studies have shown that DPA can improve neural health. A human supplementation trial with 99.8% pure DPA suggested that it serves as a storage depot for EPA and DHA in the human body. Future randomized controlled human trials with purified DPA will help clarify its effects on human health. They may confirm the available evidence pointing to its nutritional and biological functions, unique or overlapping with those of EPA and DHA.

What is the most effective way of increasing the bioavailability of dietary long chain omega-3 fatty acids-daily vs. weekly administration of fish oil?

The recommendations on the intake of long chain omega-3 polyunsaturated fatty acids (n-3 LC-PUFA) vary from eating oily fish ("once to twice per week") to consuming specified daily amounts of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) ("250-500 mg per day"). It is not known if there is a difference in the uptake/bioavailability between regular daily consumption of supplementsvs. consuming fish once or twice per week. In this study, the bioavailability of a daily dose of n-3 LC-PUFA (Constant treatment), representing supplements, vs. a large weekly dose of n-3 LC-PUFA (Spike treatment), representing consuming once or twice per week, was assessed. Six-week old healthy male Sprague-Dawley rats were fed either a Constant treatment, a Spike treatment or Control treatment (no n-3 LC-PUFA), for six weeks. The whole body, tissues and faeces were analysed for fatty acid content. The results showed that the major metabolic fate of the n-3 LC-PUFA (EPA+docosapentaenoic acid (DPA) + DHA) was towards catabolism (β-oxidation) accounting for over 70% of total dietary intake, whereas deposition accounted less than 25% of total dietary intake. It was found that significantly more n-3 LC-PUFA were β-oxidised when originating from the Constant treatment (84% of dose), compared with the Spike treatment (75% of dose). Conversely, it was found that significantly more n-3 LC-PUFA were deposited when originating from the Spike treatment (23% of dose), than from the Constant treatment (15% of dose). These unexpected findings show that a large dose of n-3 LC-PUFA once per week is more effective in increasing whole body n-3 LC-PUFA content in rats compared with a smaller dose delivered daily.

Effects of omega-3 PUFA on the vitamin E and glutathione antioxidant defense system in individuals at ultra-high risk of psychosis

BACKGROUND: Oxidative stress and impaired antioxidant defenses are reported in schizophrenia and are associated with disturbed neurodevelopment, brain structural alterations, glutamatergic imbalance, increased negative symptoms, and cognitive impairment. There is evidence that oxidative stress predates the onset of acute psychotic illness. Here, we investigate the effects of omega-3 PUFA on the vitamin E and glutathione antioxidant defense system (AODS). METHOD: In 64 help-seeking UHR-individuals (13-25 years of age), vitamin E levels and glutathione were investigated before and after 12 weeks of treatment with either 1.2g/d omega-3 (PUFA-E) or saturated fatty acids (SFA-E), with each condition also containing 30.4mg/d alpha-tocopherol to ensure absorption without additional oxidative risk. RESULTS: In multivariate tests, the effects on the AODS (alpha-tocopherol, total glutathione) were not significantly different (p=0.13, p=0.11, respectively) between treatment conditions. According to univariate findings, only PUFA-E caused a significant alpha-tocopherol increase, while PUFA-E and SFA-E caused a significant gamma- and delta-tocopherol decrease. Total glutathione (GSHt) was decreased by PUFA-E supplementation. CONCLUSION: Effects of the PUFA-E condition on the vitamin E and glutathione AODS could be mechanisms underlying its clinical effectiveness. In terms of the vitamin E protection system, PUFA-E seems to directly support the antioxidative defense at membrane level. The effect of PUFA-E on GSHt is not yet fully understood, but could reflect antioxidative effects, resulting in decreased demand for glutathione. It is still necessary to further clarify which type of PUFA/antioxidant combination, and in which dose, is effective at each stage of psychotic illness.

Synthetic biology based metabolic engineering of saccharomyces cerevisiae for omega 6 and 3 PUFA production

Metabolic engineering of PUFA biosynthesis pathway using codon optimized DGA1 (Diacylglycerol acyltransferase), FAA3 (Acyl-CoA synthetase), desaturase genes named D9D, D12D, D5D, D6D, D17D and D6E elongase gene was studied in S. cerevisiae. Engineered yeast strains successfully demonstrated increase in lipid accumulation, and heterologous biosynthesis of linoleic, γ-linolenic, dihomo γ-linolenic, arachidonic and eicosapentaenoic acid.

Fermentation of omega-3 and carotenoid producing marine microorganisms

 The objective of the thesis was to understand and develop the process of fermentation in marine microorganisms for the production of omega-3 fatty acids and carotenoids. Among marine microorganisms, thraustochytrids that belong to phytoplankton group was identified as sources of omega-3 fatty acids and other valuable co-products. In this research, more efficient and cost-effective production of omega-3 oils and other value added products was discussed by addressing the below key objectives. Fermentation strategy using lower cost raw materials, particularly carbon. Screening for new strains that can naturally produce high levels of PUFAs together with useful co-products that can be harvested along with the omega-3 oil.

Omega-3 fatty acids and zinc in neuronal cell homeostasis and survival

 The main focus of this research was to investigate possible link between zinc, DHA, apoptosis and cell survival. Study also analyses the benefits of omega-3 FA’s and the link between free zinc availability to neurodegeneration. Furthermore, this study was focused on developing a suitable cell culture model for neuronal research.

Bioavailability of long chain omega-3 polyunsaturated fatty acids

It has been well documented that the ingestion of omega-3 polyunsaturated fatty acids (n-3 LC-PUFA) can improve health. However, little scientific attention has been paid on the actual bioavailability of n-3 LC-PUFA. In this thesis, several nutritional questions related to bioavailability of n-3 LC-PUFA have been answered.

Recovery of omega-3 profiles of cultivated abalone by dietary macroalgae supplementation

Formulated feeds for cultivated abalone have received considerable attention to improve the feeding costs, efficiency and productivity of the growing cultivated abalone industry in Australia. Less attention has been given to the resulting quality of the seafood product from formulated feeds and the effect of the reduction of marine ingredients in these feeds. Marine n-3 fatty acids are particularly important nutritional factors of seafood that are lacking from feed ingredients at the base of the aquaculture production chain. Considering that this important category of nutrients is already deficient in the western diet, further erosion through cultivated seafood products is of concern. This study tested the effect of three diet categories on the fatty acid profiles of commercially farmed abalone. Two commercial formulated feeds, Ulva spp. macroalgae, and combinations of the formulated feeds and macroalgae were fed to six replicate baskets of 20 abalone (~62 mm) for 12 months in Victoria, Australia. The macroalgae diets represented overall lower fatty acid content, including eicosapentaenoic (EPA) and docosahexaenoic (DHA), but higher relative amounts of alpha linolenic acid (ALA) and docosapentaenoic acid (DPA), than the formulated feeds. The total lipid content of abalone tissue did not vary substantially across diet categories; however, the ratio of n-6:n-3 fatty acids decreased incrementally with the increasing macroalgae content in the diet and by a factor of two with a 100 % macroalgae diet. Also, equal or higher content of all important long chain n-3 fatty acids was achieved with a macroalgae diet despite the lower dietary content of some of these fatty acids. Marine fatty acid-deplete aquaculture feeds can result in decreased n-3 fatty acid content in the abalone tissue. The inclusion of macroalgae in the diet of abalone can offset this reduction.

A computational search for lipases that can preferentially hydrolyze long-chain omega-3 fatty acids from fish oil triacylglycerols

Consumption of long-chain omega-3 fatty acids is known to decrease the risk of major cardiovascular events. Lipases, a class of triacylglycerol hydrolases, have been extensively tested to concentrate omega-3 fatty acids from fish oils, under mild enzymatic conditions. However, no lipases with preference for omega-3 fatty acids selectivity have yet been discovered or developed. In this study we performed an exhaustive computational study of substrate-lipase interactions by docking, both covalent and non-covalent, for 38 lipases with a large number of structured triacylglycerols containing omega-3 fatty acids. We identified some lipases that have potential to preferentially hydrolyze omega-3 fatty acids from structured triacylglycerols. However omega-3 fatty acid preferences were found to be modest. Our study provides an explanation for absence of reports of lipases with omega-3 fatty acid hydrolyzing ability and suggests methods for developing these selective lipases.

Co-encapsulation and characterisation of omega-3 fatty acids and probiotic bacteria in whey protein isolate-gum Arabic complex coacervates

Omega-3 fatty acids and probiotic bacteria were co-encapsulated in a single whey protein isolate (WPI)-gum Arabic (GA) complex coacervate microcapsule. Tuna oil (O) and Lactobacillus casei 431 (P) were used as models of omega-3 and probiotic bacteria, respectively. The co-microcapsules (WPI-P-O-GA) and L.casei containing microcapsules (WPI-P-GA) were converted into powder by using spray and freeze drying. The viability of L.casei was significantly higher in WPI-P-O-GA co-microcapsules than in WPI-P-GA. The oxidative stability of tuna oil was significantly higher in spray dried co-capsules than in freeze dried ones.

Exploring omega-3 fatty acids, enzymes and biodiesel producing thraustochytrids from Australian and Indian marine biodiversity

The marine environment harbours a vast diversity of microorganisms, many of which are unique, and have potential to produce commercially useful materials. Therefore, marine biodiversity from Australian and Indian habitat has been explored to produce novel bioactives, and enzymes. Among these, thraustochytrids collected from Indian habitats were shown to be rich in saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs), together constituting 51-76 % of total fatty acids (TFA). Indian and Australian thraustochytrids occupy separate positions in the dendrogram, showing significant differences exist in the fatty acid profiles in these two sets of thraustochytrid strains. In general, Australian strains had a higher docosahexaenoic acid (DHA) content than Indian strains with DHA at 17-31 % of TFA. A range of enzyme activities were observed in the strains, with Australian strains showing overall higher levels of enzyme activity, with the exception of one Indian strain (DBTIOC-1). Comparative analysis of the fatty acid profile of 34 strains revealed that Indian thraustochytrids are more suitable for biodiesel production since these strains have higher fatty acids content for biodiesel (FAB, 76 %) production than Australian thraustochytrids, while the Australian strains are more suitable for omega-3 (40 %) production.