A 13CO2 Enrichment Experiment to Study the Synthesis Pathways of Polyunsaturated Fatty Acids of the Haptophyte Tisochrysis lutea

The production of polyunsaturated fatty acids (PUFA) in Tisochrysis lutea was studied using the gradual incorporation of a ¹³C-enriched isotopic marker, ¹³CO₂, for 24 h during the exponential growth of the algae. The ¹³C enrichment of eleven fatty acids was followed to understand the synthetic pathways the most likely to form the essential polyunsaturated fatty acids 20:5n-3 (EPA) and 22:6n-3 (DHA) in T. lutea. The fatty acids 16:0, 18:1n-9 + 18:3n-3, 18:2n-6, and 22:5n-6 were the most enriched in ¹³C. On the contrary, 18:4n-3 and 18:5n-3 were the least enriched in ¹³C after long chain polyunsaturated fatty acids such as 20:5n-3 or 22:5n-3. The algae appeared to use different routes in parallel to form its polyunsaturated fatty acids. The use of the PKS pathway was hypothesized for polyunsaturated fatty acids with n-6 configuration (such as 22:5n-6) but might also exist for n-3 PUFA (especially 20:5n-3). With regard to the conventional n-3 PUFA pathway, Δ6 desaturation of 18:3n-3 appeared to be the most limiting step for T. lutea, “stopping” at the synthesis of 18:4n-3 and 18:5n-3. These two fatty acids were hypothesized to not undergo any further reaction of elongation and desaturation after being formed and were therefore considered “end-products”. To circumvent this limiting synthetic route, Tisochrysis lutea seemed to have developed an alternative route via Δ8 desaturation to produce longer chain fatty acids such as 20:5n-3 and 22:5n-3. 22:6n-3 presented a lower enrichment and appeared to be produced by a combination of different pathways: the conventional n-3 PUFA pathway by desaturation of 22:5n-3, the alternative route of ω-3 desaturase using 22:5n-6 as precursor, and possibly the PKS pathway. In this study, PKS synthesis looked particularly effective for producing long chain polyunsaturated fatty acids. The rate of enrichment of these compounds hypothetically synthesized by PKS is remarkably fast, making undetectable the ¹³C incorporation into their precursors. Finally, we identified a protein cluster gathering PKS sequences of proteins that are hypothesized allowing n-3 PUFA synthesis.     

国内外水生态分区研究进展

水生态分区研究自开展以来已有半个世纪的历史,取得了大量的研究成果,发展出为不同管理目标服务的多种类型、指标体系和研究方法。总结了国内外关于水生态分区研究的概念框架以及中国、美国和欧洲水生态分区的研究概况,综合阐述了典型水生态分区的指标体系和常见研究方法如生物评价法和统计分析法等,最后概括分析了当前国内外水生态分区在水环境管理应用中存在的普遍问题和困难。     

Effect of three diets on the gametogenic development and fatty acid profile of Paracentrotus lividus (Lamarck, 1816) gonads

In this study, the effects of three diets were investigated to enhance Paracentrotus lividus production for commercial purposes. P. lividus were fed ad libitum for 80 days with: diet A—fresh Codium tomentosum Stackhouse, 1797; diet B—formulated using a jellified mix of macroalgae and vegetables, including C. tomentosum (20%), Coralina sp. Linnaeus, 1758 (17%), cabbage Brassica oleracea var. capitata Linnaeus, 1753 (30%), carrot Daucus carota Linnaeus, 1753 (30%) and agar (3%) as a gelling agent. Diet C consisted of maize Zea mays Linnaeus, 1753 (56%) and New Zealand spinach Tetragonia tetragonoides (Pallas, 1781) Kuntze, 1891 (44%). Their effects on the gonadal and somatic growths, gonadosomatic index (GI) and gametogenesis were evaluated, as well as on the total lipid content and fatty acid composition of sea urchin's gonads. Diet A provided high values of eicosapentaenoic acid (EPA). Gonads of sea urchins fed with diet A were found mostly in growth and maturation stages of gametogenesis and showed the lowest lipid content. Sea urchins fed with diet B presented their gonads in the reabsorption stage and had the highest values of omega‐3 polyunsaturated fatty acids (PUFAs). Sea urchins fed with diet C were in the early stages of gametogenesis and had the highest values of lipid content, plus omega‐6 PUFAs. Once as an ingredient in a balanced mix with vegetables, C. tomentosum can be a key factor to the development of new promising high‐quality and low‐cost feed for P. lividus roe enhancement.     

Enhancement of gilthead seabream (Sparus aurata) larval growth by dietary vitamin E in relation to two different levels of essential fatty acids

The objective of this study was to determine the effect of dietary vitamin E on gilthead seabream (Sparus aurata) growth and survival, at two different highly unsaturated fatty acids (HUFAs) levels. Eighteen days old gilthead seabream larvae were fed four formulated experimental diets combining two different dietary levels of HUFAs (M: medium 2.5 + 1.5, DHA + EPA, H: high 5 + 2.5 DHA + EPA g per 100 g) with two different levels of vitamin E (M: medium 540 mg kg^?1, H: high 2900 mg kg^?1): MM, MH, HM, HH (HUFA/vitamin E). After 2‐week feeding trial, the average survival rate was 52.6% and there were no significant differences found among treatments. Increase in vitamin E up to high level markedly improved larval growth, particularly when dietary HUFA levels were lower, suggesting a higher protection value when these fatty acids are more limiting. At medium dietary HUFA levels, increase in vitamin E from medium to high level enhanced larval growth performance in terms of total length. Moreover, increase in vit E enhanced HUFAs content in the larval polar lipids denoting the anti‐oxidative effect of vitamin E.     

Enhancing highly unsaturated ω‐3 fatty acids in phase‐fed rainbow trout (Oncorhynchus mykiss) using Alaskan fish oils

Rainbow trout, average weight 185–187 g, were fed feeds containing menhaden oil, canola oil or fish oils (pollock, pink salmon or rockfish) produced from Alaskan seafood processing waste as the added oil for 8 weeks, at which time the fish weighed 391–411 g (average 404 g, pooled SE = 5.7). The fish were previously fed from 75 g average weight fed commercial feed containing poultry oil as the added oil. No significant differences were measured in final weight or feed conversion ratio among dietary treatment groups. Significant differences were found in fillet ω‐3 fatty acid (FA) levels from fish receiving fish oil‐supplemented feeds compared to those from fish receiving feeds containing canola oil. Fillet contents of eicosapentaenoic acid (EPA; 20:5ω3) and decosahexaenoic acid (DHA; 22:6ω3) were highest in the pollock oil treatment group, although all fish oils increased highly unsaturated ω‐3 FA contents (mg 100 g^?1) of fillets. Fish oil used through the production cycle was reduced by 25% by supplementing feeds with poultry oil during the middle phase of production (75–175 g) compared to using feeds containing fish oil throughout the production cycle. Fish oils recovered from Alaskan seafood processing waste were suitable alternatives to conventional fish oil as ingredients in rainbow trout production feeds.     

杂交鲶(怀头鲶♀×鲶鱼♂)及其亲本肌肉营养成份分析

对怀头鲶♀×鲶鱼♂杂交F1代和亲本的肌肉中蛋白质、脂肪、水份、氨基酸及其脂肪酸中EPA、DHA含量进行了较全面的分析。结果表明：虽杂交鲶粗蛋白含量（17.28％）略高于亲本怀头鲶（16．78％）、鲶鱼（16．82％）,但粗脂肪含量高于亲本平均含量的4．3倍。EPA、DHA含量也明显超过亲本。3种鱼肌肉中含有18种氨基酸,杂交鲶肌肉中的THR、VAL、MET、IYS、TRY及有鲜味的ASP、GLU、ALA的含量也较突出。     

温度影响三角褐指藻生长和脂肪酸组成的初步探讨

实验对指数生长期、指数生长末期、静止期和衰亡期的三角褐指藻脂肪酸进行研究,发现在指数生长末期二十碳五烯酸(Eicosapentaenoic acid,EPA)含量最高。实时定量PCR分析三角褐指藻Δ5脂肪酸去饱和酶基因在不同生长时期的表达变化水平差异显著,其中指数生长期表达量最高。为了探讨温度对微藻生长和脂肪酸组成的影响,作者在不同温度下对三角褐指藻进行了培养,并测定其生长和脂肪酸组成。结果表明,三角褐指藻的生长随着温度的升高到达指数生长末期所需时间缩短;不同温度下三角褐指藻的主要脂肪酸为C14∶0、C16∶0、C16∶lΔ9、Cl8∶lΔ9、C18∶2Δ9,12、C20∶5n3(EPA),总脂肪酸随着温度的升高依次呈现先升高后降低的趋势,在15℃时EPA占总脂肪酸的百分含量达到最高值25.71%。     

Use of marine yeasts as an available diet for mass cultures of Moina macrocopa

A 4‐week feeding trial was conducted to test the use of marine yeasts purified from seawater and sediments as a dietary source in cultivating a cladoceran, Moina macrocopa, a potential alternative live food for fish larvae. Optimal growth conditions of two yeast strains were obtained for NaCl concentration, pH and temperature. Moina macrocopa was cultivated using different diets: marine yeasts (Debaryomyces hansenii Yeast‐14 and Candida austromarina Yeast‐16) and a commercial diet (Erythrobacter sp. Sπ‐I). The essential amino acids of both the yeast strains were nearly as great as those in M. microcopa. Further, the yeast‐fed M. macrocopa had essential amino acid profiles similar to the documented values for rotifers and Artemia enriched in microalgae and commercial diets. Erythrobacter sp. Sπ‐I lacked n‐3 polyunsaturated fatty acids, 20:5n‐3 and 22:6n‐3, which were also low but detected in both yeasts. An increase in the 20:5n‐3 [eicosapentaenoic acid (EPA)] and 22:6n‐3 [docosahexaenoic acid (DHA)] levels, compared with the levels in yeast strains themselves, was more pronounced in the 22:6n‐3 level of Moina fed the C. austromarina, resulting in a high DHA:EPA ratio. When the Moina diets were switched, their δ¹³C values shifted gradually towards the values of the switched diets. Diet switch from Erythrobacter sp. Sπ‐I to C. austromarina Yeast‐16 resulted in a more rapid turnover of Moina tissue carbon than that in the inverse case. When fed a mixed diet, the δ¹³C values of Moina tissue approached the value of marine yeasts immediately. These temporal changes in the δ¹³C values of Moina tissue indicate the preferential ingestion of marine yeasts and a selective assimilation of the carbon originated from marine yeasts. These findings suggest that marine yeasts, particularly C. austromarina Yeast‐16, are highly available to mass cultures of M. macrocopa, providing better nutritional and dietary values than the commercial diet (Erythrobacter sp. Sπ‐I).     

Fatty acid composition of New Zealand green-lipped mussels, Perna canaliculus: Implications for harvesting for n-3 extracts

Marine bivalves offer a potentially important source of long-chained polyunsaturated fatty acids (PUFA) for human health supplements. Lipid extracts from individual New Zealand green-lipped mussels (NZGLM) were analyzed as fatty acid methyl esters (FAME) by gas chromatography to assess geographical and seasonal differences between large (86 ± 1 mm) male and female and small (44 ± 1 mm) mussels. PUFAs dominated in spring and summer, comprising ∼ 50% of total fatty acids. Moreover, the commercially important n-3 fatty acids, 20:5n-3 (eicosapentaenoic acid, EPA) and 22:6n-3 (docosahexaenoic acid, DHA) together accounted for 70-79% of total PUFAs in spring and summer. During winter there was a marked decrease in condition and total n-3 PUFAs and a concomitant increase in saturated fatty acids in mussels, suggesting they had already spawned, had increased metabolic demands and limited PUFA-rich phytoplankton as food. While total n-3 content was not significantly different, there were geographical differences in individual n-3 fatty acids. Mussels collected from the cooler waters of Stewart Island had greater levels of 20:5n-3 (EPA), while those collected in Marlborough had greater concentrations of 22:6n-3 (DHA), which was attributed mainly to differences in phytoplankton composition. Total n-3 content and the condition index varied seasonally with greater concentrations of n-3 PUFAs, especially EPA, recorded in large mussels in spring, coincident with spring diatom blooms. Total PUFA levels and condition indices remained high in summer. There was no significant difference in condition indices, total n-3 content, DHA or EPA levels between large male and female mussels. Conversely, large mussels had significantly greater amounts of n-3 PUFAs than small mussels at Marlborough Sounds, while small mussels had marginally greater total n-3 concentrations than large mussels at Stewart Island. Taken together, these results suggest that the NZGLM offers a potentially important source of n-3 PUFAs for human health supplements. Our findings suggest that optimal harvesting conditions occur in spring when mussel condition and n-3 content peak for large mussels. Although DHA and EPA levels varied geographically, total n-3 content was not significantly different between sites, which implies that harvesting mussels for n-3 extracts would be driven more by logistical considerations.