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.     

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。     

饲料中脂肪水平对海湾扇贝性腺发育、脂肪酸组成和组织结构的影响

为了探讨配合饲料及其脂肪含量对海湾扇贝性腺发育、脂肪酸组成和组织结构的影响,在基础饲料中通过梯度添加鱼油（质量分数为0%、3%、6%和9%）配置4种脂肪质量分数为7%、10%、14%和17%的等氮饲料（质量分数为47%粗蛋白）,分别记为F7、F10、F14和F17,并以小新月菱形藻（Nitzschia closterium f. minutissima）作为对照组,试验在室内1 000 L的水槽中进行,每种饲料随机投喂3组扇贝[初始体质量为（40.79±1.70） g],每个重复30只,试验周期30 d。结果显示：饲料中脂肪含量未显著影响扇贝死亡率（P>0.05）,对照组死亡率显著低于各个饲料处理组（P<0.05）。性腺指数（gonadosomatic index,GSI）随着饲料脂肪含量增加呈先上升后下降的趋势,在F14组GSI达到最高,与对照组差异不显著（P>0.05）,显著高于初始时期（P<0.05）。性腺中雌性部分含水量F10组最高,显著高于F17组（P<0.05）;雄性部分各处理组间差异不显著（P>0.05）。性腺中雌性部分粗脂肪含量显著高于雄性部分（P<0.05）。性腺中脂肪酸的含量PUFA>MUFA>SFA,饲料中脂肪含量显著影响性腺中SFA、MUFA、n-3PUFA、DHA和EPA的含量（P<0.05）,当饲料中脂肪含量为14%或17%时达到最高值。性腺组织切片可见,各个处理组卵母细胞和精原细胞充满整个滤泡腔,滤泡密集,滤泡之间无空隙。由此可得出,配合饲料替代部分单胞藻饲喂海湾扇贝,可使其性腺发育至育苗要求,饲料中脂肪质量分数为14%时可以代替部分单胞藻饲喂海湾扇贝促进性腺发育。     

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.     

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.     

饲料中添加鱼油对肥育猪脂肪酸组成的影响

在肥育猪饲料中添加鱼油生产富含DHA(docosahexaenoic acid),EPA(eicosapentaenoic acid)的猪肉.结果表明:在肥育猪饲料中添加2.5鱼油,可以极显著提高脂肪、肌肉中的DHA,EPA含量.100 g肌肉中EPA对照组为4.7 mg,鱼油组为30.4 mg,是对照组的6.5倍;DHA对照组为13.5 mg,鱼油组为53.8 mg,是对照组的4倍,证明在饲料中添加鱼油生产富含DHA,EPA的猪肉可行.在肥育猪饲料中添加鱼油对猪生产性能、肉质均没有影响.     

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.     

Nano- and Microdelivery Systems for Marine Bioactive Lipids

There is an increasing body of evidence of the positive impact of several marine lipids on human health. These compounds, which include ω-3 polyunsaturated fatty acids, have been shown to improve blood lipid profiles and exert anti-inflammatory and cardioprotective effects. The high instability of these compounds to oxidative deterioration and their hydrophobicity have a drastic impact in their pharmacokinetics. Thus, the bioavailability of these compounds may be affected, resulting in their inability to reach the target sites at effective concentrations. In this regard; micro/nanoparticles can offer a wide range of solutions that can prevent the degradation of targeted molecules, increase their absorption, uptake and bioavailability. In this work we will present the options currently available concerning micro- and nanodelivery systems for marine lipids; with emphasis on micro/nanoparticles; such as micro/nanocapsules and emulsions. A wide range of bottom-up approaches using casein, chitosan, cyclodextrins, among others; will be discussed.     

基于农药毒力计算的6个统计分析软件比较

选用了能够用于农药毒力数据机率值分析的通用软件(SAS、SPSS和DPS)和专门程序(Polo、BA和EPA机率值分析程序),以二化螟对三唑磷的室内毒力数据为例,比较分析不同统计分析软件的计算结果。结果表明,当对照组死亡率为0时,6个软件程序计算得到的LD值(LD5、LD50和LD95)、截距和斜率等基本相等,但是LD值的95%置信限有差异。当对照组死亡率不为0时,发现6个程序计算得到的毒力资料数值都有差异,其中LD(LD5、LD50和LD95)、截距和斜率的数值相差较小,但是LD值的95%置信限差异较大。     

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.