Investigation of Deep-Sea Ecosystems Using Marker Fatty Acids: Sources of Essential Polyunsaturated Fatty Acids in Abyssal Megafauna

Abyssal seafloor ecosystems cover more than 50% of the Earth’s surface. Being formed by mainly heterotrophic organisms, they depend on the flux of particulate organic matter (POM) photosynthetically produced in the surface layer of the ocean. As dead phytoplankton sinks from the euphotic to the abyssal zone, the trophic value of POM and the concentration of essential polyunsaturated fatty acids (PUFA) decrease. This results in pronounced food periodicity and limitations for bottom dwellers. Deep-sea invertebrate seston eaters and surface deposit feeders consume the sinking POM. Other invertebrates utilize different food items that have undergone a trophic upgrade, with PUFA synthesized from saturated and monounsaturated FA. Foraminifera and nematodes can synthesize arachidonic acid (AA), eicosapentaenoic acid (EPA), while some barophylic bacteria produce EPA and/or docosahexaenoic acid. FA analysis of deep-sea invertebrates has shown high levels of PUFA including, in particular, arachidonic acid, bacterial FA, and a vast number of new and uncommon fatty acids such as 21:4(n-7), 22:4(n-8), 23:4(n-9), and 22:5(n-5) characteristic of foraminifera. We suppose that bacteria growing on detritus having a low trophic value provide the first trophic upgrading of organic matter for foraminifera and nematodes. In turn, these metazoans perform the second-stage upgrading for megafauna invertebrates. Deep-sea megafauna, including major members of Echinodermata, Mollusca, and Polychaeta display FA markers characteristic of bacteria, foraminifera, and nematodes and reveal new markers in the food chain.     

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.     

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 (n-3) PUFA and vitamin A Artemia enrichment on pigmentation success of turbot, Scophthalmus maximus (L).

Emulsions with different content of (n-3) PUFA and vitamin A were used to enrich Artemia and examine the effect of these nutrients on pigmentation success in turbot, Scophthalmus maximus (L.). The best pigmentation rates were obtained using an overdose of vitamin A (500 000 IU L^?1), but coincided with a high incidence of skeletal deformations. Higher growth and pigmentation rates were achieved by increasing the quantity of (n-3) PUFA oil in the emulsion. The use of (n-3) PUFA-deficient diet caused the highest occurrence of albinism as well as a cessation of metamorphosis.     

饲料中n-3 PUFA/n-6 PUFA比值对罗氏沼虾幼虾生长性能和抗氧化能力的影响

为探讨饲料多不饱和脂肪酸n-3 PUFA/n-6 PUFA比值对罗氏沼虾幼虾生长性能、虾体肌肉组成、抗氧化能力、血清生理指标以及消化能力的影响,实验设计了n-3 PUFA/n-6 PUFA比值分别为0.37 (D1)、0.59 (D2)、0.93 (D3)、1.51 (D4)和4.38 (D5)的5种等氮等脂饲料饲喂罗氏沼虾幼虾8周,每组设4重复,每个重复40尾虾 。结果显示,饲料n-3 PUFA/n-6 PUFA比值对罗氏沼虾幼虾存活率 (SR)无显著影响;实验虾终末体重 (FW)、增重率 (WGR)和特定生长率 (SGR)随饲料n-3 PUFA/n-6 PUFA比值增加先升后降,均在D3组最高;且D3组虾有最大的肝胰腺蛋白酶活性及脂肪酸合成酶活性。虾体肌肉粗蛋白质、水分和灰分含量不受饲料n-3 PUFA/n-6 PUFA比值影响,但总脂肪含量在D3组显著高于其他组;各组虾体肌肉的n-3 PUFA/n-6 PUFA比值的变化趋势与饲料的n-3 PUFA/n-6 PUFA比值变化趋势呈正相关。随饲料n-3 PUFA/n-6 PUFA比值增加,实验虾血清和肝胰腺中超氧化物歧化酶 (SOD)活性、总抗氧化能力 (T-AOC)和血清铜蓝蛋白 (CP)含量均呈现先升后降趋势,并在n-3 PUFA/n-6 PUFA比值为0.93~1.51时达到最大,但丙二醛 (MDA)含量持续上升;D1组血清总胆固醇 (T-CHO)和甘油三酯 (TG)含量显著高于其他组;血清谷草转氨酶 (AST)和谷丙转氨酶 (ALT)活性先降后升,且D3组最低。研究表明,饲料适宜的n-3 PUFA/n-6 PUFA可显著提升罗氏沼虾生长性能和抗氧化能力,对增重率和特定生长率进行折线回归,建议罗氏沼虾幼虾饲料中最适n-3 PUFA/n-6 PUFA比值为0.86~0.94。     

玉米须萃取物的抗氧化活性

将玉米须乙醇提取物再用氯仿、乙酸乙脂、正丁醇和甲醇分别萃取,获得极性不同的5个组分,比较这5种不同溶剂萃取物在二苯基苦基苯肼(DPPH)体系及在Fe2 诱发卵黄脂蛋白多不饱和脂肪酸(PUFA)过氧化体系中的抗氧化活性。结果表明:在DPPH体系中,正丁醇萃取物的抗氧化活性最强,氯仿萃取物次之;在Fe2 诱发脂蛋白PUFA过氧化体系中,氯仿萃取物的抗氧化活性最强,乙酸乙酯萃取物次之。结果还表明,这5不同溶剂萃取物在不同测试体系中的抗氧化活性顺序不同。     

日粮营养对动物脂肪组织基因表达的影响

脂肪组织是一个强大的分泌器官,它能分泌许多与能量、脂肪代谢相关的酶、激素等.调控能量代谢和脂肪组织生长.本文综述了日粮对动物脂肪组织中基因表达作用的研究状况.并阐述了营养成分对这些基因上下游表达的作用机制.     

PUFA对反刍动物CLA代谢影响的研究进展

共轭亚油酸(Conjugated linoleic acid,CLA)具有诸多生理活性,天然的CLA主要来源于反刍动物产品。在日粮中添加富含多不饱和脂肪酸(Polyunsaturated fatty acid,PUFA)的饲料是提高反刍动物产品中CLA含量的一个重要方法。本文通过PUFA对反刍动物CLA机体、消化道及组织等不同代谢层次的影响进行了综述,提出了目前通过PU-FA提高反刍动物CLA中存在的问题,期望为生产中深入利用反刍动物的高CLA这一优势、提高其产品品质提供帮助。     

Effect of Solvent System on Extractability of Lipidic Components of Scenedesmus obliquus (M2-1) and Gloeothece sp. on Antioxidant Scavenging Capacity Thereof

Microalgae are well known for their biotechnological potential, namely with regard to bioactive lipidic components—especially carotenoids and polyunsaturated fatty acids (PUFA), well-known for therapeutic applications based on their antioxidant capacity. The aim of this work was to evaluate the influence of four distinct food-grade solvents upon extractability of specific lipidic components, and on the antioxidant capacity exhibited against both synthetic (2,2-diphenyl-1-picrylhydrazyl (DPPH^•) and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS^+•)) and biological reactive species (O₂^•− and ^•NO⁻). A eukaryotic microalga (Scenedesmus obliquus (M2-1)) and a prokaryotic one (Gloeothece sp.) were used as case studies. Concerning total antioxidant capacity, the hexane:isopropanol (3:2) and acetone extracts of Sc. obliquus (M2-1) were the most effective against DPPH^• and ABTS^+•, respectively. Gloeothece sp. ethanol extracts were the most interesting scavengers of O₂^•−, probably due the high content of linolenic acid. On the other hand, acetone and hexane:isopropanol (3:2) extracts were the most interesting ones in ^•NO⁻ assay. Acetone extract exhibited the best results for the ABTS assay, likely associated to its content of carotenoids, in both microalgae. Otherwise, ethanol stood out in PUFA extraction. Therefore, profiles of lipidic components extracted are critical for evaluating the antioxidant performance—which appears to hinge, in particular, on the balance between carotenoids and PUFAs.     

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.