南极磷虾油中磷脂型脂肪酸的测定方法优化及应用

南极磷虾营养价值高,因富含必需氨基酸、磷脂、虾青素和多不饱和脂肪酸(PUFAs)而具有良好的开发价值.目前,对南极磷虾及其制品脂肪酸含量的分析主要集中在总脂肪酸含量的测定,由于PUFAs不同酯的结合会影响其生物利用度,因此,有必要对南极磷虾不同类型酯的脂肪酸含量进行准确定量.为了明确南极磷虾油中磷脂(PL)型脂肪酸含量...     

Effect of krill phospholipids versus soybean lecithin in microdiets for gilthead seabream (Sparus aurata) larvae on molecular markers of antioxidative metabolism and bone development

The objective of the present study was to compare the effectiveness of dietary marine phospholipids (MPL) obtained from krill and soybean lecithin (SBL) on the rearing performance and development of seabream (Sparus aurata) larvae. Larvae were fed from 16 to 44 day posthatching (dph) five formulated microdiets with three different levels (50, 70 and 90 g kg^–1) of phospholipids (PL) obtained either from an MPL or from a SBL source. Larvae‐fed MPL show a higher survival, stress resistance and growth than those‐fed SBL, regardless the dietary PL level. Overall, the increase in MPL up to 70 g kg^–1 total PL in diet was enough to improve larval gilthead seabream performance, whereas even the highest SBL inclusion level (90 g kg^–1 PL) was not able to provide a similar success in larval growth or survival. Inclusion of SBL markedly increased the peroxidation risk as denoted by the higher TBARs in larvae, as well as a higher expression of CAT, GPX and SOD genes. Moreover, SBL tends to produce larvae with a lower number of mineralized vertebrae and a lower expression of osteocalcin, osteopontin and BMP4 genes. Finally, increasing dietary MPL or SBL lead to a better assimilation of polyunsaturated fatty acids in the larvae, n‐3HUFA (especially 20:5n‐3) or n‐6 fatty acids (especially 18:2n‐6), respectively. In conclusion, MPL had a higher effectiveness in promoting survival, growth and skeletal mineralization of gilthead seabream larvae in comparison with SBL.     

南极磷虾粉脂质及脂肪酸组成分析

为分析南极磷虾粉的脂质成分,实验采用薄层层析法分离制备不同脂质组分,通过酸甲酯化衍生结合气相色谱/质谱法对不同脂质的脂肪酸组成进行比较研究.结果显示:①南极磷虾粉中脂质丰富,总脂含量高达11.37％,且以甘油三酯、磷脂和游离脂肪酸为主,胆固醇含量较低;②总脂脂肪酸主要为C16:0、C18:1n-9、C18:1n-7、C20:5n-3和C22:6n-3,且C20:5n-3和C22:6n-3占总脂肪酸含量的30.67％,表明南极磷虾粉在脂质方面具有较高的营养价值和开发潜力.③不同脂质的脂肪酸组成差异显著,胆固醇酯和甘油三酯中饱和脂肪酸和单不饱和脂肪酸含量均显著高于游离脂肪酸和磷脂(P＜0.05);而游离脂肪酸和磷脂富含多不饱和脂肪酸,两者含量分别为48.50％和49.96％,远高于胆固醇酯和甘油三酯的13.11％和24.36％,具有显著差异(P＜0.05).     

南极磷虾产品研究及发展趋势

南极磷虾生活在南极海域,资源量丰富,具有较高的应用价值.南极磷虾开发耗资巨大,在大规模商业性开发投入之前,做好相关产品与市场分析等基础工作十分必要.本文首先对南极磷虾产品的发展历程进行了分析,继而对南极磷虾产品的主要形式与发展前景进行了剖析,并对主要目标市场进行了定位研究.研究认为：南极磷虾粉和液态磷虾油是目前较为普遍的产品存在形式.其中,磷虾粉可作为蛋白质增强剂、增味剂和重要的氨基酸补充剂使用,可作为渗透调节之用,同时,可以作为矿物质来源、脂质来源、几丁质/几丁聚糖免疫增强剂、天然色素（类胡萝卜素）来源使用.磷虾粉的目标市场更倾向于水产饲料;磷虾油的目标市场主要倾向于营养保健和制药行业.相对于磷虾粉,磷虾油的市场价值更高.     

Biomarkers of bone development and oxidative stress in gilthead sea bream larvae fed microdiets with several levels of polar lipids and α‐tocopherol

Although dietary marine phospholipids are able to improve culture performance of marine fish larvae in a further extend than soybean lecithin, both types of phospholipids (PL) markedly increase oxidative risk. The inclusion of a fat‐soluble antioxidant such as the vitamin E α‐tocopherol could allow a better control of oxidative stress. The objective of this study was to determine the combined effect of graded levels of α‐tocopherol with different levels and sources of krill phospholipids (KPL) and soybean lecithin (SBL) on growth, survival, resistance to stress, oxidative status, bone metabolism‐related genes expression and biochemical composition of sea bream larvae. Sea bream larvae were completely weaned at 16 dph and fed for 30 days seven microdiets with three different levels of PL (0, 40 and 80 g kg^?1 diet) and two of α‐tocopherol 1500 and 3000 mg kg^?1 diet. Sea bream larvae fed diets without PL supplementation showed the lowest survival, growth and stress resistance, whereas increase in PL, particularly KPL, markedly promoted larval survival and growth. However, feeding SBL markedly increased TBARs and GPX gene expression increasing the peroxidation risk in the larvae. Besides, KPL inclusion improved incorporation of n‐3 HUFA and, particularly, EPA into larval tissues, these fatty acids being positively correlated with the expression of BMP‐4, RUNX 2, ALP, OC and OP genes and to bone mineralization for a given larval size class. The increase in dietary α‐tocopherol tends to improve growth in relation to the n‐3 HUFA levels in the diet, denoting the protective role of this vitamin against oxidation. Indeed, dietary α‐tocopherol decreased the oxidative stress in the larvae as denoted by the reduction in larval TBARs contents and gene expression of SOD and CAT, but not GPX. Thus, increase in dietary α‐tocopherol effectively prevented the formation of free radicals from HUFA, particularly EPA, but did not affect the incidence of bone anomalies or the expression of genes related to osteogenetic processes.     

Nutrition of Marine Fish Larvae

ABSTRACT

In the hatchery production of aquatic animals for aquaculture, livefoods such as diatoms; rotifer, Brachionus plicatilis and brine shrimp, Artemia salina, have been used throughout the world. However, such production requires large facilities, maintenance expenses, and labor to produce a desired amount of live foods constantly and reliably. Also, the nutritive value of planktonic organisms is occasionally variable, indicating that the dietary quality of these live foods varies with the content of n-3 highly unsaturated fatty acid (n-3 HUFA). Therefore it is necessary to develop microparticulate diets as a substitute for live foods to further increase theproductivity of seed for fish culture. The nutritional components of microparticulate diets for fish larvae should be determined on the basis of requirements of the larval fish for proteins and amino acids, lipids and fatty acids, carbohydrates, vitamins, and minerals. Moreover, the efficient development of microparticulate diets for the fish larvae has promoted the improvement of nutritional requirement studies. The present reviewconcerning the nutrition of marine fish larvae focuses on the proteins, amino acids, peptides, fatty acids, phospholipids, depigmentation of flatfish, stress tolerance of lipids, incorporation of HUFA in neural tissues, HUFA in egg and larvae, HUFA enrichment of live food, carbohydrates, vitamins, energy source during embryo and larval stages, enzyme supplement in microparticulate diets, and application of microparticulate diets inaquaculture.     

Optimum krill phospholipids content in microdiets for gilthead seabream (Sparus aurata) larvae

The aim of the present study was to determine the optimum dietary levels of krill phospholipids (KPL) for sea bream (Sparus aurata) larvae, and its influence on larval development and digestive enzymes activity. Larvae were fed five formulated microdiets with five different levels of KPL. Complete replacement of live preys with the experimental microdiets for seabream larvae produced high survival and growth rates, particularly in fish fed the highest levels of KPL. In the present study, increase in dietary KPL up to 120 g kg^?1 (100 g kg^?1 total PL) significantly improved larval survival and growth, whereas further increase did not improve those parameters. An increase in alkaline phosphatase, trypsin and lipase activity with the elevation of KPL up to 120 g kg^?1 was also found denoting a better functioning of digestive system. Besides, there was a linear substrate stimulatory effect of dietary KPL on phospholipase A2 activity. Finally, increasing dietary KPL lead to better assimilation of n‐3 HUFA especially eicosapentaenoic acid, reflected in the higher content of these fatty acids in both neutral and polar lipids of the larvae. In summary, KPL were found to be an excellent source of lipids for seabream larvae. Optimum inclusion levels of this ingredient in microdiets to completely substitute live preys at this larval age were found to be 120 g kg^?1 KPL.     

海鱼早期阶段必需脂肪酸和磷脂的研究现状与展望

近年来，海鱼早期阶段必需脂肪酸和磷脂研究取得重要进展。本文以从必需脂肪酸的竞争，DHA、EPA和AA的必需性，饲料脂类的种类与含量，以及海鱼的极性脂-磷脂等方面论述海鱼早期阶段必需脂肪酸和磷脂的研究现状与今后方向。     

A comparison of the fatty acid profiles of adult tissues,and newly hatched,fed and starved Lysmata amboinensis larvae

This study evaluated the lipid content and fatty acid (FA) profile of the hepatopancreas, ovaries and tail muscle of Lysmata amboinensis broodstock, as well as newly hatched larvae subjected to a period of starvation or feed from hatch to Zoea 2. The hepatopancreas had a high lipid content, confirming its role as a process and storage organ in L. amboinensis. Lipids were also a major component of ovarian dry weight, in agreement with reports on other crustaceans during maturation. The tail muscle, being a functional rather than a storage organ, contained low total lipids and was the tissue that closely resembled the FA profile of the newly hatched larvae. Saturated fatty acids (SFAs) and highly unsaturated fatty acids (HUFAs) were the most abundant components of the lipid profiles in broodstock and larvae. The HUFAs docosahexaenoic and eicosapentaenoic were preferentially retained during nutritional stress, confirming their importance for marine cleaner shrimp during early larval development. It appeared polyunsaturated fatty acid and HUFA requirements were met through the larval diet. The SFAs stearic and palmitic were abundant in adult tissues and larvae, whereas monounsaturated fatty acids may have been preferentially catabolized to meet energetic and metabolic larval requirements.     

Antarctic krill (Euphausia superba) meal in the diets improved the reproductive performance of tongue sole (Cynoglossus semilaevis) broodstock

A three‐month feeding trial was conducted to investigate the effects of dietary krill meal on the reproductive performance of tongue sole broodstock. Three diets were formulated to contain different levels of krill meal, 0 (Diet K‐0), 10 (K‐100) and 200 (K‐200) g kg^–1 dry matter, replacing the corresponding contents of protein and lipid from fishmeal and fish oil. Each diet was assigned to triplicate tanks. Compared with K‐0, both K‐100 and K‐200 increased the egg properties such as buoyant eggs rate, egg diameter, oil droplet diameter and SOD activity of the fertilized egg. However, the maturation rate, serum estradiol content, the relative fecundity and hatching rate were improved only by K‐100. The krill meal supplementation increased the concentrations of astaxanthin and n‐3 fatty acids in eggs, but decreased the n‐6 fatty acid concentrations. Both K‐100 and K‐200 increased the survival activity index, but the larval deformity rate was decreased only by K‐100. At 15 DPH, the larval length and the digestive enzyme activities were significantly higher in group K‐200 compared with groups K‐0 and K‐100. In conclusion, dietary supplementation of krill meal, at the level of 100 or 200 g kg^–1 dry matter in exchange of fishmeal and fish oil had the considerable positive effects on C. semilaevis reproduction.     

南极磷虾生物量估算方法

南极磷虾是南大洋生态系统食物网中的关键种,在南大洋中上层生态系统中起着至关重要的连接作用,其资源的养护和持续利用受到国际社会日益关注。磷虾研究已有90多年的历史,其生物量及其分布一直是研究的重点。生物量评估方法也在不断发展,从早期的浮游动物干重法、生产量与生物量比例系数转换法,到网具采样法以及现在流行的声学法,评估手段也变得更为高效。本文通过系统梳理磷虾生物量的主要评估方法,分析其优缺点,并着重从磷虾目标强度模型(TS)参数的海上实测、声学数据中磷虾信号的提取,磷虾单位平方海里声学反射系数(NASC)的计算,基于渔船声学数据修正磷虾渔业单位捕捞努力量渔获量(CPUE),基于渔船声学数据的磷虾集群、洄游和渔场形成机制研究等5个方面对现行生物量声学评估法提出改进建议,提高资源评估的精度,以期为磷虾生物资源的开发与管理提供科学依据。     

Comparison of the lipid profiles from wild caught eggs and unfed larvae of two scombroid fish: northern bluefin tuna (<Emphasis Type="Italic">Thunnus thynnus</Emphasis> L., 1758) and Atlantic bonito (<Emphasis Type="Italic">Sarda sarda</Emphasis> Bloch, 1793)

Lipids and essential fatty acids are determinants of the reproductive process in marine fish, affecting fecundity, egg quality, hatching performance, pigmentation and larval malformation. We have analyzed and characterized the lipids of eggs and unfed larvae of two wild caught scombroid fish, the Atlantic northern bluefin tuna (Thunnus thynnus) and Atlantic bonito (Sarda sarda). Dry matter and total lipid contents, polar and neutral lipid classes and total lipid fatty acid contents were determined in the eggs of bluefin tuna and eggs and unfed larvae during the development of Atlantic bonito. Bluefin tuna eggs had slightly but significantly more dry mass than bonito eggs but very similar lipid content. However, bluefin tuna eggs presented a higher polar lipid content due to increased proportions of phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylinositol (PI). Bonito eggs and larvae showed increasing dry mass and decreasing lipid content with development. The proportion of polar lipids increased due to increased PE, PS and PI, whereas choline-containing polar lipids (phosphatidylcholine and sphingomyelin) remained relatively constant. Free cholesterol also increased, whereas the levels of other neutral lipids, especially triacylglycerol and steryl ester fractions, decreased, presumably due to utilization for energy to drive development. Bluefin tuna eggs had higher levels of n − 3 and n − 6 highly unsaturated fatty acids due to higher docosahexaenoic and arachidonic acid contents, respectively, than bonito eggs. The results are discussed in relation to the lipid and fatty acid requirements of larval scombroid fish in comparison to those of other larval marine finfish species under culture conditions.     

Preparation,Gel Electrophoresis Analysis,and Nutritional Evaluation of a Functional Krill Protein Concentrate with Low Fluoride Level from Antarctic krill (Euphausia superba)

A functional krill protein concentrate (FKPC) with low level of fluoride was prepared from frozen Antarctic krill (Euphausia superba) using isoelectric precipitation and phosphoric acid treatment. Variations in proteins and nutritional properties of the FKPC were analyzed. Gel electrophoresis analysis indicated that some variations in protein composition were introduced by autolysis and pH shift during the process of isoelectric precipitation and fluoride removal. The FKPC was composed of 81.01% ± 0.45% crude protein (dry basis) and 16.78% ± 0.26% total lipid (dry basis), exhibited excellent amino-acid composition in all the essential amino acids, exceeding the requirements of the FAO/WHO/UNU for food protein, and was rich in omega-3 polyunsaturated fatty acids, especially eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids present in phospholipids. The fluoride level of FKPC was 79.79 ± 2.10 μg/g (dry basis), which met the USFDA requirements. The surimi yield reached 43.74%, and the mean recovery rate of protein and fat was 52.68% and 41.44%, respectively. The nutritional value, safe level of fluoride, and high yield rate provide evidence that FKPC has a promising future in the food industry as a new aquatic protein and lipid resource.     

Fatty acids as trophic biomarkers in vitellogenic females in an impounded tropical river

Fatty acids have been used in marine biogeochemistry as food chain biomarkers, but in freshwater these studies are rare. In order to evaluate the fatty acid potential as biomarkers in freshwater, their profile was analyzed during vitellogenesis in two fish species, in both waterfall and reservoir environments of the Paraíba do Sul River Basin. Detrivorous Hypostomus affinis and omnivorous Geophagus brasiliensis seem to elongate and desaturate polyunsaturated fatty acids (PUFA) and transfer them to the ovaries’ phospholipids. Waterfall Geophagus brasiliensis have more highly unsaturated fatty acids in the liver, but in the reservoir, accumulation mainly occurs in muscle and ovary triglycerides, suggesting trophic opportunism and a plasticity during vitellogenesis. In Hypostomus affinis, PUFA alteration occurs only in the reservoir, suggesting a high phytoplankton occurrence. Eutrophication and water speed is reflected in Hypostomus affinis ovaries by higher PUFAn3 and bacterial fatty acids. As in marine environments, analysis of mono- and polyunsaturated fatty acids during vitellogenesis can be used as a tool in food chain studies in freshwater.     

Fatty acids of the lipids of cultured herring

The composition of fatty acids was determined in neutral lipids and phospholipids from cultured herring reared on a diet based on squid muscle. The phospholipids retained the fatty acid pattern characteristic of marine fish. In contrast, the fatty acids of neutral lipids tended to reflect the fatty acid spectrum in the diet, and were very different from the characteristic pattern of herring oils. It was concluded that the cultured herring had little ability to modify dietary fatty acids and that the enzymes of phospholipid metabolism possess considerable specificity for certain fatty acids.     

Nutrient Compositions and Properties of Antarctic Krill (Euphausia superba) Muscle and Processing By-Products

ABSTRACT

This study investigated the morphometrics, proximate chemical compositions, pH, total amino acid (TAA), fatty acid profile, and minerals of the processing by-products of Antarctic krill (Euphausia superba). The nutrient compositions and properties of the by-products were revealed by being compared to those of Antarctic krill muscle and the economically important species of freshwater prawns (Macrobrachium nipponense) and penaeid shrimps (Metapenaeus ensis). The by-products are worthy of utilization because of the high ratio to the total weight (65.7%). The crude protein contents in the muscle and by-products of krill are 17.4 and 11.7%, respectively. The krill proteins have higher contents of essential amino acids (EAAs). The EAAs constitute 42 and 37% of the TAAs in muscle and by-products, respectively. The krill processing by-products contain high levels of total lipid (3.3%), and polyunsaturated fatty acids constitute 34% of fatty acids with high levels of eicosapentaenoic acid (EPA; 19.08%) and docosahexaenoic acid (DHA; 10.02%). Krill meat provides considerable iron, zinc, calcium, selenium, and copper. It is imperative to lower the fluoride level (70.1 mg/kg, wet basis) in krill muscle. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) reveals that myosin heavy chain (MHC) and actin are the major proteins in muscle, and their contents vary between species.     

Biochemical and fatty acid dynamics during larval development in the razor clam Ensis arcuatus (Bivalvia: Pharidae)

This article reports on the changes in gross biochemical and fatty acid composition of the razor clam Ensis arcuatus larvae throughout development. Protein was the largest component of dried larval tissues. The energy required for embryogenesis in E. arcuatus oocytes was obtained from stored proteins and carbohydrates, while total lipids increased significantly. Lipids and carbohydrates have an important role as energy contributors from day 1 to day 8. During larval development the strategy was to indistinctly store energy reserves (protein, lipid and carbohydrate) to surpass metamorphosis. During embryonic development there was a gain in fatty acids of neutral and polar lipids. A depletion of fatty acids in neutral lipids was observed from day 1 to day 8 in E. arcuatus larvae, mainly due to the decrease in 16:0 and EPA. NMID fatty acids were present in amounts similar to those of some PUFAs in polar lipids, especially 22:2NMID.     

Influence of dietary polar lipids' quantity and quality on ingestion and assimilation of labelled fatty acids by larval gilthead seabream

Dietary supplementation of phospholipids seems to be extremely important to promote growth and survival in fish larvae. Several studies also suggest the importance of n-3 highly unsaturated fatty acids (HUFA) rich phospholipids to further enhance larval performance. In the present study, four different diets were formulated in order to compare the effect of total dietary polar lipid contents, of soya bean lecithin supplementation and of feeding n-3 HUFA in the form of neutral or polar lipids on ingestion and incorporation of labelled fatty acids in gilthead seabream larvae. These diets were prepared including radiolabelled fatty acids from palmitoyl phosphatidylcholine, glycerol trioleate, free oleic acid (FOA) and free eicosapentaenoic acid (FEPA) and were fed to 25 day-old larvae. The results of these experiments showed that the elevation of the dietary polar lipid levels significantly improved microdiet ingestion, regardless of the origins of the polar lipids. This effect caused an improved incorporation of phosphatidylcholine fatty acids to the larval polar and total lipids (TL) as the dietary polar lipids increased. Nevertheless, a better incorporation of fatty acids from dietary polar lipids in comparison with that of fatty acids from dietary triglycerides into larval lipids was found in gilthead seabream, whereas a better utilization of dietary triglycerides fatty acids than dietary free fatty acids could also be observed. Besides, the presence of n-3 HUFA rich neutral lipids (NL) significanlty increased the absorption efficiency of labelled oleic acid from dietary triglycerides, but the presence of n-3 HUFA rich polar lipids, particularly improved the incorporation of FEPA. This fatty acid was preferentially incorporated into larval polar lipids in comparison with FOA.     

南极磷虾脂溶性成分的研究进展

南极磷虾脂类营养含量丰富:油脂提取物中二十碳五烯酸(EPA)含量高于15%,二十二碳六烯酸(DHA)含量高于9%,总磷脂含量高于20%,冰鲜的南极磷虾中总虾青素含量高于3 mg/100 g。本文总结了近年来国内外学者对南极磷虾油中的脂肪酸、虾青素、脂溶性维生素、甾醇等脂溶性成分及其功效的研究进展,以及磷虾油提取方法、多种成分稳定性的研究结果,并展望了我国开展南极磷虾脂溶性成分综合利用的前景。     

南极磷虾粉营养成分的分析与比较

对两种不同来源的南极磷虾粉营养成分进行了分析和评价,并与进口鱼粉的营养成分进行了比较。结果显示,上海开创远洋渔业有限公司开利轮产南极磷虾粉(磷虾粉S)粗蛋白含量为62.89%,高于辽宁远洋渔业有限公司安兴海轮产南极磷虾粉(磷虾粉D)和进口鱼粉的粗蛋白含量,两种磷虾粉粗蛋白质含量分别达到一级品和二级品指标。磷虾粉S、磷虾粉D和进口鱼粉18种氨基酸总量分别为66.36%、57.19%、67.24%,鲜味氨基酸含量较高分别为24.65%、20.69%和25.25%,8种必需氨基酸含量分别为31.43%、21.74%、25.62%,且磷虾粉S必需氨基酸含量明显高于磷虾粉D和鱼粉,磷虾粉S、磷虾粉D和进口鱼粉三种样品必需氨基酸指数(EAAI)分别为57.83、49.89、57.07;磷虾粉S、磷虾粉D和进口鱼粉三种样品脂肪酸中,多不饱和脂肪酸含量较高分别为43.28%、40.25%、46.58%,且磷虾粉S的EPA与DHA的总量(36.14%)高于磷虾粉D(31.30%)及鱼粉(27.65%)。分析结果表明,南极磷虾粉蛋白含量高,且氨基酸组成符合FAO/WHO推荐的蛋白质理想模式,含有丰富的多不饱和脂肪酸,营养价值较高,具有较大的开发利用前景。