Supercritical CO<Subscript>2</Subscript> extraction and concentration of n−3 polyunsaturated fatty acid ethyl esters from tuna cooking juice

Cooking is an important step in the tuna canning process; it usually produces functional solubilized proteins and lipids. The objective of this study was to recover and increase the concentration of functional n−3 fatty acids from the lipids in cooking juice. The lipids contained 12.98% eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) after ethyl esterification and increased to 37.4% with subsequent urea adduct fractionation. Supercritical carbon dioxide extraction was further used to increase the concentration of EPA and DHA ethyl esters. The ratio of EPA+DHA ethyl esters (high-molecular-weight components) to that of C₁₆ and C_18∶1 fatty acid ethyl esters (low-molecular-weight components) was used as a separation index for evaluating the process variables. Experimental results indicated that a high CO₂ density caused a low separation factor. At 1500 psig and 328.2 K, the extraction collected over 600 L CO₂, displaying an accepted concentration factor of EPA+DHA ethyl esters herein. About 80% yield of EPA and DHA was obtained and the ethyl esters increased from 37.4% to 54.3%.     

Effects of food colorants on photooxidation of lipids added to Alaska pollack <Emphasis Type="Italic">Theragra chalcogramma</Emphasis> surimi

Food colorants including rose bengal, erythrosine B, phloxine B and cochineal extracts were added to Alaska pollack Theragra chalcogramma surimi which was previously added with methyl linoleate (MeLe) or ethyl eicosapentaenoate (EPA Et). All the surimi were exposed to a 100-watt light tungsten bulb (4500 lux) in a cold room at 5°C. Four isomeric hydroperoxides, including 13-cis, trans-, 13-trans, trans-, 9-trans,cis- and 9-trans,trans-MeLe hydroperoxides, were generated by oxidation of MeLe added to surimi with and without cochineal extracts after exposure to light for several hours. For the surimi containing rose bengal, erythrosine B and phloxine B, extracted lipids contained not only the above four hydroperoxide isomers but also 12-cis,trans-as well as 10-trans,cis- MeLe hydroperoxides. EPA Et added to surimi which contained rose bengal, erythrosine B and phloxine B generated characteristic isomeric hydroperoxides, including 6-cis,trans- and 17-cis,trans-EPA Et hydroperoxides due to ¹O₂ mediated oxidation. The pro-oxidative potentials of four food colorants in surimi against added EPA Et or MeLe increased as the order of cochineal extracts < phloxine B<erythrosine B<rose bengal. These results clearly demonstrated that the addition of certain food colorants to surimi accelerated ¹O₂ mediated lipid oxidation to result in the generation of characteristic hydroperoxide isomers.     

The effect of docosahexaenoic and eicosapentaenoic acids in live food on the development of abnormal morphology in hatchery-reared brown sole Pseudopleuronectes herzensteini

We examined the effect of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) on the rates of abnormal morphology in juvenile brown sole Pseudopleuronectes herzensteini. Larvae during the D–E stages (15–24 days post hatching) were fed live food containing various amounts of DHA and/or EPA prepared using emulsified oils (DHA ethyl ester, EPA ethyl ester, and corn oil). Larvae during the F–I stages were fed Artemia enriched with a commercial diet supplement. We found that DHA and EPA promoted larval development and improved the incidence of morphological abnormalities in brown sole juveniles to a similar extent. However, DHA was more effective than EPA in preventing the appearance of morphological abnormalities in brown sole. The incidence of normal morphology was clearly improved by an increase of the DHA content in brown sole larvae at 25 days post-hatching. These results suggest that it is important to promote larval development and feed larvae with live food containing high levels of DHA during the D–E stages to prevent morphological abnormalities in brown sole juveniles.     

Dietary Arachidonic Acid Alters Tissue Fatty Acid Profile, Whole Body Eicosanoid Production and Resistance to Hypersaline Challenge in Larvae of the Temperate Marine Fish, Striped Trumpeter (Latris lineata)

We determined the effect of dietary arachidonic acid (20:4n-6, ARA) on tissue ratios of ARA/eicosapentaenoic acid (20:5n-3, EPA) and subsequent whole body production of the eicosanoids, prostaglandin F_2α (PGF_2α) and E₂ (PGE₂) in the marine larvae of striped trumpeter, Latris lineata. Larvae were also subjected to a hypersaline challenge (55 ppt) with an aim to determine possible relationships between tissue fatty acid profiles, prostanoid production, and their tolerance to osmotic challenge. From 5 to 23 days post-hatch (dph) larvae were fed live food, rotifers (Brachionus plicatilis), that had been enriched with one of five experimental emulsions containing increasing concentrations of ARA and constant EPA and 22:6n-3 (docosahexaenoic acid, DHA). Final ARA concentrations in the rotifers were 1.33, 3.57, 6.21, 8.21 and 11.22 mg g⁻¹ DM. Larval growth and survival was unaffected by dietary ARA. Tissue fatty acid concentrations generally corresponded with dietary fatty acids and final tissue ratios of ARA/EPA ranged from 0.9 to 4.9. At 18 and 23 dph whole body concentrations of PGF_2α and PGE₂ generally increased as more dietary ARA was provided in a dose-response manner, and a significant elevation in both PGF_2α and PGE₂ in larvae fed the highest dietary ARA concentration was recorded at 23 dph compared to larvae receiving the lowest concentration. At 18 dph, the highest cumulative inactivity following a hypersaline challenge occurred in larvae fed 8.21 or 11.22 mg ARA g⁻¹ DM, which was significantly greater than those receiving 3.57 mg ARA g⁻¹ DM. At 23 dph no relationship between inactivity of larvae subjected to a hypersaline challenge to dietary ARA was evident. In summary, dietary ARA altered tissue ARA/EPA ratios, prostanoid production and resistance to a hypersaline challenge in larval striped trumpeter. While increasing dietary ARA generally resulted in elevation of prostanoids as well as increasing the number of inactive larvae following a hypersaline challenge at 18 dph, similar trends between prostanoids and larval inactivity were not evident at 23 dph, suggesting the exact mechanisms and relationships between eicosanoids and larval osmoregulation warrants further investigation. Nevertheless the study provides preliminary data on the effect of dietary ARA on the prostaglandin production in marine fish larvae.     

Transgenic modification of the n‐3 HUFA biosynthetic pathway in nibe croaker larvae: improved DPA (docosapentaenoic acid; 22:5n‐3) production

Marine fish are generally unable to produce sufficient quantities of n‐3 highly unsaturated fatty acid (n‐3 HUFA) such as eicosapentaenoic acid (EPA; 20:5n‐3) and docosahexaenoic acid (DHA; 22:6n‐3). Consequently, the seed production of marine fish requires careful nutritional enrichment of live feeds such as rotifers and brine shrimp Artemia to meet n‐3 HUFA requirements for normal growth. Another strategy for improving n‐3 HUFA availability is modifying the biosynthetic pathway of marine fish using transgenic technology. In this study, we conducted a feeding trial with non‐transgenic and transgenic nibe croaker Nibea mitsukurii carrying the elongation of very long‐chain fatty acids protein 2 (Elovl2) gene isolated from masu salmon Oncorhynchus masou and three groups of Artemia (non‐enriched and enriched with two products). For all Artemia groups, docosapentaenoic acid (DPA, 22:5n‐3), which is a direct product of Elovl2, was significantly higher in the transgenic fish than that in non‐transgenic fish, despite the absence of DPA in all diets. Thus, applying transgenic techniques to marine fish at the larval stage are a powerful strategy for modifying n‐3 HUFA biosynthetic pathways.     

Three different lipid sources in formulated start-feeds for turbot (Scophthalmus maximus, L.) larvae – effect on growth and mitochondrial alteration in enterocytes

Turbot, Scophthalmus maximus, larvae were start‐fed with formulated feeds containing soya phospholipids (SP), marine phospholipids (MP) or triacylglycerol (TAG). The levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were the same in the MP and TAG feeds. The control group was given rotifers (R). The larvae were offered feed from day 4 post‐hatch. Larvae fed formulated feed with added MP showed good initial growth and there were no significant differences in weight on day 6 between this group and the group given rotifers. Using feed with added TAG enriched with EPA and DHA gave no growth at all. Using SP as the lipid source in the feed resulted in reduced initial growth. Electron microscopical examination of enterocytes was performed on larval intestines on day 6. Larvae fed MP, TAG or rotifers had normal looking enterocytes with numerous normal looking mitochondria. In the enterocytes of larvae fed SP the mitochondria appeared swollen with a translucent matrix and fragmented cristae. Thus, SP or TAG appear not to be suitable as the sole source of lipids and/or phospholipids in start‐feed for turbot larvae and the effects of MP are not solely caused by high levels of EPA and DHA.     

Changes in oil content,lipid class and fatty acid composition of the microalga Chaetoceros calcitrans over different phases of batch culture

The diatom Chaetoceros calcitrans is a microalgal species used as food for larva in aquaculture for many species worldwide. Chaetoceros calcitrans is an important source of omega 3 long chain (C ≥ 20) polyunsaturated fatty acids (n‐3 LC PUFA), chiefly eicosapentaenoic acid (EPA, 20:5n‐3). This article reports lipid content, lipid class composition and fatty acid profiles of each lipid class during the growth cycle of batch cultures of C. calcitrans. Total lipid content and the concentration of neutral lipid were highest in the late stationary growth phase (day 12). However, the amount of EPA was highest during the logarithmic growth phase (1.24 pg/cell on day 4). EPA was initially concentrated in the glycolipid fraction, but this fraction decreased during logarithmic growth, coinciding with the increase in neutral lipid. Docosahexaenoic acid (22:6n‐3, DHA) (0.91 pg/cell) was reported as a major fatty acid (>10 mg/100 g) in all lipid classes on day 1. DHA was depleted quickly from the neutral lipid and glycolipid classes, but at a slower rate from the polar lipid fraction. This work confirms that C. calcitrans is a good source of lipid, in particular EPA, for larval and adult filter feeders in aquaculture.     

Incorporation and metabolism of 14C-labelled polyunsaturated fatty acids in juvenile gilthead sea bream Sparus aurata L. in vivo

The incorporation, and the capacity for desaturation and elongation in vivo, of intraperitoneally-injected, ¹⁴C-labelled n–3 and n–6 C₁₈ and C₂₀ PUFAs were investigated in juvenile gilthead sea bream, Sparus aurata. The results indicate that juvenile gilthead sea bream have only limited ability to convert CH PUFAs to C₂₀ and C₂₂ HUFAs in vivo. The data are consistent with the results from nutritional studies on larvae, postlarvae and fingerlings that have shown that gilthead sea bream require the provision of preformed eicosapentaenoic and docosahexaenoic acids in the diet. The impairment in the desaturase/elongase pathway was quantitatively and qualitatively similar to that found in turbot, Scophthalmus maximus, being at the level of the 5-desaturase. The low activity of 5-desaturase combined with the consistent finding that arachidonic acid is selectively retained in membrane phosphatidylinositol suggests that, in addition to eicosapentaenoic and docosahexaenoic acids, gilthead sea bream may also have a requirement for preformed arachidonic acid in the diet.Abbreviations AA 5,8,11,14-eicosapenaenoic acid (arachidonic acid, 20:4n–6) - CPL diradyl (diacyl + alkenylacyl + alkylacyl) glycerophosphocholine - DHA 4,7,10,13,16,19-docosahexaenoic acid (22:6n–3) - EPA 5,8,11,14,17-eicosapentaenoic acid (20:5n–3) - EPL diradyl (diacyl, alkenylacyl + alkylacyl) glycerophosphoethanolamine - HUFA highly unsaturated fatty acids ( C₂₀ and with 3 double bonds) - LA 9,12-octadecadienoic acid (linoleic acid, 18:2n–6) - LNA 9,12,15-octadecatrienoic acid (-linolenic acid, 18:3n–3) - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid(s)     

Comparison of Proximate Composition and Fatty Acid Profile of On-Growing and Wild Mediterranean Horse Mackerel (Trachurus mediterraneus)

ABSTRACT

On-growing of horse mackerel is not known in the world. Recently, we have initiated on-growing of the Mediterranean horse mackerel in the Black Sea. Therefore, we aim to compare proximate composition and fatty acid profile of on-growing and wild horse mackerels to evaluate the effect on their nutritional value. Captured horse mackerels less than 13 cm were kept on-growing in sea cages and fed sea bass feed for a year in the southern Turkish Black Sea. Results showed seasonal variations in the proximate contents and fatty acid profile of both on-growing and wild fish groups (p < 0.05). Protein contents of the wild horse mackerel group were significantly higher than the on-growing mackerel group, while the opposite situation was observed for lipid contents (p < 0.05). Despite higher eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels (as fatty acid methyl esters %) of wild horse mackerel in comparison with on-growing group, much higher EPA + DHA contents were accounted for in on-growing fish in the edible portion resulting from higher lipid contents of these samples. The results suggest that lower amounts of fish meat, 50–90 g, from on-growing mackerel would cover the daily suggested value of EPA + DHA; this level is calculated as 51–150 g for wild fish meat.     

Effects of dietary n‐3 PUFA supplements on composition of n‐3 PUFA and expression of fatty acid elongase 5 (AJELOVL5) in sea cucumber,Apostichopus japonicus

This is the first comprehensive study on the effect of dietary polyunsaturated fatty acid (PUFA) levels on the expression of fatty acid elongase 5 (AJELOVL5), PUFA composition, and growth in juvenile sea cucumbers. The specific growth rate (SGR_w) was improved in n‐3 PUFA‐rich diets compared to low n‐3 PUFA diets. AJELOVL5 expression was apparently upregulated in juveniles fed lower PUFA diets relative to higher PUFA diets, with higher expression in the body wall and respiratory tree of juveniles fed diets without ɑ‐linolenic acid (ALA, 18:3n‐3) compared to juveniles fed higher ALA level diets; similar results were also detected in juveniles fed diets with lower eicosapentaenoic acid (EPA, 20:5n‐3), docosahexaenoic acid (DHA, 22:6n‐3), and none of ALA, EPA, or DHA respectively. The concentrations of ALA, EPA, and DHA in tissues were positively related to the content of dietary corresponding PUFA, with higher ALA content in juveniles fed diet ALA_12.71 than in the ALA_7.46 and ALA₀ groups. Similar results were also obtained in sea cucumber fed diets enriched with either EPA or DHA. Interestingly, considerable levels of EPA and DHA were found in the tissues of juveniles fed diets of CK₀ and DHA₀, with no specific input of EPA or DHA, showing that the sea cucumber was capable of biosynthesizing EPA and DHA from their corresponding precursors as ALA and linoleic acid (LA, 18:2n‐6).     

Improved method of protoplast fusion between marine luminescent bacterium and <Emphasis Type="Italic">Escherichia coli</Emphasis> cultivated with fatty acid

An attempt was made to improve the stability toward centrifugation of protoplast fusion between Shewanella sp. and Escherichia coli. Stability of the cell membrane is an important factor in protoplast fusion. In order to change the fatty acid composition of the cell membrane phospholipids, eight fatty acids [caprylic acid, capric acid, palmitic acid, oleic acid, linoleic acid, linolenic acid, eicosapentaenoic acid (EPA) and docosahexaenoic acid] were added to each nutrient medium of Shewanella sp. and E. coli. The protoplasts were treated with lysozyme, and fusion occurred in the presence of a polyethylene glycol solution. The stability of the protoplast of Shewanella sp. decreased after EPA was added to the culture medium, and the stability of the protoplast of E. coli increased after the addition of linoleic acid or linolenic acid. Some fusant colonies that developed on the regenerated medium selected for E. coli with antibiotic tolerance. The efficiency of this fusion was higher than that of initial condition using protoplasts from Shewanella sp. and E. coli incubated without fatty acids. Protoplasts improved the fatty acid composition of the phospholipids. Cell membrane stability can change in order for the weak cells to be taken in by strong cells. These results suggested that the fatty acid composition of cell membrane phospholipids affected the fusant yield of the fusion of these bacteria.     

Effects of dietary eicosapentaenoic acid on growth, survival, pigmentation and fatty acid composition in Senegal sole (Solea senegalensis) larvae during the Artemia feeding period

We examined the effect of dietary eicosapentaenoic acid (EPA, 20:5n‐3) on growth, survival, pigmentation and fatty acid composition of Senegal sole larvae. From 3 to 40 days post‐hatch (dph), larvae were fed live food that had been enriched using one of four experimental emulsions containing graduated concentrations of EPA and constant docosahexaenoic acid (DHA, 22:6n‐3) and arachidonic acid (ARA, 20:4n‐6). Final proportions of EPA in the enriched Artemia nauplii were described as ‘nil’ (EPA‐N, 0.5% total fatty acids, TFA), ‘low’ (EPA‐L, 10.7% TFA), ‘medium’ (EPA‐M, 20.3% TFA) or ‘high’ (EPA‐H, 29.5% TFA). Significant differences among dietary treatments in larval length were observed at 25, 30 and 40 dph, and in dry weight at 30 and 40 dph, although no significant correlation could be found between dietary EPA content and growth. Eye migration at 17 and 25 dph was affected by dietary levels of EPA. Significantly lower survival was observed in fish fed EPA‐H diet. Lower percentage of fish fed EPA‐N (82.7%) and EPA‐L (82.9%) diets were normally pigmented compared with the fish fed EPA‐M (98.1%) and EPA‐H (99.4%) enriched nauplii. Tissue fatty acid concentrations reflected the corresponding dietary composition. ARA and DHA levels in all the tissues examined were inversely related to dietary EPA. This work concluded that Senegal sole larvae have a very low EPA requirement during the live feeding period.     

Fatty acid composition,osmolality, Na+, K+‐ATPase activity,cortisol content and antioxidant status of rainbow trout (Oncorhynchus mykiss) in response to various dietary levels of eicosapentaenoic acid and docosahexaenoic acid

This study was conducted to evaluate the effect of dietary eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels on the fatty acid composition, salinity tolerance and antioxidant status of rainbow trout (Oncorhynchus mykiss). Four diets were formulated with total EPA and DHA contents of 5.41, 9.55, 13.97 and 17.88 g/kg (abbreviated as ED‐5.41, ED‐9.55, ED‐13.97 and ED‐17.88 respectively). Rainbow trout (initial weight of 90.61 ± 9.25 g) were fed the experimental diets for 8 weeks to accumulate significant differences in fatty acid composition and subsequently underwent salinity acclimation. Our results indicated that high dietary EPA and DHA significantly improved the EPA and DHA content in fish tissues. The serum osmolality of fish returned to their freshwater values in the ED‐9.55, ED‐13.97 and ED‐17.88 groups. The Na⁺, K⁺‐ATPase (NKA) activity of fish in the ED‐13.97 group changed dramatically to adapt the fish to the hypertonic environment. Moreover, there was no significant difference in the serum cortisol concentration and liver catalase (CAT) activity of fish in the ED‐13.97 group during salinity acclimation. The liver superoxide dismutase (SOD) activity in the ED‐13.97 group was significantly higher than that in ED‐5.41 and ED‐9.55 groups at the end of salinity acclimation. The muscle malondialdehyde (MDA) content in the ED‐13.97 group was significantly lower than that in the ED‐17.88 group before salinity acclimation and significantly lower than the ED‐5.41 and ED‐17.88 groups on day 7 of acclimation. The results of this study indicate that the rainbow trout in the ED‐13.97 group exhibited optimal salinity acclimation performance.     

A determination of the quantitative requirements for docosahexaenoic acid for juvenile barramundi (Lates calcarifer)

A series of diets with varying docosahexaenoic acid (DHA; 22:6n‐3) inclusion levels (1 g kg^?1 3 g kg^?1, 6 g kg^?1, 10 g kg^?1, 15 g kg^?1 and 18 g kg^?1) were fed to juvenile barramundi (Lates calcarifer) for 6 weeks. Two additional diets examined the addition of arachidonic acid (ARA; 20:4n‐6) or eicosapentaenoic acid (EPA; 20:5n‐3) to the diets at 10 g kg^?1 when DHA was also included at 10 g kg^?1. Fish were fed the diets on a pair‐fed feeding regime to eliminate feed intake variability. Fish were weighed, and blood and tissue samples were collected after 6 weeks. Behavioural parameters were also assessed. Improvement in growth was seen with increasing inclusion of DHA up to a maximum at 10 g kg^?1 inclusion, albeit the response was minor. However, the addition of ARA to the diet reduced the growth response, while the addition of EPA improved the growth response. An improvement in feeding behaviour was also seen with increasing DHA up to a peak at 10 g kg^?1, while those animals fed diets low in DHA showed increasingly cryptic behaviour. With the increasing inclusion of DHA, a range of pathologies were observed, but the addition of an EPA component to the diet limited these pathologies, while the addition of ARA made little improvement and in some cases exacerbated the pathologies.     

不同高温胁迫条件下的坛紫菜中植物激素分析

为了研究高温胁迫条件下内源性植物激素的变化趋势与坛紫菜在高温耐受性之间的相互关系,实验以在宁波象山采集的"浙东1号"坛紫菜叶状体为样品,以液相色—谱质谱联用的选择离子模式作为分析手段,系统地分析了高温胁迫条件下,坛紫菜中11种植物激素含量的变化。结果显示,在高温胁迫环境中共检测出7种植物激素,分别为异戊烯腺苷、异戊烯腺嘌呤、反式玉米素核苷、吲哚乙酸、茉莉酸甲酯、水杨酸和茉莉酸。在20、25、28和35°C下,茉莉酸和茉莉酸甲酯的含量随着刺激温度的上升而升高;吲哚乙酸、反式玉米素核苷、异戊烯腺苷和异戊烯腺嘌呤在高温组中的含量要明显低于对照组,呈现下降的趋势。以28°C作为胁迫温度,在不同胁迫时间下,吲哚乙酸、反式玉米素核苷、异戊烯腺苷和异戊烯腺嘌呤的含量随着胁迫时间的延长而降低;水杨酸和茉莉酸甲酯的含量随着胁迫时间的延长而上升;茉莉酸的含量先升高后下降。在恢复培养过程中,水杨酸、茉莉酸、吲哚乙酸和茉莉酸甲酯的含量均呈现下降、升高又下降的变化趋势,而反式玉米素核苷、异戊烯腺苷和异戊烯腺嘌呤的含量则随着恢复培养过程的进行而逐渐下降。高温胁迫下,坛紫菜在促进生长类激素和胁迫抑制类激素的共同调控下进行胁迫防御,维持自身生长。然而,坛紫菜的防御能力有限,且高温胁迫引起的藻体自损伤可能难以修复。     

壳聚糖对煎烤鱿鱼品质及甲醛生成的影响

为减少煎烤鱿鱼在制作过程中的营养损失和甲醛的生成。实验采用2 g/L的壳聚糖溶液(普通壳聚糖和羧甲基壳聚糖)浸泡处理新鲜巨型枪乌贼(秘鲁鱿鱼)5 min,再进行高温煎烤。甲醛检测的结果显示,煎烤后对照组(纯水浸泡)、壳聚糖组、羧甲基壳聚糖组鱿鱼中的甲醛含量分别为45.56、4.79、8.30 mg/kg,各组之间差异极显著。煎烤前后鱿鱼中几种营养素的检测结果显示,壳聚糖组中天冬氨酸、丝氨酸、脯氨酸、胱氨酸、赖氨酸、十二酸、十五酸、十七酸、十八碳烯酸(n=9)、十八碳烯酸(n=6)、亚油酸(n=6)、二十碳烯酸(n=9)、二十碳二烯酸、二十碳四烯酸、二十碳五烯酸(EPA)、二十二碳五烯酸(n=3)及元素Mg、P、Mn和维生素C含量高于对照组,其他营养素含量较对照组低。羧甲基壳聚糖组中天冬氨酸、丝氨酸、甲硫氨酸、亮氨酸、赖氨酸、组氨酸、十四碳烯酸、十五酸、十六碳烯酸(n=7)、十七酸、花生烯酸(n=9)、二十碳二烯酸、二十碳四烯酸、二十碳五烯酸(EPA)及元素K、Mg、P、Zn、Mn、Se和维生素B含量高于对照组,其他营养素含量较对照组低。另外,EPA及7种人体必需氨基酸在壳聚糖组的含量高于羧甲基壳聚糖组。研究表明,2种壳聚糖均能有效抑制鱿鱼在煎烤过程中甲醛的生成,普通壳聚糖效果更显著,并且能较好地保持鱿鱼的品质。     

Importance of fatty acids in broodstock diets with emphasis on Arctic char (Salvelinus alpinus) eggs

The importance of long-chain polyunsaturated fatty acids, especially the eicosanoid precursors, is addressed in this paper. It has been generally recognized that eicosapentaenoic (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) are of significant importance in fish reproduction while arachidonic acid (AA, 20:4n-6) has often been overlooked. The ratio between C20 fatty acids EPA and AA might be important for many physiological functions depending on the species evolution and its requirements. Arctic char (Salvelinus alpinus) has a much more pronounced freshwater history and therefore different fatty acid requirements than the other commonly farmed salmonids such as salmon (Salmo salar), brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss). Therefore there is reason to formulate a feed that is more suitable for farming of this freshwater species. In this study, freshwater wild-origin char eggs were compared to farmed eggs of char. The ratio n-3/n-6 of total phospholipids of eggs was much lower in the wild fish, 3.5 versus 13.5, and the hatching rate of eggs from natural environment was much higher (20–70% vs. >80%). We conclude that feed based on marine raw product does not fulfill the requirements for essential fatty acids for freshwater char and we suggest that AA is supplemented to the broodstock diet and that at least linoleic acid (18:2n-6) is included in the on-growth diet formulas to lower the n-3/n-6 fatty acid ratio.     

Dietary eicosapentaenoic acid requirement of juvenile rock bream,Oplegnathus fasciatus

A feeding trial was conducted to evaluate the optimum dietary level of eicosapentaenoic acid (EPA, 20:5n‐3) based on growth and non‐specific immune responses in juvenile rock bream. A basal diet without EPA supplementation was used as a control, and six other diets were prepared by supplementing with 4, 8, 12, 16, 20 or 40 g of EPA per kg diet. The actual EPA concentrations of the diets were 0.5, 4.3, 8.5, 13.0, 16.8, 21.0 and 41.2 g of EPA per kg diet, and the diets were abbreviated as EPA_0.5, EPA_4.3, EPA_8.5, EPA_13.0, EPA_16.8, EPA_21.0 and EPA_41.2, respectively. Triplicate groups of fish averaging 1.06 ± 0.01 g (mean ± SD) were fed one of the seven experimental diets at the apparent satiation for 8 weeks. At the end of the feeding trial, weight gain, specific growth rate and feed efficiency of fish fed EPA_16.8, EPA_21.0 and EPA_41.2 diets were significantly higher than those of fish fed EPA_0.5, EPA_4.3, EPA_8.5 and EPA_13.0 diets (p < .05). Superoxide dismutase activity of fish fed EPA_16.8, EPA_21.0 and EPA_41.2 diets were significantly higher than those of fish fed EPA_0.5, EPA_4.3 and EPA_8.5 diets. Fish fed EPA_21.0 and EPA_41.2 diets showed significantly higher lysozyme activity than did fish fed EPA_0.5, EPA_4.3, EPA_8.5 and EPA_13.0 diets. The broken‐line analysis of weight gain indicated that the optimum dietary EPA level was 16.7 g/kg diet. These results suggested that the optimum dietary EPA level in juvenile rock bream could be greater than 16.7 g/kg diet but less than or equal to 16.8 g/kg diet based on the broken‐line analysis and the ANOVA test of weight gain.     

Changes in mitochondrial fatty acid composition following temperature acclimation of carp and their possible effects on FoF1-ATPase activity

Carp undergo temperature acclimation of respiratory function by altering mitochondrial ATP synthase (F_oF₁-ATPase) both quantitatively and qualitatively (Itoi et al. 2003). To address such acclimation temperature-dependent changes of F_oF₁-ATPase activity, we investigated in this study the correlation between the fatty acid composition and F_oF₁-ATPase activity in fast muscle of thermally acclimated carp. The quantities of saturated fatty acids of mitochondria from carp acclimated to 10 °C were significantly lower than those of carp acclimated to 30 °C. While mono- and poly-unsaturated fatty acids tended to increase with cold acclimation of carp, the molar concentration of 16:0 aldehyde in mitochondria from the 10 °C-acclimated carp were less than those from the 30 °C-acclimated fish. The specific activities of F_oF₁-ATPase in the 10 °C- and 30 °C-acclimated fish mitochondria were calculated to be 167±22 and 56±10 nmol/min ⋅ mg mitochondrial protein, respectively, the difference being significant at P<0.005. Taken together, the increase in F_oF₁-ATPase activity in fast muscle mitochondria of carp after cold temperature acclimation may be closely related to the increase of unsaturated fatty acids in mitochondria. Abbreviations: BSA - bovine serum albumin; DHA - docosahexaenoic acid; EGTA - ethyleneglycol bis(2-aminoethylether)tetraacetic acid; EPA - eicosapentaenoic acid; F_oF₁-ATPase - mitochondrial ATP synthase; α-F₁-ATPase - F_oF₁-ATPase α-subunit; β-F₁-ATPase - F_oF₁-ATPase β-subunit; HEPES - 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid; SDS - sodium dodecyl sulfate; SDS-PAGE - SDS-polyacrylamide gel electrophoresis. This revised version was published online in July 2006 with corrections to the Cover Date.     

High n‐3 HUFA levels in the diet of Atlantic salmon affect muscle and mitochondrial membrane lipids and their susceptibility to oxidative stress

Atlantic salmon were fed one of four diets with increasing levels (11–58%of total fatty acids (FAs)) of n‐3 highly unsaturated FAs (HUFAs) in order to investigate the effect on muscle and mitochondrial membrane lipids and their susceptibility to oxidative stress. The high n‐3 eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) groups contained lower percentages of sphingomyelin and cardiolipin in total muscle than the intermediate n‐3 HUFA group. Cardiolipin and sphingomyelin are particularly susceptible to peroxidation, and a reduced percentage of these lipids showed that mitochondrial membranes had been damaged by oxidation. The intermediate n‐3 HUFA group had the highest level of mitochondrial integrity and tendencies of lower thiobarbituric acid reactive substances (TBARS) level than the other dietary groups. The high caspase‐3 activity, in addition to casp3a and bax gene expression levels, in the n‐3 DHA group also suggests that some degree of oxidative stress had occurred. Electron microscopy images showed a higher degree of myofibre–myofibre detachment in fish fed the high HUFA diets than in fish fed the intermediate n‐3 HUFA diet. Our findings show that intermediate levels of n‐3 HUFAs in salmon diets gave the best protection against oxidative damage of mitochondrial membranes and muscle structure.