Effects of reactive radicals and heat on <Emphasis Type="Italic">trans</Emphasis>-isomerization of eicosapentaenoic acid

Trans geometric isomers of eicosapentaenoic acid (TEPA) have been found as minor components in human platelets. However, there is little information on the mechanism of trans-isomerization of eicosapentaenoic acid (EPA) in vitro and in vivo. The effects of reactive radicals and heat on trans-isomerization of EPA were examined. Trans-isomerization occurred when EPA ethyl ester reacted with nitrogen dioxide radical (NO₂) but not with 2,2′-azobis(2,4-dimethylvaleronitrile). TEPA was also produced from EPA ethyl ester heated at 200°C for 60 h. No TEPA, however, was detected in sardines Sardinops melanostictus after boiling, roasting, or microwave heating. These results suggest that EPA is trans-isomerized by NO₂ in vivo while trans-isomerization of EPA does not occur during conventional cooking.     

Effect of Squid Melanin-Free Ink and Pre-Emulsification on Properties and Stability of Surimi Gel Fortified with Seabass Oil during Refrigerated Storage

ABSTRACT

The effects of melanin-free ink (MFI) and pre-emulsification on gel properties and stability of bigeye snapper surimi gel fortified with seabass oil during refrigerated storage of 10 days were studied. Lipid oxidation as determined by peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) of surimi gel increased as the level of seabass oil increased (P < 0.05). When MFI was incorporated into surimi gel, lower PV was obtained throughout the storage (P < 0.05). Addition of seabass oil pre-emulsified with soy protein isolate (SPI) in the presence of MFI yielded surimi gel with the highest breaking force and could improve oxidative stability during refrigerated storage (P < 0.05). Slight decrease in whiteness was found in surimi gel added with MFI, while those added with MFI along with pre-emulsified seabass oil showed increased whiteness (P < 0.05). Addition of MFI did not affect total viable count and psychrophilic bacterial count in surimi gels. Thus, the incorporation of pre-emulsified seabass oil prepared using SPI in conjunction with MFI could improve quality and oxidative stability of gel from bigeye snapper surimi.     

Effect of Bay (Laurus nobilis L.) Essential Oil on Surimi Gels Nutritionally Enhanced with Salmon and Flaxseed Oils

Bay essential oil (BEO) at 0.5 and 1 g/100 g was added to surimi gels fortified with omega (ω)-3 rich oils. Although the fortification resulted in increased (P < 0.05) susceptibility of surimi to lipid oxidation, 0.5 g/100 g BEO significantly reduced TBARS (P < 0.05). Addition of ω-3 rich oil or BEO improved the texture. Color values of the surimi seafood were improved particularly when BEO was added. Sensory properties, water activity, and pH values were generally within the acceptable range. These results suggest that the incorporation of BEO may allow food manufacturers to nutritionally enhance surimi seafood with beneficial ω-3 rich oils.     

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.     

Rheology and Texture Properties of Surimi Gels of Northern Snakehead (Channa Argus) as Affected by Angelica Sinensis (Oliv.) Diels. (Danggui) Powder

Northern snakehead (Channa argus) is very popular freshwater fish in China and is expected to become a new surimi material with therapeutic efficacy. The Angelica sinensis powder is a type of famous Chinese medicine that helps to regulate metabolism and enhance the immune function. Therefore, an upmarket medicated surimi product with hygienic qualities was designed to adapt to the stressful current lifestyle using snakehead and Angelica sinensis powder. The texture, whiteness, rheological properties, and sensory properties were studied to further optimize the dosage of the Angelica sinensis powder used in the surimi processing. The results indicated that the maximum breaking force and breaking distance were obtained at a concentration of 4 g/kg, and the whiteness value and driage were significantly decreased with the increase of the amount of the Angelica sinensis powder. Ultimately, the highest total sensory evaluation score was obtained for samples with the addition of 2 g/kg Angelica sinensis powder. This study shows that the Angelica sinensis powder has the potentiality to become an added ingredient to snakehead surimi products in order to improve the health status of sub-health population.     

Nutritional Composition and Thermal Properties of Goldstripe Sardinella (Sardinella gibbosa) Fillets and By-Products

Fish is a nutritious food for the whole family and a good source of protein and n-3 fatty acids. The fish-processing industry produces tonnes of fish-processing waste and by-products annually. Disposal of untreated by-products of the fish-processing industry may cause environmental pollution. Sardinella gibbosa or goldstripe sardinella is commonly used in surimi and fish cracker productions in Malaysia. However, data on the nutritional composition of goldstripe sardinella fillets (GSF) and by-products (GSB) remain limited. The objectives of this study are to determine the nutritional composition and thermal properties of fillets and by-products of S. gibbosa. Results showed that the protein and moisture contents of GSF were higher than GSB. Conversely, the ash and fat levels of GSF were lower than GSB. Levels of saturated fatty acids (SFA) in GSF and GSB were higher than monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids. Palmitic acid and oleic acid were the major SFA and MUFA found in GSF and GSB. Both GSF and GSB contain higher percentages of DHA and EPA. Melting and crystallization profiles of the oil from S. gibbosa fillets were similar to by-product oil, which was related to their fatty acid content.     

Conjugated linoleic acid in diets for lambari (Astyanax altiparanae) (Garutti & Britski, 2000)

This study aimed to evaluate the effect of different levels of conjugated linoleic acid (CLA) on growth performance, carcass chemical composition and fatty acid profile of lambari (Astyanax altiparanae). A completely randomized experimental design with six treatments, diets with graded levels of CLA (0.0, 5.0, 10.0, 15.0, 20.0 and 25.0 g kg^?1) and five repetitions. Fish (n = 570; weight: 1.58 ± 0.23 g) were distributed into 30 (70‐L) aquaria and fed the experimental diets during 90 days. No effects of dietary supplementation with CLA on fish performance and carcass chemical composition were observed. CLA influenced carcass fatty acid profile, a positive relationship was found for 16:1 n‐9, 18:1 n‐9, 18:2(cis‐9, trans‐11), 18:2(trans‐10, cis‐12) and 20:1 n‐9, and a negative relationship was found for 15:1 n‐7, 16:0, 16:1 n‐5, 18:2 n‐6, 18:3 n‐3, 20:5 n‐3 and 22:4 n‐6. Total CLA and monounsaturated fatty acids (MUFA) linearly increased with the increase in dietary CLA, while saturated fatty acids (SFA) and polyunsaturated fatty acids (PUFA) of n‐6 and n‐3 series linearly decreased. Dietary CLA can be incorporated into the lambari (A. altiparanae) muscle, and the fish can be used as functional foods, because CLA is related to the prevention of various diseases in humans.     

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.     

Effect of combination of dietary Bacillus subtilis and trans‐cinnamic acid on innate immune responses and resistance of rainbow trout,Oncorhynchus mykiss to Yersinia ruckeri

The present study investigated the effects of combination of dietary Bacillus subtilis and trans‐cinnamic acid on serum biochemical parameters, innate immune responses and resistance of rainbow trout, Oncorhynchus mykiss to Yersinia ruckeri. Six experimental groups of fish with mean weights of 20.58 ± 0.35 g were used in the study. Five experimental groups of fish were fed diets containing Bacillus subtilis (10⁷ per gram) or a mix of the Bacillus subtilis (BS) and trans‐cinnamic acid (25 mg/kg‐25trcBS, 50 mg/kg‐50trcBS, 75 mg/kg‐75 trcBS, 150 mg/kg‐150 trcBS), whereas an additive‐free basal diet served as the control (Cont). In this study, an increase was observed in granulocyte percentage, respiratory burst activity, phagocytic activity, phagocytic index, myeloperoxidase activity and total antiprotease activity especially in fish fed with mix of the BS and trans‐cinnamic acid‐supplemented diets (p < .05). Moreover, at the end of the 20‐day challenge period the survival rates and antibody titre (p < .05), and relative per cent survival were higher in the BS group and all trcBS groups compared with control group. As a conclusion, the results in the present study show that feeding rainbow trout with diets containing a mix of B. subtilis and trans‐cinnamic acid over a 60‐day period might be sufficient for improving fish immune responses and disease resistance against Y. ruckeri.     

Differences in fatty acid composition of egg capsules from broodstock spotted babylon, <Emphasis Type="Italic">Babylonia areolata</Emphasis>, fed a local trash fish and formulated diet under hatchery conditions

This study is the first attempt to condition broodstock Babylonia areolata using formulated diets under hatchery conditions. Samples of spotted babylon egg capsules from broodstock fed either a formulated diet or a local trash fish, carangid fish (Seleroides leptolepis) for 120 days were analyzed for proximate composition and fatty acid composition. The formulated diet contained significantly higher levels of arachidonic acid (20:4n − 6; ARA), eicosapentaenoic acid (20:5n − 3; EPA) and docosahexaenoic acid (22:6n − 3; DHA) than those of the local trash fish. The formulated diet also had significantly higher ratios of DHA/EPA and (n − 3)/(n − 6) PUFA than those of local trash fish but not for the ARA/EPA ratio. The compositions of egg capsules produced from broodstock fed formulated diet contained significantly more ARA, EPA and DHA compared to broodstock fed the local trash fish. The ARA/EPA and DHA/EPA ratios in egg capsules were significantly higher in the trash fish—fed group compared to those fed the formulated diet. However, (n − 3)/(n − 6) PUFA ratios in egg capsules produced from broodstock fed the formulated diet did not differ significantly compared to those from broodstock fed the local trash fish. The relatively low DHA/EPA, ARA/EPA and (n − 3)/(n − 6) ratios in the egg capsules produced from the formulated diet—fed broodstock B. areolata suggested that this diet is inferior, when compared to the traditional food of trash fish.     

Changes in the Quality of Refined Fish Oil in an Accelerated Storage Study

ABSTRACT

The aim of this study was to prepare the refined oil derived from Sardinella longiceps by a step-wise procedure of degumming, bleaching, and deodorization. The refined sardine oil showed greater susceptibility toward oxidation than the crude oil, but the addition of ethyl acetate fractions of the red seaweeds Hypnea musciformis, Kappaphycus alvarezii, and Jania rubens with great antioxidant potential significantly increased the oxidative stability of the refined oil. The refined oil supplemented with the ethyl acetate fraction of these seaweeds indicated more noteworthy oxidative stability indices (≥ 0. 51 h) than α-tocopherol. The induction time was greater for refined oil added with H. musciformis (1.26 h) than butylated hydroxytoluene (1.04 h) and α-tocopherol (0.4 h). No traces of aromatic (δ 6.7–7.5 ppm), hydroperoxides, and aldehydes (δ 9–10 ppm) in the ¹H-NMR spectra of the refined oil supplemented with seaweed extracts at the baseline and after the accelerated storage study demonstrated that the seaweed extracts were able to prevent the formation of secondary oxidation products in the unsaturated system during storage. This study demonstrated the potential of seaweeds as natural alternatives to the synthetic antioxidants to prevent the rancidity of the refined fish for use in food and pharmaceutical industries.     

Rhodovulum sulfidophilum,a phototrophic bacterium,grown in palm oil mill effluent improves the larval survival of marble goby Oxyeleotris marmorata (Bleeker)

Survival of marble goby larvae fed either Rhodovulum sulfidophilum, a phototrophic bacterium cultured from palm oil mill effluent (pPB), or microalgae ( Nannochloropsis sp.) was evaluated at two salinities. Larvae directly fed pPB had survival of 0–29% at 5 g L^?1 salinity and 0–19% at 10 g L^?1 salinity, whereas larvae directly fed microalgae suffered complete mortality after 20 days of culture at both salinities. However, larvae indirectly fed pPB or microalgae, i.e. via rotifers (Days 1–30) and Artemia nauplii (Days 21–30) cultured solely from pPB or microalgae, showed improved survival of 35–55% or 44–49% at 5 g L^?1 salinity respectively. In all experiments, fish larvae reared at 5 g L^?1 salinity showed significantly higher (P < 0.01) mean survival than those reared at 10 g L^?1 salinity. The survival of larvae fed the bacterial‐based diet was higher compared with microalgal diet used in previous studies. The pPB had higher total polyunsaturated fatty acids and docosahexaenoic acid (DHA) than the microalgae, which had very high eicosapentaenoic acid (EPA). Larvae with very high ratios of DHA/EPA (>11) or/and ARA (arachidonic acid)/EPA (>5), attributable to their given diet, however suffered the highest mortality.     

Sex organ extracts and artificial hormonal compounds as sex pheromones to attract broodfish and to induce spawning of Chinese black sleeper (Bostrichthys sinensis Lacépède)

Ovarian, testicular and seminal vesicle extracts of mature Chinese black sleeper (Bostrichthys sinensis Lacepede), synthetic steroids 17α‐progesterone (17α‐P), 17α,20β‐dihydroxy‐4‐pregnen‐3one (17α,20β‐P) and prostaglandins PGE₂ and PGF_2α were used in an attempt to attract broodfish and to induce spawning. Furthermore, 17α‐P, PGE₂ and PGF_2α levels were determined in extracts, and PGE₂ and PGF_2α levels were determined in holding and spawning waters. The results showed that ovarian extracts attracted more males than females to enter artificial nests, while testicular and seminal vesicle extracts attracted more females than males. The highest percentage of spawned nests were found when 17α,20β‐P and PGE₂ were administered to the nests, while the largest egg numbers and the mean highest fertilization rate were observed when PGE₂ was added. In ovarian extract, 17α‐P level was higher than levels in testicular or seminal vesicle extracts. The PGE₂ levels were significantly (P<0.05) higher than PGF_2α in the ovarian, testicular and seminal vesicle extracts. The PGE₂ and PGF_2α levels in spawning waters were higher than those in holding waters. The PGE₂ levels were greatly significantly (P<0.01) higher than PGF_2α in spawning and in holding waters. The results suggest that the sex organs of B. sinensis contain 17α‐P, 17α, 20β‐P, PGE₂ and PGF_2α and that 17α‐P, 17α,20β‐P and PGE₂ may act as sex pheromones in this species, attracting both male and female conspesifics to spawning sites and inducing spawning. Among the compounds tested, PGE₂ was the most effective sex pheromone for induction of spawning in B. sinensis.     

No significant effect of additive ratios of docosahexaenoic acid to eicosapentaenoic acid on the survival and growth of cobia (Rachycentron canadum) juvenile

An experiment was conducted in the laboratory to investigate the effects of additive ratios of docosahexaenoic acid (DHA) to eicosapentaenoic acid (EPA) on the growth and survival of cobia (Rachycentron canadum) juveniles from August to October 2005. Three hundred and eighty cobia juveniles (56 days of age, body weight 6.9 ± 0.1 g, body length 9.2 ± 0.1 cm) were selected and 20 of them were freely taken for initial sample analysis in the week 0. Additional 360 juveniles were randomly assigned into eight groups with triplicate, total 24 tanks with 15 fish each. Cobia juveniles were reared in glass‐steel tanks (200‐L volume per tank) using filtered seawater with temperature 26–30.5 °C, salinity 25.4–33.0 g L^?1 and pH 7.8–8.0. Cobia juveniles were fed for 8 weeks using seven treatment diets (D‐1 to D‐7) with the same amount of DHA and EPA (15.0 ± 1.2 g kg^?1 of dried diet), but varying ratios of DHA to EPA (0.9, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, respectively) and a control diet (D‐0, DHA + EPA = 8.0 g kg^?1 of dried diet, DHA/EPA = 1.3). Five juveniles per tank were randomly taken for sample analysis at the end of weeks 4 and 8, respectively. The highest protein efficiency rate (PER; 1.5 in mean), average body weight (BW; 73.3 g per fish in mean) and the lowest feed conversion ratio (FCR; 1.6 in mean) were obtained in cobia juveniles fed the control diet at the end of week 8. These parameters were significantly different (P < 0.05) among juveniles fed the control and treatment diets; however, no significant difference (P > 0.05) was found among juveniles fed the treatment diets evaluated in this study. It was concluded that the survival and growth of cobia juveniles were not greatly influenced by additive ratios of DHA to EPA in our experimental conditions.     

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.     

Effects of dietary fatty acids on the production and quality of eggs and larvae of Atlantic cod (Gadus morhua L.)

Cultivated Atlantic cod (Gadus morhua) entering their first year of gamete maturation were fed diets with different levels of arachidonic acid (ARA) and eicosapentaenoic acid (EPA) for 6.5 months prior to commencement of spawning. Gravid females were stripped three times: at the beginning, peak and end of spawning. Lipid composition and egg and larval quality of 34 family crosses were investigated. Results indicated that ARA uptake into eggs from broodstock diet was highly efficient achieving proportions of ARA up to 84% higher in eggs than in the diet. EPA was 42–76% higher, and DHA was 155–173% higher in eggs than in diets. Cod fed the diet with the lowest EPA/ARA ratio had the greatest egg production. Eggs from fish on a diet with high ARA level had significantly higher fertilization and hatching success than those fed low levels of ARA. This diet produced on average 71 viable eggs g^?1 female compared with 32.5 and 4 eggs in diet B and C, respectively. Furthermore, larval survival until 8 days posthatch was higher in diets with lower ARA levels. The combined results showed that ARA dietary supplementation and low EPA/ARA ratio yielded a greater number of viable larvae kg^?1 female.     

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.     

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.