Jose R. Rainuzzo

The significance of lipids at early stages of marine fish: a review

The present work reviews the significance of lipids at different early stages of marine fish larvae. Lipids in broodstock nutrition are considered to be important for the quality of the larvae. Lipids affect the spawning and the egg quality of many fish species and a deficiency in (n−3) highly unsaturated fatty acids (HUFA) in broodstock negatively affects fecundity, fertilization rate and hatching rate of the species studied. Lipids as a source of energy at the embryonic and larval stage (before first-feeding) are evaluated in relation to other sources of energy such as protein and carbohydrates. After hatching and prior to first-feeding, some marine species show a preference in catabolizing phosphatidylcholine, whereas phosphatidylethanolamine tends to be synthesized. The effect of long-term (LT) and short-term (ST) enrichment techniques on the lipid composition of rotifers has been documented using various marine oils/emulsions. The quantitative and qualitative lipid class and fatty acid composition of diets influenced the lipid and fatty acid composition of both LT- or ST-enriched rotifers. The nutritional improvement of Artemia is also important and may follow the general methods used for rotifers. The functions of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) during early stages of marine fish larvae are apparently different. High amounts of EPA in relation to DHA may create an imbalance in the structural composition of the phospholipids, which could affect the normal growth and the quality of the larvae. Turbot larvae tended to exhibit lower pigmentation success with lower DHA:EPA ratio in the total lipid fraction of the larvae, especially when the absolute amounts of EPA were high compared to those of DHA (in the total lipid and phospholipid fraction of the larvae). Considerable research is necessary to clarify many aspects regarding the function of these fatty acids, especially how their content at the egg stage can affect further requirements for normal growth and survival.

EFFECT OF NUTRIENT LIMITATION ON FATTY ACID AND LIPID CONTENT OF MARINE MICROALGAE1

Phaeodactylum tricornutum and Chaetoceros sp. (Badllariophyceae), Isochrysis galbana (clone T‐Iso) and Pavlova lutheri (Prymnesiophyceae), Nannochloris atomus (Chlorophyceae), Tetraselmis sp. (Prasinophyceae), and Gymnodinum sp. (Dinophyceae) were cultured at different extents of nutrient‐limited growth: 50 and 5% of μmax. The lipid content of the algae was in the range 8.3–29.5% of dry matter and was generally higher in the Prymnesiophyceae than in the Prasinophyceae and the Chlorophyceae. Increasing extent of phosphorus limitation resulted in increased lipid content in the Bacillariophyceae and Prymnesiophyceae and decreased lipid content in the green flagellates N. atomus and Tetraselmis sp.

A review of the nutritional effects of algae in marine fish larvae

In the first-feeding of larval, turbot (Scophthalmus maximus) and halibut (Hippoglossus hippoglossus), microalgae are used in the production of rotifers (Brachionus plicatilis) in order to transfer essential nutrients from the algae to the live food. In addition, the algae may be given directly to the larvae along with the live food. In this circumstance they act both as food for the fish larvae and for the live food. Microalgal addition to the first-feeding tanks along with the rotifers improved growth and survival of larvae, whereas short-term enrichment of rotifers with algae did not improve growth and survival of larvae in tanks without algae added. The algae in larval tanks tended to modify and stabilize the nutritional quality of the rotifers in the period before they were consumed by the larvae. The lipid content and fatty acid composition of the rotifers reflected the composition of the algal diets, and the algal species used may be an effective tool to control the fatty acid content (especially DHA, 22:6n−3, docosahexaenoic acid, and EPA, 20:5n−3, eicosapentaenoic acid). The content of n−3 polyunsaturated fatty acids varied between algal species, and to some extent, with the growth limitation of the algal cells. Young stages of marine fish larvae ingested microalgae, but the algal cells were assimilated to a different extent in halibut and turbot. The ingested microalgae may have triggered the digestion process or contributed to the establishment of an early gut flora. The algae in larval tanks most probably modified the bacterial flora of the water and the rotifers. In addition, the algae in larval tanks may have modified the light milieu for the larvae.

Nutritional effects of algal addition in first-feeding of turbot (Scophthalmus maximus L.) larvae

Abstract The effect which microalgal addition and rotifer enrichment with algae had on survival, growth rate and fry viability during first-feeding were examined for turbot (Scophthalmus maximus L.). Addition of the microalgae Isochrysis galbana or Tetraselmis sp. together with rotifers Brachionus plicatilis, grown on yeast and oil emulsion, and Artemia greatly improved rearing success, whereas short-term enrichment of the rotifers with Tetraselmis sp. gave only improved viability. The algae modified the relative fatty acid composition of rotifers. Rotifers with I. galbana exhibited and increased level of 22:6n-3 and a lower level of 20:5n-3, whereas the opposite change occurred when Tetraselmis sp. was used. Addition of either of the two algal species at 1 mg cl−1 to the larval tanks resulted in a constant lipid level and high egg ratio of the rotifers, and thereby also high individual biomass content. Without algal addition, the lipid content of the rotifers decreased by 20% day−1 in the early phase, and their egg ratio became close to zero. In addition to improved nutritional conditions of the larvae, some other factor, e.g. some trigger mechanism or changed microbial or light conditions, also can operate to explain the increased early appetite of larvae with microalgae added and must be taken into consideration.

Influence of lipid composition of live feed on growth, survival and pigmentation of turbot larvae

The effect of different lipid compositions of live feed on the survival, growth rate and pigmentation success of turbot larvae, Scophthalmus maximus (L.), was investigated. Rotifers, Brachionus plicatilis, together with the algae Tetraselmis sp., were administered until day 12, and Artemia was fed until day 27. The experimentally treated live feeds were enriched with four formulated emulsions, resulting in a gradient in the relative contents of Ω3 HUFA (highly unsaturated fatty acids) and in DHA (docosahexaenoic acid, 22:6 Ω3)/EPA (eicosapentaenoic acid, 20:5 Ω3) ratios in both the rotifers and Artemia.There were no differences in larval growth rate, and only small differences in survival rate throughout the feeding experiment, probably because of satisfactory levels of Ω3 HUFA in the live feed to sustain growth and survival. A correlation was obtained between the percentage of completely pigmented 27 d old turbot and the DHA/EPA ratio in the total lipids of 12 d old larvae, which again was correlated with the corresponding ratio in the live feed used. The results suggest that normal pigmentation in turbot requires dietary DHA in the early larval feeding period, and that this requirement cannot be replaced by EPA.

Comparative study on the fatty acid and lipid composition of four marine fish larvae

Abstract 1. 1. Fatty acid and lipid class composition were determined in larvae of four marine species: Atlantic halibut (Hippoglossus hippoglossus L.), plaice (Pleuronectes platessa), cod (Gadus morhua) and turbot (Scophthalmus maximus) at hatching and prior to first feeding. 2. 2. Total fatty acid content decreased in the four species with up to 50% reduction in one of the halibut groups. Docosahexanaoic acid (22:6 n-3) was especially utilized. 3. 3. Low lipid utilization was found in turbot in relation to the other three species. 4. 4. Water environmental temperature may explain some of the differences in the fatty acid utilization and the source of metabolic energy between cold water species (halibut, cod, and plaice) and temperate species (turbot), in the period from hatching to prior to first feeding. 5. 5. Relative amounts of neutral lipids and phospholipids were similar in plaice, cod and halibut, approximately 25% and 75% of total lipids, respectively, and were approximately constant during the yolk-sac stage. Neutral lipids were dominant for turbot at hatching, accounting for 53–55% of the total lipids, while phospholipids predominated prior to first feeding, being 56–59%. 6. 6. Phosphatidylcholine was catabolized in halibut, plaice and cod but not in turbot, while phosphatidylethanolamine tended to be synthesized in all four species.

Lipid composition in turbot larvae fed live feed cultured by emulsions of different lipid classes

Fatty acid and lipid class compositions were determined in eggs and larvae of turbot (Scophthalmus maximus L.). The larvae were fed on rotifers and Anemia fed on various lipid emulsions. Starving larvae were also studied. The lipid class compositions of the emulsions affect the fatty acid composition of the rotifers. Ethyl ester-based emulsion exhibited the highest assimilation by the rotifers and Artemia and were possibly incorporated in their triacylglycerol fraction. During larval starvation docosahexaenoic acid and arachidonic acid levels tended to be conserved whereas eicosapentaenoic acid was highly reduced. Moreover, the relative proportion of phosphatidylethanolamine increased whereas that of phosphatidylcholine decreased in starved larvae. Increasing the n-3 highly unsaturated fatty acid levels in rotifers and Artemia did not influence the survival and growth of the turbot larvae. A positive correlation was found between pigmentation success and the ratios of docosahexaenoic/ eicosapentaenoic acids in the total and polar lipid fractions of the turbot larvae. Furthermore, the amounts of these polyunsaturated fatty acids in the phosphatidylethanolamine fraction of the larvae are suggested to be of particular importance in turbot pigmentation.

Effect of short- and long-term lipid enrichment on total lipids, lipid class and fatty acid composition in rotifers

Four emulsions differing in lipid class composition: triacylglycerols, ethyl esters, phospholipids and wax esters were used to enrich rotifers either through short-term (ST) enrichment (24 h) or through long-term (LT) enrichment (10 days). Higher lipid levels were obtained by using the ST enrichment method. This was particularly marked in the high triacylglycerol accumulation in rotifers enriched on the phospholipid-based emulsion. Ethyl esters were effectively assimilated and incorporated into triacylglycerol by rotifers in both the ST and LT techniques. A high docosaehexanoic/eicosapentaenoic acid (DHA/EPA) ratio was obtained in the LT technique using the ethyl ester-based emulsion. However, the other emulsion treatments gave higher or equal DHA/ EPA ratios using the ST technique. Absolute phospholipid levels were independent of both dietary lipid composition and enrichment method used, whereas triacylglycerol levels depended on these parameters. During starvation the level of phospholipid, in absolute terms, decreased slightly whereas the triacylglycerol fraction decreased considerably. Rotifers enriched on the wax ester-based emulsion using the LT technique exhibited higher levels of long-chain monoenes (i.e. 20:1 and 22:1 isomers) than when the ST technique was used. This suggests that hydrolysis of the wax esters and oxidation of the liberated fatty alcohols to fatty acids occurred when the LT technique was applied.

Quantitative high-resolution13C and1H nuclear magnetic resonance of ω3 fatty acids from white muscle of atlantic salmon (Salmo salar)

High-resolution13C nuclear magnetic resonance (NMR) spectra have been obtained and used to define the ω3 (n-3) fatty acid distribution in lipid extract and white muscle from Atlantic salmon (Salmo salar). The13C spectrum of lipid extracted from muscle gives quantitative information about the individual n-3 fatty acids, 18:2n-6, 20:1/22:1 and groups of fatty acids. The quantitative data compare favorably with those obtained by gas-liquid chromatography. The1H NMR spectrum of the lipid extract gives information about the amount of 22:6n-3 and the total content of n-3 fatty acids. The13C NMR technique also revealed the positional distribution (1,3- and 2-acyl) of the important 20:5n-3 and 22:6n-3 acids in the triacylglycerol molecules. In the quantitative13C NMR spectrum of white muscle, the methyl region of the acyl chains of triacylglycerols gave rise to sufficiently resolved signals to permit estimation of the total concentration of lipids and the n-3 fatty acid content. The NMR data are in good agreement with corresponding data obtained by traditional methods.

Acclimation of Atlantic cod (Gadus morhua) to cold water: Sxtress response, osmoregulation, gill lipid composition and gill Na-K-ATPase activity

Abstract Atlantic cod (0.8–2.5 kg) were transferred froni 8°C to 1°C seawater for 17 days. No fish died. Exposure to 1°C water produced no changes in hematocrit, or in plasma concentrations of Cl − or Mg 2+ . Cold water exposure caused a marked increase in plasma cortisol and glucose concentrations. Fish in 1°C seawater had higher gill Na-K-ATPase activity than fish in 8°C seawater, whereas there were no differences in gill lipid class or fatty acid composition.