Yngvar Olsen

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.

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.

Microbially matured water: a technique for selection of a non-opportunistic bacterial flora in water that may improve performance of marine larvae

Before transfer to larval incubators, water was membrane filtered to remove >95% of the bacteria and then transiently maintained in a biofilter that promoted recolonization of the water by non-opportunistic bacteria. The process is termed microbial maturation of the water. Hypothetically the bacterial flora in the matured water should protect the marine larvae from colonization and proliferation by opportunistic bacteria. Testing of the hypothesis demonstrated 76% higher survival of yolk sac larvae of Atlantic halibut (Hippoglossus hippoglossus) in matured than in membrane filtered water. Proliferation of opportunistic bacteria was observed in the rearing water after hatching of turbot eggs (Scophthalmus maximus), but to a less extent in the microbially matured water. In the early phase of first feeding of turbot larvae, the matured water induced qualitative differences in the gut microflora. Significantly higher initial growth rate of the turbot larvae in the matured water affected 51% higher average weight of 13 days old larvae than in membrane filtered water. Algal addition to the matured water enhanced the larval growth further. The experiments conducted supported the proposed hypothesis that microbial maturation selects for non-opportunistic bacteria, which protects the marine larvae from proliferation of detrimental opportunistic bacteria.

Protein and carbon utilization of rotifers (Brachionus plicatilis) in first feeding of turbot larvae (Scophthalmus maximus L.)

Abstract The effect of three different rotifer enrichments was examined on growth, survival, pigmentation and viability of first feeding turbot larvae. The diets differed in rotifer content of protein, lipid and ratio of protein/lipid. The diets were fed to turbot with or without algae ( Isochrysis galbana ) added to the larval tanks. The turbot larvae were fed rotifers for 10 days and thereafter the same Artemia diet was fed to all treatments for the rest of the experimental period. Growth and survival of fish larvae were higher in tanks containing algae than in tanks where no algae were added. Independent of algal addition, the highest growth rate and survival was obtained by feeding rotifers containing the highest protein content. Larvae reared in greenwater consumed higher numbers of rotifers during the stagnant period than larvae kept in clearwater conditions, while analysis of the larval gut contents showed lower rotifer numbers in the gut of larvae reared in greenwater conditions. This must imply longer residence time of the food in the larval gut, and presumably also higher digestion and assimilation efficiencies of larvae maintained without algae than in larvae maintained with algae. Calculation of protein and carbon conversion efficiency showed higher utilization in larvae maintained without algae (18–28% for protein, 12–19% for carbon) than in larvae maintained with algae (6–9% for protein, 4–7% for carbon). No significant differences in pigmentation rate and stress sensitivity were observed among the larvae of the various treatments.

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.

Dependence of temperature on loss rates of rotifers, lipids, and ω3 fatty acids in starved Brachionus plicatilis cultures

Rotifer cultures of Brachionus plicatilis (SINTEF-strain, length 250 μm) rich in ω3 fatty acids were starved for > 5 days at variable temperature (0–18 °C). The net specific loss rate of rotifer numbers were 0.04 day−1 (range 0–0.08 day−1) at 5–18 °C, but reached values up to 0.25 day−1 at 0–3 °C. The loss rate was independent on culture density (range 40–1000 ind ml−1), but was to some extent dependent on the initial physiological state of the rotifers (i.e., egg ratio).

Particle size dependent feeding by the rotifer Brachionus plicatilis

AbstractSize selective feeding by Brachionus plicatilis was investigated with algae and bacteria (0.3–3.5 µm) and mono-disperse latex beads (0.3–3.0 µm) in short term feeding experiments. B. plicatilis demonstrated maximum clearance rate of particles with diameter ≥2µm, but particles with diameter down to 0.3 µm were also ingested. The clearance rate of bacteria was 15–55% of that obtained for optimal sized particles (≥2 µm), and was related to particle size. The relative reduction in retention of particles with diameter < 2 µm was more pronounced for latex beads than for natural food particles, suggesting other mechanisms than size to be important for the particle retention by the rotifer. This is emphasized by the fact that the clearance rates were much lower for latex beads than for natural food particles of comparable size. Efficient retention of bacteria was observed for rotifers in poor physiological condition, i.e. rotifers with low maximum clearance rate. This may reflect a strategy to optimize energy utilization by reducing locomotion costs and increasing energy intake.The results indicate that B. plicatilis has a low to medium ability to feed on bacteria. In natural ecosystems, its importance as a bacterial grazer is of limited importance. At high population densities, such as in live feed cultures, the rotifer may, however, efficiently remove bacteria from the culture.

Evaluation of competitive ability of Staurastrum luetkemuellerii (Chlorophyceae) and Microcystis aeruginosa (Cyanophyceae) under P limitation

The desmid Staurastrum luetkemuellerii Donat et Ruttner and the cyanobacterium Microcystis aeruginosa Kütz. showed pronounced differences in chemical composition and ability to maintain P fluxes. The cellular P:C ratio (Qp) and the surplus P:C ratio (Qsp) were higher in M. aeruginosa, indicating a lower yield of biomass C per unit of P. The subsistence quota (Qp) was 1.85 μg P·mg C−1in S. luetkemuellerii and 6.09 μg P·mg C−1in M. aeruginosa, whereas the respective Qp of P saturnted organisms (Qs) were 43 and 63 μg P·mg C−1. These stores could support four divisions in S. luetkemuellerii and three divisions in M. aeruginosa, which suggests that the former exhibited highest storage capacity (Qs/Q0). M. aeruginosa showed a tenfold higher activity of alkaline phosphatase than S. luetkemuellerii when P starved. The optimum N:P ratio (by weight) was 5 in S. luetkemuellerii and 7 in M. aeruginosa.