Predator-prey interactions between Isochrysis galbana and Oxyrrhis marina. II. Release of non-protein amines and faeces during predation of Isochrysis

During the growth of Isochrysis galbana, several non-proteinamines may be detected in the growth medium. Of these, one (termed‘TTl’) accumulates in proportion to the numbersof cells present. The concentrations of ‘TTl’, andof another (termed ‘TA’), are 3–5 times higherin cultures in which Isochrysis is predated by Oxyrrhis marina.The lowest estimates of the concentration of extracellular ‘TTl’are an order of magnitude higher than those of any protein aminoacid. Of the protein amino acids, some like glycine are utilizedduring predation while others, like histidine, accumulate inthe medium Because of the unknown N-content and reactivity ofthe non-protein amines during HPLC, it is not possible to sayif these compounds (together with other components of dissolvedorganic N) form a significant proportion of the unaccountedfor N in the system after predatory activity. During predationin the absence of detectable free ammonium (when Isochrysismay be expected to be N-deprived), particles accumulate in themedium. Most of these are <2.5 µ.m in diameter andare suggested to be remains of digested prey. There is evidenceof a reassimilation of these particles by prey-deplete Oxyrrhis.     

Growth and pigment accumulation in nutrient-depleted Isochrysis aff. galbana T-ISO

The effect of three different nutrient depletions (nitrogen, sulphur and magnesium) on the growth and pigment accumulation of the haptophyte Isochrysis aff. galbana (clone T-ISO) has been studied. Pigments were quantified based on RP-UHPLC-PDA-MSⁿ analysis. All nutrient depletions led to reduced maximal biomass concentrations. Besides, all nutrient-depleted cultures accumulated 3-hydroxyechinenone. To our knowledge, this is the first time that 3-hydroxyechinenone has been found in I. aff. galbana T-ISO. Most 3-hydroxyechinenone, as well as the most echinenone and diatoxanthin, were found in the nitrogen-limited culture in which a more severe limitation resulted in higher cellular contents. Similar to accumulation of diatoxanthin, accumulation of 3-hydroxyechinenone and echinenone may be part of a global (stress) response mechanism to oversaturating light conditions.     

There are high levels of functional and genetic diversity in Oxyrrhis marina

Oxyrrhis marina, a widely distributed marine protist, is used to model heterotrophic flagellate responses in microbial food webs. Although clonal variability occurs in protists, assessments of intraspecific diversity are rare; such assessments are critical, particularly where species are used as models in ecological studies. To address the extent of intraspecific variation within O. marina, we assessed diversity among 11 strains using 5.8S rDNA and ITS sequences. The 5.8S rDNA and ITS regions revealed high divergence between strains: 63.1% between the most diverse. To compare O. marina diversity relative to other alveolates, 18S rDNA sequences for five strains were analysed with sequences from representatives of the major alveolate groups. 18S rDNA also revealed high divergence in O. marina. Additionally, consistent with phylogenies based on protein coding genes, maximum likelihood analysis indicated that O. marina was monophyletic and ancestral to the dinoflagellates. To assess ecophysiological differences, growth rates of seven O. marina strains were measured at 10 salinities (10-55 per thousand). Two salinity responses occurred: one group achieved highest growth rates at high salinities; the other grew best at low salinities. There was no clear correlation between molecular, ecophysiological, or geographical differences. However, salinity tolerance was associated with habitat type: intertidal strains grew best at high salinities; open-water strains grew best at low salinities. These data indicate the need to examine many strains of a species in both phylogenetic and ecological studies, especially where key-species are used to model ecological processes.     

Predator-prey interactions: effects of carp predation on Tubificid dynamics and carp production in experimental fishpond

Effects of predation by carp (Cyprinus carpio L) were analysed in an experimental fishpond where the main components of the benthic fauna were tubificids. Enclosures inaccessible to fishes were compared with the rest of the pond stocked with 30 young carps (1⁺). Fish predation reduced the density, biomass and production of prey; the production of Tubificidae inside the enclosures was 1.7 times higher than that of the part of the pond stocked with fishes. On the other hand tubificid turnover ratios (P/B) inside and outside enclosures were not significantly different (respectively 5.57 and 5.38), and the size distribution of tubificids was not significantly modified. The ratio of the energetic equivalent of net fish production to biomass of tubificids consumed was 5.59%.     

NaNO3浓度对球等鞭金藻生长及所含脂肪酸的影响

用NaNO3作氮源,分别作了氮浓度的5个水平对球等鞭金藻H29的相对生长速率、总脂含量及脂肪酸组成的影响实验。实验结果显示,H29的相对生长率和总脂含量随N浓度的增加而增加,在897.6 mg/L时总脂含量达到最大值(23.4%)。而DHA(22:6n-3)和PUFAs(polyunsaturated fatty acids)含量随氮浓度的改变有较大变化,DHA含量总体随N浓度增加而下降,在较低氮浓度(74.8mg/L)时含量达到最大值12.11%。     

Light and prey influence the abundances of two rhodopsins in the dinoflagellate Oxyrrhis marina

Protoplasma - Antisera were raised against the C-terminal amino acid sequences of the two rhodopsins ADY17806 and AEA49880 of Oxyrrhis marina. The antisera and affinity-purified antibodies thereof...     

Cellular accumulation and distribution of cadmium in Isochrysis galbana during growth inhibition and recovery

Population growth of Isochrysis galbana in batch cultures containingcadmium in solution (1 and 100 µg Cd 1^–1) recoveredafter an initial period of inhibition. This recovery did notappear to be due either to a loss of Cd from the culture orto the development of a Cd exclusion mechanism by cells. Gelfiltration of cell cytosol revealed the association of Cd withfour chromatographically distinct fractions. The acquisitionof tolerance to Cd by I. galbana appeared to be correlated withchanges in the cytosolic distribution of Cd, including an increasedbinding of Cd to a small molecular weight cytosolic moiety.     

NaNO浓度对球等鞭金藻生长及所含脂肪酸的影响

用NANO3作氮源,分别作了氮浓度的5个水平对球等鞭金藻H29的相对生长速率、总脂含量及脂肪酸组成的影响实验．实验结果显示,H29的相对生长率和总脂含量随N浓度的增加而增加,在897．6mg/L时总脂含量达到最关值（23.4％）：而DHA（22：6n-3）和PUFAs（polyunsaturated fatty acids）含量随氮浓度的改变有较大变化,DHA含量总体随N浓度增加而下降,在较低氮浓度（74．8mg／L）时含量达到最大值12．11％。     

Effects of temperature and growth phase on lipid and biochemical composition of Isochrysis galbana TK1

The lipid and biochemical composition of the haptophyte Isochrysis galbana TK1 was examined. Cultures were grown at 15 °C and 30 °C, and harvested in the exponential and early stationary growth phases. Carbohydrate and protein content varied at the two culture temperatures and growth phases. The highest protein content was found at the exponential growth phase at 15 °C, and the highest carbohydrate content was found at the stationary phase at the same culture temperature. Lipid accumulated in the stationary growth phase and its content was higher at 30 °C than at 15 °C regardless of the growth phase. The neutral lipids were the major class of lipid found in all the cultures. The stationary phase culture had a higher proportion of neutral lipids than the exponential phase culture and the proportion decreased slightly when culture temperature was increased from 15 °C to 30 °C. Phospholipid levels remained constant at the two temperatures, but slightly decreased in the stationary phase. Glycolipids in the exponentially growing cells were higher than those from stationary growth phase and increased with temperature. Polyunsaturated fatty acids (PUFAs) predominated in glycolipids and phospholipids. Cells grown at 15 °C contained higher proportion of 18:3 (n–3) and 22:6 (n–3) with a corresponding decrease in 18:2 (n–6), monounsaturated and saturated fatty acids. This revised version was published online in August 2006 with corrections to the Cover Date.     

Mathematical modelling of extracellular matrix dynamics using discrete cells: fiber orientation and tissue regeneration.

Matrix orientation plays a crucial role in determining the severity of scar tissue after dermal wounding. We present a model framework which allows us to examine the interaction of many of the factors involved in orientation and alignment. Within this framework, cells are considered as discrete objects, while the matrix is modelled as a continuum. Using numerical simulations, we investigate the effect on alignment of changing cell properties and of varying cell interactions with collagen and fibrin.     

Sustainable, High-Yielding Outdoor Mass Cultures of Chaetoceros muelleri var. subsalsum and Isochrysis galbana in Vertical Plate Reactors

Continuous cultures of Chaetoceros muelleri and Isochrysis galbana were grown outdoors in flat plate-glass reactors in which light-path length (LPL) varied from 5 to 30 cm. High daily productivity (13 to 16 g cell mass per square meter of irradiated reactor surface) for long periods of time was obtained in reactors in which the optical path as well as cell density were optimized. 'Twenty centimeters was the optimal LPL, yielding the highest areal productivity of cell mass (g m^–2d^–1), eicosapentaenoic acid, and docosahexaenoic acid, which was identical with that previously found for polysaccharide production of Porphyridium and not far from the optimal LPL affecting maximal productivity in Nannochloropsis species. Relating the energy impinging on a given reactor surface area to the appropriate number of cells showed that the most efficient light dose per cell, obtained with the 20-cm LPL reactor, was approximately 2.5 times lower than the light dose available per cell in the 5-cm LPL reactor, in which a significant decline in areal cell density accompanied the lowest areal output of cell mass. The most effective harvesting regimen was in the range of 10% to 15% of culture volume harvested daily and replaced with fresh growth medium, resulting in a sustainable culture density of 24 × 10⁶ and 28 × 10⁶ cells/ml of C. muelleri and I. galbana, respectively.     

Biomass production and biochemical variability of the marine microalga Isochrysis galbana in relation to culture medium

We have studied the autotrophic growth of the marine microalga, Isochrysis galbana Parke, in a batch photobioreactor, comparing five different culture media and analysing the influence of each on growth kinetics as well as on the fatty-acid composition and protein content of the biomass. All the experiments were performed at 15 degrees C, with the culture medium at pH 8.0, a specific rate of air supply of 1vv(-1)min(-1) and a continuous illumination of 40-43Wm(-2). The results show no parallel between good nutritional characteristics and high values of the kinetic parameters. Nevertheless, a compromise between the nutritional factors and growth kinetics could be provided by Ukeles medium, which provided a biomass with a good composition in polyunsaturated fatty acids (quotient n3/n6=3.2), an adequate protein content (25.3%) and relatively high values, although not the highest registered, for maximum specific growth rate (μ(m)=0.018h(-1)) and biomass productivity (1.9x10(-3)kgm(-3)h(-1)).     

Cloning and expression of a gene from Isochrysis galbana modifying fatty acid profiles in Escherichia coli

An ester hydrolase gene from the microalga Isochrysis galbana was cloned and expressed in Escherichia coli BL21 Rosetta 2?. The full-length putative gene has 1,146 base pairs and codes for a 381-amino acid polypeptide. The predicted molecular mass of the deduced protein is approximately 42.31 kDa, with a theoretical pI of 9.37. Slight similarity and identity were observed between the microalga sequence and various α/β-fold hydrolases found in diverse phyla. The catalytic triad corresponds to residues Ser²⁵⁴, Asp³⁰⁹, and His³⁴¹, with the nucleophilic catalytic residue Ser²⁵⁴ located in the pentapeptide consensus motif G-X-S²⁵⁴-X-G. The activity of the enzyme was established by fatty acid profile analysis of the membrane lipids. The expression of the protein in E. coli shifted the fatty acid composition predominantly towards C16:1 and C18:1 fatty acids. This enzyme is called I. galbana thioesterase/carboxylesterase (or IgTeCe). This novel gene is shown to have a potential for use in metabolic engineering to enhance the lipid yields of microalgae.