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Biomedical subjects

J Abalde

Publications and source records attributed to J Abalde.

6 recordsLinked to original sources

Analysis and enhancement of astaxanthin accumulation in Haematococcus pluvialis.

The green microalga Haematococcus pluvialis was cultured with different concentrations of NaNO(3) to determine the effect on cell growth and astaxanthin accumulation. The optimum nitrate concentration to obtain astaxanthin and to avoid the cessation of cell division was 0.15 g/l NaNO(3). The ratio chlorophyll a/total carotenoids proved a good physiological indicator of nitrogen deficiency in the cell. The effect of different carbon sources, malonate and acetate, on astaxanthin accumulation was also studied; up to 13 times more carotenoids per cell were accumulated in cultures with malonate than in cultures without this compound. The pigment analysis was performed by a new low toxicity HPLC method capable of separating chlorophylls a and b, carotenes and xanthophylls in a short-period of time, using low volumes of solvents and with an economical price. With this method even echinenone was separated, which had been unsuccessful by any other method.

Acetates↗

Toxic action of copper on the membrane system of a marine diatom measured by flow cytometry.

Flow cytometric measurements were used to investigate the toxic action of copper on some Phaeodactylum tricornutum membrane systems. Throughout the time of metal exposure, the percentage of viable cells decreased as copper concentration increased. The forward scatter signal increased as a result of copper exposure. After 72 h of metal exposure, cultures with 0.5 and 1 mg l-1 of copper showed an important increase in the peroxidase activity in comparison with control cells. Cells cultured with copper presented alterations in the membrane potential, increasing as copper concentration increased, after 96 h of metal exposure. Results obtained in this work showed that copper induced a degenerative process in P. tricornutum cells, closely related with alternations or disorders in membrane systems.

Cell Membrane↗

Flow cytometry determination of acute physiological changes in a marine diatom stressed by copper.

Flow cytometry (FCM) was used to determine changes in cellular volume, transmembrane potential, mitochondrial membrane potential and intracellular pH in the marine microalga Phaeodactylum tricornutum immediately (5 to 10 seconds) after the addition of selected concentrations of copper. An acute increase in the forward scatter signal of this diatom was detected after the addition of 10 mg 1-1 of copper. Stress produced by the copper addition resulted in various physiological alterations that can be easily and quickly detected by FCM: (i) the hyperpolarization of the cell membrane, as a result of an immediate increase in the cytoplasmic membrane potential, (ii) the increase of the mitochondrial membrane potential, being maximum at the higher copper concentration assayed, and (iii) the increase in the intracellular pH with the highest copper concentration assayed (10 mg 1-1).

Adaptation, Physiological↗

Different responses of the marine diatom Phaeodactylum tricornutum to copper toxicity.

Different responses of the marine diatom Phaeodactylum tricornutum (Bohlin) to toxic copper concentrations were investigated. Besides the classical variables applied to toxicity studies in microalgae, such as growth or chlorophyll a content, other variables analyzed by flow cytometry were used. Toxic effects due to copper concentration were observed. Cell density reached in the stationary phase was reduced to 50% in cultures with 20 mg Cu/l, with respect to control cultures without copper. Cell light scatter properties (related to cell volume and intracellular granularity) and chlorophyll a fluorescence of microalgal cells were determined by flow cytometry analysis at the beginning of growth, 1 h after copper exposure, and when cultures reached the stationary phase (72 h). After 1 h of exposure to metal, no differences were observed, but when cultures reached the stationary phase, a gradual increase in the variables analyzed by flow cytometry was observed as the copper concentration increased. The increase in chlorophyll a fluorescence detected by flow cytometry was not correlated with an increase in the cell content of this photopigment, thus indicating an inhibitory effect of copper on photosystem II.

Chlorophyll↗