The marine microalga Isochrysis galbana was cultured in nitrate, nitrite or urea media to examine changes in the gross biochemical composition, with special emphasis on the growth phase associated changes in the compositions of the lipid classes and fatty acids. The gross biochemical composition was affected more by the growth phase than by the nitrogen source.Protein was higher during exponential growth (about 37-45% AFDW) but the pattern changed as the cultures aged, and lipids were the main algal constituents on all nitrogen sources in the late stationary phase. In all cultures, the relative abundance of neutral lipid increased in the late stationary phase concomitant with a proportional reduction in phospholipids, whereas galactolipids only slightly changed during the growth of the cultures. Total fatty acid content was affected by nitrogen source and growth phase. Maximal PUFA values were obtained at the early stationary phase and decreased throughout the stationary phase. The highest fatty acid contents in the early stationary phase were produced in urea cultures; these cultures also had higher PUFA content, with EPA and DHA contents of 27.66% and 14.13% of total fatty acids, respectively.
Essential heavy metals, as copper, can be toxic for microalgae at high concentrations. Copper affected growth and other parameters closely related to photosynthesis of the marine diatom Phaeodactylum tricornutum. A copper concentration of 0.10 mg l −1 provoked about 50% growth reduction and 1 mg l −1 inhibited the growth. Copper also interfered with photosynthesis and ATP production. A copper concentration of 0.5 mg l −1 reduced in a 50% the photosynthetic rate. Therefore, growth is more affected by copper than photosynthesis. Results of chlorophyll a fluorescence obtained by flow cytometry showed that copper's inhibitory effect on PS II activity is located on its oxidizing side. The lower copper concentration assayed provoked a significant decrease in the cellular pool of ATP. Pigment analysis by HPLC showed that copper affected the pigment pattern of P. tricornutum. Important changes were observed for chlorophyll a and its allomer: chlorophyll a proportion decreased while its allomer increased with the copper concentration, being maximum at 1 mg Cu l −1. The study of the intracellular pH by flow cytometry revealed that P. tricornutumcells exposed to 0.5 and 1 mg Cu l −1 showed an intracellular pH higher than control cultures cells, explaining the high proportion of the chlorophyll a allomer in these cells.
Cadmium removal by living cells of the marine microalga Tetraselmis suecica was tested in cultures exposed to different cadmium concentrations (0.6, 3, 6, 15, 30 and 45 mg/l). The EC50 for growth was 7.9 mg Cd/l after six days of exposure. The cadmium removed was proportional to the concentration of this metal in the medium and it was dependent on the time of exposure; cultures with higher cadmium concentration removed a higher amount of this metal. In cultures exposed to 0.6 mg/l, T suecica cells removed 98.1% of added cadmium with 0.392 x 10(-6) microg Cd/cell, whereas in cultures with 45 mg/l only 7.7% was removed with 16.052 x 10(-6) microg Cd/cell. The highest amount of cadmium removed per liter of culture was observed in cultures exposed to 6 mg/l, with 3.577 mg/l of cadmium. After six days of incubation, the higher proportion of cadmium was bioaccumulated intracellularly in all cultures except in 45 mg/l cultures, the percentage of intracellular cadmium being always more than 50%. The highest percentage of bioadsorbed cadmium (60.1%) was found in cells of cultures with the highest cadmium concentration (45 mg/l). Furthermore, a relation between intracellular cadmium and the concentration of sulfhydryl groups was observed.
Certain marine microalgae contain water-and lipid-soluble vitamins and can be used as food supplements or food ingredients. A number of vitamins are present in higher concentrations in the microalgae than in conventional foods traditionally considered rich in them. Ingestion of relatively small quantities of microalgae can cover the requirements for some vitamins in animal nutrition, including human nutrition, while supplementing others. Marine microalgae can thus be considered to represent a non-conventional source of vitamins or a vitamin supplement for animal or human nutrition.
The use of herbicides constitutes the principal method of weed control but the introduction of these compounds into the aquatic environment can provoke severe consequences for non-target organisms such as microalgae. Toxic effects of these pollutants on microalgae are generally evaluated using phytotoxicity tests based on growth inhibition, a population-based parameter. However, physiological cellular endpoints could allow early detection of cell stress and elucidate underlying toxicity mechanisms. Effects of the herbicide paraquat on the freshwater microalga Chlamydomonas moewusii were studied to evaluate growth rate and cellular parameters such as cellular viability and metabolic activity assayed by flow cytometry and DNA damage assayed by the comet assay. Sensitivity of growth and parameters assayed by flow cytometry were similar, showing a significant effect in cultures exposed to a paraquat concentration of 0.1 microM or higher, although in cultures exposed during 48 h to 0.05 microM, a significant stimulation of cellular fluorescein fluorescence was observed, related to cellular metabolic activity. After only 24 h of herbicide exposure significant DNA damage was observed in microalgal cells exposed to all paraquat concentrations assayed, with a 23.67% of comets in cultures exposed to 0.05 microM, revealing the genotoxicity of this herbicide. Taking into account the results obtained, comet assay provides a sensitive and rapid system for measuring primary DNA damage in Chlamydomonas moewusii, which could be an important aspect of environmental genotoxicity monitoring in surface waters.
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