| Impact of Crude Oil and Dispersant on Determination of Calcium Concentration in Whole Cell from Cochlodinium polykrikoides
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| Eun Seob Cho, Dong Woo Kang, Yong Chul Cho
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| ABSTRACT |
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The effects of Kuwait crude oil and World clean 90 dispersant on Ca supper(++) flux in Cochlodinium polykrikoides were studied using flow cytometry and confocal microscope. Sugar composition and moieties on the cell surface were examined using fluorescent-tagged lectins. Cells showed no difference in forward light scatter and side-angle scatter (FSC-SSC) plot between control and crude oil or dispersant treated cells. In FSC and SSC, treatment with calcium ionophore (A23187) or without phorbol 12-myristate 13-acetate (PMA) or ionomycin showed that most of the plankton were dead and planktons treated with crude oil or dispersant also showed some dead cells, indicating a too high Ca supper(++) flux. Relative fluorescence intensity (RFI) in calcium labelling intensity showed that cells treated with dispersant had increased significantly in number, compared with crude oil. In lectin binding test, cell treasted with crude oil and dispersant gave the same positive response, even with exposure to a high concentration of crude oil and dispersant. Considering these results, treatment with high concentration of the dispersant cause a significant increase in Ca supper(++) influx, which effects the regulation of the transport ion channels across a membrane. Thus, the toxic effects of dispersant on C. polykrikoides at high concentrations were possibly because of an increase of calcium ion in the cells. Crude oil and dispersant did not effect sugar distribution on the cell surface.
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