Enumeration of cell mortality in the harmful bloom-forming dinoflagellates Karenia mikimotoi and Heterocapsa circularisquama using image cytometry
2016
Tomaru, Y. (National Research Institute of Fisheries and Environment of Inland Sea, Fisheries Research Agency, Hatsukaichi, Hiroshima (Japan)) | Kimura, K. | Yamaguchi, H.
We developed a new protocol for detecting and enumerating viable and dead cells of the potentially harmful, bloom-forming dinoflagellates, Karenia mikimotoi and Heterocapsa circularisquama. The method employs image cytometry and SYTOX-green, which binds specifically to the nucleotides of dead cells and can be used to simultaneously count the number of viable and dead cells of these dinoflagellates. In this study, we evaluated increases in cell mortality due to salinity stress and virus infection in K. mikimotoi and H. circularisquama, respectively. In K. mikimotoi, a marked increase in cell death was observed when the salinity was decreased rapidly from 30 to <18. Similarly, viral inoculation appeared to increase the number of dead H. circularisquama cells in experimental cultures. It is considered that cell mortality in these algae may have been induced by changes in a variety of physical, chemical and biological factors. Further studies using this protocol would facilitate a deeper understanding of the processes affecting disintegration of harmful algal blooms.
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