|Olson, Brady M.||Western Washington University (WWU)||Principal Investigator|
|Love, Brooke||Western Washington University (WWU)||Co-Principal Investigator|
|Strom, Suzanne||Western Washington University (WWU)||Co-Principal Investigator|
|Copley, Nancy||Woods Hole Oceanographic Institution (WHOI BCO-DMO)||BCO-DMO Data Manager|
Kendall, K., Marine Microzooplankton are Indirectly Affected by Ocean Acidification Through Direct Effects on Their Phytoplankton Prey. (Masters Thesis) Western Washington University.
The phytoplankton Emiliania huxleyi CCMP 2668 was grown semi-continuously in atmosphere controlled chambers at three different CO2 treatment concentrations; Ambient (400ppmv), Moderate (750ppmv), and High (1000ppmv). Cultures were diluted daily starting day 4 with pre-equilibrated media containing f/50 nutrients. Each morning of dilution the cultures were gently mixed prior to a small sample being taken for cell counts. Cells were counted live on a BD FACSCalibur flow cytometer and were quantified based on side scatter and red fluorescence parameters. Prior to counting, a measured volume of 2µm yellow-green latex bead solution (Flow Check intermediate intensity level 1 fluorescence in nanopure water) was added to each sample of E. huxleyi. The bead solution was counted manually with epi-fluorescence microscopy using UV light. Cell concentration was determined by the ratio between cells and beads counted. The dilution volume was then calculated to achieve a cell density that would remain close to or under 100,000 cells per ml during the next day of growth.
The known bead solution standard was used to determine the actual volume that was analyzed in the flow cytometer for each sample. The events per ml was then calculated.
- added conventional header with dataset name, PI name, version date
- column names reformatted to comply with BCO-DMO standards
- transformed table columns to rows
|treatment||pCO2 level (ambient; moderate; high) and replicate (A; B; C)||unitless|
|culture_day||days the cells have been in culture: count day for the morning cell count or count day post-dilution for the predicted cell count after dilution||days|
|Dataset-specific Instrument Name|
|Generic Instrument Name|| |
|Dataset-specific Description|| |
BD FACSCalibur flow cytometer
|Generic Instrument Description|| |
Flow cytometers (FC or FCM) are automated instruments that quantitate properties of single cells, one cell at a time. They can measure cell size, cell granularity, the amounts of cell components such as total DNA, newly synthesized DNA, gene expression as the amount messenger RNA for a particular gene, amounts of specific surface receptors, amounts of intracellular proteins, or transient signalling events in living cells. (from: http://www.bio.umass.edu/micro/immunology/facs542/facswhat.htm)
|Start Date|| |
|End Date|| |
Description from NSF award abstract:
The calcifying Haptophyte Emiliania huxleyi appears to be acutely sensitive to the rising concentration of ocean pCO2. Documented responses by E. huxleyi to elevated pCO2 include modifications to their calcification rate and cell size, malformation of coccoliths, elevated growth rates, increased organic carbon production, lowering of PIC:POC ratios, and elevated production of the active climate gas DMS. Changes in these parameters are mechanisms known to elicit alterations in grazing behavior by microzooplankton, the oceans dominant grazer functional group. The investigators hypothesize that modifications to the physiology and biochemistry of calcifying and non-calcifying Haptophyte Emiliania huxleyi in response to elevated pCO2 will precipitate alterations in microzooplankton grazing dynamics. To test this hypothesis, they will conduct controlled laboratory experiments where several strains of E. huxleyi are grown at several CO2 concentrations. After careful characterization of the biochemical and physiological responses of the E. huxleyi strains to elevated pCO2, they will provide these strains as food to several ecologically-important microzooplankton and document grazing dynamics. E. huxleyi is an ideal organism for the study of phytoplankton and microzooplankton responses to rising anthropogenic CO2, the effects of which in the marine environment are called ocean acidification; E. huxleyi is biogeochemically important, is well studied, numerous strains are in culture that exhibit variation in the parameters described above, and they are readily fed upon by ecologically important microzooplankton.
The implications of changes in microzooplankton grazing for carbon cycling, specifically CaCO3 export, DMS production, nutrient regeneration in surface waters, and carbon transfer between trophic levels are profound, as this grazing, to a large degree, regulates all these processes. E. huxleyi is a model prey organism because it is one of the most biogeochemically influential global phytoplankton. It forms massive seasonal blooms, contributes significantly to marine inorganic and organic carbon cycles, is a large producer of the climatically active gas DMS, and is a source of organic matter for trophic levels both above and below itself. The planned controlled study will increase our knowledge of the mechanisms that drive patterns of change between trophic levels, thus providing a wider array of tools necessary to understand the complex nature of ocean acidification field studies, where competing variables can confound precise interpretation.