Comparison of techniques used to count single-celled viable phytoplankton

Author Posting. © The Author(s), 2010. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Journal of Applied Phycology 24 (2012): 751-758, doi:10.1007/s10811-011-9694-z.Four methods commonly used to count phytoplankton were evaluated based upon the precision of concentration estimates: Sedgewick Rafter and membrane filter direct counts, flow cytometry, and flow-based imaging cytometry (FlowCAM). Counting methods were all able to estimate the cell concentrations, categorize cells into size classes, and determine cell viability using fluorescent probes. These criteria are essential to determine whether discharged ballast water complies with international standards that limit the concentration of viable planktonic organisms based on size class. Samples containing unknown concentrations of live and UV-inactivated phytoflagellates (Tetraselmis impellucida) were formulated to have low concentrations (<100 ml-1) of viable phytoplankton. All count methods used chlorophyll a fluorescence to detect cells and SYTOX fluorescence to detect non-viable cells. With the exception of one sample, the methods generated live and non-viable cell counts that were significantly different from each other, although estimates were generally within 100% of the ensemble mean of all subsamples from all methods. Overall, percent coefficient of variation (CV) among sample replicates was lowest in membrane filtration sample replicates, and CVs for all four counting methods were usually lower than 30% (although instances of ~60% were observed). Since all four methods were generally appropriate for monitoring discharged ballast water, ancillary considerations (e.g., ease of analysis, sample processing rate, sample size, etc.) become critical factors for choosing the optimal phytoplankton counting method.This study was supported by the U.S. Coast Guard Research and Development Center under contract HSCG32-07- X-R00018. Partial research support to DMA and DMK was provided through NSF International Contract 03/06/394, and Environmental Protection Agency Grant RD-83382801-0

Detecting, Characterizing and Determining the Biological Response to Regime Shifts off the California Coast

First, using one method of change detection analysis called the cumulative sum, it is possible to detect and characterize regime shifts along the California coast using sea surface temperatures (SSTs) and other variables. Second, physically-determined regime shifts and changes in ocean climatology, determined largely through detailed temperature time-series, can be linked to corresponding changes in biological communities, particularly phytoplankton, which exhibit rapid generation times

Ecological dynamics in the subarctic Pacific, a possibly iron-limited ecosystem

It has been suggested that production in offshore waters of the subarctic Pacific is limited by availability of dissolved Fe. Although that is not yet adequately established, the functional consequences of the limitation (if it exists) can be characterized from the results of the Subarctic Pacific Ecosystem Research (SUPER) program. Fe limitation, or something like it, establishes a phytoplankton community dominated by very small cells. These plants are not limited by Fe availability. Rather, their production is limited by their stock and available illumination. Stock is set by microzooplankton grazers with rapid population growth rates and, thus, rapid response to increases in phytoplankton abundance. Micrograzers provide efficient recycling of nitrogen as NH4, and the ready availability of NH4 sharply limits the annual utilization of NO3. Persistently high NO3 concentrations result. Other possibly Fe-limited, oceanic ecosystems with persistently high, near surface nutrients require similar, detailed analysis of ecosystem function