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

Alteration of Residual Circulation Due to Large-Scale Infrastructure in a Coastal Plain Estuary

Large-scale human-built infrastructure is shown to alter the salinity and subtidal residual flow in a realistic numerical simulation of hydrodynamic circulation in a coastal plain estuary (Tampa Bay). Two model scenarios are considered. The first uses a modern bathymetry and boundary conditions from the years 2001–2003. The second is identical to the first except that the bathymetry is based on depth soundings from the pre-construction year 1879. Differences between the models\u27 output can only result from changes in bay morphology, in particular built infrastructure such as bridges, causeways, and dredging of the shipping channel. Thirty-day means of model output are calculated to remove the dominant tidal signals and allow examination of the subtidal dynamics. Infrastructure is found to steepen the mean axial salinity gradient ∂s¯¯¯/dx by ~40% when there is low freshwater input but flatten ∂s¯¯¯/dx by ~25% under more typical conditions during moderate freshwater inflow to the estuary. Deepening of the shipping channel also increases the magnitude of the residual Eulerian circulation, allowing for larger up-estuary salt transport. Local bathymetry and morphology are important. Some regions within the estuary show little change in residual circulation due to infrastructure. In others, the residual circulation can vary by a factor of 4 or more. Major features of the circulation and changes due to infrastructure can be partially accounted for with linear theory

Phenological Changes of Blooming Diatoms Promoted by Compound Bottom-Up and Top-Down Controls

Understanding phytoplankton species-specific responses to multiple biotic and abiotic stressors is fundamental to assess phenological and structural shifts at the community level. Here, we present the case of Thalassiosira curviseriata, a winter-blooming diatom in the Bahía Blanca Estuary, Argentina, which displayed a noticeable decrease in the past decade along with conspicuous changes in phenology. We compiled interannual field data to assess compound effects of environmental variations and grazing by the invasive copepod Eurytemora americana. The two species displayed opposite trends over the period examined. The diatom decreased toward the last years, mainly during the winters, and remained relatively constant over the other seasons, while the copepod increased toward the last years, with an occurrence restricted to winter and early spring. A quantitative assessment by structural equation modeling unveiled that the observed long-term trend of T. curviseriata resulted from the synergistic effects of environmental changes driven by water temperature, salinity, and grazing. These results suggest that the shift in the abundance distribution of T. curviseriata toward higher annual ranges of temperature and salinity—as displayed by habitat association curves—constitutes a functional response to avoid seasonal overlapping with its predator in late winters. The observed changes in the timing and abundance of the blooming species resulted in conspicuous shifts in primary production pulses. Our results provide insights on mechanistic processes shaping the phenology and structure of phytoplankton blooms