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

Recruitment and commercial seed procurement of the blue mussel Mytilus edulis in Maine

A thorough understanding of recruitment in the blue mussel is necessary if the new industry is to maximize seed procurement without impinging upon other fisheries. Larval appearance is a relatively precise event in Maine, cued to early summer water temperature of 10–12 C and, apparently, full moon spawning events. Mussel larvae are more abundant on the flood tides indicating inshore and estuarine retention, although this retention relates to the morphometry and relative energy of the system. Webb Cove, a wide embayment with maximum sample station current velocity of 0.2 m/s, showed a random ebb tide vs. flood tide larval distribution; the narrow, long Damariscotta River estuary with 0.35 m/s current velocity showed a two‐fold flood tide larval enhancement and the Jordan River with 1.5 m/s current velocities showed up to a 14‐fold flood tide enhancement of mussel larvae and bysally drifting juveniles. Thus certain Maine estuaries may act as larval traps, providing areas of concentrated settlement and seed abundance. Primary setting normally begins with a large initial pulse in June followed by one or more secondary pulses throughout the summer. Secondary settlement (reattachment of bysally drifting juveniles) occurs at lower levels throughout the year, especially in late July and early August. Maximum attachment of larvae and juveniles occurs during periods of maximum current velocity. Extensive eelgrass beds at the mouths of some estuaries (i.e., Jordan River) may be the sites of extensive primary and secondary setting. Great Eastern Mussel Farms, the industry component, guided by these studies, is testing the deployment of live and shell mussel cultch to develop and optimize a new seed procurement system