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The Columbia River has a dynamic and fast flushing estuary impacted by strong advection and mixing of riverine and oceanic waters, and high but variable loads of suspended particulate matter. Transient, but recurring water and nutrient fluxes from end-members impart strong spatial and temporal gradients, contributing to microbiological hotspots that play important ecological roles in the estuary. Investigations of corresponding microbiota require precisely timed samples that are contextualized by physical and biogeochemical data. To accomplish this, we embedded a robotic microbial sampler (Environmental Sample Processor, ESP) within the operations of an interdisciplinary observation and prediction system (Science and Technology University Research Network, SATURN; www.stccmop.org/saturn). Autonomous, adaptively sampled water collection by the ESP was implemented based on environmental conditions assessed from SATURN physical and biogeochemical sensors. Water was pumped from multiple depths to sensors and the ESP on dry land. If water met user-defined parameters, ESP sampling was automatically initiated. This strategy was tested during three deployments in summer 2013, during which operational tools for analysis and visualization were used to formulate well-constrained mission plans by providing estimates of the intensity and timing of oxygen-depleted ocean water intrusion and estuarine turbidity maxima. This allowed the effective characterization of the impact of these events on selected estuarine microbiota.

We assess the application of the second-generation Environmental Sample Processor (ESP) for the detection of harmful algal bloom (HAB) species in field and laboratory settings using two molecular probe techniques: a sandwich hybridization assay (SHA) and fluorescent in situ hybridization (FISH). During spring 2006, the first time this new instrument was deployed, the ESP successfully automated application of DNA probe arrays for various HAB species and other planktonic taxa, but non-specific background binding on the SHA probe array support made results interpretation problematic. Following 2006, the DNA array support membrane that we were using was replaced with a different membrane, and the SHA chemistry was adjusted. The sensitivity and dynamic range of these modifications were assessed using 96-well plate and ESP array SHA formats for several HAB species found commonly in Monterey Bay over a range of concentrations; responses were significantly correlated (p < 0.01). Modified arrays were deployed in 2007. Compared to 2006, probe arrays showed improved signal:noise, and remote detection of various HAB species was demonstrated. We confirmed that the ESP and affiliated assays can detect HAB populations at levels below those posing human health concerns, and results can be related to prevailing environmental conditions in near real-time.

A primary limitation of phytoplankton ecology research is the difficulty of describing patchiness and distributions of different phytoplankton groups. Chlorophyll fluorescence and optical backscatter are useful measurements that provide information about phytoplankton, but these measurements do not allow distinction of phytoplankton taxa. Traditional phytoplankton identification methods (such as microscopy, HPLC analysis, and flow cytometry) are labor intensive and therefore can provide only very limited coverage and resolution. Through lab experiments we show that the Laser In Situ Scattering and Transmissometer (LISST-100) instrument can accurately quantify phytoplankton cell dimensions for some cell shapes. Pseudo-spherical dinoflagellates are described with a single peak in the particle size distribution (PSD) at the cross-sectional dimension of the cells. Pennate diatoms are described with peaks in the PSD at the major and minor axis dimensions of the cells. Diatom cells with minor axis dimensions that vary along the major axis are described with one peak across the range of minor axis dimensions and a second peak at the major axis dimension. Through field experiments, we show that mapping the PSD in situ at high resolution permits description of patchiness and evolution of phytoplankton populations. We present two examples: (1) growing dominance of Ceratium species during a red tide bloom, and (2) concentration of Pseudo-nitzschia australis, a harmful algal bloom (HAB) species, at a water mass front. We conclude that synoptic mapping of the PSD can significantly advance phytoplankton ecology research in the coastal ocean.

In this contribution, we assess the application of methodology used in a novel in situ remote sampling platform, the environmental sample processor (ESP), for the identification and enumeration of a harmful algal species, the domoic acid-producing diatom Pseudo-nitzschia australis, using 2 molecular assays: a sandwich hybridization assay (SHA) and fluorescent in situ hybridization (FISH). Both the SHA and FISH assays were initially designed as laboratory-based methods that are now emulated in the ESP. Response of the SHA to a range of concentrations of P. australis using the laboratory (96-well plate) and ESP (DNA probe array) formats showed that the two were highly correlated. Enumeration of cells filtered and archived for FISH using a manifold designed for laboratory applications agreed well with counts of cells filtered and archived in the ESP at ≥2.5 × 104 cells L−1. At lower concentrations, it becomes statistically unlikely to derive a reliable abundance estimate, suggesting that FISH is better suited for qualitative analyses unless the target organism is very abundant. We also assessed the suitability of an oligonucleotide as synthetic target sequence to act as a SHA reagent quality control and internal standard for the plate assay. This was successful, but the probe and/or associated reagents were stable for only ~60 days. Our results show that the ESP is capable of detecting P. australis in near real-time and also supports whole-cell archival samples, making it a potentially useful tool for future research and monitoring initiatives.