Field applications of the second‐generation Environmental Sample Processor (ESP) for remote detection of harmful algae: 2006‐2007

Abstract

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.*Corresponding author: E-mail: dgreenfield@belle.baruch.sc.edu AcknowledgmentsWe thank the engineering technicians and machinists at MBARI for their invaluable help and dedication toward instrument development. We also greatly appreciate the hard work and dedication from the crew of the R/V Zephyr and Moss Landing Marine Laboratory, Small Boat Operations, and help from Drs. W. Jones and C. Preston. The authors gratefully acknowledge Drs. Z. Wang and T. Roth for the extraction and analysis of domoic acid in discrete water samples. This project was funded in part by the Monterey Bay Aquarium Research Institute from funds allocated by the David and Lucille Packard Foundation, the National Science Foundation, and the National Aeronautics and Space Administration (OCE-0314222, CCF424599, and NNG06GB34G to CAS, and OCE-0314089 to GJD).Disclaimers: Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the National Science Foundation. This publication does not constitute an endorsement of any commercial product or intend to be an opinion beyond scientific or other results obtained by the National Oceanic and Atmospheric Administration (NOAA). No reference shall be made to NOAA, or this publication furnished by NOAA, to any advertising or sales promotion which would indicate or imply that NOAA recommends or endorses any proprietary product mentioned herein, or which has as its purpose an interest to cause the advertised product to be used or purchased because of this publication. Limnol. Oceanogr.: Methods 6, 2008, 667-679 © 2008, by the American Society of Limnology and Oceanogra...

Metatranscriptomic analysis of autonomously collected and preserved marine bacterioplankton

“…The ESP is an automated fluid handling system that collects and processes biological samples from seawater (Scholin et al, 2009). Current real-time capabilities include array-based detection of target organisms including harmful algal species (Greenfield et al, 2006(Greenfield et al, , 2008Haywood et al, 2007), invertebrate larvae (Goffredi et al, 2006;Jones et al, 2008) and major bacterial and archaeal clades (Preston et al, 2009). Quantitative PCR capabilities are also currently in development (Scholin et al, 2009).…”

“…Quantitative PCR capabilities are also currently in development (Scholin et al, 2009). The ESP can return samples preserved in a saline-ethanol solution, but sample analysis has been primarily limited to in situ hybridization techniques (Goffredi et al, 2006;Greenfield et al, 2006Greenfield et al, , 2008Jones et al, 2008). To extend the scope of laboratory analysis of archived samples, new protocols for sample preservation were developed for gene expression analysis and transcriptomics.…”

Application of a sandwich hybridisation assay for rapid detection of the Northern Pacific seastar, Asterias amurensis

“…The SHA chemistry has been coupled with an in situ instrument, the Environmental Sample Processor (ESP), for autonomous environmental monitoring for a range of larval species simultaneously ). The SHA chemistry and the ESP have also been used to remotely detect marine bacterioplankton, dinoflagellates and invertebrates (Greenfield et al 2008;Preston et al 2009;Scholin et al 2009). …”

Molecular genetic tools for environmental monitoring of New Zealand's aquatic habitats, past, present and the future

“…The ESP also measures environmental parameters such as water temperature, salinity and turbidity. Laboratory and field trials have demonstrated near-real time detection of bacteria, harmful algal bloom species, the algal biotoxin domoic acid and invertebrates (Greenfield et al 2008;Jones et al 2008;Preston et al 2011;Ryan et al 2011;Harvey et al 2012). The MBARI ESPs have been trialled in many coastal waters including Monterey Bay, Florida, Massachusetts and Hawaii.…”

“…Traditionally, the ESP has employed SHA arrays that are very robust and allow the simultaneous detection of tens to hundreds of unique organisms in a single sample (Greenfield 2008;Harvey et al 2012). However, this method is not very sensitive and may miss the detection of harmful microbes at low densities.…”