GRANT
NUMBER: NA76FD0051
NMFS
NUMBER: 96-SWR-016
REPORT
TITLE:
Rapid Detection of Harmful Algal Bloom Species and Their
Associated Toxins using DNA Probes and a Receptor Binding
Assay
AUTHOR:
Christopher A. Scholin and Gregory J. Doucette
PUBLISH
DATE:
July 10, 2000
AVAILABLE
FROM:
National Marine Fisheries Service, Southwest Region, 501
West Ocean Blvd., Suite 4200, Long Beach, CA 90802-4213.
TELEPHONE: (562) 980-4033
ABSTRACT

This project
focused on the challenge of obtaining nearreal-time
data on the abundance (spatial and temporal) and toxicity
of algal cells for the routine monitoring and scientific
investigation of harmful algal blooms and their associated
toxins. The primary impediment to obtaining such information
is a lack of appropriate, field-tested methods. This project
was designed to resolve this problem by evaluating the
performance of two techniques that lend themselves to
rapid detection of algal cells and their toxins: species-specific
DNA probes and phycotoxin receptor binding assays, respectively.
The project focused on populations of domoic acid (DA)-producing
diatoms of the genus Pseudo-nitzschia that occur
in Monterey Bay, California. Species-specific probes targeting
Pseudo-nitzschia were applied using both whole-cell
and sandwich hybridization methods, with good agreement
between methods observed during the early stages of bloom
development when cells were actively growing. Although
some discrepancies arose toward the latter phase of a
bloom, due largely to underestimates using the whole-cell
technique with cells in poor condition, both approaches
were generally effective in detecting toxic cells at concentrations
relevant for routine monitoring. The receptor binding
assay reliably detected DA associated with cells during
periods of bloom development, but the assay occasionally
revealed DA in the absence of recognizable, intact cells
as blooms declined. Precise estimates of cell abundance
and toxicity may not be required for routine monitoring.
Perhaps most important for public health officials and
wildlife biologists are up-to-date trends of where potentially
toxic Pseudo-nitzshia species are; whether the
populations are on the rise, falling, or exceeding some
thresold value; and whether any associated toxicity warrants
attention. In this regard, sandwich hybridization application
of DNA probes for Pseudo-nitzschia and the DA receptor
binding assay are technologies that show good promise
and should be tested further against currently used methods
in collaborative trials.