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Domoic Acid Test Kit Helps Tribes in the Pacific Northwest


Domoic acid test kit workshop participants from the Quinault Nation, Quileute Tribe and University of Washington Olympic Natural Resources Center, Northwest Fisheries Science Center, Oregon Department of Fish and Wildlife, Center for Coastal Fisheries and Habitat Research (Pat Tester, NOAA)

Better. Faster. Easier.

And cheaper too.

It might sound like the latest and greatest gizmo being hawked over late–night TV.

But those are actual terms that end–users—read that as "customers"—apply to their experiences with a new testing method developed by National Centers for Coastal Ocean Science researchers for detecting and measuring domoic acid.

The new domoic acid test kits are saving Pacific Northwest coastal communities and fishers effort and, most importantly, valuable time in assuring the safety and vitality of a coastal resource they depend on, both for food and for tourism income.


Technicians from the Californian Department of Health Services running the domoic acid ELISA. (Pat Tester, NOAA)

"We could not be more enthusiastic to see this technology go forward and become readily available to the coastal tribes and other communities," says Joe Schumacker, marine resource scientist with the Quinault Indian Nation in the Pacific Northwest.

"With our geography, we’re really rural and remote out here, about two and a half hours away from Seattle, more like three hours. And that’s where all the testing and regulatory agencies are located," Schumacker said in a phone interview.

Not so long ago, all Quinault samples used in detecting and measuring domoic acid—a harmful marine algal toxin that can cause serious illness and death in humans and marine mammals—had to be shipped off to Seattle for testing. For regulatory approval pending final validation and for those kits being used for that validation, that is still the rule.


Jonnette Bastian, Harmful Algal Bloom Specialist with the Quinault Indian Nation, in her environmental laboratory at Taholah, WA (Pat Tester, NOAA)

Even when the round–trip transportation and the regulatory agencies’ testing schedules works perfectly, "we needed to send the shellfish or seawater samples to Seattle overnight, have them tested there on their schedule, and then sent back on their schedule. At its best, it’s 48 hours," Schumacker said, unless they end up delivering them first–hand themselves, an increasingly costly option given the historic rise in the price of gasoline.

"Now we have a timely way to determine whether there is an issue with our clams out here," and validation studies are under way to assure compliance with Washington State Department of Health standards and eventual Food and Drug Administration (FDA) approval. The newly developed ELISA test kits (Enzyme–Linked ImmunoSorbent Assay) now allows them to process samples and analyze results in the convenience of their own on–site laboratories…and have results back within one and a half or two hours of the harvest.


The topic of the day was the domoic acid ELISA training and reminding everyone that clam hunger has its own word in the Quinault language. (Pat Tester, NOAA)

That reference to "our clams" is an important one. Razor clams occupy an especially important role in the Quinault culture, so much so that there is a specific word meaning "clam hungry" (ta’a Wshi xa’iits’os). Closures of coastal shellfish beds and shellfishing waters to protect public health can impair an important subsistence resource for coastal tribes such as the Quinault and the Quileute. And those closures, whether in fact they actually turned out to have been justified based on the testing results, have led to commercial losses for coastal communities dependent on their $12 million annual shellfishing and tourism economy.

Asked about the economics of using the new domoic acid test kits and testing protocol, Schumacker pointed to up–front costs of about $20,000 for a plate reader, plate washer, and pipettes; and another $5,000 for a centrifuge and supplies.


Jonnette Bastian shucking razor clams for the domoic acid test kit demonstration at Taholah, WA (Quinault Department of Fisheries)

The University of Washington ORHAB group ended up purchasing the plate reader and washer for the study and placing them in the Quinault laboratory. "We expect over time to recover all our expenses and save both time and money on shipments to Seattle," Schumacker said. "Our costs will work out fine."

Domoic acid is one of the two most important biological toxins posing risks to west coast razor clams and other bivalves. The product of a group of naturally occurring diatoms which occasionally bloom off the Pacific coast, domoic acid causes amnesiac shellfish poisoning (ASP). Persons ingesting tainted razor clams, oysters, crab, and mussels generally demonstrate symptoms—vomiting, nausea, diarrhea, abdominal cramps, headaches, confusion, dizziness, and short–term memory loss—after one to two days.


Homogenized razor clams prepared for extraction and domoic acid analysis. (Quinault Department of Fisheries)

Under the testing protocol and technology developed by an unusual partnership including federal agencies, tribal interests, and a small biotech company, the ELISA methodology allows the Quileute Tribe and Quinault Nation and other native American tribes across the region to rapidly detect toxin levels, evaluating razor clam samples for toxins in less than one and a half hours and providing real–time data on toxin levels and potential risks. Now being proven to be accurate, easy to use, quick, and reliable—while at the same time being less expensive—the advance is reducing per–sample expenses from around $150 to about $10.


Extraction of domoic acid from razor clam homogenate. (Quinault Department of Fisheries)

Behind the innovative research and manufacturing/distribution collaboration lies a far–sighted and somewhat unusual partnership spanning the tribal interests, diverse federal and state agencies and their often independent line offices, and a private sector laboratory partner. An especially interesting aspect of the partnership allows for some sales proceeds to be returned directly to the NCCOS laboratory that led the project—the Center for Coastal Fisheries and Habitat Research, in Beaufort, N.C.

"The project effectively paired the needs of the Pacific Northwest coastal communities and our private sector laboratory’s ability to meet those specialized needs in a cost–effective way," said Mercury Science, Inc., President Tom Stewart, commenting on the test kit partnership.


Centrifuge for separating particles of razor clams from the liquid extraction phase. (Quinault Department of Fisheries)

Speaking specifically of the leadership roles of the most actively involved NCCOS scientists, Stewart said "the goals were clear; there were effective personal–level good communications throughout; and all parties kept their ends of the bargain in moving forward. Each step in the scientific process led clearly to the next step, and we never came to the point of wondering what happens next."

Playing key roles throughout the entire development effort were NCCOS scientists from the Beaufort facility; scientific and management experts from NCCOS’s Monitoring and Event Response for Harmful Algal Blooms (MERHAB)-sponsored research program, scientific experts from NOAA, the National Ocean Service, and the National Marine Fisheries Service, representatives from the Quileute Tribe and the Quinault Nation; expert legal counsel from the U.S. Department of Commerce; officials from California’s State Department of Health and from the University of California, Santa Cruz; officials from the Washington State Department of Health, Office of Shellfish and Water Protection, from Oregon state agencies, and from Mercury Science, Inc.


Pipetting razor clam extract for testing using the domoic acid test kit. (Quinault Department of Fisheries)

Sharing Stewart’s and Schumacker’s enthusiasm for both the process they followed and also the resulting Domoic Acid ELISA test kit protocol, NCCOS scientist Patricia Tester said "the right combination of things had to happen. The effort could have been stopped at any number of places along the way. There were a lot of critical points."

But in the end, Tester said, "The story was so compelling that everybody wanted to entrain into this activity. This is what we train for, this is what we want to do."

In recognition of their effort, NOAA in late 2007 awarded NCCOS scientists Wayne Litaker, Pat Tester, and Marc Suddleson the Technology Transfer Award, specifically acknowledging them for "development and commercialization of rapid, cost–effective detection of algal toxins threatening human health and marine resources in coastal waters." Along with the three NCCOS scientists, the award also was given to NMFS scientists Vera Trainer, Bich–Thuy Eberhart, and John Wekell (retired).


96-Well plate reader provides precise measurements of color changes todetermine concentrations of domoic acid. (Quinault Department of Fisheries)

"We really could not have done this without the generous donation of the kits from the Beaufort Laboratory and the help of the Mercury Science folks," Quinault marine resource scientist Schumacker said in summing up the widespread collaboration. "The Quinault Nation did not have the funds necessary to purchase the test kits we used in the validation study, and Pat Tester and Wayne Litaker, in particular, were wonderful in having them supplied to us. Their willingness to work with us as end–users was key to this successful effort: We needed better testing ability, and they needed a good place to conduct field trials. Win–win!"


Domoic acid analysis results from plate reader. (Quinault Department of Fisheries)

"It really was a perfect storm," Stewart agreed. "A lot of people were in the right place at the right time."

Add to that, he said, "It’s always more fun to do science that’s helpful to people, as was the case here."

Citation

Litaker, R.W., T.N. Stewart, B.–T.L.Eberhart, J.C. Wekell, V.L. Trainer, R.M. Kudela, P.E. Miller, A. Roberts, C. Hertz, T.A.Johnson, G. Frankfurter, G.J. Smith, A. Schnetzer, J. Schumacker, J. L. Bastian, A. Odell, P. Gentien, D. LeGal, D. R. Hardison and P.A. Tester. 2008. Rapid enzyme–inked immunosorbent assay for detection of the algal toxin domoic acid. Journal of Shellfish Research 27(5): 1–10.