Non-point Source Contaminant Monitoring and Consequent Genetic Response in Salmon

The Stillaguamish Tribe was concerned with chronic effects on Chinook salmon from low-level environmental exposure to emergent contaminants and with mixtures of these compounds that may have synergistic toxic effects. Specifically, the Tribe was concerned with the potential for intersex/sex reversal in Chinook as a result of wastewater effluent.

This project had two phases: Phase I was the completion of the water sampling effort by incorporating stormwater and agricultural runoff to the existing emergent contaminant chemistry databases. Phase II quantified genetic responses in juvenile salmon following exposure to emergent contaminants. Both phases were completed through a cost-share partnership with the U.S. Geological Survey (Patrick Moran, Richard Wagner and David Alvarez). Phase I involved the installation, extraction and analysis of Polar Organic Chemical Integrated Samplers (POCIS) and Phase II involved the caging of fish, organ extraction, and tissue analysis. The preferred approach was to place hatchery-raised juveniles (reared under specific conditions) in stationary cages at select sites in the watershed. This component of the sampling addressed the contaminant signature in caged juvenile salmonids as well as their gene response. Replicates of at least 3 fish from each location were used in profiling the gene response, and 2-3 of those same fish were analyzed for concentrations in their tissues. This focus was to link contaminant exposure to physiological pathways or ‘modes of action’ in juvenile fish.

While not all contaminants elicited a physiological response, evaluation of the gene response followed two paths, 1) what were the strongest physiological signals expressed in the gene response, and 2) were any of these gene responses predictable based on the contaminant profile in water or fish tissue. This involved interpretation and description of the gene response data in light of our current toxicological understanding.

Accomplishments:
• Completed a Draft workplan for field sampling in 2011, 2012, and 2013.
• Compiled existing water quality data for the Stillaguamish Watershed.
• Designed and constructed fish cages.
• Conducted a pilot project deploying caged juvenile Chinook salmon and Polar Organic Chemical Integrated Samplers (POCIS) at six study sites and one control site, which included sample extraction and analysis.

Fish cages and POCIS devices were deployed between April 11, 2011 and May 5, 2011.

Study Sites
In partnership with the USGS (Patrick Moran, Richard Wagner, and David Alvarez), the tribal lead biologist and a technician selected six sample sites and identified a control site at the Tribe’s Harvey Creek Hatchery (Figure 1). Sites were selected based on estimated land use by sub-basin for each stream of interest. The Tribe was interested in sampling streams that are influenced by agriculture, urbanization/industry, rural development and forestry.

The fish tissue portion of the project involved juvenile Chinook salmon raised at the Tribe’s Harvey Creek Hatchery. A sample of five of these study fish were taken at the start of the project to determine their baseline condition in terms of tissue and genetics. The Harvey Creek Hatchery was also used as the control site with fish cages and a passive sampler deployed in a trough adjacent to the trough that held the study fish. The methods for this part of the project were developed with assistance from the USGS, Western Washington University and the Northwest Indian Fisheries Commission’s Fish Pathologist (Craig Olsen).

This study is the next step in the Tribes effort to conduct a basin-wide assessment of the source and transport of contaminants that could have significant impacts on salmon health, productivity and survival. Project results should provide a list of emergent contaminants found in wastewater entering the Stillaguamish River. The Tribe will then work with USGS geneticists to quantify specific genetic responses in juvenile salmonids. The project will be the first of its kind to sample a membrane treatment system for contaminants in both influent and effluent. Data will indicate which compounds are being filtered or settled out of wastewater before it is released into the estuaries of Port Susan.

Water quality in the Stillaguamish Watershed impacts all living things. To date, watershed groups have been concerned only with temperature, specific conductance, total suspended solids, pH and fecal contamination. This project is part of a critical first step to identify other types of contaminants that pose a significant threat to humans and the environment. Once ECs are identified and sources are understood, we can begin to reduce the occurrence of these compounds and study their specific physiological impacts to species like salmon. Because the Tribe has an on-going MOU with the USGS and verbal agreement from the Warm Beach Wastewater Treatment Plant, this project is ready for implementation and has sampling methods and quality assurance/control procedures already in place. The USGS Water Science Center in Tacoma and Lakewood Colorado are currently working with the Tribe to analyze samples and compile data.

Proposed Outcomes/Benefits:
· Identify emergent contaminants in the waste-water influent and effluent;
· Provide baseline data for future upgrades that may occur at the plant;
· Identify if contaminants are being released into the Port Susan estuary;
· Build on a September 2008/09 effort to identify possible sources of emergent contaminants in the watershed;
· Generate data on contaminants that may impact juvenile salmon;
· Generate monitoring results that may be used to develop more thorough scientific studies of the types, sources, and fate of contaminants in the Stillaguamish watershed and other watersheds throughout the state; and
· Develop information useful for informing and educating water resources stakeholders, managers, scientists, and the public about emergent contaminant issues.