Intertidal Forage Fish Spawning Surveys In

Quality Assurance Project Plan Intertidal Forage Fish Spawning Surveys in Three Washington State Aquatic Reserves

Washington Department of Natural Resources Ensuring regulatory effectiveness in Puget Sound’s most special places Grant #: PC-00J29801-0

Prepared for:

Fidalgo Bay, Nisqually Reach, and Smith and Minor Islands Aquatic Reserve Citizen Stewardship Committees

Prepared by:

Michael Grilliot Nisqually Reach Nature Center and Jerry Joyce Washington Environmental Council

June 2013 Publication Information

This project has been funded wholly or in part by National Estuary Program (NEP) of the United States Environmental Protection Agency (EPA) under assistance agreement PC-00J29801-0 to Washington Department of Natural Resources (WDNR). The EPA and Washington State Department of Ecology (Ecology) require a Quality Assurance Project Plan (QAPP) for studies, such as this one, that generate environmental data. This QAPP describes monitoring of environmental conditions that will be conducted in 2013 at three of Washington’s aquatic reserves. However, the contents of the QAPP do not necessarily reflect the views and policies of the Environmental Protection Agency, nor does mention of trade names or commercial products constitute endorsement or recommendation for use. Copies of this QAPP and final project publications will be available from the Washington State Department of Natural Resources (http://www.dnr.wa.gov/ResearchScience/Topics/AquaticHabitats/Pages/aqr_rsve_aqua tic_reserves_program.aspx).

Author and Contact Information Michael Grilliot 2525 Minor Ave E. Apt 410 Seattle, WA 98102 740-503-8111 [email protected] Jerry Joyce 11740 Exeter Ave NE Seattle, WA 98125 206-440-8688 [email protected]

2 Intertidal Forage Fish Spawning Surveys in Three Washington State Aquatic Reserves Quality Assurance Project Plan

June 2013

Approved by:

Signature: Date: Yvonne Shavalier, Principal Investigator (PI) Nisqually Reach Aquatic Reserve

Signature: Date: Daniel Hull, NRAR Citizen Stewardship Committee Signature: Date: Robin Clark, PI and Stewardship Committee, Smith and Minor Islands Aquatic Reserve Signature: Date: Pete Haase, PI and Stewardship Committee, Fidalgo Bay Aquatic Reserve Signature: Date: Wendy Steffensen, Staff Scientist, RE Sources Signature: Date: Kerri Cechovic, Washington Environmental Council Signature: Date: Jerry Joyce, Washington Environmental Council Signature: Date: Kyle Murphy, WDNR Signature: Date: Dion Jamieson, WDNR Signature: Date: William Kammin, QA Officer, Ecology

3 | P a g e Distribution List

Kyle Murphy, Program Manager Daniel Hull, PI WDNR Aquatic Reserves NRNC 1111 Washington Ave SE 4949 D’Milluhr Dr NE Olympia, WA 98504 Olympia, WA 98516 [email protected] 360- 459-0387 [email protected] Dion Jamieson WDNR Aquatic Reserves Kerri Cechovic, Grant Manager 1111 Washington Ave SE WEC Olympia, WA 98504 1402 Third Ave, Seattle, WA 98101 Pete Haase 206-631-2607 Beach Watchers [email protected] Fidalgo Bay Aquatic Reserve (FBAR) Citizen Stewardship Committee Jerry Joyce, Science Advisor [email protected] WEC 11740 Exeter Ave NE Yvonne Shevalier Seattle, WA 98125 NRAR 206-440-8688 Citizen Stewardship Committee [email protected] Volunteer lead (Mainland committee) [email protected] Tom Gries, NEP QA Coordinator Robin Clark Ecology Whidbey Watershed Stewards 300 Desmond Drive SE [email protected] Lacey, WA 98503 [email protected] Michael Grilliot, Board Nisqually Reach Nature Center (NRNC)

2525 Minor Ave E. Apt 410 Seattle, WA 98102 740-503-8111 [email protected]

4 | P a g e Table of Contents Page

Table of Figures

able of Tables

5 | P a g e Abstract As steward of 2.6 million acres of state-owned aquatic lands, the Washington Department of Natural Resources (WDNR) established aquatic reserves throughout Puget Sound to protect important native ecosystems. These reserves are an effort to promote the preservation, restoration, and enhancement of state-owned aquatic lands that are of special educational, scientific, or environmental interest. This project establishes forage fish beach surveys in three aquatic reserves in Puget Sound: Fidalgo Bay, Nisqually Reach, and Smith and Minor Islands. The sampling will be conducted by trained citizen-science volunteers that have been organized by each aquatic reserve citizen stewardship committee. Forage fish are a vital component in the Puget Sound ecosystem, and the monitoring of their status is an important component to the recovery of Puget Sound and the Salish Sea. The citizen scientist groups will use established Washington Department of Fish and Wildlife (WDFW) sampling and processing protocols. Surveys will be conducted monthly June- November and twice monthly beginning in December. If the program continues beyond the scope of this grant, sampling will be done twice monthly in January and February and once monthly from March through November. Retrieved samples will be returned to WDNR for identification and quantification of the forage fish eggs recovered in the study. Background The WDNR is steward of more than 2.6 million acres of aquatic lands. As steward, it has established aquatic reserves throughout Puget Sound to protect important native ecosystems. The Aquatic Reserves (AR) Program focuses on conserving high-quality native ecosystems. It is a state-wide effort to promote the preservation, restoration, and enhancement of state-owned aquatic lands that are of special educational, scientific, or environmental interest. One benefit of the AR Program is the partnerships WDNR establishes to aid in development and implementation of site-specific aquatic reserve management plans. WDNR works with federal, state, local, tribal and non-governmental organizations and private citizens in an effort to identify and manage important resources for conservation at each reserve. An additional benefit of AR designations is that management plans can be designed to compliment other protective measures within or adjacent to the reserve. The types of Aquatic Reserves include:  Environmental— promote conservation and restoration.  Scientific—provide unique aquatic habitats for research.  Educational—promote opportunities for field-based environmental education. Some of the anticipated benefits of these aquatic reserves include:

6 | P a g e  Ensuring environmental protection through site-based preservation, restoration, and enhancement.  Enhance the health of native marine and freshwater aquatic habitats, and the fish and wildlife that depend on them.  Encouraging public use and access.  Providing for greater public input into conservation management.  Working with stakeholders, including citizens and state, local and federal governments, to develop and implement site-specific management plans. WDNR plans to coordinate with citizen science groups to conduct spawning surveys in the Fidalgo Bay Aquatic Reserve (FBAR), Nisqually Reach Aquatic Reserve (NRAR), and Smith and Minor Islands Aquatic Reserves (SMAR). The Citizen Stewardship Committees (CSC) selected to conduct the research in these reserves due to importance of the habit for forage fish spawning while avoiding duplication of monitoring being conducted in other areas. The locations of these reserves are shown in Figure 1. Specific management plans were written for each of these reserves. These plans include requirements and recommendations for research and monitoring within these areas. The recommendations include: A. Identify data gaps, baseline inventory to fill gaps and establish standards for trend monitoring. B. Establish baseline conditions. C. Trend monitoring to determine the effectiveness of management activities and document natural variation. D. Research, to better understand observed changes and the interactions between management activities and natural resource conditions. Descriptions of each AR and their management plans are available from WDNR at the AR Program internet site http://www.dnr.wa.gov/ResearchScience/Topics/AquaticHabitats/Pages/ aqr_rsve_aquatic_reserves_program.aspx (accessed on January 17, 2013). In the development of the 2012-13 Action Agenda, the Puget Sound Partnership (PSP) established a near-term action for forage fish in Marine Protected Areas, including the aquatic reserves (PSP, 2012) B3.1 NTA 1: Marine Protected Area Effectiveness. By June 2014, PSP, in collaboration with WDFW and DNR will identify the threats, coverage gaps, and conservation concerns addressed by existing Puget Sound marine protected areas and assess the potential effectiveness of these MPAs to protect threatened species and habitats, including rockfish and forage fish. The importance of forage fish in the Puget Sound ecosystems has long been recognized. WDFW (1998) established a policy (POL-C3012) on the

7 | P a g e management of forage fish that requires management policies based on monitoring data. A poster by Pierce, et al (2012) summarizes much of the past sampling of forage fish in Puget Sound. A symposium on forage fish was also was also held in September 2012 (Liedtke, 2013) that both summarized current work and presented new research. One of the conclusions of the symposium was that “The lack of a reliable, and stock-specific, abundance estimate is a large data gap for most forage fish species in the Salish Sea. It is difficult to assess whether forage fish populations are stable, growing, or declining without a baseline population estimate and a means of assessing abundance on a relevant time scale. Herring are the only forage fish species in Puget Sound that are regularly monitored (by WDFW), and the group recommended that future effort be more balanced across all forage fish species.” Forage fish beach spawning surveys have been recognized as a reliable way of monitoring forage fish spawning abundance and determining critical spawning habitat sites. ( Moulton and Pentilla, 2001). Forage fish are also considered a major issue in the establishment of the aquatic reserves. In the Fidalgo Bay AR Management Plan (WDNR, 2008), it recognizes that a primary goal of establishing the reserve was to protect herring spawning habitat. Project Description The goals of the intertidal forage fish spawning surveys in the ARs are to:  Collect time sensitive baseline data throughout each reserve  Document changes over time in forage fish usage of the beaches using established methods that will provide data comparable across reserves and monitoring years. The objectives of this project are to:  Train and empower citizen scientists in conducting surveys using a standardized protocol  Implement the surveys on a regular and consistent basis that can contribute to the time series from previous monitoring studies conducted in these reserves and concurrent monitoring studies in other ARs and other regions of Puget Sound, as described in this QAPP.  Create a sustainable, locally operated structure in each reserve to continue this work after the end of this grant. Monitoring is essential to determine if and how forage fish are using beaches in the reserves. The natural and/or human-induced impacts on reserve beaches cannot be measured without such data. Moreover, environmental questions

8 | P a g e posed by governmental agencies and interested groups cannot be scientifically answered without such data. Figure 1: Location of the aquatic reserves surveyed in this study (circled) in Puget Sound. Base map source: WDNR (2011).

Efforts will be made to establish continuity with existing WDNR and WDFW data in an effort to define trends and develop an understanding of the conditions and processes affecting the study areas over time. To achieve this, the studies will use established standards and sampling methodologies developed and made available by WDFW. As the planned monitoring program is implemented over succeeding years, it will generate data that can be used to establish baseline conditions, define trends, document changes, and identify potential restoration opportunities. Monitoring within each reserve will be constrained by two primary factors: Human resources to conduct the monitoring and public access. In each reserve, interested citizens will be recruited and trained to conduct the Forage Fish monitoring. Table 1 lists the lead and associate organizations for each reserve that will seek recruits and coordinate training. Table 2 identifies the key individuals and their responsibilities.

9 | P a g e Public access is the second constraint to the monitoring work. In each reserve, beaches at or near public access points will be used for sampling. Additionally, permission may be sought to conduct sampling on tribal or private land, as necessary. The analysis of samples for identification of species and the volume of eggs requires scientific expertise that is beyond the capabilities of this citizen science program. Therefore, the samples will be fully documented and then turned over to WDNR or WDFW for appropriate laboratory analysis. An appropriate chain of custody will be maintained and documented throughout this process. Organization and Schedule Washington Environmental Council (WEC) is the coordinator for this program. This monitoring is being conducted in three aquatic reserves, each with its own stewardship committee and associated organizations. The Statement of Work (SOW, 2013) describes the structure as “Washington Environmental Council, with the project partners, will train the citizen committee members using established protocols and will work with the resource agencies to add value to their existing research and monitoring programs…” Table 1 shows the partners involved in this project. These partners will be responsible for all aspects of intertidal forage fish spawning monitoring including recruitment of volunteers, coordinate training in association with WDNR, provision and distribution of equipment, coordinate and supervise sample collection, review completed data forms, catalogue and safeguard collected materials, and coordinate the processing of the samples through WDNR in association with WDFW. Results from the laboratory analysis will be made available to the WEC and the partner organizations for inclusion in their reports. Table 2 lists the key individuals and their responsibilities in this project. Table 3 shows the schedule for this program under this program. It is anticipated that work will continue under the guidance of the stewardship committees and partner organizations. The WEC will assist in the standardization of protocols across the reserves as well as assist in communications and coordination with local and state agencies. Table 1: Lead organizations for organizing activities in each reserve Reserve Local organization(s) Fidalgo Bay Puget Sound Corps (PSC); Beach Watchers Nisqually Reach PSC, NRCSC, Nisqually Reach Nature Center (NRNC) Smith and Minor PSC, Whidbey Watershed Stewards Islands

Table 2: Key individuals and their responsibilities Individual Responsibilities Dion Jamieson Coordinate activities between PSC and ARCSC

10 | P a g e (WDNR) Pete Haase Coordinate collection in FBAR Yvonne Shevalier Coordinate collection in NRAR Robin Clark Coordinate collection in SMAR Project schedule The sampling program requires adequate training of the citizen scientists. Once collection is started, it will be performed monthly June-November and twice monthly December. If the survey continues beyond this survey year, monthly data collection will begin in March and twice monthly surveys will be conducted December-February. The schedule is shown in Table 3. Additional sampling may occur if resources are available. Table 3: Schedule of important milestones in this project Activity Date Recruitment of volunteers February-May, 2013 Training of sampling procedures April-May, 2013 Begin Monthly sampling: June, 2013 Begin Semi-monthly sampling December 2013 Publication of draft report for comment by Ecology’s NEP QA Coordinator and November 2013 peers Publication of final report December 31, 2013 Training Each partner organization will coordinate training events with the Washington State Department of Natural Resources Aquatic Reserves' Program. In some cases, The PSC Team will be available for additional on- the- job training for those citizen science volunteers wishing to expand their knowledge and understanding of the monitoring protocol. Online resources from WDFW are also available, and all citizen-science volunteers will be directed to these resources. These include forage fish beach survey training materials and established protocols, available at http://wdfw.wa.gov/conservation/research/projects/marine_beach_spawning/ (accessed March 26, 2013). All volunteers will also receive training in safe operations during collection and transport of samples. Additional training will include methods to avoid the transport and introduction of invasive species.

11 | P a g e Budget and funding This program is primarily a volunteer-run project and therefore does not incur costs for personnel. Only minor equipment costs are anticipated, and funding for these purchases is available through this grant, as stated in the SOW (SOW, 2013). Volunteer hours are used as indirect matching to grant funding. Staff support, including coordinating activities between reserves and preparation of reports, are provided through this grant, including general activities such as development of the stewardship committees and evaluating the implementation of the management plans that are beyond the scope of this QAPP. Quality Objectives The overall quality objective for this project is to provide high quality data on forage fish usage of three Washington State aquatic reserves. This QAPP describes how samples will be collected, along with ancillary data. Identification and enumeration of fish species is described in WDFW FF-03 Protocol (WDFW, 2011). The quality objectives for the field methods herein will be evaluated using indicators of precision, accuracy/bias, representativeness, comparability, completeness, and sensitivity (as applicable). The sampling locations and tidal elevations where sampling will occur at each reserve will be chosen to be representative of where various species of forage fish are expected to spawn. To achieve comparability over time, samples will be collected, handled, stored, and transported following standard protocols. The target for completeness is to successfully collect beach samples for 95% of the planned bimonthly and monthly sampling events. Precision, accuracy/bias, and sensitivity apply mainly to measurement of location using hand-held GPS units. To validate accuracy and insure consistency among units, the make and model of the GPS unit will be recorded as well as the specifications of the accuracy of the unit. The datum for each unit will be set to NAD 83 and positions recorded in decimal degrees. The unit will be calibrated vs. a known reference point at the start of each sampling event. Sampling coordinates are expected to be accurate and repeatable to within 3 meters. However, because of the dynamics of the beach environment, replicates surveys at the same location will not be attempted. Therefore, the GPS coordinates will be used for generalized mapping and not relocating transect locations. Additionally, photographs of each sampling transect line will be taken to both help identify the location of the survey and to provide a method to correct any errant GPS readings. Sampling Design and Procedures The sampling design follows the WDFW Intertidal Forage Fish Spawning Habitat Survey Protocols, Procedures for Obtaining Bulk Beach Substrate Samples (Penttila, 2011). Slight modification of the protocol has been made in steps 9

12 | P a g e and 10 to eliminate the need to use preservatives when possible. The protocol is as follows: Note: Sampling should occur on the lowest tide practicable. Prior to sampling any site consult tide tables to ensure you will be able to access the +7-9 (surf smelt) and +5-8 (sand lance) tidal height. It may also be necessary to obtain permission to access the beach from private or corporate landowners. Procedure: 1. Upon arriving on the beach, fill out the header information on the attached data sheet. Do not fill in “Reviewed by.” Before conducting the first sample, describe the character of the upland and beach environment using the codes provided on the back of the data sheet. For additional details on sample codes see Moulton and Penttila (2001). 2. Identify a landmark from which you will measure the distance to the bulk substrate sample tidal elevation. Typical landmarks include the upland toe of the beach, the last high tide mark or wrack line, and the edge of the water. 3. Measure the distance from the landmark to the tidal elevation to be surveyed. 4. Stretch a measuring tape at least 100 feet along the selected tidal height. Note that beach contours may cause the landmark to be “wavy” and that the tape should remain a consistent distance from the landmark. 5. Standing at the 50 foot mark of the measuring tape, record a GPS fix on the data sheet. 6. Using a 16-ounce sample jar or large scoop remove surface substrates to the maximum depth possible between 5cm and 10cm (2-4 in) from the location recorded in Step 5 above. Place the sediment in an 8 inch x 24 inch polyethylene bag or large, sturdy ziplock. You may need to take two scoops to get sufficient sediment, depending on the coarseness of the beach. 7. Walk ten paces (single steps) along the measuring tape, repeat the sediment scooping action, and place the sediment in the bag. Move an additional ten paces and repeat. Move an additional ten paces, approximately to the end of the tape, and repeat. The bag should now have sediment from four locations along the tape and be at least ½ to ⅔ full. Place completed waterproof sample label in bag with sediment. Label should include reserve name, Beach# sample#, data and sampler initials. 8. If additional transects, representing various tidal heights, along the beach are to be surveyed, place the sample bag in a cool, shady place and repeat the above procedures at these additional locations. If no additional samples will be taken, move on to wet sieving and winnowing the sample as described in the companion protocol Procedures for recovering winnowed light fractions subsamples of forage fish egg-sized material from bulk beach substrate samples. 9. Place each winnowed subsample in a sample jar, making sure threads are clean to ensure a tight seal. Label each jar (in pencil) with reserve name, date, and

13 | P a g e beach# sample#. If samples may not be transported within 24hours, the preservative Stockard’s Solution may be added. If it is added, the closed jar should be inverted at least three times to be sure all the preservative is distributed. When using Stockard’s Solution, mark the jar with a large X to indicate a preservative has been used. See Figure 2 as an example of labeling.

10. Store sample collections in a cool location, making sure the samples are not frozen. Transport samples to WDFW personnel for analysis within 24 hours of collection unless a preservative has been used. 11. If you have a camera, take several photos of the survey area showing sampling locations. Be sure to take photos from several perspectives (i.e., both up and down, as well as along, the beach). For each photo, record the cardinal direction you are facing on the data sheet in the comments field. For our purposes, one photo of the transect from either end is sufficient, but highly encouraged! Still important to record cardinal direction. The complete protocol, along with a list of required equipment, supplies, data form, and data form instructions are shown in Appendix A. Invasive species transport prevention All sample jars will be sealed prior to leaving the beach. All sampling equipment will be inspected for any substrate, fauna, or flora. Potentially contaminated equipment will be rinsed in seawater and wiped down until all potential contaminants are removed. Paper or cloth towels used will be bagged and either properly disposed of or decontaminated. Training will also include instructions for participants to inspect their clothing for any possible contamination and the need to decontaminate the clothing before returning to any marine area. All clothing and equipment should be thoroughly dry prior to returning to a beach. Observers should not wear felt-soled boots or shoes as these can’t be effectively decontaminated. Details and options for decontamination are given at http://www.ecy.wa.gov/programs/eap/InvasiveSpecies/AIS-PublicVersion.html. Standard laboratory precautions will be used to prevent any release of invasive species from the samples. Quality Control and Data Management The following procedures will be used to maintain quality of the samples and data in this program. 1. Follow the standard procedures as described in the previous section among sites and among monitors. This includes filling out the field form completely. 2. Only individuals who successfully completed training in the collection protocols are allowed to collect samples. Other individuals may participate in other non-collection activities, such as assisting with the

14 | P a g e establishment of a transect line, but only if they are accompanied and supervised by a trained individual. 3. A single individual, who has been successfully trained in the protocol, within the monitoring team, is responsible to the integrity of the sampling. This individual will be identified on the data collection form by being the first name entered under “Samplers.” 4. Sampling locations will be recorded from hand-held GPS units. The make and model of the units will be recorded. The datum for each unit will be set to NAD 83 and positions recorded in decimal degrees. The unit will be calibrated vs. a known reference point at the start of each sampling event. Sampling coordinates are expected to be accurate and repeatable to within 3 meters. 5. The container used to hold the samples will be labeled in pencil. This label should include: reserve abbreviation (FB, NR, or SM) date and beach number/sample number (B#S#) in pencil. 6. Prior to leaving each sampling area, the project manager/field lead will review field logs and other field notes to identify any unusual or anomalous results. If any are found, they will be resolved (e.g., repeat GPS measurements or sample collections, if necessary) before abandoning the field efforts. 7. Data will be entered into a WDNR database with the necessary oversight/error checking to ensure accuracy of transcription. Data deemed accurate and complete by WDNR will by input into WDFW’s Forage Fish database by the PSC. The WDFW database is viewable on SalmonScape at the following link, http://wdfw.wa.gov/mapping/salmonscape/index.html. Audits and Reports No Audits of this project are anticipated. A draft report containing the elements below will be prepared and distributed to Ecology’s NEP Quality Coordinator and peers, including WDFW, WDNR, and members of the Forage Fish work group of the Puget Sound Ecosystem Monitoring Program for comment. The final monitoring report will be completed by December 31, 2013. Production of the reports will be a collaborative effort between the WEC, the partner organizations, and WDNR. The final report will include:  A narrative of the field research  A summary of sampling effort  Analysis of the presence or absence of eggs by zone for those samples where laboratory analysis is complete.  An assessment of the feasibility of this study

15 | P a g e  Recommendations for any modification of the procedures and the overall program The final report will be available from the WDNR AR website at http://www.dnr.wa.gov/ResearchScience/Topics/AquaticHabitats/Pages/aqr_rsv e_aquatic_reserves_program.aspx References Liedtke, Theresa, Gibson, Caroline, Lowry, Dayv, and Fagergren, Duane, eds., 2013, Conservation and Ecology of Marine Forage Fishes—Proceedings of a Research Symposium, September 2012: U.S. Geological Survey Open- File Report 2013-1035, 24 p. http://www.nwstraits.org/uploads/pdf/NWS- 2013_FF_Symposium.pdf (accessed March 26, 2013) Moulton, Lawrence L and Penttila, Daniel E. 2001 (Revised 2006). San Juan County Forage Fish Assessment Project Forage Fish Spawning Distribution in San Juan County and Protocols for Sampling Intertidal and Nearshore Regions Final Report June 2000. Available at http://www.nwstraits.org/Uploads/pdf/sjc_forage_protocols.pdf (accessed April 4, 2013). Penttila, Dan. 2011. WDFW Intertidal Forage Fish Spawning Habitat Survey Protocols; Procedures for obtaining bulk beach substrate samples. WDFW Protocol FF-01 Version 1.0, July 2011. Reformatted by Dayv Lowry and available on request from Dayv Lowry at WDFW ([email protected]). Pierce, Ken; Penttila, Dan; Benson, Brian; Krueger, Kirk; Quinn, Timothy; and Price, David 2012. Spatiotemporal Detection of Forage Fish Eggs Derived from Long-term Spawning Surveys. Poster available at http://wdfw.wa.gov/publications/01211/wdfw01211.pdf (accessed March 26, 2013) Puget Sound Partnership. 2012: The 2012-13 Action Agenda for Puget Sound. (page 134) Available at: http://www.psp.wa.gov/downloads/AA2011/083012_final/Action%20Agenda %20Book%202_Aug%2029%202012.pdf (accessed March 26, 2013) SOW, 2013. SOW #12-1103. Puget Sound Marine and Nearshore Protection and Restoration Grant Program, Ensuring Regulatory Effectiveness in Puget Sound’s Most Special Places., Grant # PC-00J29801-0. WDFW, 1998. Forage Fish Management Policy, Goals and Plan. http://wdfw.wa.gov/commission/policies/c3012.html (accessed March 26, 2013) WDFW, 2011. Protocol FF-03 Intertidal Forage Fish Spawning Habitat Survey Protocols Laboratory procedure for determining forage fish egg presence/absence from preserved “winnowed light fraction” beach substrate samples. Available at http://wdfw.wa.gov/conservation/research/projects/marine_beach_spawning/trai ning/protocol-lab_spawn_presence_absence.pdf (accessed June 14, 2013)

16 | P a g e WDNR, 2008. Fidalgo Bay Environmental Aquatic Reserve Management Plan. http://www.dnr.wa.gov/Publications/aqr_rsve_fid_mgmt_plan.pdf (accessed March 26, 2013) WDNR, 2011. Nisqually Reach Aquatic Reserve Management Plan. http://www.dnr.wa.gov/Publications/aqr_nisqually_reserve_finalplan_2011.p df (accessed March 27, 2013) Appendix A: Protocols and Data Forms Collection Protocol and Data Recording Form See supporting documents Forage Fish Spawning Survey Quantitative Protocol for sample collection protocol, data form, and recording codes and see Sample Completed Data Form for an example of a properly completed data form. Condensing Bulk Samples Protocol The following protocol for “winnowing” the samples to remove excess material and collect the lighter fraction that may contain eggs is derived from Moulton and Penttila, (2000). Equipment • Rack of sediment screens, size 4, 2, and 0.5 mm, preferably Nalgene instead of the more traditional brass screens, • 2 - 5 gallon buckets modified to act as drain for screen rack, • Wash bucket, • Plastic dishpan, • 16 ounce sample jar • Bristle brush for cleaning screens between samples Procedure The bulk egg samples can be processed in the field to remove most of the sand and reduce the volume of the sample. This is done by washing the eggs from the sand and discarding the barren sediment. The eggs are lighter than the sand and gravel and will move upward during the washing process, allowing them to be skimmed from the surface of the material. The washing is conducted as follows: • Assemble the screens on top of the drain bucket, with the largest mesh on top, grading to the smallest mesh on the bottom. • Remove the sample label and place it in an 16 ounce sample jar. • Add the sample to the top screen; thoroughly wash the sediment through the screen set with either salt or fresh water, whichever is readily available. If the

17 | P a g e sample is large you may have to split in and repeat the following steps with the remaining half. • Discard the sediment in the top screens; retain only the material in the bottom (0.5 mm) screen. • Dump the material retained in the 0.5 mm screen into the dishpan. • Add water until the material is covered by 1-2 inches of water. • Swirl the water around the pan, adding rocking and bouncing motions to allow the eggs to migrate to the top of the sediment. The idea is similar to gold panning, try to winnow the eggs to the surface of the material. • After swirling for 1-2 minutes, work the lighter fraction of material to one corner of the pan.  Carefully dry up the lighter fraction by tipping the pan so that the water drains away, and skim the lighter fraction from the surface of the sand with the sample jar. • Repeat the winnowing process 2-3 more times, each time adding the retained lighter fraction to the sample jar, until you are left with a sample jar ½ to 2/3 full. • Gently rinse the threads of the jar with the sample water to ensure a tight seal.

Appendix B: Glossary, Acronyms, and Abbreviations Glossary—Quality Assurance Accuracy - the degree to which a measured value agrees with the true value of the measured property. USEPA recommends that this term not be used, and that the terms precision and bias be used to convey the information associated with the term accuracy. (USGS, 1998) Bias - The difference between the population mean and the true value. Bias usually describes a systematic difference reproducible over time, and is characteristic of both the measurement system, and the analyte(s) being measured. Bias is a commonly used data quality indicator (DQI). (Kammin, 2010; Ecology, 2004) Calibration - The process of establishing the relationship between the response of a measurement system and the concentration of the parameter being measured. (Ecology, 2004) Comparability - The degree to which different methods, data sets and/or decisions agree or can be represented as similar; a data quality indicator. (USEPA, 1997)

18 | P a g e Completeness - The amount of valid data obtained from a data collection project compared to the planned amount. Completeness is usually expressed as a percentage. A data quality indicator. (USEPA, 1997) Precision - The extent of random variability among replicate measurements of the same property; a data quality indicator. (USGS, 1998) Quality Assurance (QA) - A set of activities designed to establish and document the reliability and usability of measurement data. (Kammin, 2010) Quality Assurance Project Plan (QAPP) - A document that describes the objectives of a project, and the processes and activities necessary to develop data that will support those objectives. (Kammin, 2010; Ecology, 2004) Sample (field) – A portion of a population (environmental entity) that is measured and assumed to represent the entire population. (USGS, 1998) Sensitivity - In general, denotes the rate at which the analytical response (e.g., absorbance, volume, meter reading) varies with the concentration of the parameter being determined. In a specialized sense, it has the same meaning as the detection limit. (Ecology, 2004) Standard Operating Procedure (SOP) – A document which describes in detail a reproducible and repeatable organized activity. (Kammin, 2010) References Ecology, 2004. Guidance for the Preparation of Quality Assurance Project Plans for Environmental Studies. http://www.ecy.wa.gov/biblio/0403030.html USEPA, 1997. Glossary of Quality Assurance Terms and Related Acronyms. http://www.ecy.wa.gov/programs/eap/qa.html Kammin, 2010. Definition developed or extensively edited by William Kammin, 2010. USGS, 1998. Principles and Practices for Quality Assurance and Quality Control. Open-File Report 98-636. http://ma.water.usgs.gov/fhwa/products/ofr98-636.pdf

Glossary – General Terms Stockard’s Solution: A formalin based preservative that also include acetic acid, glycerin, and water (Moulton and Penttila, 2001).

Acronyms and Abbreviations Following are acronyms and abbreviations used frequently in this report. AR Aquatic Reserve ARCSC Aquatic Reserve Citizen Stewardship Committee CSC Citizen Stewardship Committee e.g. For example Ecology Washington State Department of Ecology EPA U.S. Environmental Protection Agency

19 | P a g e et al. And others FB Fidalgo Bay FBAR Fidalgo Bay Aquatic Reserve GPS Global Positioning System i.e. In other words NOAA National Oceanographic and Atmospheric Administration NR Nisqually Reach NRAR Nisqually Reach Aquatic Reserve NRNC Nisqually Reach Nature Center PI Principal Investigator PSC Puget Sound Corps PSP Puget Sound Partnership QA Quality assurance QAPP Quality Assurance Project Plan SM Smith and Minor SMAR Smith and Minor Islands Aquatic Reserve SOP Standard operating procedures SOW Statement of Work WDFWWashington Department of Fish and Wildlife WDNR Washington Department of Natural Resources WEC Washington Environmental Council Units of Measurement ft feet km kilometer, a unit of length equal to 1,000 meters. m meter mm millimeter

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