ABSTRACTS 68th Annual Pacific Coast Shellfish Growers/ Pacific Coast Section National Shellfisheries Association Conference and Tradeshow Vancouver, WA, September 22-25, 2014 PUTTING SHELLS BACK IN THE WATER: UNDERSTANDING SHELL DISSOLUTION FOR ALKALINITY REGENERATION ABDULGHANI, Lisa*, and Erin MARTIN, The Evergreen State College, Olympia, Washington Spreading shell material can buffer corrosive conditions by providing alkalinity regeneration by dissolution of calcium carbonate (CaCO3). This research explored how enhancing the seafloor with a particular size or species of bivalve shell may influence different rates of shell degradation in Hood Canal, Washington. The differences in degradation, measured by changes in mass, were examined over an incubation period of eight weeks among whole and crushed size types for three different species: Crassostrea gigas, Ostrea lurida, and Mytilus galloprovincialis. All shell treatments lost mass, while M. galloprovincialis shells degraded the most mass, losing up to 2.78% ± 0.08% of its shell matter. Each species had a significantly different rate of mass loss relative to the other species, whether the shell was crushed (F2,87 = 37.39, p ADAPTATION OF RESTORATION PRACTICE: NATIVE OYSTER HABITAT AND POPULATION METRICS PROVIDE FEEDBACK AND GUIDANCE FOR ENHANCEMENT TECHNIQUES ALLEN, Brian *, Puget Sound Restoration Fund Since 2010, PSRF has been collecting displacement volume measurements from samples of emergent oyster substrate collected within native oyster (O. lurida) restoration project sites and natural aggregations in Puget Sound. This method for quantifying substrate "amount" has allowed an analysis that includes shell application rate and resulting habitat amounts with population parameters of interest for restoration (e.g. recruitment magnitude, adult density). We use two case studies from Kitsap county to develop an application scheme with experimental utility for a 2014/2015 habitat and stock rebuilding project over large spacial scales. 2 DEVELOPMENT OF THIRD-GENERATION, HIGH DENSITY, LINKAGE MAPS FOR THE PACIFIC OYSTER ARIAS-PEREZ, Alberto*, University of A Coruna and University of Southern California HEDGECOCK , Dennis , University of Southern California Linkage maps are a fundamental genomic tool to study performance and production traits. Nevertheless, for most aquaculture species the density of available maps has limited their utility. For the Pacific oyster, different technologies and markers (AFLPs, microsatellites and recently single-nucleotide polymorphisms, SNPs, typed using the Illumina GoldenGate Genotyping Assay) have been employed to build linkage maps. However, these approaches are labor intensive, expensive and yield a limited number of markers. Recent advances in sequencing technologies have made it possible to discover and genotype simultaneously thousands to tens of thousands of SNPs in a cost-effective way. To increase marker density we employed a protocol that uses a restriction enzyme to target a subset of the genome and barcoded adapters to allow sequencing multiple individuals at the same time. After removing potential variants with low coverage, individuals with low number of sequences, nonsegregating variants and unexpected genotypes a preliminary analysis of the data identified, for the two parents and 93 offspring of a full-sib family, 3664 SNP markers. When these SNPs and 37 microsatellite markers were used to build a linkage map, ten main linkage groups were observed comprising 1442 markers, spanning 874.5 cM, with an average spacing of 0.8 cM. As observed with SNPs from protein-coding sequences, some markers that align to the same genomic scaffold were assigned to different linkage groups, suggesting miss-assembling of scaffolds. The markers identified in this work will help to improve the assembly of the oyster genome and to build third-generation linkage maps that will be useful to study quantitative traits. 3 DIETARY LIPID AFFECTS THE NUTRITION AND CONDITION OF RED KING CRAB LARVAE (Paralithodes camtschaticus) BEDER, Asia *, School of Fisheries and Ocean Sciences, Juneau Center, University of Alaska Fairbanks, Juneau, Alaska, USA. COPEMAN, Louise , College of Earth, Ocean, and Atmospheric Sciences, Oregon State University, Corvallis, OR, USA and Cooperative Institute for Marine Resources Studies, Oregon State University, Hatfield Marine Science Center, Newport, Oregon, USA. ECKERT, Ginny L., School of Fisheries and Ocean Sciences, Juneau Center, University of Alaska Fairbanks, Juneau, Alaska, USA. Six of nine red king crab stocks in Alaska crashed, the fisheries are closed, and stocks have not recovered. The Alaska King Crab Research Rehabilitation and Biology (AKCRRAB) program is investigating the feasibility of enhancement to rehabilitate these stocks. The goal of this research project is to improve larval survival through improved nutrition and therefore increase hatchery larval production. We raised larvae from eight ovigerous red king crab females in twelve 190 L tanks at the Alutiiq Pride Shellfish Hatchery, in Seward, Alaska and varied the nutritional content of the larval diet by feeding larvae Artemia nauplii enriched with lipid emulsions varying in essential fatty acids (EFA): 1) high DHA, 2) high DHA and EPA, 3) high DHA and AA, and 4) a control lipid (low EFA). We found similar larval survival to glaucothoe among the diet treatments. Larval condition during the four zoeal stages (assessed by viewing size and number of lipid droplets) generally did not vary among diets; however, the glaucothoe fed diets enriched with high DHA and EPA and high DHA and AA had more and larger lipid droplets than the DHA and control lipid diets. During a stress test that measured response time after exposure to freshwater, glaucothoe that were fed the diet enriched with the control lipid (low EFA) recovered significantly slower then the other diets, suggesting that EFA can improve performance during stress. We also investigated differences in lipid classes and fatty acids (FAs) among the larvae fed the four different diets during the fourth zoeal and glaucothoe stages. We found significant differences in the proportions of EFAs in the larvae during both stages. Specifically, EPA proportions varied between all the diets during both stages. Proportions of AA were significantly higher in crabs fed the DHA and AA diet during both stages. Also, proportions of DHA were significantly higher in crabs fed just a DHA diet and the control diet crabs had significantly less EFA then crabs fed all other diets. During the glaucothoe stage, crabs receiving the control diet had significantly lower total fatty acids (mg/g-1 wwt). When comparing strong and weak crabs from the stress test we found no significant difference in FAs. Results from this study will be used to improve diets of hatchery-reared king crab to increase larval production. 4 IMPACT OF IMIDACLOPRID ON BENTHIC INVERTEBRATES ON LARGE PLOTS BOOTH, S.; SUHRBIER, A.; MIDDLETON, M.; HUDSON, B., Pacific Shellfish Institute, Olympia WA PATTEN, K., Washington State University, Long Beach, WA BARRET, J. These studies were conducted in response to Washington Department of Ecology’s request for further descriptions of the Sediment Impact Zone on large acreages of commercial tidelands in Willapa Bay treated with imidacloprid to control burrowing shrimp, as previous studies were conducted on isolated plots <10 ac in size. An initial proposal to WDFW was funded, but was modified and expanded after discussions with Ecology. Results from a similar study conducted in the Cedar River area in 2011 impacted the design of the 2014 studies. Ultimately, an 85 ac plot in the Stony Pt area and a xx ac plot near the Cedar River, where levels of silts are higher and tidal flushing is lower, were treated aerially with a flowable formulation of imidacloprid. Concentrations of imidacloprid in the pore water and sediments sampled by WSU at sites both on and off the plots at 1, 14, and 28 days after treatment are presented. Benthic and epibenthic invertebrates were sampled only at the Stony Pt plot and at two untreated plots of similar substrate and vegetation near Bay Center. Preliminary results from the benthic samples show substantial pre-treatment differences between the Stony Pt and Bay Center areas. Results of the 2011 Cedar River study will also be presented as time allows. 5 MICROORGANISMS ASSOCIATED WITH SEA STAR WASTING DISEASE BURGE*, Colleen A., School of Aquatic & Fishery Sciences, University of Washington, Box 355020, Seattle, WA HEWSON, Ian, B.M. GUDENKAUF, J. BUTTON, Department of Microbiology, Cornell University, Wing Hall, Ithaca, NY HARVELL, C. Drew, M. EISENLORD, Department of Ecology & Evolutionary Biology, Cornell University, Corson Hall, Ithaca, NY NEWTON, Alisa L., Wildlife Conservation Society, 2300 Southern Boulevard, Bronx, NY Pete RAIMONDI, M. MINER. Department of Ecology & Evolutionary Biology, University of California - Santa Cruz, Santa Cruz, CA MINER, B., Department of Biology, Western Washington University, 516 High Street, Bellingham, WA A large-scale mortality event affecting 20 species of sea star (Asteroidea) on the Pacific Coast of North America began in June of 2013 and continues to present (August 2014). First detected in Ochre sea stars (Pisaster ochraceus) along the outer coast of Washington State, the event has been observed in at least 250 subtidal and intertidal locations extending from Southeastern Alaska to San Diego, California. Citizen science has been a key approach to monitoring the spread of disease. Pisaster, Pycnopodia and Evasterias are the most affected genera and mortality approaches 100% in select species at some locations. Both Pisaster and Pycnopodia are considered keystone species, and changes to community composition within ecosystems are possible. The progression of clinical signs includes: 1) loss of body turgor (deflation) and weakness; 2) foci of epidermal pallor and tissue loss; 3) sloughing of multiple rays and/or rupture of the body wall with evisceration; 4) death. Multiple approaches, including microscopic (histology, electron microscopy, in situ hybridization), genomic (metagenomics, community finger-printing, and transcriptomics), culture, experimental (infection trials), and monitoring are currently underway. Experimental trials indicate an infectious agent is involved, and possible bacterial and viral agents have been identified. OLYMPIA OYSTER RESTORATION: HISTORY AND POLITICS CAMMER SULLIVAN, Nina*, The Evergreen State College My research was on native Olympia Oyster history and politics. Specifically the history of farming Olympia Oysters and it’s impact in policy making. During the 1920’s in Shelton, WA pulp and paper mills polluted Puget sound inlets harming Oyster cultivation. Chemical outputs at the 6 time from mills affected local economies, ecosystems and policies. Shellfish though delicious to dine on, are treated as a resource in Washington. Studies I did focused on native land rights in Washington surrounding Shellfish and policies. And what methods are being made today including cultivation to restore Olympia Oysters. THE SUSTAINABILITY OF THE CALIFORNIA SEA CUCUMBER (Parastichopus californicus) AND RED SEA URCHIN (Mesocentrotus franciscanus) DIVE FISHERIES IN THE SAN JUAN ISLANDS, USA CARSON, Henry S. *, Washington Department of Fish and Wildlife ULRICH, Michael, Washington Department of Fish and Wildlife The San Juan Archipelago is the most intensely fished region of Washington State for echinoderms, and dive fisheries for both the California sea cucumber (Parastichopus californicus) and red sea urchin (Mesocentrotus franciscanus) experienced overharvest in the late 1980s and early 1990s. Here we evaluate the current management of these fisheries using a biomass estimate from a remote-operated vehicle survey, a time series of relative abundance from SCUBA index station surveys, and harvester log book data. We also report habitat associations of both species with depth and seafloor substrate. The fully-utilized quota for Parastichopus represents an 11.4% harvest rate on the current harvestable biomass estimate, density in shallow water is conspicuously low, relative abundance has remained depressed, and catch-per-unit-effort (CPUE) continues to decline. Abundant animals below harvestable depths may not be of sufficient density to act as an efficient reservoir to replenish the shallows with recruits. The partially-utilized quota for Mesocentrotus represents a 3.9% harvest rate on the current biomass estimate, density in shallow water is conspicuously low, relative abundance is increasing from a recent low, and there is no trend in CPUE. Despite the challenges of comanaging the fisheries among several regions and stakeholder groups, agreement has been reached to improve the long term sustainability of the Parastichopus fishery using reduced harvest quotas and a closure during peak spawning months, and to continue to closely monitor the Mesocentrotus fishery. 7 INVESTIGATING ALTERNATIVE AGING METHODS FOR BURROWING SHRIMP POPULATIONS IN YAQUINA BAY COLEMAN, Natalie, Oregon State University, Department of Integrative Biology, Corvallis, Oregon BOSLEY, Katelyn, Oregon State University, Department of Fisheries and Wildlife, Hatfield Marine Science Center, Newport, Oregon DUMBAULD, Brett - US Department of Agriculture, Agricultural Research Service, , Hatfield Marine Science Center, Newport, Oregon The burrowing shrimp Neotrypaea californiensis is an important ecosystem engineer inhabiting estuaries along the Pacific Northwest coast. The burrowing action of these shrimp is a threat to oyster aquaculture, especially in Washington state where the topical pesticide carbaryl has been sprayed to control shrimp populations. More information is needed about this species, including a viable aging method to assist in developing an integrated pest management plan for these shrimp. This is difficult due to crustaceans molting their hard parts which may retain annual deposits, an accurate method used in aging other species. This study explores verification of two methods, lipofuscin analysis and gastric mill rings, on shrimp populations by using these methods on known-age shrimp. Counts of growth rings located in the pyloricossicle as well as the aging pigment lipofuscin were completed on each sample. Temperature data from the known-age experiment was also collected to compare growth and number of rings with temperature degree day. Lipofuscin values of known-age shrimp were similar while size varied, verifying that this method is useful in the direct aging of crustaceans. Number of rings was shown to correlate more directly with growth than with lipofuscin, suggesting that they might not be a direct measure of linear age but rather metabolic age, which is more affected by environmental factors than LF analysis. . Size was scattered across all age classes assigned which suggests size is not a reliable indicator of age in N californiensis but LF-based age classes can be determined which should enable creation of age structure and cohort based population models. PSI OUTREACH: CONVINCING FOLKS TO KEEP IT CLEAN! CHRISTY, Aimee* and Mary MIDDLETON, Pacific Shellfish Institute 8 POTENTIAL EFFECTS OF NON-NATIVE EELGRASS ON BAY CLAM POPULATIONS IN NETARTS BAY, OREGON D’ANDREA*, Anthony, PEROTTI, Elizabeth, MOFFETT, Cinamon, and STRICKLAND, Stacy Oregon Department of Fish and Wildlife, Marine Resources Program, Newport, OR The non-native eelgrass Zostera japonica was introduced in the 1930s to the Pacific Northwest and has since become well established in many estuaries. Several of these estuaries have expansive native Zostera marina beds and support large recreational and commercial shellfish species . However, there is limited information about the impacts of Z. japonica expansion on ecosystem structure and function. In 2013-2014, the Shellfish and Estuarine Assessment of Coastal Oregon (SEACOR) project of the Oregon Department of Fish and Wildlife conducted a bay clam population and estuarine habitat study in Netarts Bay. This estuary is a popular recreational clamming destination and also supports commercial mariculture operations. Both rapid and detailed assessment methods were applied extensively across the study area in grid and random stratified points with respect to tidal elevation. Post hoc, we identified four bed types in the bay (Z. marina, Z. japonica, mixed eelgrass species, and non-vegetated beds), and explored patterns in the related benthic invertebrate communities. Z. japonica is widely distributed in the bay with an overlapping distribution with Z. marina. We found a significant relationship between bed type and community composition. Additionally, some species of recreationally-targeted bay clams in the upper intertidal were significantly associated with bed types which included Z. japonica. Notably, native littleneck (Leukoma staminea) clams were more prevalent in mixed eelgrass beds. These survey results enhance our understanding of distribution and potential effects of Z. japonica but highlight the need for further research on the interactions of this non-native eelgrass species on bay clam populations. GETTING CREDIT FOR GROWING SHELLFISH DENIKE*, Jesse. G., and Peter H. DYKSTRA. Plauche & Carr LLP, Seattle, WA 98104. Shellfish create direct economic returns from harvest, and they are also a natural resource that provides a multitude of valuable ecosystem services. These services include water quality improvement from filter-feeding, de-nitrification of sediments, and enhanced fisheries productivity and shoreline stabilization from healthy shellfish reefs. Historically, these ecosystem services have not been recognized in the marketplace or in the regulatory environment. However, these services are increasingly being recognized and quantified by resource managers and regulators. Agencies that have traditionally been exclusively regulators are beginning to play a vital role in creating incentives to enhance ecosystem services. Doing so requires these agencies to work in partnership with regulated industries like shellfish farming. The regulated industries need these partnerships to achieve 9 the regulatory goals, and regulatory agencies can benefit from the innovation and investment private businesses can make in the tools to produce and measure ecosystem services. This presentation will explore ecosystem services, their role in solving complex regulatory challenges, and their potential for creating new market opportunities for shellfish farmers. It will include specific examples of potential avenues for shellfish farmers to realize returns for providing ecosystem services while discussing current regulatory and market obstacles to capturing the full value of these services. BREEDING OYSTERS FOR THE WEST COAST IN CHANGING OCEAN CONDITIONS: AN UPDATE ON THE COLLABORATIVE MOLLUSCAN BROODSTOCK PROGRAM (iMBP) DURLAND*, Evan, Chris LANGDON, Blaine SCHOOLFIELD, and Javan BAILEY, Coastal Oregon Marine Experiment Station, Oregon State University, Newport, OR 97365 The industry-supported Molluscan Broodstock Program (iMBP) focuses on improvement in survival, growth, yields and other desirable traits of West Coast cultured oyster species. In addition, the program acts as a repository for valuable stocks of Kumamoto and Midori oysters brought to the West Coast from southern Japan in 2006 and 2004, respectively. The program is partially supported by donations from industry members and builds on about 18 years of selective oyster breeding. iMBP has harvested cohorts 23 and 24 in 2013 and 2014, respectively. Results for cohort 23 planted inter-tidally in Netarts Bay, Oregon, (exposed to oceanic conditions) and Oyster Bay, Washingon, (exposed to high summer water temperatures) indicated that average yields of iMBP families were better than those of groups derived from wild broodstock. Yields of the same families planted in both sites were weakly correlated (r2 = 0.15), indicating that genotype x environment interaction effects on family yields were stronger than for many previous cohorts. In 2014, cohort 25 was produced from top-performing families of cohorts 22 and 23. In addition, pods and a mass spawning of the Midori stock were produced at commercial hatcheries for production of large numbers of broodstock. The future plans for iMBP will be discussed. 10 BURROWING SHRIMP RECRUITMENT TO COASTAL ESTUARIES AND SHELLFISH AQUACULTURE BEDS DUMBAULD*, Brett R., Lee McCOY, USDA-Agricultural Research Service, Hatfield Marine Science Center, Newport, OR 97365 BOSLEY, Katelyn, Department of Fisheries and Wildlife, Oregon State University, Hatfield Marine Science Center Newport, OR 97365 Two species of burrowing shrimp, Neotrypaea californiensis and Upogebia pugettensis are important members of intertidal mudflat communities in Pacific Northwest coastal estuaries but cause problems for shellfish farmers. We have monitored populations of these shrimp in Willapa Bay, Washington for two decades and in several other estuaries since 2005. Densities of both shrimp species were either increasing or stable through the mid 1990’s in Willapa Bay and then began to decline. U. pugettensis are now almost absent in Willapa Bay and many other estuaries along the West Coast due in part to the introduced parasitic isopod, Orthione griffenis which compromises their reproduction. Recent surveys of N. californiensis populations however, suggest that they too were declining. Since both shrimp have pelagic larval stages that develop in the coastal ocean, we asked whether interannual fluctuations in larval survival and estuary recruitment influenced adult populations and whether this influenced shrimp populations on shellfish aquaculture beds. Relatively high recruitment for both shrimp was observed in Willapa Bay through the mid 1990’s, but a period of about 10 years lapsed with very low recruitment. This loosely correlates with the decline in adult densities in that estuary and we also found a relationship between recruitment and the 1+ population. We observed a substantial ghost shrimp recruitment event to Yaquina Bay, Oregon in 2010 and repeated though slightly lower recruitment Willapa Bay from 2011- 2013, and are tracking survival and growth of these shrimp. This data provides further insight into the complexity of shrimp population dynamics. EELGRASS AND OYSTER AQUACULTURE INTERACTION AT THE LANDSCAPE SCALE IN WILLAPA BAY WASHINGTON DUMBAULD*, Brett R. and Lee McCOY, USDA-Agricultural Service, Hatfield Marine Science Center, Newport, OR 97365 Both seagrasses and bivalve shellfish provide valuable ecosystem services including estuarine nursery habitat in estuaries. Seagrasses are protected by no-net-loss provisions in federal and state regulations resulting in a precautionary approach by managers that avoids any direct impacts from development activity including shellfish aquaculture. Recent research suggests that oyster aquaculture has direct impacts to the native seagrass (eelgrass, Zostera marina) in U.S. West coast estuaries over small spatial and short temporal scales. We quantified the impacts of oyster aquaculture on Z. marina at the larger estuarine landscape scale in Willapa Bay, Washington which produces approximately 17% of US oysters. A model of Z. marina 11 density outside of aquaculture was first created using distance to mouth, distance to channel, salinity, elevation, and cumulative wave stress as predictors. We then used these factors to predict Z. marina distribution for aquaculture beds. We found the amount of Z marina observed on oyster aquaculture beds was less than the amount predicted by the model, but compared to the total amount of Z. marina in Willapa Bay, the impact of oyster aquaculture represented less than 1.5% of the total predicted values in any year. Type of oyster culture bed (longline, seed , and mixed) did not contribute to explaining observed variation, but mechanically harvested beds had significantly lower Z. marina than hand and mixed harvest beds. Nonetheless, the majority of beds (all harvest types) had 65-145% of the model predicted Z. marina and exhibited relatively low variability between years, suggesting that Z marina as habitat is resilient and that oyster aquaculture does not cause persistent effects at the landscape scale in this estuary. UPDATE - SHELLFISH DISEASES AND REGULATIONS ELSTON, Ralph A., AquaTechnics, Inc., Carlsborg, Washington 98324 Interstate movement of shellfish to receiving waters requires a permit from the receiving state, provincial or national jurisdiction. To address both the interstate movement of shellfish and export, a voluntary Shellfish High Health Program was developed in 2000 for seed producers and exporters. In late 2012, regulations for moving live shellfish from the United States into Canada were federalized by the Canadian government. The status of this ongoing action will be updated. Perkinsus marinus and P. olseni are reportable bivalve mollusk diseases not present on the west coast of the United States. Particular states enjoy freedom from additional shellfish diseases. For example, Hawaii and Alaska are free of Denman Island disease. Maintenance of disease freedom status increases the potential for export and decreases the losses to and cost of management of edible shellfish. Denman Island disease is caused by the protozoan parasite Mikrocytos mackini, first discovered in Pacific oysters (Crassostrea gigas) near Denman Island in British Columbia in the 1960s. Mikrocytos mackini was delisted by the World Animal Health Organization (OIE) as an internationally reportable disease in 2007. Prior findings reported in 2011 suggested that Kumamoto oysters are more susceptible to Mikrocytos mackini (causative agent of Denman Island disease) than Pacific oysters. An update on the status and knowledge of M. mackini in Kumamoto oysters will be presented. Finally, Pacific oyster seed production was persistently affected by vibrio infection in 2014, suggesting that environmental factors in seed production areas favored these outbreaks of vibriosis. 12 Alexandrium CYST SURVIVAL IN THE SEDIMENTS FEIFEL, Kirsten *, Washington State Department of Natural Resources, Olympia, WA, and University of Washington, Seattle, WA . L. WATSON, L., S. FLETCHER , and E. LESSARD, University of Washington, Seattle, WA Alexandrium is a common temperate dinoflagellate that is known to produce a suite of toxins, causing paralytic shellfish poisoning (PSP), if consumed by humans. For most of the year, Alexandrium cells lie dormant on the sediment floor in a quiescent stage known as a cyst. Cysts on the seafloor provide the inoculum for annual blooms. The preservation potential, survival and/or degradation of Alexandrium cysts in the sediments are not well-understood. It has been presumed that Alexandrium cysts have a relatively short lifetime in the sediments but recent work has shown that cysts can remain viable for up to 100 years. To investigate the percent survival and germination capability of Alexandrium cysts in the natural environment, we isolated over 300 cysts from a 60-cm sediment core extracted from Sequim Bay, WA in 2012. The sediment core was dated using the naturally occurring isotope, Lead-210, and was estimated to cover an almost 80 year period. We hypothesized that older Alexandrium cysts, isolated from deeper in the sediment core, would have a reduced ability to germinate when placed under "ideal growth conditions" relative to younger cysts isolated from surface sediments. Results indicated that on average, cysts isolated from deeper in the sediment core are just as likely to germinate as are cysts isolated from the surface of the sediment core. We found no evidence of cyst deterioration or reduced viability over the 80-year timescale considered in this study. A NOVEL TECHNIQUE FOR EVALUATING THE FEEDING PHYSIOLOGY AND BEHAVIOR OF BIVALVE LARVAE GRAY, Matthew*, and CHRIS LANGDON, Coastal Oregon Marine Experimental Station, Hatfield Marine Science Center, Oregon State University, Newport, Oregon 97365. KRAMER, Sean, Dept. of Mathematics, College of Science & Mathematics, Norwich University, Northfield VT 05663 Historically, feeding studies with bivalve larvae have indirectly measured feeding activity by measuring the decrease in concentrations of suspended algal cells using electronic particle counters; however, this method does not take into account production of fecal and pseudofecal particles that may be difficult to separate from uneaten algal cells, resulting in an underestimation of feeding rates. Using a series of exposures to differently colored fluorescent polystyrene microspheres, we describe a novel technique to directly quantify ingestion rates and gut fullness, and indirectly determine clearance rates and gut passage time of mussel larvae. Additionally, microsphere processing patterns observed using this method provided insight into digestive kinematics of the larval gut. In this study, we tested underlying 13 assumptions of the method and its effectiveness for evaluating the impact of development and temperature on the feeding activity of Mytilus galloprovincialis larvae. We conclude this technique is a powerful new tool for evaluating the feeding and digestive processes of bivalve larvae and potentially other planktotrophic invertebrate larvae. INVESTIGATING LONG-TERM CHANGE IN INTERTIDAL INVERTEBRATE POPULATIONS AT KUKUTALI PRESERVE, PUGET SOUND, WASHINGTON GREINER*, Courtney, and Julie S. BARBER*, Swinomish Indian Tribal Community, Department of Fisheries, La Conner, WA 98257 HOUGHTON, Jonathan P. , University of Washington, College of Fisheries, Seattle, WA 98105. Current address: Hart Crowser, Inc. Long-term research provides valuable insight into ecological processes and environmental change. Results from decadal studies are especially informative when assessing human impacts, whether direct or indirect. From 1969-1971 an intensive intertidal ecology study was conducted at Kukutali Preserve (KP), near La Conner, WA, an area that had historically served as an important bivalve digging zone. The Preserve became a de facto no-take marine reserve two decades after the study, providing an opportunity to replicate the investigation following cessation in harvest activities. For this component of the project we addressed the following questions (1) do targeted and non-targeted species densities change by elevation and year, and (2) do targeted and non-targeted species densities or biomass change by beach aspect (i.e. north, south) and year. Over four decades, KP experienced a significant decline in four of the seven species examined in this study. The only species that increased in density/biomass was the targeted butter clam, Saxidomus gigantea. The four species that declined in density/biomass included target (Leukoma staminea) and non-target species (Littorina sitkana, Littornia scutulata, and Hemigrapsus nudus). Regardless of other variables, we detected no change in two non-target species, Hemigrapsus oregonensis and Macoma inquinata. In general, the northern beaches supported heavier biomass measurements of bivalves and, as expected, most species densities varied with respect to elevation. Because the original study was not designed for multivariate analyses, our ability to determine the exact causes for these results was limited. The lack of harvest pressure cannot explain these outcomes, however, since harvest cessation typically results in an increase in target species density and biomass and our data did not correspond with this premise. Thus, we qualitatively considered pollution, predator-prey interactions, temperature, pH, and shoreline development as possible reasons behind the changes. 14 NATIVE OYSTER STOCKS FOR RESTORATION: DOES LOCAL ADAPTATION AFFECT Ostrea lurida OUTPLANTING? HEARE, Jake, Brent VADOPALAS, Joth DAVIS, and Steven ROBERTS, University of Washington, Seattle, Washington 98195 Olympia oysters, Ostrea lurida, in Puget Sound, Washington are known to initiate reproduction at a specific temperature threshold. Bays along a latitudinal gradient within the Sound exhibit temporal variation in attaining this temperature. This gradient of habitat types has been shown in recent studies (Savolainen, 2007) to induce the phenomena of local adaptation within semi sessile and sessile native organisms. Since O. lurida is native to the west coast, it is hypothesized that populations along a latitudinal gradient have become locally adapted to their environment. This adaptation would have important ramifications for conservation and restoration projects within the Puget Sound. To test these differences we set up a reciprocal transplant experiment among three populations from Fidalgo (Northern), Dabob (Central), and Oyster (Southern) bays along a latitudinal axis as well as a fourth bay (Manchester NOAA facility) as a control repository. We monitored growth, survival, and fecundity from August 2013 to August 2014. We observed only minor growth and survival differences among populations. Throughout the spawning season, Southern oyster population produced significantly more brooding oysters at two of the three sites compared to the other two populations. Oyster populations native to these diverse bays may have genetically diverged their spawn timing to conform to environmental conditions within each bay, or the Southern population may simply have greater fitness. Through our ongoing research, we intend to determine whether Olympia oyster populations exhibit local adaptation within Puget Sound. SECOND-GENERATION LINKAGE MAPS REVEAL ERRORS IN THE ASSEMBLY OF THE PACIFIC OYSTER GENOME AND PERMIT MAPPING OF THE GENETIC BASES OF PRODUCTION TRAITS HEDGECOCK, Dennis*, University of Southern California. Linkage maps are used to locate regions of the genome that influence production traits, such as survival, sex determination, and growth, of the Pacific oyster. With the recently published Pacific oyster genome (Zhang et al 2012 Nature), we should soon be able to dissect these regions, to determine the specific genes underlying variation in production traits. Initially, linkage maps were constructed with microsatellite DNA markers (Hubert & Hedgecock 2004 Genetics), which yielded ten linkage groups, corresponding to the number of chromosomes, but with fairly sparse coverage. To improve coverage and candidate gene-discovery, we developed 1536 single-nucleotide polymorphisms (SNPs), from protein-coding sequences, and typed these in five full-sib families of oysters, using an Illumina Golden Gate bead array. We mapped 1025 of these SNPs in one or more families, providing a nearly 10-fold increase in marker density compared to first-generation maps. We mapped 664 SNPs in two or more Pacific oyster families; all but two SNPs mapped to the same linkage group, across families, providing confidence in the assignment of markers to linkage groups. However, for genomic scaffolds that 15 have two or more mapped markers, we find that 63 of 143 such scaffolds, contain two or more SNPs that map to two or more different linkage groups. These observations suggest that the oyster genome, which has good coverage of gene-sequences, comprises a large number of miss-assembled scaffolds. A current goal is to shatter the scaffolds currently comprising the oyster genome and to re-assemble them, using information from even denser, thirdgeneration, linkage maps. EFFECTS OF BENTHIC INTERTIDAL ORGANISMS ON MARINE WATER PROPERTIES HORWITH, Micah *, and Cinde DONOHUE, WA Department of Natural Resources RUESINK, Jennifer and Alan TRIMBLE, University of Washington, Seattle, Wa 98195. Declining ocean pH has spurred research into the effects of marine carbonate chemistry on a variety of organisms, but less work has focused on the potential role of organisms in changing local carbonate chemistry. We set out to measure carbonate chemistry dynamics in marine water over each of three benthic habitats: beds of Pacific oysters (Crassostrea gigas), eelgrass (Zostera marina), and ground lacking either species. In daylight, we hypothesized that oysters would cause pH to decrease via respiratory production of CO2, and that eelgrass would cause pH to increase via photosynthetic drawdown of CO2. Over the spring and summer of 2014, we used anchored sensors, drifting sensors, and the collection of discrete water samples to characterize carbonate chemistry dynamics across the three benthic habitats in five Washington state embayments: Case Inlet, Dabob/Quilcene Bays, Port Gamble Bay, Samish Bay, and Willapa Bay. Preliminary results show substantial variation in nearshore carbonate chemistry across our five sites. These results also offer evidence that benthic organisms have important effects on local carbonate chemistry, and that these effects are stronger in shallow waters. FACT OR FICTION: PUBLIC & STAKEHOLDER PERCEPTIONS OF SHELLFISH FARMING IN THE PACIFIC NORTHWEST HUDSON, Bobbi *, Thom ALLEN, and Danna MOORE, Pacific Shellfish Institute, Olympia, WA A survey probing community views of shellfish aquaculture was sent to more than 4,000 residents in twelve coastal counties in Washington, Oregon and California. The survey explores two overarching questions: "Are these communities opposed to or supportive of continued or expanded shellfish aquaculture?" and "What are the implications for aquaculture planning and development?" The survey, conducted by Pacific Shellfish Institute and partners at Washington State University Social & Economic Sciences Research Center, elicited an unusually high 34% response rate and answers questions responding to a wide range of issues. It revealed the general view that shellfish farms "neither enhance nor detract from the scenery of coastal areas" and their "greatest benefit" is "providing locally produced seafood." This presentation 16 will highlight survey responses, with a focus on informing public outreach and education efforts related to shellfish aquaculture. A second survey has also been distributed to a broad array of local planners, federal and state natural resource agency staff, and representatives of natural resource industries, citizen groups and conservation non-government organizations. Preliminary results of this second survey will also be briefly discussed. This work is funded through Washington Sea Grant, pursuant to NOAA award #NA14OAR4170078. A COMPARISON OF BREEDING METHODS FOR THE OLYMPIA OYSTER, Ostrea lurida, USING MICROSATELLITE MARKERS JACKSON, Katherine *, Brent VADOPALAS and Steven ROBERTS, University of Washington, ALLEN, Brian, Puget Sound Restoration Fund The Olympia oyster, Ostrea lurida, is native to Puget Sound and the west coast of North America. Due to overharvesting and water quality decline, the once thriving local population has dramatically decreased and efforts to reestablish this culturally, ecologically, and economically important animal have recently commenced. While methods have been developed to breed restoration grade oysters that are genetically indistinguishable from wild populations, quantitative evaluative studies are lacking. In this study, we employed a series of population genetics tools such as expected heterozygosity, allele frequency, allele count rarefaction, and allele frequency tests using allele size data from seven microsatellite loci to investigate genetic differences among oysters reared in a restoration hatchery, a commercial hatchery, and from a wild population proximate to the targeted restoration site. . In addition, parentage analysis was used to estimate the number of successful breeders in restoration hatchery breeding groups. Results will be used to optimize Olympia oyster restoration hatchery breeding designs. ADVANCES IN ROCK SCALLOP Crassadoma gigantea CULTURE: SEED PRODUCTION AND INDUCTION OF TRIPLOIDY JACKSON, Molly*, Joth DAVIS, Brent VADOPALAS, and Sara WYCKOFF, School of Fisheries and Aquatic Sciences, University of Washington, Seattle, WA 98195 There is rapidly growing interest in aquaculture development of the Purple-hinged rock scallop (Crassadoma gigantea), a species with enormous market potential in all four U.S. states bordering the Pacific Ocean. However, culture of a native species in close proximity to wild populations can result in negative genetic interactions unless cultured individuals are effectively sterile. To investigate the use of ploidy manipulation to induce sterility, we conducted a series of trials to optimize triploid induction in rock scallops. We compared survival and ploidy from groups of rock scallops treated with eight concentrations of 6-dimethylaminopurine (6-DMAP) to block extrusion of the second polar body during meiosis. In trial 1, survival ranged between 5 17 and 40% of untreated controls, and the percentage of triploids ranged from 10-70%. We determined an optimal treatment of 425 μM 6-DMAP for 15 min, commencing at approximately 60 min post-fertilization at 17 °C. Future work will include production of diploid and triploid rock scallop seed for larger scale trials to compare growth, reproductive status, and survival at multiple growout sites, and development of tetraploid rock scallops for production of bred triploids. SHORT TERM EFFECTS OF OCEAN ACIDIFICATION ON OYSTER PREDATION BY JAPANESE OYSTER DRILLS (OCINEBRELLUS INORNATUS) KING, Derek E.*, Eric THUSENE, THE EVERGREEN STATE COLLEGE Ocean Acidification (OA) has been identified as a threat to the nutrient rich waters of Puget Sound and its host of diverse ecologically and commercially valuable bivalve populations. Limited research has been done looking at how these ocean chemistry conditions, specifically in the short term, could affect predation rates on bivalves by invasive species. This undergraduate research explores the Japanese oyster drill’s (Ocinebrellus inornatus) predatory response to short term exposures to a low PH environment while preying on juvenile pacific oysters (Cressostrea gigas). Through looking at these short term exposures (~24 hours) of a simulated OA influenced environment this study sought to identify potential future short term drill predation risks to commercial and stock rebuilding efforts of bivalves. All trial work, observations, and experiments were done in a laboratory setting using cold rooms and controlled aquariums for holding animals, treatments and controls; GoPro cameras were used to monitoring behavior and drill time; SEM Imagery was used to analyze and measure partially drilled oyster shells under different conditions. This research brings up a discussion over OA’s short term potential impact on invasive oyster drill predation, as well as highlights an area of study that may need more attention to shrink the knowledge gaps in this complex environmental problem. TALES OF A TALKING OYSTER: ENGAGING TIDELAND OWNERS TO MONITOR FECAL COLIFORM and Vibrio parahaemolyticus BACTERIA IN THEIR SHELLFISH. KING, Teri *, Washington Sea Grant, University of Washington A SIMPLE TECHNIQUE TO REDUCE OYSTER MORTALITY AND DECREASE VIBRIO LEVELS DURING THE SUMMER GROWING SEASON. HANSEN, John*, South Sound Mariculture KING, Teri, Washington Sea Grant, University of Washington 18 COMMUNITY SHELLFISH FARMS: A SPRINGBOARD FOR MULTIPLE OUTREACH OPPORTUNITIES IN WATERSHED PROTECTION DISTRICTS. KING, Derek*, Puget Sound Restoration Fund ALLEN, Brian, Puget Sound Restoration Fund We will showcase some of the many ways this community farm project has successfully engaged the public using oysters and shellfish aquaculture in the Henderson Inlet and Nisqually Watershed Protection Districts in Thurston County, WA. PLACE-BASED MARINE EDUCATION: REINFORCING STUDENT UNDERSTANDING THROUGH A LOCAL FRAMEWORK AND CONTEXTUALIZED DATA MCDONALD, P. Sean* and Alexandra DOTY, Program on the Environment, School of Aquatic and Fishery Sciences, University of Washington, Seattle, WA 98195. ARMSTRONG, Janet and David ARMSTRONG, School of Aquatic and Fishery Sciences, University of Washington, Seattle, WA 98195. LONG, Christopher, National Oceanic and Atmospheric Administration, Kodiak, AK 99615. Place-based education offers a localized framework for environmental education that can contextualize learning and engage students directly in pertinent issues. Scientists conducting research are frequently required to develop education and outreach materials as part of funding requirements. These scientific studies provide useful examples and demonstrate realworld applications of important concepts. In the present example, we were tasked with producing education and outreach materials in support of funded research on blue king crab (Paralithodes platypus). Our objective was to develop place-based lesson plans and explore how best to use data collected as part of fisheries research cruises to stimulate environmental stewardship and engagement students in local issues. To this end, qualitative interviews were conducted with high school science teachers and a meta-analysis of current literature about place-based education was performed. We developed several lesson plans about blue king crab for Alaskan high schools. Our experience supports other work indicating that place-based examples are effective tools for solidifying student understanding and facilitating student engagement in the classroom. We suggest that community involvement and local cultural knowledge are invaluable for creating impactful place-based lesson plans. Moreover, we contend that place-based lessons are most effective when created by a community in situ, and when guided but not dominated by standards and requirements. 19 NARROWING THE SEARCH FOR INVASIVE GREEN CRAB: HABITAT MAPPING WITH IMPLICATIONS FOR MONITORING EFFORTS MCDONALD, P. Sean *, School of Aquatic and Fishery Sciences, Program on the Environment, University of Washington, Seattle, WA 98195. TRAN, Chris and Jeffrey WOO, University of Washington, Seattle, WA 98195. ADAMS, Jeffrey, Washington Sea Grant, University of Washington, Seattle, WA 98195. The invasive European green crab (Carcinus maenas) was first detected in Washington waters in 1998 after warm El Niño currents spread larvae of California populations as far north as Vancouver Island. Because of perceived risks to coastal resources, the green crab was designated a deleterious species in Washington State, which imposed strict constraints on possession and transport and authorized the Washington Department of Fish and Wildlife to monitor and control the species. Most outer coast populations had limited establishment success, persisting as either small or geographically constrained populations, and no green crab were found in inland waters of Puget Sound; consequently, monitoring and control efforts were curtailed. However, in 2012, Canadian wildlife officials discovered a population of green crab in Sooke Inlet near Victoria, British Columbia, well within the Strait of Juan de Fuca. The purpose of the present study was to develop a habitat suitability map to inform monitoring and public outreach efforts. To this end, students from the University of Washington worked with staff from Washington Sea Grant and the UW School of Aquatic and Fishery Sciences to identify and prioritize suitable green crab habitat using coarse physical, biological, and access characteristics that could be observed in aerial and satellite imagery. Nearly 100 locations appear to have high likelihood of being suitable for European green crab establishment. In August 2013, select high priority sites on the Strait of Juan de Fuca and around Hood Canal were visually surveyed and found no green crab molts. Results of this work and ongoing efforts are discussed in the context of developing a new citizen science monitoring program for green crab in Puget Sound. GEODUCK AQUACULTURE IN PUGET SOUND: NEXT STEPS TOWARD INTEGRATING SOCIAL, POLICY, and ECOLOGICAL DIMENSIONS MCDONALD, P. Sean *, School of Aquatic and Fishery Sciences, Program on the Environment, University of Washington, Seattle, WA 98195. RYAN, Clare, School of Environmental and Forest Sciences, Program on the Environment, University of Washington, Seattle, WA 98195. Intertidal aquaculture of geoduck clams (Panopea generosa) poses an economically lucrative, yet scientifically, socially, and politically challenging resource management issue. Expansion of intertidal geoduck aquaculture operations in habitats of Puget Sound (Washington) in the past decade has raised concern among managers, conservation organizations, and the public 20 regarding practices that may directly or indirectly alter nearshore ecosystems. In Washington State, a legislatively mandated Geoduck Aquaculture Research (GAR) program addressed some of the ecological aspects of geoduck aquaculture, but did not explicitly consider broader policy and social dimensions of the issue. The objectives of this recently-initiated project are to: 1) conduct a policy analysis of aquaculture management approaches; 2) conduct interviews in order to identify and describe Puget Sound stakeholder interests and concerns; and 3) integrate these results with available ecological studies. The project is designed as an educational experience for students in the Environmental Management (EM) Graduate Certificate offered by the University of Washington’s Program on the Environment. An interdisciplinary team of 45 graduate students will contribute to the project in close collaboration with partner agencies and other stakeholders. Once completed, synthesis of project results with complimentary ecological research will yield a more comprehensive understanding of the socio-environmental context of geoduck aquaculture in Puget Sound. AGE ESTIMATION AND SHELL MORPHOLOGY OF THE PACIFIC GEODUCK Panopea generosa IN CALIFORNIA NAVAS, Gabriela, Moss Landing Marine Laboratories The Pacific geoduck clam, Panopea generosa, is found in intertidal to subtidal soft substrates from Alaska to Baja California. This clam is one of the largest burrowing bivalves in the world with a siphon length of up to 1m allowing it to burrow deep into the sediment. Their siphon is highly sought after as a delicacy in international markets. While there is no commercial take of P. generosa in California, a recreational fishery exists despite the paucity of data relevant to management. The objective of this study is to shed light onto California geoduck age and shell morphology variation in comparison to populations from Washington and Mexico where geoducks are a commercial fishery. Preliminary morphology results indicate a significant difference among populations in southern California sites (Bartlett’s test, Wilk’s λ= 0.0258; p < 0.001). ELEVATED TEMPERATURE AND PCO2 ALTER ATTACHMENT STRENGTH OF TWO Mytilus SPECIES NEWCOMB, L.A.*, GEORGE, M.N., O’DONNELL, M.J., and E. CARRINGTON, University of Washington, Seattle, WA 98195 Mussels form strong byssal threads to attach themselves to substrate in order to survive in their hydrodynamic environment. Weak attachment causes mussel fall-off from rocky shores and aquaculture lines, prompting our study into what environmental factors cause attachment to weaken. Our previous lab experiments showed both elevated temperature and pCO2 reduce thread strength. When acting in combination, the stronger temperature effect dominates; the 21 pCO2 signal is apparent only when temperatures are benign. Here we explore in greater detail how elevated temperature effects mussel attachment strength in two commercially farmed bay mussel species in Washington State, the native Mytilus trossulus and introduced M. galloprovincialis. Mussels were grown in a controlled laboratory setting under a range of temperatures from 10˚C and 25˚C. We found at temperatures above 22.5˚C, byssal threads of M. trossulus were 60% weaker than threads produced at 10˚C. Mytilus galloprovincialis thread strength did not depend on temperature over the range tested. Elevated temperature reduced the rate of byssal thread production in M. trossulus, but increased production in M. galloprovincialis. Future increases in ocean temperature and summer highs could challenge the attachment ability of to our native bay mussel species and favor the establishment of the introduced mussel species. TURNING THE HEADLIGHTS ON HIGH: IMPROVING OA OBSERVATION SYSTEMS IN SUPPORT OF PACIFIC COAST SHELLFISH GROWERS NEWTON, Jan, University of Washington and NANOOS Providing data on real-time ocean conditions to shellfish growers has been likened to putting headlights on a car. We are ready now to take the next step, figuratively, putting the headlights on ‘high’. The U.S. Integrated Ocean Observing System (IOOS) and NOAA’s Ocean Acidification Program (OAP) have worked together to sponsor a competition for Marine Sensor Innovation (MSI). Working with shellfish growers and the PCSGA, a successful proposal was submitted that covers developing new sensor capability in the region including Alaska, Washington, Oregon, and California. Current efforts leveraging federal IOOS and OAP dollars with state of WA and OR investments from their respective legislators have been focused on establishing a regional ocean acidification measurement “expert” in each region and to establish on-site monitoring equipment in hatcheries and shellfish farms. Under the new funding, that capacity will be sustained and enhanced, including working with a scientist and a sensor industry to develop a new, lower cost pCO2 sensor. We will: 1) Develop new lower cost and higher accuracy sensor technology for OA monitoring and expand them to new sites; 2) Utilize regional partnerships of users and local experts to implement and provide Quality Assurance/Quality Control (QA/QC) tests of the new OA sensors; 3) Establish data handling and dissemination mechanisms that provide both user-friendly and standards-based web service access that are exportable from the Pacific Coast module to the entirety of U.S. Integrated Ocean Observing System (IOOS); and 4) Provide education and outreach services to stakeholders concerned about and potentially impacted by OA. The Pacific Coast Shellfish Growers Association is a funded partner in this work and we anticipate updates and involvement as this 3-year project unfolds. 22 CHARACTERIZING GENOME-WIDE DNA METHYLATION PATTERNS TO INCREASE UNDERSTANDING OF EPIGENETIC VARIATION IN THE PACIFIC OYSTER OLSON, Claire*, ROBERTS, Steven, University of Washington, School of Aquatic and Fishery Sciences The Pacific oyster Crassostrea gigas is exposed to environmental variations including changes in temperature, pH, and oxygen levels, making this organism a key bioindicator species. Furthermore, C. gigas is an excellent model to study environmental effects on genetic modifications because genomic resources for this species are available and epigenetic modifications by DNA methylation have been described. Epigenetics, or DNA modifications that change gene expression without altering the underlying nucleotide sequence, includes DNA modifications by methyl groups and histones. DNA methylation in particular is established to change gene expression as an adaptive mechanism in response to environmental changes. Furthermore, DNA methylation patterns and functions are variable across invertebrate taxa. We have characterized the genome-wide DNA methylation profile in a single cell type (male gamete cells) and examined the relationship between DNA methylation and transcript expression. Genome-wide DNA methylation patterns of C. gigas have also been examined during oyster developmental stages (sperm and larvae), indicating the presence of lineagespecific methylation profiles. Our results provide a comprehensive characterization of the distribution of DNA methylation in a single cell type and that methylation is likely involved in gene regulatory activity and is heritable. A better understanding of epigenetic modifications in C. gigas will provide important information on molecular processes throughout the oyster development and will improve our ability to predict ecosystem responses to the aquatic environment. MANILA CLAM RECRUITMENT, SURVIVAL, GROWTH, CONDITION AND HARVEST EFFICIENCY IN WILLAPA BAY AS AFFECTED BY GRAVEL TYPE, CRUSHED SHELL AND LIME AMENDMENT, Zostera japonica CONTROL AND PREDATOR PROTECTION. Kim PATTEN*, Scott NORELIUS, Washington State University Manila clams, Ruditapes philippinarum, have become an important commercial species in Willapa Bay WA. Using a series of replicated field trials, studies were conducted to assess effects of Japanese eelgrass (Zostera japonica) removal, gravel type (round vs crushed), supplemental oyster shell (0 to 20 m3/ha), liming (0 to 3000 kg/ha), and predator protection (netting and screening) on recruitment and early survival. Few significant differences in settlement and overwinter survival were noted from any of the treatment variables. No specific variable was identified as the main driver in post-settlement mortality. Z. japonica, however, was determined to have major detrimental effects on the growth of young clams, total commercial clam harvests, clam quality and clam harvest efficiency. The significance of this effect was site and age-class dependent. The potential economic impact of a Z. japonica infestation of a Manila clam shellfish bed was ~$47,000/ha for each crop harvest cycle. 23 JAPANESE EELGRASS MANAGEMENT IN WILLAPA BAY FOR 2014: TREATMENT SUCCESS, NONTARGET IMPACTS AND ECOLOGICAL IMPLICATIONS. PATTEN, Kim*, and Scott NORELIUS, Washington State University In 2014, the Willapa Bay shellfish industry treated ~400 acres of commercial clam beds infested with Japanese eelgrass (Zostera japonica) with the herbicide imazamox. The perimeters of most of the treated beds were assessed for the percent of offsite movement of imazamox based on tracing for any visual detection of affected eelgrass outside the treated areas. Movement offsite was very limited, and only occurred along the low ends with drainage swales. Treatment rate averaged 10 oz of product per acre and efficacy across all sites was close to 100%. Two additional studies were conducted using paired plots (treated vs. untreated) on non-graveled tide flats. In one study, forage behavior of green sturgeon (n=5, plot size 0.2 to 1 ha) was assessed bimonthly based on feeding pit densities. Sites with Z. japonica removed had up to an order of magnitude greater sturgeon foraging than untreated sites. In the second study (n=3, plot size = 2 ha) comparisons were made of water flow along the sediment surface, dissolved O2, short and long-term sedimentation rates, fish and bird usage, and shellfish growth rates. Preliminary results of those findings will be presented. USING QUALITATIVE MODELS TO EVALUATE ENVIRONMENT-AQUACULTURE LINKAGES IN SOUTH PUGET SOUND AND WILLAPA BAY, WASHINGTON REUM, Jonathan C. P., Bridget FERRISS, Wei CHENG, P. Sean MCDONALD, Neil BANAS, University of Washington, Seattle, WA HARVEY, Chris, Northwest Fisheries Science Center, NOAA, Seattle, WA Identifying linkages between aquaculture and environmental variables that relate habitat alteration, climate variability, and watershed development is necessary for anticipating and mitigating against potential future change. In addition, understanding the influence of evolving aquaculture methods on coastal food webs can inform best practices, spatial marine planning efforts, and offer a fuller picture of the ecosystem context aquaculture operates within. Here, we present a qualitative modeling method that facilitates the integration of diverse data sources into networks that describe key variables and their interactions with aquaculture. The models consist of qualitative linkages (i.e., positive, negative, and neutral interactions) that may correspond to any type of variable (e.g., trophic, nontrophic, or purely conceptual). The resulting networks offer a summary of our current understanding of key system attributes, can help predict the outcome of system components to qualitative changes in perturbed variables, and help identify linkages that disproportionately influence system responses. The models permit only a limited number of key variables, and therefore demand conceptual clarity and an understanding of the causal connections between the perturbed and ecosystem components of interest. We demonstrate the method using models developed around aquaculture in Willapa Bay and South Puget Sound, and compare system responses to scenarios of future change. 24 SEASONAL VARIATION OF THE GENUS Dinophysis WITHIN PUGET SOUND, WASHINGTON: UNDERSTANDING HARMFUL ALGAL BLOOMS THROUGH SPECIES IDENTIFICATION RUNYAN, Jennifer*, THE EVERGREEN STATE COLLEGE Dinophysis is a dinoflagellate that can exude okadaic acid and dinophysistoxins causing Diarrhetic Shellfish Poisoning. Eight species of Dinophysis are found in Puget Sound houses and six out of the eight species are toxic. Seasonality of Dinophysis was examined at Quartermaster Harbor, Sequim Bay, and Penn Cove between 2012-2013. Changes in phytoplankton community composition before, during and after the presence of Dinophysis were analyzed to note any genera that may act as a precursor to the presence of Dinophysis. Lastly, species richness, evenness, and diversity were observed during Dinophysis’ presence and absence. Results showed Dinophysis spp. did vary seasonally and by site. A difference in species richness was found when Dinophysis was present at two of the three sites. Dinophysis may be predictive of certain estuarine conditions but further research is needed. A PILOT TRIAL TO ASSESS THE VERTICAL DISTRIBUTION OF PACIFIC HERRING(Clupea pallasii) SPAWN DEPOSITION IN DABOB BAY AND IMPLICATIONS FOR GEODUCK (Panopea generosa) FISHERY MANAGEMENT SIZEMORE, Bob * and Ocean EVENINGSONG, Washington Department of Fish and Wildlife, Olympia, WA 98501. In the commercial geoduck fishery, no-harvest zones have been established in nearshore subtidal areas to protect sensitive habitats such as eelgrass beds and Pacific herring spawning areas. State and tribal managers have adopted specific standard nearshore geoduck harvest depth restrictions for areas that have documented Pacific herring spawning grounds. The current restriction is no geoduck harvest in waters shallow of -25 feet year-round and -35 feet during herring spawning season (water depths corrected to mean lower low water - MLLW). These restrictions are based on historic herring spawn deposition data from rake samples that do not have precise spawning depth information. In this pilot study, scuba divers surveyed herring spawn deposition intensity in one foot vertical water depth increments from the +2 to 46 foot water depths, in Dabob Bay, WA, an area known to have consistent and abundant herring spawn deposition. The maximum water depth of medium to heavy spawn deposition was -31 feet (MLLW) and light spawn deposition was noted at a maximum depth of -42 feet (MLLW). This is the first time that scuba has been used to survey Pacific herring spawn deposition in Washington and this method may be useful to produce site specific information along commercial geoduck tracts. For example, geoduck harvest might be restricted to -42 feet (MLLW) or deeper if this pilot study site was targeted for commercial harvest. Other implications for geoduck management and stewardship of marine resources will be discussed. 25 INORGANIC ARSENIC LEVELS IN GEODUCK TISSUES: AN AGE COMPARISON STEVICK, Bethany*, Washington Department of Fish and Wildlife, Olympia, WA 98501. BORCHERT, Jerry and David MCBRIDE. Department of Health, Olympia, WA 98504. A shellfish ban on all Pacific coast shellfish imports was imposed by China in December 2013 due to concerns regarding PSP and arsenic levels in geoduck shipments. It was determined that the geoduck with high levels of inorganic arsenic that contributed to the ban originated from the Redondo commercial geoduck tract in Puget Sound, Washington. The ban was lifted in May 2014, and since the ban, NOAA has now implemented procedures to certify areas that test below China’s inorganic arsenic standard. Inorganic arsenic testing has now been conducted throughout Puget Sound by the Washington Department of Health, on both wildstock and farmed populations, and in general, arsenic accumulations are greater in wild geoduck than in farmed geoduck. Since farmed geoduck are relatively young compared to wildstock populations, age could be a contributing factor in arsenic levels in tissue. We have initiated an opportunistic study to answer the question: do older geoduck contain higher levels of inorganic arsenic? We will age shells from geoduck harvested for arsenic testing (both farmed and wildstock) from Puget Sound and Alaska, which will provide empirical data to address the question of age vs. inorganic arsenic levels. WANTED: COMPILED WEST COAST SHELLFISH RESOURCES AND FARMS ON GIS SUHRBIER, Andy, and Hannah FAULKNER, Pacific Shellfish Institute, Olympia, WA A Summary of work completed to date on compiled shellfish operations and resources on the U.S. west coast. Data include shellfish operations in California, Oregon and Washington from many data sources. The data is intended to support coastal and marine spatial planning (CMSP) activities along the West Coast. Also to assist West Coast decision-makers to understand the interrelationships of the multiple uses of ocean and coastal areas. Data has been supplied to DOE oil spill program to help protect shellfish resources in the their geographical response plan. Long-term goals of the project are to enhance ecologically and socially sustainable development of West Coast shellfish aquaculture and increase public understanding and support for shellfish related activities. 26 UNDERSTANDING INTERTIDAL HABITAT USE BY FISH AND CRAB SPECIES IN PACIFIC NORTHWEST ESTUARIES SUND, Daniel *,Marine Resource Management, College of Earth, Ocean, and Atmospheric Sciences, Oregon State University, Corvallis, OR DUMBAULD, Brett, USDA - Agricultural Research Service, Hatfield Marine Science Center, Newport, OR Zostera japonica is a non-native eelgrass brought in from Japan that is increasing in range and density in Pacific Northwest (PNW) estuaries. Eelgrass habitat provides important ecosystem services for coastal communities, commercial fisheries, and water quality. Recent decisions in Washington State to treat Z. japonica as a noxious weed revealed an emergent need to establish baseline knowledge regarding its effects on the ecology of the PNW estuaries. The goals of this project were to quantify the ecological role that Z. japonica plays by (1) comparing use of estuarine habitat types including Z. japonica, native Zostera marina and clam aquaculture via paired deployment of cameras and small fish traps and (2) examine the effect of habitat type on recruitment and survival of juvenile Dungeness crab. Review of video footage yielded a total of 11 species that utilized these habitats in Willapa Bay, Washington. Habitat was found to be a significant predictor of total catch per unit effort (CPUE) for the top four observed species (Dungeness Crab, surf perch, staghorn sculpin, and Three-spine stickleback), but not enough power to discern where this habitat effect occurred. Examination of crab recruitment across intertidal habitat types indicates that recruitment of Dungeness crab is enhanced in structured eelgrass habitat when compared to clam aquaculture beds. POTENTIAL LINKAGE BETWEEN APPARENT EARLY RECOVERY OF ENDANGERED BLACK ABALONE (Haliotis cracherodii LEACH, 1814) POPULATIONS AT SAN NICOLAS ISLAND, CALIFORNIA, AND A TWELVE-YEAR HIATUS IN LONG-TERM WARMING OF THE COASTAL OCEAN OFF CALIFORNIA. VANBLARICOM*, G.R., Washington Cooperative Fish and Wildlife Research Unit, Ecosystems Branch, US Geological Survey, and School of Aquatic and Fishery Sciences, College of the Environment, University of Washington, Seattle WA USA. Since 1985 black abalone (Haliotis cracherodii Leach, 1814) have suffered mass mortalities through most of their geographic range in California and the northern Pacific coast of Baja California, Mexico. The mortality events result from withering syndrome (WS), a disease caused by a Rickettsiales-like prokaryote, Candidatus Xenohaliotis califoriensis, unknown to science before 1985 and of unknown origin. Black abalone were listed as "endangered" in 2009, largely as a result of WS effects. Black abalone populations have been monitored quantitatively at nine long-term intertidal study sites at San Nicolas Island (SNI), California since 1981. WS first appeared at SNI in spring 1992, causing 95-99% mortality in black abalone at all nine study sites over the next decade. However, recruitment was observed at SNI in 2001, and since 2002 numbers of black abalone have increased in numbers. The Island-wide count (all study sites 27 summed) recovered to ~6% of the pre-disease (1981-1991) mean count of black abalone, and counts at south shore study sites 7 and 8 recovered to ~8% and ~15% of the pre-disease mean count, respectively, through 2013. Since ~2003 the previously significant warming pattern of the northeastern Pacific Ocean, including the entire geographic range of black abalone, has been stalled. Regional mean sea surface temperatures declined at an annual rate of 0.06-0.08°C from 2003 to at least 2011. The shift from ocean warming to the post-2003 hiatus in warming was correlated with a shift from positive phase to negative phase in the Interdecadal Pacific Oscillation (IPO). IPO is analogous to the Pacific Decadal Oscillation (PDO) of the North Pacific region, but the IPO encompasses data and processes from the entire tropical and temperate Pacific Ocean in both hemispheres. Under negative phase IPO conditions, much of the heat caused by atmospheric warming is sequestered in deep ocean waters, primarily below 300 m, with waters from the surface to ~300 m showing a cooling trend. Thus, despite the imbalance in the earth’s heat budget and the steady increase in overall global temperature and heat content, the negative IPO phase is associated with ocean circulation patterns resulting in reduced ocean surface heating rates or stasis on a global scale, and cooling on some regional scales. Previous negative IPO phases have extended for 25-30 years, with the most recent previous episode extending from 1947 to 1977. If the present negative IPO phase follows form, it is not unreasonable to anticipate persistence for as much as another decade, although such projections are entirely speculative. The ecological and oceanographic mechanisms that may link negative IPO phase conditions to abalone population growth are unknown at present. It is suggested that ocean conditions associated with the negative IPO phase may favor improved recruitment and survival rates for black abalone, and likely for other commercially significant northeastern Pacific abalone species as well. It is also suggested that shellfishery managers and marine conservation interests recognize the potential value of current ocean conditions for implementing policies and actions to recover and restore imperiled abalone populations, including the endangered black abalone. IMPROVED ANALYSIS OF CROSSBREEDING TRIALS TO INCREASE THE YIELD OF FARMED PACIFIC OYSTERS YIN, Xiaoshen * and Dennis HEDGECOCK, University of Southern California Hybrid Pacific oysters are high-yielding and tolerant of environmental stresses, implicating crossbreeding of intentionally inbred lines as an important way to improve farmed oyster production (Hedgecock & Davis 2007). Identifying elite inbred parent lines that produce highperforming hybrid seed requires diallel or factorial test crosses among lines, each acting as both a male and a female parent. The diallel allows variance in yield, among test hybrid families, to be partitioned by ANOVA into causal genetic components--principally, general (additive) combining ability, special (non-additive) combining ability, and reciprocal effects. However, ANOVA is extremely sensitive to missing information, which arises, for example, from variation in reproductive success of parent lines (missing rows or columns in the diallel matrix). To resolve this issue, we apply a general Bayesian approach, BayesDiallel (Lenarcic et al 2012), to the analysis of diallel results. A BayesDiallel analysis of yield at nearly one-year of age for 44 of 28 72 possible hybrids from an incomplete 9×9 diallel provides estimates of combining abilities for all 9 parent lines and explains 95.1% of observed yield, which suggests that BayesDiallel is robust to missing information. This analysis was not even possible with traditional ANOVA. In the analysis of a complete 5×5 diallel subset of this cross, 97.0% of observed yield is explained by BayesDiallel, compared to only 57.0% by ANOVA, indicating that BayesDiallel performs much better than ANOVA, even in analyzing a complete diallel. Therefore, BayesDiallel can estimate phenotypes of offspring more accurately, regardless of diallel completeness, which will facilitate selection of superior parent lines for producing high-yielding, hybrid, Pacific oyster seed. 29
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