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Fish egg injection as an alternative exposure route for early life stage toxicity studies: Description of two unique methods: Chapter 4

In the environment, lipophilic contaminants such as halogenated aromatic hydrocarbons (HAHs, e.g., polychlorinated biphenyls, PCBs) and polycyclic aromatic hydrocarbons (PAHs, e.g., benzo[a]pyrene) readily bioaccumulate in fish, and the bioaccumulation of these lipophilic chemicals by adult fish may have significant consequences on the development and survival of their offspring. Halogenated and polycyclic aromatic hydrocarbons translocate from adult female body stores into eggs during oocyte maturation, and early life stages of fish are often more sensitive than adults to the toxicity of these chemicals. Thus, the presence of persistent, bioaccumulative contaminants in the environment may pose a risk to fish early life stage survival and ultimately reduce recruitment into the adult population. Typically, standard early life stage toxicity studies exposed embryos, larvae, and juveniles to graded concentrations of waterborne toxicants, and dose-response relationships are based on the concentrations of chemicals in the water. However, use of waterborne exposure to assess the toxicity of persistent, bioaccumulative contaminants, such as HAHs and PAHs, has two significant drawbacks. First, uptake of hydrophobic chemicals, such as HAHs and PAHs, into the developing embryo from water is not a significant route of exposure in the environment since concentrations of these chemicals freely dissolved in water are extremely low. Rather, maternal deposition into developing oocytes is the most significant source of these chemicals to the embryo. Second, the dose received by the target tissue, in this case the developing embryo, is the most accurate predictor of the toxic response, and since extrapolation from water concentrations of the chemical to egg concentrations is required, the exact dose received by the embryo can only be estimated, often with large uncertainty. Due to these drawbacks, it is important to develop an alternative exposure method that will directly expose the developing embryo without the need to chronically expose adult fish with subsequent natural deposition of hydrophobic chemicals into the oocytes. Fish egg injection provides this exposure route. Embryos are exposed directly after fertilization with known doses of contaminants, the dose is delivered prior to critical developmental events, and extrapolation of the dose received by the embryo is not needed. We have developed two unique fish egg injection methods as alternative routes of exposure for fish early life stage toxicity studies of lipophilic environmental contaminants. With either method, individual fish eggs are injected with a known dose of chemical. The first approach, a microinjection method, originally developed to assess the developmental toxicity of HAH congeners to early life stages of salmonids, utilizes micro-syringes, 30- gauge stainless steel injection needles, and micro- to nanoliter injection volume. The second approach, a nano-injection method, utilizes glass capillary micropipettes with 2 to 10 µm tips as injection needles, and nano- to picoliter injection volume, allowing injection of nearly any size of fish egg. Both of these egg injection methods allow an investigator to assess the toxicity of lipophilic environmental contaminants to early life stages of fish in a manner that realistically reflects environmental exposure and allows accurate quantitation of the dose to the developing embryo. These injection techniques, however, are not limited to use with only lipophilic chemicals. Since the developmental toxicity of many environmental contaminants ultimately depends on the dose received by the embryo, these egg injection methods could serve as a realistic exposure route in many fish early life stage toxicity studies.

Book chapter↗

Behavioral avoidance as evidence of injury to fishery resources: Applications to natural resource damage assessment

Natural Resource Damage Assessment (NRDA) provisions enacted under Comprehensive Environmental Response Compensation and Liability Act (CERCLA) and the Oil Pollution Act (OPA) empower natural resource trustees to seek compensation for environmental injury resulting from the release of oil or hazardous substances. Under NRDA regulations promulgated under CERCLA, fish avoidance behavior is recognized as an accepted injury, and may be used to support damage claims. In support of an ongoing damage assessment, tests were conducted to determine if avoidance of ambient metals concentrations may contribute to reductions in local salmonid populations. In laboratory tests, rainbow trout ( Oncorhynchus mykiss ) and brown trout ( Salmo trutta ) avoided mixtures of metals (Cd, Cu, Pb, and Zn) at concentrations that occur in impacted river reaches at a contaminated site (Clark Fork River, MT). Avoidance of metal contamination may contribute to population reductions and preclude restoration of instream populations by prohibiting movement of fish into contaminated areas of the river from uncontaminated tributaries. Laboratory avoidance tests were performed at two testing facilities. The similar avoidance responses observed at the two laboratories demonstrated the reproducibility of avoidance measures.

Book chapter↗

Detecting Renibacterium salmoninarum in wild brown trout by use of multiple organ samples and diagnostic methods

Renibacterium salmoninarum, the causative agent of salmonid bacterial kidney disease (BKD), is endemic in many wild trout species in northerly regions. The aim of the present study was to determine the optimal R. salmoninarum sampling/testing strategy for wild brown trout (Salmo trutta L.) populations in Iceland. Fish were netted in a lake and multiple organs—kidney, spleen, gills, oesophagus and mid-gut—were sampled and subjected to five detection tests i.e. culture, polyclonal enzyme-linked immunosorbent assay (pELISA) and three different PCR tests. The results showed that each fish had encountered R. salmoninarum but there were marked differences between results obtained depending on organ and test. The bacterium was not cultured from any kidney sample while all kidney samples were positive by pELISA. At least one organ from 92.9% of the fish tested positive by PCR. The results demonstrated that the choice of tissue and diagnostic method can dramatically influence the outcome of R. salmoninarum surveys.

Bulletin of the European Association of Fish Patho↗

Non-lethal estimation of body composition of Yukon River salmon

Because of the importance of Chinook salmon to commercial and subsistence fisheries on the Yukon River, further study of the factors that may affect the success of this species and our ability to manage the fisheries is warranted. Critical to these studies is the determination of the amount of lipids (fat) stored and available to the fish as its primary energy source for migration and spawning. Recent developments of Bioelectrical Impedance Analysis (BIA) promise a simple, non-lethal means of estimating proximate composition (e.g. fat, protein, water content) for field applications with fish. The goal of the project was to develop BIA models for Chinook salmon from the Yukon River watershed that would permit the non-lethal estimation of body proximate composition for use in field studies. Our results clearly demonstrated that BIA can be used to estimate proximate composition and energy density of salmon. While some minor refinements were suggested, the methodology can be used in a wide variety of field applications. For instance, application of the BIA models to predict energy levels of fish during their migration will allow evaluation of management programs, while also yielding data that can be used to evaluate energy use along the migratory path. Correlations of energy level with ongoing tagging, radio-tracking, and genetic studies also have the potential to allow managers and scientists to understand the relationship between fat content and distance to spawning location. These models have the potential for application to this species in other river systems. They also provide tools for a variety of other scientific investigation such as: 1) differences in energy stores in spawning and recruitment success; 2) effects of global warming on migratory salmonid stocks; and 3) differences in annual flow and temperature regiments upon migratory energy costs and resulting recruitment success.

Alaska, Yukon↗

Food web conceptual model

This chapter describes a general model of food webs within tidal wetlands and represents how physical features of the wetland affect the structure and function of the food web. This conceptual model focuses on how the food web provides support for (or may reduce support for) threatened fish species. This model is part of a suite of conceptual models designed to guide monitoring of restoration sites throughout the San Francisco Estuary (SFE), but particularly within the Sacramento-San Joaquin Delta (Delta) and Suisun Marsh. The conceptual models have been developed based on the Delta Regional Ecosystem Restoration Implementation Plan (DRERIP) models, and are designed to aid in the identification and evaluation of monitoring metrics for tidal wetland restoration projects. Many tidal restoration sites in the Delta are being constructed to comply with environmental regulatory requirements associated with the operation of the Central Valley Project and State Water Project. These include the Biological Opinions for Delta Smelt (Hypomesus transpacificus) and salmonids (U.S. Fish and Wildlife Service 2008; National Marine Fisheries Service 2009), and the Incidental Take Permit for Longfin Smelt (Spirinchus thaleichthyes) (California Department of Fish and Wildlife 2009). These regulatory requirements are based on the hypothesis that the decline of listed fish species is due in part to a decline in productivity of the food web (phytoplankton and zooplankton in particular) or alterations in the food web such that production is consumed by other species in the Estuary (Sommer et al. 2007; Baxter et al. 2010; Brown et al. 2016a). Intertidal wetlands and shallow subtidal habitat can be highly productive, so restoring areas of tidal wetlands may result in a net increase in productivity that will provide food web support for these fish species. However, other factors such as invasive bivalves that reduce phytoplankton and zooplankton biomass and invasive predatory fishes that may compete with or prey upon listed fishes can limit the utility of tidal wetlands for food web support (Lucas and Thompson 2012; Herbold et al. 2014). This model utilizes information from the previous DRERIP models for Delta food webs (Durand 2008) and tidal wetlands (Kneib et al. 2008), an updated DRERIP model (Durand 2015), and the State of BayDelta Science 2016 review of recent Delta food web literature (Brown et al. 2016a).

Interagency Ecological Program Technical Report↗

Estimation of stream conditions in tributaries of the Klamath River, northern California

Because of their critical ecological role, stream temperature and discharge are requisite inputs for models of salmonid population dynamics. Coho Salmon inhabiting the Klamath Basin spend much of their freshwater life cycle inhabiting tributaries, but environmental data are often absent or only seasonally available at these locations. To address this information gap, we constructed daily averaged water temperature models that used simulated meteorological data to estimate daily tributary temperatures, and we used flow differentials recorded on the mainstem Klamath River to estimate daily tributary discharge. Observed temperature data were available for fourteen of the major salmon bearing tributaries, which enabled estimation of tributary-specific model parameters at those locations. Water temperature data from six mid-Klamath Basin tributaries were used to estimate a global set of parameters for predicting water temperatures in the remaining tributaries. The resulting parameter sets were used to simulate water temperatures for each of 75 tributaries from 1980-2015. Goodness-of-fit statistics computed from a cross-validation analysis demonstrated a high precision of the tributary-specific models in predicting temperature in unobserved years and of the global model in predicting temperatures in unobserved streams. Klamath River discharge has been monitored by four gages that broadly intersperse the 292 kilometers from the Iron Gate Dam to the Klamath River mouth. These gages defined the upstream and downstream margins of three reaches. Daily discharge of tributaries within a reach was estimated from 1980-2015 based on drainage-area proportionate allocations of the discharge differential between the upstream and downstream margin. Comparisons with measured discharge on Indian Creek, a moderate-sized tributary with naturally regulated flows, revealed that the estimates effectively approximated both the variability and magnitude of discharge.

California↗

Natural resource condition assessment: Olympic National Park

The Natural Resource Assessment Program aims to document condition and trends of selected park resources while identifying emerging issues and information needs. This information is intended to serve as a platform for natural resource managers to use in developing future resource stewardship priorities and planning. Olympic National Park (OLYM) on Washington’s Olympic Peninsula protects remarkable examples of several Pacific Northwestern ecosystems, including the glacier-capped Olympic Mountains, oldgrowth temperate rainforests, pristine river systems, and wild Pacific coastline and islands. The park provides habitat for numerous plants and animals, including at least 16 animal and eight plant taxa endemic to the Olympic Peninsula. The park’s lakes and rivers support over 70 stocks of Pacific salmonids and 29 native freshwater fish species. The rocky intertidal community is one of the most complex and diverse shorelines in the United States. Although we conducted in-depth assessments on a limited number of resources, the general condition of several other physical and biological components of OLYM ecosystems is described in Chapter 2, along with an overview of park history and ecology. In Chapter 3, we describe how we chose our focal resources and measures, as well as our protocol for conducting the assessment. In Chapter 4, we provide a detailed assessment of each resource, with a summary of condition and trends. In Chapter 5, we further interpret and discuss the implications of each focal resource status, highlighting future areas for monitoring and research. We summarize the general findings from our assessments below.

Washington↗

Wind River Subbasin Restoration, Annual report of U.S. Geological Survey activities, January 2016 through December 2016

We used Passive Integrated Transponder (PIT)-tagging and a series of instream PIT-tag interrogation systems (PTISs) to investigate life-histories, populations, and efficacy of habitat restoration actions for steelhead Oncorhynchus mykiss in the Wind River subbasin, WA. Our tagging focused on parr in headwaters areas of the subbasin and our instream readers provided information on movement of these parr and other life-stages of tagged steelhead. The Wind River subbasin in southwest Washington State provides habitat for a population of wild Lower Columbia River steelhead and is an excellent watershed for long-term studies of population dynamics and responses to restoration of this wild population. No hatchery steelhead have been planted in the Wind River subbasin since 1994, and hatchery adults are estimated to be less than one percent of adults in any year (pers comm. Thomas Buehrens, Washington Department of Fish and Wildlife). Numerous restoration actions have been implemented in the subbasin, including the removal of Hemlock Dam on Trout Creek in 2009. Data from our study, and companion work by Washington Department of Fish and Wildlife (WDFW), will contribute to Bonneville Power Administration’s (BPA) Research Monitoring and Evaluation (RM&E) Program Strategy of Fish Population Status Monitoring (www.cbfish.org/ProgramStrategy.mvc/ViewProgramStrategySummary/1), specifically the substrategies of: 1) Assessing the Status and Trends of Diversity of Natural Origin Fish Populations and to Uncertainties Research regarding differing life histories of a wild steelhead population, 2) Assessing the Status and Trend of Adult Natural Origin Fish Populations, and 3) Monitoring and Evaluating the Effectiveness of Tributary Habitat Actions Relative to Environmental, Physical, or Biological Performance Objectives. During summer 2016, we sampled and PIT-tagged age-0 and age-1 steelhead parr in headwater areas of the Wind River subbasin to characterize population traits and investigate variable life-histories, including growth and parr movement downstream prior to smolting. Repeat sampling and smolt traps provide opportunities for recapture, and instream PTISs and Columbia River infrastructure provide opportunity for detection of PIT-tagged fish. Throughout the year, we maintained a series of instream PTISs to monitor movement of tagged steelhead parr, smolts, and adults. During 2016, we repaired or replaced much of our instream PTIS infrastructure that had been damaged or destroyed during a large flood event in December 2015. This included moving our upper Wind River detection site (WRU) about a kilometer downstream to a location we hope to be less susceptible to damage in high flows and that will allow grid power connection for more reliable winter operations. Detections at the instream PTISs showed trends of parr emigration during summer and fall, in addition to the expected movement of parr and smolts in spring. These data are increasing our understanding of varied life histories of juvenile steelhead; paired with other steelhead population work in the subbasin we hope to begin to understand some of the factors which may influence parr movements. Long-term monitoring of PIT-tagged fish over multiple years is providing information on contribution of various life-history strategies to smolt production and adult returns, as well as helping to identify factors influencing parr movement. Movements of PIT-tagged adult steelhead were also tracked with our instream PTISs. These data have provided information on timing of adult movements to various parts of the watershed, which is allowing us to assess adult returns to tributary watersheds within the Wind River subbasin. Determination of adult use of tributary watersheds is providing data to contribute to evaluation of the efficacy of the removal of Hemlock Dam on Trout Creek. Hemlock Dam, located at rkm 2.0 of Trout Creek was removed in summer 2009 and had contributed to hydrologic impairment of Trout Creek Evaluating restoration efforts is of interest to many managers and agencies so that funding and time are allocated for best results. The evaluation of various life-histories of Lower Columbia River steelhead within the Wind River subbasin will provide information to better track populations, and to direct habitat restoration and water allocation planning. Increasingly detailed Viable Salmonid Population information, such as that provided by PIT-tagging and instream PTISs networks like those we are building and operating in the Wind River subbasin, will provide data to inform policy and management, as life-history strategies and production bottlenecks are identified and understood.

Report↗

Smallmouth bass predation on subyearling fall Chinook salmon in Lower Granite Reservoir, 2016–2017

Predation by nonnative fishes is one factor that has been implicated in the decline of juvenile salmonids in the Pacific Northwest. Impoundment of much of the Snake and Columbia Rivers has altered food webs and created habitat favorable for species such as smallmouth bass Micropterus dolomieu. Smallmouth bass are common throughout the Columbia River basin and have become the most abundant predator in lower Snake River reservoirs (Zimmerman and Parker 1995). This is a concern for Snake River fall Chinook salmon Oncorhynchus tshawytscha (hereafter, subyearlings) that may be particularly vulnerable due to their relatively small size and because their main-stem rearing habitats often overlap or are in close proximity to habitats used by smallmouth bass (Curet 1993; Tabor et al. 1993). Concern over juvenile salmon predation spawned a number of large-scale studies to quantify its effect in the late 1980s, 1990s, and early 2000s (Poe et al. 1991; Rieman et al. 1991; Vigg et al. 1991; Fritts and Pearsons 2004; Naughton et al. 2004). Smallmouth bass predation represented 9% of total salmon consumption by predatory fishes in John Day Reservoir, Columbia River, from 1983 through 1986 (Rieman et al. 1991). In transitional habitat between the Hanford Reach of the Columbia River and McNary Reservoir, juvenile salmon (presumably subyearlings) were found in 65% of smallmouth bass (>200 mm) stomachs and comprised 59% of the diet by weight (Tabor et al. 1993). Within Lower Granite Reservoir on the Snake River, Naughton et al. (2004) showed that monthly consumption (based on weight) ranged from 5% in the upper reaches of the reservoir to 11% in the forebay. However, studies in the Snake River were conducted soon after Endangered Species Act (ESA) listing of Snake River fall Chinook salmon (NMFS 1992). During this time, fall Chinook salmon abundance was at an historic low, which may explain why consumption rates were relatively low compared to those from studies conducted in the Columbia and Yakima Rivers where abundance was higher (e.g., Tabor et al. 1993; Fritts and Pearsons 2004). We speculate that predation on subyearlings by smallmouth bass in the Snake River may have increased in recent years for several reasons. Since their ESA listing, recovery measures implemented for Snake River fall Chinook salmon have resulted in a large increase in the juvenile population (Connor et al. 2013). Considering that subyearlings probably now make up a larger portion of the forage fish population, it is plausible they should make up a larger portion of smallmouth bass diets. Second, migrating subyearlings delay downstream movement in the transition zones of the Clearwater River and Snake River for varying lengths of time (Tiffan et al. 2010), which increases their exposure and vulnerability to predators. Spatial overlap in locations of smallmouth bass and subyearlings that died during migration provides support for this (Tiffan et al. 2010). Finally, the later outmigration of subyearlings from the Clearwater River results in their presence in Lower Granite Reservoir during the warmest summer months when predation rates of smallmouth bass should be highest. In 2016 and 2017, we focused our smallmouth bass predation efforts in Lower Granite Reservoir downstream of the transition zones and the confluence area where we worked during 2012–2015. This report primarily covers results from 2017 but some results from 2016 are also included for comparison. Similar to past years, our first objective was to quantify smallmouth bass consumption rates of subyearlings, determine bass abundance, and describe bass diets. In addition, Tiffan et al. (2016a) posited that predation risk to subyearlings may be higher in shoreline habitats that are more suitable for smallmouth bass and lower in shoreline habitats that are more suitable for subyearlings. To test this hypothesis, our second objective was to examine the relationship between smallmouth bass predation and subyearling habitat suitability. Our final objective was to combine estimates of consumption with smallmouth bass abundance to derive estimates of total Chinook salmon losses to smallmouth bass for 2016 and 2017.

Idaho, Washington↗

Efficacy of injectable tulathromycin for reduction of vertical transmission of Renibacterium salmoninarum in Spring Chinook Salmon Oncorhynchus tshawytscha

Bacterial kidney disease (BKD) caused by Renibacterium salmoninarum (Rs) occurs nearly worldwide where wild or cultured salmonid fishes are present. Control of BKD is confounded by its two modes of transmission, horizontal (fish-to-fish) and vertical (from female parent to progeny via the eggs). A highly successful BKD control strategy employed in Pacific Northwest hatcheries culturing spring Chinook salmon ( Oncorhynchus tshawytscha ) includes: (1) injecting pre-spawning adults with a macrolide antibiotic to improve survival and reduce Rs infection levels, (2) broodstock culling of highly infected females and (3) improved fish husbandry. However, the future availability of the injectable macrolide antibiotic (erythromycin) used for adults is uncertain. This drug shortage has resulted in an urgent need to identify a replacement injectable antibiotic to ensure continued successful control of BKD. The research conducted was intended to provide information for addressing this need via preliminary tests of the safety and efficacy of a new macrolide antibiotic, injectable tulathromycin, which is sold under the trade name DRAXXIN® (Zoetis Animal Health). A long-term goal is to reduce or eliminate the use of antibiotic treatment in spring Chinook salmon hatchery culture. Non-treated females were included in the study to provide empirical data in support of this goal. A subset of pre-spawning spring Chinook salmon at Leavenworth NFH was injected on July 10, 2014 with DRAXXIN at 5 mg per kg body weight (31 fish, left pelvic fin clip). Another subset of females (30 fish, right pelvic fin clip) was left uninjected. The surviving fish (31 DRAXXIN-injected fish and 28 uninjected fish) were spawned between August 18 and September 2, 2014. Although there were apparent trends toward higher pre-spawn survival and lower Rs prevalence and levels for the DRAXXIN-injected females in comparison to the uninjected females, the differences were not statistically significant for any of the Rs assays used (P > 0.05). Based on USFWS enzyme-linked immnosorbent assay (ELISA) test results of kidney tissue samples from the spawning females, egg lots from DRAXXIN-injected and uninjected females were assigned to Rs vertical transmission risk groups (low, medium or high). A subset of 220 eyed eggs from each female was transferred to the Western Fisheries Research Center (USGS) on October 8, 2014, hatched and reared until the study was terminated on September 22, 2015. The study results provided no evidence that DRAXXIN injection of adult female Chinook salmon affected their fecundity, egg eye-up, or survival and growth of progeny fry. There was little evidence of Rs infection in progeny of either DRAXXIN-injected or uninjected females, so the effect of DRAXXIN injection on vertical transmission of Rs could not be assessed. To adequately evaluate the efficacy of DRAXXIN injection for reducing Rs vertical transmission to progeny, additional studies should be conducted with larger numbers of DRAXXIN-injected and uninjected Chinook salmon females with a greater range of Rs levels.

Report↗

Wind River subbasin restoration annual report of USGS activities January 2017 through December 2017

We used Passive Integrated Transponder (PIT)-tagging and a series of instream PIT-tag interrogation systems (PTISs) to investigate life-histories, populations, and efficacy of habitat restoration actions for wild Steelhead Oncorhynchus mykiss in the Wind River subbasin, WA. No hatchery Steelhead have been planted in the Wind River subbasin since 1997, and hatchery adults are estimated to be less than one percent of adults in most years (pers comm. Thomas Buehrens, Washington Department of Fish and Wildlife). Numerous restoration actions have been implemented in the subbasin, including Hemlock Dam removal on Trout Creek in 2009. Data from our study, and companion work by Washington Department of Fish and Wildlife (WDFW), are contributing to Bonneville Power Administration’s (BPA) Research Monitoring and Evaluation (RM&E) Program Strategy of Fish Population Status Monitoring (www.cbfish.org/ProgramStrategy.mvc/ViewProgramStrategySummary/1), specifically the substrategies of: 1) Assessing the Status and Trends of Diversity of Natural Origin Fish Populations and to Uncertainties Research regarding differing life histories of a wild Steelhead population, 2) Assessing the Status and Trend of Adult Natural Origin Fish Populations, and 3) Monitoring and Evaluating the Effectiveness of Tributary Habitat Actions Relative to Environmental, Physical, or Biological Performance Objectives. Our headwaters parr PIT tagging, WDFW parr, smolt, and adult tagging and our instream PTISs are providing data movements and life histories of parr, smolt, and adult Steelhead. During summer 2017, we PIT-tagged age-0 and age-1 Steelhead parr in headwater areas of the Wind River subbasin to characterize population traits and investigate life-history diversity, including growth and pre-smolt downstream movement. Repeat sampling and smolt traps provide opportunities for recapture, and instream PTISs and Columbia River infrastructure provide opportunity for detection of PIT-tagged fish. Throughout the year, we maintained a series of instream PTISs to monitor movement of tagged Steelhead parr, smolts, and adults. This included adding the second array to our upper Wind River PITS, increasing solar capacity and adding improved power cables to some sites. Detections at the instream PTISs have demonstrated trends of age-0 and age-1 parr emigration from natal areas during summer and fall, in addition to the expected movement of parr and smolts in spring. These data are increasing our understanding of varied life histories of juvenile Steelhead; paired with other Steelhead population work in the subbasin we hope to begin to understand factors which may influence parr movements. Long-term monitoring of PIT-tagged fish over multiple years is providing information on contribution of various life-history strategies to smolt production and adult returns. Movements of PIT-tagged adult Steelhead were also recorded at instream PTISs. These data have allowed us to assess adult returns to tributary watersheds within the Wind River subbasin. Determination of adult use of tributary watersheds is providing data to contribute to evaluation of the efficacy of the removal of Hemlock Dam on Trout Creek. Hemlock Dam, located at rkm 2.0 of Trout Creek was removed in summer 2009. The dam had had contributed to hydrologic impairment of Trout Creek and had potential negative effects on Steelhead. The improved upper Wind River PTIS (better site characteristics and grid power) will allow estimates of subbasin adult escapement upstream of that site. Evaluating and planning restoration efforts are of interest to many managers and agencies to ensure efficient use of resources. The evaluation of various life-histories of Lower Columbia River Steelhead within the Wind River subbasin will provide information to better track populations, and to direct habitat restoration and water allocation planning. Increasingly detailed Viable Salmonid Population information, such as that provided by PIT-tagging and instream PTISs networks like those we operate in the Wind River subbasin, will provide data to inform policy and management, as life-history strategies and production bottlenecks are identified and understood.

Washington↗

Sperm cryopreservation in fish and shellfish

Initial success in sperm cryopreservation came at about the same time for aquatic species and livestock. However, in the 50-plus years since then cryopreserved sperm of livestock has grown into a billion-dollar global industry, while despite work in some 200 species with well over 200 published reports, cryopreservation of aquatic species sperm remains essentially a research activity with little commercial application. Most research has focused on large-bodied culture and sport fishes, such as salmonids, carps, and catfishes, and mollusks such as commercially important oyster and abalone species. However, only a handful of studies have addressed sperm cryopreservation in small fishes, such as zebrafish, and in endangered species. Overall, this work has yielded techniques that are being applied with varying levels of success around the world. Barriers to expanded application include a diverse and widely distributed literature base, technical problems, small sperm volumes, variable results, a general lack of access to the technology, and most importantly, the lack of standardization in practices and reporting. The benefits of cryopreservation include at least five levels of improvements for existing industries and for creation of new industries. First, cryopreservation can be used to improve existing hatchery operations by providing sperm on demand and simplifying the timing of induced spawning. Second, frozen sperm can enhance efficient use of facilities and create new opportunities in the hatchery by eliminating the need to maintain live males, potentially freeing resources for use with females and larvae. Third, valuable genetic lineages such as endangered species, research models, or improved farmed strains can be protected by storage of frozen sperm. Fourth, cryopreservation opens the door for rapid genetic improvement. Frozen sperm can be used in breeding programs to create improved lines and shape the genetic resources available for aquaculture. Finally, cryopreserved sperm of aquatic species will at some point become an entirely new industry itself. A successful industry will require integrated practices for sample collection, refrigerated storage, freezing, thawing, rules for use and disposal, transfer agreements, and database development. Indeed the development of this new industry is currently constrained by factors including the technical requirements for scaling-up to commercial operations during the transition from research, and the absence of uniform quality control practices, industry standards, marketing and price structures, and appropriate biosecurity safeguards.

Society for Reproduction and Fertility Supplement↗

Estimating annual Ceratonova shasta mortality rates in juvenile Scott and Shasta River coho salmon that enter the Klamath River mainstem

The impacts of Ceratonova shasta on Klamath River salmonids have been the focus of tremendous research and monitoring over the past decade. Knowledge gained from the various studies has resulted in a growing suite of decision support tools that can be used to predict the prevalence of infection and mortality that can be expected from varying exposure concentrations to C. shasta waterborne spores, exposure durations, and water temperatures. More recent efforts have shown that when coupled with estimates of abundance and temporal migration timing, these tools can be used to predict population-level rates of annual mortality due to ceratomyxosis. In this report, we apply those tools to estimate annual mortality rates of juvenile Coho Salmon Oncorhynchus kisutch originating from the Scott and Shasta rivers that result from exposure to C. shasta in the mainstem Klamath River. Additionally, we simulate how a water management flow regime targeted to reduce disease risk may translate into differing estimates of mortality rates. In general, we found large variation among annual mortality rate estimates, ranging from 0% to 68% across both rivers and across all years considered in this investigation. Further, we estimate that fish from the Shasta River experienced higher mortality rates owning to their prolonged exposure history. Finally, our simulations suggest that the flow regime targeting reduced disease risk could be effective at reducing annual Coho Salmon mortality rate estimates, with the extent of the estimated reductions related to the timing of the onset of measurable in-river waterborne spore concentrations and the overlap with the Klamath River residence for each source population.

California, Oregon↗

Lake Ontario deepwater sculpin recovery: An unexpected outcome of ecosystem change

Fish population recoveries can result from ecosystem change in the absence of targeted restoration actions. In Lake Ontario, native Deepwater Sculpin Myoxecephalus thompsonii, were common in the late-1800s, but by the mid-1900s the species was possibly extirpated. During this period mineral nutrient inputs increased and piscivore abundance declined, which increased the abundance of the nonnative planktivores Alewife Alosa pseudoharengus and Rainbow Smelt Osmerus mordax. Deepwater Sculpin larvae are pelagic and vulnerable to predation by planktivores. Annual bottom trawl surveys did not capture Deepwater Sculpin from 1978 – 1995 (n = 6,666 tows) despite sampling appropriate habitat (trawl depths: 7 – 170 m). The absence of observations during this time resulted in an elevated conservation status for the species, but no restoration actions were initiated. In 1996, three individuals were caught in bottom trawls, the first observed since 1972. Since then, their abundance has increased and in 2017 they were the second most abundant Lake Ontario prey fish. The food-web changes that occurred from 1970 through the 1990s contributed to this recovery. Alewife and Rainbow Smelt abundance declined during this period due to predation by stocked salmonids and legislation that reduced nutrient inputs and food web productivity. In the 1990s, the proliferation of nonnative, filter-feeding dreissenid mussels dramatically increased water clarity. As light penetration increased, the early-spring depth distribution of Alewife and Rainbow Smelt shifted deeper, away from larval Deepwater Sculpin habitat. The intentional and unintentional changes that occurred in Lake Ontario were not targeted at Deepwater Sculpin restoration but resulted in conditions that favored the species’ recovery. While standard surveys documented the recovery, more diverse information (e.g. observations in deep habitats and early-life stages) would have improved our understanding of why the species recovered when it did. Annual Lake Ontario trawl surveys have collaboratively expanded their spatial extent and diversified habitat sampled, based on lessons learned from the Deepwater Sculpin recovery.

Lake Ontario↗

Wind River subbasin restoration: Annual report of US..Geological Survey activities, January 2018 through December 2018

We sampled juvenile wild Steelhead Oncorhynchus mykiss in headwater streams of the Wind River, WA, to characterize populations and investigate life-history metrics, particularly migratory patterns. We used Passive Integrated Transponder (PIT)-tagging and a series of instream PIT-tag interrogation systems (PTISs) to track juveniles. The Wind River subbasin is considered a wild Steelhead refuge by Washington Department of Fish and Wildlife (WDFW). No hatchery Steelhead have been planted in the Wind River subbasin since 1997, and hatchery adults are estimated to be less than one percent of spawners in most years (pers comm. Thomas Buehrens, Washington Department of Fish and Wildlife). Our repeated headwater sampling of consistent sites in the Wind River subbasin has also allowed us to track relative abundance of Brook Trout, a non-native species to the Wind River. Our work is contributing to understanding of Steelhead population response to numerous restoration actions in the subbasin, including removal of Hemlock Dam from Trout Creek in 2009, where our PTISs are helping to quantify adult response. Data from our study, and companion work by Washington Department of Fish and Wildlife, are contributing to Bonneville Power Administration’s (BPA) Research Monitoring and Evaluation (RM&E) Program Strategy of Fish Population Status Monitoring (www.cbfish.org/ProgramStrategy.mvc/ViewProgramStrategySummary/1). Specifically this work addresses the sub-strategies of: 1) Assessing the Status and Trends of Diversity of Natural Origin Fish Populations and to Uncertainties Research regarding differing life histories of a wild Steelhead population, 2) Assessing the Status and Trend of Adult Natural Origin Fish Populations, and 3) Monitoring and Evaluating the Effectiveness of Tributary Habitat Actions Relative to Environmental, Physical, or Biological Performance Objectives. Our headwaters parr PIT tagging, WDFW parr, smolt, and adult tagging and our instream PTISs are providing data on movements and life histories of parr, smolt, and adult Steelhead. During summer 2018, we PIT-tagged 1,592 age-0 and age-1 Steelhead parr in headwater areas of the Wind River subbasin to characterize population traits and investigate life-history diversity, including growth and pre-smolt downstream movement. Repeat headwater sampling and smolt trap operations provide opportunities for recapture, and instream PTISs and Columbia River infrastructure provide opportunity for detection of PIT-tagged fish. Throughout the year, we maintained a series of six instream PTISs to monitor movement of tagged Steelhead parr, smolts, and adults. Detections at the instream PTISs have demonstrated trends of age-0 and age-1 parr emigration from natal areas during summer and fall, in addition to the expected movement of parr and smolts in spring. Substantial numbers of parr make downstream movements as age-1 fish. We have estimated that from 15 to 33 percent of parr tagged as age-0 fish make downstream migrations at age-1 for additional rearing. We have estimated that from 1 to 27 percent of parr tagged as age-1 fish make downstream migrations during fall. These findings raise many questions about parr rearing strategies, habitat use, and success of these migrants and suggest a need for broader monitoring of juvenile Steelhead in some river systems to fully document juvenile production. Long-term monitoring of PIT-tagged fish is providing information on contribution of various life-history strategies to smolt production and adult returns. Movements of PIT-tagged adult Steelhead were recorded at instream PTISs. These data have allowed assessment of adult returns to tributary watersheds within the Wind River subbasin. Detection efficiency of adult PIT-tagged Steelhead at our primary adult-monitoring PTIS in Trout Creek has been greater than 92 percent during 6 of the past 7 years. This is providing excellent data to estimate adult returns to this watershed. Determination of adult use of tributary watersheds is providing data to help evaluate the efficacy of the removal of Hemlock Dam on Trout Creek. Hemlock Dam, located at rkm 2.0 of Trout Creek, was removed in summer 2009. The dam contributed to hydrologic impairment of Trout Creek and had potential negative effects on Steelhead. The improvements made to the upper Wind River PTIS (site code WRU at rkm 28.3; better site characteristics and grid power) during 2016 and 2017, and a planned new site in the Mine Reach of the upper Wind River, will allow estimates of subbasin adult escapement like those in Trout Creek. During 2018, we also completed planning and permitting with U.S. Forest Service for a new PTIS site at rkm 36 of the Wind River (the Mine Reach, mentioned above). This site will replace two sites (one in Paradise Creek and one at rkm 41 of the Wind River), which had operational challenges due to lack of adequate solar power and winter difficulties. The new Mine Reach PTIS site at rkm 36, will have better solar exposure, fewer winter operations difficulties, and provide opportunity to detect fish from juvenile sampling sites that were downstream of the previous two PTISs. The more consistent operation of the new Mine Reach PTIS site will increase our ability to estimate migrant abundance as all the juveniles tagged upstream of it will be subject to the same potential detection history, instead of three different potential detection histories as before. Additionally, with the new Mine Reach PTIS site lower in the watershed, it will subject more PIT-tagged adult Steelhead to detection and provide ability to generate a nonbiased adult-detection efficiency estimate for the WRU PTIS at rkm 28.3 of the Wind River. This will provide the opportunity to estimate yearly adult Steelhead abundance to the upper Wind watershed area. Permitting is complete and some supplies have been purchased to build and install this new site in 2019. Repeat sampling at consistent locations in the subbasin has allowed investigation into juvenile Steelhead growth patterns. Growth rates (relative change in weight) of age-0 PIT-tagged parr during summer are similar across the subbasin, but lower for age-1 parr in the Trout Creek watershed than the upper Wind River watershed. Yearly growth for parr tagged at age-0 is similar across the subbasin. Yearly growth for parr tagged at age-1 is lowest in Martha Creek, but similar elsewhere. Non-native Brook Trout are present in portions of the subbasin, chiefly the Trout Creek watershed, and repeat sampling has allowed us to index their prevalence. Percentage of catch that is Brook Trout at each of four sample sites in Trout Creek have declined from the period 1998 – 2003 to the period 2011 – 2018. There was a pattern of decline in percent of catch and number of Brook Trout at the Trout Creek sites from 2011 through 2016, though a slight upward trend during 2017 and 2018 has been evident. Evaluating and planning restoration efforts are of interest to many managers and agencies to ensure efficient use of resources. The evaluation of various life-histories of Steelhead within the Wind River subbasin will provide information to better track populations, and to direct habitat restoration and water allocation planning. Movement of Steelhead parr raises many questions regarding estimating juvenile abundance, origin, and habitat use within watersheds. Improved PTISs and focused PIT tagging of age-0 and age-1 Steelhead parr are increasingly allowing us to investigate such questions. Increasingly detailed Viable Salmonid Population information, such as that provided by PIT-tagging and instream PTISs networks like those in the Wind River subbasin, provide data to inform policy and management, as life-history strategies and production bottlenecks are identified and understood.

Washington↗

Status of Cisco in Lake Superior in 2017

This report describes the status of fish species and their habitat in Lake Superior during the reporting period of 2012-2016 in response to achievement of fish community objectives (FCOs) established by fishery managers for the lake. The overarching goal for the FCOs continued to be met as the fish community remained diverse, self-regulating, dominated by indigenous species, and able to support sustainable fisheries, although further rehabilitation of certain fish is required. The Lake Superior Lakewide Action and Management Plan classified all habitat indicators for Lake Superior as good. Primary production and zooplankton abundance were stable during the reporting period and unchanged from the two previous reporting periods, indicating the lower food web is healthy. Abundance of the invertebrates Mysis diluviana and Diporeia spp. were stable during the reporting period, and Diporeia spp. density exceeded target levels defined in the Great Lakes Water Quality Agreement. Lake Whitefish (Coregonus clupeaformis) abundance was lower than during the previous reporting period but was within the FCO target. Abundance of lean, siscowet, and humper forms of Lake Trout (Salvelinus namaycush) remained stable at levels seen in previous reporting periods. The FCO for non-indigenous salmonids was met as Chinook Salmon (Oncorhynchus tshawytscha), Coho Salmon (O. kisutch), and steelhead/Rainbow Trout (O. mykiss) were being sustained by natural reproduction, and their abundance remained stable or increased from previous reporting periods. The FCO for Walleye (Sander vitreus) was not met, although populations showed signs of improvement since the previous reporting period. The fish community in littoral areas and embayments continued to be diverse and composed mostly of indigenous species. No new invasive species were found in Lake Superior during the reporting period. Degraded embayment and tributary habitats continued to prevent achievement of the FCOs for Brook Trout (S. fontinalis) and Lake Sturgeon (Acipenser fulvescens). Barriers created for hydroelectric generation either blocked Lake Sturgeon from historically important spawning grounds or reduced stream flows necessary for its reproduction. In tributaries without man-made barriers, Lake Sturgeon was relatively abundant and appeared healthy. Attainment of the FCOs for Brook Trout and Lake Sturgeon will be challenging and can only be attained through development of large-scale management actions like those implemented for Lake Trout rehabilitation and Sea Lamprey (Petromyzon marinus) control. Sea Lamprey-control expenditures doubled in 2016 from past levels and should result in suppression of the population closer to its FCO after 2016. The prey-fish FCO appears to have been met, but biomass of nearly all prey-fish species declined from that reported for the previous reporting period and has been on a downward trajectory since 2000. Predation by Lake Trout is probably to blame for the declining biomass of prey fish. Poor recruitment by Cisco (C. artedi) over the last 15 years is exacerbating the declines in prey-fish biomass because Lake Trout must compensate for the loss of Cisco by consuming other, less-abundant prey fish.

Lake Superior↗

State of Lake Superior ecosystem in 2017

This report describes the status of fish species and their habitat in Lake Superior during the reporting period of 2012-2016 in response to achievement of fish community objectives (FCOs) established by fishery managers for the lake. The overarching goal for the FCOs continued to be met as the fish community remained diverse, self-regulating, dominated by indigenous species, and able to support sustainable fisheries, although further rehabilitation of certain fish is required. The Lake Superior Lakewide Action and Management Plan classified all habitat indicators for Lake Superior as good. Primary production and zooplankton abundance were stable during the reporting period and unchanged from the two previous reporting periods, indicating the lower food web is healthy. Abundance of the invertebrates Mysis diluviana and Diporeia spp. were stable during the reporting period, and Diporeia spp. density exceeded target levels defined in the Great Lakes Water Quality Agreement. Lake Whitefish (Coregonus clupeaformis) abundance was lower than during the previous reporting period but was within the FCO target. Abundance of lean, siscowet, and humper forms of Lake Trout (Salvelinus namaycush) remained stable at levels seen in previous reporting periods. The FCO for non-indigenous salmonids was met as Chinook Salmon (Oncorhynchus tshawytscha), Coho Salmon (O. kisutch), and steelhead/Rainbow Trout (O. mykiss) were being sustained by natural reproduction, and their abundance remained stable or increased from previous reporting periods. The FCO for Walleye (Sander vitreus) was not met, although populations showed signs of improvement since the previous reporting period. The fish community in littoral areas and embayments continued to be diverse and composed mostly of indigenous species. No new invasive species were found in Lake Superior during the reporting period. Degraded embayment and tributary habitats continued to prevent achievement of the FCOs for Brook Trout (S. fontinalis) and Lake Sturgeon (Acipenser fulvescens). Barriers created for hydroelectric generation either blocked Lake Sturgeon from historically important spawning grounds or reduced stream flows necessary for its reproduction. In tributaries without man-made barriers, Lake Sturgeon was relatively abundant and appeared healthy. Attainment of the FCOs for Brook Trout and Lake Sturgeon will be challenging and can only be attained through development of large-scale management actions like those implemented for Lake Trout rehabilitation and Sea Lamprey (Petromyzon marinus) control. Sea Lamprey-control expenditures doubled in 2016 from past levels and should result in suppression of the population closer to its FCO after 2016. The prey-fish FCO appears to have been met, but biomass of nearly all prey-fish species declined from that reported for the previous reporting period and has been on a downward trajectory since 2000. Predation by Lake Trout is probably to blame for the declining biomass of prey fish. Poor recruitment by Cisco (C. artedi) over the last 15 years is exacerbating the declines in prey-fish biomass because Lake Trout must compensate for the loss of Cisco by consuming other, less-abundant prey fish.

Lake Superior↗

Pleistocene fishes from Alameda County, California

Six types of freshwater fishes were recovered from an early Pleistocene (Irvingtonian) locality on the east side of San Francisco Bay, Alameda County, Calif. The fauna includes one centrarchid, one salmonid, three cyprinids, and one catostomid. The fauna indicates fluvial and slow-moving or lacustrine aquatic environments. One of the cyprinids and the catostomid are assigned to provisional genera because of the inadequate amount of material available for study and the obvious differences between these fossils and known fossil or recent fishes. They are new additions to the Pleistocene fish fauna of California. Paleogeographic distribution of some of these fishes indicates former fluvial or lacustrine connections between Utah, Idaho, Oregon, Washington, Nevada, and California. The Pliocene and Pleistocene fossil fish faunas indicate the widespread occurrence of genera now endemic to California. A similar picture is presented by the molluscan evidence.

California↗