The Kelt Steelhead Reconditioning and Reproductive Success Evaluation Project is a research, monitoring, and evaluation (RM&E) category project funded through the Columbia Basin Fish Accords.  The project studies and evaluates two broad topics with respect to post-spawn steelhead, first it assesses reconditioning processes and strategies, and second, it measures reproductive success of artificially reconditioned kelt steelhead.  It associates with RPAs 33 and 64 in the Federal Columbia River Power System Biological Opinion.  RPA 33 requires the Action Agencies to develop, in cooperation with regional salmon managers, and to then implement a Snake River steelhead kelt management plan designed to provide at least a 6% improvement in B-run population productivity.  Toward that goal, a variety of approaches are being tested and implemented including passage improvements and reconditioning kelt stage steelhead.  This project focuses on the reconditioning component.  RPA 64 involves resolving artificial propagation critical uncertainties primarily through relative reproductive success studies.  This project is working toward evaluating reproductive success of artificially reconditioned kelt steelhead. This research contributed three papers to the published literature in 2013 (Caldwell et al. 2013; Hatch et al. 2013; Penney and Moffitt 2013) and an additional four papers are in review (Buelow et al. in review; Caldwell et al. in press; Penney and Moffitt in review a.; Penney and Moffitt in review b.).  Our team presented 14 project presentations in 2013 at basin, regional, national, and international levels.  

Reconditioning Processes Strategies

Kelt steelhead reconditioning process evaluations involve fish culturing practices, studying alternative management strategies, and implementing research scale reconditioning programs.  Refinements in fish culturing practices this year included a diet experiment that investigated the benefits of top dressing pellets with fish oil or cyclopeeze, and evaluating the effectiveness of ivermectin and emamectin benzoate for parasite control.  Effectiveness was measured with survival and rematuration metrics.  We used fish in long-term reconditioning for a feeding trial that compared an “orange” diet that was comprised of a mix of customized Bio-oregon brood pellets top coated with a mix of cyclopeeze and Alaskan fish oil versus the standard diet (customized Bio-oregon diet and krill) (Hatch et al 2003a). Both groups received krill at the start of reconditioning for the same duration of time. The diet study just recently concluded in 2013 and data is currently being analyzed. Preliminary results found in the 2012 trial, fish fed the orange diet had higher muscle lipid levels at release than fish fed the standard diet. The orange diet median muscle lipid levels increased by 0.4% in 2013, and 0.6% in 2012.  The increase in muscle lipid levels was modest and may be explained due to either increased feed consumption or the higher lipid level in the orange diet. If feasible, the current feeds should have increased lipid content and appetite stimulants added before further diet studies are undertaken.  Emamectin benzoate (emamectin) is an alternative to ivermectin that has been developed for control of sea lice (marine copepod ectoparasites) in Atlantic salmon aquaculture, and both drugs have been adapted for use in steelhead kelt reconditioning to reduce Salmincola is a genus of parasitic copepod that can inhibit oxygen uptake and gas exchange at the gill lamellae/water surface interface.  We compared the post treatment mortality rates of kelts that were administered ivermectin via gavage as well as a group that was injected. Results demonstrated a dramatic reduction in mortality that was substantial and consistent between years: after 90 days, mortality in emamectin treated fish was 33.2% less than ivermectin treated fish in 2011, and 34.8% in 2012. This experiment did not attempt to quantify levels of copepod infestation in emamectin versus ivermectin treated fish. However, very few fish were observed with significant numbers of copepods at release, suggesting that both ivermectin and emamectin treatment controlled copepods adequately.

Plasma estradiol was extracted and analyzed from long-term reconditioned kelt steelhead to assess rematuration levels.  Samples are currently being processed in the laboratory so the final results are not yet available.
  
Alternative management strategies for kelt steelhead studied thus far include: long-term reconditioning, where fish are collected during out migration, held and feed through the summer and released in the fall; collect and transport kelt steelhead, where fish are collected and immediately transported (unfed) and released below Bonneville Dam, or collected, fed for 4-6 weeks (fed) then transported and released below Bonneville Dam; and direct release, where fish are collected, PIT tagged and released back to river as a control group.

Experimental scale long-term kelt reconditioning survival data was compared from 3 locations including; the Okanogan, Snake, and Yakima rivers, where survival rates in long-term reconditioning was 8.2%, 51.8%, and 48.7% respectively.  We also published a manuscript detailing the long-term kelt reconditioning program at Prosser Hatchery (Hatch et al. 2013b).  

Specific to Snake River B-run steelhead (addressing RPA 33), our collection locations included kelts at the Lower Granite Dam juvenile bypass separator, air-spawned steelhead being used for the South Fork Clearwater River naturalized broodstock project, and Dworshak National Fish Hatchery returns.  Fish collected from Lower Granite Dam were either transported to Dworshak Hatchery for reconditioning, or PIT tagged and released back into the Snake River is represent a control group.  Fish collected from the South Fork Clearwater were reconditioned and released, and Dworshak Hatchery fish were used for experiments and not released.  Kelt stage steelhead were collected at Lower Granite Dam as they passed through the juvenile bypass system.  A total of 110 wild female B-run steelhead were collected at Lower Granite Dam from April to June and transported to Dworshak National Fish Hatchery for reconditioning.  Fifty-seven of these fish survived reconditioning and were released downstream of Lower Granite Dam in October 2013.  Additionally, 24 B-run kelt steelhead were retained for reconditioning after air spawning at Dworshak Hatchery.  These fish were part of project that is developing localized broodstock for the South Fork Clearwater River.  Following reconditioning 12 of these fish were released back into the South Fork Clearwater River.  In total 69 wild B-run steelhead were reconditioned and released back into the Snake River system aimed at addressing RPA 33.  

We did not conduct any transport experiments in 2013 due to their relatively lower benefits compared to long-term reconditioning and the overall lower availability of kelt steelhead.  From 2002 through 2011 we collected and transported kelt steelhead from Lower Granite Dam to below Bonneville Dam.  These studies collected and transported 2,698 kelt steelhead, of which 29 (0.01%) were detected returning upstream at Bonneville Dam.

We are developing a spreadsheet model of management scenarios to address “what if” simulations.  This model will be completed during the first half of 2014.

To establish a control group for management action comparisons, we continue to collect and PIT tag the kelt steelhead run at large to estimate baseline in-river survival.  In 2013, we tagged 827 steelhead collected from the Lower Granite Dam juvenile bypass system.  None of these fish were detected returning at any FCRPS dam as sequential spawners in 2013.  It is possible that some fish from this release will exhibit a skip spawner life history pattern and return in 2014 after spending an additional winter in the ocean.  Similar tagging in 2012 resulted in the return of a single fish out of 2,098, resulting in a return rate to Bonneville Dam of 0.05%.  Since 2002, we have PIT tagged 9,325 kelt steelhead at Lower Granite Dam and detected 32 (0.34%) of them returning upriver at Bonneville Dam.  The 2013 Yakima River control group release included 52 PIT tagged fish.  None of these fish were detected returning to Bonneville Dam as sequential spawners.  From 2005 through 2013, 669 kelt steelhead were PIT tagged and released in the Yakima River representing the control group and so far 21 (3.4%) fish were detected returning to Bonneville Dam.

The genetic stock structure of Snake River Basin kelt steelhead was examined using 187 SNP markers extracted from emigrating fish collected at Lower Granite Dam.  The kelt steelhead population composition at the Lower Granite Dam Juvenile Bypass Separator from 2009-2012 was Upper Salmon (0.39); Grande Ronde/ Lower Snake (Tied) (0.16); Middle Fork Salmon (0.09); and, Imnaha (0.06), reporting groups based on a sample size of 4,138.

Reproductive Success of Artificially Reconditioned Kelt Steelhead

We have investigated reproductive success of artificially reconditioned kelt steelhead at a variety of scales.  Experiments have been conducted at the natural stream level, in a hatchery environment, and at the individual fish level.
This evaluation program is designed to investigate the reproductive success of artificially reconditioned kelt steelhead.  Since direct examination of reproductive success in the field has proven very difficult, we are concentrating on measuring physiological and endocrinological parameters as an index to rematuration and reproductive success.  Additionally, we are evaluating gamete and progeny viability and conducting reproductive success studies in Omak Creek using pedigree analysis.   This project is a collaborative effort among four tribes (Nez Perce, Warm Springs, Yakama Nation, and Colville), the University of Idaho, and the Columbia River Inter-Tribal Fish Commission.  

At the natural stream level, we have conducted experiments on Omak Creek a tributary to the Okanogan River for several years.  We have determined that at least 4 of 11 reconditioned kelt steelhead have successfully produced offspring in Omak Creek.  This site has proven challenging for this evaluation primarily due to low kelt steelhead collection numbers and then ultimately few reconditioned fish.  Additionally, the reconditioned fish must be released in the Okanogan River to overwinter adding mortality and difficulty to tracking these fish to spawning grounds in Omak Creek.  An additional 22  juveniles assigned to reconditioned kelts in the Yakima River.  As a result of these challenges, we may transfer future efforts to the Cle Elem Hatchery spawning channel (described below).  

In 2013, we radio tagged 70 long-term reconditioned kelt steelhead prior to release in the Yakima River.  These fish were known rematuring based on elevated levels of estradiol.  We are partnering with the Yakima River Steelhead VSP project to track these fish to spawning beds.  Later we will target electrofishing efforts downstream of known reconditioned kelt spawners in an attempt to identify their progeny.  In addition, 21 of these radio tagged fish have known detection histories from prior spawnings.  These histories are from in-stream PIT tag detection or from previous radio tags.  Relocating these fish will provide data on repeat homing fidelity of artificially reconditioned kelt steelhead.

At Parkdale Fish Facility, we conducted experiments to compare performance of maiden and repeat spawnings of artificially reconditioned kelt steelhead using metrics of fecundity, fertilization rates, and early juvenile growth.  A total of 21 fish were successfully reconditioned and repeat spawned from a collection of 134 steelhead.  Summer-run steelhead had higher fecundity levels in their repeat compared to maiden spawning, and all other metrics showed no significant difference between repeat and maiden spawnings.  Winter-run steelhead showed no significant differences in fecundity, fertilization rates, and growth in terms of length and weight, there were significant differences in fry starting length and weight with maiden spawnings resulting in larger fry starting sizes. In 2014 we are conducting similar work on B-run steelhead at Dworshak National Fish Hatchery.

To better understand the physiological processes that influence post-spawn steelhead recovery, we published two manuscripts, the first used histological analysis of Snake River steelhead as a model to assess the cellular architecture in the pyloric stomach, ovary, liver, and spleen in sexually mature and kelt steelhead (Penney and Moffitt 2013). The second identified metabolic endocrine factors involved in spawning recovery and rematuration of iteroparous female rainbow trout (Caldwell et al. 2013).

From an adaptive management standpoint in 2014, the experimental spawning channel at Cle Elum Hatchery will be available, so we are currently drafting a proposal and study plan to use the channel in an evaluation of reconditioned steelhead reproductive success.  Additional components such as resident rainbow trout contribution and maiden steelhead reproductive success may also be added.

We are also searching for potential stream locations to conduct a relative reproductive success study.

Objectives

1.  Plan and coordinate all aspects of project implementation including permitting, subcontracting, and logistics.  

Rationale:  This project is very complex.  It involves geographic replication of specimen collection, artificial reconditioning of post-spawn steelhead, and state-of-the-art genetic analysis.  Two different tribal fishery staffs will be conducting field collections and CRITFC will coordinate activities.  The target species is listed under the Endangered Species Act (ESA) in all areas above Bonneville Dam, which invokes a federal permitting process.

2.  Evaluate reproductive success of artificially reconditioned kelt steelhead.

Rationale:  Determining reproductive success of individual steelhead involves very intense monitoring and evaluation.  We plan to evaluate reproductive success of artificially reconditioned kelt steelhead in the Cle Elem Hatchery spawning channel and through the Yakima River.  The spawning channel will provide a means to intensely evaluate reproduction in a controlled environment and using pedigree analysis we can assign known-age juveniles produced in the Yakima River to repeat spawning steelhead. Preliminary results from 2013 indicate, we identified 18 juvenile steelhead as progeny from reconditioned kelt steelhead in a sample 497 age 0 juvenile steelhead.  This 3.6% (18/497) component is similar to the composition that artificially reconditioned steelhead comprised in the Yakima River run at large suggesting that production contributions were similar between maiden and artificially reconditioned steelhead.  Additionally, we will study endocrinology and physiology measures that are an index to rematuration and reproduction.  This work will be concentrated in the Snake River Basin, but samples will be taken from all study areas.

3.  Apply kelt steelhead reconditioning techniques at selected streams to post-spawners for release back into study streams.

Rationale:  This objective will test the following hypothesizes:
Ho:  Kelt steelhead reconditioning rates are similar spatially and temporally; and,
Ho:  Kelt steelhead rematuration rates are similar spatially and temporally.  
Additionally this objective will provide the reproductive success experiment (objective 2) with reconditioned kelt steelhead for study.
  
At each study sites (Yakima, Snake, and Clearwater rivers), kelt steelhead will be collected as they accumulate on the upstream side of each picket weir or other collection site.  These fish will be removed with dip nets and placed in an anesthetic tank.  Anesthetized steelhead will be visually examined to classify each fish as a kelt or prespawn individual.  Methods for visual classification are available and primarily involve keying specimens based on an imploded abdomen.  This visual technique was highly precise when compared with the use of ultrasound analysis.  If a specimen is suspected to be a pre-spawner the fish will be released on the downstream side of the weir.  Following collection anaesthetized kelts will be "in-processed", where they are scanned for a PIT tags, measured, weighed, fish color and condition noted, injected with Ivomec intubate (parasite treatment), and injected with a PIT tag if not present in the specimen.  The kelts are then held in a tank prior to transport to the reconditioning facilities.  Kelt steelhead will be transported to reconditioning facilities for culturing.  After kelt rearing/reconditioning, they will be released back into the stream of capture, and comparisons of reconditioning success among sites will be made.

4.  Investigate and develop approaches to utilize the steelhead kelt life stage to increase steelhead populations.

Rationale:  Providing assistance to post-spawn steelhead in the forms of transportation, feed, and prophylactic measures may increase the probability that individual steelhead repeat spawn and contribute to population growth.  In this objective we measure the variation in steelhead response to intervention method.  Further we are attempting to identify locations that are particularly problematic to steelhead kelt migrations in the lower Columbia River and estuary.

In this objective we will study the physiology and endocrinology of steelhead kelts with a goal of evaluating the feasibility and success of several strategies for rehabilitating and handling of steelhead captured at Lower Granite Dam or at other sites during their downstream migration in the Snake River system. Our research will focus on the physiology, health and condition of both B and A stocks of steelhead trout. Through this research we will pose, develop and test protocols that can be used to collect and transport spent spawners, rehabilitate them for the most effective period of time to maximize their ability and contribution to the next spawning generation.  Our focus is to develop the background science needed for evaluating different production plans for rehabilitation of kelts. The second and third years of the study will focus on more specific studies using metrics developed in the first year as evaluation tools. The second and third year of the study will test larger groups of fish and utilized some seawater testing systems. The details and infrastructure planning aspects of this project are recognized as goals during the first year of this project.  The majority of this work will be conducted under subcontract to the University of Idaho.
  
5.  Evaluate progeny and gamete viability from  reconditioned kelt steelhead.

Rationale:  This objective is addressing the effect of artificial reconditioning on gamete and progeny development.  Work is performed on B-run steelhead at Dworshak National Fish Hatchery.  The premise is to collect hatchery-origin prespawn adults and hold them in a hatchery.  After the female fish are ripe they are air spawned, eggs are fertilized with cryopreserved milt, and the offspring are raised for several weeks while recording various measures of quality.  Female steelhead after air spawning are placed in tanks and reconditioned in a manner similar to our other long-term reconditioning treatments.  This experiment utilizes a replicated, repeated measures experimental design, to assess and compare egg and progeny viability of reconditioned vs. first time spawners.  Long-term reconditioning and subsequent captive spawning provides us with means to obtain valuable quantitative data on gonad processes, maturation rates and juvenile survival.   Data resulting from this research will greatly contribute to the evaluation of reconditioning as a conservation tool.    The hypothesis we are testing is:

Ho: Measures of gamete and progeny viability and quality are similar between first spawning and second spawning following artificial reconditioning.