This project combines two previously independent steelhead kelt contracts (2000-017-00 & 2003-062-00).  This project is a collaborative study to investigate approaches to increase adult steelhead returns by utilizing the kelt life stage.  Approaches range from low intensity/cost such as collect and transport kelts, to high intensity/cost methods including holding and feeding for kelts for several months.  An additional directive of the project, which was the focus of project 2003-062-00, is to directly evaluate relative reproductive success of three variants of steelhead Oncorhynchus mykiss (natural-origin, hatchery-origin, and reconditioned kelt) across two ESUs of interest as listed in the BPA Request for Studies released in 2003.  The nature of this work dictates long-term commitments to funding and monitoring.  As proposed this project will operate for 12 years, thus encompassing 2 steelhead generations and 1 reference year-class.  The goal of the project is to develop novel steelhead restoration approaches.

The expression of iteroparity is apparently depressed in steelhead populations that spawn in tributaries located upstream of Bonneville Dam, due primarily to the hydrosystem.  Large numbers of kelt steelhead are observed at collector dams like Lower Granite Dam, but abundance of those same steelhead at Bonneville Dam is relatively small.  

The recent ESA listing of many Columbia Basin steelhead populations has prompted interest in developing reconditioning methods for wild steelhead populations within the Basin.  To address recovery and reinstate this valuable life history strategy, the Yakama Nation in collaboration with the Columbia River Inter-Tribal Fish Commission (CRITFC) began capturing wild emigrating kelt steelhead from the Yakima River in 1999 to test reconditioning and the effects of several diet formulations on its success at Prosser Hatchery on the Yakima River (BPA Project 200001700).  The reconditioning project has developed techniques to successfully recondition kelt steelhead, but prior to widespread implementation of the procedures, the Independent Scientific Review Panel (ISRP) suggested that a study evaluate the relative reproductive success of artificially reconditioned kelt steelhead.  

This evaluation program is designed to investigate the reproductive success of hatchery-reared, natural-origin, and reconditioned kelt steelhead Oncorhynchus mykiss in two different evolutionary significant units (Upper Columbia and Middle Columbia) under natural conditions.  The two major goals are 1) directly examine reproductive success in two streams; and, 2) replicate and evaluate kelt reconditioning procedures and protocols at a variety of locations.  This project is a collaborative effort among three tribes (Warm Springs, Yakama Nation, and Colville), the University of Idaho, and the Columbia River Inter-Tribal Fish Commission.

We will apply kelt reconditioning methods developed by project 2000-017-00 with geographic replication.  Direct examination of reproductive success will be accomplished using pedigree analysis.  At all of our study sites returning adults of all variants (hatchery-origin identified by missing adipose fin, natural-origin identified by intact adipose fin, and reconditioned kelt collected from the previous spawning migration) will be DNA-typed in order to establish parentage of juveniles sampled from rearing areas above the weir.  A total of 10 to 12 microsatellite loci will be assayed using nonlethal tissue sampling methods.  Additionally, we will also collect and DNA-type approximately 200 adult resident rainbow trout O. mykiss at each stream in an attempt to identify parentage from resident forms.  We also plan to implant PIT tags in each fish (resident and anadromous forms) sampled to track their migration through the hydrosystem.  

If relative reproductive success is high, reconditioning kelt steelhead could provide a means for maintaining this natural life history characteristic and potentially aid in the recovery of listed stocks.  This study will provide resolution on uncertainty and genetic risk associated with the use of artificial propagation and reconditioning kelts in recovery of listed populations.

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.  Three 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.  Major coordination tasks include:

2.  Evaluate reproductive success of natural-origin, hatchery-origin, and reconditioned kelt steelhead and adult resident O. mykiss at Shitike Creek, and Omak Creek using pedigree analysis.

Rationale:  Determining reproductive success of individual steelhead involves very intense monitoring and evaluation.  In addition, regulation of access to the spawning area must be maintained.  Monitoring efforts will involve trapping adults and juvenile steelhead and tagging individuals with PIT tags.  Tissue samples will be collected from all adult individuals and from a random sample of juveniles.  

Highly polymorphic microsatellite loci have become the marker of choice for parentage and population studies due to the potential for differentiating closely related populations and accurate parentage assignment.  Utilizing microsatellite loci optimized for steelhead studies, we plan to determine the reproductive success of wild, hatchery, and kelt steelhead from three replicate sites in the Columbia Basin.  The process will be comprised of four steps:  1) collect year 2006 samples from each of three study sites (all adult returns over three selected weirs, smolt progeny from screwtraps, and adult resident rainbow trout),  2)  generate microsatellite genotypes from all samples taken in 2006 and perform parentage assignments, 3) collect adult returns of brood year 2006 steelhead (annually in 2008, 2009, and 2010), and 4) generate microsatellite genotypes of annual adult returns and assign parentage.  Specifically, we will attempt to assign the parentage of juvenile progeny (and subsequent adults) back to adult collections of wild, hatchery, or kelt steelhead.  This method will allow us to quantify not only the reproductive contribution of individual fish, but also quantify the adult returns related to each parental category of steelhead.

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 of the three study sites, kelt steelhead will be collected as they accumulate on the upstream side of each picket weir.  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.
  
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.  This further refines fish culturing techniques to emphasize rematuration of steelhead not just individual survival.  The majority  of this work is performed by the University of Idaho under subcontract.