Short Description

This project is a collaborative approach 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.  Additionally, the project will evaluate kelt steelhead reconditioning and rematuration rates spatially and temporally.  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.

Background

The expression of iteroparity is apparently depressed in steelhead populations that spawn in tributaries located upstream of Bonneville Dam, due primarily to the hydrosystem (Evans and Beaty 2000, 2001; Wertheimer et al. 2002; Hatch et al. 2003b).  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 trait, the Yakima/Klickitat Fisheries Project (YKFP) in collaboratively 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 kelts reconditioned during that project substantially bolstered the number of repeat spawners in the study streams.  Valuable knowledge regarding kelt husbandry, food type preferences, condition, and rearing environments have been obtained during the research endeavor.  Since the project's inception in 1999, 20-30% of the kelts collected annually have been successfully reconditioned, and radio telemetry provided the ability to track some of these fish to the spawning grounds and to obtain documentation of successful redd construction.  In terms of numbers of fish, this means that an additional 100-200 steelhead females were available to spawn a second time (total potential of approximately 300,000-600,000 eggs at an estimated 3,000 eggs per female) in the Yakima Basin that, without this program, would likely not have been able to do so.  Total egg production estimates are impressive, however, the reproductive success of these reconditioned kelt steelhead is unknown and is part of the basis for this study.

In the Yakima River kelt study, Passive Integrated Transponder (PIT) tags are also being used to track captured steelhead kelts through the reconditioning process.  An example from the PIT tag history of two such female kelts demonstrates the promise of this program to substantially increase the number of repeat-spawning steelhead in the Yakima River Basin (Table 1).  Data from the second female demonstrate that the reconditioning process apparently did not affect this kelt's ability to home to that portion of the Yakima River Basin from which it originated Information collected during this feasibility study has been significantly incorporated into the experimental design for upcoming years of research, and is expected to continue to increase survival and successful expression of repeat spawning.  For a detailed report on BPA project 200001700 see Hatch et al. (2003a).


Table 1.
Mark/recapture data for two PIT-tagged Yakima River steelhead kelt females captured and reconditioned at the Prosser Hatchery in 2001 and 2002.

Description Location Date Post-eye-to-hyperal         Weight (kg)
3D9.1BF11A2C6A          length (cm)

Captured / tagged / Prosser 10-May-01 45 1.3
reconditioned Prosser 15-Nov-01 52 3.0
Released Prosser 30-Apr-02 51 1.9
Recaptured / reconditioned Prosser 10-Dec-02 53 2.3
Released

3D9.1BF1456C7D
Captured / tagged Roza 2-Apr-02 47 2.1
Recaptured  / reconditioned Prosser 25-Apr-02 48 1.7
Released Prosser 10-Dec-02 52 3.7
Recaptured / released Roza 7-Mar-03 53 3.5

General Approach

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 three 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 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,
     Omak Creek, and a tributary of the Yakima River 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 (Bernatchez and Duchesne 2000; Eldridge et al. 2002; Estoup et al. 1998; Letcher and King 2001; Norris et al. 1999; O'Reilly et al 1998).  Utilizing microsatellite loci optimized for steelhead studies (Narum et al. in review), 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 2005 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 2005 and perform parentage assignments, 3) collect adult returns of brood year 2005 steelhead (annually in 2007, 2008, and 2009), 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 (WE 11).  Anesthetized steelhead will be visually examined to classify each fish as a kelt or prespawn individual.  Methods for visual classification are available (Hatch et al. 2003b) and primarily involve keying specimens based on an imploded abdomen.  This visual technique was highly precise when compared with the use of ultrasound analysis (Evans and Beaty 2001).  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" (WE 12), 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.  Transporting the kelt steelhead is covered under WE 13.  Work elements below will cover kelt rearing/reconditioning, release back into the stream of capture, and comparison of reconditioning success among sites.  

In-Kind Contributions

The CRITFC genetics laboratory in Hagerman, ID works collaboratively with the University of Idaho and the U.S. Department of Agriculture.  All three groups share laboratory equipment housed at the U of I Experiment Station.  A Memorandum of Agreement governs the arrangement.  The scientists cooperatively share equipment and the genetic sequencer that is being leased under this contract with BPA will be used in the same manner.  In-kind contributions of equipment for this project are detailed below:

Item Estimated value
DNA extraction robot $65,000
Thermalcyclers (five) $35,000
Liquid handling robot $85,000
Freezers (three $36,000
Cooling unit 4deg C $5,000
Ice machine $2,000
Plate centrifuge (two) $40,000

References

Bernatchez, L. and P. Duchesne. 2000. Individual-based genotype analysis in studies of parentage and population assignment: How many loci, how many alleles? Canadian Journal of Fisheries and Aquatic Sciences 57:1-12.
Eldridge, W. H., M. D. Bacigalupi, I. R. Adelman, L. M. Miller, and A. R. Kapuscinski. 2002. Determination of relative survival of two stocked walleye populations and resident natural-origin fish by microsatellite DNA parentage assignment. Canadian Journal of Fisheries and Aquatic Sciences 59:282-290.
Estoup, A, K. Gharbi, M. SanChristobal, C. Chevalet, P. Haffray, and R. Guyomard. 1998. parentage assignment using microsatellites in turbot (Scophthalmus maximus) and rainbow trout (Oncorhynchus mykiss) hatchery populations. Canadian Journal of Fisheries and Aquatic Sciences 55:715-725.

Evans, A.F., and R.E. Beaty.  2000.  (DRAFT) Identification and enumeration of steelhead (Oncorhynchus mykiss) kelts at Little Goose Dam juvenile bypass separator, 1999.

Evans, A.F. and R.E. Beaty.  2001.  Identification and enumeration of steelhead (Oncorhynchus mykiss) kelts in the juvenile collections systems of Lower Granite and Little Goose dams, 2000.  Ann. Rep. To US Army Corps of Engineers, Walla Walla District, for Contract No. DACW-00-R-0016. Prepared by the Columbia River Inter-Tribal Fish Commission, Portland OR.

Gerber S., Chabrier P., Kremer A. (2003) FaMoz: a software for parentage analysis using dominant, codominant and uniparentally inherited markers, Molecular Ecology Notes, in press.

Hatch, D.R., R. Branstetter, J. Blodgett, B. Bosch, D. Fast, and T. Newsome. 2003a.  Kelt reconditioning: A research project to enhance iteroparity in Columbia Basin steelhead (Oncorhynchus mykiss).  Annual Report to the Bonneville Power Administration for Contract No. 00004185.  Prepared by the Columbia River Inter-Tribal Fish Commission, Portland OR.

Hatch, D.R., R. Branstetter, and S. Narum. 2003b.   Evaluate steelhead (Oncorhynchus mykiss) kelt outmigration from Lower Granite Dam to Bonneville Dam and test the use of transportation to increase returns of repeat spawners.  Annual Report to US Army Corps of Engineers, Walla Walla District, for Contract No. DACW68-00-C-0027.  Prepared by the Columbia River Inter-Tribal Fish Commission, Portland OR.

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Letcher B.H. and T.L. King. 2001. Parentage and grandparentage assignment with known and unknown matings: application to Connecticut River Atlantic salmon restoration. Can. J. Fish. Aquat. Sci. 58(9): 1812?1821.

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Wertheimer, R.H., P.L. Madson, M.R. Jonas, and  J.T. Dalen.  2002.  Evaluation of steelhead kelt project abundance, condition, passage, and conversion rates through lower Columbia River dams, 2001.  Report of Monitoring US Army Corps of Engineers, Portland District Fisheries Field Unit, Bonneville Lock and Dam, Cascade Locks, OR.