BPA Fish and Wildlife FY 1998 Proposal

Section 1. Summary
Section 2. Goals
Section 3. Background
Section 4. Purpose and methods
Section 5. Planned activities
Section 6. Outcomes, monitoring and evaluation
Section 7. Relationships
Section 8. Costs and FTE

see CBFWA and BPA funding recommendations

Section 1. Summary

Title of project
Listed Stock Gamete Preservation

BPA project number   9703800

Short description
The Listed Stock Gamete Preservation project seeks to apply cryogenic technology to preserve male gametes from chinook salmon conservation units that are at low levels of abundance and high risk of extirpation. This approach would target chinook salmon populations with ongoing conventional hatchery or captive broodstock artificial propagation programs to preserve and utilize cryopreserved material to enhance genetic diversity. Secondly, this project would establish a gene bank as an insurance policy in case that extirpation of chinook salmon populations does occur. Cryopreservation for hatchery steelhead trout programs that use native brood sources could also be included if desired.

Business name of agency, institution or organization requesting funding
Nez Perce Tribe Department of Fisheries Resources management

Proposal contact person or principal investigator

 NamePaul A. Kucera, Director of Biological Services
 Mailing address
Lapwai, ID 83540
 Phone208/843-7320 x2

University of Idaho; Washington State University

Section 2. Goals

Maintains biological diversity; maintains genetic integrity; when utilized with hatchery propagation efforts it could enhance genetic diversity of the propagated population and when gene banked allows for future management and research options.

Target stockLife stageMgmt code (see below)
Lostine River chinookAdultL, S, W
Upper Wallowa River chinookAdultL
Minam River chinookAdultL, N, d
Wenaha River chinookAdultL, N
Upper Grande Ronde R. chinookAdultL, S
Imnaha River chinookAdultL, S
Big Sheep and Lick Cr. chinookAdultL, S
Snake River mainstem trib. chinookAdultL,
Lower Salmon River mainstem trib. chinookAdultL
Little Salmon River trib. chinookAdultL
Rapid River chinookAdultL, N
Upper mainstem S. Fk. Salmon River hatcheryAdultL, S
Mainstem S. Fk. Salmon R. summer chinookAdultL, N
Secesh River chinook (includes Lake Creek)AdultL, N
Johnson Creek chinookAdultL, N
SEast Fork South Fork Salmon chinookAdultL, S, d
Marsh Creek and Trib. chinookAdultL, N
Bear Valley/Elk Creek chinookAdultL, N, d
Sulphur Creek chinookAdultL, N
Loon Creek chinookAdultL, N
Camas Creek chinookAdultL, N
Big Creek chinookAdultL, N, d
Lemhi River chinookAdultL, S
Pahsimeroi River chinookAdultL, S
North Fork Salmon RiverAdultL, N
E. Fk. Salmon, mouth to Herd Cr. chinookAdultL
Herd Creek/upper E. Fk. chinookAdultL
Yankee Fork and trib. chinookAdultL, S, d
Valley Creek chinookAdultL
Mainstem Salmon River below Redfish Lk. Cr.AdultL, S
Mainstem Salmon River above Redfish Lake Cr.AdultL, S


Section 3. Background

Stream area affected

Stream name   All streams and chinook salmon populations listed in NMFS Proposed Recovery Plan for snake River salmon (1995).
Hydro project   The NPPC (1994) directs the Council to develop a program to protect, mitigate and enhance fish and wildlife on the Columbia River and its tributaries ...... affected by the development, operation and management of [hydroelectric projects].......
Subbasin   Middle Fork Salmon River; upper Salmon River; Lemhi River; Pahsimeroi River; South Fork Salmon River; mainstem Salmon River tributaries; Grande Ronde River; Imnaha River; and Snake River tributaries.
Land ownership   Both.

The Listed Stock Gamete Preservation project has previously received a relatively high project ranking through the Columbia Basin Fish and Wildlife Authority. This project was funded in FY 1997 for the first time. The Lower snake River Compensation Plan hatchery evaluations program funded through the U.S. Fish and Wildlife Service has provided a valuable but limited amount of support for this effort since 1992.

Biological results achieved
None to date. Project to be initiated in 1997.

Project reports and papers
None to date. Project to be initiated in 1997.

Adaptive management implications
This project would provide cryopreserved male gametes for ongoing chinook salmon artificial propagation production programs to enhance genetic diversity in those listed chinook salmon populations. It would allow for preservation of genetic material from salmon populations at low levels of abundance and high risk of extirpation. A germplasm repository would be developed for this purpose as an insurance policy against potential extirpation of the species and to provide for future management and research options. The chinook salmon captive broodstock plan (ODFW 1996) for Grande Ronde salmon populations recognizes the importance of and contains guidelines for the use of cryopreserved semen to maintain genetic diversity of the propagated populations.

Section 4. Purpose and methods

Specific measureable objectives
This project proposes to utilize cryogenic technology to gene bank male salmon gametes from listed snake River chinook salmon conservation units at low levels of abundance and high risk of extirpation. A germplasn repository would eventually be established that would allow safe preservation and storage of cryopreserved germplasm, that would be available to use in conventional hatchery and captive broodstock programs. Material would also be preserved and available for future use. A specifc measurable objective would be that cryopreserved material used in artificial propagation programs would result in quantifiable fertilization rates and measurable genetic profiles.

Critical uncertainties
See Underlying Assumptions or Critical Constraints.

Biological need
Snake River spring/summer chinook salmon populations have experienced significant decline in population numbers over the past five decades and are now listed as a threatened species. Genetic conservation through population protection and monitoring has not been successful. A genetic resource management approach using cryogenic techniques is recommended for preservation of genetic material from salmon conservation units at high risk of extirpation. These efforts will be needed to preserve and maintain genetic diversity for ongoing artificial propagation programs and to preserve genetic material for future management options. The NMFS draft recovery plan states that "captive broodstock and supplementation programs should be initiated and/or continued for populations identified as being at imminent risk of extinction, facing severe inbreeding depression, or facing demographic risks". The plan further states that"the conservation of local populations or stocks of Pacific salmon and the preservation of their genetic resources is an important goal".

Hypothesis to be tested
Not research oriented.

Alternative approaches

Justification for planning

Existing weirs that trap listed adult Snake River spring/summer chinook salmon for artificial propagation will be utilized as sources of male salmon for cryopreservation purposes. Hatchery managers will be coordinated with to determine acceptable periods for sampling to occur. Streams that have had captive broodstock juvenile chinook salmon collected from them will be sampled for collection of wild/natutal adults as will other streams where gene banking will occur. Current limitations include observed fertilization rates of from 0 to 80%. The highest average fertilization rates have been approximately 65% (Mendel - Wash. Dept. of Fish and Wild. - pers. comm.). Mendel also suggested that same day cryopreservation of fresh semen seems to provide much higher fertilization rates when using cryopreserved semen. Cryogenic preservation of female gametes is also a desired product. However, the technology to successfully cryoporeserve female gametes or embryos is currently not available to fisheries science.

Standardized fish sampling, fish handling and fish anesthesia protocols, consistent with Section 10 permits, will be used to collect and handle adult male chinook salmon for semen cryopreservation purposes. Handling and sampling at hatchery traps would utilize IDFG or ODFW protocols on normally scheduled spawning days. We would also rewquest that male salmon be held as long as possible after spawning so that additional samples may be collected. Abdomens of anesthetized males will be wiped dry and semen samples will be taken into two separate whirl paks. After semen samples are taken adult recovery and semen sample storage procedures occur simultaneously. Adult salmon are immediately tagged, lengths and scales taken, data recorded and the fish placed in a holding pond or pool area where it is assisted until fully recovered. Fin punches are takne and preserved in labelled containers for later DNA extraction. Semen samples are placed in two individually labelled whirl paks, oxygenated, placed in a covered insulated cooler on wet ice on top of newspaper and transported to the University of Idaho (UI), Moscow, ID, and Washington State University (WSU), Pullman, WA. Cryopreservation occurs onsite or independently at the two Universities within a 24 hour period after the samples are taken. Samples are stored in four liquid nitrogen canisters at the UI and WSU.

Section 5. Planned activities

Phase PlanningStart 3/97 End 6/97Subcontractor NA.
1. Task 2. Task 3. Coordinate gamete preservation activities with state, federal and Tribal management agencies. Determine chinook salmon populations for germplasm sampling, including sample sizes and number of years sampling will occur. Coordinate with NMFS for necessary Section 10 scientific permits through NMFS. STATEMENT OF WORK CONTAINS OBJECTIVES AND TASKS.
Phase ImplementationStart 7/97 End Minimum of 5 years (TBD).Subcontractor University of Idaho; Washington State University.
1. Sampling of adult male salmon would occur at existing chinook weirs used for artificial propagation purposes. Cryopreservation samples would be collected from approximately July to mid-September at these weir locations. Stream sampling would occur mainly in August and September. Database and sample inventory development would take place from September to December. Report preparation would occur from November through December of each year. Project implementation would continue until a sufficient number of unrelated samples were collected from each population (minimum anticipated project term of five years). STATEMENT OF WORK CONTAINS PROJECT OBJECTIVES AND TASKS.
Phase O&MStart 1997 End Annual.Subcontractor University of Idaho; Washington State University.
1. Storage and maintenance of cryopreserved genetic material will be ongoing after implementation in the summer of 1997. The germplasm repository for the short-term has been identified as the University of Idaho and Washington State University.

Constraints or factors that may cause schedule or budget changes
Annual abundance in each chinook salmon population. Timing of sampling to collect only spawned-out males. See Underlying Assumptions or Critical Constraints section.

Section 6. Outcomes, monitoring and evaluation


Expected performance of target population or quality change in land area affected
Preservation and use of male chinook gametes for use in artificial propagation programs to maintain and enhance genetic diversity. Establishment of gene banks for preservation of genetic material from salmon conservation units at high risk of extinction.

Present utilization and convservation potential of target population or area
Almost all of the listed Snake River chinook populations are below threshold numbers of spawning adults in each population/stream and are below 10% of their historical production potential. Some populations such as the upper Big Creek population has experienced three consecutive years of cohort collapse (2 redds in 1994, 1 redd in 1995, and 1 redd in 1996) and is imminent danger of extirpation. This project has the potential for genetic conservation of the male portion of the Big Creek, and other, salmon populations through application of cryogenic technology and preservation for future use. Currently, none of the cryopreserved material has been utilized in artificial propagation programs.

Assumed historic status of utilization and conservation potential

Long term expected utilization and conservation potential for target population or habitat
The desired long term conservation would be to maintain the genetic diversity of the propagated salmon population and wild populations. We would prefer to manage for healthy, self sustaining populations of snake River chinook salmon that would not require the use of cryopreserved genetic material.

Contribution toward long-term goal
Genetic material can be used in concert with artificially propagated salmon populations to maintain genetic diversity and the populations ability to change with a changing environment. Cryopreserved material would also be available for future management and research use.

Indirect biological or environmental changes

Physical products
Cryopreserved chinook salmon semen wil be stored in 0.5 ml and 4 ml straws in tanks containing liquid nitrogen.

Environmental attributes affected by the project

Changes assumed or expected for affected environmental attributes

Measure of attribute changes

Information products
Annual reports will be developed and presented for regional publication and dissemination.

Coordination outcomes
This project will be coordinated with the state, federal and Tribal salmon managers. coordination should ensure collection of male salmon gametes from existing listed stock hatchery programs.

Goals will be developed and refined through multi-agency coordination to determine which populations should be addresed and how mny samples should be cryopreserved over the next five year period. Project performance, in terms of number of samples preserved annually, can be easily monitored. Genetic profiles of conventional hatchery reared chinook populations and populations from captive broodstock programs that utilize cryopreserved semen can also be folowd to determine if natural and hatchery populations are imilar or dissimilar.

Provisions to monitor population status or habitat quality
Monitoring population status is not a goal of his project. Tribal, state and federal agencies conduct annual chinook salmon redd count surveys and escapement monitoring studies to document the relative abundance of individual salmon populations. Genetic analysis has been conducted by NMFS to document population profiles and follow them over time. This project will keep an updated computer inventory of all available chinook salmon semen.

Critical uncertainties affecting project's outcomes
Additional research needs to be conducted by experts on cryopreservation mehodology (freezing and thawing process) and the fertilization process on a much broader scale. A basin-wide cryopreservation plan should be developed by the salmon mgers.

Incorporating new information regarding uncertainties
Utilization of new information is an ongoing process that allows project refinement and adaptive management to take place.

Increasing public awareness of F&W activities
This project seeks to preserve genetic material from threatened spring/summer chinook salmon populations from the snake River and to establish a germplasm repository or the same. It is not an information and education program, perse, and is only a stop-gap measure until improvement in minstem survivl conditions alow for self-sustaining populations of salmon.

Section 7. Relationships

Related BPA projectRelationship
14-48-301-97502 Lower Snake River Compensation Plan McCall Hatchery, Lookingglass Hatchery and Sawtooth Hatchery
9604300 Presereved genetic material may be used in spawning protocols to promote genetic diversity.
9604400 Grande Ronde Basin Spring Chinook Captive Broodstock Program Capitol Construction Component Preserved genetic material may be used in spawning protocols to promote genetic diversity.
Related non-BPA projectRelationship
Lower Snake River Compensation Plan Nez Perce Tribe Evaluation Studies - 1997This project currently provides limited funds and personnel to collect and cryopreserve male gametes of listed Snake River chinook salmon from selected streams.

Opportunities for cooperation
This project would depend on cooperation of those agencies that currently operate adult chinook salmon trapping facilities; mainly IDFG and ODFW. Adult chinook salmon semen was collected in a cooperative effort with IDFG at the South Fork Salmon River fish weir and at Lookingglass Hatchery (Imha River adults) with ODFW in 1996.

Section 8. Costs and FTE

Future funding needs   Past obligations (incl. 1997 if done)
FY$ Need% Plan % Implement% O and M
1998140,000 0%90% 10%
1999145,000 0%90% 10%
2000150,000 0%90% 10%
2001155,000 0%90% 10%
2002155,000 0%90% 10%

Other non-financial supporters
U.S. Fish and Wildlife Service, Lower snake River Compensation Plan program; Washington Department of Fish and Wildlife; Oregon Department of Fish and Wildlife.

Longer term costs   It is anticipated that cryopreservation of listed hatchery populations may continue past 2002 if preserved samples are utilized on an annual basis. In adition, if sufficient samples cannot be collected from slmon populations because of low abundance, additional years collections would occur. Operation and mintence of the cryopreserved germplasm would need to occur at some level into the future. Costs are uncertain at this time.
FY97 overhead percent   29.5%

How does percentage apply to direct costs
Indirect rates apply to a portion of the direct costs, excluding equipment and services.

Subcontractor FTE   0.08 FTE - University of Idaho; 0.08 FTE - Wash. State University.