Listed Stock Chinook Salmon Gamete Preservation proposal 199703800

FY07-09 proposal 199703800

Section 1. Administrative

Proposal title	Listed Stock Chinook Salmon Gamete Preservation
Proposal ID	199703800
Organization	Nez Perce Tribe
Short description	Preserve genetic diversity of endemic Snake River basin Chinook Salmon and steelhead using cryogenic technology.
Information transfer	Information will be tranfered by BPA Annual Reports published on the web, journal publications and meeting presentations. In addition, information will be shared with through meetings with the WSU/UI/NOAA Fisheries Center for Reproductive Biology, Salmon Recovery Group and the National Animal Germplasm Program, Aquatic Species Committee.
Proposal contact person or principal investigator

Contacts

Contact	Organization	Email
Form submitter
Bill Young	Nez Perce Tribe	billy@nezperce.org
All assigned contacts
Bill Young	Nez Perce Tribe	billy@nezperce.org

Section 2. Locations

Province / subbasin: Mountain Snake / Salmon

Latitude	Longitude	Waterbody	Description
		Tucannon River
		CathrineCrk, upper Grande Ronde R, Lostine R
		Imnaha R, Little Sheep, Cow, Lightning Crks
		NF Clearwater R, Fish Crk, Selway R, Lochsa R
		SFSR & tribs, Big & Marsh Cks, Pahsimeroi, Salmon

Section 3. Focal species

primary: Chinook Snake River Spring/Summer ESU
secondary: Steelhead Snake River ESU
secondary: Westslope Cutthroat
secondary: Bull Trout

Section 4. Past accomplishments

Year	Accomplishments
2005	Cryopreserved gamete samples from 169 Chinook salmon representing 13 populations and 35 steelhead representing 3 populations. Conducted fertility trials, genetic and demographic analyses. 2004 Annual Report submitted. Management plan drafted.
2004	Cryopreserved gamete samples from 252 Chinook salmon representing 11 populations and 128 steelhead representing 3 populations. Genetic and demographic analyses. 2003 Annual Report submitted. Presented at the Aquaculture America meeting.
2003	Cryopreserved gamete samples from 266 Chinook salmon representing 12 populations and 90 steelhead representing 4 populations. Conducted fertility trials, genetic and demographic analyses. 2002 Annual report was submitted.
2002	Cryopreserved gamete samples from 286 Chinook salmon representing 12 populations and 283 steelhead representing 5 populations. Conducted fertiltiy trail, demographic and genetic analyses. 2001 Annual report submitted.
2001	Cryopreserved gamete samples from 398 Chinook salmon representing 12 populations and 295 steelhead representing 7 populations. Conducted fertility trials, genetic and demographic analyses. 2000 Annual Report submitted.
2000	Cryopreserved gamete samples from 349 Chinook salmon representing 11 populations and 283 steelhead representing 7 populations. Conducted fertility trials and genetic and demographic analyses. 1999 Annual Report submitted.
1999	Cryopreserved gamete samples from 340 Chinook salmon representing 8 populations and 212 steelhead representing 5 populations. Conducted fertility trials and genetic and demographic analyses. 1998 Annual Report submitted.
1998	Cryopreserved gamete samples from 295 Chinook salmon representing 10 populations and 25 steelhead representing 1 population. Genetic and demographic analyses. 1997 Annual Report submitted.

Section 5. Relationships to other projects

Funding source	Related ID	Related title	Relationship
BPA	199604300	Johnson Creek Artificial Propa	Collect and cryogenically store gamete samples from male Chinook salmon for use as germplasm source in the ongoing supplementation program.
BPA	199801001	Grande Ronde Captive Brood O&M	Provide a redundant storage facility for cryopreserved samples.
Other: Lower Snake River Compensation Program	[no entry]	NPT - Lower Snake River Compensation Program	LSRCP provides cost sharing by providing collection assistance.
BPA	199107200	Redfish Lake Sockeye Salmon Ca	Provide storage facility for cryopreserved gamete samples
BPA	198806400	Kootenai R White Sturgeon	Provide long term storage facility for cryopreserved sturgeon gametes (sperm).
BPA	199009300	Genetic Analyses of Oncorhynch	Provide DNA from multiple populations for analysis

Section 6. Biological objectives

Biological objectives	Full description	Associated subbasin plan	Strategy
Analysis of the diversity in the gene bank	An important objective of the Salmonid Gamete Preservation project is to determine the diversity of the fish that contributed gametes to the gene bank and compare it to that of the source populations. Diversity will be evaluated using genetic and demographic parameters and results will determine when adequate diversity has been preserved relative to the parameters outlined in the collection strategies presented above. Within and among-population measures of genetic differentiation will be assessed using Fst, heterozygosities and allelic diversity. This information will evaluation of the genetic diversity contained in the gamete repository, help determine genetic structure of Chinook salmon and steelhead populations and serve as a baseline to monitor shifts or losses of genetic variation over time. Chinook salmon genotyping will be done at Hagerman National Lab using the standardized set of 13 microsatellite loci. Steelhead genotyping will be done at a lab to be determined using the standardized set of microsatellite loci. Demographic data analysis includes determining the age structure of males that contributed gametes to the repository and comparing it to the source populations by year. In addition, effective brood year analysis (Young and Kucera, 2004) will be performed to approximate the brood year contribution of donor males. Briefly, this analysis involves determining the brood year for each donor, then looking at the distribution of donors over multiple generations. This analysis can identify underrepresented brood years and enable targeted collections of fish from specific brood years. Evenly collecting across brood years preserves the greatest level of diversity. Results will be communicated in Annual Reports and peer reviewed publications.	Salmon	Objective 2A: By 4th field hydrologic unit, carry out focused activities designed to improve our understanding and definition of small populations, while protecting the genetic integrity of wild populations that are below historical levels.
Coordinate gamete preservation among agencies	Salmonid gamete preservation and related activities will be coordinated with co-managers in the Snake River basin. The Nez Perce Tribe strongly believes in coordination efforts to monitor, conserve, protect and recover populations at low levels of abundance and high risk of extirpation. Coordinatin with other agencies and Tribes will assist with the collection of gametes and promote the wise use of the cryopreserved samples to accomplish recovery objectives.	Salmon	66A5. Coordinate plan implementation with federal, tribal, state, local, and other interests and avoid program and project duplication. 66A9. Provide opportunities for subbasinwide information distribution...
Cryopreservation research	Investigate novel methods of collecting, handling and cryogenic freezing and thawing with annual experiments. This will not only directly benefit this project, but will also benefit other BPA/NPPCC supported projects in the basin by potentially improving the cryotechnology and increasing the fertility rate of cryopreserved sperm. Currently cryogenic technology is limited by the relatively low survival of fish produced using cryopreserved gametes. This objective will allow an adaptive management design that includes experimentation and measurable results that will determine address scientific uncertainties associated with cryogenic technology. Increasing this survival would increase the usefulness of the technology and promote its application in other programs. Future experiments addressing key uncertainties associated with cryogenic technology will be developed based on project evaluation and communication with other gene conservation projects. Results will be communicated in Annual Reports and peer reviewed publications. Planned future research includes testing new cryoextenders that have the potential to increase the fertility rate of cryopreserved sperm (2006), investigating field cryopreservation methods (2006), understanding the number of eggs that can be fertilized using different volumes of cryopreserved milt (2007), investigating novel methods of freezing larger volumes of milt (5.0 ml; 2007) and further evaluating the viability of fish produced using cryopreserved sperm (2008). All research will involve testing the hypothesis that the treatment group is not significantly different from the control group.	Salmon	1B3. Implement innovative propagation techniques to meet goals identified in Table 6 for anadromous salmonids.
Preserve Genetic diversity in the Clearwater subba	Gamete collection and preservation from ESA listed Snake River ESU steelhead in the Clearwater subbasin will continue from Fish Creek, Selway River, Northfork Clearwater River and the upper Lochsa and tributaries. Gametes will be collected from other locations in the subbasin if the opportunity presents itself (additional wiers or traps are constructed). Ongoing collections will continue from major spawning aggregations until the collection strategy goals are met (see narrative section F).	Clearwater	D1: Maximize hatchery effectiveness in the subbasin. D2: Strategy: Apply safety net hatchery intervention based on extinction risk analysis and benefit risk assessments.
Preserve genetic diversity in the Grande Ronde sub	Gamete collection and preservation from ESA listed Snake River Sp/Su Chinook salmon and steelhead in the Grande Ronde River subbasin will continue. Chinook salmon gametes will be collected from upper Grande Ronde River, Catherine Creek and Lostine River fish spawned at Lookingglass Hatchery. Steelhead gametes will be collected from fish spawned or captured in traps or wiers in the subbasin. Gametes will be collected from other locations in the subbasin if the opportunity presents itself (additional wiers or traps are constructed). Ongoing collections will continue from major spawning aggregations until the collection strategy goals are met (see narrative section F).	Grande Ronde	Management Objective 3: Assess life history characteristics and maintain genetic diversity in supplemented and unsupplemented focal populations in the Grande Ronde
Preserve genetic diversity in the Imnaha subbasin	Gamete collection and preservation from ESA listed Snake River Sp/Su Chinook salmon and steelhead in the Imnaha River subbasin will continue. Chinook salmon gametes will be collected from Imnaha River fish spawned at Lookingglass Hatchery and steelhead gametes will be collected from Little Sheep Creek trap and wiers on Cow and Lightning Creeks. Gametes will be collected from other locations in the subbasin if the opportunity presents itself (additional weirs or traps are constructed). Ongoing collections will continue from major spawning aggregations until the collection strategy goals are met (see narrative section F).	Imnaha	Strategy 3A2: Continue ongoing programs: In areas were intervention has already occurred, support refinement of genetic preservation techniques such as captive brood stock, cryopreservation and supplementation.
Preserve genetic diversity in the Salmon River sub	Gamete collection and preservation from ESA listed Snake River Sp/Su Chinook salmon and steelhead in the Salmon River subbasin will continue. Chinook salmon gametes will be collected from SFSR, Lake Creek, Johnson Creek, Big Creek, Marsh Creek, Capehorn Creek, Pahsimeroi River, upper Salmon River and other locations in the basin. Steelhead gametes will be collected from fish spawned or captured in the SFSR, Johnson Creek, Pahsimeroi River and other locations in the basin. Gametes will be collected from other locations in the subbasin if the opportunity presents itself (additional wiers or traps are constructed). Ongoing collections will continue from major spawning aggregations until the collection strategy goals are met (see narrative section F).	Salmon	[Strategy - 2A2. Continue ongoing and develop new programs in areas where intervention has already occurred and in order to meet interim abundance and delisting targets. Support refinement of genetic preseration techniques... cryopreservation.
Preserve genetic diversity in the Snake R. basin	Columbia River salmonid populations have experienced significant decline in population numbers over the past five decades with Snake River Chinook salmon and steelhead now listed as "threatened" under the Endangered Species Act (ESA). In spite of the recent large returns of Chinook salmon and steelhead from 2001 - 2004, the return of anadromous salmonids in 2005 was low, and projected to be lower still in 2006. From this it is clear that recovery through habitat protection, population protection and harvest monitoring is not certain. Given this uncertainty, the establishment of a gene bank to preserve the genetic diversity of the remaining salmonid populations in the Snake River basin is warrented. To do this we propose to continue collecting and cryogenically preserving gametes (sperm) from populations of Chinook salmon, steelhead and bulltroutat in the region. These efforts are needed to preserve and maintain salmonid population genetic diversity, as an insurance policy against population collapse and extirpation, for ongoing artificial propagation programs and to preserve genetic material for future management options. Genetic conservation using gamete cryopreservation was a major objective of the Salmon, Imnaha and Grande Ronde River subbasin plans. Similar strategies were an important part of the Clearwater, Imnaha and Grande Ronde subbasin plans. The structured formatt made it impossible to include them in this proposal form.	Salmon	2A1. Preserve the genetic integrity of existing wild stocks in the Salmon Subbasin. Apply gene conservation measures (cryopreservation) to prevent irretrievable loss of genetic diversity. 2A2. Support the refinement of genetic preservation techniques

Section 7. Work elements (coming back to this)

Work element name	Work element title	Description	Start date	End date	Est budget
Produce Environmental Compliance Documentation	ESA compliance and documentation	Submit required documents to BPA, NOAA Fisheries and USFWS to ensure NEPA/ESA compliance is met for sampling ESA-listed species and working in regions inhabited by chinook salmon, steelhead and bulltrout.	1/1/2007	12/31/2007	$9,000
Biological objectives Coordinate gamete preservation among agencies			Metrics
Other	Chinook Salmon and Steelhead Gamete Collection	Significant numbers of gamete samples have been preserved from many populations in the Snake River basin. However, collections from many significant wild populations remain small, and even many of the collections from hatchery populations consist largely of hatchery-origin fish. Continuing this project will increase the number of preserved gamete samples from the imperiled wild populations and allow the collection of gametes from additional natural-origin fish from hatchery-integrated systems. Hatchery- and natural-origin fish interaction in an integrated hatchery program adds a additional threat genetic diversity persistence and increases the need to preserve the remaining natural origin diversity from a population. Gametes from adult male chinook were collected at adult traps, hatcheries or netted from streams. Collection from hatcheries or adult traps is coordinated with hatchery managers and done in cooperation with the hatchery staff. Generally the NPT gamete collection crews arrived at the hatchery or trap on spawning days and collected gametes from previously spawned or excess fish. If available, we also collected gametes from wild or hatchery fish in the trap prior to their being released upstream to spawn naturally. This was the most effective method of collecting gametes from a population and allowed us to preserve gametes from large numbers of fish. Collection of gametes from naturally spawning males is more labor intensive and not nearly as effective as collecting from hatcheries and traps. However, gamete collection from naturally spawning fish is extremely important to the project as it enables the preservation of genetically diverse populations. The NPT collect crews relied on stream survey data from ongoing monitoring and evaluation projects to locate areas of actively spawning salmon. Crews of four to seven people, including one or two in dry suits and snorkeling gear, walk the stream looking for lone males well separated from actively spawning fish. When an actively spawning female or male/female pair is identified, personnel are directed to walk around at a distance that did not spook the fish. In addition, collection crews are directed to walk on the stream bank and cross the stream in deep pools or shallow riffles in order to prevent trampling on redds. These measures minimize the impact on natural spawning activity. When a male is located, personnel set large dip nets across a narrow location in the steam below the fish. A snorkeler enters the water above the fish and attempts to guide the fish into the nets. Fish often swam under an undercut bank requiring the snorkeler to locate and capture the fish by grasping it around the caudal peduncle, slowly pull it out and guide it into a dip net. Nearly dead males are occasionally located and captured by hand. Following capture the fish is held in the water until a 95 liter portable anesthesia tank is set up and filled with 100 mg/l tricane methanesulfonate (MS-222, FinquelTM) with 200 mg/l of sodium bicarbonate buffer (NaHCO3; Allen and Harman, 1970). The fish is placed into the anesthesia tank until it loses equilibrium, generally 10 to 90 seconds. It is then rinsed in fresh water, the vent area is dried and milt is stripped into a whirlpac ™ bag. Generally 5.0 ml of milt is collected from Chinook salmon. The bag containing the milt is immediately inflated with air using a small foot pump and placed in a cooler containing ice. The bag is insulated from the ice by two layers of newspaper to prevent it from becoming too cold. The fish is placed in a reservoir of fresh water on top of the tank to begin recovery while general biological data such as fork length, mid-eye to hypural plate length, general condition and external marks are recorded. Tissue is collected and preserved in 95% non-denatured ethanol for later genetic (DNA) analysis and scales are taken for age assessment using scale pattern analysis. Finally, the fish is placed in the stream and revived by hand until it swims away. Gametes from adult steelhead are collected at adult traps or by angling. Collection at adult traps is similar to that described above for Chinook salmon. Naturally spawning males are captured by angling using artificial lures and barbless hooks. High water in the spring combined with the fact that steelhead often spawn in larger rivers makes net capture unfeasible. Angler hooked fish are brought in as rapidly as possible, netted and sex determined without removing the fish from the water. Females are immediately released. Males are handled similar to that for Chinook salmon described above.	3/1/2007	10/1/2007	$352,280
Biological objectives Preserve Genetic diversity in the Clearwater subba Preserve genetic diversity in the Grande Ronde sub Preserve genetic diversity in the Imnaha subbasin Preserve genetic diversity in the Salmon River sub Preserve genetic diversity in the Snake R. basin			Metrics
Other	Cryogenically preserve male gametes	Utilize cryogenic technology in the long term preservation of male gametes. A majority of this work is performed by WSU and UI and administered under subcontracts. If volume was adequate samples from each male were equally divided into two bags and cryopreserved at both universities. Milt was frozen in a total of 40 0.5ml French straws (IMV International, Minneapolis, MN), 20 by each university. Steelhead generally only produced 1.0 to 2.0 ml of milt so samples were generally preserved using 20 0.5 ml French straws. From 1992 through 2002 surplus milt was frozen in 5.0 ml straws (Wheeler and Thorgaard, 1991) however, we recently stopped using 5.0 ml straws due to problems with the thawing process. Each sample was given a unique straw code that could be cross referenced to a database containing collection information. Sperm quality was determined by estimating the percentage of motile sperm following the addition of a sperm activating solution.	3/1/2007	10/1/2007	$177,000
Biological objectives Preserve Genetic diversity in the Clearwater subba Preserve genetic diversity in the Grande Ronde sub Preserve genetic diversity in the Imnaha subbasin Preserve genetic diversity in the Salmon River sub Preserve genetic diversity in the Snake R. basin			Metrics
Other	Gamete transport	Transport 300 chinook salmon and 200 steelhead gamete samples from hatcheries or remote field locations to WSU or UI where they are cryogenically frozen within 24 hours. Gametes are transported using fixed wing aircraft or a vehicle to WSU/UI. Aircraft transport is performed by a subcontractor.	3/1/2007	10/1/2007	$77,000
Biological objectives Preserve Genetic diversity in the Clearwater subba Preserve genetic diversity in the Grande Ronde sub Preserve genetic diversity in the Imnaha subbasin Preserve genetic diversity in the Salmon River sub Preserve genetic diversity in the Snake R. basin			Metrics
Other	Maintain gene bank inventory	For security reasons, a largely duplicate gene bank is house in two locations at Washington State University and University of Idaho. This work element is performed by subcontractors at each location and administered through subcontracts. Samples are stored in large MVE (MVE International Corp., Maple Grove, MN, USA) or Taylor-Wharton (Taylor Wharton Cryogenics, Theodore, AL, USA) cryogenic freezers supplied with weekly deliveries of liquid nitrogen using 180 L cylinders. Currently gamete samples from 2492 chinook salmon, 1336 steelhead, 15 Wenachee River coho salmon, 22 Kotennei River sturgeon and 9 Kotennei River burbot are preserved in the gene bank. In addition, gamete samples from the Lostine River, Catherine Creek and Grande Ronde River captive brood programs are held in the genebank. The tanks are monitored and have an alarm system that alerts when the nitrogen level falls below a preset level. When this occurs the tank still contains enough nitrogen to maintain the samples for at least a week, but nitrogen should be added as soon as possible.	1/1/2007	12/31/2007	$73,000
Biological objectives Preserve Genetic diversity in the Clearwater subba Preserve genetic diversity in the Grande Ronde sub Preserve genetic diversity in the Imnaha subbasin Preserve genetic diversity in the Salmon River sub Preserve genetic diversity in the Snake R. basin			Metrics
Coordination	Coordination with regional and federal management agencies	This project will be coordinated with all ongoing Lower Snake River Compensation Plan (LSRCP), Idaho Power Company, ODFW, IDFG, WDFW, NOAA Fisheries, USFWS, other NPT projects and captive broodstock artificial propagation programs that utilize salmonids for recovery efforts. Coordination with fisheries management agencies and Tribes will occur prior to the implementation of this project. Also coordinate with the WSU, UI and NOAA Fisheries Center for Reproductive Biology, Salmonid Recovery group. This group works on reproductive aspects related to the recovery of salmonid populations in the Pacific Northwest. Based out of WSU, the center provides scientific support for this project by its world renowned faculty and facilities. Coordinate with the USDA National Animal Germplasm Program, Aquatic Species Committee (NAGP) based in Fort Collins, CO. This agency manages the national animal species genebank and is funded to preserve genetic diversity of agriculturally important animal species. The Aquatic Species Committee meets regularly to provide assistance with all aspects of developing and managing an aquatic species gene bank.	1/1/2007	12/31/2007	$50,000
Biological objectives Coordinate gamete preservation among agencies			Metrics
Manage and Administer Projects	Subcontract Administration	Four main subcontractors assist the completion of this project. They include 1)WSU and UI for performing the cryopreservation of sperm samples, maintaining the gene bank inventory and housing and maintaining the storage tanks ($30,000/year for each university); 2) A/L Compressed Gases (Spokane, WA) for providing weekly delivery of liquid nitrogen and other cryotechnology equipment and services ($26,000/year) and; 3) Interstate Aviation (Pullman, WA) for providing air transport of samples from the collection sites to the universities ($10,000 - $12,000/year). All subcontractors were low bidders for their services and are under contract with the Nez Perce Tribe. This WE also includes dealing with contractual obligations of BPA (SOW and budget submission), BPA training and NPT training.	1/1/2007	12/31/2007	$83,000
Biological objectives Coordinate gamete preservation among agencies			Metrics
Produce Annual Report	Annual Report	Produce annual report summarizing work accomplished in the previous contract period.	1/1/2007	3/1/2007	$30,000
Biological objectives Coordinate gamete preservation among agencies			Metrics
Produce Status Report	Quarterly reports	Write quarterly progress report detailing significant events, both positive and negative, that occurred in the previous quarter.	4/1/2007	12/31/2007	$6,000
Biological objectives Coordinate gamete preservation among agencies			Metrics
Analyze/Interpret Data	Analyze/Interpret data	An important objective of the Salmonid Gamete Preservation project is to determine the diversity of the fish that contributed gametes to the gene bank and compare it to that of the source populations. Diversity will be evaluated using genetic and demographic parameters and results will determine when adequate diversity has been preserved relative to the parameters outlined in the collection strategies presented above. Within and among-population measures of genetic differentiation will be assessed using Fst, heterozygosities and allelic diversity. This information will evaluation of the genetic diversity contained in the gamete repository, help determine genetic structure of Chinook salmon and steelhead populations and serve as a baseline to monitor shifts or losses of genetic variation over time. Chinook salmon genotyping will be done at Hagerman National Lab using the standardized set of 13 microsatellite loci. Steelhead genotyping will be done at a lab to be determined using the standardized set of microsatellite loci. Demographic data analysis includes determining the age structure of males that contributed gametes to the repository and comparing it to the source populations by year. In addition, effective brood year analysis (Young and Kucera, 2004) will be performed to approximate the brood year contribution of donor males. Briefly, this analysis involves determining the brood year for each donor, then looking at the distribution of donors over multiple generations. This analysis can identify underrepresented brood years and enable targeted collections of fish from specific brood years. Evenly collecting across brood years preserves the greatest level of diversity. Results will be communicated in Annual Reports and peer reviewed publications.	1/1/2007	12/31/2007	$81,000
Biological objectives Analysis of the diversity in the gene bank			Metrics Primary R, M, and E Type: Fst, heterozygosity, effective population size
Collect/Generate/Validate Field and Lab Data	Collect and generate field data	Biological data to be collected includes fork length, mideye – hypural length, scales (for aging), tags or marks and tissue samples (for DNA analysis). These data will assist in the evaluation of this project and provide important data for regional agencies. Results will be communicated in Annual Reports and peer reviewed publications.	3/1/2007	10/1/2007	$29,000
Biological objectives Analysis of the diversity in the gene bank			Metrics Primary R, M, and E Type: demographic data of fish contributing gametes
Collect/Generate/Validate Field and Lab Data	Collect and generate lab data	Currently cryogenic technology is limited by the relatively low survival of fish produced using cryopreserved gametes. This objective will allow an adaptive management design that includes experimentation and measurable results that will determine address scientific uncertainties associated with cryogenic technology. Increasing the fertility of cryopreserved gametes would increase the usefulness of the technology and its application in other programs. Future experiments addressing key uncertainties associated with cryogenic technology will be developed based on project evaluation and communication with other gene conservation projects. Results will be communicated in Annual Reports and peer reviewed publications. Planned future research includes testing new cryoextenders that have the potential to increase the fertility rate of cryopreserved sperm (2006), investigating field cryopreservation methods (2006), understanding the number of eggs that can be fertilized using different volumes of cryopreserved milt (2007), investigating novel methods of freezing larger volumes of milt (5.0 ml; 2007) and further evaluating the viability of fish produced using cryopreserved sperm (2008). All research will involve testing the hypothesis that the treatment group is not significantly different from the control group.	1/1/2007	12/31/2007	$77,000
Biological objectives Cryopreservation research			Metrics Primary R, M, and E Type: Percent fertility using cryopreserved sperm
Disseminate Raw/Summary Data and Results	Conference attendance and presentations	Attend conferences and disseminate data. Present significant project results at a regional and/or national conference including Idaho AFS, National AFS, American Aquaculture Society and other regional or national conferences. Attend regional workshops. Each person under the contract will attend no more than one conference.	1/1/2007	12/31/2007	$12,000
Biological objectives Coordinate gamete preservation among agencies			Metrics

Section 8. Budgets

Itemized estimated budget

Item	Note	FY07	FY08	FY09
Personnel	Project Leader, 1/2 Tech II, seasonal Tech I's, admin support	$114,938	$120,685	$126,719
Fringe Benefits	Project Leader, 1/2 Tech II, seasonal Tech I's, admin support	$31,777	$33,365	$35,033
Travel	Field per dium, coordination travel	$5,340	$5,340	$5,340
Supplies	field, lab and office supplies, computer upgrade in 2008	$5,090	$8,090	$5,090
Overhead	indirect rate, phone, vehicle, rent, utilities	$68,680	$72,342	$74,351
Other	subcontracts; nitrogen, gamete transport flights, university cryo and maintainance	$113,700	$114,700	$115,700
Totals		$339,525	$354,522	$362,233

Total estimated FY 2007-2009 budgets

Total itemized budget:	$1,056,280
Total work element budget:	$1,056,280

Cost sharing

Funding source/org	Item or service provided	FY 07 est value ($)	FY 08 est value ($)	FY 09 est value ($)	Cash or in-kind?	Status
Lower Snake River Compensation Program	Personnel, vehicles	$12,523	$13,149	$13,806	In-Kind	Confirmed
University of Idaho	Consultation services, Dr. Joseph Cloud	$4,000	$4,000	$4,000	In-Kind	Confirmed
Totals		$16,523	$17,149	$17,806

Section 9. Project future

FY 2010 estimated budget: $362,207
FY 2011 estimated budget: $362,207

Comments: [Outyear comment field left blank]

Future O&M costs: The salmonid gene bank housed at the University of Idaho and Washington State University is the product of over 10 years of collections, incuding BPA-supported collections since 1997, and over a million dollars of direct investment. If the current proposal is not funded, a contingency plan must be developed that will preserve the highly valuable samples. There are two options for these samples. First, the samples in the gene bank require constant maintainance and will quickly degrade if the liquid nitrogen supply is not continued. At a minimum, this will require approximately $30,000 per year in nitrogen, plus associated maintainance costs of renting floor space and personnel required to check the samples (currently it costs $18,000 for this service with the floor space rental provided by the universities). Second, the samples can be transfered to the United States Department of Agriculture, National Animal Germplasm Repository in Fort Collins, CO for long term storage. The storage is free, but it will cost approximately $50,000 to transfer the samples to the facility.

Termination date: none
Comments: This project will continue until adequate diversity is preserved and recovery of threatened salmonids in the Snake River basin is certain.

Final deliverables: A gene bank containing viable sperm samples representing the diversity of Chinook salmon, steelhead and other threatened species of the Snake River basin.

Section 10. Narrative and other documents

Reviews and recommendations

FY07 budget	FY08 budget	FY09 budget	Total budget	Type	Category	Recommendation
NPCC FINAL FUNDING RECOMMENDATIONS (Oct 23, 2006) [full Council recs]
$65,000	$65,000	$65,000	$195,000	Expense	Multi-province	Fund
NPCC DRAFT FUNDING RECOMMENDATIONS (Sep 15, 2006) [full Council recs]
$0	$0	$0	$0		ProvinceExpense
NPCC DRAFT FUNDING RECOMMENDATIONS (Sep 15, 2006) [full Council recs]
$65,000	$65,000	$65,000	$0		Multi-province
Comments: Reduced or removed due to ISRP concerns. MSRT recommends 308,447.

ISRP PRELIMINARY REVIEW (Jun 2, 2006)

Recommendation: Fundable (Qualified)

NPCC comments: Previous comments from the Provincial/Systemwide Review still apply. To repeat, "It is time for a thoughtful analysis of what the Fish and Wildlife Program wants to accomplish in gene conservation, whether or not cryopreservation continues to be a useful tool, and whether an ever-increasing commitment to this program is consistent with that goal." The project applies to a number of subbasins and to ESA planning in the Columbia River Basin, and should benefit focal species. The program could turn out to be critically important if ecosystem dysfunction issues can be addressed and "old" genes are wanted to expand the diversity in the extant population. The proposal was clearly written and provided a strong technical and scientific background. There was evidence of excellent collaboration between the project and agencies, tribes, and universities. The project history is well described. Proponents and cooperators publish in the peer-reviewed literature and report at regional and national conferences. The narrow, task-oriented objective that was provided should have been superceded by an overarching objective dealing with preserving genetic diversity in ESA-listed fishes. Reviewers would have appreciated more evidence that project personnel are able to keep up with all emerging technologies. Another important issue, not apparently addressed in the proposal, is when is active collection of sperm completed, and thus when is it appropriate for the proposal to shift into a lower budget maintenance mode. Also, if future monitoring indicates a significant reduction in sperm viability then there would be a need to rotate or update earlier collections. This should be addressed at some point in time.

ISRP FINAL REVIEW (Aug 31, 2006)

Recommendation: Fundable (Qualified)