1. Nez Perce Tribe Department of Fisheries Resources Management William Young, Paul Kucera and Jay Hesse The Nez Perce Tribe Listed Stock Salmonid Gamete Preservation

2. Snake River spring/summer chinook Snake River steelhead –Both listed as threatened under the ESA

3. Number of redds counted in 6 rivers in the Snake River basin from 1958-2002 Number of Redds Annual redd counts 10 year moving ave.

4. Project Chronology 1992 – First collections, LSRCP funded, UI and WSU collaborations 1996 – BPA proposal 1997 – First collections as a BPA funded project 2001 – Second BPA funding cycle

5. Regional Support and Collaborations Cost Share –LSRCP –WSU –UI Regional Support and Benefit –IDFG –WDFW –ODFW –Kootenai Tribe

6. Snake River Salmonid Gamete Preservation Plan Goal Preserve the genetic diversity of Pacific northwest salmonids for conservation and recovery efforts

8. Long term –Increase effective number of breeders - N b –Insurance against catastrophic population collapse –Maintain future options Short term –Conventional hatchery broodstock management Inadequate number of males Avoid sib matings –Captive broodstock programs Applications of Cryopreservation to Preserve Genetic Diversity

9. Collection Objectives 100 individuals / year for one generation from each spawning aggregation –Chinook Salmon –Steelhead

10. Effective Population Size - N e N e is the size of an ideal population that maintains genetic variation at the same rate as a real population (N) of interest Factors that cause a real population to diverge from an ideal population –unequal sex ratios –nondiscrete generations –nonrandom mating –unequal family sizes –variable population size

11. The relationship between N b and N N b /N ratios for anadromous fish are typically 0.1 to 0.4 N b of approximately 50 –95% probability of rare allele retention Requires N = 125-500 / generation

12. Total genetic diversity Among population genetic diversity

13. Marsh Creek Upper Salmon Imnaha River Lostine River Pahsimeroi River South Fork Salmon River Rapid River Big Creek Lake Creek UPGMA – Neis Genetic Distance Among Population Genetic Diversity of Cryopreserved Samples

14. Within Population Genetic Diversity Among brood year genetic variation

15. Effective Brood Year Analysis A way to measure the composition of gamete samples collected from adults spanning multiple generations. Strive for an even representation of fish from different brood years across multiple generations.

16. EBY 1EBY 2EBY 3EBY 4EBY 5 Year 1 Year 2 Year 3 Year 4 Year 5 5 3 4 5 3 4 5 3 4 5 4 3 5 3 4 Total 100

17. Gamete Collections Under Ideal Circumstances

18. EBY 1EBY 2EBY 3EBY 4EBY 5 Year 1 Year 2 Year 3 Year 4 Year 5 Year 6 Year 7 5 3 4 5 3 4 5 3 4 5 3 4 5 3 4 5 4 3 5 3 4. Year n Total 23 16 38 26 33 14 41

19. SFSR Chinook Salmon – Effective Brood Year analysis

20. Project Successes Largest salmonid genebank in North America 2240 chinook salmon gamete samples preserved from 13 spawning aggregations 1210 steelhead gamete samples preserved from 12 spawning aggregations Annual fertilization trials Annual genetic analyses Numerous experiments aimed at improving the viability of cryopreserved sperm Experiments aimed at assessing the viability of fish produced using cryopreserved sperm

21. Chinook Salmon Gamete Collections, 1992 - 2004

22. Chinook Salmon Gamete Collections, 1992 - 2003

23. TRT Chinook Salmon Populations Grande Ronde Basin –Catherine Creek –Wallowa/Lostine Rivers –Minam River –Upper Grande Ronde –Wenaha River Imnaha Basin –Big Sheep Creek –Imnaha mainstream Lower Snake Basin –Tucannon River –Asotin Creek South Fork Salmon Basin –Secesh River –SFSR –EFSFSR/Johnson Creek Salmon River Tribs –Chamberlain Creek –Little Salmon River Middle Fork Salmon Basin –Bear Valley/Elk Creeks –Big Creek –Camas Creek –MFSR below Indian Creek –Pistol Creek –Marsh Creek –Sulphur Creek –Loon Creek –MFSR above Indian Creek Upper Salmon Basin –Valley Creek –Lemhi River –NF Salmon River –Pahsimeroi River –EF Salmon River –Upper Salmon River –Yankee Fork

24. Steelhead Gamete Collections, 1993 - 2004

26. TRT Steelhead Populations Lower Snake Basin –Tucannon River –Asotin Creek Clearwater Basin –Lower Clearwater –South Fork –Lolo Creek –Lochsa River –Selway River Grande Ronde Basin –Lower Grande Ronde –Joseph Creek –Wallowa River –Upper Grande Ronde Salmon Basin –Little Salmon –South Fork –Secesh River –Chamberlain Creek –Big, Camas, Loon –Upper Middle Fork –North Fork –Lemhi River –Pahsimeroi River –East Fork –Upper Salmon Imnaha Basin –Imnaha River Hells Canyon –Hells Canyon

27. Genetic Analyses – Chinook Salmon Average Heterozygosity – 2 microsatellite loci –Imnaha River.59206450 –Lostine River.5424101 –Rapid River.53147217 –Lake Creek.4748109 –SFSR.57192349 –Big Creek.6160133 –Marsh Creek.5830 87 –Pahsimeroi River.5388185 –Upper Salmon River.5879293 HN Total Samples

28. Genetic Analyses - Steelhead Mitochondrial Haplotype Analysis – D-loop sequencing –Dworshak 296414 –Little Sheep Creek350484 –Pahsimeroi River210424 –Oxbow Hatchery307445 TotalAnalyzed# haplotypes

29. Future Directions Continue to collect at current level Work to increase regional support for the project Maintain a collaboration with the USDA National Animal Germplasm Program Establish a regional repository

30. Acknowledgments Lower Snake River Compensation Project Bonneville Power Administration University Collaborators Joe Cloud, Gary Thorgaard, Paul Wheeler Nez Perce Tribe Jay Hesse, Mike Blenden, Mary Edwards, Nym Johnnie, Robyn Armstrong, Neal Espinosa, Cameron Albee, Joe McCormack, the McCall field office