1. New genetic technology for the management of Columbia River salmon and steelhead Parentage Based Tagging Matthew Campbell Idaho Department of Fish and Game LSRCP PRODUCTION MEETING AGENDA MARCH 16 th, 2011 BOISE, IDAHO 1

2. “by tying a ribbon or thread to the tails of young salmon which were swimming toward salt water and catching them again when they came back to the same place upon their return from the sea” 2

3. In the Snake River basin, CWT recoveries are one of the primary tools used by managers in Oregon, Washington, and Idaho to: Estimate the number of hatchery salmon and steelhead contributing to fisheries in the Columbia River basin Estimate run-timing and harvest of individual hatchery stocks 3

4. Despite its’ predominance in addressing these management questions, CWT technology has several limitations: The process of physically tagging hundreds of thousands of juveniles from different hatchery stocks is logistically difficult labor intensive expensive potentially increases physiological stress 4

5. Parentage-based genetic tagging - PBT (Anderson and Garza 2005) Parentage-based tagging uses similar DNA techniques as those used in human parentage testing 5

6. Parentage-based genetic tagging - PBT Every person receives a unique set of genetic information from their parents! If your parents have been genetically screened they can easily be identified from thousands of possible candidates! 6 Child

7. Genetic information can be obtained from all hatchery fish (broodstock) that are spawned Sawtooth Pahsimeroi Oxbow Dworshak/Clearwater Lookinglass Wallowa 7

8. Offspring from any of these parents (either collected as juveniles or returning adults), if genotyped, could be assigned back to their parents, thus identifying the hatchery they originated from and exact brood year they were produced in Sawtooth BY2008 8

9. Any Snake River hatchery Chinook salmon or steelhead encountered in the basin (juvenile or adult) could be sampled, genotyped and its sex, exact age, and stock of origin could be determined 9 Snake River ESU

10. Benefits of PBT Provides same information as CWTs (stock and cohort) Run reconstruction (age, sex, stock of returning adults) Harvest estimates Additionally, many issues associated with tagging studies all but go away Tag loss Tag detectability Differential mortality 10

11. Additional benefits of PBT PROS: It is a passive mark (no handling of juveniles needed) “Tagged” fish can be non-lethally interrogated Fin-clip Scales Biopsy hook “The hollow stainless steel tip of the hook collects a small amount of tissue as the fish strikes, and a special rasp holds the sample in place while the hook is in the water” “Genetag hook” 11

12. Versatility of PBT PROS: Can potentially provide much more information than stock and cohort of origin Host of other life history, ecological and quantitative genetic questions 12

13. Versatility of PBT Hatchery reform (integrated programs) What is the reproductive success of hatchery-origin/wild- origin spawners? (both in the wild and in the hatchery) 13

14. Versatility of PBT Hatchery reform (segregated programs) Concern that straying hatchery fish reduce the fitness of wild populations Is it possible to manage a hatchery program so that it is truly “segregated” from wild populations? PBT technology can identify where straying hatchery fish were released, their stock of origin, and age 14

15. Versatility of PBT Heritability estimates To what extent are physical or behavioral traits (e.g. size and age of returning adults) controlled by genetics, environment or interactions between the two? Are hatchery spawning practices selecting for younger age at maturity in hatchery Chinook salmon populations 15 Hankin et al (2009) recommended that in the hatchery male length ≥ female length

16. Versatility of PBT Origin of kelt steelhead Thousands of kelt steelhead outmigrate past Lower Granite Dam each year (~50% hatchery origin) Where are they from? How old are they? Is phenotypic sex correct? 16

17. Versatility of PBT PBT program can easily be integrated into existing sampling/genetic programs at Bonneville Dam and in lower Columbia fisheries (CRITFC) Bonneville Chinook weekly stock composition (2004-2006) Collaborative Center for Applied Fish Science 17

18. Easily integrated into IDFG’s GSI program at LGR dam Genetic Stock Identification Versatility of PBT 18

19. PBT sounds great!!!.... However, it has been largely theoretical up to this point… Needs to be empirically tested and validated on a large scale 19 Snake River ESU

20. These types of evaluations have been directly called for by both the Pacific Salmon Commission and the Independent Scientific Review Panel and Advisory Boards 20

21. Snake River Chinook and Steelhead Parentage Based Tagging- Proposal #201003100 Perfect timing to conduct a large-scale test of this technology in the basin…. 21

22. Support from hatchery managers (ID, WA, OR): Sampled hatchery broodstock from all Snake River hatcheries (8 hatcheries, >45,000 samples in 2008, 2009 and 2010. (Both species-steelhead and spring/summer Chinook salmon) >45,000 samples !!! 22

23. Advancements in genetic technology: New genetic instruments and techniques allow more samples to be run at a quicker rate and for lower costs 96 samples in one run! 23

24. Strong collaboration among labs and the ability to standardize genetic marker sets: Allows multiple labs to process samples and share data Collaborative Center for Applied Fish Science Hagerman, ID Dr. Shawn Narum Dr. Jon Hess 24

25. Major objectives over the next 3 years: Continue with 100% PBT sampling at all Snake River hatcheries for spring/summer Chinook and steelhead. Complete transition from microsatellite DNA markers to “single nucleotide polymorphic” markers (SNPs) Identify a set of SNP markers that can achieve high parental assignment accuracy even when tens of thousands of potential parents are involved and when parentage involves hatchery populations that are very similar genetically due to shared ancestry or in situations where the broodstock contains closely related individuals Why? Low genotyping error rate Allele calls (nomenclature) are easily standardized between labs Amenable to high-throughput / low-cost genotyping 25

26. Total Columbia Snake River Releases Proportion Spring/Summer Chinook 24,000,00010,000,00042% Summer Steelhead 12,000,0008,000,00067% TOTALS 36,000,00018,000,00050% 26 This will effectively, permanently genetically “tag” ~18 million Snake River basin smolts, per year Major objectives over the next 3 years: Complete the first PBT broodstock baselines in the Columbia River basin

27. Major objectives over the next 3 years: We intend to demonstrate the applications of this technology through three sampling/genotype projects that will provide evaluations of both the accuracy and relative costs of PBT technology in providing equivalent CWT recovery data 27 1.Assessment of the stock composition and run timing of Snake River hatchery salmon and steelhead passing Bonneville Dam and harvested in mainstem Columbia River fisheries 2.An assessment of the origin of straying hatchery steelhead in the Salmon River basin 3.A paired CWT and PBT recovery experiment as part of existing LSRCP hatchery evaluations in Idaho

28. What progress have we made thus far on our objectives? 28

29. SNP development and testing In collaboration with CRITFC we identified 96 robust SNP markers (includes sex marker) Genetically diverse (all MAF >.20) Robust (easy to score, high genotyping rate)

30. SNP development and testing 96 SNP set is accurate!!! High number of juveniles assigned (97.3%) 100% accuracy to stock 100% accuracy to parents (error from datasheets not genetics) Hatchery Stock Juveniles Genotyped Juveniles Assigned % assigned correctly to stock # matching parents in cross records % matching parents in cross records Squaw Crk92 100.0% Sawtooth9391100.0%8593.4%* Dworshak93 100.0%93100.0% EF Salmon9488100.0% Pahsimeroi9388100.0% G.R. Cott.9391100.0%91100.0% Lyons Ferry930----

31. Broodstock genotyping Completed steelhead hatchery broodstock from 2008 (~5500 samples) Will have steelhead broodstock from 2009 and Chinook broodstocks from 2008 and 2009 completed by June 30 th Parent Database 31

32. Evaluations: Currently genotyping (N = 96) 1-0cean and (N = 96) 2-ocean CWT recovered adults (fall steelhead fisheries) CWT PBT Demonstrate accuracy of PBT in providing stock and age 1-ocean fish should assign, 2-ocean fish should not Results by the end of this month!!!!! 32

33. What’s up next? Sample and genotype kelts recovered at LGR this spring Complete similar “ground-truthing” exercises on CWT- recovered Chinook (2011/2012) Genotype all hatchery broodstock sampled in 2011 Continue to work with program managers on developing a strategy for transitioning from CWTs to PBT in the Snake River basin Continue to work with program managers on the development of the “rearing and release” module of the hatchery database. 33

34. Identify parents Stock and age Where it incubated Where it reared Where and when it was released Ultimate goal: Sawtooth Fish Hatchery Diet Disease Rearing density Temperature This information could be available for all hatchery broodstock every year! SEX 34 http://www.flickr.com/photos/natekay/4319654380/sizes/o/

35. Questions??? 35