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Willamette River System

Willamette River System. 4 Hatchery Sites: Marion Forks South Santiam McKenzie – Leaburg Dexter Ponds. Specific Pathogens of Concern. Bacteria Flavobacterium psychrophilum Cold Water Disease Flavobacterium columnare Columnaris Disease Renibacterium salmoninarum

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Willamette River System

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  1. Willamette River System 4 Hatchery Sites: Marion Forks South Santiam McKenzie – Leaburg Dexter Ponds

  2. Specific Pathogens of Concern • Bacteria • Flavobacteriumpsychrophilum • Cold Water Disease • Flavobacteriumcolumnare • Columnaris Disease • Renibacteriumsalmoninarum • Bacterial Kidney Disease • Aeromonassalmonicida • Furunculosis *Will see other environmental, non-pathogenic bacteria • Virus • Infectious Hematopoietic Necrosis Virus (IHNV)

  3. Sampling Naturally Reared Fish • Capture naturally reared fish near hatchery • 2 efforts at each site targeting juvenile salmonids • July – August, based on ODFW availability • Additional above specimens collected from ODFW gillnet projects and screw traps • Fish euthanized and transported to OSU lab for diagnostics

  4. Primary capture methods E-fishing Raft Snorkel Seining Backpack Shocker

  5. Mapping Sites

  6. Labeling Fish for Transport

  7. Pathogen Diagnostics Full Necropsy • Bacteriology • Virology • Parasite Identification

  8. Final Numbers Total 299 Salmonids242 -Above 81 -Below 161

  9. Unknown Cestode Rhabdochona Unknown Trematode

  10. Apophallus sp. Myxobolussquamalis Nanophyetus Myxobolusinsidiosus

  11. Copepod

  12. Pathogens are not prevalent in these populations • Natural limitations, such as predation, make it difficult to sample epizootics in the field • Need to combine and analyze data from previous year • As monitoring continues: • Hope to collect fish if an outbreak occurs to see if any pathogens are detected • Need more samples above hatcheries for comparison • Pursue any environmental indicators of outbreaks in the hatcheries

  13. Acknowledgments • US Army Corp of Engineers • ODFW Research Crew • ODFW Fish Pathology • Oregon State University • Bartholomew Lab

  14. Sascha Hallett Pathogen Detection Water Sampling

  15. Water Sampling Bacteria - monitoring • 3 x 1L samples • influent, effluent • start and end of sentinel fish exposures = every week August through September Bacteria – outbreak • 3 x 1L samples • influent, afflicted ponds, effluent, 200 ft, 800 ft downstream • start and end of sentinel fish exposures

  16. Water Sampling 0.22µm Filter with vacuum pump – capture and concentrate pathogens

  17. Water Sampling

  18. Water Sampling Bacteria • Extract total DNA • dissolve filter paper in acetone • commercial kit • Quantify using qPCR • used for tissue samples but not water • limit of detection/sensitivity • specificity (especially in mixed DNA sample such as environmental water)

  19. Water Sampling Reference sample set • use qPCR to enumerate pathogen • translate qPCR output (Cq) • known quantities of pathogen • parallel dilution series of samples cultured and placed directly on filter paper

  20. Questions?

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