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

2. Specific Pathogens of Concern
Bacteria
Flavobacterium psychrophilum
Cold Water Disease
Flavobacterium columnare
Columnaris Disease
Renibacterium salmoninarum
Bacterial Kidney Disease
Aeromonas salmonicida
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
Salmonids 242
-Above 81
-Below 161

10. Unknown Cestode
Rhabdochona
Unknown Trematode

12. Apophallus sp.
Myxobolus squamalis
Nanophyetus
Myxobolus insidiosus

14. Copepod

16. 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

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

18. Pathogen Detection Water Sampling
Sascha Hallett
Pathogen Detection
Water Sampling

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

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

21. Water Sampling

22. Water Sampling Bacteria Extract total DNA Quantify using qPCR
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)
Unchartered territory
Incidental, nonpathogenic bacteria/organisms also present

23. 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
Use qPCR to enumeraet a given pathogen in a water sample

24. Questions?