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 Bacterial Kidney Disease Aeromonas salmonicida Furunculosis *Will see other environmental, non-pathogenic bacteria Virus Infectious Hematopoietic Necrosis Virus (IHNV)

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

Primary capture methods E-fishing Raft Snorkel Seining Backpack Shocker

Mapping Sites

Labeling Fish for Transport

Pathogen Diagnostics Full Necropsy Bacteriology Virology Parasite Identification

Final Numbers Total 299 Salmonids 242 -Above 81 -Below 161

Unknown Cestode Rhabdochona Unknown Trematode

Apophallus sp. Myxobolus squamalis Nanophyetus Myxobolus insidiosus

Copepod

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

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

Pathogen Detection Water Sampling Sascha Hallett Pathogen Detection Water Sampling

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

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

Water Sampling

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

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

Questions?