Salmonid Pathogens in the Columbia Cascade Province

Frank Loge1 Douglas Call2

Michael Barber1

1Department of Civil and Environmental Engineering

2Department of Veterinary Microbiology & Pathology and WSU/UofI Center for

Reproductive BiologyWashington State University

Salmonid pathogen program

Immunopathology•Diagnostics•Surveillance•Vaccine development

Ecology•Environmental sampling•Risk-based analysis•Epidemiology

Genomics & proteomics•Marker identification•Differential virulence•Phylogeny

Hypotheses

1. Source affects pathogen load

2. Anthropogenic pollution affects in-stream survival and propagation of pathogens

3. Pathogen concentrations vary with light, temperature and aquatic habitat

Rationale

• 14 salmonid species are listed as either threatened or threatened.

• Disease impacts every stage of salmonid life-history.

• Very little information available on ecology of infectious disease agents relative to fish populations, watersheds or habitats.

Significance• Factors that contribute to large-scale

salmonid infections are unknown and represent a critical data gap (Upper Middle Mainstem Subbasin report)

• Consistent with Northwest Power Planning Council Fish and Wildlife Program

• Consistent with NMFS and USFWS 2000 Biological Opinion and Action Agencies Implementation Plan (actions 141, 184, 11.2, 9.6.1.1)

Objectives• Develop a microarray detector and

sampling protocol for multiplex detection of salmonid pathogens

• Characterize the spatial and temporal occurrence of pathogens and water quality parameters

• Identify possible sources of salmonid pathogens and correlate with anthropogenic pollution

Microarrays permit efficient multiplex analysis with multiple

internal controlsCP1 CP2R.s. A.s. Y.r. A.s. Y.r. R.s.

p57F p57RProbe A Probe B

16S rDNA

p57 gene

Why use microarrays?

Detecting BKD (Renibacterium salmoninarum)

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Overview of monitoring protocol

Collect water sample

Conventional water quality

analysis

Filtration

Extract RNA

Extract DNA

Make cDNA

Multiplex PCR

Microarray detection

Sampling strategyFish Pathogens

Flow

Point sourcedischarges

Non-pointsourcerunoff

Spatial and temporalvariations influenced by

Light Temperature Habitat

Expected Results and Benefits

• Provide a field deployable monitoring protocol and a risk-based methodology for interpreting results

• Provide critical information on the temporal and spatial occurrence of planktonic salmonid pathogens

• Identify possible sources, fate, and transport of salmonid pathogens and relationships with specific water quality characteristics