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Published byAgata Tomczyk Modified over 5 years ago
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Copepod
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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
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Acknowledgments US Army Corp of Engineers ODFW Research Crew
ODFW Fish Pathology Oregon State University Bartholomew Lab
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Pathogen Detection Water Sampling
Sascha Hallett Pathogen Detection Water Sampling
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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
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Water Sampling 0.22µm Filter with vacuum pump – capture and concentrate pathogens
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Water Sampling
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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
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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
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Questions?
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