genetic diversity of manayunkia speciosa in the klamath river basin by: dan horner mentors: sascha...
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Genetic diversity of Manayunkia speciosa
in the Klamath River basin
By: Dan HornerMentors: Sascha Hallett
Jerri Bartholomew
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Background
• Ceratomyxa shasta is a microscopic, spore-forming parasite
• Causes intestinal necrosis and mortality of Klamath River juvenile salmonids
• 60% of out migrating juvenile salmonids infected
• Severe economical and ecological impacts
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Economic Effects of C. shasta
• Low salmon numbers forced regulatory closures of 2006 salmon fishing
• Loss of $150 million to the region's economy
• Over $2 million spent on C. shasta research annually
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Salmonid Populations Salmonid species at risk:
- Spring and fall-run chinook
- Steelhead
- Coho salmon*- Coastal cutthroat trout
*Classified as an endangered species in 1997(ESA) and 2002 (CESA)
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C. shasta Lifecycle
MyxosporeActinospore
Salmonid
M. speciosa
Intraspecies differences could affect the suitability of this polychaete as the definitive host
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Hypothesis
• Manayunkia speciosa in the Pacific Northwest are the same species
• Determine if Klamath River polychaetes belong same species, and if so, determine if distinct strains of M. speciosa exist within the Klamath River Basin
Objective:
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Primary Locations of Research
Iron Gate Dam
Klamath Lake
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Willamette River
1. Acted as a necessary outgroup for intraspecific phylogenetic comparisons
2. A known infectious zone for C. shasta
3. 300 miles NW of primary sample site, the Klamath.
4. Genetically isolated river basin for comparison to the Klamath
Klamath
Willamette
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Manayunkia genetic background
• No genetic data exists for M. speciosa
• However, Manayunkia spp. have been sequenced from Lake Baikal
- M. sp.TP2002- M. zenkewitschii- M. athalasia
• All Manayunkia were sequenced using universal primers targeting the Cytochrome Oxidase c subunit 1 gene
Lake Baikal
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Methods
1. Extract DNA from collected samples
2. Specific CO1 gene amplified through PCR
3. PCR product sequenced and analyzed for novel genetic characteristics
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Cytochrome Oxidase Subunit 1(CO1)
Subunit 1
• CO1 gene:
- Could possibly be used as a universal barcode for all species
- Subunit 1 is a 710 bp mitochondrial gene
- Codes for 13 proteins
• For Nematodes:
- 10-20% variability: different species
- 0-6% variability: same species
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Species/Location Amplified # bp
d-valsp2002
d-valzenk
d-valathalasia
d-valWillamette
M. athalasia675 16% 22% - 19%
M. sp2002576
-19% 17% -
M. zenkewitschii565 20% - - -
Willamette520 - - 19% -
Williamson505 - - 19% 2.1%
I5 Site530 - - 19% 3.2%
Interspecies variance values consistent with known
species variability
Intraspecies variance also within acceptable level
Known species variability
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Current Genotypic Results
Klamath Genotypes
Willamette Genotypes
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Klamath and Willamette
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Conclusion
• All the Manayunkia used in previous C. shasta research have been the species, Manayunkia speciosa.
• Distinct genotypes appear to exist, but further research is necessary
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Ongoing Research
Fraser River, B.C.
Lake Superior, WI
Deschutes River, OR
Cowlitz River, WA
Hudson River, NY
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Acknowledgements• Stephen Atkinson• Gerri Buckles• Charlene Hurst• Sarah Bjork• Jill Pridgeon• Sue-Jie Koo• Kevin Ahern• Jerri Bartholomew• Sascha Hallett• HHMI