Pacific Oyster Physiological Response to
 Disease under Variable Environmental
                               Regimes

Emma...
Objective
    How do environmental stressors, such as ocean
     acidification (OA) and elevated temperature, affect
    ...
Experimental Plan
  How do environmental changes affect the host-pathogen relationship?


                                ...
Outline
    Ocean acidification
    Environmental stress and physiology
    Temperature and the C. gigas-V. tubiashii
 ...
Outline
    Ocean acidification
    Environmental stress and physiology
    Temperature and the C. gigas-V. tubiashii
 ...
Ocean Acidification
                                                     1000 ppm




     Increased CO2 in atmosphere


  ...
Environmental Stress
         Growth           Reproduction

                                    Normal Environment



Imm...
Outline
    Ocean acidification
    Environmental stress and physiology
    Temperature and the C. gigas-V. tubiashii
 ...
Temperature & V. tubiashii
    Two temperatures: 12oC & 25oC
    V. tubiashii challenge in both temperatures
    3 day ...
Temperature & V. tubiashii
                                     = 12oC
                                     = 25oC
       ...
Temperature & V. tubiashii

                       Assay Development




                 n=4
                            ...
Outline
    Ocean acidification
    Environmental stress and physiology
    Temperature and the C. gigas-V. tubiashii
 ...
Effects of OA on Development
    Strip-spawned and pooled sperm and eggs
    Fertilized eggs in pCO2-equilibrated water
 ...
Effects of OA on Development
                     Timeline of C. gigas Larval Development
Fertilization   2-cell
0h
       ...
Effects of OA on Development
=380 ppm
=840 ppm
           Proportion Eggs in Cleavage




                                 ...
Effects of OA on Development
=380 ppm
=840 ppm
       Proportion Larvae Hatched




                                   *
Effects of OA on Development
    A smaller proportion of larvae in elevated pCO2
     develop at the “normal” rate
    Im...
Effects of OA on Development
    General stress response genes
         Cytochrome P450 oxidase (CytP450)
         Perox...
Outline
    Ocean acidification
    Environmental stress and physiology
    Temperature and the C. gigas-V. tubiashii
 ...
OA & V. tubiashii
    Study in progress in collaboration with NWFSC (P.
     McElhany et al.)
    Planned 3 weeks: spawn...
= calcified
OA & V. tubiashii                                   = partially
                                              ...
Conclusions
    Ocean acidification has significant effects on C.
     gigas larval development.
    OA may inhibit and ...
Acknowledgements
    Funding: Saltonstall-Kennedy Grant (NOAA),
     NOAA Aquaculture Program student support
    Univer...
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Timmins-Schiffman P2010

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Timmins-Schiffman P2010

  1. 1. Pacific Oyster Physiological Response to Disease under Variable Environmental Regimes Emma Timmins-Schiffman, Elene Dorfmeier, Paul McElhany, Shallin Busch, Steven Roberts & Carolyn Friedman University of Washington PCSGA 2010
  2. 2. Objective   How do environmental stressors, such as ocean acidification (OA) and elevated temperature, affect oyster larvae survival and physiology when challenged by disease?
  3. 3. Experimental Plan How do environmental changes affect the host-pathogen relationship? High OA Temp. Host Pathogen Vibrio photo: Waldor & RayChaudhuri, 2000, Nature
  4. 4. Outline   Ocean acidification   Environmental stress and physiology   Temperature and the C. gigas-V. tubiashii relationship   OA effects on C. gigas   Current work: OA & V. tubiashii Hi OA T
  5. 5. Outline   Ocean acidification   Environmental stress and physiology   Temperature and the C. gigas-V. tubiashii relationship   OA effects on C. gigas   Current work: OA & V. tubiashii Hi OA T
  6. 6. Ocean Acidification 1000 ppm Increased CO2 in atmosphere 380 ppm Increased oceanic CO2 = lower oceanic pH
  7. 7. Environmental Stress Growth Reproduction Normal Environment Immune response General metabolism Environmental Stress Growth Reproduction Immune response General metabolism pH
  8. 8. Outline   Ocean acidification   Environmental stress and physiology   Temperature and the C. gigas-V. tubiashii relationship   OA effects on C. gigas   Current work: OA & V. tubiashii Hi OA T
  9. 9. Temperature & V. tubiashii   Two temperatures: 12oC & 25oC   V. tubiashii challenge in both temperatures   3 day trial   Immune response genes:   C-jun kinase   NfkB Up-regulate immune defenses: -ROS -inflammatory response
  10. 10. Temperature & V. tubiashii = 12oC = 25oC * =12oC + Vt * =25oC + Vt 1 2 3
  11. 11. Temperature & V. tubiashii Assay Development n=4 n=4 n=3 n=4 n=2 n=3 n=2 n=2 12oC 25oC 12oC 25oC 12oC 25oC 12oC 25oC
  12. 12. Outline   Ocean acidification   Environmental stress and physiology   Temperature and the C. gigas-V. tubiashii relationship   OA effects on C. gigas   Current work: OA & V. tubiashii Hi OA T
  13. 13. Effects of OA on Development   Strip-spawned and pooled sperm and eggs   Fertilized eggs in pCO2-equilibrated water   380 ppm   840 ppm   Time to developmental stage at   1 hour (2-cell)   2 hours (>4-cell)   5 hours (hatched)   17 hours   24 hours
  14. 14. Effects of OA on Development Timeline of C. gigas Larval Development Fertilization 2-cell 0h Developmental Stage 4-cell 1h hatching 2h 5h Time Post-Fertilization veliger 17h 24h
  15. 15. Effects of OA on Development =380 ppm =840 ppm Proportion Eggs in Cleavage *
  16. 16. Effects of OA on Development =380 ppm =840 ppm Proportion Larvae Hatched *
  17. 17. Effects of OA on Development   A smaller proportion of larvae in elevated pCO2 develop at the “normal” rate   Implications for physiological anomalies and calcification SEM Photo: Carla Stehr
  18. 18. Effects of OA on Development   General stress response genes   Cytochrome P450 oxidase (CytP450)   Peroxiredoxin 6 (Prx6)   Heat shock protein 70 (Hsp70) 1.0 0.8 Proportion Hatched 0.6 0.4 0.2 0.0 5 5 17 17 24 24 Time Post Fertilization (Hours)
  19. 19. Outline   Ocean acidification   Environmental stress and physiology   Temperature and the C. gigas-V. tubiashii relationship   OA effects on C. gigas   Current work: OA & V. tubiashii Hi OA T
  20. 20. OA & V. tubiashii   Study in progress in collaboration with NWFSC (P. McElhany et al.)   Planned 3 weeks: spawning through settlement   4 pCO2 treatments   280, 380, 750, 2000 ppm   24-hour V. tubiashii exposure   Data collection:   Mortality   Physiology (gene expression)   Morphology & calcification   Genotype-linked survival
  21. 21. = calcified OA & V. tubiashii = partially calcified   Calcification at 48 hours post-fertilization = uncalcified
  22. 22. Conclusions   Ocean acidification has significant effects on C. gigas larval development.   OA may inhibit and depress larval response to other environmental stressors, including disease.   Elevated temperatures, such as 25oC, have effects on the host-pathogen interaction.
  23. 23. Acknowledgements   Funding: Saltonstall-Kennedy Grant (NOAA), NOAA Aquaculture Program student support   University of Washington   Moose O’Donnell   Sam White   Taylor Shellfish Joth Davis Ed Jones Vicki Jones   NOAA Northwest Fisheries Science Center   Jason Miller   Mike Maher   Sarah Norberg

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