1. CHARACTERIZING DISEASE RESISTANCE IN NATIVE OYSTERSThat Have Experienced Disease Pressure Roxanna Smolowitz, Steven Roberts, Jackie DeFaveri Christina Romano, Rick Karney

2. Oyster Disease

Oyster disease is a significant issue

• Dermo ( Perkinsus marinus )

• MSX ( Haplosporidium nelsoni )

• JOD ( Roseabacter sp .)

Disease tolerance / resistance strains of oysters have been developed ( i.e. Rutgers )

In times of massive disease outbreaks, a limited number of oysters commonly survive

3. Research Objectives

Demonstrate seed originating from local wild oysters, that have experienced heavy disease (Dermo) pressure, could significantly contribute to the development of disease resistance

Genetically characterize regional oysters that are putatively resistant to Dermo

• develop genetic markers

• better understand mechanisms involved in immunity

4. Oysters that had experienced disease pressure Oysters that had not experienced disease pressure (nave) Spawn Spawn Experimental Design 5. Oysters (~4000 from each group) were grown in a Dermo prone site- Edgartown Great Pond July 2005 - Present 6. overwintered on bottom Temperature Data 7. Mortality Nave oysters Survivors ofDiseasePressure 8. Survivors of Dermo pressure [Edgartown GP] Naive [Tisbury] 9. DERMO Perkinsus marinus Survivors of Dermo pressure [Edgartown GP] Naive [Tisbury] 10. Research Objectives

Demonstrate seed originating from local wild oysters, that have experienced heavy disease (Dermo) pressure, could significantly contribute to the development of disease resistance

Genetically characterize regional oysters that are putatively resistant to Dermo

• develop genetic markers

• better understand mechanisms involved in immunity

11. Photo: Gary H. Wikfors: Northeast Fisheries Science Center,NOAA Fisheries Milford Laboratory, Milford, CT Immune Response in Oysters 12. What are the biochemical orgene productsproduced by hemocytes? 13. Approach: Genetic Characterization

Extract RNA from hemocyte rich tissue- gills (n=10)

Perform qRT-PCR for genes suspected to play a role in immune response

Normalize expression data to 18s RNA levels

14. CIAPIN Expression cytokine induced apoptosis inhibitor 1 Survivors of Dermo pressure [Edgartown GP] Naive [Tisbury] Apoptosis (programmed cell death) regulationis likely involved in disease resistance 15. Survivors of Dermo pressure [Edgartown GP] Naive [Tisbury] Cathepsin L Expression Cathepsins are a family of proteasesProteases and protease inhibitors have been shown to play critical roles in immunity and host-parasite interactions.One possibility is Cathepsin L is involved inP. marinusdefense activity. 16. For more information. 17. Conclusions

Seed originating from local wild oysters, that have experienced heavy disease (Dermo) pressure, could significantly contribute to the development of disease resistance in cultured oysters.

Our initial gene expression data indicates that there are differences in immune response, however more work is required to elucidate the complex processes involved.