Characterisation of the antiviral response associated with “mass mortalities” of the Pacific oysters, Crassostrea gigas Timothy Green, Caroline Montagnani,

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Presentation transcript:

Characterisation of the antiviral response associated with “mass mortalities” of the Pacific oysters, Crassostrea gigas Timothy Green, Caroline Montagnani, Nick Robinson, Kirsten Benkendorff & Peter Speck

Survival - why do oysters die? Pernet et al., 2013

IFN-like pathway in Oysters? Green & Montagnani 2013

Oysters have a complex antiviral immune repertoire!!! Green et al.,

Within-Generation Immune Priming Poly I:C: synthetic dsRNA, which mimics a virus.

Results: Trial I

Results: Trial II

Conclusion dsRNA (poly I:C) induces C. gigas into an antiviral state. Oyster possess a specific and inducible innate antiviral response.

Control Measures for POMS Farm management/husbandry o Gagnaire et al., 2006; Peeler et al., 2012; Pernet et al., 2012; Paul-Pont et al., o Successful management requires knowledge: Virus reservoir, routes of transmission, environmental risk factors, etc. o Survival cannot be at the detriment of other commercial traits!

Control Measures for POMS Farm management/husbandry o Gagnaire et al., 2006; Peeler et al., 2012; Pernet et al., 2012; Paul-Pont et al., o Successful management requires knowledge: Virus reservoir, routes of transmission, environmental risk factors, etc. o Survival cannot be at the detriment of other commercial traits! Develop POMS resistant oysters o Natural selection (> 10 years). o Translocation of broodstock between jurisdictions. o Indirect selection?

Control Measures for POMS Farm management/husbandry o Gagnaire et al., 2006; Peeler et al., 2012; Pernet et al., 2012; Paul-Pont et al., o Successful management requires knowledge: Virus reservoir, routes of transmission, environmental risk factors, etc. o Survival cannot be at the detriment of other commercial traits! Develop POMS resistant oysters o Natural selection (> 10 years). o Translocation of broodstock between jurisdictions. o Indirect selection? Trans-generational immune priming o ?

Evidence of trans-generational immune priming in invertebrates

Evidence of trans-generational immune priming in Pacific oysters to OsHV-1

Future Directions Questions: o Does transgenerational priming exist in Crassostrea gigas? o Can you prime with a virus associated molecular pattern (i.e. poly I:C) o If so, what is the molecular mechanism that provides increased protection to oyster progeny?

Inducible antiviral response in oysters? Results: Gene expression profiles measured didn’t explain the reduced viral load in the resistant family line. Question: Do oysters have an inducible innate immune response against virus infection?

Plaque-reduction assay as a method for screening antiviral activity Vero cells + HSV-1 Vero cells Vero cells + HSV-1 + oyster 1 Vero cells + HSV-1 + oyster 2 Vero cells Vero cells + HSV-1 + oyster 1 Vero cells + HSV-1 + Oyster 2

Antiviral Activity of Australian Pacific oyster hemolymph ASI Family Lines h 2 = 0.21

Identification of oyster antiviral compound(s)

Why do oysters die? Laboratory experiment 1)Treatment (poly I:C verse seawater) 2)Temperature (12 verse 22 o C) 3)Development (Adults verse Juvenile) Hypothesis:  Virus relies on faster cell replication rate of juvenile cells.  Therefore, temperature and developmental stage has no affect on immune response.  Alternative hypothesis: oysters at 22 o C are immuno-suppressed or the immune system of juveniles is immature.

Summary/Future Direction 1Developing a proxy for indirect selection. 2Why oysters die. 3Continue investigating trans-generational priming as a control mechanism.

POMS challenge trials in SA POMS challenge model would be advantageous to our research. We have asked SAOGA & PIRSA to conduct a POMS challenge trials at the physical containment (PC2) facility at Roseworthy. We will work with SAOGA & PIRSA to make sure this research is conducted safely. Our research will be overseen by the Flinder’s Institutional Biosafety Committee. We have experience conducting POMS challenge trials. We currently use HSV-1 as a surrogate model. Experience with human viruses. Project will build POMS research capacity in SA!!! It will be done safely!!!

Acknowledgements Seafood CRC (Graham Mair) ASI NSW Fisheries (Wayne) RIME (Julien, Delphine, Agnes) IFREMER (La Treblade, Palavas, Sete) SAOGA (Trudy) PIRSA & SARDI (Shane, Marty & Mehdi) Gary Zippel