Journal of Comparative Physiology B

, Volume 182, Issue 1, pp 41–48

Immune response and mechanical stress susceptibility in diseased oysters, Crassostrea virginica

  • Steven B. Roberts
  • Inke Sunila
  • Gary H. Wikfors
Original Paper

Abstract

Eastern oysters, Crassostrea virginica, naturally infected with the parasite Perkinsus marinus were subjected to a mechanical stress by centrifugation, and immune parameters, pathological conditions, and gene expression of selected transcripts were compared to uninfected controls. Immune parameters were assessed by flow cytometry, pathology and parasites by histotechnology and fluid thioglycollate assays, and gene expression by quantitative RT-PCR. Irrespective of mechanical stress, an increased number of hemocytes were observed in P. marinus-infected oysters that corresponded to increased expression of genes that have been shown to be involved in inflammation and apoptosis, two processes associated with regulating immune cell populations. Mechanically stressed, diseased oysters showed histological gill abnormalities and aggregations of hemocytes in tissues not seen in stressed, uninfected oysters. Expression of a high-mobility group protein and hemocyte phagocytosis were significantly upregulated upon mechanical stress only in uninfected oysters. The results of this study demonstrate the role of inflammation in the oyster immune response including possible underlying molecular mechanisms. Furthermore, this study highlights the importance of considering mechanical stressors when characterizing oyster immune function.

Keywords

Oyster Stress Gene expression Immune Hemocyte 

References

  1. Ahmed H, Schott EJ, Gauthier JD, Vasta GR (2003) Superoxide dismutases from the oyster parasite Perkinsus marinus: purification, biochemical characterization, and development of a plate microassay for activity. Anal Biochem 318:132–141PubMedCrossRefGoogle Scholar
  2. Anderson RS (1996) Interactions of Perkinsus marinus with humoral factors and hemocytes of Crassostrea virginica. J Shellfish Res 15:127–134Google Scholar
  3. Andrews JD, Hewatt WG (1957) Oyster mortality studies in Virginia. II. The fungus disease caused by Dermocystidium marina in oysters in Chesapeake Bay. Ecol Monographs 27:1–26CrossRefGoogle Scholar
  4. Brousseau DJ (1996) Epizootiology of the parasite, Perkinsus marinus (Dermo) in intertidal oyster populations from Long Island Sound. J Shellfish Res 15:583–587Google Scholar
  5. Bushek D, Ford SE, Allen SK Jr (1994) Evaluation of methods using Ray’s fluid thioglycollate medium for the diagnosis of Perkinsus marinus infection in the eastern oyster, Crassostrea virginica. Annu Rev Fish Dis 4:201–217CrossRefGoogle Scholar
  6. Desmazes C, Galineau L, Gauthier F, Bromme D, Lalmanach G (2003) Kininogen-derived peptides for investigating the putative vasoactive properties of human cathepsins K and L. Eur J Biochem 270:171–178PubMedCrossRefGoogle Scholar
  7. Donald KM, Day AJ, Smerdon GR, Cross LJ, Hawkins AJS (2003) Quantification of gene transcription and enzyme activity for functionally important proteolytic enzymes during early development in the Pacific oyster Crassostrea gigas. Comp Biochem Physiol Part B Biochem Mol Biol 136:383–392CrossRefGoogle Scholar
  8. Dumitriu IE, Baruah P, Manfredi AA, Bianchi ME, Rovere-Querini P (2005) HMGB1: guiding immunity from within. Trends Immunol 26:381–387PubMedCrossRefGoogle Scholar
  9. Ford SE (1996) Range extension by the oyster parasite Perkinsus marinus into the northeastern United States: response to climate change? J Shellfish Res 15:45–56Google Scholar
  10. Ford SE, Haskin HH (1982) Histology and epizootiology of Haplosporidium nelsoni (MSX), an oyster pathogen, in Delaware Bay, 1957–1980. J Invert Pathol 40:118–141CrossRefGoogle Scholar
  11. Ford SE, Tripp MR (1996) Diseases and defense mechanisms. In: Kennedy VS, Newell RIE, Eble AF (eds) The eastern oyster, Crassostrea virginica. Maryland Sea Grant College, College Park, Maryland, pp 581–586Google Scholar
  12. Hégaret H, Wikfors GH, Soudant P (2003a) Flow-cytometric analysis of haemocytes from eastern oysters, Crassostrea virginica, subjected to a sudden elevation in temperature: I. Haemocyte types and morphology. J Exp Mar Biol Ecol 293:237–248CrossRefGoogle Scholar
  13. Hégaret H, Wikfors GH, Soudant P (2003b) Flow-cytometric analysis of hemocytes from eastern oysters, Crassostrea virginica, subjected to a sudden, sudden temperature elevation: II. Hemocyte functions: aggregation, viability, phagocytosis and respiratory burst. J Exp Mar Biol Ecol 293:249–265CrossRefGoogle Scholar
  14. Howard DW, Lewis EJ, Keller BJ, Smith CS (2004) Histological techniques for marine bivalve mollusks and crustaceans. NOAA Tech Memo NOS NCCOS 5:218Google Scholar
  15. Hughes FM, Foster B, Grewal S, Sokolova IM (2010) Apoptosis as a host defense mechanism in Crassostrea virginica and its modulation by Perkinsus marinus. Fish Shellfish Immunol. doi:10.1016/j.fsi.2010.03.003
  16. Karolus J, Sunila I, Spear S, Volk J (2000) Prevalence of Perkinsus marinus (Dermo) in Crassostrea virginica along the Connecticut shoreline. Aquaculture 183:215–221CrossRefGoogle Scholar
  17. Kleinschuster SJ, Parent J (1995) Sub-clinical infection of oysters (Crassostrea virginica) (Gmelin 1791) from Maine by species of the genus Perkinsus (Apicomplexa). J Shellfish Res 14:489–491Google Scholar
  18. Lacoste A, Malham SK, Cueff A, Poulet SA (2001) Stress-induced catecholamine changes in the hemolymph of the oyster Crassostrea gigas. Gen Comp Endocr 121:181–188CrossRefGoogle Scholar
  19. Lacoste A, Malham SK, Gélébart F, Cueff A, Poulet SA (2002) Stress-induced immune changes in the oyster Crassostrea gigas. Dev Comp Immunol 26(1):1–9PubMedCrossRefGoogle Scholar
  20. Liu L, Warner AH (2006) Further characterization of the cathepsin L-associated protein and its gene in two species of the brine shrimp, Artemia. Comp Biochem Physiol Part A Mol Integr Physiol 145:458–467CrossRefGoogle Scholar
  21. Mackin JG (1962) Oyster disease caused by Dermocystidium marinum and other microorganisms in Louisiana. Publ Inst Mar Sci Univ Tex 7:132–229Google Scholar
  22. Mackin JG, Owen HM, Collier A (1950) Preliminary note on the occurrence of a new protozoan parasite, Dermocystidium marinum n. sp. in Crassostrea virginica (Gmelin). Science 111:328–329PubMedCrossRefGoogle Scholar
  23. Oliver JL, Gaffney PM, Allen SK Jr, Faisal M, Kaattari SL (2000) Protease inhibitory activity in selectively bred families of eastern oysters, Crassostrea virginica. J Aquatic Ani Health 12:136–145CrossRefGoogle Scholar
  24. Perkins FO (1976) Dermocystidium marinum infection in oysters. Mar Fish Rev 38:19–21Google Scholar
  25. Sunila I, LaBanca J (2003) Apoptosis in the pathogenesis of infectious diseases of the eastern oyster Crassostrea virginica. Dis Aquatic Org 56:163–170CrossRefGoogle Scholar
  26. Vasta GR, Gauthier JD, Marsh AG (1995) Molecular and biochemical adaptations of the protozoan pathogen Perkinsus marinus for its host, Crassostrea virginica: current research and future perspectives (or “the best defense is a good offense”). J Mar Biotechnol 3:35–41Google Scholar
  27. Venier P, De Pittà C, Pallavicini A, Marsano F, Varotto L, Romualdi C, Dondero F, Viarengo A, Lanfranchi G (2006) Development of mussel mRNA profiling: can gene expression trends reveal coastal water pollution? Mutat Res 602(1–2):121–134Google Scholar
  28. Zhao S, Fernald RD (2005) Comprehensive algorithm for quantitative real-time polymerase chain reaction. J Comput Biol 12(8):1045–1062CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Steven B. Roberts
    • 1
  • Inke Sunila
    • 2
  • Gary H. Wikfors
    • 3
  1. 1.University of Washington, School of Aquatic and Fishery SciencesSeattleUSA
  2. 2.State of Connecticut, Department of AgricultureBureau of AquacultureMilfordUSA
  3. 3.Northeast Fisheries Science CenterNMFS, NOAAMilfordUSA

Personalised recommendations