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In vitro culture and developmental cycle of the parasitic dinoflagellate Hematodinium sp. from the blue crab Callinectes sapidus

Published online by Cambridge University Press:  09 September 2011

CAIWEN LI
Affiliation:
Virginia Institute of Marine Science, School of Marine Science, The College of William and Mary, P.O. Box 1346, Gloucester Point, Virginia 23062, USA
TERRENCE L. MILLER
Affiliation:
Virginia Institute of Marine Science, School of Marine Science, The College of William and Mary, P.O. Box 1346, Gloucester Point, Virginia 23062, USA
HAMISH J. SMALL
Affiliation:
Virginia Institute of Marine Science, School of Marine Science, The College of William and Mary, P.O. Box 1346, Gloucester Point, Virginia 23062, USA
JEFFREY D. SHIELDS*
Affiliation:
Virginia Institute of Marine Science, School of Marine Science, The College of William and Mary, P.O. Box 1346, Gloucester Point, Virginia 23062, USA
*
*Corresponding author: Tel: 001 804 684 7128. Fax: 001-804 684 7186. E-mail: [email protected]

Summary

Hematodinium is a genus of parasitic dinoflagellates whose species have caused significant mortalities in marine crustacean fisheries worldwide. A species of Hematodinium infects the blue crab, Callinectes sapidus on the eastern seaboard of the USA. The mode of transmission of the parasite in blue crabs is unknown. We established several continuous in vitro cultures of Hematodinium sp. isolated from the haemolymph of infected blue crabs. One isolate has been continuously maintained in our laboratory through serial subcultivation for over 12 months, and is capable of infecting new hosts when inoculated into healthy crabs. Cells of the parasite undergo characteristic developmental changes in vitro consistent with the identifiable stages of Hematodinium sp.: filamentous trophonts, amoeboid trophonts, arachnoid trophonts and sporonts, sporoblasts, prespores and dinospores (macrospores and microspores). Additionally, we describe an unusual shunt in the life cycle wherein presumptive schizonts derived from arachnoid sporonts developed into filamentous and arachnoid trophonts that can then initiate arachnoid sporonts in new cultures. This may explain the rapid proliferation of the parasite in blue crab hosts. We also found that temperature and light intensity affected the growth and development of the parasite in vitro.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2011

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References

REFERENCES

Appleton, P. L. and Vickerman, K. (1998). In vitro cultivation and developmental cycle in culture of a parasitic dinoflagellate (Hematodinium sp.) associated with mortality of the Norway lobster (Nephrops norvegicus) in British waters. Parasitology 116, 115130.CrossRefGoogle Scholar
Chatton, É. and Poisson, R. (1931). Sur l'existence, dans le sang des Crabes, de Péridiniens parasites: Hematodinium perezi n.g., n.sp. (Syndinidae). Comptes Rendus des Séances de la Société de Biologie Paris 105, 553557.Google Scholar
Eaton, W. D., Love, D. C., Botelho, C., Meyers, T. R., Imamura, K. and Koeneman, T. (1991). Preliminary results on the seasonality and life cycle of the parasitic dinoflagellate causing Bitter Crab Disease in Alaskan Tanner crabs (Chionoecetes bairdi). Journal of Invertebrate Pathology 57, 426434.CrossRefGoogle ScholarPubMed
Field, R. H. and Appleton, P. L. (1995). A Hematodinium-like dinoflagellate infection of the Norway lobster Nephrops norvegicus: observations on pathology and progression of infection. Diseases of Aquatic Organisms 22, 115128.CrossRefGoogle Scholar
Field, R. H. and Appleton, P. L. (1996). An indirect fluorescent antibody technique for the diagnosis of Hematodinium sp. infection of the Norway lobster, Nephrops norvegicus. Diseases of Aquatic Organisms 24, 199204.CrossRefGoogle Scholar
Frischer, M. E., Lee, R. F., Sheppard, M. A., Mauer, A., Rambow, F., Neumann, M., Brofft, J. E., Wizenmann, T. and Danforth, J. M. (2006). Evidence for a free-living life stage of the blue crab parasitic dinoflagellate, Hematodinium sp. Harmful Algae 5, 548557.CrossRefGoogle Scholar
Hudson, D. A. and Shields, J. D. (1994). Hematodinium australis n. sp., a parasitic dinoflagellate of the sand crab Portunus pelagicus from Moreton Bay, Australia. Diseases of Aquatic Organisms 19, 109119.CrossRefGoogle Scholar
Jensen, P. C., Califf, K., Lowe, V., Hauser, L. and Morado, J. F. (2010). Molecular detection of Hematodinium sp. in Northeast Pacific Chionoecetes spp. and evidence of two species in the Northern Hemisphere. Diseases of Aquatic Organisms 89, 155166.CrossRefGoogle ScholarPubMed
Li, C., Shields, J. D., Miller, T. L., Small, H. J., Pagenkopp, K. M. and Reece, K. S. (2010). Detection and quantification of the free-living stage of the parasitic dinoflagellate Hematodinium sp. in laboratory and environmental samples. Harmful Algae 9, 515521.CrossRefGoogle Scholar
Li, C., Wheeler, K. N. and Shields, J. D. (2011). Lack of transmission of Hematodinium sp. in the blue crab Callinectes sapidus through cannibalism. Diseases of Aquatic Organisms (in the Press).CrossRefGoogle ScholarPubMed
Messick, G. A. (1994). Hematodinium perezi infections in adult and juvenile blue crabs Callinectes sapidus from coastal bays of Maryland and Virginia, USA. Diseases of Aquatic Organisms 19, 7782.CrossRefGoogle Scholar
Messick, G. A., Jordan, S. J. and Van Heukelem, W. F. (1999). Salinity and temperature effects on Hematodinium sp. in the blue crab Callinectes sapidus. Journal of Shellfish Research 18, 657662.Google Scholar
Messick, G. A. and Shields, J. D. (2000). Epizootiology of the parasitic dinoflagellate Hematodinium sp. in the American blue crab Callinectes sapidus. Diseases of Aquatic Organisms 43, 139152.CrossRefGoogle ScholarPubMed
Meyers, T. R., Botelho, C., Koeneman, T. M., Short, S. and Imamura, K. (1990). Distribution of bitter crab dinoflagellate syndrome in southeast Alaskan tanner crabs, Chionoecetes bairdi. Diseases of Aquatic Organisms 9, 3743.CrossRefGoogle Scholar
Meyers, T. R., Koeneman, T. M., Botelho, C. and Short, S. (1987). Bitter crab disease: a fatal dinoflagellate infection and marketing problem from Alaskan tanner crabs, Chionoecetes bairdi. Diseases of Aquatic Organisms 3, 195216.CrossRefGoogle Scholar
Newman, M. W. and Johnson, C. A. (1975). A disease of blue crabs (Callinectes sapidus) caused by a parasitic dinoflagellate, Hematodinium sp. Journal of Parasitology 63, 554557.CrossRefGoogle Scholar
Shields, J. D. (1994). The parasitic dinoflagellates of marine crustaceans. Annual Review Fish Diseases 4, 241271.CrossRefGoogle Scholar
Shields, J. D. (2003). Research priorities for diseases of the blue crab Callinectes sapidus. Bulletin of Marine Science 72, 505517.Google Scholar
Shields, J. D. and Overstreet, R. M. (2007). Diseases, parasites, and other symbionts. In The Blue Crab Callinectes sapidus (ed. Kennedy, V. S. and Cronin, L. E.), pp. 299394. Maryland Sea Grant College, College Park, MD, USA.Google Scholar
Shields, J. D. and Squyars, C. M. (2000). Mortality and hematology of blue crabs, Callinectes sapidus, experimentally infected with the parasitic dinoflagellate Hematodinium perezi. Fish Bulletin 98, 139152.Google Scholar
Shields, J. D., Taylor, D. M., Sutton, S. G., O'Keefe, P. O., Collins, P. W., Ings, D. W. and Pardy, A. L. (2005). Epizootiology of bitter crab disease (Hematodinium sp.) in snow crabs, Chionoecetes opilio, from Newfoundland, Canada. Diseases of Aquatic Organisms 64, 253264.CrossRefGoogle Scholar
Shields, J. D., Taylor, D. M., O'Keefe, P. G., Colbourne, E. and Hynick, E. (2007). Epidemiological determinants in outbreaks of bitter crab disease (Hematodinium sp.) in snow crabs, Chionoecetes opilio from Newfoundland, Canada. Diseases of Aquatic Organisms 77, 6172.CrossRefGoogle ScholarPubMed
Small, H. J. (2004). Infections of the Norway lobster Nephrops norvegicus (L) by dinoflagellate and ciliate parasites. Ph.D. thesis, University of Glasgow, Glasgow, UK.Google Scholar
Small, H. J., Shields, J. D., Hudson, K. L. and Reece, K. S. (2007 a). Molecular detection of Hematodinium sp. infecting the blue crab Callinectes sapidus. Journal of Shellfish Research 26, 131139.CrossRefGoogle Scholar
Small, H. J., Shields, J. D., Neil, D. M., Taylor, A. C. and Coombs, G. H. (2007 b). Differences in enzyme activities between two species of Hematodinium, parasitic dinoflagellates of crustaceans. Journal of Invertebrate Pathology 94, 175183.CrossRefGoogle ScholarPubMed
Stentiford, G. D., Neil, D. M. and Atkinson, R. J. A. (2001). The relationship of Hematodinium infection prevalence in a Scottish Nephrops norvegicus population to seasonality, moulting and sex. ICES Journal of Marine Sciences 58, 814823.CrossRefGoogle Scholar
Stentiford, G. D., Neil, D. M. and Coombs, G. H. (1999). Changes in the plasma free amino acid profile of the Norway lobster Nephrops norvegicus at different stages of infection by a parasitic dinoflagellate (genus Hematodinium). Disease of Aquatic Organisms 38, 151157.CrossRefGoogle Scholar
Stentiford, G. D., Neil, D. M. and Coombs, G. H. (2000). Alterations in the biochemistry and ultrastructure of the deep abdominal flexor muscle of the Norway lobster, Nephrops norvegicus L. during infection by a parasitic dinoflagellate of the genus Hematodinium. Disease of Aquatic Organisms 42, 133141.CrossRefGoogle ScholarPubMed
Stentiford, G. D. and Shields, J. D. (2005). A review of the parasitic dinoflagellates Hematodinium species and Hematodinium-like infections in marine crustaceans. Diseases of Aquatic Organisms 66, 4770.CrossRefGoogle ScholarPubMed
Taylor, A. C., Field, R. H. and Parslow-Williams, P. J. (1996). The effects of Hematodinium sp. – infection on aspects of the respiratory physiology of the Norway lobster, Nephrops norvegicus (L.). Journal of Experimental Marine Biology and Ecology 207, 217228.CrossRefGoogle Scholar
Wheeler, K., Shields, J. D. and Taylor, D. M. (2007). Pathology of Hematodinium sp. infections in snow crabs (Chionoecetes opilio) from Newfoundland, Canada. Journal of Invertebrate Pathology 95, 93100.CrossRefGoogle ScholarPubMed