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Cytotoxicity of quahog parasite unknown (QPX) toward hard clam (Mercenaria mercenaria) haemocytes and interactions between different pathogen isolates and host strains

Published online by Cambridge University Press:  23 July 2009

MICKAEL PERRIGAULT
Affiliation:
School of Marine and Atmospheric Sciences, Stony Brook University, Stony Brook. NY 11794-5000, USA
BASSEM ALLAM*
Affiliation:
School of Marine and Atmospheric Sciences, Stony Brook University, Stony Brook. NY 11794-5000, USA
*
*Corresponding author: Tel: +1 631 632 8745. Fax: +1 631 632 3072. E-mail: [email protected]

Summary

The ability of pathogens to neutralize host defence mechanisms represents a fundamental requisite in the successful establishment of an infection. Host-pathogen interactions between quahog parasite unknown (QPX) and its hard clam host are poorly understood. Our prior in vivo investigations have shown that different QPX isolates display varying levels of pathogenicity toward clams. Similarly, field investigations and laboratory transmission studies revealed some variations in the susceptibility of different hard clam stocks to QPX infection. An in vitro approach was developed in this study to evaluate the toxicity of QPX cells and extracellular products toward haemocytes using a neutral red uptake assay. Results demonstrated that QPX produces virulence factors that are cytotoxic to M. mercenaria haemocytes. This cytotoxicity appears to be induced by clam factors, suggesting that it may play an important role in supporting QPX infection and proliferation within the host. Moreover, application of this technique to different QPX isolates and clam broodstocks indicates variations of QPX cytotoxicity in agreement with previous in vivo experiments, strengthening the existence of different QPX strains.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2009

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References

REFERENCES

Allam, B., Paillard, C. and Ford, S. E. (2002). Pathogenicity of Vibrio tapetis, the etiological agent of brown ring disease in clams. Diseases of Aquatic Organisms 48, 221231.CrossRefGoogle ScholarPubMed
Anderson, R. S., Kraus, B. S., McGladdery, S. and Smolowitz, R. (2003). QPX, a pathogen of quahogs (hard clams), employs mucoid secretions to resist host antimicrobial agents. Journal of Shellfish Research 22, 205208.Google Scholar
Anderson, R. S., Luskey, T. M. and Strauss, M. A. (2006). In vitro protease production by QPX (Abstract). Journal of Shellfish Research 25, 708.Google Scholar
Borenfreund, E. and Puerner, J. A. (1985 a). A simple quantitative procedure using monolayer cultures for cytoxicity assays (HTD/NR-90). Journal of Tissue Culture Methods 9, 79.CrossRefGoogle Scholar
Borenfreund, E. and Puerner, J. A. (1985 b). Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicology Letters 24, 119124.CrossRefGoogle ScholarPubMed
Bower, S. M. (1987). Pathogenicity and host specificity of Labyrinthuloides haliotidis (Protozoa, Labyrinthomorpha), a parasite of juvenile abalone. Canadian Journal of Zoology 65, 20082012.CrossRefGoogle Scholar
Buggé, D. M. and Allam, B. (2005). A fluorometric technique for the in vitro measurement of growth and viability in Quahog parasite unknown (QPX). Journal of Shellfish Research 24, 10131018.Google Scholar
Bushek, D. and Allen, S. K. (1996 a). Host-parasite interactions among broadly distributed populations of the eastern oyster Crassostrea virginica and the protozoan Perkinsus marinus. Marine Ecology-Progress Series 139, 127141.CrossRefGoogle Scholar
Bushek, D. and Allen, S. K. (1996 b). Races of Perkinsus marinus. Journal of Shellfish Research 15, 103107.Google Scholar
Cajaraville, M. P., Olabarrieta, I. and Marigomez, I. (1996). In vitro activities in mussel hemocytes as biomarkers of environmental quality: A case study in the Abra Estuary (Biscay Bay). Ecotoxicology and Environmental Safety 35, 253260.CrossRefGoogle ScholarPubMed
Calvo, L. M. R., Ford, S. E., Kraeuter, J. N., Leavitt, D. F., Smolowitz, R. and Burreson, E. M. (2007). Influence of host genetic origin and geographic location on QPX disease in Northern quahogs (=hard clams), Mercenaria mercenaria. Journal of Shellfish Research 26, 109119.CrossRefGoogle Scholar
Calvo, L. M. R., Walker, J. G. and Burreson, E. M. (1998). Prevalence and distribution of QPX, Quahog Parasite Unknown, in hard clams Mercenaria mercenaria in Virginia, USA. Diseases of Aquatic Organisms 33, 209219.CrossRefGoogle Scholar
Choquet, G., Soudant, P., Lambert, C., Nicolas, J. L. and Paillard, C. (2003). Reduction of adhesion properties of Ruditapes philippinarum hemocytes exposed to Vibrio tapetis. Diseases of Aquatic Organisms 57, 109116.CrossRefGoogle ScholarPubMed
Chu, F. L. E., Volety, A. K., Hale, R. C. and Huang, Y. Q. (2002). Cellular responses and disease expression in oysters (Crassostrea virginica) exposed to suspended field – contaminated sediments. Marine Environmental Research 53, 1735.CrossRefGoogle ScholarPubMed
Coles, J. A., Farley, S. R. and Pipe, R. K. (1995). Alteration of the immune response of the common marine mussel Mytilus edulis resulting from exposure to cadmium. Diseases of Aquatic Organisms 22, 5965.CrossRefGoogle Scholar
Dahl, S., Perrigault, M. and Allam, B. (2008). Laboratory transmission studies of QPX disease in the hard clam: Interactions between different host strains and pathogen isolates. Aquaculture 280, 6470.CrossRefGoogle Scholar
Dove, A. D. M., Bowser, P. R. and Cerrato, R. M. (2004). Histological analysis of an outbreak of QPX disease in wild hard clams Mercenaria mercenaria in New York. Journal of Aquatic Animal Health 16, 246250.CrossRefGoogle Scholar
Earnhart, C. G., Vogelbein, M. A., Brown, G. D., Reece, K. S. and Kaattari, S. L. (2004). Supplementation of Perkinsus marinus cultures with host plasma or tissue homogenate enhances their infectivity. Applied and Environmental Microbiology 70, 421431.CrossRefGoogle ScholarPubMed
Ford, S. E. (2001). Pests, parasites, diseases and defense mechanisms of the hard clam, Mercenaria mercenaria. In Biology of the Hard Clam, (ed. Kraeuter, J. N. and Castagna, M.), pp. 591628. Elsevier Science, Amsterdam, The Netherlands.CrossRefGoogle Scholar
Ford, S. E., Kraeuter, J. N., Barber, R. D. and Mathis, G. (2002). Aquaculture-associated factors in QPX disease of hard clams: density and seed source. Aquaculture 208, 2338.CrossRefGoogle Scholar
Garreis, K. A., La Peyre, J. F. and Faisal, M. (1996). The effects of Perkinsus marinus extracellular products and purified proteases on oyster defence parameters in vitro. Fish and Shellfish Immunology 6, 581597.CrossRefGoogle Scholar
Grundy, M. M., Moore, M. N., Howell, S. M. and Ratcliffe, N. A. (1996). Phagocytic reduction and effects on lysosomal membranes by polycyclic aromatic hydrocarbons, in haemocytes of Mytilus edulis. Aquatic Toxicology 34, 273290.CrossRefGoogle Scholar
Jain, R., Raghukumar, S., Tharanathan, R. and Bhosle, N. B. (2005). Extracellular polysaccharide production by thraustochytrid protists. Marine Biotechnology 7, 184192.CrossRefGoogle ScholarPubMed
Kleinschuster, S. J., Smolowitz, R. and Parent, J. (1998). In vitro life cycle and propagation of quahog parasite unknown. Journal of Shellfish Research 17, 7578.Google Scholar
Lane, E. and Birkbeck, T. H. (2000). Species specificity of some bacterial pathogens of bivalve molluscs is correlated with their interaction with bivalve haemocytes. Journal of Fish Diseases 23, 275279.CrossRefGoogle Scholar
Le Sueur, L. D., Collares-Buzato, C. B., Kalapothakis, E. and Da Cruz-Hofling, M. A. (2005). In vitro effect of the Phoneutria nigriventer spider venom on cell viability, paracellular barrier function and transcellular transport in cultured cell lines. Toxicon 46, 130141.CrossRefGoogle Scholar
Maas, P. A. Y., Kleinschuster, S. J., Dykstra, M. J., Smolowitz, R. and Parent, J. (1999). Molecular characterization of QPX (Quahog Parasite Unknown), a pathogen of Mercenaria mercenaria. Journal of Shellfish Research 18, 561567.Google Scholar
MacIntyre, E. A., Earnhart, C. G. and Kaattari, S. L. (2003). Host oyster tissue extracts modulate in vitro protease expression and cellular differentiation in the protozoan parasite, Perkinsus marinus. Parasitology 126, 293302.CrossRefGoogle ScholarPubMed
Matozzo, V., Ballarin, L. and Marin, M. G. (2002). In vitro effects of tributyltin on functional responses of haemocytes in the clam Tapes philippinarum. Applied Organometallic Chemistry 16, 169174.CrossRefGoogle Scholar
Muehlstein, L. K., Porter, D. and Short, F. T. (1988). Labyrinthula sp., a marine slime-mold producing the symptoms of wasting disease in eelgrass, Zostera marina. Marine Biology 99, 465472.CrossRefGoogle Scholar
Perrigault, M., Buggé, D. M., Hao, C. C. and Allam, B. (2009). Modulatory effects of hard clam (Mercenaria mercenaria) tissue extracts on the in vitro growth of its pathogen QPX. Journal of Invertebrate Pathology 100, 18.CrossRefGoogle ScholarPubMed
Pichardo, S., Jos, A., Zurita, J., Salguero, M., Camean, A. and Repetto, G. (2006). Toxic effects produced by microcystins from a natural cyanobacterial bloom and a Microcystis aeruginosa isolated strain on the fish cell lines RTG-2 and PLHC-1. Archives of Environmental Contamination and Toxicology 51, 8696.CrossRefGoogle Scholar
Pipe, R. K., Coles, J. A., Carissan, F. M. M. and Ramanathan, K. (1999). Copper induced immunomodulation in the marine mussel, Mytilus edulis. Aquatic Toxicology 46, 4354.CrossRefGoogle Scholar
Qian, H., Liu, Q. Q., Allam, B. and Collier, J. L. (2007). Molecular genetic variation within and among isolates of QPX (Thraustochytridae), a parasite of the hard clam Mercenaria mercenaria. Diseases of Aquatic Organisms 77, 159168.CrossRefGoogle ScholarPubMed
Ragan, M. A., MacCallum, G. S., Murphy, C. A., Cannone, J. J., Gutell, R. R. and McGladdery, S. E. (2000). Protistan parasite QPX of hard shell clam Mercenaria mercenaria is a member of Labyrinthulomycota. Diseases of Aquatic Organisms 42, 185190.CrossRefGoogle ScholarPubMed
Raghukumar, S. (2002). Ecology of the marine protists, the Labyrinthulomycetes (Thraustochytrids and Labyrinthulids). European Journal of Protistology 38, 127145.CrossRefGoogle Scholar
Reece, K. S., Bushek, D., Hudson, K. L. and Graves, J. E. (2001). Geographic distribution of Perkinsus marinus genetic strains along the Atlantic and Gulf coasts of the USA. Marine Biology 139, 10471055.Google Scholar
Smolowitz, R., Leavitt, D. and Perkins, F. (1998). Observations of a protistan disease similar to QPX in Mercenaria mercenaria (hard clams) from the coast of Massachusetts. Journal of Invertebrate Pathology 71, 9–25.CrossRefGoogle ScholarPubMed
Stokes, N. A., Calvo, L. M. R., Reece, K. S. and Burreson, E. M. (2002). Molecular diagnostics, field validation, and phylogenetic analysis of Quahog Parasite Unknown (QPX), a pathogen of the hard clam Mercenaria mercenaria. Diseases of Aquatic Organisms 52, 233247.CrossRefGoogle ScholarPubMed
Whyte, S. K., Cawthorn, R. J. and McGladdery, S. E. (1994). QPX (Quahaug Parasite X), a pathogen of northern quahaug Mercenaria mercenaria from the gulf of St Lawrence, Canada. Diseases of Aquatic Organisms 19, 129136.CrossRefGoogle Scholar
Zaroogian, G., Anderson, S. and Voyer, R. A. (1992). Individual and combined cytotoxic effects of cadmium, copper, and nickel on brown cells of Mercenaria mercenaria. Ecotoxicology and Environmental Safety 24, 328337.CrossRefGoogle ScholarPubMed