Continuous culture of Perkinsus mediterraneus, a parasite of the European flat oyster Ostrea edulis, and characterization of its morphology, propagation, and extracellular proteins in vitro

Journal of Eukaryotic Microbiology

Volumn 55, Issue 1, 2008, Pages 34-43

  Casas, Sandra M ^a   Reece, Kimberly S ^b   Li, Yanli ^a   Moss, Jessica A ^b   Villalba, Antonio ^c   La Peyre, Jerome F ^a,d  

^a LOUISIANA STATE UNIVERSITY AGRICULTURAL CENTER   (United States)

^b VIRGINIA INSTITUTE OF MARINE SCIENCE   (United States)

^c CENTRO DE INVESTIGACIÓNS MARIÑAS   (Spain)

^d LOUISIANA STATE UNIVERSITY   (United States)

Author keywords

Actin; Bivalve mollusk; Extracellular enzymes; In vitro proliferation; ITS; LSU; Protistan parasite; rRNA; Serine proteases

Indexed keywords

ACTIN; ENZYME; PEPTIDE HYDROLASE; PROTOZOAL DNA; PROTOZOAL PROTEIN; RIBOSOME DNA; RIBOSOME RNA;

BIVALVE; ENZYME ACTIVITY; GENETIC ANALYSIS; MORPHOLOGY; PARASITE; PROTEIN; PROTIST; VIABILITY;

ANIMAL; ANIMAL PARASITOSIS; ARTICLE; CHEMISTRY; CLASSIFICATION; CULTURE TECHNIQUE; CYTOLOGY; DNA SEQUENCE; EUROPE; GENETICS; METABOLISM; OYSTER; PARASITOLOGY; PHYLOGENY; PHYSIOLOGY; POLYACRYLAMIDE GEL ELECTROPHORESIS; POLYMERASE CHAIN REACTION; PROTOZOON;

ACTINS; ANIMALS; CELL CULTURE TECHNIQUES; DNA, PROTOZOAN; DNA, RIBOSOMAL; ELECTROPHORESIS, POLYACRYLAMIDE GEL; ENZYMES; EUROPE; OSTREIDAE; PEPTIDE HYDROLASES; PHYLOGENY; POLYMERASE CHAIN REACTION; PROTOZOA; PROTOZOAN INFECTIONS, ANIMAL; PROTOZOAN PROTEINS; RNA, RIBOSOMAL; SEQUENCE ANALYSIS, DNA;

BIVALVIA; MOLLUSCA; OSTREA EDULIS; OSTREIDAE; PERKINSUS; PERKINSUS MEDITERRANEUS; PROTISTA;

EID: 38649105775     PISSN: 10665234     EISSN: 15507408     Source Type: Journal    
DOI: 10.1111/j.1550-7408.2008.00301.x     Document Type: Article

References (64)

1. Abollo, E., Casas, S. M., Ceschia, G. Villalba, A. 2006. Differential diagnosis of Perkinsus species by polymerase chain reaction-restriction fragment length polymorphism assay. Mol. Cell Probes, 20 : 323 329.
2. Ahmed, H., Schott, E. J., Gauthier, J. D. Vasta, G. R. 2003. Superoxide dismutases from the oyster parasite Perkinsus marinus: purification, biochemical characterization, and development of a plate microassay for activity. Anal. Biochem., 318 : 132 141.
3. Altschul, S. F., Gish, W., Miller, W., Myers, E. W. Lipman, D. J. 1990. Basic local alignment search tool. J. Mol. Biol., 215 : 403 410.
4. Anderson, R. S. 1999. Perkinsus marinus secretory products modulate superoxide anion production by oyster (Crassostrea virginica) haemocytes. Fish Shellfish Immunol., 9 : 51 60.
5. Audemard, C., Reece, K. S. Burreson, E. M. 2004. Real-time PCR for the detection and quantification of the protistan parasite Perkinsus marinus in environmental waters. Appl. Environ. Microbiol., 70 : 6611 6618.
6. Azevedo, C. 1989. Fine structure of Perkinsus atlanticus n. sp. (Apicomplexa, Perkinsea) parasite of the clam Ruditapes decussatus from Portugal. J. Parasitol., 75 : 627 635.
7. Benyon, R. J. Salvesen, G. 1989. Commercially available protease inhibitors. In : Benyon, R. J. Bond, J. S. (ed.), Proteolytic Enzymes: A Practical Approach. IRL Press, Oxford. APP. III : 241 249.
8. Burreson, E. M. Ragone Calvo, L. M. 1996. Epizootiology of Perkinsus marinus disease of oysters in Chesapeake Bay, with emphasis on data since 1985. J. Shellfish Res., 15 : 17 34.
9. Burreson, E. M., Reece, K. S. Dungan, C. F. 2005. Molecular, morphological and experimental evidence support synonymy of Perkinsus chesapeaki and Perkinsus andrewsi. J. Eukaryot. Microbiol., 52 : 258 270.
10. Burreson, E. M., Ragone-Calvo, L., La Peyre, J. F., Counts, F. Paynter, K. 1994. Acute osmotic tolerance of cultured cells of the oyster pathogen Perkinsus marinus (Apicomplexa: Perkinsida). Comp. Biochem. Physiol. A, 109 : 575 582.
11. Calvo, G. W. Burreson, E. M. 1994. In vitro and in vivo effects of eight chemotherapeutants on the oyster parasite Perkinsus marinus (Mackin, Owen, and Collier). J. Shellfish Res., 13 : 101 107.
12. Canestri-Trotti, G., Baccarani, E. M., Paesanti, F. Turolla, E. 2000. Monitoring of infections by protozoa of the genera Nematopsis, Perkinsus and Porospora in the smooth venus clam Callista chione from the North-Western Adriatic Sea (Italy). Dis. Aquat. Org., 42 : 157 161.
13. Casas, S. M. 2002. Estudio de la perkinsosis en la almeja fina, Tapes decussatus (Linnaeus 1758), de Galicia. Dissertation. University of Santiago de Compostela, Spain. 272 p.
14. Casas, S. M., Audemard, C. La Peyre, J. 2005. Comparison of Perkinsus marinus, Perkinsus chesapeaki and Perkinsus olseni infections in Eastern oyster (Crassostrea virginica). J. Shellfish Res., 24 : 645.
15. Casas, S. M., La Peyre, J. F., Reece, K. S., Azevedo, C. Villalba, A. 2002. Continuous in vitro culture of the carpet shell clam Tapes decussatus protozoan parasite Perkinsus atlanticus. Dis. Aquat. Org., 52 : 217 231.
16. Casas, S. M., Grau, A., Reece, K. S., Apakupakul, K., Azevedo, C. Villalba, A. 2004. Perkinsus mediterraneus n. sp. a protistan parasite of the European flat oyster Ostrea edulis (L.) from the coast of Balearic Islands, Mediterranean Sea. Dis. Aquat. Org., 58 : 231 244.
17. Chu, F. L. E., Lund, E., Soudant, P. Harvey, E. 2002. De novo arachidonic acid synthesis in Perkinsus marinus, a protozoan parasite of the eastern oyster Crassostrea virginica. Mol. Biochem. Parasitol., 119 : 179 190.
18. Coss, C. A., Robledo, J. A. F. Vasta, G. R. 2001. Fine structure of clonally propagated in vitro life stages of a Perkinsus sp. isolated from the Baltic clam Macoma balthica. J. Eukaryot. Microbiol., 48 : 38 51.
19. Da Ros, L. Canzonier, W. J. 1985. Perkinsus, a protistan threat to bivalve culture in the Mediterranean basin. Bull. Eur. Assoc. Fish Pathol., 5 : 23 25.
20. Dungan, C. F. Reece, K. S. 2006. In vitro propagation of two Perkinsus spp. parasites from Japanese Manila clams Venerupis philippinarum and description of Perkinsus honshuensis n. sp. J. Eukaryot. Microbiol., 53 : 316 326.
21. Dungan, C. F., Reece, K. S., Hamilton, R. M., Stokes, N. A. Burreson, E. M. 2007a. Experimental cross-infections by Perkinsus marinus and P. chesapeaki in three sympatric species of Chesapeake Bay oysters and clams. Dis. Aquat. Org., 76 : 67 75.
22. Dungan, C. F., Reece, K. S., Moss, J. A., Hamilton, R. M. Diggles, B. K. 2007b. Perkinsus olseni in vitro isolates from the New Zealand Clam Austrovenus stutchburyi. J. Eukaryot. Microbiol., 54 : 263 270.
23. Elandalloussi, L. M., Leite, R. B., Rodrigues, P. M., Afonso, R., Nunes, P. A. Cancela, M. L. 2005a. Effect of antiprotozoal drugs on the proliferation of the bivalve parasite Perkinsus olseni. Aquaculture, 243 : 9 17.
24. Elandalloussi, L. M., Rodrigues, P. M., Afonso, R., Leite, R. B., Nunes, P. A. Cancela, M. L. 2005b. Shikimate and folate pathways in the protozoan parasite, Perkinsus olseni. Mol. Biochem. Parasitol., 142 : 106 109.
25. Faisal, M., Schafhauser, D. Y., Garreis, K. A., Elsayed, E. La Peyre, J. F. 1999. Isolation and characterization of Perkinsus marinus proteases using bacitracin-sepharose affinity chromatography. Comp. Biochem. Physiol. B, 123 : 417 426.
26. Ford, S. E. 1996. Range extension by the oyster parasite Perkinsus marinus into the Northeastern United States: response to climate change? J. Shellfish Res., 15 : 45 56.
27. Garreis, K. A., La Peyre, J. F. Faisal, M. 1996. The effects of Perkinsus marinus extracellular products and purified proteases on oyster defence parameters in vitro. Fish Shellfish Immunol., 6 : 581 597.
28. Goggin, C. L. 1992. Occurrence of parasites of the genus Perkinsus in France. Bull. Eur. Ass. Fish Pathol., 12 : 174 176.
29. Goggin, C. L. Lester, R. J. G. 1987. Occurrence of Perkinsus species (Protozoa, Apicomplexa) in bivalves from the Great Barrier Reef. Dis. Aquat. Org., 3 : 113 117.
30. La Peyre, J. F. Faisal, M. 1997. Extracellular proteins of the oyster pathogen Perkinsus marinus as virulence factors and potential targets for therapy. J. Shellfish Res., 16 : 269.
31. La Peyre, J. F., Faisal, M. Burreson, E. M. 1993. In vitro propagation of the protozoan Perkinsus marinus, a pathogen of the eastern oyster, Crassostrea virginica. J. Eukaryot. Microbiol., 40 : 304 310.
32. La Peyre, J. F., Yarnall, H. A. Faisal, M. 1996. Contribution of Perkinsus marinus extracellular products in the infection of eastern oysters (Crassostrea virginica). J. Invertebr. Pathol., 68 : 312 313.
33. La Peyre, J. F., Schafhauser, D. Y., Rizkalla, E. H. Faisal, M. 1995. Production of serine proteases by the oyster pathogen Perkinsus marinus (Apicomplexa) in vitro. J. Eukaryot. Microbiol., 42 : 544 551.
34. La Peyre, M., Casas, S. La Peyre, J. 2006. Salinity effects on viability metabolic activity and proliferation of three Perkinsus species. Dis. Aquat. Org., 71 : 59 74.
35. La Peyre, M. K., Nickens, A. M., Volety, A. K., Tolley, G. S. La Peyre, J. F. 2003. Environmental significance of freshets in reducing Perkinsus marinus infection in eastern oysters Crassostrea virginica: potential management applications. Mar. Ecol. Prog. Ser., 248 : 165 176.
36. Lenaers, G., Maroteaux, L., Michot, B. Herzog, M. 1989. Dinoflagellates in evolution. A molecular phylogenetic analysis of large subunit ribosomal RNA. J. Mol. Evol., 29 : 40 51.
37. Lester, R. J. G. Davis, G. H. G. 1981. A new Perkinsus species (Apicomplexa, Perkinsea) from the abalone Haliotis ruber. J. Invertebr. Pathol., 37 : 181 187.
38. Lund, E. D., Chu, F. L. E., Soudant, P. Harvey, E. 2007. Perkinsus marinus, a protozoan parasite of the eastern oyster, has a requirement for dietary sterols. Comp. Biochem. Phys. A, 146 : 141 147.
39. Mackin, J. G. 1951. Histopathology of infection of Crassostrea virginica (Gmelin) by Dermocystidium marinum Mackin, Owen and Collier. Bull. Mar. Sci. Gulf Carib., 1 : 72 87.
40. Moss, J. A., Burreson, E. M. Reece, K. S. 2006. Advanced Perkinsus marinus infections in Crassostrea ariakensis maintained under laboratory conditions. J. Shellfish Res., 25 : 65 72.
41. Nickens, A. D., La Peyre, J. F., Wagner, E. S. Tiersch, T. R. 2002. An improved procedure to count Perkinsus marinus in eastern oyster hemolymph. J. Shellfish Res., 21 : 725 732.
42. O'Donoghue, P. J., Phillips, P. H. Shepherd, S. A. 1991. Perkinsus (Protozoa: Apicomplexa) infections in abalone from South Australian waters. Trans. R. Soc. South Aust., 115 : 77 82.
43. O'Farrell, C. L., La Peyre, J. F., Paynter, K. T. Burreson, E. M. 2000. Osmotic tolerance and volume regulation in in vitro cultures of the oyster pathogen Perkinsus marinus. J. Shellfish Res., 19 : 139 145.
44. Park, K. I., Choi, K. S. Choi, J. W. 1999. Epizootiology of Perkinsus sp. found in the Manila clam Ruditapes philippinarum in Komsoe Bay, Korea. J. Korean Fish Soc., 32 : 303 309.
45. Park, K. I., Park, J. K., Lee, J. Choi, K. S. 2005. Use of molecular markers for species identification of Korean Perkinsus sp. isolated from Manila clams Ruditapes philippinarum. Dis. Aquat. Org., 66 : 255 263.
46. Perkins, F. O. 1996. The structure of Perkinsus marinus (Mackin, Owen and Collier 1950) Levine, 1978 with comments on taxonomy and phylogeny of Perkinsus spp. J. Shellfish Res., 15 : 67 87.
47. Ray, S. M. 1952. A culture technique for the diagnosis of infection with Dermocystidium marinum Mackin, Owen, and Collier in oysters. Science, 116 : 360 361.
48. Ray, S. M., Mackin, J. G. Boswell, J. L. 1953. Quantitative measurement of the effect on oysters of disease caused by Dermocystidium marinum. Bull. Mar. Sci. Gulf Caribb., 3 : 6 33.
49. Reece, K. S. Stokes, N. A. 2003. Molecular analysis of a haplosporidian parasite from cultured New Zealand abalone Haliotis iris. Dis. Aquat. Org., 53 : 61 66.
50. Reece, K., Bushek, D., Hudson, K. Graves, J. 2001. Geographic distribution of Perkinsus marinus genetic strains along the Atlantic and Gulf coasts of the USA. Mar. Biol., 139 : 1047 1055.
51. Reece, K. S., Siddall, M. E., Stokes, N. A. Burreson, E. M. 2004. Molecular phylogeny of the phylum Haplosporidia based on two independent gene sequences. J. Parasitol., 90 : 1111 1122.
52. Robledo, J. A. F., Nunes, P. A., Cancela, M. L. Vasta, G. R. 2002. Development of an in vitro clonal culture and characterization of the rRNA gene cluster of Perkinsus atlanticus, a protistan parasite of the clam Tapes decussatus. J. Eukaryot. Microbiol., 49 : 414 422.
53. Ruano, F. Cachola, R. 1986. Outbreak of a severe epizootic of Perkinsus marinus (Levine - 78) at Ria de Faro clam's culture beds. 2nd International Colloquium on Pathology in Marine Aquaculture. Porto, Portugal. p. 41 42.
54. Rubini, S., Capellaro, H., Alessi, A., Badini, L., Campanini, A. Marzadori, F. 1996. Perkinsus sp. nei bivalve dal Reno a Cesenatico. Laguna, 1 : 25 31.
55. Sagristà, E., Durfort, M. Azevedo, C. 1996. Ultrastructural study of the parasite, Perkinsus atlanticus (Apicomplexa), on the clam Ruditapes philippinarum, in the Mediterranean. Sci. Mar., 60 : 283 288.
56. Santmartí, M. M., García Valero, J., Montes, J., Pech, A. Durfort, M. 1995. Seguimiento del protozoo Perkinsus sp. en las poblaciones de Tapes decussatus y Tapes semidecussatus del delta del Ebro. In : Castelló, F. Calderer, A. (ed.), Actas del V Congreso Nacional de Acuicultura, S. Carlos de la Rápita. Universidad de Barcelona, Barcelona, Spain. p. 260 265.
57. Schott, E. J., Pecher, W. T., Okafor, F. Vasta, G. R. 2003. The protistan parasite Perkinsus marinus is resistant to selected reactive oxygen species. Exp. Parasitol., 105 : 232 240.
58. Soniat, T. M. 1996. Epizootiology of Perkinsus marinus disease of eastern oysters in the Gulf of Mexico. J. Shellfish Res., 15 : 35 43.
59. Sunila, I., Hamilton, R. M. Dungan, C. F. 2001. Ultrastructural characteristics of the in vitro cell cycle of the protozoan pathogen of oysters, Perkinsus marinus. J. Eukaryot. Microbiol., 48 : 348 361.
60. * and other methods). Version 10. Sinauer Associates, Sunderland, MA.
61. Thompson, J. D., Higgins, D. G. Gibson, T. J. 1994. Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res., 22 : 4673 4680.
62. Villalba, A., Reece, K. S., Ordas, M. C., Casas, S. M. Figueras, A. 2004. Perkinsosis in molluscs: a review. Aquat. Living Resour., 17 : 411 432.
63. Volety, A. K. Chu, F. L. E. 1997. Acid phosphatase activity in Perkinsus marinus, the protistan parasite of the American oyster, Crassostrea virginica. J. Parasitol., 83 : 1093 1098.
64. Wright, A. C., Ahmed, H., Gauthier, J. D., Silva, A. M. Vasta, G. R. 2002. cDNA cloning and characterization of two iron superoxide dismutases from the oyster parasite Perkinsus marinus. Mol. Biochem. Parasitol., 123 : 73 77.