Abstract
Procedures have been developed for the extraction and high-performance liquid chromatography (HPLC) analysis of paralytic shellfish poisoning (PSP) toxins from Protogonyaulax spp. grown in batch culture. Using these procedures, the toxin content of two isolates of P. tamarensis (NEPCC 183 and 255) and one isolate of P. catenella (NEPCC 355) were examined. Total toxin and individual toxin concentrations were measured for each isolate during the exponential and stationary phases of growth in batch culture. The total toxicity of each isolate as measured by HPLC analysis was found to agree with toxicity as determined by the standard mouse bioassay. Two of the isolates (255 and 355) were found to be toxic and the third (183) was non-toxic. The toxic isolates (255 and 355) both showed higher average total PSP toxin content during the exponential phase (35 and 23 fmol toxin cell-1, respectively) than during the stationary phase (21 and 8 fmol toxin cell-1, respectively). These cultures differed dramatically in their toxin composition. P. tamarensis (255) contained a large proportion of the N(21) sulfo toxins (B1, B2, C1, C2) while P. catenella (355) contained primarily Gonyautoxins 1 through 4. The percent composition of individual toxins was found to be constant throughout the growth cycle for both toxic isolates, even though the total toxin concentration varied. Our results suggest that PSP toxin profiles might be useful as chemotaxonomic indicators.
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Alam, M. I., C. P. Hsu and Y. Shimizu: Comparison of toxins in three isolates of Gonyaulax tamarensis (Dinophyceae). J. Phycol. 15, 106–110 (1979)
Association of Official Agricultural Chemists (A.O.A.C.): Official methods of analysis, 12th ed., revised, Washington, D.C.: A.O.A.C. 1975
Bates, H. A., R. Kostriken and H. Rapoport: The occurrence of saxitoxin and other toxins in various dinoflagellates. Toxicon 16, 595–601 (1978)
Bates, H. A. and H. Rapoport: A chemical assay for saxitoxin, the paralytic shellfish poison. J. agric. Fd Chem. 23, 237–239 (1975)
Boyer, G.: Chemical investigations of the toxins produced by marine dinoflagellates, 226 pp. Ph.D. thesis, University of Wisconsin 1980
Boyer, G., J. J. Sullivan, R. J. Andersen, P. J. Harrison and F. J. R. Taylor: Toxin production in three isolates of Protogonyaulax sp. In: Toxic dinoflagellates, pp 281–286. Ed. by D. M. Anderson, A. W. White and D. G. Baden. New York: Elsevier/North Holland 1985
Fix-Wichmann, C., G. L. Boyer, C. L. Divan, E. J. Schantz and H. K. Schnoes: Neurotoxins of Gonyaulax excavata and Bay of Fundy scallops. Tetrahedron Lett. 22, 1941–1944 (1981)
Genenah, A. A. and Y. Shimizu: Specific toxicity of paralytic shellfish poisons. J. agric. Fd Chem. 29, 1289–1291 (1981)
Hall, S.: Toxins and toxicity of Protogonyaulax from the northeast Pacific, 196 pp. Ph.D. thesis, University of Alaska 1982
Harrison, P. J., R. E. Waters and F. J. R. Taylor: A broad spectrum artificial seawater medium for coastal and open ocean phytoplankton. J. Phycol. 16, 28–35 (1980)
Koehn, R. E., S. Hall, C. Fix-Wichmann, H. K. Schnoes and P. B. Reichardt: Dinoflagellate neurotoxins related to saxitoxin: structure and latent activity of toxins B1 and B2. Tetrahedron Lett. 23, 2247–2248 (1982)
Mickelson, C. and C. M. Yentsch: Toxicity and nucleic acid content of Gonyaulax excavata. Devs mar. Biol. 1, 131–134 (1979). (Proc. 2nd int. Conf. tox. Dinoflagellate Blooms, Key Biscayne, Fla, Oct. 31–Nov. 5, 1978. Ed. by D. L. Taylor and H. H. Seliger. New York: Elsevier/North Holland)
Oshima, Y., T. Hayakawa, M. Hashimoto, Y. Kotaki and T. Yasumoto. Classification of Protogonyaulax tamarensis from Northern Japan into three strains by toxin composition. Bull. Jap. Soc. scient. Fish. 48, 851–854 (1982)
Prakash, A.: Growth and toxicity of a marine dinoflagellate, Gonyaulax tamarensis. J. Fish. Res. Bd Can. 24, 1589–1606 (1967)
Prézelin, B. B.: Effects of light intensity on aging of the dinoflagellate Gonyaulax polyedra. Mar. Biol. 69, 129–135 (1982)
Proctor, N. H., S. L. Chan and A. J. Trevor: Production of saxitoxin by cultures of Gonyaulax catenella. Toxicon 13, 1–9 (1975)
Schmidt, R. J. and A. R. Loeblich, III: Distribution of paralytic shellfish poison among Pyrrophyta. J. mar. biol. Ass. U.K. 59, 479–487 (1979)
Shimizu, Y.: Dinoflagellate toxins: In: Marine natural products, chemical and biological perspectives, Vol. I pp 1–42. Ed. by P. Scheuer. New York: Academic Press 1978
Singh, H.T., Y. Oshima and T. Yasumoto: Growth and toxicity of Protogonyaulax tamarensis in axenic culture. Bull. Jap. Soc. scient. Fish. 48, 1341–1343 (1982)
Sullivan, J. J.: Paralytic shellfish poisoning — analytical and biochemical investigation, 260 pp. Ph.D. thesis. University of Washington 1982
Sullivan, J. J. and W. T. Iwaoka: High pressure liquid chromatographic determination of toxins associated with paralytic shellfish poisoning. J. Ass. off. analyt. Chem. 66, 297–303 (1983)
Sullivan, J. J. and M. M. Wekell: Determination of paralytic shellfish poisoning toxins by high pressure liquid chromatography. In: Seafood toxins, pp 197–205, Ed. by E. P. Ragelis. Washington, D.C.: American Chemical Society 1984
Sommer, H., W. F. Whedon, C. A. Kofoid and R. Stohler: Relation of paralytic shellfish poison to certain plankton organisms of the genus Gonyaulax. Archs Path. 24, 537–559 (1937)
Taylor, F. J. R.: Toxic dinoflagellates: taxonomic and biogeographic aspects with emphasis on Protogonyaulax. In: Seafood toxins, pp 77–97. Ed. by E. P. Ragelis. Washington, D.C.: American Chemical Society 1984
Trick, C. G., P. J. Harrison and R. J. Andersen: Extracellular secondary metabolite production by the marine dinoflagellate Prorocentrum minimum in culture. Can. J. Fish. aquat. Sciences 38, 864–867 (1981)
White, A. W. and L. Maranda: Paralytic toxins in the dinoflagellate Gonyaulax excavata and in shellfish. J. Fish. Res. Bd Can. 35, 397–402 (1978)
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Communicated by R. S. Carney, Baton Rouge
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Boyer, G.L., Sullivan, J.J., Andersen, R.J. et al. Use of high-performance liquid chromatography to investigate the production of paralytic shellfish toxins by Protogonyaulax spp. in culture. Marine Biology 93, 361–369 (1986). https://doi.org/10.1007/BF00401103
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DOI: https://doi.org/10.1007/BF00401103