Abstract
A strain of Alexandrium leei Balech that was isolated in October 2002 from Singapore coastal waters and identified by light and scanning electron microscopy and phylogenetic analysis using LSU rDNA sequences, is toxic to Asian sea bass fingerlings (Lates calcarifer Bloch). The ichthyotoxicity of the algal cells obtained by filtration (15 μm mesh net) and rinsed with sterile culture medium indicated that the toxicity of A. leei was probably not due to bacterial contamination, which was further supported by a negative correlation between the time to death of the fish and the dosage of algal cells applied. Paralytic shellfish toxins (PST) could not be detected indicating that PST was not the cause of fish mortality. Fish bioassays using frozen culture, heat-treated cultures, cell-free culture medium, and hexane, ethyl acetate, and water extracts of algal cells indicated that A. leei produces a heat-stable, polar ichthyotoxin(s) which can be released from the algal cells into the culture medium. Phylogenetic analysis based on the LSU rDNA sequences of A. leei confirmed its identification and indicated that the Singapore strain is more similar to isolates from Malaysia than to a geographically distant strain from Korea. This is the first evidence of icthyotoxin production by A. leei.
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References
Aldrich DV, Ray SM, Wilson WB (1967) Gonyaulax monilata: population growth and development of toxicity in culture. J Protozool 14:636–639
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Balech E (1985) The genus Alexandrium or Gonyaulax of the tamarensis group. In: Anderson DM, White AW, Baden DC (eds) Toxic dinoflagellates. Elsevier, Oxford, pp 33–38
Balech E (1995) The genus Alexandrium Halim. Sherkin Island Marine Station, Cork, p 151
Cembella AD, Lewis NI, Quilliam MA (2000) The marine dinoflagellate Alexandrium ostenfeldii (Dinophyceae) as the causative organism of spirolide shellfish toxins. Phycologia 39:67–74
Cembella AD, Quilliam MA, Lewis NI, Bauder AG, Dell’Aversano C, Thomas K, Jellett J, Cusack RR (2002) The toxigenic marine dinoflagellate Alexandrium tamarense as the probable cause of mortality of caged salmon in Nova Scotia. Harmful Algae 1:313–325
Chou R, Lee HB (1997) Commercial marine fish farming in Singapore. Aquac Res 28:767–776
Daugbjerg N, Hansen G, Larsen J, Moestrup Ø (2000) Phylogeny of some of the major genera of dinoflagellates based on ultra-structure and partial LSU rDNA sequence data, including the erection of three new genera of unarmoured dinoflagellates. Phycologia 39:302–317
Fritz L, Triemer RE (1985) A rapid simple technique utilizing Calcofluor White M2R for the visualization of dinoflagellate thecal plates. J Phycol 21:662–664
Fukuyo Y, Pholpunthin P, Yoshida K (1988) Protogonyaulax (Dinophyceae) in the Gulf of Thailand. Bull Plankton Soc Jpn 35:9–20
Gates JA, Wilson WB (1960) The toxicity of Gonyaulax monilata Howell to Mugil cephalus. Limnol Oceanogr 5:171–174
Hansen PJ, Cembella AD, Moestrup Ø (1992) The marine dinoflagellate Alexandrium ostenfeldii: paralytic shellfish toxin concentration, composition, and toxicity to a tintinnid ciliate. J Phycol 28(5):597–603
Holmes MJ, Lewis RJ, Gillespie NC (1990) Toxicity of Australian and French Polynesian strains of Gambierdiscus toxicus (Dinophyceae) grown in culture: characterization of a new type of maitotoxin. Toxicon 28:1159–1172
Holmes MJ (1998) Gambierdiscus yasumotoi sp. nov. (Dinophyceae), a toxic benthic dinoflagellate from southeastern Asia. J Phycol 34:661–668
Holmes MJ, Bolch CJS, Green DH, Cembella AD, Teo SLM (2002) Singapore isolates of the dinoflagellate Gymnodinium catenatum (Dinophyceae) produce a unique profile of paralytic shellfish poisoning toxins. J Phycol 38:96–106
Huelsenbeck JP, Ronquist F (2001) MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 17:754–755
Kim KY, Yoshida M, Kim CH (2005) Molecular phylogeny of three hitherto unreported Alexandrium species: Alexandrium hiranoi, Alexandrium leei and Alexandrium satoanum (Gonyaulacales, Dinophyceae) inferred from the 18S and 26S rDNA sequence data. Phycologia 44:361–368
Kodama M, Ogata T, Fukuyo Y, Igarashi T, Kamiya H, Matsuura F (1982) Comparison of toxicities of Protogonyaulax cells of various sizes. Bull Jpn Soc Sci Fish 48:567–574
Konovalova GV (1989) Phytoplankton blooms and red tides in the Far East coastal waters of the USSR. In: Okaichi T, Anderson DM, Nemoto T (eds) Red tides: biology, environmental science, and toxicology. Elsevier, Amsterdam, pp 97–100
Liang S, Qian HL, Qi YZ (2000) Problems on the red tide in coastal China sea. In: Ho KC, Wang ZD (eds) Prevention and management of harmful algal blooms in the South China Sea: proceedings of regional symposium on prevention management of harmful algal blooms in the South China Sea, 2000 HABSCS, pp 492
Matsuoka K, Iizuka H, Takayama H, Honjo T, Fukuyo Y, Ishimaru T (1989) Geographic distribution of Gymnodinium nagasakiense Takayama et Adachi around west Japan. In: Okaichi T, Anderson DM, Nemoto T (eds) Red tides: biology, environmental science, and toxicology. Elsevier, Amsterdam, pp 101–104
Nguyen-Ngoc L (2004) An autecological study of the potentially toxic dinoflagellate Alexandrium affine isolated from Vietnamese waters. Harmful Algae 3:117–129
Nylander JAA (2004) MrModeltest v2 (program distributed by the author). Evolutionary Biology Centre, Uppsala University
Ogata T, Kodama M (1986) Ichthyotoxicity found in cultured media of Protogonyaulax spp. Mar Biol 92:31–34
Posada D, Crandall KA (1998) Modeltest: testing the model of DNA substitution. Bioinformatics 14:817–818
Ronquist F, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19:1572–1574
Scholin CA, Villac MC, Buck KR, Krupp JM, Powers DA, Fryxell GA, Chavez EP (1994) Ribosomal DNA sequences discriminate among toxic and non-toxic Pseudonitzschia species. Natural Toxins 2:152–165
Steidinger KA, Tangen K (1997) Dinoflagellates. In: Tomas CR (ed) Identifying marine phytoplankton. Academic, New York, pp 387–589
Su HM, Liao IC, Chiang YM (1993) Mass mortality of prawn caused by Alexandrium tamarense blooming in a culture pond in southern Taiwan. In: Smayda TJ, Shimizu Y (eds) Toxic phytoplankton blooms in the Sea. Elsevier, Amsterdam, pp 329–333
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl Acids Res 25:4876–4882
Usup G, Leaw CP, Ahmad A, Lim PT (2002) Alexandrium (Dinophyceae) species in Malaysian waters. Harmful Algae 1:265–275
Acknowledgements
We thank Mr. Lim Huan Sein and Mr. Koh Chee Boon from the Marine Aquaculture Center, Agri-food and Veterinary Authority of Singapore for kindly providing the sea bass, Dr. Ying Han Gong for helping with toxin extraction, and Mr. Lim Poh Teen for useful suggestions. Ms. Alice Ilaya Gedaria helped maintain algal cultures. We thank Drs. Konda Reddy and Balasubramaniam Sivaloganathan for advice on the culture and physiological requirements of sea bass. This work was financially supported by NUS grants R-347-000-043-112 and R-154-000-108-112. The experiments presented in the study complied with the current laws of Singapore.
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Communicated by J.P. Grassle, New Brunswick
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Tang, Y.Z., Kong, L. & Holmes, M.J. Dinoflagellate Alexandrium leei (Dinophyceae) from Singapore coastal waters produces a water-soluble ichthyotoxin. Mar Biol 150, 541–549 (2007). https://doi.org/10.1007/s00227-006-0396-z
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DOI: https://doi.org/10.1007/s00227-006-0396-z