Marine Biology

, Volume 125, Issue 2, pp 421–426 | Cite as

Fragile algae: axenic culture of field-collected samples of Heterosigma carterae

  • L. Connell
  • R. A. Cattolico


Twelve cultures of the naturally wall-less, unicellular chromophyte Heterosigma carerae originating from North America and Japan were analyzed. Several of these had been recovered from blooms that were toxic to fish. Algal cultures were concentrated by centrifugation onto a Percoll pad. The cells were selectively treated with a cascade of antibiotics, including ampicillin, kanamycin, neomycin, nitrofuratoin, penicllin-G, rifampin, and streptomycin. Each isolate was treated with a specific subset of the antibiotics to attain axenicity. Doubling times of the individual isolates varied from 1.8 to 2.9 d, and were not influenced by previous antibiotic treatment. The studies were conducted from 1991–1994 at the University of Washington, Seattle, Washington, USA.


Japan Centrifugation Streptomycin North America Ampicillin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Becker EW (1994) Microalgae biotechnology and microbiology. Cambridge, Cambridge University PressGoogle Scholar
  2. Bein SJ (1954) A study of certain chromogenic bacteria isolated from “red tide” water with a description of a new species. Bull mar Sci Gulf Caribb 4: 110–119Google Scholar
  3. Boustead NC, Chang FH, Pridmore R, Todd P (1989) Big Glory Bay algal bloom identified. Freshwat Catch 39: 3–4Google Scholar
  4. Cattolico RA, Boothroyd JC, Gibbs SP (1976) Synchronous growth and plastid replication in the naturally wall-less alga Olisthodiscus luteus. P1 Physiol 57: 497–503Google Scholar
  5. Conrad W, Kufferath H (1954) Recherches sur les eau saumatres des environs de Lilloo. Perti II. Descriptive. Mém Inst r Sci nat Belg 127: 1–346Google Scholar
  6. Cottrell MT, Suttle CA (1993) Production of axenic cultures of Micromonas pusilla (Prasinophyceae) using antibiotics. J Phycol 29: 385–387Google Scholar
  7. Difco (1985) Difco Manual. Difco Laboratories, Detroit, Michigan, SAGoogle Scholar
  8. Dimanlig MLV, Taylor FJR (1985) Extracellular bacteria and toxin production in Protogonyaulax species. In: Anderson DM, White AW, Baden DG (eds) Third International Conference on Toxic Dinoflagellates. Elsevier, Amsterdam, pp 103–108Google Scholar
  9. Diven CL, Schones HK (1982) Production of axenic Gonyaulax cultures by treatment with antibiotics. Appl envirl Microbiol 44: 250–254Google Scholar
  10. Droop MR (1954) A note on the isolation of small marine algae and flagellates for pure cultures. Mar biol Ass UK 33: 511–514Google Scholar
  11. Droop MR (1967) A procedure for routine purification of algal cultures with antibiotics. Br phycol Bull 3: 295–297Google Scholar
  12. Figueiras FG, Niell FX (1987) Distribución estacional y espacial del fitaplancton en la ría de Pontevedra (NO de España) Investigación pesq 51: 293–320Google Scholar
  13. Haight R, Taylor FJR (1990) Distribution of potentially harmful phytoplankton species in the Northern Strait of Georgia, British Columbia. Can J Fish aquat Sciences 47: 2239–2350Google Scholar
  14. Hara Y, Inouye I, Chihara M (1985) Morphology and ultrastructure of Olisthodiscus luteus (Raphidophycear) with special reference to the taxonomy. Bot Mag, Tokyo 98: 251–262Google Scholar
  15. Harrel L (1990) Report on the red tide kill at the Manchester WA. National Marine Fisheries Service, Research Station, Manchester, WashingtonGoogle Scholar
  16. Hellegraeff CM (1993) A review of harmful algal blooms and their apparent global increase. Phycologia 32: 79–99Google Scholar
  17. Hobbie JE, Daley R, Jasper S (1977) Use of Nuclepore filters for counting bacteria by fluorescence microscopy. Appl envirl. Microbiol 33: 1225–1228Google Scholar
  18. Honjo T (1991) Overview on bloom dynamics and physiological ecology of Heterosigma akashiwo. In: Smayda TJ, Shimizu Y (eds) Fifth International Conference on Toxic Marine Phytoplankton. Elsevier, Amsterdam, pp 33–42Google Scholar
  19. Kohata K, Watanabe M (1989) Diel changes in the composition of photosynthetic pigments in Chattonella antiqua and Heterosigma akashiwo (Raphidophyceae) Okaichi A, Anderson DM, Nemoto M (eds) Red tides: biology, environmental science and toxicology. Elsevier, Amsterdam, pp 329–332Google Scholar
  20. Lackney JB, Clendenning KA (1965) Ecology of the microbiota of San Diego Bay, California. Trans S Diego Soc nat Hist 14: 9–40Google Scholar
  21. Lackney JB, Lackney EW (1963) Microscopic algae and protozoa in the waters near Plymouth in August 1962. J mar biol Ass UK 43: 797–805Google Scholar
  22. Lembeye G, Campodonico I (1984) First recorded bloom of the dinoflagellate Prorocentrum micans Enr. in South-central Chile. Botanica mar 27: 491–493Google Scholar
  23. McCracken R (1989) Purifying algal cultures—a review of chemical methods. Proc Trans Nova Scotian Inst Sci 38: 145–168Google Scholar
  24. McIntosh L, Cattolico RA (1978) Preservation of algal and higher plant ribosomal RNA integrity during extraction and electrophoretic quantitation. Analyt Biochem 91: 600–612Google Scholar
  25. Myers SP, Barlow MH (1959) Studies of Flavobacterium piscicida. I. Growth, toxicity and ecological considerations. J Bact 78: 225–230Google Scholar
  26. Parra OO, Rivera PR, Floyd GL, Wilcox LW (1991) Cultivo, morfología, ultraestructura y taxonomia de un fitoflagelado asociado a mareas rojas en Chile: Heterosigma akashiwo (Hada) Hada. Gayana, Chile (Bot) 48: 101–110Google Scholar
  27. Porter KG, Beig YS (1980) The use of DAPI for identifying and counting aquatic microflora. Limnol Oceanogr 25: 943–948Google Scholar
  28. Pratt DM (1966) Competition between Skeletonema costatum and Olisthodiscus luteus in Narragansett Bay and in culture. Limnol Oceanogr 11: 447–455Google Scholar
  29. Price CA, Roardon EM, Guillard RR (1978) Collection of dinoflagellates and other marine microalgae by centrifugation in density gradients of a modified silica sol. Limnol Oceanogr 23: 548–553Google Scholar
  30. Qi Y, Zhang Z, Hong Y, Lu S, Zhu C, Li Y (1991) Occurrence of red tides on the coasts of China. In: Smayda TJ, Shimizu Y (eds) Fifth International Conference on Toxic Marine Phytoplankton. Elsevier, Amsterdam, pp 43–46Google Scholar
  31. Reith ME, Cattolico RA (1985) In vitro chloroplast protein synthesis by the chromophytic alga Olisthodiscus luteus. P1 Physiol 24: 2550–2556Google Scholar
  32. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: A laboratory manual. Cold Spring Harbor, New York, Cold Spring Harbor PressGoogle Scholar
  33. Sherman F, Fink GR, Lawrence C (1979) Methods in yeast genetics. Cold Spring Harbor Press, Cold Spring Harbor, New YorkGoogle Scholar
  34. Shilo M, Asehner M (1953) Factors governing the toxicity of cultures containing the phytoflagellate Prymnesium parvum Carter. J gen Microbiol 8: 333–343Google Scholar
  35. Shumway SE (1990) A review of the effects of algal blooms on shellfish and aquaculture. J Wld Aquacult Soc 21: 65–104Google Scholar
  36. Sieburth JM, Keller MD (1989) Methylaminotrophic bacteria in xenic nanoalgal culture: Incidence, significance, and role of methylated algal osmoprotectants. Biol Oceanogr 6: 383–395Google Scholar
  37. Silva ES (1981) Relationship between dinoflagellates and intracellular bacteria. Proc 10th int Seaweed Symp 269-288 [In: Hoppe HA, Levring T, Tanaka Y (eds) W. de Gruyter & Co., Goteberg, Sweden]Google Scholar
  38. Takahashi M, Fukazawa M (1982) A mechanism of ”red-tide” formation. II. Effect of selected nutrient stimulation on the growth of different phytoplankton species in natural water. Mar Biol 70: 267–273Google Scholar
  39. Taniguchi A, Takeda Y (1988) Feeding rate and behavior of the tintinnid ciliate Favella taraikaensis observed with a high speed VTR system. Mar Microb Fd Webs 3: 21–34Google Scholar
  40. Taylor FJR (1990) Red tides, brown tides and other harmful algal blooms: the view into the 1990's. In: Graneli E, Sundstrom L, Edler L, Anderson DM (eds). Toxic marine phytoplankton. Elsevier, New York, pp 527–533Google Scholar
  41. Taylor FJR (1991) Current problems with harmful phytoplankton blooms in British Columbia waters. In: Smayda TJ, Shimizu Y (eds) Fifth International Conference on Toxic Marine Phytoplankton. Elsevier, Amsterdam, pp 699–704Google Scholar
  42. Taylor FJR (1992) The taxonomy of harmful marine phytoplankton. Giorn bot ital 126: 209–219Google Scholar
  43. Tomas CR (1980) Olisthodiscus luteus (Chrysophyceae). V. Its occurrence, abundance and dynamics in Narragansett Bay, Rhode Island. J Phycol 16: 157–166Google Scholar
  44. Tseng CK, Zhou MJ, Zou JZ (1991) Toxic phytoplankton studies in China. In: Smayda TJ, Shimizu Y (eds) Fifth International Conference on Toxic Marine Phytoplankton. Elsevier, Amsterdam, pp 347–352Google Scholar
  45. Uye S, Takamatsu K (1990) Feeding interactions between planktonic copepods and red tide flagellates from Japanese coastal waters. Mar Ecol Prog Ser 59: 97–107Google Scholar
  46. Wilson DP (1981) An experimental search for phytoplanktonic algae producing external metabolites which condition natural sea waters. J mar biol Ass UK 61: 585–607Google Scholar
  47. Yamaoka Y, Takimura O, Fuse H (1987) HPLC analysis of carbohydrates in two red tide phytoplanktons (Chattonella antiqua and Heterosigma akashiwo). J agric Chem Soc Japan 61: 23–28Google Scholar
  48. Yokote M, Honjo T, Asakawa M (1985) Histochemical demonstration of a glycocalyx on the cell surface of Heterosigma akashiwo. Mar Biol 88: 295–299Google Scholar
  49. Yoon YH, Rao HK, Kim YG (1991) Red tide organisms in the coastal waters of Chenju Island, southern Korea. Bull mar Res Inst, Chenju natn Univ 15: 1–14Google Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • L. Connell
    • 1
  • R. A. Cattolico
    • 2
  1. 1.Department of BotanyUniversity of WashingtonSeattleUSA
  2. 2.Department of OceanographyUniversity of WashingtonSeattleUSA

Personalised recommendations