Characterization of Dinophysis acuminata from the Yellow Sea, China, and its response to different temperatures and Mesodinium prey

Han Gao, Xinlong An, Lei Liu, Ke Zhang, Daoqiong Zheng, Mengmeng Tong

Light micrographs of cultured cells Dinophysis acuminata DAYS01, from Xiaoping Island, the Yellow Sea, China

Paper category: Original research paper
Corresponding author: Mengmeng Tong (
DOI: 10.1515/ohs-2017-0043
Received: March 7, 2017
Accepted: May 23, 2017
Full text: here

Citation (APA style):


Dinophysis species are distributed worldwide and cause diarrheic shellfish poisoning (DSP). This paper documents the first successful culture of a Dinophysis (DAYS01) strain taken off Xiaoping Island, the Yellow Sea, China. The strain was identified as Dinophysis acuminata Claparède & Lachmann by morphological and phylogenetic analysis. The effects of temperature and different Mesodinium prey on the physiological and toxigenic characteristics of D. acuminata DAYS01 were also investigated. The results showed that the toxin concentration (toxin amount per ml of culture) increased with an increase in cell densities. Okadaic acid (OA), dinophysistoxin 1 (DTX1) and pectenotoxin 2 (PTX2) were the major toxin components of DAYS01, with the highest recorded toxin content of 5.66, 0.56 and 192.87 ng ml-1 culture medium, respectively. The prey type significantly influenced the growth yield in Dinophysis cultures. There was an effect of prey on the total toxin content, suggesting that the origin or strain of ciliate directly impacted the D. acuminata blooms and the overall toxin concentration in the system.


Alves-de-Souza, C., Varela, D., Contreras, C., de La Iglesia, P., Fernández, P. et al. (2013). Seasonal variability of Dinophysis spp. and Protoceratium reticulatum associated to lipophilic shellfish toxins in a strongly stratified Chilean fjord. Deep-Sea Res. II: Topical Studies in Oceanography 101: 152-162.

Blanco, J., Alvarez, G. & Uribe, E. (2007). Identification of pectenotoxins in plankton, filter feeders, and isolated cells of a Dinophysis acuminata with an atypical toxin profile, from Chile. Toxicon 49(5): 710-716.

Chen, G.W. (1988). Taxonomic studies on three genera of dinophysiaceae in South China Sea. Oceanol. Limnol. Sinica 19(3): 238-248. (In Chinese).

Deeds, J.R., Wiles, K., Heideman, G.B., White, K.D. & Abraham, A. (2010). First US report of shellfish harvesting closures due to confirmed okadaic acid in Texas Gulf coast oysters. Toxicon 55(6): 1138-1146.

Edvardsen, B., Shalchian-Tabrizi, K., Jakobsen, K.S., Medlin, L.K., Dahl, E. et al. (2003). Genetic variability and molecular phylogeny of Dinophysis species (Dinophyceae) from Norwegian waters inferred from single cell analyses of rDNA. J. Phycol. 39(2): 395-408.

Fabro, E., Almandoz, G.O., Ferrario, M., Tillmann, U., Cembella, A. et al. (2016). Distribution of Dinophysis species and their association with lipophilic phycotoxins in plankton from the Argentine Sea. Harmful Algae 59: 31-41.

FAO. (2004). Marine Biotoxins. Food and Agriculture Organization of the United Nations, Rome.

Fernandez, M.L., Reguera, B., Gonzalez-Gil, S. & Miguez, A. (2006). Pectenotoxin-2 in single-cell isolates of Dinophysis caudata and Dinophysis acuta from the Galician Rias (NW Spain). Toxicon 48(5): 477-490.

Fux, E., Smith, J.L., Tong, M.M., Guzman, L. & Anderson D.M. (2011). Toxin profiles of five geographical isolates of Dinophysis spp. from North and South America. Toxicon 57(2): 275-287.

Guillard, R.R.L. (1973). Division rates. In J.R. Stein (Ed.), Handbook of Phycological Methods; Culture Methods and Growth Measurements (pp. 289-312). Cambridge University Press, Cambridge.

Guillou, L., Nezan, E. & Barbier, G. (2002). Genetic diversity of Dinophysis spp., AY040569-85, EMBL database,

Hackett, J.D., Tong, M.M., Kulis, D.M., Fux, E., Hess, P. et al. (2009). DSP toxin production de novo in cultures of Dinophysis acuminata (Dinophyceae) from North America. Harmful Algae 8(6): 873-879.

Hallegraeff, G.M. & Lucas, I.A.N. (1988). The Marine Dinoflagellate Genus Dinophysis (Dinophyceae) – photosynthetic, neritic and non-photosynthetic, oceanic species. Phycologia 27(1): 25-42.

Hattenrath-Lehmann, T.K. & Gobler, C.J. (2015). The contribution of inorganic and organic nutrients to the growth of a North American isolate of the mixotrophic dinoflagellate, Dinophysis acuminata. Limnol. Oceanogr. 60(5): 1588-1603.

Hattenrath-Lehmann, T.K., Marcoval, M.A., Mittlesdorf, H., Goleski, J.A., Wang, Z. et al. (2015). Nitrogenous nutrients promote the growth and toxicity of Dinophysis acuminata during estuarine bloom events. PLoS One 10(4): e0124148.

Holmes, M.J., Teo, S.L.M, Lee, F.C. & Khoo, H.W. (1999). Persistent low concentrations of diarrhetic shellfish toxins in green mussels Perna viridis from the Johor Strait, Singapore: first record of diarrhetic shellfish toxins from South-East Asia. Mar. Ecol. Prog. Ser. 181: 257-268.

Holmes, M.J., Teo, S.L.M. (2002). Toxic marine dinoflagellates in Singapore waters that cause seafood poisonings. Clinical and Experimental Pharmacology and Physiology 29(9): 829-836.

Jørgensen, K. & Andersen, P. (2007). Relation between the concentration of Dinophysis acuminata and diarrheic shellfish poisoning toxins in blue mussels (Mytilus edulis) during a toxic episode in the Limfjord (Denmark), 2006. J. Shellfish. Res. 26(4): 1081-1087.

Jaén, D., Mamán, L., Domínguez, R. & Martín, E. (2009). First Report of Dinophysis acuta in culture. Harmful Algae News 39: 1-2.

Jiang, T., Xu, Y.X, Li. Y., Qi, Y.Z., Jiang, T.J. et al. (2014). Dinophysis caudata generated lipophilic shellfish toxins in bivalves from the Nanji Islands, East China Sea. Chin. J. Oceanol. Limn. 32(1): 130-139.

Kamiyama, T. & Suzuki, T. (2009). Production of dinophysistoxin-1 and pectenotoxin-2 by a culture of Dinophysis acuminata (Dinophyceae). Harmful Algae 8(2): 312-317.

Kamiyama, T., Nagai, S., Suzuki, T. & Miyamura, K. (2010). Effect of temperature on production of okadaic acid, dinophysistoxin-1, and pectenotoxin-2 by Dinophysis acuminata in culture experiments. Aquat. Microb. Ecol. 60(2): 193-202.

Kim, S., Kang, Y.G., Kim, H.S., Yih, W., Coats, D.W. et al. (2008). Growth and grazing responses of the mixotrophic dinoflagellate Dinophysis acuminata as functions of light intensity and prey concentration. Aquat. Microb. Ecol. 51(3): 301-310.

Lee, J., Igarashi, T., Fraga, S., Dahl, E., Hovgaard, P. et al. (1989). Determination of diarrhetic shellfish toxins in various dinoflagellate species. J. Appl. Phycol. 1(2): 147-152.

Li, A., Ma, J., Cao, J. & McCarron, P. (2012). Toxins in mussels (Mytilus galloprovincialis) associated with diarrhetic shellfish poisoning episodes in China. Toxicon 60(3): 420-425.

Li, A.F., Sun, G., Qiu, J.B. & Fan, L. (2015). Lipophilic shellfish toxins in Dinophysis caudata picked cells and in shellfish from the East China Sea. Environ. Sci. Pollut. R. 22(4): 3116-3126.

Lin, S.J., Zhang, H.A., Spencer, D.F., Norman, J.E. & Gray, M.W. (2002). Widespread and extensive editing of mitochondrial mRNAs in dinoflagellates. J. Mol. Biol. 320(4): 727-739.

Luo, X. (2011). Population Dynamics and Toxin Production of Dinophysis Species in the Coastal Waters of Qingdao. Chinese Academy of Sciences, Beijing, China. pp. 61-78. (In Chinese).

MacKenzie, L., Beuzenberg, V., Holland, P., McNabb, P., Suzuki, T. et al. (2005). Pectenotoxin and okadaic acid-based toxin profiles in Dinophysis acuta and Dinophysis acuminata from New Zealand. Harmful Algae 4(1): 75-85.

Maestrini, Y.S. (1998). Blooms Dynamics and Ecophysiology of Dinophysis spp. In D.M. Anderson, A.D. Cembella & G.M. Hallegraeff (Eds.), Physiological Ecology of Harmful Algal blooms (pp. 243-265). Springer, Berlin, Germany.

Mafra, L.L., Tavares, C.P.D. & Schramm, M.A. (2014). Diarrheic toxins in field-sampled and cultivated Dinophysis spp. cells from southern Brazil. J. Appl. Phycol. 26(4): 1727-1739.

Marasigan, A.N., Sato, S., Fukuyo, Y. & Kodama, M. (2001). Accumulation of a high level of diarrhetic shellfish toxins in the green mussel Perna viridis during a bloom of Dinophysis caudata and Dinophysis miles in Sapian Bay, Panay Island, the Philippines. Fisheries. Sci. 67(5): 994-996.

Marín, I., Aguilera, A., Gonza´ lez-Gil, S., Reguera, B. & Abad, J.P. (2001). Genetic analysis of three species of Dinophysis causing diarrhetic shellfish outbreaks in Galicia (NW Spain). In G.M. Hallegraeff, C.J.S. Bolch, S.I. Blackburn & R.J. Lewis (Eds.), Harmful Algal Blooms 2000 (pp. 222-225). Intergovernmental Oceanographic Commission of UNESCO, Paris.

Nagai, S., Nitshitani, G., Tomaru, Y., Sakiyama S. & Kamiyama, T. (2008). Predation by the toxic dinoflagellate Dinophysis fortii on the ciliate Myrionecta rubra and observation of sequestration of ciliate chloroplasts. J. Phycol. 44(4): 909-922.

Nagai, S., Suzuki, T., Nishikawa, T. & Kamiyama, T. (2011). Differences in the Production and Excretion Kinetics of Okadaic Acid, Dinophysistoxin-1, and Pectenotoxin-2 between Cultures of Dinophysis Acuminata and Dinophysis Fortii Isolated from Western Japan. J. Phycol. 47(6): 1326-1337.

Nielsen, L., Krock, B. & Hansen, P. (2012). Effects of light and food availability on toxin production, growth and photosynthesis in Dinophysis acuminata. Marine Ecology Progress Series 471: 37-50.

Nielsen, L.T., Krock, B. & Hansen, P.J. (2013). Production and excretion of okadaic acid, pectenotoxin-2 and a novel dinophysistoxin from the DSP-causing marine dinoflagellate Dinophysis acuta – Effects of light, food availability and growth phase. Harmful Algae 23: 34-45.

Nishitani, G., Nagai, S., Sakiyama, S. & Kamiyama, T. (2008a). Successful cultivation of the toxic dinoflagellate Dinophysis caudata (Dinophyceae). Plankton and Benthos Research 3(2): 78-85.

Nishitani, G., Nagai, S., Takano, Y., Sakiyama, S., Baba, K. et al. (2008b). Growth characteristics and phylogenetic analysis of the marine dinoflagellate Dinophysis infundibulus (Dinophyceae). Aquat. Microb. Ecol. 52(3): 209-221.

Papaefthimiou, D., Aligizaki, K. & Nikolaidis, G. (2010). Exploring the identity of the Greek Dinophysis cf. acuminata. Harmful Algae 10(1): 1-8.

Park, M.G., Kim, S., Kim, H.S., Myung, G., Kang, Y.G. et al. (2006). First successful culture of the marine dinoflagellate Dinophysis acuminata. Aquat. Microb. Ecol. 45: 101-106.

Park, M.G., Kim, M., Kim, S. & Yih, W. (2010). Does Dinophysis caudata (Dinophyceae) have permanent plastids? J. Phycol. 46(2): 236-242.

Pizarro, G., Paz, B., Gonzalez-Gil, S., Franco, J.M. & Reguera, B. (2009). Seasonal variability of lipophilic toxins during a Dinophysis acuta bloom in Western Iberia: Differences between picked cells and plankton concentrates. Harmful Algae 8(6): 926-937.

Pizarro, G., Morono, A., Paz, B., Franco, J.M., Pazos, Y. et al. (2013). Evaluation of passive samplers as a monitoring tool for early warning of Dinophysis toxins in shellfish. Mar. Drugs. 11(10): 3823-3845.

Qiu, D., Huang, L., Liu, S. & Lin, S.J. (2011). Nuclear, mitochondrial and plastid gene phylogenies of Dinophysis miles (Dinophyceae): Evidence of variable types of chloroplasts. PloS One 6(12): e29398. DOI: 10.1371/journal.pone.0029398.

Raho, N., Pizarro, G., Escalera, L., Reguera, B. & Marín, I. (2008). Morphology, toxin composition and molecular analysis of Dinophysis ovum Schütt, a dinoflagellate of the “Dinophysis acuminata complex”. Harmful Algae 7(6): 839-848.

Raho, N., Rodríguez, F., Reguera, B. & Marín, I. (2013). Are the mitochondrial cox1 and cob genes suitable markers for species of Dinophysis Ehrenberg? Harmful Algae 28: 64-70.

Reguera, B., Velo-Suarez, L., Raine, R. & Park, M.G. (2012). Harmful Dinophysis species: A review. Harmful Algae 14: 87-106.

Reguera, B., Riobo, P., Rodriguez, F., Diaz, P.A., Pizarro, G. et al. (2014). Dinophysis toxins: causative organisms, distribution and fate in shellfish. Mar. Drugs. 12(1): 394-461.

Rial, P., Garrido, J.L., Jaen, D. & Rodriguez, F. (2013). Pigment composition in three Dinophysis species (Dinophyceae) and the associated cultures of Mesodinium rubrum and Teleaulax amphioxeia. J. Plankton. Res. 35(2): 433-437.

Rodríguez, F., Escalera, L., Reguera, B., Rial, P., Riobó, P. et al. (2012). Morphological variability, toxinology and genetics of the dinoflagellate Dinophysis tripos (Dinophysiaceae, Dinophysiales). Harmful Algae 13: 26-33.

Smith, J.L., Tong, M., Fux, E. & Anderson, D.M. (2012). Toxin production, retention, and extracellular release by Dinophysis acuminata during extended stationary phase and culture decline. Harmful Algae 19: 125-132.

Suzuki, T., Miyazono, A., Baba, K., Sugawara, R. & Kamiyama, T. (2009). LC-MS/MS analysis of okadaic acid analogues and other lipophilic toxins in single-cell isolates of several Dinophysis species collected in Hokkaido, Japan. Harmful Algae 8(2): 233-238.

Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution 30(12): 2725-2729.

Tong M.M., Kulis, D.M., Jiang, T.J., Qi, Y.Z., Zhou, Q.X. et al. (2010). Culture techniques and growth characteristics of Dinophysis acuminata and its prey. Chin. J. Oceanol. Limn. 28(6): 1230-1239.

Tong, M.M., Kulis, D.M., Fux, E., Smith, J.L., Hess, P. et al. (2011). The effects of growth phase and light intensity on toxin production by Dinophysis acuminata from the northeastern United States. Harmful Algae 10(3): 254-264.

Tong, M.M., Smith, J.L., Richlen, M., Steidinger, K.A., Kulis, D.M. et al. (2015a). Characterization and comparison of toxin-producing isolates of Dinophysis acuminata from new England and Canada. J. Phycol. 51(1): 66-81.

Tong, M.M., Smith, J.L., Kulis, D.M. & Anderson, D.M. (2015b). Role of dissolved nitrate and phosphate in isolates of Mesodinium rubrum and toxin-producing Dinophysis acuminata. Aquatic Microbial Ecology 75(2): 169-185.

Trainer, V., Moore, L., Bill, B., Adams, N., Harrington, N. et al. (2013). Diarrhetic shellfish toxins and other lipophilic toxins of human health concern in Washington state. Mar. Drugs. 11(6): 1815.

Wang, J.H., Qin, Y.T., Liu, C.C. & Xu, R. (2007). A background investigation of toxic algae and biotoxin in the Yangtze Estuary. Trans. Oceanol. Limnol. 53(1): 52-61.

Wotton, R.S. (1995). Temperature and lake-outlet communities. J. Therm. Biol. 20(1-2): 121-125.

Yasumoto, T., Oshima, Y. & Yamaguchi, M. (1978). Occurrence of a new type of shellfish poisoning in Tohoku district. B. Jpn. Soc. Sci. Fish. 44(11): 1249-1255.

Zhou, M., Li, J., Luckas, B., Yu, R., Yan, T. et al. (1999). A recent shellfish toxin investigation in China. Mar. Poll. Bull. 39(1): 331-334.


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