Advertisement

Journal of Applied Phycology

, Volume 31, Issue 1, pp 519–532 | Cite as

Variations in morphology, growth, and toxicity among strains of the Prorocentrum lima species complex isolated from Cuba and Brazil

  • Angel R. Moreira-González
  • Luciano F. Fernandes
  • Hajime Uchida
  • Aya Uesugi
  • Toshiyuki Suzuki
  • Nicolas Chomérat
  • Gwenaël Bilien
  • Luiz L. MafraJrEmail author
Article

Abstract

Benthic dinoflagellates belonging to the genus Prorocentrum are known to produce diarrhetic toxins such as okadaic acid (OA) and its analogues, dinophysistoxins (DTXs), as well as prorocentrolides and other unidentified fast-acting toxins. The present study is a comparative analysis focused on the morphology, genetic, growth, toxin production, and toxicity by strains belonging to the Prorocentrum lima species complex, isolated from different regions along the western Atlantic coast. While cell dimensions (38–45 × 24–30 μm) and shape (ovoid) were similar between strains from Cuba and Recife (Northeast Brazil), cells of the strain from the estuarine complex of Paranaguá Bay (South Brazil) were shorter (36–41 × 25–28 μm) and oblong to ovate-oblong (elliptical) in shape. This latter strain exhibited similar LSU rDNA and identical ITS1-5.8S rDNA-ITS2 sequences to those of P. cf. lima “morphotype 5,” which is closely related to Prorocentrum caipirignum by LSU, but separated from it by ITS. This southern Brazilian strain attained the highest growth rate (0.34 ± 0.01 div day−1) and cell densities (11.2 × 105 cell mL−1) in batch culture. Intracellular OA concentrations were higher for the other two strains during the late exponential and stationary phase, but similar for all strains (9.50–10.06 pg cell−1) at the early exponential growth phase; consistently lower levels of DTX-1 were produced by two strains, except the one from Recife. Finally, live cells of the strains from Cuba and Recife were more toxic to A. salina metanauplii, whereas the southern Brazilian strain exhibited higher culture medium toxicity. The contrasts in growth and toxicity potential revealed for these morphologically and genetically distinct Prorocentrum strains might be relevant for the local management of diarrhetic poisoning outbreaks in shellfish harvesting and aquaculture sites, including in estuarine areas.

Keywords

Diarrhetic shellfish poisoning Harmful algae Laboratory culture Toxin production Toxicity to Artemia salina 

Notes

Funding information

This study was supported by the International Atomic Energy Agency (IAEA) through the Research Contract “Bentox Project,” IAEA RC# 18827. Authors are grateful to CAPES and CNPq (Brazil) for the Ph.D. scholarship awarded to A.R.M.G. through the co-funded PEC-PG Program, to Fundação Araucária (Brazil) for the grant awarded to L.L.M. Ligia F. G. da Luz (Universidade Federal do Paraná) assisted with graphic design, and Mutue T. Fuji (Instituto de Botânica) kindly invited one of the researchers to be part of the sampling campaign in Recife.

References

  1. Ajuzie CC (2007) Palatability and fatality of the dinoflagellate Prorocentrum lima to Artemia salina. J Appl Phycol 19:513–519CrossRefGoogle Scholar
  2. Ajuzie CC, Houvenaghel GT (2009) Preliminary survey of potentially harmful dinoflagellates in Nigeria’s coastal waters. Fottea 9:107–120CrossRefGoogle Scholar
  3. Aligizaki K, Nikolaidis G, Katikou P, Baxevanis AD, Abatzopoulos TJ (2009) Potentially toxic epiphytic Prorocentrum (Dinophyceae) species in Greek coastal waters. Harmful Algae 8:299–311CrossRefGoogle Scholar
  4. Ben-Gharbia H, Yahia OKD, Amzil Z, Chomérat N, Abadie E, Masseret E, Laabir M (2016) Toxicity and growth assessments of three thermophilic benthic dinoflagellates (Ostreopsis cf. ovata, Prorocentrum lima and Coolia monotis) developing in the southern mediterranean basin. Toxins 8:297–334CrossRefGoogle Scholar
  5. Bouaïcha N, Chézeau A, Turquet J, Quod JP, Puiseux-Dao S (2001) Morphological and toxicological variability of Prorocentrum lima clones isolated from four locations in the south-west Indian Ocean. Toxicon 39:1195–1202CrossRefGoogle Scholar
  6. Bravo I, Fernández ML, Ramilo I, Marínez A (2001) Toxin composition of the toxic dinoflagellate Prorocentrum lima isolated from different locations along the Galician coast (NW Spain). Toxicon 39:1537–1545CrossRefGoogle Scholar
  7. Brustolin MC, Thomas MC, Mafra LL, da Cunha Lana P (2014) Does Encope emarginata (Echinodermata: Echinoidea) affect spatial variation patterns of estuarine subtidal meiofauna and microphytobenthos? J Sea Res 91:70–78CrossRefGoogle Scholar
  8. dos Santos AA, Cocentino AMM, Reis TNV (2006) Macroalgas como indicadoras da qualidade ambiental da praia de Boa Viagem. Bol Téc Cient CEPENE, Tamandaré 14:25–33Google Scholar
  9. Foden J, Purdie DA, Morris S, Nascimento S (2005) Epiphytic abundance and toxicity of Prorocentrum lima populations in the Fleet Lagoon, UK. Harmful Algae 4:1063–1074CrossRefGoogle Scholar
  10. Gayoso A, Dover S, Morton S, Busman M, Moeller P, Fulco V, Maranda L (2002) Diarrhetic shellfish poisoning associated with Prorocentrum lima (Dinophyceae) in Patagonian gulfs (Argentina). J Shellfish Res 21:461–463Google Scholar
  11. Glibert PM, Burkholder JM, Kana TM (2012) Recent insights about relationships between nutrient availability, forms, and stoichiometry, and the distribution, ecophysiology, and food web effects of pelagic and benthic Prorocentrum species. Harmful Algae 14:231–259CrossRefGoogle Scholar
  12. Gouy M, Guindon S, Gascuel O (2010) SeaView version 4: a multiplatform graphical user interface for sequence alignment and phylogenetic tree building. Mol Biol Evol 27:221–224CrossRefGoogle Scholar
  13. Grzebyk D, Berland B, Thomassin BA, Bosi C, Arnoux A (1994) Ecology of ciguateric dinoflagellates in the coral reef complex of Mayotte Island (SW Indian Ocean). J Exp Mar Bio Ecol 178:51–66CrossRefGoogle Scholar
  14. Guillard RLR (1975) Culture of phytoplankton for feeding marine invertebrates. In: Smith WI, Chanley MH (eds) Culture of marine invertebrates. Plenum, NY, pp 29–59CrossRefGoogle Scholar
  15. Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O (2010) New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 59:307–321CrossRefGoogle Scholar
  16. Holmes MJ, Lee FC, Khoo HW, Teo SLM (2001) Production of 7-deoxy-okadaic acid by a New Caledonian strain of Prorocentrum lima (Dinophyceae). J Phycol 37:280–288CrossRefGoogle Scholar
  17. Hoppenrath M, Chomérat N, Horiguchi T, Schweikert M, Nagahama Y, Murray S (2013) Taxonomy and phylogeny of the benthic Prorocentrum species (Dinophyceae)—a proposal and review. Harmful Algae 27:1–28CrossRefGoogle Scholar
  18. Hu T, LeBlanc P, Burton IW, Walter JA, McCarron P, Melanson JE, Wright JL (2017) Sulfated diesters of okadaic acid and DTX-1: self-protective precursors of diarrhetic shellfish poisoning (DSP) toxins. Harmful Algae 63:85–93CrossRefGoogle Scholar
  19. Koike K, Sato S, Yamaji M, Nagahama Y, Kotaki Y, Ogata T, Kodama M (1998) Occurrence of okadaic acid producing Prorocentrum lima on the Sanriku coast, northern Japan. Toxicon 36:2039–2042CrossRefGoogle Scholar
  20. Lana PC, Marone E, Lopes RM, Machado EC (2001) The subtropical estuarine complex of Paranaguá Bay, Brazil. In: Selliger U, Kjerfve P (eds) Coastal marine ecosystems of Latin America. Springer, Berlin, pp 131–145CrossRefGoogle Scholar
  21. Lawrence JE, Bauder AG, Quilliam MA, Cembella AD (1998) Prorocentrum lima: a putative link to diarrhetic shellfish poisoning in Nova Scotia, Canada. In: Reguera B, Blanco J, Fernandez ML, Wyatt T (eds) Harmful microalgae. Xunta de Galicia and UNESCO, Santiago de Compostela, Spain, pp 78–79Google Scholar
  22. Laza-Martinez A, Orive E, Miguel I (2011) Morphological and genetic characterization of benthic dinoflagellates of the genera Coolia, Ostreopsis and Prorocentrum from the south-eastern Bay of Biscay. Eur J Phycol 46:45–65CrossRefGoogle Scholar
  23. Luo Z, Zhang H, Krock B, Lu S, Yang W, Gu H (2017) Morphology, molecular phylogeny and okadaic acid production of epibenthic Prorocentrum (Dinophyceae) species from the northern South China Sea. Algal Res 22:14–30CrossRefGoogle Scholar
  24. Mafra LL, Tavares CPS, Schramm MA (2014) Diarrheic toxins in field-sampled and cultivated Dinophysis spp. cells from southern Brazil. J Appl Phycol 26:1727–1739CrossRefGoogle Scholar
  25. Mafra LL, Ribas T, Alves TP, Proença LAO, Schramm MA, Uchida H, Suzuki T (2015) Differential okadaic acid accumulation and detoxification by oysters and mussels during natural and simulated Dinophysis blooms. Fish Sci 81:749–762CrossRefGoogle Scholar
  26. Marr JC, Jackson AE, McLachlan JL (1992) Occurrence of Prorocentrum lima, a DSP toxin-producing species from the Atlantic coast of Canada. J Appl Phycol 4:17–24CrossRefGoogle Scholar
  27. Moestrup Ø, Akselman R, Cronberg G, Elbraechter M, Fraga S, Halim Y, Hansen G, Hoppenrath M, Larsen J, Lundholm N, Nguyen LN, Zingone A (2009) IOC-UNESCO taxonomic reference list of harmful micro algae. Accessed at http://www.marinespecies. Org/hab (last accessed 20 January 2018)
  28. Morton SL, Tindall DR (1995) Morphological and biochemical variability of the toxic dinoflagellate Prorocentrum lima isolated from three locations at Heron Island, Australia. J Phycol 31:914–921CrossRefGoogle Scholar
  29. Nagahama Y, Murray S, Tomaru A, Fukuyo Y (2011) Species boundaries in the toxic dinoflagellate Prorocentrum lima (Dinophyceae, Prorocentrales), based on morphological and phylogenetic characters. J Phycol 47:178–189CrossRefGoogle Scholar
  30. Nascimento SM, Purdie DA, Morris S (2005) Morphology, toxin composition and pigment content of Prorocentrum lima strains isolated from a coastal lagoon in southern UK. Toxicon 45:633–649CrossRefGoogle Scholar
  31. Nascimento SM, Salgueiro F, Menezes M, de Andréa Oliveira F, Magalhães VCP, De Paula JC, Morris S (2016) Prorocentrum lima from the South Atlantic: morphological, molecular and toxicological characterization. Harmful Algae 57:39–48CrossRefGoogle Scholar
  32. Nascimento SM, Mendes MCQ, Menezes M, Rodríguez F, Alves-de-Souza C, Branco S, Morris S (2017) Morphology and phylogeny of Prorocentrum caipirignum sp. nov. (Dinophyceae), a new tropical toxic benthic dinoflagellate. Harmful Algae 70:73–89CrossRefGoogle Scholar
  33. Neves RA, Fernandes T, dos Santos LN, Nascimento SM (2017) Toxicity of benthic dinoflagellates on grazing, behavior and survival of the brine shrimp Artemia salina. PLoS One 12:e0175168CrossRefGoogle Scholar
  34. Nézan E, Tillmann U, Bilien G l, Boulben S, Chèze K, Zentz F, Salas R, Chomérat N (2012) Taxonomic revision of the dinoflagellate Amphidoma caudata: transfer to the genus Azadinium (Dinophyceae) and proposal of two varieties, based on morphological and molecular phylogenetic analyses. J Phycol 48:925–939CrossRefGoogle Scholar
  35. Nunn GB, Theisen BF, Christensen B, Arctander P (1996) Simplicity-correlated size growth of the nuclear 28S ribosomal RNA D3 expansion segment in the crustacean order Isopoda. J Mol Evol 42:211–223CrossRefGoogle Scholar
  36. Peraza R (2012) Composición y abundancia de dinoflagelados epibentónicos tecados potencialmente tóxicos de la Ensenada Guajimico, región Centro-Sur de Cuba. Bachelor Thesis. Universidad Central Marta Abreu de las VillasGoogle Scholar
  37. Posada D (2008) jModelTest: phylogenetic model averaging. Mol Biol Evol 25:1253–1256CrossRefGoogle Scholar
  38. Ronquist F, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19:1572–1574CrossRefGoogle Scholar
  39. Scholin C, Anderson DM (1994) Identification of group- and strain-specific genetic markers for globally distributed Alexandrium (Dinophyceae). I. RFLP analysis of SSU rRNA genes. J Phycol 30:744–754CrossRefGoogle Scholar
  40. Skinner MP, Lewis RJ, Morton S (2013) Ecology of the ciguatera causing dinoflagellates from the northern Great Barrier Reef: changes in community distribution and coastal eutrophication. Marine Poll Bull 77:210–219CrossRefGoogle Scholar
  41. Sobrinho BF, Mafra L (2016) Effects of irradiance and salinity on growth and toxin production by Prorocentrum lima. Proceedings 17th international conference on harmful algae, Oct 3–7 2016, Florianópolis, pp. 149 (Abstract)Google Scholar
  42. Takano H, Horiguchi T (2005) Acquiring scanning electron microscopical, light microscopical and multiple gene sequence data from a single dinoflagellate cell. J Phycol 42:251–256CrossRefGoogle Scholar
  43. Tindall DR, Morton SL (1998) Community dynamics and physiology of epiphytic/benthic dinoflagellates associated with ciguatera. NATO ASI Series G Ecological Sciences 41:293–314Google Scholar
  44. Tomas CR, Baden DG (1993) The influence of phosphorus source on the growth and cellular toxin content of the benthic dinoflagellate Prorocentrum lima. In: Toxic phytoplankton blooms in the sea. Proc. 5th lnt. Conf. on Toxic Marine Phytoplankton. Elsevier, Amsterdam pp. 565–570Google Scholar
  45. Valdiglesias V, Prego-Faraldo MV, Pásaro E, Méndez J, Laffon B (2013) Okadaic acid: more than a diarrheic toxin. Mar Drugs 11:4328–4349CrossRefGoogle Scholar
  46. Vanucci S, Guerrini F, Milandri A, Pistocchi R (2010) Effects of different levels of N- and P-deficiency on cell yield, okadaic acid, DTX-1, protein and carbohydrate dynamics in the benthic dinoflagellate Prorocentrum lima. Harmful Algae 9:590–599CrossRefGoogle Scholar
  47. Yoo JS (2004) Morphological variation of dinoflagellate Prorocentrum lima (Ehrenberg) Dodge with geographical difference. J Environ Biol 25:51–58PubMedGoogle Scholar
  48. Zhang H, Li Y, Cen J, Wang H, Cui L, Dong Y, Lu S (2015) Morphotypes of Prorocentrum lima (Dinophyceae) from Hainan Island, South China Sea: morphological and molecular characterization. Phycologia 54:503–516CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Centro de Estudos do MarUniversidade Federal do ParanáPontal do ParanáBrazil
  2. 2.Centro de Estudios Ambientales de Cienfuegos (CEAC)CienfuegosCuba
  3. 3.Departamento de BotânicaUniversidade Federal do ParanáCuritibaBrazil
  4. 4.National Research Institute of Fisheries ScienceYokohamaJapan
  5. 5.Ifremer, LER BO, Station de Biologie MarineConcarneauFrance

Personalised recommendations