Identification of the harmful dinoflagellate Vulcanodinium rugosum recovered from a ballast tank of a globally traveled ship in Port Tampa Bay, Florida, USA

Abstract The transport of microalgae in the ballast tanks of shipping vessels is of global concern because many algal species can survive in ballast tanks as nonmotile or cyst stages increasing the likelihood of introductions into foreign ports. In 2004 a peridinoid dinoflagellate was collected and isolated from ballast residuals of the merchant vessel Southern Fighter in Port Tampa Bay, Florida, USA. Light and electron microscopy of the motile and nonmotile peridinoid cells indicated the dinoflagellate was Vulcanodinium rugosum . Bayesian and maximum-parsimony (MP) phylogenetic analyses of V. rugosum strain CCFWC516 showed that sequenced regions of its LSU and ITS matched those of V. rugosum strains from Japan but were divergent from those strains from New Zealand and France. LC–MS analyses indicated that strain CCFWC516 did not produce the neurotoxin pinnatoxin, an ability that has been reported for other strains of this species. Analyses did show, however, that strain CCFWC516 did produce portimine, a cyclic imine produced by all other strains of this species. The M/V Southern Fighter visited numerous ports along the coast of Japan and the North Sea before sailing to Florida. The phylogenetic match to Japanese strains, as well as the ship's ballast exchange history, suggests that strain CCFWC516 originated from Japan. In light of the increase in global shipping traffic, increases in vessel size and capacity, and the increased connectivity between the Pacific Ocean and the Caribbean and Gulf of Mexico that will result from the widening of the Panama Canal, the introduction of nonnative, harmful algal species is an area of heightened concern and calls for increased vigilance.

[1]  J. Wolny,et al.  Methods for sampling and analysis of marine microalgae in ship ballast tanks: a case study from Tampa Bay, Florida, USA , 2011 .

[2]  Kirsty F. Smith,et al.  Production of pinnatoxins by a peridinoid dinoflagellate isolated from Northland, New Zealand , 2010 .

[3]  Jane M. Lewis Cyst-theca Relationships in Scrippsiella (Dinophyceae) and Related Orthoperidinioid Genera , 1991 .

[4]  Jennifer L. Molnar,et al.  Assessing the global threat of invasive species to marine biodiversity , 2008 .

[5]  D. Hutchins,et al.  Transport of the Harmful Bloom Alga Aureococcus anophagefferens by Oceangoing Ships and Coastal Boats , 2004, Applied and Environmental Microbiology.

[6]  Michael P. Cummings,et al.  PAUP* [Phylogenetic Analysis Using Parsimony (and Other Methods)] , 2004 .

[7]  M. Kawachi,et al.  Traditional Microalgae Isolation Techniques , 2005 .

[8]  K. Steidinger,et al.  IDENTIFICATION OF PFIESTERIA PISCICIDA (DINOPHYCEAE) AND PFIESTERIA‐LIKE ORGANISMS USING INTERNAL TRANSCRIBED SPACER‐SPECIFIC PCR ASSAYS 1 , 2003 .

[9]  A. Wilkins,et al.  Portimine: a bioactive metabolite from the benthic dinoflagellate Vulcanodinium rugosum , 2013 .

[10]  A. Hricko,et al.  Progress and Pollution: Port Cities Prepare for the Panama Canal Expansion , 2012, Environmental health perspectives.

[11]  N. Zeng,et al.  The first report of Vulcanodinium rugosum (Dinophyceae) from the South China Sea with a focus on the life cycle , 2012 .

[12]  C. Bolch The use of sodium polytungstate for the separation and concentration of living dinoflagellate cysts from marine sediments , 1997 .

[13]  E. R. Cox,et al.  Thecal plate tabulation and variation in Peridinium balticum (Pyrrhophyta : Peridiniales) , 1985 .

[14]  C. Miles,et al.  Pinnatoxins and spirolides in Norwegian blue mussels and seawater. , 2011, Toxicon : official journal of the International Society on Toxinology.

[15]  G. Procaccini,et al.  Intraspecific diversity in Scrippsiella trochoidea (Dinopbyceae): evidence for cryptic species , 2003 .

[16]  D. Hernández-Becerril,et al.  Morphology and life stages of the potentially pinnatoxin-producing thecate dinoflagellate Vulcanodinium rugosum from the tropical Mexican Pacific , 2013 .

[17]  D. Anderson,et al.  Species boundaries and global biogeography of the Alexandrium tamarense complex (Dinophyceae) 1 , 2007 .

[18]  E. Nezan,et al.  Vulcanodinium rugosum gen. et sp. nov. (Dinophyceae), un Nouveau Dinoflagellé Marin de la Côte Méditerranéenne Française , 2011 .

[19]  Michael T. Gastner,et al.  The complex network of global cargo ship movements , 2010, Journal of The Royal Society Interface.

[20]  Gustaaf M. Hallegraeff,et al.  Transport of toxic dinoflagellate cysts via ships' ballast water☆ , 1991 .

[21]  R. Kudela,et al.  Phylogenetic Relationships of Yessotoxin-Producing Dinoflagellates, Based on the Large Subunit and Internal Transcribed Spacer Ribosomal DNA Domains , 2008, Applied and Environmental Microbiology.

[22]  James T. Carlton,et al.  Shipping Study: The Role of Shipping in the Introduction of Non-indigenous Aquatic Organisms to the Coastal Waters of the United States (Other than the Great Lakes) and an Analysis of Control Options. , 1995 .

[23]  P. Arctander,et al.  Simplicity-correlated size growth of the nuclear 28S ribosomal RNA D3 expansion segment in the crustacean order isopoda , 1996, Journal of Molecular Evolution.

[24]  L. Crowder,et al.  An Overlooked Scale of Global Transport: Phytoplankton Species Richness in Ships' Ballast Water , 2000, Biological Invasions.

[25]  P. McCarron,et al.  Identification of pinnatoxins and discovery of their fatty acid ester metabolites in mussels ( Mytilus edulis ) from eastern Canada. , 2012, Journal of agricultural and food chemistry.

[26]  Gustaaf M. Hallegraeff,et al.  Transport of toxic dinoflagellates via ships ballast water: bioeconomic risk assessment and efficacy of possible ballast water management strategies , 1998 .

[27]  E. Bresnan,et al.  Distribution, diversity and toxin composition of the genus Alexandrium (Dinophyceae) in Scottish waters , 2010 .

[28]  E. Grosholz,et al.  Global Invasions of Marine and Estuarine Habitats by Non-Indigenous Species: Mechanisms, Extent, and Consequences' , 1997 .

[29]  John P. Huelsenbeck,et al.  MrBayes 3: Bayesian phylogenetic inference under mixed models , 2003, Bioinform..

[30]  C. Bolch,et al.  A review of the molecular evidence for ballast water introduction of the toxic dinoflagellates Gymnodinium catenatum and the Alexandrium “tamarensis complex” to Australasia , 2007 .

[31]  G. Hallegraeff,et al.  Dinoflagellate Vulcanodinium rugosum identified as the causative organism of pinnatoxins in Australia, New Zealand and Japan , 2011 .

[32]  I. Davidson,et al.  The biology of ballast water 25 years later , 2011, Biological Invasions.

[33]  Kirsty F. Smith,et al.  A dinoflagellate producer of pinnatoxin G, isolated from sub-tropical Japanese waters , 2011 .

[34]  John P. Huelsenbeck,et al.  MRBAYES: Bayesian inference of phylogenetic trees , 2001, Bioinform..

[35]  G. Hallegraeff,et al.  Reproductive compatibility among four global populations of the toxic dinoflagellate Gymnodinium catenatum (Dinophyceae) , 2001 .

[36]  D. Anderson,et al.  IDENTIFICATION OF GROUP‐ AND STRAIN‐SPECIFIC GENETIC MARKERS FOR GLOBALLY DISTRIBUTED ALEXANDRIUM (DINOPHYCEAE). II. SEQUENCE ANALYSIS OF A FRAGMENT OF THE LSU rRNA GENE 1 , 1994 .

[38]  J. Huelsenbeck,et al.  MRBAYES : Bayesian inference of phylogeny , 2001 .

[39]  C. Miles,et al.  Pinnatoxin G is responsible for atypical toxicity in mussels (Mytilus galloprovincialis) and clams (Venerupis decussata) from Ingril, a French Mediterranean lagoon. , 2013, Toxicon : official journal of the International Society on Toxinology.

[40]  David F. Reid,et al.  Current state of understanding about the effectiveness of ballest water exchange (BWE) in reducing aquatic nonindigenous species (ANS) introductions to the Great Lakes Basin and Chesapeake Bay, USA : synthesis and analysis of existing information , 2007 .

[41]  G. Hallegraeff,et al.  Production of pinnatoxins E, F and G by scrippsielloid dinoflagellates isolated from Franklin Harbour, South Australia , 2011 .