Scientia Marina, Vol 72, No 2 (2008)

Identification and characterisation of the dominant Pseudo-nitzschia species (Bacillariophyceae) along the NE Spanish coast (Catalonia, NW Mediterranean)

Sonia Quijano-Scheggia
Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar-CMIMA (CSIC), Spain

Esther Garcés
Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar-CMIMA (CSIC), Spain

Nagore Sampedro
Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar-CMIMA (CSIC), Spain

Kees van Lenning
Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar-CMIMA (CSIC), Spain

Eva Flo
Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar-CMIMA (CSIC), Spain

Karl Andree
Centre d’Aquicultura-IRTA, Spain

José-Manuel Fortuño
Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar-CMIMA (CSIC), Spain

Jordi Camp
Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar-CMIMA (CSIC), Spain


Potentially toxic species of the genus Pseudo-nitzschia were studied along the Spanish NW Mediterranean coast from January 2005 to May 2006. Observation in electron microscopy revealed the presence of Pseudo-nitzschia brasiliana, P. calliantha, P. delicatissima, P. fraudulenta, P. multistriata and P. pungens. Several strains were isolated from coastal waters and their clonal cultures were compared by combined techniques, including light and electron microscopy and internal transcribed spacer (ITS-1, 5.8S and ITS-2) rDNA sequencing. Cultured isolates were submitted to HPLC analysis of pigments to evaluate the possibility of taxonomic discriminations by means of a simple chemotaxonomic approach. The genus Pseudo-nitzschia showed high cell concentrations during most of the year, but the population initiated a sharp decline at all stations in the period between April and May. P. delicatissima with P. calliantha were found at the northern stations between February and April, and at the southern stations between August and November. P. brasiliana and P. multistriata were only occasionally detected in the southern region. None of the environmental variables considered was significant to explain the observed spatial and temporal distributions of Pseudo-nitzschia species in the area studied. Differences in the growth rate and cell yield of the species indicate that biotic factors may play a role in the observed distributional patterns.


diatoms; Pseudo-nitzschia; harmful algal blooms; autoecology; ITS morphology; pigments

Full Text:



Amato, A., W. Kooistra, J.H.L. Ghiron, D.G. Mann, T. Pröscholdand M. Montresor. – 2007. Reproductive isolation among sympatric cryptic species in marine diatoms. Protist, 158: 193-207. doi:10.1016/j.protis.2006.10.001

Amzil, Z., J. Fresnel, D. Le Gal and C. Billard. – 2001. Domoic acid accumulation in french sehllfish in relation to toxic species of Pseudo-nitzschia multiseries and P. pseudodelicatissima. Toxicon, 39: 1245-1251. doi:10.1016/S0041-0101(01)00096-4

Bates, S.S. – 2000. Domoic-acid-producing diatoms: another genus added! J. Phycol., 36: 978-983. doi:10.1046/j.1529-8817.2000.03661.x

Bates, S.S., C.J. Bird, A.S.W. de Freitas, R. Foxall, M. Gilgan, L.A. Hanic, G.R. Johnson, A.W. McCulloch, P. Odense, R. Pocklington, M.A. Quilliam, P.G. Sim, J.C. Smith, D.V. Subba Rao, E.C.D. Tood, J.A. Walter and J.L.C. Wright. – 1989. Pennate diatom Nitzschia pungens as the primary source of domoic acid, a toxin in shellfish from eastern prince Edward island, Canada. Can. J. Fish. Aquat. Sci., 46: 1203-1215.

Bates, S.S. and N.A. Davidovich. – 2002. Factors affecting the sexual reproduction of the diatoms, with emphasis on Pseudonitzschia spp. In: E. Gárces, A. Zingone, B. Dale, M. Montresor and B. Reguera (eds.), The harmful algal blooms, pp. 27-30. Luxembourg.

Bates, S.S. and V.L. Trainer. – 2006. The ecology of harmful diatoms. In: E. Granéli and J. Turner (eds.), Ecology of Harmful Algae, pp. 81-93. Springer-Verlag, Heidelberg.

Caropo, C., R. Congestri, l. Brancchini and P. Albertano. – 2005. On the presence of Pseudo-nitzschia calliantha Lundholm, Moestrup et Hasle and Pseudo-nitzschia delicatissima (Cleve) Heiden in the southern adriatic sea (Mediterranean sea, Italy). J. Plankton Res., 27: 763-774. doi:10.1093/plankt/fbi050

Clark, K.R. and R.M. Warwick. – 2001. Change in marine communities: an approach to statistical analysis and interpretation. PRIMER-E, Plymouth, U. K.

Coleman, A.W. – 2003. ITS2 is a double-edged tool for eukaryote evolutionary comparisons. Trends Genet.(Regul. ed.). 19: 370-375.

Drebes, G. – 1977. Sexuality. In: E.D. Werner (ed.), The Biology of Diatoms, pp. 250-283. Blackwell Scientific Publications, Oxford.

Furones, D., M. Vila, E. Garcés, N. Sampedro, L. Arín, M. Masó, J. Camp, K. van Lenning, S. Qujano-Scheggia, M. Delgado, M. Fernández, E. Mallat, E. Cañete, A. Caillaud and J. Diogene. – 2004. The Monitoring Programme for marine toxins and harmful phytoplankton in the Catalan coastline, North Western Mediterranean, Spain., Int. Conf. Molluscan Saf, Galway, Ireland.

Grasshoff, K., M. Ehrrardt and K. Kremling. – 1983. Methods of sea water analysis. Chemie, Germany.

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, New York.

Guillard, R.R.L. – 1975. Culture of phytoplankton for feeding marine invertebrates. In: W.L. Smith and M.H. Chanley (eds.), Culture of Marine Invertebrate Animals, pp. 29-60. Plenum Publishing Corp New York.

Guillard, R.R.L. and P.E. Hargraves. – 1993. Stichochrysis immobilis is a diatom, not a chrysophyte. Phycologia, 32: 234-236.

Hasle, G.R. – 1995. Pseudo-nitzschia pungens and P. multiseries (Bacillariophyceae): nomenclatural history, morphology, and distribution. J. Phycol., 31: 428-435. doi:10.1111/j.0022-3646.1995.00428.x

Hasle, G.R., C.B. Lange and E.E. Syvertsen. – 1996. A review of Pseudo-nitzschia, with special reference to the Skagerrak, North Atlantic, and adjacent waters. Helgol. Meeresunters., 50: 131-175. doi:10.1007/BF02367149

Hasle, G.R. and E.E. Syvertsen. – 1997. Marine diatoms. In: C.R. Tomas (ed.), Identifying marine phytoplankton, pp. 5-386. Academia Press, San Diego, California, USA, San Diego, California, USA.

Idescat. – 2006. Anuari estadístic de Catalunya. Institut d’Estadística de Catalunya, Barcelona.

Inès, S. and S.H. Asma. – 2006. First detection of toxic Pseudonitzschia calliantha in Bizerte Lagoon, Tunisia. Harmful Algae News, 30: 8-9.

Justic, D., N.N. Rabalais, R.E. Turner and Q. Dortch. – 1995. Changes in nutrient structure of river-dominated coastal waters: Stoichiometric nutrient balance and its consequences. Estuar. Coast Shelf. Sci., 40: 339-356. doi:10.1016/S0272-7714(05)80014-9

Kaczmarska, I., J.M. Ehrman, S.S. Bates, D.H. Green, C. Leger and J. Harris. – 2005. Diversity and distribution of epibiotic bacteria on Pseudo-nitzschia multiseries (Bacillariophyceae) in culture, and comparison with those on diatoms in native seawater. Harmful Algae, 4: 725-741. doi:10.1016/j.hal.2004.10.001

Kaniou-Grigoriadou, I., T. Mouratidou and P. Katikou. – 2005. Investigation on the presence of domoic acid in Greek shellfish. Harmful Algae, 4: 717-723. doi:10.1016/j.hal.2004.10.002

Lundholm, N., P. Andersen, K. Joergensen, B. Ring and A.D. Cembella. – 2005. Domoic acid in Danish blue mussels due to a bloom of Pseudo-nitzschia seriata. Harmful Algae News, 29: 6-9.

Lundholm, N., N. Daugbjerg and Ø. Moestrup. – 2002a. Phylogeny of the Bacillariaceae with emphasis on the genus Pseudonitzschia (Bacillariophyceae) based on partial LSU rDNA. Eur. J. Phycol., 37: 115-134. doi:10.1017/S096702620100347X

Lundholm, N., G.R. Hasle, G.A. Fryxell and P.E. Hargraves. – 2002b. Morphology, phylogeny and taxonomy of species within the Pseudo-nitzschia americana complex (Bacillariophyceae) with descriptions of two new species, Pseudo-nitzschia brasiliana and Pseudo-nitzschia linea. Phycologia, 41: 480-497.

Lundholm, N., Ø. Moestrup, G.R. Hasle and K. Hoef-Emden. – 2003. A study of the Pseudo-nitzschia pseudodelicatissima/cuspidate complex (Bacillariophyceae): what is P. pseudodelicatissima? J. Phycol., 39: 797-813. doi:10.1046/j.1529-8817.2003.02031.x

Mann, D.G. – 2002. Life cycles in diatoms. In: E. Garcés, A. Zingone, M. Montresor, B. Reguera and B. Dale, (eds.), LIFEHAB WORKSHOP: Life history of microalgal species causing harmful algal blooms. Commission of the european community,

Mann, D.G. and S.J.M. Droop. – 1996. 3. Biodiversity, biogeography and conservation of diatoms. Hydrobiologia, 336: 19-32.

Margalef, R. – 1969. Composición específica del fitoplancton de la costa catalano-levantina (Mediterráneo occidental) en 1962-1967. Invest. Pesq., 33: 345-380.

Massuti, M. and R. Margalef. – 1950. Introducción al estudio del plancton marino. Patronato Juan de la Cierva de Investigación Técnica, Barcelona.

Mos, L. – 2001. Domoic acid: a fascinating marine toxin. Environ. Toxicol. Pharmacol., 9: 79-85. doi:10.1016/S1382-6689(00)00065-X

Orsini, L., D. Sarno, G. Procaccini, R. Poletti, J. Dahlmann and M. Montresor. – 2002. Toxic Pseudo-nitzschia multistriata (Bacillariophyceae) from the Gulf of Naples: morphology, toxin analysis and phylogenetic relationships with other Pseudo-nitzschia species. Eur. J. Phycol., 37: 247-257. doi:10.1017/S0967026202003608

Orsini, L., G. Procaccini, D. Sarno and M. Montresor. – 2004. Multiple rDNA ITS-types within the diatom Pseudo-nitzschia delicatissima (Bacillariophyceae) and their relative abundance across a spring bloom in the Gulf of Naples. Mar. Ecol. Prog. Ser., 271: 87-89. doi:10.3354/meps271087

Penna, A., C. Ingarao, M. Ercolessi, M. Rocchi and N. Penna. – 2006. Potentially harmful microalgal distribution in an area of the NW Adriatic coastline: Sampling procedure and correlations with environmental factors. Estuar. Coast. Shelf. Sci., 70: 307-316. doi:10.1016/j.ecss.2006.06.023

Priisholm, K., Ø. Moestrup and N. Lundholm. – 2002. Taxonomic notes on the marine diatom genus Pseudo-nitzschia in the Andaman Sea near the island of Phuket, Thailand, with a description of Pseudo-nitzschia micropora sp nov. Diatom Res., 17: 153-175.

Quijano-Scheggia, S., E. Garcés, K. van Lenning, N. Sampedro and J. Camp. – 2005. First detection of diatom Pseudo-nitzschia brasiliana (non toxic) and its relative P. multistriata (presumably toxic) in the NW Mediterranean Sea. Harmful Algae News, 29: 5.

Quiroga, I. – 2006. Pseudo-nitzschia blooms in the Bay of Banyuls- sur-Mer, northwestern Mediterranean Sea. Diatom Res., 21: 91-104.

Round, F.E. – 1996. Fine detail for siliceous components of diatom cells. Beih. Nova Hedwig., 112: 201-213.

Round, F.E., R.M. Crawford and D.G. Mann. – 1990. The diatoms. Biology and morphology of the genera. Cambridge University Press, Cambridge.

Schaffer, P., C. Reeves, D.R. Casper and C.R. Davis. – 2006. Absence of neurotoxic effects in leopard sharks, Triakis semifasciata, following domoic acid exposure. Toxicon, 47: 747-752. doi:10.1016/j.toxicon.2006.01.030

Sierra-Beltran, A.P., R. Cortés-Altamirano, J.P. Gallo-Reynoso, S. Licea-Duran and J. Égido-Villarreal. – 2005. Is Pseudonitzschia pseudodelicatissima toxin the principal cause of sardines, dolphins, sea lions and pelican mortality in 2004 in Mexico? Harmful Algae News, 29: 6-8.

Sournia, A., M. Chretiennot-Dinet and M. Richter. – 1991. Marine phytoplankton:how many species in the world ocean? J. Plankton Res., 13: 1093-1099. doi:10.1093/plankt/13.5.1093

StatSoft, I. – 2003. STATISTICA (data analysis software system), version 6. StatSoft, Inc.

Stolte, W. and E. Garcés. – 2006. Ecological aspects of harmful algal in situ population growth rates. In: E. Granéli and J.T.

Turner (eds.), Ecology of Harmful Algae, pp. 139-152. Springer- Verlag Berlin, Berlin.

Throndsen, J. – 1995. Estimating cell numbers. In: G.M. Hallegraeff, D.M. Anderson and A.D. Cembella (eds.), Manual on Harmful Marine Microalgae, pp. 63-80. IOC Manuals and Guides UNESCO, Paris.

Trainer, V.L. and M. Suddleson. – 2005. Monitoring approaches for early warning of domoic acid events in Washington State. Oceanography, 18: 228-237.

Utermöhl, H. – 1931. Neue Wege in der quantitativen Erfassung des Planktons. Verh. int. Ver. Limnol., 5: 567-595.

Van Lenning, K., M. Latasa, M. Estrada, A.G. Sáez, L. Medlin, I. Probert, B. Véron and J. Young. – 2003. Pigment signatures and phylogenetic relationships of the pavlovophyceae (Haptophyta). J. Phycol., 39: 379-389.

Vila, M. – 2001. Harmful Algal Bloom in the Catalan coast (NW Mediterranean). PhD thesis. Univ. Barcelona.

Vila, M., J. Camp, E. Garcés, M. Maso and M. Delgado. – 2001. High resolution spatio-temporal detection of potentially harmful dinoflagellates in confined waters of NW Mediterranean. J. Plankton Res., 23: 497-514. doi:10.1093/plankt/23.5.497

Zapata, M., F. Rodríguez and J.L. Garrido. – 2000. Separation of chlorophylls and carotenoids from marine phytoplankton: a new HPLC method using a reversed phase C_8 column and pyridinecontaining mobile phases. Mar. Ecol. Prog. Ser., 195: 29-45. doi:10.3354/meps195029

Zingone, A., R. Siano, D. D’Alelio and D. Sarno. – 2006. Potentially toxic and harmful microalgae from coastal waters of the Campania region (Tyrrhenian Sea, Mediterranean Sea). Harmful Algae, 5: 321-337. doi:10.1016/j.hal.2005.09.002

Copyright (c) 2008 Consejo Superior de Investigaciones Científicas (CSIC)

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.

Contact us

Technical support