Skip to main content
SpringerLink
Log in

Effects of nitrogen supply on Pseudo-nitzschia calliantha and Pseudo-nitzschia cf. seriata: field and laboratory experiments

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

The effects of inorganic and organic nitrogen supply on the growth and domoic acid (DA) production of Pseudo-nitzschia cf. seriata and Pseudo-nitzschia calliantha from Bizerte Lagoon (SW Mediterranean Sea) were studied during field and laboratory experiments. Nitrogen enrichments (40 µM NO3 −; 10 µM NH4 +; 20 µM CH4N2O) and a control, with no added N, were carried out in separate carboys with seawater collected from Bizerte Lagoon. In the field experiments, all N-enrichments resulted in significant increases in chlorophyll a concentration, and maintained exponential growth until the end of the experiment. The initial diatom community was dominated by a bloom of P. cf. seriata (9.3 × 105 cells l−1). After 6 days of incubation, the abundance of P. cf. seriata was greatest in the urea addition (1.52 × 106 cells l−1), compared to the ammonium treatment (0.47 × 106 cells l−1), nitrate treatment (0.70 × 106 cells l−1) and control (0.36 × 106 cells l−1). The specific growth rates, calculated from increases in chlorophyll a and cell abundance, were statistically different across all treatments, with the highest in the urea and nitrate additions. Similar results were obtained from the laboratory experiments. These were carried out with P. calliantha isolated from Bizerte Lagoon and grown in f/2 medium enriched with 40 µM nitrate, 10 µM ammonium and 20 µM urea. The exponential growth rate was significantly faster for the cells cultured with urea (1.50 d−1) compared to the nitrate (0.90 d−1) and ammonium (0.80 d−1) treatments and the control (0.40 d−1). Analysis of DA, performed at the beginning and the end of the both experiments in all treatments, revealed very low concentrations (below the limit of quantification, 0.02- 1.310−7 pg cell−1, respectively).The field and laboratory experiments demonstrate that P.cf. seriata and P. calliantha are able to grow efficiently on the three forms of N, but with a preference for urea.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Agawin NSR, Duarte CM, Agusti S (2000) Nutrient and temperature control of the contribution of picoplankton to phytoplankton biomass and production. Limnol Oceanogr 45:591–600

    Article  CAS  Google Scholar 

  • Almandoz GO, Ferrario ME, Ferreyra GA, Schloss IR, Esteves JL, Paparazzo FE (2006) The genus Pseudonitzschia (Bacillariophyceae) in continental shelf waters of Argentina (Southwestern Atlantic Ocean, 38–55°S). Harmful Algae 6:103–193

    Google Scholar 

  • Aminot A, Chausspied M (1983) Manuel des analyses chimiques en milieu marin Centre national pour l’exploitation des océans. Brest. ISBN:2. 902721.10.2

  • Bates SS, Davidovich NA (2002) Factors affecting the sexual reproduction of diatoms with emphasis on Pseudonitzschia spp LIFEHAB Life history of microalgal species causing harmful blooms. Workshop Report European Commission, pp 31–36

  • Bates SS, Worms J, Smith JC (1993) Effects of ammonium and nitrate on growth and domoic acid production by Nitzschia pungens in batch culture. Can J Fish Aquat Sci 50:1248–1254

    Article  CAS  Google Scholar 

  • Bates SS, Garrison DL, Horner RA (1998) Bloom dynamics and physiology of domoic acid- producing Pseudonitzschia species. In: Anderson DM, Cembella AD, Hallegraeff GM (eds) Physiological ecology of harmfulalgal blooms. Springer, Verlag, pp 267–292

  • Berg P, Roy HF, Janssen V, Meyer B, Jorgensen B, Huettel M, Beer D (2003) Oxygen uptake by aquatic sediments measured with a novel non-invasive eddy correlation technique. Mar Ecol Prog Ser 261:75–83

    Article  Google Scholar 

  • Bradley BA, Blumenthal DM, Wilcove DS, Ziska LH (2010) Predicting plant invasions in an era of global change. Trends Ecol Evol 25:310–318

    Article  Google Scholar 

  • Busse LB, Venrick EL, Antrobus R, Miller PE, Vigilant V, Silver MW, Mengelt C, Mydlarz L, Prezelin BB (2006) Domoic acid in phytoplankton and fish in San Diego CA USA. Harmful Algae 5:91–101

    Article  CAS  Google Scholar 

  • Calu G, Martin-Jezequel V, Lefau E, Sechet V, Lassus P, Weigel P, Amzil Z, (2009) The influence of nitrogen speciation on growth and toxicity of Pseudo-nitzschia multiseries and P pungens in batch and continuous cultures. In: Lassus P (ed) 7th international conference on molluscan shellfish safety Nantes France, p 7

  • Chikhaoui MA, Sakka Hlaili A, Hadj Mabrouk H (2008) Réponses saisonnières du phytoplancton aux rapports d’enrichissements N:Si: P dans la lagune de Bizerte (Sud-Ouest de la Méditerranée). C R Biol 331:389–408

    Article  CAS  Google Scholar 

  • Christensen T (1988) Alger i naturen og i laboratoriet Kobenhavns Universitet Institut for Sporeplanter. 137 pp

  • Cochlan WP, Herndon J, Kudela RM (2008) Inorganic and organic nitrogen uptake by the toxigenic diatom Pseudo-nitzschia australis (Bacillariophyceae). Harmful Algae 8:111–118

    Article  CAS  Google Scholar 

  • Collos Y, Bec Jauzein C, Abadie E, Laugier T, Lautier J, Pastoureaud A, Souchu P, Vaquer A (2014) Oligotrophication and emergence of picocyanobacteria and a toxic dinoflagellate in Thau lagoon southern France. J Sea Res 61:68–75

    Article  Google Scholar 

  • Couture JY, Levasseur M, Bonneau E, Desjardins C, Sauve G, Bates SS, Leger C, Gagnon J, Michaud S (2001) Spatial and temporal variation of domoic acid in molluscs and of Pseudo-nitzschia spp blooms in the St Lawrence from 1998 to 2000. Can Tech Rep Fish Aquat Sci 2375 vii 24 p

  • De la Broise D, Palenik B (2007) Immersed in situ microcosms: a tool for the assessment of pollution impact on phytoplankton. J Exp Mar Biol Ecol 341:274–281

    Article  Google Scholar 

  • De la Iglesia P, Giménez G, Diogène J (2008) Determination of dissolved domoic acid in seawater with reversed-phase extraction disks and rapid resolution liquid chromatography tandem mass spectrometry with head column trapping. J Chromatography A1215:116–124

    Article  Google Scholar 

  • Fritz L, Quilliam MA, Wright JLC, Beale A, Work TM (1992) An outbreak of domoic acid poisoning attributed to the pennate diatom Pseudo-nitzschia australis. J Phycol 28:439–442

    Article  Google Scholar 

  • Garcıa-Mendoza E, Rivas D, Olivos-Ortiz A, Almazan-Becerril A, Castaneda-Vega C, Pena-Manjarrez JL (2009) A toxic Pseudo-nitzschia bloom in Todos Santos Bay northwestern Baja California Mexico. Harmful Algae 8:493–503

    Article  Google Scholar 

  • Glibert PM, Garside C, Fuhrman JA, Roman MR (1991) Time-dependent coupling of inorganic and organic nitrogen uptake and regeneration in the plume of the Chesapeake Bay estuary and its regulation by large heterotrophs. Limnol Oceanogr 36:895–909

    Article  Google Scholar 

  • Glibert PM, Anderson DM, Gentien P, Granéli E, Sellner KG (2005) The global complex phenomena of harmful algal blooms. Oceanography 18:41–103

    Google Scholar 

  • Glibert PM, Seitzinger S, Heil CA, Burkolder JM, Parrow MW, Codispoti LA, Kelly V (2006) Escalating worldwide use of urea—a global change contributing to coastal eutrophication. Oceanography 18:198–208

    Article  Google Scholar 

  • Gobler CJ, Sanuado-Wilhelmy SA (2002) Effects of organic carbon organic nitrogen inorganic nutrients and iron additions on the growth of phytoplankton and bacteria during a brown tide bloom. Mar Ecol Prog Ser 209:19–34

    Article  Google Scholar 

  • Guillard RRL, Ryther JH (1962) Studies of marine planktonic diatoms, I, Cyclotella nana (Hustedt) and Detonula confervacea (Cleve) Gran Can. J Microb 8:229–239

    CAS  Google Scholar 

  • Hasle GR, Fryxell GA (1970) Diatoms: cleaning and mounting for light and electron microscopy Trans. Am Microsc Soc 89:469–474

    Article  Google Scholar 

  • Hillebrand H, Sommer U (1996) Nitrogenous nutrition of the potentially toxic diatom Pseudo-nitzschia pungens f, multiseries Hasle. J Plankton Res 18:295–301

    Article  Google Scholar 

  • Howard MDA, Cochlan WP, Ladizinsky N, Kudela RM (2007) Nitrogenous preference of toxigenic Pseudonitzschia australis (Bacillariophyceae) from field and laboratory experiments. Harmful Algae 6:206–217

    Article  CAS  Google Scholar 

  • Irwin AJZV, Finkel OM, Schofield E, Falkowski PG (2006) Scaling up from nutrient physiology to the size structure of phytoplankton communities. J Plankton Res 28:459–471

    Article  Google Scholar 

  • Kilham P, Heckey (1980) The evolutionary ecology of phytoplankton In: Morris I (Ed) The physiological ecology of phytoplankton. Blackwell Scientific Publications, Boston, pp 571–597

  • Klein C, Claquin P, Bouchart V, Le Roy B, Veron B (2010) Dynamics of Pseudo-nitzschia spp and domoic acid production in a macrotidal ecosystem of the Eastern English Channel (Normandy France). Harmful Algae 9:218–226

    Article  CAS  Google Scholar 

  • Kudela R, Cochlan W (2000) Nitrogen and carbon uptake kinetics and the influence of irradiance for a red tide bloom off southern California. Aquat Microb Ecol 21:31–47

    Article  Google Scholar 

  • Kudela R, Pitcher G, Probyn T, Figueiras F, Moita T, Trainer V (2005) Harmful algal blooms in coastal upwelling systems. Oceanography 18:184–197

    Article  Google Scholar 

  • Kudela R, Lane J, Cochlan W (2008) The potential role of anthropogenically derived nitrogen in the growth of harmful algae in California USA. Harmful algae 8:103–110

    Article  CAS  Google Scholar 

  • Landry MR, Hassett RP (1982) Estimating the grazing impact of marine microzooplankton. Mar Biol 67:283–288

    Article  Google Scholar 

  • Lelong A, Hegaret H, Soudant P, Bates S (2012) Pseudo-nitzschia (Bacillariophyceae) species domoic acid and amnesic shellfish poisoning: revisiting previous paradigms. Phycologia 51:168–216

    Article  CAS  Google Scholar 

  • Litchman E, Klausmeier CA, Schofield OM, Falkowski PG (2007) The role of functional traits and trade-offs in structuring phytoplankton communities scaling from cellular to ecosystem level. Ecol Lett 10:1170–1181

    Article  Google Scholar 

  • Loureiro S, Garcés E, Fernández-Tejedor M, Vaqué D, Camp J (2009) Pseudonitzschia spp (Bacillariophyceae) and dissolved organic matter (DOM) dynamics in the Ebro Delta (Alfacs Bay NW Mediterranean Sea). Estuar Coast Shelf Sci 83:539–549

    Article  CAS  Google Scholar 

  • Lund JWG, Kipling C, LeCren ED (1958) The inverted microscope method of estimation algal numbers and the statistical basis of estimating by counting. Hydrobiol 11:143–170

    Article  Google Scholar 

  • Lundholm N (2016) Bacillariophyta In IOC-UNESCO Taxonomic Reference List of Harmful Micro algae Edited by Lundholm N Available from: http://www.marinespecies.org/hab/aphia.php?p=taxlist&pid=576884&tRank=220&rComp=%3D&context_in=30&vOnly=1 [Accessed Nov 148 2016]

  • Maldonado M, Hughes MP, Rue EL, Wells ML (2002) The effect of Fe and Cu on growth and domoic acidproduction by Pseudo-nitzschia multiseries and Pseudonitzschia australis. Limnol Oceanogr 47:515–526

    Article  CAS  Google Scholar 

  • Martin JL, Haya K, Wildish DJ (1993) Distribution and domoic acid content of Nitzschia pseudodelicatissima in the Bay of Fundy In: Smayda TJ, Shimizu Y (eds) Toxic phytoplankton blooms in the sea. Elsevier, New York, pp 613–618

  • McDonald SM, Sarno S, Zingone A (2007) Identifying Pseudo-nitzschia species in natural samples using genusspecific PCR primers and clone libraries. Harm Algae 6:849–860

    Article  CAS  Google Scholar 

  • Murphy J, Riley JP (1962) A modified single solution method for the determination of phosphate in natural waters. Anal Chem Acta 27:31–36

    Article  CAS  Google Scholar 

  • Pan Y, Subba Rao DV, Mann KH, Brown RG, Pocklington R (1996) Effects of silicate limitation on production of domoic acid a neurotoxin by the diatom Pseudo-nitzschia multiseries I, Batch culture studies. Mar Ecol Prog Ser 131:225–233

    Article  CAS  Google Scholar 

  • Parsons ER, Takahashi M, Hargrave B (1984) Biological oceanographic, 3rd edn. Processes Pergamon Press, Oxford, p 330

  • Parsons ML, Dortch Q, Turner RE (2002) Sedimentological evidence of an increase in Pseudo-nitzschia (Bacillariophyceae) abundance in response to coastal eutrophication. Limnol Oceanogr 47:551–558

    Article  Google Scholar 

  • Price NM, Harrison PJ (1987) A comparison of methods for the measurement of dissolved urea concentration in seawater. Mar Biol 92:307–319

    Article  Google Scholar 

  • Quijano-Scheggia S, Garces E, Sampedro N, van Lenning K, Flo E, Andree K, Fortuno JM, Camp J (2008) Identification and characterisation of the dominant Pseudo nitzschia species (Bacillariophyceae) along the NE Spanish coast (CataloniaNW Mediterranean). Sci Mar 72:343–359

  • Quiroga I (2006) Pseudo-nitzschia bloom in the bay of Banyuls-sur-mer Northwestern Mediterranean sea. Diatom Res 21:91–104

    Article  Google Scholar 

  • Sahraoui I, Sakka Hlaili A, Hadj Mabrouk H, Claude Léger C, Bates SS (2009) Blooms of the Diatom genus Pseudo-nitzschia H, Peragallo in Bizerte Lagoon (Tunisia SW Mediterranean). Diatom Res 24:175–190

    Article  Google Scholar 

  • Sahraoui I, Bates SS, Bouchouicha D, Hadj Mabrouk H, Sakka Hlaili A (2011) Toxic and potentially toxic Pseudonitzschia populations in Bizerte Lagoon (Tunisia SW Mediterranean) during 2006-2007 and first report ofdomoic acid production by Pseudo-nitzschia brasiliana Lundholm Hasle Fryxell. Diatom Res 26:293–303

    Article  Google Scholar 

  • Sakka Hlaili A, Chikhaoui MA, Grami B, Hadj Mabrouk H (2006) Effet of N and P supply on phytoplankton in Bizerte lagoon (Western Mediterranean). J Exp Mar Biol Ecol 14:281–288

  • Scholin CA, Gulland F, Doucette GJ, Benson S, Busman M, Chavez FP, Cordaro J, DeLong R, De Vogelaere A, Harvey J, Haulena M, Lefebvre K, Lipscomb T, Loscutoff S, Lowenstine LJ, Marin RIII, Miller PE, McLellan WA, Moeller PDR, Powell CL, Rowles T, Silvagni P, Silver M, Spraker T, Trainer V, Van Dolah FM (2000) Mortality of sea lions along the central California coast linked to a toxic diatom bloom. Nature 403:80–84

    Article  CAS  Google Scholar 

  • Souchu P, Smith VH, Laugier T, Fiandrino A, Benau L, Orsoni V, Collos Y, Vaquer A, Bec B (2010) Patterns in nutrient limitation and chlorophyll a along an anthropogenic eutrophication gradient in French Mediterranean coastal lagoons. Can J Fish Aquat Sci 67:743–753

    Article  CAS  Google Scholar 

  • Spatharis S, Danielidis DB, Tsirtsis G (2007) Recurrent Pseudo-nitzschia calliantha (Bacillariophyceae) and Alexandrium insuetum (Dinophyceae) winter blooms induced by agricultural runoff. Harmful Algae 6:811–822

    Article  Google Scholar 

  • Strickland JDH, Parsons TR (1968) A practical handbook of seawater analysis. Ottawa Fisheries Research Board Canada

  • Syrett PJ (1981) Nitrogen metabolism of microalgae. Can Bull Fish Aquat Sci 210:182–210

    Google Scholar 

  • Teng ST, Lim HC, Lim PT, Dao VH, Bates SS, Leaw CP (2014) Pseudo-nitzschia kodamae sp nov (Bacillariophyceae) a toxigenic species from the Strait of Malacca Malaysia. Harmful Algae 34:17–28

    Article  Google Scholar 

  • Thessen AE, Bowers HA, Stoecker DK (2009) Intra- and interspecies differences in growth and toxicity of Pseudonitzschia while using different nitrogen sources. Harmful Algae 8:792–810

    Article  CAS  Google Scholar 

  • Trainer VL, Adams NG, Bill BD, Stehr CM, Wekell JC, Moeller P, Busman M, Woodruff D (2000) Domoic acid production near California coastal upwelling zones June 1998. Limnol Oceanogr 45:1818–1833

    Article  CAS  Google Scholar 

  • Trainer VL, Hickey BM, Horner RA (2002) Biological and physical dynamics of domoic acid production off the Washington coast. Limnol Oceanogr 47:1438–1446

    Article  CAS  Google Scholar 

  • Trainer VL, Hickey BM, Bates SS (2008) Toxic diatoms. In: Walsh PJ, Smith SL, Fleming LE, Solo-Gabriele Gerwick WH (eds) Oceans and human health risks and remedies from the seas. Elsevier Science Publishers, New York, pp 219–237

  • Trainer VL, Bates SS, Lundholm N, Thessen AE, Cochlan WP, Adams NG, Trick CG (2012) Pseudo-nitzschia physiological ecology phylogeny toxicity monitoring and impacts on ecosystem health. Harmful Algae 14:271–300

    Article  Google Scholar 

  • Utermöhl H (1931) Neue Wege in der quantitativen Erfassung des Planktons (mit besonderer Berücksichtigung des Ultraplanktons). Verh Int Ver Theor Angew Limnol 5:567–596

    Google Scholar 

  • Vaulot D, Frisoni GF (1986) Phytoplanktonic productivity and nutrients in five Mediterranean lagoons. Oceanol Acta 9:57–63

    CAS  Google Scholar 

  • Wells ML, Trick CG, Cochlan WP, Hughes MP, Trainer VL (2005) Domoic acid: the synergy of iron copper and the toxicity of diatoms. Limnol Oceanogr 50:1908–1917

    Article  CAS  Google Scholar 

  • Wood ED, Armstong FAJ, Richards FA (1967) Determination of nitrate in sea water by cadmium-copper reduction to nitrite. J Mar Biol Ass UK 47:23–31

Download references

Acknowledgments

The authors thank the two anonymous reviewers for their constructive and helpful comments. Thus we express our gratitude to S. Bates for help in improves the English.

Author information

Authors and Affiliations

  1. Laboratoire de Phytoplanctonologie, Faculté des Sciences de Bizerte, Université de Carthage, Bizerte, Tunisia

    Sondes Melliti Ben Garali, Inès Sahraoui & Asma Sakka Hlaili

  2. Institut Supérieur de Pêche et d’Aquaculture de Bizerte, B.P.15.7080, Errimel, Bizerte, Tunisia

    Sondes Melliti Ben Garali, Mohamed Chalghaf & Jamel Ksouri

  3. Institut de Recherche et Technologie Agroalimentaire (IRTA), Ctra. Poble Nou, Km 5.5, Sant Carles de la Rapita, 43540, Tarragona, Spain

    Pablo de la Iglesia & Jorge Diogène

Authors
  1. Sondes Melliti Ben Garali
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Inès Sahraoui
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pablo de la Iglesia
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mohamed Chalghaf
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jorge Diogène
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jamel Ksouri
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Asma Sakka Hlaili
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sondes Melliti Ben Garali.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Melliti Ben Garali, S., Sahraoui, I., de la Iglesia, P. et al. Effects of nitrogen supply on Pseudo-nitzschia calliantha and Pseudo-nitzschia cf. seriata: field and laboratory experiments. Ecotoxicology 25, 1211–1225 (2016). https://doi.org/10.1007/s10646-016-1675-1

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-016-1675-1

Keywords

Search

Navigation