High proliferation of Pseudo-nitzschia cf. arenysensis in the Adriatic Sea: ecological and morphological characterisation


Published: Dec 1, 2020
Keywords:
Adriatic Sea Pseudo-nitzschia bloom SEM ribosomal genes
JASNA ARAPOV
https://orcid.org/0000-0001-7568-1929
MIA BUŽANČIĆ
https://orcid.org/0000-0003-3848-9561
ANTONELLA PENNA
SILVIA CASABIANCA
SAMUELA CAPELLACCI
FRANCESCA ANDREONI
SANDA SKEJIĆ
ANA BAKRAČ
MAJA STRAKA
JELENA JELENA MANDIĆ
ŽIVANA NINČEVIĆ GLADANA
Abstract

The aim of the present study isto characterise the diatom Pseudo-nitzschia community during a bloom period together with environmental conditions. High proliferation of Pseudo-nitzschia spp. was observed in September 2017 at the shellfish breeding area in the Krka River estuary (Central eastern Adriatic Sea). The peak of abundance (1.8 x 106 cells L-1) was recorded at 7 m depth, and the increased abundance persisted for four weeks.Morphological analyses of field samples based on scanning electron microscopy (SEM) revealed that Pseudo-nitzschia cf. arenysensis was prevailing (94%) in the Pseudo-nitzschia assemblage. Several strains were successfully isolated from net samples in order to better define morphological features and phylogenetic characterisation. The isolated Pseudo-nitzschia strains corresponded morphologically to the P. cf. arenysensis from the field samples, based on our SEM observations. Phylogenetic analysis demonstrated that the Croatian strains grouped with P. arenysensis using the ITS and LSU rDNA sequences. Spearman rank correlation showed that salinity was an important environmental factor affecting the vertical distribution of Pseudo-nitzschia spp. in this highly variable area. Availability of increased concentration of orthophosphates and ammonium and low Si: TIN ratio may have promoted the bloom of P. cf. arenysensis in the estuary.

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Author Biographies
MIA BUŽANČIĆ, Institute of Oceanography and Fisheries, Croatia
Laboratory of Plankton and Shellfish Toxicity
ANTONELLA PENNA, University of Urbino, Department of Biomolecular Sciences, Italy
Department of Biomolecular Sciences
SILVIA CASABIANCA, University of Urbino, Department of Biomolecular Sciences, Italy
Department of Biomolecular Sciences
SAMUELA CAPELLACCI, University of Urbino, Department of Biomolecular Sciences, Italy
Department of Biomolecular Sciences
FRANCESCA ANDREONI, University of Urbino, Department of Biomolecular Sciences, Italy
Department of Biomolecular Sciences
SANDA SKEJIĆ, Institute of Oceanography and Fisheries, Croatia
Laboratory of Plankton and Shellfish Toxicity
ANA BAKRAČ, Institute of Oceanography and Fisheries, Croatia
Laboratory of Plankton and Shellfish Toxicity
MAJA STRAKA, Institute of Oceanography and Fisheries, Croatia
Laboratory of Plankton and Shellfish Toxicity
JELENA JELENA MANDIĆ, Institute of Oceanography and Fisheries, Croatia

Laboratory of Chemical Oceanography and Sedimentology of the Sea

 

 

ŽIVANA NINČEVIĆ GLADANA, Institute of Oceanography and Fisheries, Croatia
Laboratory of Plankton and Shellfish Toxicity
References
Adachi, M., Sako, Y., Ishida, Y., 1994. Restriction fragment length polymorphism of ribosomal DNA internal transcribed spacer and 5.8S regions in Japanese Alexandrium species (Dinophyceae). Journal of Phycology, 30, 857-863.
Ajani, P., Murray, S., Hallegraeff, G., Lundholm, N., Gillings, M. et al., 2013. The diatom genus Pseudo-nitzschia (Bacillariophyceae) in New South Wales, Australia: morphology, molecular phylogeny, and distribution. Journal of Phycology, 49, 765-785.
Ajani, P.A.,Verma, A., Lassudrie, M., Doblin, M.A., Murray, S.A., 2018. A new diatom species P. hallegraeffii sp. nov. belonging to the toxic genus Pseudo-nitzschia (Bacillariophyceae) from the East Australian Current. PLos ONE, 13, e0195622.
Arapov, J., Ujević, I., Marić Pfannkuchen, D., Godrijan, J., Bakrač, A. et al., 2016. Domoic acid in phytoplankton net samples and shellfish from the Krka River estuary in the Central Adriatic Sea. Mediterranean Marine Science, 17, 340-350.
Arapov, J., Skejić, S., Bužančić, M., Bakrač, A., Vidjak, O. et al., 2017. Taxonomical diversity of Pseudo-nitzschia from the Central Adriatic Sea. Phycological Research, 65, 280-290.
Bates, S.S., Hubbard, K.A., Lundholm, N., Montresor, M., Leaw, C.P., 2018. Pseudo-nitzschia, Nitzschia, and domoic acid: New research since 2011. Harmful Algae, 79, 3-43.
Bosak, S., Burić, Z., Djakovac, T., Viličić, D., 2009. Seasonal distribution of plankton diatoms in Lim Bay, northeastern Adriatic Sea. Acta Botanica Croatica, 68, 351-365.
Burić, Z., Viličić, D., Caput Mihelić, K., Carić, M., Kralj, K. et al., 2008. Pseudo-nitzschia blooms in the Zrmanja River estuary. Diatom Research, 23, 51-63.
Bužančić, M., Ninčević Gladan, Ž., Marasović, I., Kušpilić, G., Grbec, B. et al., 2012. Population structure and abundance of phytoplankton in three bays on the eastern Adriatic coast: Šibenik Bay, Kaštela Bay and Mali Ston Bay. Acta Adriatica, 53, 413-435.
Caroppo, C., Congestri, R., Bracchini, L., Albertano, P., 2005. On the presence of Pseudo-nitzschia calliantha Lundholm, Moestrup et Hasle and Pseudo-nitzschia delicatissima (Clave) Heiden in the Southern Adriatic Sea (Mediterranean Sea, Italy). Journal of Plankton Research, 27, 763-774.
Cetinić, I., Viličić, D., Burić, Z., Olujić, G., 2006. Phytoplankton seasonality in a highly stratified karstic estuary (Krka, Adriatic Sea). Hydrobiologia, 555, 31-40.
Ciminiello, P., Dell’Aversano, C., Fattorusso, E., Forino, M., Magno, G. S. et al., 2005. Hydrophilic interaction liquid chromatography/mass spectrometry for determination of domoic acid in Adriatic shellfish. Rapid Communications in Mass Spectrometry, 19, 2030-2038.
Clarke, K.R., Warwick, R.M., 2001. Changes in marine communities: an approach to statistical analysis and interpretation, 2nd edition. PRIMER-E: Plymouth. DHMZ, 2019. Croatian Meteorological and Hydrological Services. http://hidro.dhz.hr/ (Accessed 12 March 2019)
EU, 2004. Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for food of animal origin. Official Journal of the European Union L139, 22-82.
Grasshoff, K., Kremling, K., Ehrhardt, M., 1999. Methods of Seawater Analysis, third ed. Wiley-VCH, Weinheim, 419 pp.
Grbin, D., Pfannkuchen, M, Babić, I., Mejdandžić, M., Mihanović, H. et al., 2017. Multigene phylogeny and morphology of newly isolated strain of Pseudo-nitzschia mannii Amato & Montresor (Adriatic Sea). Diatom Research, 32, 127-131.
Hansen, L.R., Soylu, S., Kotaki, Y., Moestrup, Ø., Lundholm, N., 2011. Toxin production and temperature-induced morphological variation of the diatom Pseudo-nitzschia seriata from the Arctic. Harmful Algae, 10, 689-696.
Hasle, G.R., Fryxell, G.A., 1970. Diatoms: Cleaning and mounting for light and electron microscopy. Transactions of the American Microscopical Society, 89, 469-474.
Huang C.X., Dong, H.C., Lundholm, N., Teng, S.T., Zheng, G.C. et al., 2019. Species composition and toxicity of the genus Pseudo-nitzschia in Taiwan Strait, including P. chiniana sp. nov. and P. qiana sp. nov. Harmful Algae, 84, 195- 209.
Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung, M. et al., 2012. Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics, 28, 1647-1649.
Klein, C., Claquin, P., Bouchart, V., Le Roy, B., Véron, 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.
Kumar, S., Stecher, G., Tamura, K., 2016. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Big- ger Datasets. Molecular Biology and Evolution, 33, 1870-1874.
La Barre, S., Bates, S.S., Quilliam, M.A., 2014. Domoic Acid. p. 189-215. In: Outstanding Marine Molecules: Chemistry, Biology, Analyses.
La Barre, S., Kornprobost, J.-M. (Eds.). Wiley-VCH Verlag GmbH & Co. KGaA. Lelong, A., Hégaret, H., Soudant, P., Bates S.S., 2012. Pseudo- nitzschia (Bacillariophyceae) species, domoic acid and amnesic shellfish poisoning: revisiting previous paradigms. Phycologia, 51, 168-216.
Lim, H.C., Teng, S.T., Leaw, C.P., Lim, P.T., 2013. Three novel species in the Pseudo-nitzschia pseudodelicatissima complex: P. batesiana sp. nov., P. lundholmiae sp. nov., and P. fukuyoi sp. nov. (Bacillariophyceae) from the strait of Malacca, Malaysia. Journal of Phycology, 49, 902-916.
Loureiro, S., Jauzein, C., Garcés, E., Collos, Y., Camp, J. et al., 2009. The significance of organic nutrients in the nutrition of Pseudo-nitzschia delicatissima (Bacillaeriophyceae). Journal of Plankton Research, 31, 399-410.
Lundholm, N. (Ed), 2019. Bacillariophyceae, in IOC-UNESCO Taxonomic Reference List of Harmful Micro Algae. Available online at http://www.marinespecies.org/hab (Accessed 24 September 2019).
Lundholm, N., Moestrup, Ø., Kotaki, Y., Hoef-Emden, K., Scholin, C. et al., 2006. Inter- and intraspecific variation of the Pseudo-nitzschia delicatissima complex (Bacillariophyceae) illustrated by rRNA probes, morphological data and phylogenetic analyses. Journal of Phycology, 42, 464-481.
Ljubešić, Z., Bosak, S., Viličić, D., Kralj Borojević, K., Marić, D. et al., 2011. Ecology and taxonomy of potentially toxic Pseudo-nitzschia species in Lim Bay (north-eastern Adriatic Sea). Harmful Algae, 10, 713-722.
Macintyre, H.L., Stutes, A.L., Smith, W.L., Dorsey, C.P., Abraham, A. et al., 2011. Environmental correlates of community composition and toxicity during a bloom of Pseudo- nitzschia spp. in the Northern Gulf of Mexico. Journal of Plankton Research, 33, 273-295.
Marchetti, A., Harrison, P.J., 2007. Coupled changes in the cell morphology and the elemental (C, N, and Si) composition of the pennate diatom Pseudo-nitzschia due to iron deficiency. Limnology and Oceanography, 52 (5), 2270-2284.
Marchetti, A., Lundholm, N., Kotaki, Y., Hubbard, K., Harrison, P. J. et al., 2008. Identification and assessment of domoic acid production in oceanic Pseudo-nitzschia (Bacillariophyceae) from iron-limited waters in the northeast subarctic Pacific. Journal of Phycology, 44, 650-61.
Marić Pfannkuchen, D. 2013. Potentially toxic diatom genus Pseudo-nitzschia in the northern Adriatic Sea: ecological, taxonomic and molecular features. Ph.D. Thesis, University of Zagreb, Zagreb, Croatia, 195 pp.
Marić, D., Ljubešić, Z., Godrijan, J., Viličić, D., Ujević, I. et al., 2011. Blooms of the potentially toxic diatom Pseudo-nitzschia calliantha Lundholm, Moestrup & Hasle in coastal waters of the northern Adriatic Sea (Croatia). Estuarine, Coastal and Shelf Science, 92, 323-331.
Marić, D., Kraus, R., Godrijan, J., Supić, N., Djakovec, T. et al., 2012. Phytoplankton response to climatic and anthropogenic influences in the north-eastern Adriatic during the four last decades. Estuarine, Coastal and Shelf Science, 115, 98-112.
Montagnes, D.J.S., Franklin, D.J., 2001. Effects of temperature on diatom volume, growth rate, and carbon and nitrogen content: Reconsidering some paradigms. Limnology and Oceanography, 46 (8), 2008-2018.
Moschandreou, K.K., Nikolaidis, G., 2010. The genus Pseudo- nitzschia (Bacillariophyceae) in Greek coastal waters. Botanica Marina, 53, 159-172.
Moschandreou, K.K., Baxevanis, A.D., Katikou, P., Papaefthimiou, D., Nikolaidis, G. et al., 2012. Inter- and intra-specific diversity of Pseudo-nitzschia (Bacillariophyceae) in the northeastern Mediterranean. European Journal of Phycology, 47, 321-339.
Ninčević-Gladan, Ž., Bužančić, M., Kušpilić, G., Grbec, B., Matijević, S. et al., 2015. The response of phytoplankton community to anthropogenic pressure gradient in the coastal waters of the eastern Adriatic Sea. Ecological Indicators, 56, 106-115.
Ninčević Gladan, Ž., Matić, F., Arapov, J., Skejić, S., Bužančić, M. et al., 2020. The relationship between toxic phytoplankton species occurrence and environmental and meteorological factors along the Eastern Adriatic coast. Harmful Algae, 92.
Orive, E., Pérez-Aicuna, L., David, H., García-Etxebarria, K., Laza-Martínez, A. et al., 2013. The genus Pseudo-nitzschia (Bacillariophyceae) in a temperate estuary with description of two new species: Pseudo-nitzschia plurisecta sp. nov. and Pseudo-nitzschia abrensis sp. nov. Journal of Phycology, 49, 1192-1206.
Orlova, T.Y., Stonik, I.V., Aizdaicher, N.A., Bates, S.S., Léger, C. et al., 2008. Toxicity, morphology and distribution of Pseudo-nitzschia calliantha, P. multistriata and P. multiseries (Bacillariophyta) from the northwestern Sea of Japan. Botanica Marina, 51, 297-306.
Orsini, L., Sarno, D, Procaccini, G., Poletti, R., Dahlmann, J. et al., 2002. Toxic Pseudo-nitzschia multistriata (Bacillariophyceae) from the Gulf of Naples: morphology, toxin analyses and phylogenetic relationships with other Pseudo- nitzschia species. European Journal of Phycology, 37, 247-257.
Penna, A., Casabianca, S., Perini, F., Bastianini, M., Riccardi, E. et al., 2013. Toxic Pseudo-nitzschia spp. in the northwestern Adriatic Sea: characterization of species composition by genetic and molecular quantitative analyses. Journal of Plankton Research, 35, 352-366.
Pistocchi, R., Guerrini, F., Pezzolesi, L., Riccardi, M., Vanucci, S. et al., 2012. Toxin levels and profiles in microalgae from the North-western Adriatic Sea- 15 years of studies on cultured species. Marine Drugs, 10, 140-162.
Pugliese, L., Casabianca, S., Perini, F., Andreoni, F., Penna, A., 2017. A high resolution melting method for the molecular identification of the potentially toxic diatom Pseudo-nitzschia spp. in the Mediterranean Sea. Scientific Reports, 7, 4259.
Quijano-Scheggia, S., Garcés, E., Flo, E., Fernandez-Tejedor, M., Diogene, J. et al., 2008a. Bloom dynamics of the genus Pseudo-nitzschia (Bacillariophyceae) in two coastal bays (NW Mediterranean Sea). Scientia Marina, 72, 577-590.
Quijano-Scheggia, S., Garcés, E., Sampedro, N., Van Lenning, K., Flo, E. et al., 2008b. Identification and characterisation of the dominant Pseudo-nitzschia species (Bacillariophyceae) along the NE Spanish coast (Catalonia, NW Mediterranean). Scientia Marina, 72, 343-359.
Quijano-Scheggia, S., Garcés, E., Lundholm, N., Moestrup, Ø, Andree, K. et al., 2009. Morphology, physiology, molecular phylogeny and sexual compatibility of the cryptic Pseudo-nitzschia delicatissima complex (Bacillariophyta), including the description of P. arenysensis sp. nov. Phycologia, 48, 492-509.
Quijano-Scheggia, S., Garcés, E., Andree, K.B., Iglesia, P., Diogène, J. et al., 2010. Pseudo-nitzschia species on the Catalan coast: charcterization and contribution to the current knowledge of the distribution of this genus in the Mediterranean Sea. Scientia Marina, 74, 395-410.
Quilliam, M. A., Xie, M., Hardstaff, W. R., 1995. A rapid extraction and cleanup procedure for the liquid chromatographic determination of domoic acid in unsalted seafood. Journal of AOAC International, 78, 543-554.
Radan, R.L., Cochlan, W.P., 2018. Differential toxin response of Pseudo-nitzschia multiseries as a function of nitrogen speciation in batch and continuous cultures, and during a natural assemblage experiment. Harmful Algae, 73, 12-29.
Rivera-Vilarelle, M., Quijano-Scheggia, S., Olivos-Ortiz, A., Gaviño-Rodríguez, J.H., Castro-Pchoa, F. et al., 2013. The genus Pseudo-nitzschia (Bacillariophyceae) in Manzanillo and Santiago Bays, Colima, Mexico. Botanica Marina, 56, 357-373.
Ronquist, F., Huelsenbeck, J.P., 2003. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics, 19, 1572-1574.
Sahraoui, I., Hlaili, A.S., Mabrouk, H.H., Léger, C., Bates, S.S., 2009. Blooms of the diatom genus Pseudo-nitzschia H. Pergallo in Bizerte lagoon (Tunisia, SW Mediterranean). Diatom Research, 24, 175-190.
Sahraoui, I., Bates, S. S., Bouchouicha, D., Mabrouk, H. H., Hlaili, A.S., 2011. Toxicity of Pseudo-nitzschia population from Bizerte Lagoon, Tunisia, southwest Mediterranean, and first report of domoic acid production by P. brasiliana. Diatom Research, 26, 293-303.
Sahraoui, I., Grami, B., Bates, S.S., Bouchouicha, D., Chikhaoui, M.A. et al., 2012. Response of potentially toxic Pseudo-nitzschia (Bacillariophyceae) population and domoic acid to environmental conditions in a eutrophied, SW Mediterranean coastal lagoon (Tunisia). Estuarine, Coastal and Shelf Science, 102-103, 95-104.
Schlitzer, R., Ocean Data View, https://odv.awi.de, 2018. Scholin, C.A., Herzog, M., Sogin, M., Anderson, D.M., 1994. Identification of group and strain-specific using genetic markers from globally distributed Alexandrium (Dinophyceae). II.
Sequence analysis of fragments of the LSU rRNA gene. Journal of Phycology, 999-1011.
Skejić, S., Bojanić, N, Matijević, S., Vidjak, O., Grbec, B. et al., 2014. Analysis of the phytoplankton community in the vicinity of domestic sewage outflow during stratified conditions. Mediterranean Marine Science, 15, 574-586.
Stonik, I.V., Orlova, T.Y., Lundholm, N., 2011. Diversity of Pseudo-nitzschia H. Peragallo from the western North Pacific. Diatom Research, 26, 121-134.
Stonik, I.V., Isaeva, M.P., Aizdaicher, N.A., Balakirev, E.S., Ayala, F.J., 2018. Morphological and genetic identification of Pseudo-nitzschia H. Peragallo, 1900 (Bacillariophyta) from the Sea of Japan. Russian Journal of Marine Biology, 44, 192-201.
Su, Y., Lundholm, N., Ellegaard, M., 2018. Effects of abiotic factors on the nanostructure of diatom frustules-ranges and variability. Applied Microbiology and Biotechnology, 102, 5889-5899.
Svensen, C., Viličić, D., Wassmann, P., Arashkevich, E., Ratkova, T., 2007. Plankton distribution and vertical flux of biogenic matter during high summer stratification in the Krka river estuary (Eastern Adriatic). Estuarine, Coastal and Shelf Science, 71, 381-390.
Teng, S.T., Leaw, C.P., Lim, H.C., Lim, P.T., 2013. The genus Pseudo-nitzschia (Bacillariophyceae) in Malaysia, including new records and a key to species inferred from morphology- based phylogeny. Botanica Marina, 56, 375-398.
Teng, S.T., Tan, S.N., Lim, H.C., Dao, V.H., Bates, S.S. et al., 2016. High diversity of Pseudo-nitzschia along the northern coast of Sarawak (Malaysian Borneo), with descriptions of P. bipertita sp. nov. and P. limii sp. nov. (Bacillariophyceae). Journal of Phycology, 49, 973-989.
Thessen, A.E., Dortch, Q., Parsons, M.L., Morrison, W., 2005. Effect of salinity on Pseudo-nitzschia species (Bacillariophyceae) growth and distribution. Journal of Phycology, 41, 21-29.
Thorel, M., Claquin, P., Schapira, M., Le Gendre, R., Riou, P. et al., 2017. Nutrient ratios influence variability in Pseudo- nitzschia species diversity and particulate domoic acid production in the Bay of Seine (France). Harmful Algae, 68, 192-205.
Trainer, V.L., Bates, S.S., Lundholm, N., Thessen, A.E., Cochlan, W.P. et al., 2012. Pseudo-nitzschia physiological ecology, phylogeny, toxicity, monitoring and impacts on ecosystem health. Harmful Algae, 14, 271-300.
Turk Dermastia, T., Cerino, F., Stanković, D., Francé, J., Ramšak, A. et al. 2020. Ecological time series and integrative taxonomy unveil seasonality and diversity of the toxic diatom Pseudo-nitzschia H. Pergallo in the northern Adiratic Sea. Harmful Algae, 93.
Ujević, I., Ninčević Gladan, Ž., Roje, R., Skejić, S., Arapov, J. et al., 2010. Domoic acid – A New Toxin in the Croatian Adriatic Shellfish Toxin Profile. Molecules, 15, 6835-6849.
Utermöhl, H., 1958. Zur Vervollkommung der quantitative Phytoplankton Methodik. Internationale Vereinigung für Theoretische und angewandte Limnologie, 9, 1-38.
Viličić, D., Legović, T., Žutić, V., 1989. Vertical distribution of phytoplankton in a stratified estuary. Aquatic Sceinces, 51 (1), 31-46.
Viličić, D., Bosak, S., Burić. Z., Caput-Mihalić, K., 2007. Phytoplankton seasonality and composition along the coastal NE Adriatic Sea during the extremely low Po River discharge in 2006. Acta Botanica Croatica, 66, 101-115.
Viličić, D., Djakovac, T., Burić, Z., Bosak, S. 2009. Composition and annual cycle of phytoplankton assemblages in the northeastern Adriatic Sea. Botanica Marina, 52, 291-305.
Wright, J.L.C., Boyd, R. K., De Freitas, A.S.W., Falk, M., Foxall, R.A. et al., 1989. Identification of domoic acid, a neuroexcitatory amino acid, in toxic mussels from eastern Prince Edward Island. Canadian Journal of Chemistry, 67, 481-490.
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