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formerly Central European Journal of Biology

Editor-in-Chief: Ratajczak, Mariusz

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Volume 10, Issue 1 (Nov 2014)


Isolation and preliminary characterization of cyanobacteria strains from freshwaters of Greece

Spyros Gkelis
  • Department of Botany, School of Biology, Aristotle University of Thessaloniki, GR-541 24 Thessaloniki, Macedonia, Greece
  • Email:
/ Pablo Fernández Tussy
  • Department of Botany, School of Biology, Aristotle University of Thessaloniki, GR-541 24 Thessaloniki, Macedonia, Greece
/ Nikos Zaoutsos
  • Department of Botany, School of Biology, Aristotle University of Thessaloniki, GR-541 24 Thessaloniki, Macedonia, Greece
Published Online: 2014-11-20 | DOI: https://doi.org/10.1515/biol-2015-0006


Cyanobacterial harmful algal blooms (or CyanoHABs) represent one of the most conspicuous waterborne microbial hazards. The characterization of the bloom communities remains problematic because the cyanobacterial taxonomy of certain genera has not yet been resolved. In this study, 29 planktic and benthic cyanobacterial strains were isolated from freshwaters located in Greece. The strains were assigned to the genera Chroococcus, Microcystis, Synechococcus, Jaaginema, Limnothrix, Pseudanabaena, Anabaena, and Calothrix and screened for the production of the cyanotoxins microcystins (MCs), cylindrospermopsins (CYNs), and saxitoxins (STXs) using molecular (PCR amplification of seven genes implicated in cyanotoxin biosynthesis) and immunological (ELISA) methods. This study presents, for the first time, a cyanobacteria culture collection from Greece, thus providing missing study material for the understanding of bloom formation and cyanotoxin production in the Mediterranean and for the polyphasic characterization of important components of the phytoplankton. The combined use of molecular and immunochemical methods allowed the identification of MC producing strains, but further data are needed for CYNand STX-producing cyanobacteria. The high percentage of MC-producing Microcystis strains in the urban Lakes Kastoria and Pamvotis, frequently used for agriculture irrigation, fishing and recreation, highlights the potential risk for human health.

Keywords: Microcystis; Anabaena; Limnothrix; Calothrix; cyanotoxins; molecular detection; lakes; ELISA


  • [1] Cook C.M., Vardaka E., Lanaras T., Toxic cyanobacteria in Greek freshwaters, 1987-2000: Occurrence, toxicity and impacts in the Mediterranean region., Acta Hyrdoch. Hydrob. 2004, 32, 107-124

  • [2] Paerl H.W., Hall N.S., Calandrino E.S., Controlling harmful cyanobacterial blooms in a world experiencing anthropogenic and climatic-induced change, Sci. Total Environ., 2011, 409, 1739-1745

  • [3] Paerl H.W., Otten G.T., Harmful Cyanobacterial Blooms: Causes, Consequences, and Controls, Microb. Ecol., 2013, 65, 995-1010

  • [4] Vardaka E., Moustaka-Gouni M., Cook C.M., Lanaras T., Cyanobacterial blooms and water quality in Greek waterbodies. J. Appl. Phycol., 2005, 17, 391-401 [Crossref]

  • [5] Gkelis S., Zaoutsos N., Cyanotoxin occurrence and potentially toxin producing cyanobacteria in freshwaters of Greece: a multi-disciplinary approach, Toxicon, 2014, 78, 1-9 [Crossref]

  • [6] Moustaka-Gouni M., Vardaka E., Michaloudi E., Kormas K.A., Tryfon E., Mihalatou H., Gkelis S., Lanaras T. , Plankton food web structure in a eutrophic polymictic lake with a history in toxic cyanobacterial blooms, Limnol. Oceanogr., 2006, 51, 715-727

  • [7] Moustaka-Gouni M., Kormas K.A., Vardaka E., Katsiapi M., Gkelis S., Raphidiopsis mediterranea Skuja represents non-heterocytous life-cycle stages of Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju in Lake Kastoria (Greece), its type locality: Evidence by morphological and phylogenetic analysis, Harmful Algae, 2009, 8, 864-872 [Crossref]

  • [8] Rajaniemi P., Hrouzek P., Kaštovská K., Willame R., Rantala A., Hoffmann L., Komarek J., Sivonen K., Phylogenetic and morphological evaluation of the genera Anabaena, Aphanizomenon, Trichormus and Nostoc (Nostocales, Cyanobacteria), Int. J. Syst. Evol. Micr., 2005, 55, 11-26 [Crossref]

  • [9] Willame R., Boutte C., Grubisic S., Wilmotte A., Komárek J., Hoffmann L., Morphological and molecular characterization of planktonic cyanobacteria from Belgium and Luxemburg., J. Phycol., 2006, 42, 1312-1332 [Crossref]

  • [10] Ballot A., Fastner J., Wiedner C., Paralytic shellfish poisoning toxin-producing cyanobacterium Aphanizomenon gracile in northeast Germany, Appl. Environ. Microbiol., 2010, 76, 1173-1180.

  • [11] Kling H.J., Laughinghouse H.D., Šmarda J., Komárek J., Acreman J., Bruun K., Watson S.B., Chen F., A new red colonial Pseudanabaena (Cyanoprokaryota, Oscillatoriales) from North American large lakes, Fottea, 2012, 12, 327-339 [Crossref]

  • [12] Anagnostidis K., Komárek J., Modern approach to the classification system of cyanophytes. 1- Introduction, Arch. Hydrobiol., 1985, 71, 291-302

  • [13] Anagnostidis K. & Komárek J., Modern approach to the classification system of cyanophytes. 3- Oscillatoriales, Arch. Hydrobiol., 1988, 80, 327-472

  • [14] Komárek J., Anagnostidis K., Modern approach to the classification system of cyanophytes. 2- Chroococales. Arch. Hydrobiol., 1986, 73, 157-226

  • [15] Komárek J., Anagnostidis K., Modern approach to the classification-system of cyanophytes 4 - Nostocales. Arch. Hydrobiol., 1989, 82: 247-345

  • [16] Castenholz R.W., Oxygenic photosynthetic bacteria. In: Boone D.R. & R. W. Castenholz (Eds), Bergey’s Manual of Systematic Bacteriology, vol. 1, p. 474-600, Springer-Verlag, New York, 2001

  • [17] Komárek J., Mareš J., An update to modern taxonomy (2011) of freshwater planktic heterocytous cyanobacteria, Hydrobiologia, 2012, 698, 327-351

  • [18] Gkelis S., Rajaniemi P., Vardaka E., Moustaka-Gouni M., Lanaras T., Sivonen K., Limnothrix redekei (Van Goor) Meffert (cyanobacteria) strains from Lake Kastoria form a separate phylogenetic group, Microb. Ecol., 2005, 49, 176-182

  • [19] Gkelis S., Harjunpää V., Lanaras T., Sivonen K., Diversity of hepatotoxic microcystins and bioactive anabaenopeptins in cyanobacterial blooms from Greek freshwaters, Environ. Toxicol., 2005, 20, 249-256 [Crossref]

  • [20] Rippka R., Isolation and purification of cyanobacteria, Method. Enzymol., 1988, 167, 3-27

  • [21] de Chazal N.M., Smaglinski S., Smith G.D., Methods involving light variation of cyanobacteria: Characterization of isolates from central Australia, Appl. Environ. Microb., 1992, 58, 3561-3566

  • [22] Dotsika E., Poutoukis D., Tzavidopoulosa I., Maniatis Y., Ignatiadou D., Raco B., A natron source at Pikrolimni Lake in Greece? Geochemical evidence, J. Geochem. Explor., 2009, 103, 133-143

  • [23] Komárek J., Anagnostidis K., Cyanoprokaryota. 1. Teil: Chroococcales. Süßwasserflora von Mitteleuropa Band 19/1, p. 270-323, Spektrum Akademischer Verlag, 1999

  • [24] Komárek J., Anagnostidis K., Cyanoprokaryota. 2. Teil: Oscillatoriales. Süßwasserflora von Mitteleuropa Band 19/2, p. 1-759, Spektrum Akademischer Verlag, 2005

  • [25] Atashpaz S., Khani S., Barzegari A., Barar J., Vahed S.Z., Azarbaijani, R., Omidi, Y., A robust universal method for extraction of genomic DNA from bacterial species. Microbiology, 2010, 79, 538-542 [Crossref]

  • [26] Vasconcelos V., Martins A., Vale M., Antunes A., Azevedo J., Welker M., Lopez O., Montejano G., First report on the occurrence of microcystins in planktonic cyanobacteria from Central Mexico, Toxicon, 2010, 56, 425-431 [Crossref]

  • [27] Tryfon E., Moustaka-Gouni M., Nikolaidis G., Planktic cyanophytes and their ecology in the shallow Lake Mikri Prespa, Greece, Nord. J. Bot., 1997, 17, 439-448 [Crossref]

  • [28] Mur L.R., Skulberg O.M., Utkilen H., Cyanobacteria in the environment, In: I Chorus and J Bartram (Eds) Toxic Cyanobacteria in Water. E & FN Spon, London, UK, pp 41-112, 1999

  • [29] Nonneman D., Zimba P., A PCR-based test to assess the potential for microcystin occurrence in channel catfish production ponds, J. Phycol., 2002, 38, 230-234 [Crossref]

  • [30] Tillett D., Dittmann E., Erhard M., Döhren H.V., Börner T., Neilan B.A., Structural organization of microcystin biosynthesis in Microcystis aeruginosa PCC7806: an integrated peptide polyketide synthetase system, Chem Biol, 2000. 7. 753-764 [Crossref]

  • [31] Ouahid Y., Perez-Silva G., Del Campo F.F., Identification of potentially toxic environmental Microcystis by individual and multiple PCR amplification of specific microcystin synthetase gene regions, Environ. Toxicol., 2005, 20, 235-242 [Crossref]

  • [32] Tillett D., Parker D.L., Neilan B.A., Detection of toxigenicity by a probe for the microcystin synthetase A gene (mcyA) of the cyanobacterial genus Microcystis: comparison of toxicities with 16S rRNA and phycocyanin operon (Phycocyanin Intergenic Spacer) phylogenies, Appl. Environ. Microbiol., 2001, 67, 2810-2818

  • [33] Jungblut A.D., Neilan B.A., Molecular identification and evolution of the cyclic peptide hepatotoxins, microcystin and nodularin, synthetase genes in three orders of cyanobacteria, Arch. Microbiol., 2006, 185, 107-114

  • [34] Sivonen K, Jones G., Cyanobacterial toxins. In: I Chorus and J Bartram (Eds) Toxic Cyanobacteria in Water. E & FN Spon, London, UK, pp 113-153, 1999

  • [35] Frazão B., Martins R., Vasconcelos V., Are Known Cyanotoxins Involved in the Toxicity of Picoplanktonic and Filamentous North Atlantic Marine Cyanobacteria? Mar. Drugs, 2010, 8, 1908-1919 [Crossref]

  • [36] Vareli K., Zarali E., Zacharioudakis G.S.A., Vagenas G., Varelis V., Pilidis G., Briasoulis E., Sainis I., Microcystin producing cyanobacterial communities in Amvrakikos Gulf (Mediterranean Sea, NW Greece) and toxin accumulation in mussels (Mytilus galloprovincialis), Harmful Algae, 2012, 15, 109-118 [Crossref]

  • [37] Mihali, T.K., Kellmann, R., Muenchhoff, J., Barrow K.D., Neilan B.A., Characterization of the gene cluster responsible for cylindrospermopsin biosynthesis, Appl. Environ. Microbiol., 2008, 74, 716-722 [Crossref]

  • [38] Wood S.A., Rasmussen J.P., Holland P.T., Campbell R., Crowe A.L.M., First report of the cyanotoxin anatoxin-a from Aphanizomenon issatschenkoi (Cyanobacteria), J. Phycol., 2007, 43, 356-365 [Crossref]

  • [39] Ballot, A., Ramm, J., Rundberget, T., Kaplan-Levy, R.N., Hadas, O., Sukenik, A., Wiedner C., Occurrence of non-cylindrospermopsin- producing Aphanizomenon ovalisporum and Anabaena bergii in Lake Kinneret (Israel), J. Plankton Res., 2011, 33, 1736-1746 [Crossref]

  • [40] Stücken, A., Jakobsen, K.S., The cylindrospermopsin gene cluster of Aphanizomenon sp. strain 10E6: organization and recombination, Microbiology+, 2010, 156, 2438–2451

  • [41] Mazmouz R., Chapuis-Hugon F., Mann S., Pichon V., Méjean A., Ploux O., Biosynthesis of cylindrospermopsin and 7-epicylindrospermopsin in Oscillatoria sp. strain PCC 6506: identification of the cyr gene cluster and toxin analysis, Appl. Environ. Microbiol., 2010, 67, 4943–4949

  • [42] Schembri M., Neilan B., Saint C., Identification of genes implicated in toxin production in the cyanobacterium Cylindrospermopsis raciborskii, Environ.Toxicol., 2001, 16, 413-421 [Crossref]

  • [43] Spoof L., Berg K.A., Rapala J., Laht, K., Lepisto L., Metcalf J.S., Codd G.A., Meriluoto J., First observation of cylindrospermopsin in Anabaena lapponica isolated from the Boreal Environment (Finland), Environ. Toxicol., 2006, 21, 552-560 [Crossref]

  • [44] Kellmann R., Mihali T.K., Jeon Y.J., Pickford R., Pomati F., Neilan B.A., Biosynthetic intermediate analysis and functional homology reveal a saxitoxin gene cluster in cyanobacteria, Appl. Environ. Microb., 2008, 74, 4044-4053 [Crossref]

  • [45] Dittmann E., Fewer D.P., Neilan B.A, Cyanobacterial toxins: biosynthetic routes and evolutionary routes, FEMS Microbiol. Rev., 2013, 37, 23-43 [Crossref]

  • [46] Hisbergues M., Christiansen G., Rouhiainen L., Sivonen K., Borner T., PCR-based identification of microcystin-producing genotypes of different cyanobacterial genera, Arch. Microbiol., 2003, 180, 402-410

  • [47] Fergusson K.M., Saint C.P, Multiplex PCR assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria, Environ. Toxicol., 2003, 18, 120-125 [Crossref]

About the article

Received: 2014-01-28

Accepted: 2014-08-21

Published Online: 2014-11-20

Citation Information: Open Life Sciences, ISSN (Online) 2391-5412, DOI: https://doi.org/10.1515/biol-2015-0006. Export Citation

©2015 Spyros Gkelis et al.. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. (CC BY-NC-ND 3.0)

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