Skip to main content
SpringerLink
Log in

Genetic variability of the invasive cyanobacteria Cylindrospermopsis raciborskii from Bir M’cherga reservoir (Tunisia)

  • Original Paper
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

Cylindrospermopsis raciborskii is an invasive freshwater cyanobacteria of tropical origin, also found in temperate regions. Due to its known ability to produce potent toxins, such as cylindrospermopsin and the paralytic shellfish poisoning, this species is of major concern from a water quality perspective. This study presents a genetic characterization of four C. raciborskii strains isolated from the Bir M’cherga Tunisian reservoir. The toxicity assessment was investigated via molecular biology tools, which suggested that all the isolated strains were not producing cylindrospermopsin, saxitoxin, or microcystin. This result was further confirmed by HPLC and MALDI-TOF analyses. However, we report for the first time in C. raciborskii the presence of mcyA and mcyE, two segments of the microcystin synthetase mcy cluster. All the strains were identified taxonomically based on the 16S rRNA sequences, and their phylogenetic relationships were assessed using the rpoC1 region. Tunisian strains formed a distinct clade separated from the other African strains.

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

Similar content being viewed by others

References

  • Aboal M, Puig M-A (2005) Intracellular and dissolved microcystin in reservoirs of the river Segura basin, Murcia, SE Spain. Toxicon 45:509–518

    Article  PubMed  CAS  Google Scholar 

  • Alster A, Kaplan-Levy RN, Sukenik A, Zohary T (2010) Morphology and phylogeny of a non-toxic invasive Cylindrospermopsis raciborskii from a Mediterranean Lake. Hydrobiologia 639:115–128

    Article  Google Scholar 

  • Bernard C, Harvey M, Briand JF, Bire R, Krys S, Fontaine JJ (2003) Toxicological comparison of diverse Cylindrospermopsis raciborskii strains, evidence of liver damage caused by a French C. raciborskii strain. Environ Toxicol 18:176–186

    Article  PubMed  CAS  Google Scholar 

  • Bouaïcha N, Nasri AB (2004) First report of cyanobacterium Cylindrospermopsis raciborskii from Algerian freshwaters. Environ Toxicol 19:541–543

    Article  PubMed  Google Scholar 

  • Bourke ATC, Hawes RB, Neilson A, Stallman ND (1983) An outbreak of the hepatoenteritis (the Palm Island mystery disease) possibly caused by algal intoxication. Toxicon 3:45–48

    Article  Google Scholar 

  • Briand JF, Jacquet S, Bernard C, Humbert JF (2003) Health hazards for terrestrial vertebrates from toxic cyanobacteria in surface water ecosystems. Vet Res 34:361–377

    Article  PubMed  CAS  Google Scholar 

  • Carmichael WW (1994) The toxins of cyanobacteria. Sci Am 270:78–86

    Article  PubMed  CAS  Google Scholar 

  • Chapman AD, Schelske CL (1997) Recent appearance of Cylindrospermopsis (Cyanobacteria) in five hypereutrophic Florida lakes. J Phycol 33:191–195

    Article  Google Scholar 

  • Chonudomkul D, Yongmanitchai W, Threeragool G, Kawachi M, Kasai F, Kaya K, Watanabe MM (2004) Morphology, genetic diversity, temperature tolerance and toxicity of Cylindrospermopsis raciborskii (Nostocales, Cyanobacteria) strains from Thailand and Japan. FEMS Microbiol Ecol 48:345–355

    Article  PubMed  CAS  Google Scholar 

  • Druart JC, Briand JF (2002) First record of Cylindrospermopsis raciborskii (Woloszyn ′ ska) Seenayya et Subba Raju (Cyanobacteria) in a lotic system in France. Ann Limnol Int J Lim 38:339–342

    Article  Google Scholar 

  • Dyble J, Paerl HW, Neilan BA (2002) Genetic characterization of Cylindrospermopsis raciborskii (cyanobacteria) isolates from diverse geographic origins based on nifH and pcBA-IGS nucleotide sequence analysis. Appl Environ Microbiol 68:2567–2571

    Article  PubMed  CAS  Google Scholar 

  • Fastner J, Erhard M, Von-Döhren H (2001) Oligopeptide diversity within a natural population of Microcystis spp. (Cyanobacteria) by typing single colonies by matrix assisted laser desorption ionization–time of flight mass spectrometry. Appl Environ Microbiol 67:5069–5076

    Article  PubMed  CAS  Google Scholar 

  • Fastner J, Heinze R, Humpage AR, Mischke U, Eaglesham GK, Chorus I (2003) Cylindrospermopsin occurrence in two German lakes and preliminary assessment of toxicity and toxin production of Cylindrospermopsis raciborskii (Cyanobacteria) isolates. Toxicon 42:313–321

    Article  PubMed  CAS  Google Scholar 

  • Fathalli A, Jenhani A, Saker M, Moreira C, Romdhane MS, Vasconcelos VM (2010) First observation of the toxic and invasive cyanobacterium species Cylindrospermopsis raciborskii in Tunisian freshwaters. Toxicity assessment and molecular characterization. Fresenius Environ Bull 19:1074–1083

    CAS  Google Scholar 

  • Fergusson KM, Saint CP (2003) Multiplex PCR Assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria. Environ Toxicol 18:120–125

    Article  PubMed  CAS  Google Scholar 

  • Garcia C (2001) Isolamento e toxicologia de estirpes de Microcystis aeruginosa da Lagoa de Mira. MSc Thesis, Faculty of Sciences, Porto University

  • Gugger M, Molica R, Le Berre B, Dufour P, Bernard C, Humbert JF (2005) Genetic diversity of Cylindrospermopsis strains (Cyanobacteria) isolated from four continents. Appl Environ Microbiol 71:1097–1100

    Article  PubMed  CAS  Google Scholar 

  • Haande S, Rohrlack T, Ballot A, Røberg K, Skulberg R, Beck M, Wiedner C (2008) Genetic characterisation of Cylindrospermopsis raciborskii (Nostocales, Cyanobacteria) isolates from Africa and Europe. Harmful Algae 7:692–701

    Article  CAS  Google Scholar 

  • Hamilton PB, Ley LM, Dean S, Pick FR (2005) The occurrence of the cyanobacterium Cylindrospermopsis raciborskii in Constance Lake, an exotic cyanoprokaryote new to Canada. Phycologia 44:17–25

    Article  Google Scholar 

  • Hawkins PR, Runnegar MTC, Jackson ARB, Falconer IR (1985) Severe hepatotoxicity caused by the tropical cyanobacterium (blue-green algae) Cylindrospermopsis raciborskii (Woloszynska) Seenaya and Subba Raju isolated from a domestic water supply reservoir. Appl Environ Microbiol 50:1292–1295

    PubMed  CAS  Google Scholar 

  • Hawkins PR, Chandrasena NR, Jones GJ, Humpage AR, Falconer IR (1997) Isolation and toxicity of Cylindrospermopsis raciborskii from an ornamental lake. Toxicon 35:341–346

    Article  PubMed  CAS  Google Scholar 

  • Hisbergues M, Christiansen G, Rouhiainen L, Sivonen K, Börner T (2003) PCR-based identification of microcystin-producing genotypes of different cyanobacterial genera. Arch Microbiol 180:402–410

    Article  PubMed  CAS  Google Scholar 

  • Hong Y, Steinman A, Biddanda B, Rediske R, Fahnenstiel G (2006) Occurrence of the Toxin-producing Cyanobacterium Cylindrospermopsis raciborskii in Mona and Muskegon Lakes, Michigan. J Great Lakes Res 32:645–652

    Article  Google Scholar 

  • Istvanovics V, Shafik HM, Presing M, Juhos S (2000) Growth and phosphate uptake kinetics of the cyanobacterium, Cylindrospermopsis raciborskii (Cyanophyceae) in through flow cultures. Freshwater Biol 43:257–275

    Article  Google Scholar 

  • Izaguirre G, Jungblut AD, Neilan BA (2007) Benthic cyanobacteria (Oscillatoriacea) that produce microcystin-LR, isolated from four reservoirs in Southern California. Water Res 41:492–498

    Article  PubMed  CAS  Google Scholar 

  • Janse van Vuuren S, Kriel GP (2008) Cylindrospermopsis raciborskii, a toxic invasive cyanobacterium in South African freshwaters. Afr J Aquat Sci 33:17–26

    Article  Google Scholar 

  • Jungblut A-D, Neilan BA (2006) Molecular identification and evolution of the cyclic peptide hepatotoxins, microcystin and nodularin, synthetase genes in three orders of cyanobacteria. Arch Microbiol 185:107–114

    Article  PubMed  CAS  Google Scholar 

  • Kellmann R, Mills T, Neilan BA (2006) Functional modeling and phylogenetic distribution of putative cylindrospermopsin biosynthesis enzymes. J Mol Evol 62:267–280

    Article  PubMed  CAS  Google Scholar 

  • Kellmann R, Mihali TK, Jeon YJ, Pickford R, Pomati F, Neilan BA (2008) Biosynthetic intermediate analysis and functional homology reveal a saxitoxin gene cluster in cyanobacteria. Appl Environ Microbiol 74:4044–4053

    Article  PubMed  CAS  Google Scholar 

  • Lagos N, Onodera H, Zagatto PA, Andrinolo D, Azevedo SMFO, Oshima Y (1999) The first evidence of paralytic shellfish toxins in the freshwater cyanobacterium Cylindrospermopsis raciborskii, isolated from Brazil. Toxicon 37:1359–1373

    Article  PubMed  CAS  Google Scholar 

  • Li R, Carmichael WW, Brittain S, Eaglesham GK, Shaw GR, Mahakhant A, Noparatnaraporn N, Yongmanitchai W, Kaya K, Watanabe MM (2001) Isolation and identification of the cyanotoxin cylindrospermopsin and deoxy-cylindrospermopsin from a Thailand strain of Cylindrospermopsis raciborskii (Cyanobacteria). Toxicon 39:973–980

    Article  PubMed  CAS  Google Scholar 

  • Manti G, Mattei D, Messineo V, Melchiorre S, Bogialli S, Sechi N, Casiddu P, Luglié L, Di Brizio M, Bruno M (2005) First report of Cylindrospermopsis raciborskii in Italy. Harmful Algal News 28:8–9

    Google Scholar 

  • Messineo V, Melchiorre S, Di Corcia A, Gallo P, Bruno M (2010) Seasonal succession of Cylindrospermopsis raciborskii and Aphanizomenon ovalisporum Blooms with Cylindrospermopsin occurrence in the Volcanic Lake Albano, Central Italy. Environ Toxicol 25:18–27

    PubMed  CAS  Google Scholar 

  • Mihali TK, Kellmann R, Muenchhoff J, Barrow KD, Neilan BA (2008) Characterization of the gene cluster responsible for cylindrospermopsin biosynthesis. Appl Environ Microbiol 74:716–722

    Article  PubMed  CAS  Google Scholar 

  • Mohamed ZA (2007) First report of toxic Cylindrospermopsis raciborskii and Raphidiopsis mediterranea (Cyanoprokaryota) in Egyptian fresh waters. FEMS Microbiol Ecol 59:749–761

    Article  PubMed  CAS  Google Scholar 

  • Moore D, O’Donohue M, Garnett C, Critchley C, Shaw G (2005) Factors affecting akinete differentiation in Cylindrospermopsis raciborskii (Nostocales, Cyanobacteria). Freshwater Biol 50:345–352

    Article  Google Scholar 

  • Moustaka-Gouni M, Vardaka E, Tryfon E (2007) Phytoplankton species succession in a shallow Mediterranean lake (L. Kastoria, Greece): steady-state dominance of Limnothrix redekei, Microcystis aeruginosa and Cylindrospermopsis raciborskii. Hydrobiologia 575:129–140

    Article  Google Scholar 

  • Neilan BA, Jacobs D, DelDot T, Blackall LL, Hawkins PR, Cox PT, Goodman AE (1997) rRNA sequences and evolutionary relationships among toxic and nontoxic cyanobacteria of the genus Microcystis. Int J Syst Bacteriol 47:693–697

    Article  PubMed  CAS  Google Scholar 

  • Neilan BA, Saker ML, Fastner J, Törökna A, Burns BP (2003) Phylogeography of the invasive cyanobacterium Cylindrospermopsis raciborskii. Mol Ecol 12:133–140

    Article  PubMed  CAS  Google Scholar 

  • Nonneman D, Zimba PV (2002) A PCR-based test to assess the potential for microcystin occurrence in channel catfish production ponds. J Phycol 38:230–233

    Article  Google Scholar 

  • O.C.D.E. (1982) Eutrophisation des eaux, méthodes de surveillance, d’évaluation et de lutte. O.C.D.E., Paris, 165p

  • Ohtani I, Moore RE, Runnegar MTC (1992) Cylindrospermopsin: a potent hepatotoxin from the blue-green alga Cylindrospermopsis raciborskii. J Am Chem Soc 114:7941–7942

    Article  CAS  Google Scholar 

  • Ouahid Y, Pérez-Silva GF, del Campo F (2005) Identification of potentially toxic environmental Microcystis by individual and multiple PCR amplification of specific microcystin synthetase gene regions. Environ Toxicol 20:235–242

    Article  PubMed  CAS  Google Scholar 

  • Padisák J (1997) Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju, an expanding highly adaptive blue–green algal species, worldwide distribution and review of its ecology. Arch Hydrobiol 107(Suppl):563–593

    Google Scholar 

  • Padisák J (1998) Sudden and gradual responses of phytoplankton to global climate change: case studies from two large, shallow lakes (Balaton, Hungary, and the Neusiedlersee, Austria/Hungary). In: George DG, Jones JG, Puncochar P, Reynolds CS, Sutcliffe DW (eds) Management of lakes and reservoirs during global change. Kluwer Academic Publishers, London, pp 111–125

    Google Scholar 

  • Preußel K, Stüken A, Wiedner C, Chorus I, Fastner J (2006) First report on cylindrospermopsin producing Aphanizomenon flos-aquae (Cyanobacteria) isolated from two German lakes. Toxicon 47:156–162

    Article  PubMed  Google Scholar 

  • Ressom R, Soong FS, Fitzgerald J, Turczynowicz L, Saadi OE, Roder D, Maynard T, Falconer IR (1994) Health effects of toxic cyanobacteria (Blue-Green Algae). National Health and Medical Research Council. Australian Government Publishing Service, Canberra

    Google Scholar 

  • Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111:1–61

    Google Scholar 

  • Romo S, Miracle MR (1994) Population dynamics and ecology of subdominant phytoplankton species in a shallow hypertrophic lake (Albufera of Valencia, Spain). Hydrobiologia 273:37–56

    Article  Google Scholar 

  • Rücker J, Wiedner C, Zippel P (1997) Factors controlling the dominance of Planktothrix agardhii and Limnothrix redekeii in eutrophic shallow lakes. Hydrobiologia 342:107–115

    Article  Google Scholar 

  • Saker ML (2000) Cyanobacterial blooms in tropical north Queensland water bodies. PhD Thesis, James Cook University, Townsville, Australia

  • Saker ML, Eaglesham GK (1999) The accumulation of cylindrospermopsin from the cyanobacterium Cylindrospermopsis raciborskii in tissues of the Redclaw crayfish Cherax quadricarinatus. Toxicon 37:1065–1077

    Article  PubMed  CAS  Google Scholar 

  • Saker ML, Neilan BA (2001) Varied diazotrophies, morphologies, and toxicities of genetically similar isolates of Cylindrospermopsis raciborskii (Nostocales, Cyanophyceae) from northern Australia. Appl Environ Microbiol 67:1839–1845

    Article  PubMed  CAS  Google Scholar 

  • Saker ML, Thomas AD, Norton JH (1999) Cattle mortality attributed to the toxic cyanobacterium Cylindrospermopsis raciborskii in an outback region of north Queensland. Environ Toxicol 14:179–183

    Article  CAS  Google Scholar 

  • Saker ML, Nogueira ICG, Vasconcelos VM, Neilan BA, Eaglesham GK, Pereira P (2003) First report and toxicological assessment of the cyanobacterium Cylindrospermopsis raciborskii from Portuguese freshwaters. Ecotoxicol Environ Saf 55:243–250

    Article  PubMed  CAS  Google Scholar 

  • Schembri MA, Neilan BA, Saint CP (2001) Identification of genes implicated in toxin production in the cyanobacterium Cylindrospermopsis raciborskii. Environ Toxicol 16:413–421

    Article  PubMed  CAS  Google Scholar 

  • Shalev-Alon G, Sukenik A, Livnah O, Schwarz R, Kaplan A (2002) A novel gene encoding amidinotransferase in the cylindrospermopsin producing cyanobacterium Aphanizomenon ovalisporum. FEMS Microbiol Lett 209:87–91

    Article  PubMed  CAS  Google Scholar 

  • Tamura D, Nei K (2007) Phylogenetic and molecular evolutionary analyses were conducted using MEGA version 4

  • Vardaka E, Moustaka-Gouni M, Cook CM, Lanaras T (2005) Cyanobacterial blooms and water quality in Greek waterbodies. J Appl Phys 17:391–401

    Google Scholar 

  • Varkonyi ZZ, Farkas O, Garab T, Gombos Z (2000) The tolerance of cyanobacterium Cylindrospermopsis raciborskii to low-temperature photo-inhibition affected by the induction of polyunsaturated fatty-acid synthesis. Biochem Soc T 28:892–894

    Article  CAS  Google Scholar 

  • Vasconcelos V (2006) Eutrophication, toxic cyanobacteria and cyanotoxins: when ecosystems cry for help. Limnetica 25:425–432

    Google Scholar 

  • Welker M, Bickel H, Fastner J (2002) HPLC-PDA detection of cylindrospermopsin opportunities and limits. Water Res 36:4659–4663

    Article  PubMed  CAS  Google Scholar 

  • Wilson KM, Schembri MA, Baker PD, Saint CP (2000) Molecular characterisation of the toxic cyanobacterium Cylindrospermopsis raciborskii and design of a species-specific PCR. Appl Environ Microbiol 66:332–338

    Article  PubMed  CAS  Google Scholar 

  • Wood SA, Stirling DJ (2003) First identification of the cylindrospermopsin producing cyanobacterium Cylindrospermopsis raciborskii in New Zealand. New Zealand J Mar Freshw Res 37:821–828

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by U.R Ecosystèmes et Ressources Aquatiques, Institut National Agronomique de Tunisie and L.E.G.E, Centro de Investigação Marinha e Ambiental- CIIMAR/CIMAR, Portugal. A fellowship was also accorded to Afef FATHALLI from the Embassy of Portugal in Tunisia, Service of Co-operation and Cultural Action. This study was partially funded by FCT through projects PTDC/AMB/67075/2006 and PTDC/AAC-AMB/104983/2008.

Author information

Authors and Affiliations

  1. Unité de Recherche Ecosystèmes et Ressources Aquatiques, Institut National Agronomique de Tunisie, 43, Avenue Charles-Nicolle, 1082, Tunis Mahrajène, Tunisie

    Afef Fathalli, Amel Ben Rejeb Jenhani & Mohamed Romdhane

  2. Centro Interdisciplinar de Investigação Marinha e Ambiental- CIIMAR/CIMAR, Rua dos Bragas, 289, 4050-123, Porto, Portugal

    Cristiana Moreira, Joana Azevedo, Agostinho Antunes & Vitor Vasconcelos

  3. AnagnosTec GmbH, Am Mühlenberg 11, 14476, Potsdam, Germany

    Martin Welker

  4. FG Umweltmikrobiologie, Technische Universität Berlin, Franklinstr. 29, 10587, Berlin, Germany

    Martin Welker

  5. Departamento de Biologia, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre, 4169-007, Porto, Portugal

    Vitor Vasconcelos

Authors
  1. Afef Fathalli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amel Ben Rejeb Jenhani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Cristiana Moreira
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Joana Azevedo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Martin Welker
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mohamed Romdhane
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Agostinho Antunes
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Vitor Vasconcelos
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Vitor Vasconcelos.

Additional information

Communicated by Erko Stackebrandt.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fathalli, A., Jenhani, A.B.R., Moreira, C. et al. Genetic variability of the invasive cyanobacteria Cylindrospermopsis raciborskii from Bir M’cherga reservoir (Tunisia). Arch Microbiol 193, 595–604 (2011). https://doi.org/10.1007/s00203-011-0704-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00203-011-0704-y

Keywords

Search

Navigation