Cyanobacterial Toxins in Freshwater Ecosystems
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Diversity, impact and fate of cyanobacterial toxins in freshwater ecosystems Dissertation submitted for the degree of Doctor of Natural Sciences (Dr. rer. nat.) Presented by SHIVA SHAMS At the Faculty of Sciences Department of Biology Konstanz, 2015 2 ____________________________________________________________________ “Nothing great in the world has been accomplished without passion.” – Georg Wilhelm Friedrich Hegel 3 I. Publications and Honours A. Publications Cerasino, L., Shams, S., Boscaini A., Salmaso, N., 2015. Inter-annual variability of the microcystins pool in the oligo-mesotrophic Lake Garda (Italy). In Preparation. Shams, S., Capelli, C., Cerasino, L., Ballot, A., Dietrich, D.R., Sivonen, K., Salmaso, N., 2015. Anatoxin-a producing Tychonema (cyanobacteria) in European water bodies. Water Research 69, 68-79. Shams, S., Cerasino, L., Salmaso, N., Dietrich, D.R., 2014. Experimental models of microcystin accumulation in Daphnia magna grazing on Planktothrix rubescens: Implications for water management. Aquatic Toxicology 148, 9-15. Salmaso, N., Copetti, D., Cerasino, L., Shams, S., Capelli, C., Boscaini, A., Valsecchi, L., Pozzoni, F., Guzzell, L., 2014.Variability of microcystin cell quota in metapopulations of Planktothrix rubescens: causes and implications for water management. Toxicon 90, 82-96. Jiang, L., Eriksson, J., Lage, S., Jonasson, S., Shams, S., Mehine, M., Ilag, L. Rasmussen, U., 2014. Diatoms: A Novel Source for the Neurotoxin BMAA in Aquatic Environments. PLoS One 9(1), e84578. Salmaso, N., Boscaini, A., Shams, S., Cerasino, L., 2013. Strict coupling between the development of Planktothrix rubescens and microcystin content in two nearby lakes south of the Alps (lakes Garda and Ledro). Annales de Limnologie - International Journal of Limnology 49 (4), 309- 318. 4 B. Conference talks July 1-5, 2013, Münster, Germany Shams, S., Cerasino, L., Salmaso, N., Dietrich, D.R. Experimental models of microcystin accumulation in Daphnia magna grazing on Planktothrix rubescens: potential for microcystin transfer through the food web. SEFS 8 (8th Symposium for European Freshwater Science). October 29 - November 2, 2012, Changwon, South Korea Shams, S., Cerasino, L., Salmaso, N., Dietrich, D.R. Diversity and Seasonality Of cyanotoxins In Lake Garda (Italy): Potential for Hepatotoxic Microcystins Transfer through the Food Web. 15th International Conference on Harmful Algae, Changwon, Republic of Korea. November 28-30, 2011, Vienna, Austria Shams, S., Cerasino, L., Salmaso, N. Diversity and seasonality of cyanotoxins in Lake Garda. 4th EULAKE meeting and science day, University of Natural Resources and Life Sciences. August 21-28, 2011, Trento, Italy Shams, S, Milan, M., Ranjan, J., Tolotti, M., Cerasino, L., Boscaini, A., Salmaso, N. Ecological changes of Central European Lakes within the EU Project EULAKES: nuisance cyanobacteria and cyanotoxins, and their impact on aquatic ecosystems and water quality at different temporal scales. 16th Workshop of the International Association of Phytoplankton Taxonomy and Ecology (IAP) held in San Michele all'Adige, Trento, Italy. C. Posters October 8-12, 2012, Konstanz, Germany Cerasino, L., Shams, S., Salmaso, N., Dietrich, D.R. Toxic potential of cyanobacteria in oligo- mesotrophic lakes: the case of Lake Garda (Italy). 3rd European Large Lakes Symposium University of Konstanz. June 26-July 1st, 2011, Girona, Spain Shams, S., Cerasino, L., Ranjan, J., Salmaso, N. Cyanotoxins and their impact on water quality in Central Europe lakes. The 7th Symposium of the European Freshwater Science, Girona, Spain. 5 D. Other conference contributions February 22-27, 2015, Granada, Spain Capelli, C., Shams, S., Cerasino, L., Cavalieri, D., Salmaso, N. Toxic cyanobacteria in the deep lakes south of the alps: A molecular assessment on cyanotoxin producing genotypes. Aquatic Sciences Meeting (ASLO). February 22-27, 2015, Granada, Spain Cerasino, L., Shams S., Boscaini A., Salmaso, N. Evolution of the toxin diversity in the oligo- mesotrophic environment of Lake Garda (Italy). Aquatic Sciences Meeting (ASLO). February 22-27, 2015, Granada, Spain Salmaso, N., Capelli, C., Shams, C., Boscaini, A., Tolotti, M., Cerasino, L. Invasion of Dolichospermum lemmermannii (cyanobacteria) to the deep lakes south of the alps: an unusual colonization from north to south? Aquatic Sciences Meeting (ASLO). September 1-5, 2014, Perugia, Italy Capelli, C., Shams, S., Cerasino, L., Cavalieri, D., Salmaso, N. Toxic cyanobacteria in lake Garda: a molecular assessment on cyanotoxin producing genotypes. 15th World Lake Conference (WLC15). September 1-5, 2014, Perugia, Italy Cerasino, L., Shams, S., Boscaini, A., Salmaso, N. Toxin dynamics inside the resident cyanobacterial community of Lake Garda (Italy). 15th World Lake Conference (WLC15). September 1-5, 2014, Perugia, Italy Salmaso, N., Tolotti, M., Milan, M., Shams, S., Capelli, C., Boscaini, A., Pareeth, S., Neteler, M., Cerasino, L. Impact of anthropogenic pressures and long-term climate changes on the trophic state of Lake Garda (Northern Italy): a multidisciplinary assessment. 15th World Lake Conference (WLC15). September 15-17, 2014, Ferrara, Italy Salmaso, N., Shams, S., Capelli, C., Boscaini, A., and Cerasino, L. From species to strains: production of toxins in populations of cyanobacteria, and implications for water management. Dipartimento di Scienze della Vita e Biotecnologie (SVeB), Università degli Studi di Ferrara, Italy. May 30, 2013, Gardone Riviera, Brescia, Italy Salmaso, N., Shams, S., Cerasino, L. Development of cyanobacteria and production of cyanotoxins in Lake Garda: statistical models and predictive power. Workshop of Quality and sustainable use of water resources in Lake Garda and in other large water bodies in Europe: 6 experiences within the project EULAKES. May 30, 2013, Gardone Riviera, Brescia, Italy Cerasino, L., Shams, S., Salmaso, N. Cyanotoxins in Lake Garda? The critical evaluation of chemodiversity. Workshop of Quality and sustainable use of water resources in Lake Garda and in other large water bodies in Europe: experiences within the project EULAKES. April 7-10, 2014, Palermo, Italy Capelli, C., Shams, S., Cerasino, L., Cavalieri, D., Papini, A., Salmaso, N. Toxin-producing cyanobacteria in the large lakes south of the Alps: detection of new producers and molecular identification methods. 10th PhD students meeting in Ecology and Aquatic Systems. April 7-12, 2013, Vienna, Austria Cerasino, L., Shams, S., Salmaso, N., Dietrich, D.R. The impact of toxic cyanobacteria on the water quality in the Deep Subalpine Lakes (DSL). European Geosciences Union General Assembly. June 26-July 1, 2011, Girona, Spain Ranjan, J., Shams, S., Cerasino, L., Salmaso, N. Ecological changes of central European lakes within the EU project EULAKES: nuisance cyanobacteria and impact on the aquatic ecosystems. The 7th Symposium of the European Freshwater Science. E. Research awards and scholarship March-May 2013 COST action award, Short Term Scientific Missions (STSMs) for learning new molecular methods at the laboratory of Prof. Kaarina Sivonen, Microbiology department, Helsinki University, Finland. October 29-November 2, 2012 Student Travel award for participation in 15th International Conference on Harmful Algae held in Changwon, Republic of Korea. January 2011-January 2015 PhD scholarship from Fondazione Edmund Mach, San Michele All'adige Trento, Italy. 7 II. Deusche Zusammenfassung Massive Blaualgen (Cyanobakterien)-blüten werden weltweit und zunehmend in Oberflächengewässern registriert. Diese Cyanobacterienblüten sind vielfach toxisch aufgrund der Präsenz von hepato- und neurotoxinen. Mehrere Vergiftungsfälle mit Menschen und Nutztieren aber auch Wildtieren sind auf diese toxischen Cyanobakterienblüten zurückzuführen. Entsprechend müssen Oberflächengewässer routinemässig auf das Vorkommen von Cyanobakterienblüten und möglicher Toxien untersucht werden. Im ersten Teil dieser Doktorarbeit wurden LC-MS Methoden zur Identifizierung und Quantifizierung von Cyanobakterientoxinen im Lago di Garda etabliert und optimiert. Anatoxin-a (ATX) und Microcystine (MCs) konnten im Lago di Garda regelmässig nachgewiesen werden, jedoch zu unterschiedlichen Jahreszeiten. ATX dominierte primär im Frühsommer während MCs typischerweise erst im Spätsommer/Spätherbst in höheren Konzentrationen nachzuweisen war. Obwohl 5 verschiedene MCs nachgewiesen werden konnten, dominierte das MC-RRdm Kongener in allen analysierten Proben. In einem weiteren Kapitel dieser Doktorarbeit wurden die kinetischen Aspekte des trophischen Transfers von MC in Planktothrix rubescens zum Wasserfloh Daphnia magna untersucht. Modelierungen der MC Akkumulation in der Wasserfloh Daphnia magna untersucht zeigetn, dass die MC Akkumulation prinzipiell von der ursprünglichen MC Konzentration und der Expositionsdauer der Daphnien abhängt. Innerhalb der ersten 24 h der Exposition ist die MC Akkumulation in der Wasserfloh, Daphnia magna, nahezu linear, unabhängig davon wie hoch die Dichte der Planktothrix rubescens und die MC Konzentration war. Nach 48h Exposition, verlief die MC Akkumulation in der Wasserfloh Daphnia magna exponentiell. Im letzten Teil dieser Doktorarbeit wurde ein polyphasischer Ansatz gewählt um die Oscillatoriales zu identifizieren, welche für die Synthese von ATX im Lago di Garda verantwortlich ist. Dieser Ansatz beinhaltete mikroskopische, molkelar-biologische und analytische Methoden. Aufgrund dieser Analysen konnte Tychonema bourrellyi als neuer ATX Produzent und auch die entsprechenden