1 The Use of Pigment Pattern for Online Algae Monitoring Detlev Lohse bbe Moldaenke Kiel

2 n Photosynthesis and Chl-Fluorescence n Spectral Groups of Phytoplankton n Principle of Spectral Fingerprinting n Instruments to Determine Algae Topics

3 Blue-green algae (Cyano- bacteria) in inland waters Threats Planktothrix rubescens Toxins Taste Odour Microcystin LR β-methyl aminoalanine Geosmin 2-MIB

4 Cyanobacterial compounds Biotoxins have been implicated in causing irreversible hepatotoxicity and tumor promoting reactions in laboratory rats Evidence in China suggests a correlation between microcystins in drinking water and primary liver cancer may have a possible connection to degenerative nerve deseases (Alzheimer´s, Parkinson´s)

5  Drinking Water  Recreational Water Blue-green “algae” (Cyanobacteria) in inland waters WHO Guidelines µg Microcystin/L Scum formation cells cyanobacteria/ml 10 µg Chl_a/L (cyanobacteria)

6 Phytoplankton = free-swimming or free-floating photosynthetic microorganism Microplankton Nanoplankton Picoplankton

7 Location of Photosynthesis

8 kfkf kpkp ktkt IoIo PSII Electron transport chain Peripheral antenna Fluorescence (685 nm) Thermal decay Absorption of light Starch Photosystem Reactions

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10 Cyanophyta Spectral groups of phytoplankton

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12 The Excitation of Algae LEDs at different wavelengths excite the algal pigments

13 Composite sprectral data

14 Normspectra (Fingerprint)

15 Yellow Substances

16 Deconvolution of prepared algae mixtures

17 Set-up of submersible probe

18 The Algae Online Analyser Chlorophyll Determination n Chlorophyll Determination n Algae Class Differentiation n Yellow Substance Measurement n Activity of Algae n Transmission n Measuring Range: µg/l n Continuous Flow n Batch mode n Automatic Cleaning of Chamber

19 Graphic Display of the AOA (Algae Online Analyser)

20 Migration of Dinoflagellates in eutrophic Plussee 10 h14 h

21 Outstanding problems Colonies Turbidity

22 Project

23 Different Methods for Chlorophyll determination In vivo Photometry In vivo fluorescence Extraction of pigments wet chemical analysis (ISO) HPLC separation, photometry, fluorometry

24 Optical methods Photometry in vitro Fluorescence in vivo Fluorescence indirect indirectindirect sample preparation sample preparation no sample preparation delay delay real-time low sensitivity high sensitivity high sensitivity accuracy poor accuracy medium accuracy medium

25 Conclusion: the In Vivo algae Fluorescence assay combines rapid spectrofluorometric measurement with a data pool of normed curves obtained from cultured algae. A mathematical approach uses algorithms for algae differentiation. IVF enables correction for Yellow Substances.

26 Thank you