, Volume 741, Issue 1, pp 33–49 | Cite as

Using an online phycocyanin fluorescence probe for rapid monitoring of cyanobacteria in Macau freshwater reservoir

  • Yijun Kong
  • Inchio LouEmail author
  • Yiyong Zhang
  • Chong U. Lou
  • Kai Meng Mok


Monitoring of cyanobacteria and their toxins are traditionally conducted by cell counting, chlorophyll-a (chl-a) determination and cyanotoxin measurements, respectively. These methods are tedious, costly, time consuming, and insensitive to rapid changes in water quality and cyanobacterial abundance. We have applied and tested an online phycocyanin (PC) fluorescence probe for rapid monitoring of cyanobacteria in the Macau Storage Reservoir (MSR) that is experiencing cyanobacterial blooms. The relationships among cyanobacterial abundance, biovolume, cylindrospermopsin concentration, and PC fluorescence were analyzed using both laboratory and in-the-field studies. The performance of the probe was compared with traditional methods, and its advantages and limitations were assessed in pure and mixed cyanobacterial cultures in the laboratory. The proposed techniques successfully estimated the species including Microcystis and Cylindrospermopsis, two toxic species recently observed in the MSR. During February–November, 2010, the PC probe detected high correlations between PC and cell numbers (R 2 = 0.71). Unlike the chl-a content, which indicates only the total algal biomass, the PC pigment specifically indicates cyanobacteria. These results support the PC parameter as a reliable estimate of cyanobacterial cell number, especially in freshwater bodies where the phytoplankton community and structure are stable. Thus, the PC probe is potentially applicable to online monitoring of cyanobacteria.


Phycocyanin fluorescence probe Online measurement Cyanobacteria Freshwater reservoir 



We thank the technical staff at Macao Water Co. Ltd. for collecting water samples, counting cyanobacterial cells and measuring chlorophyll-a concentration. Financial support from the Fundo para o Desenvolvimento das Ciências e da Tecnologia (FDCT) (grant # FDCT/016/2011/A) and Research Committee at University of Macau are gratefully acknowledged.


  1. APHA, 2005. Standard Methods for the Examination of Water and Wastewater, 21st ed. American Public Health Association, American Water Works Association and Water Environment Federation, Washington, DC.Google Scholar
  2. Bastien, C., R. Cardin, E. Veilleux, C. Deblois, A. Warren & I. Laurion, 2011. Performance evaluation of phycocyanin probes for the monitoring of cyanobacteria. Journal of Environmental Monitoring 13: 110–118.PubMedCrossRefGoogle Scholar
  3. Beutler, M., K. H. Wiltshire, B. Meyer, C. Moldaenke, C. Lüring, M. Meyerhöfer, U.-P. Hansen & H. Dau, 2002. A fluorometric method for the differentiation of algal populations in vivo and in situ. Photosynthesis Research 72: 39–53.PubMedCrossRefGoogle Scholar
  4. Brient, L., M. Lengronne, E. Bertrand, D. Rolland, A. Sipel, D. Steinmann, I. Baudin, M. Legeas, B. L. Rouzic & M. Bormans, 2008. A phycocyanin probe as a tool for monitoring cyanobacteria in freshwater bodies. Journal of Environment Monitoring 10: 248–255.CrossRefGoogle Scholar
  5. Cagnard, O., I. Baudin, I. Lemoigne & K. Cartnick, 2006. Assessment of emerging optic sensors (fluoroprobes) for algae on-line monitoring. American Water Works Association—Water Quality Technology Conference, Denver, CO, USA.Google Scholar
  6. Carmichael, W. W. & J. S. An, 1999. Using an enzyme linked immunosorbent assay (ELISA) and a protein phosphatase inhibition assay (PPIA) for the detection of microcystins and nodularins. Natural Toxins 7(6): 377–385.PubMedCrossRefGoogle Scholar
  7. Codd, G. A., 2000. Cyanobacterial toxins, the perception of water quality, and the prioritization of eutrophication control. Ecological Engineering 16: 51–60.CrossRefGoogle Scholar
  8. Codd, G. A., L. F. Morrison & J. S. Metcalf, 2005. Cyanobacterial toxins: risk management for health protection. Toxicology and Applied Pharmacology 203(3): 264–272.PubMedCrossRefGoogle Scholar
  9. Falconer, I., J. Bartram, I. Chorus, T. Kuiper-Goodman, H. Utkilen, M. Burch & G. A. Codd, 1999. Safe levels and safe practices. In Chorus, I. & J. Bartram (eds), Toxic Cyanobacteria in Water, A Guide to Their Public Health Consequences, Monitoring and Management. Spon Press, London, UK: 161–182.Google Scholar
  10. Gregor, J. & B. Marsalek, 2005. A simple in vivo fluorescence method for the selective detection and quantification of freshwater cyanobacteria and eukaryotic algae. Acta Hydrochimica et Hydrobiologica 33: 142–148.Google Scholar
  11. Gregor, J., B. Maršálek & H. Šípková, 2007. Detection and estimation of potentially toxic cyanobacteria in raw water at the drinking water treatment plant by in vivo fluorescence method. Water Research 41: 228–234.PubMedCrossRefGoogle Scholar
  12. Hillebrand, H., C. D. Dürselen, D. Kirschtel, D. Pollingher & T. Zohary, 1999. Biovolume calculation for pelagic and benthic microalgae. Journal of Phycology 35: 403–424.CrossRefGoogle Scholar
  13. Izydorczyk, K., M. Tarczynska, T. Jurczak, J. Mrowczynski & Zalewski Ma, 2005. Measurement of phycocyanin fluorescence as an online early warning system for cyanobacteria in reservoir intake water. Environmental Toxicology 20: 425–430.PubMedCrossRefGoogle Scholar
  14. Izydorczyk, K., C. Carpentier, J. Mrówczyński, A. Wagenvoort, T. Jurczak & M. Tarczyńska, 2009. Establishment of an alert level framework for cyanobacteria in drinking water resources by using the algae online analyser for monitoring cyanobacterial chlorophyll a. Water Research 43: 989–996.PubMedCrossRefGoogle Scholar
  15. Kurmayer, R. & T. Kutzenberger, 2003. Application of real-time PCR for quantification of microcystin genotypes in a population of the toxic cyanobacterium Microcystis sp. Applied and Environmental Microbiology 69: 6723–6730.PubMedCrossRefPubMedCentralGoogle Scholar
  16. Lawton, L. A., C. Edwards & G. A. Codd, 1994. Extraction and high performance liquid chromatographic method for determination of microcystins in raw and treated waters. Analyst 119(7): 1525–1530.PubMedCrossRefGoogle Scholar
  17. Leboulanger, C., U. Dorigo, S. Jacquet, B. Le Berre, G. Paolini & J.-F. Humbert, 2002. Use of a submersible spectrofluorometer (FluoroProbe) for the survey of a toxic cyanobacteria, Planktothrix rubescens, in a large alpine lake. Aquatic Microbial Ecology 30: 83–89.CrossRefGoogle Scholar
  18. Long, B. M., G. J. Jones & P. T. Orr, 2001. Cellular microcystin content in N-limited Microcystis aeruginosa can be predicted from growth rate. Applied and Environmental Microbiology 67: 278–283.PubMedCrossRefPubMedCentralGoogle Scholar
  19. McAlice B. J., 1971. Phytoplankton sampling with Sedgwick-Rafter cell. Limnology and Oceanography 16: 19–28.Google Scholar
  20. Mcquaid, N., A. Zamyadi, M. Prévost, D. F. Bird & S. Dorner, 2011. Use of in vivo phycocyanin fluorescence to monitor potential microcystin—producing cyanobacteria bivolume in a drinking water source. Journal of Environmental Monitoring 13: 455–463.PubMedCrossRefGoogle Scholar
  21. Orr, P. T. & G. J. Jones, 1998. Relationship between microcystin production and cell division rates in nitrogen-limited Microcystis aeruginosa cultures. Limnology and Oceanography 43: 1604–1614.CrossRefGoogle Scholar
  22. Paerl, H. W. & J. Huisman, 2009. Climate change: a catalyst for global expansion of harmful cyanobacterial blooms. Environmental Microbiology 1: 27–37.CrossRefGoogle Scholar
  23. Parésys, G., C. Rigart, B. Rousseau, A. W. M. Wong, F. Fan, J.-P. Barbier & J. Lavaud, 2005. Quantitative and qualitative evaluation of phytoplankton communities by trichromatic chlorophyll fluorescence excitation with special focus on cyanobacteria. Water Research 39: 911–921.PubMedCrossRefGoogle Scholar
  24. Richardson, T. L., E. Lawrenz, J. L. Pinckney, R. C. Guajardo, E. A. Walker, H. W. Paerl & H. L. MacIntyre, 2010. Spectral fluorometric characterization of phytoplankton community composition using the algae online analyzer. Water Research 44: 2461–2472.PubMedCrossRefGoogle Scholar
  25. Seppala, J., P. Ylostalo, S. Kaitala, S. Hallfors, M. Raateoja & P. Maunula, 2007. Ship-of-opportunity based phycocyanin fluorescence monitoring of the filamentous cyanobacteria bloom dynamics in the Baltic Sea. Estuarine, Coastal and Shelf Science 73(3–4): 489–500.CrossRefGoogle Scholar
  26. WHO, 1998. Guidelines for Drinking-Water Quality, 2nd ed., Addendum to Vol. 2, Health Criteria and Other Supporting Information. World Health Organisation, Geneva.Google Scholar
  27. Zamyadi, A., 2011. PhD thesis. Ecole polytechnique de Montreal, University of Montreal.Google Scholar
  28. Zamyadi, A., N. McQuaid, M. Prévost & S. Dorner, 2012. Monitoring of potentially toxic cyanobacteria using an online multi-probe in drinking water sources. Journal of Environmental Monitoring 14: 579–588.PubMedCrossRefGoogle Scholar
  29. Zhang, W., I. Lou, W. K. Ung, Y. Kong & K. M. Mok, in press. Using quantitative real-time PCR to characterize spatial and temporal variations of cylindrospermopsin- and microcystin-producing genotypes, and cyanotoxins concentrations in Macau Storage Reservoir. Frontiers of Earth Science.Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Yijun Kong
    • 1
  • Inchio Lou
    • 2
    Email author
  • Yiyong Zhang
    • 1
  • Chong U. Lou
    • 1
  • Kai Meng Mok
    • 2
  1. 1.Laboratory & Research CenterMacao Water Co. Ltd.Macau SARChina
  2. 2.Department of Civil and Environmental Engineering, Faculty of Science and TechnologyUniversity of MacauMacau SARChina

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