Environmental Science and Pollution Research

, Volume 25, Issue 2, pp 1079–1088 | Cite as

Sensor manufacturer, temperature, and cyanobacteria morphology affect phycocyanin fluorescence measurements

  • Caroline M. Hodges
  • Susanna A. Wood
  • Jonathan Puddick
  • Christopher G. McBride
  • David P. HamiltonEmail author
Research Article


Sensors to measure phycocyanin fluorescence in situ are becoming widely used as they may provide useful proxies for cyanobacterial biomass. In this study, we assessed five phycocyanin sensors from three different manufacturers. A combination of culture-based experiments and a 30–sample field study was used to examine the effect of temperature and cyanobacteria morphology on phycocyanin fluorescence. Phycocyanin fluorescence increased with decrease in temperature, although this varied with manufacturer and cyanobacterial density. Phycocyanin fluorescence and cyanobacterial biovolume were strongly correlated (R 2 > 0.83, P < 0.05) for single-celled and filamentous species. The relationship was generally weak for a colonial strain of Microcystis aeruginosa. The colonial culture was divided into different colony size classes and phycocyanin measured before and after manual disaggregation. No differences were measured, and the observation that fluorescence spiked when large colonial aggregates drifted past the light source suggests that sample heterogeneity, rather than lack of light penetration into the colonies, was the main cause of the poor relationship. Analysis of field samples showed a strong relationship between in situ phycocyanin fluorescence and spectrophotometrically measured phycocyanin (R 2 > 0.7, P < 0.001). However, there was only a weak relationship between phycocyanin fluorescence and cyanobacterial biovolume for two sensors (R 2 = 0.22–0.29, P < 0.001) and a non-significant relationship for the third sensor (R 2 = 0.29, P > 0.4). The five sensors tested in our study differed in their output of phycocyanin fluorescence, upper working limits (1200 to > 12,000 μg/L), and responses to temperature, highlighting the need for comprehensive sensor calibration and knowledge on the limitations of specific sensors prior to deployment.


Aphanizomenon Cyanobacterial blooms Environmental monitoring Dolichospermum Microcystis Nodularia 



The authors thank Hugo Borges, Konstanze Steiner, and Oonagh Daly for their assistance in the field and Andrew Mahon and Marc Jary (Cawthron Institute) for technical support.

Funding information

This research was supported by the New Zealand Ministry of Business, Innovation and Employment (UOWX1503; Enhancing the health and resilience of New Zealand lakes) and the Marsden Fund of the Royal Society of New Zealand (12-UOW-087; Toxic in crowds). CMH was funded by a Waikato University Masters Scholarship.

Supplementary material

11356_2017_473_MOESM1_ESM.docx (197 kb)
ESM 1 (DOCX 197 kb).


  1. Ahn C, Joung S, Oh H (2007) Alternative alert system for cyanobacterial bloom, using phycocyanin level as a determinant. J Microbiol 45:98–104Google Scholar
  2. Bastien C, Cardin R, Veilleux E, Deblois C, Warren A, Laurion I (2011) Performance evaluation of phycocyanin probes for the monitoring of cyanobacteria. J Environ Monit 13:110–118CrossRefGoogle Scholar
  3. Bennett A, Bogorad L (1973) Complementary chromatic adaptation in a filamentous blue-green alga. J Cell Biol 58:419–435CrossRefGoogle Scholar
  4. Beutler M, Wiltshire KH, Meyer B, Moldaenke C, Lüring C, Meyerhöfer M, Hansen UP, Dau H (2002) A fluorometric method for the differentiation of algal populations in vivo and in situ. Photosynth Res 72:39–53CrossRefGoogle Scholar
  5. Bolch CS, Blackburn S (1996) Isolation and purification of Australian isolates of the toxic cyanobacterium Microcystis aeruginosa Kutz. J Appl Phycol 8:5–13CrossRefGoogle Scholar
  6. Bowling L, Ryan D, Holliday J, Honeyman G (2012) Evaluation of in situ fluorometry to determine cyanobacterial abundance in the Murray and Lower Darling Rivers, Australia. River Res Appl 29:1059–1071Google Scholar
  7. Brient L, Lengronne M, Bertrand E, Rolland D, Sipel A, Steinmann D, Baudin I, Legeas M, Le Rouzie B, Bormans M (2008) A phycocyanin probe as a tool for monitoring cyanobacteria in freshwater bodies. J Environ Monit 10:248–255CrossRefGoogle Scholar
  8. Chang D-W, Hobson P, Burch M, Lin T-F (2012) Measurement of cyanobacteria using in-vivo fluoroscopy—effect of cyanobacterial species, pigments, and colonies. Water Res 46:5037–5048CrossRefGoogle Scholar
  9. Codd G, Morrison L, Metcalf J (2005) Cyanobacterial toxins: risk management for health protection. Toxicol Appl Pharmacol 203:264–272CrossRefGoogle Scholar
  10. Fallon RD, Brock TD (1979) Lytic organisms and photooxidative effects: influence on blue-green algae (cyanobacteria) in Lake Mendota, Wisconsin. Appl Environ Microbiol 38:499–505Google Scholar
  11. Gregor J, Marsalek B (2004) Freshwater phytoplankton quantification by chlorophyll a: a comparative study of in vitro, in vivo and in situ methods. Water Res 38:517–522CrossRefGoogle Scholar
  12. Gregor J, Marsalek B, Sipkova H (2007) Detection and estimation of potentially toxic cyanobacteria in raw water at the drinking water treatment plant by in vivo fluorescence method. Water Res 41:228–234CrossRefGoogle Scholar
  13. Guibalt GG (1990) General aspects of luminescence spectroscopy. In: Guibalt GG (ed) Practical Fluorescence, 2nd edn. Marcel Dekker Inc, New YorkGoogle Scholar
  14. Gumbo JE, Cloete TE (2011) Light and electron microscope assessment of the lytic activity of Bacillus on Microcystis aeruginosa. Afr J Biotechnol 10:8054–8063CrossRefGoogle Scholar
  15. Hamilton DP, Carey CC, Arvola L, Arzberger P, Brewer C, Cole JJ, Gaiser E, Hanson PC, Ibelings BW, Jennings E, Kratz TK, Lin F-P, McBride CG, de Motta MD, Muraoka K, Nishri A, Qin B, Read JS, Rose KC, Ryder E, Weathers KC, Zhu G, Trolle D, Brookes JD (2014) A global lake ecological observatory network (GLEON) for synthesising high-frequency sensor data for validation of deterministic ecological models. Inland Waters 5:49–56CrossRefGoogle Scholar
  16. Harke M, Steffen M, Gobler C, Otten T, Wilhelm S, Wood SA, Pearl H (2016) A review of the global ecology, genomics, and biogeography of the toxic cyanobacterium, Microcystis. Harmful Algae 54:4–20CrossRefGoogle Scholar
  17. Hötzel G, Croome R (1999) A phytoplankton methods manual for Australian freshwaters. LWRRDC Occasional Paper 22/99. Land and Water Research and Development Corporation: CanberraGoogle Scholar
  18. Izydorczyk K, Tarczynska M, Jurczak T, Mrowczynski J, Zalewski M (2005) Measurement of phycocyanin and fluorescence as an online early warning system for cyanobacteria in reservoir intake water. Environ Toxicol 20:425–430CrossRefGoogle Scholar
  19. Kasinak JE, Holt BM, Chislock MF, Wilson AE (2015) Benchtop fluorometry of phycocyanin as a rapid approach for estimating cyanobacterial biovolume. J Plankton Res 37:248–257Google Scholar
  20. Kong Y, Lou I, Zhang Y, Lou CU, Mok KM (2014) Using an online phycocyanin fluorescence probe for rapid monitoring of cyanobacteria in Macau freshwater reservoir. Hydrobiologia 741:33–49CrossRefGoogle Scholar
  21. Leboulanger C, Dorigo U, Jacquet S, Le Berre B, Paolini G, Humbert JF (2002) Application of a submersible spectrofluorometer for rapid monitoring of freshwater cyanobacterial blooms: a case study. Aquat Microb Ecol 30:83–89CrossRefGoogle Scholar
  22. McQuaid N, Zamyadi A, Prevost M, Bird DF, Dorner S (2011) Use of in vivo phycocyanin fluorescence to monitor potential microcystin-producing cyanobacterial biovolume in a drinking water source. J Environ Monit 13:455–463CrossRefGoogle Scholar
  23. Olenina I, Hadju S, Edler L, Anderson S, Wasmund N, Busch S, Gobel J, Gromisz S, Huseby S, Huttunen M, Jaanus A, Kokkonen P, Ledaine I, Niemkiewicz E (2006) Biovolumes and size-classes of phytoplankton in the Baltic Sea. HELCOM Balt Sea Environ Proc No. 106, pp 144Google Scholar
  24. Puddick J, Prinsep MR, Wood SA, Kaufononga SA, Cary SC, Hamilton DP (2014) High levels of structural diversity observed in microcystins from Microcystis CAWBG11 and characterization of six new microcystin congeners. Marine Drugs 12:5372–5395CrossRefGoogle Scholar
  25. Rhodes L, Smith K, Wood SA, Ponikla K, MacKenzie L, Harwood T, Munday R (2015) A significant national collection—the Cawthron Institute Culture Collection of Micro-Algae. N Z J Mar Freshwat Res 50:291–316CrossRefGoogle Scholar
  26. Song K, Li L, Tedesco L, Clercin N, Hall B, Li S, Liu D, Sun Y (2013) Remote estimation of phycocyanin (PC) for inland waters coupled with YSI PC fluorescence probe. Environ Sci Poll Res 20:5330–5340CrossRefGoogle Scholar
  27. Turner Designs (2015) Technical note: an introduction to fluorescence measurements. Retrieved from
  28. Wood SA, Hamilton DP, Paul WJ, Safi KA, Williamson WM (2009) New Zealand guidelines for managing cyanobacteria in recreational fresh waters—interim guidelines. Ministry for the Environment, WellingtonGoogle Scholar
  29. Wood SA, Dietrich DR, Cary SC, Hamilton DP (2012) Increasing Microcystis cell abundance enhances microcystin synthesis: a mesocosm study. Inland Waters 2:17–22CrossRefGoogle Scholar
  30. Wood SA, Borges H, Puddick J, Biessy L, Atalah J, Hawes I, Dietrich D, Hamilton DP (2016) Contrasting cyanobacterial communities and microcystin concentrations in summers with extreme weather events: insights into potential effects of climate change. Hydrobiologia 785:71–89CrossRefGoogle Scholar
  31. Zamyadi A, McQuaid N, Prévost M, Dorner S (2012) Monitoring of potentially toxic cyanobacteria using an online multi-probe in drinking water sources. J Environ Monit 14:579–588CrossRefGoogle Scholar
  32. Zamyadi A, Choo F, Newcombe G, Stuetz R, Henderson RK (2016) A review of monitoring technologies for real-time management of cyanobacteria: recent advances and future direction. Trends Anal Chem 85:83–96CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  1. 1.Environmental Research InstituteUniversity of WaikatoHamiltonNew Zealand
  2. 2.Cawthron InstituteNelsonNew Zealand
  3. 3.Australian Rivers InstituteGriffith UniversityBrisbaneAustralia

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