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

Caroline M. Hodges, Susanna A. Wood, Jonathan Puddick, Christopher G. McBride, David P. Hamilton

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.