Chlorophyll and phycocyanin in-situ fluorescence in mixed cyanobacterial species assemblages: Effects of morphology, cell size and growth phase

Cyanobacteria harmful blooms can represent a major risk for public health due to potential release of toxins and other noxious compounds in the water. A continuous and high-resolution monitoring of the cyanobacteria population is required due to their rapid dynamics, which has been increasingly done using in-situ fluorescence of phycocyanin (f-PC) and chlorophyll a (f-Chl a). Appropriate in-situ fluorometers calibration is essential because f-PC and f-Chl a are affected by biotic and abiotic factors, including species composition. Measurement of f-PC and f-Chl a in mixed species assemblages during different growth phases - representative of most field conditions - has received little attention. We hypothesized that f-PC and f-Chl a of mixed assemblages of cyanobacteria may be accurately estimated if taxa composition and fluorescence characteristics are known. We also hypothesized that species with different morphologies would have different fluorescence per unit cell and biomass. We tested these hypotheses in a controlled culture experiment in which photosynthetic pigment fluorescence, chemical pigment extraction, optical density and microscopic enumeration of four common cyanobacteria species (Aphanocapsa sp, Microcystis aeruginosa, Dolichospermum circinale and Raphidiopsis raciborskii) were quantified. Both monocultures and mixed cultures were monitored from exponential to late stationary growth phases. The sum of fluorescence of individual species calculated for mixed samples was not significantly different than measured fluorescence of mixed cultures. Estimated and measured f-PC and f-Chl a of mixed cultures had higher correlations and smaller absolute median errors when estimations were based on fluorescence per biomass instead of fluorescence per cell. Largest errors were overestimations of measured fluorescence for species with different morphologies. Fluorescence per cell was significantly different among most species, while fluorescence per unit biomass was not, indicating that conversion of fluorescence to biomass reduces species-specific bias. This study presents new information on the effect of species composition on cyanobacteria fluorescence. Best practices of deployment and operation of fluorometers, and data-driven models supporting in-situ fluorometers calibration are discussed as suitable solutions to minimize taxa-specific bias in fluorescence estimates.