High resolution chlorophyll- a in-situ fluorescence sensors versus in-vitro chlorophyll- a measurements in mesocosms with contrasting nutrient and temperature treatments.

ABSTRACT

Harmful algal blooms (HABs) are a significant threat to freshwater ecosystems, and monitoring for changes in biomass is therefore important. Fluorescence in-situ sensors enable rapid and high frequency real-time data collection and have been widely used to determine chlorophyll- a (Chla) concentrations that are used as an indicator of the total algal biomass. However, conversion of fluorescence to equivalent Chla concentrations is often complicated due to biofouling, phytoplankton composition and the type of equipment used. Here, we validated measurements from 24 Chla and 12 phycocyanin (cyanobacteria indicator) fluorescence in-situ sensors (Cyclops-7F, Turner Designs) against spectrophotometrically (in-vitro) determined Chla and tested a data-cleaning procedure for eliminating data errors and impacts of non-photochemical quenching. The test was done across a range of freshwater plankton communities in 24 mesocosms (i.e. experimental tanks) with a 2x3 (high and low nutrient x ambient, IPCC-A2 and IPCC-A2+50% temperature scenarios) factorial design. For most mesocosms (tanks), we found accurate (r 2 ≥ 0.7) calibration of in-situ Chla fluorescence data using simple linear regression. An exception was tanks with high in-situ phycocyanin fluorescence, for which multiple regressions were employed, which increased the explained variance by >16%. Another exception was the low Chla concentration tanks (r 2 < 0.3). Our results also show that the high frequency in-situ fluorescence data recorded the timing of sudden Chla variations, while less frequent in-vitro sampling sometimes missed these or, when recorded, the duration of changes was inaccurately determined. Fluorescence in-situ sensors are particularly useful to detect and quantify sudden phytoplankton biomass variations through high frequency measurements, especially when using appropriate data-cleaning methods and accounting for factors that can impact the fluorescence readings.

Harmful algal blooms (HABs) may pose a significant threat to freshwater ecosystems and to animal and human health. Therefore, it is important to monitor changes in algal biomass. Traditional methods, while effective, lack the ability to provide rapid, high-frequency, real-time data. In-situ fluorescence sensors, specifically designed to measure chlorophyll- a (total phytoplankton indicator) and phycocyanin (Blue-green algae indicator), offer a promising solution. However, challenges such as biofouling, temporal changes in phytoplankton composition, and equipment variations complicate the conversion of fluorescence data into equivalent chlorophyll- a concentrations. Our study aimed to validate measurements from 24 chlorophyll- a and 12 phycocyanin fluorescence in-situ sensors (Cyclops-7F, Turner Designs). We compared these measurements against spectrophotometrically determined (in-vitro method) chlorophyll- a concentrations. Additionally, we tested a data-cleaning procedure to eliminate errors caused by different sources, such as light. The validation and testing were conducted at Lemming Experimental Mesocosm site (Denmark), in 24 experimental tanks (mesocosms) representing 2 different nutrient levels and 3 temperature scenarios. This study underlines that high-frequency in-situ fluorescence sensors can be useful, only if the user is aware of the possible interacting factors that can influence fluorescence readings (e.g. turbidity, daylight). Therefore, in-situ fluorescence sensors, when properly calibrated and validated, offer a valuable tool for monitoring harmful algal blooms. The high-frequency data provides insights into sudden variations in phytoplankton biomass, demonstrating the potential for improved real-time understanding of freshwater ecosystems.