Study on Calibration Method for the Count of Living Algal Cells Detection Based on Variable Fluorescence in Ballast Water.

Abstract

In the monitoring the discharge of ballast water, the count of living algal cells is of utmost significant. Variable fluorescence, denoted as Fv, stands as an optimal parameter for photosynthetic fluorescence, efficiently charactering the living algal cells count, unaffected by the ballast waters' complex background fluorescence environment. This study deeply investigates the quantitative relationship between Fv and the count of living algal cells. Observations indicate that single cell fluorescence yield (abbreviated as SCF) varies significantly across different algae species, leading to considerable errors in quantifying living algal cell count in ballast water with unknown components using the calibration relationship between Fv and the cell count. Thus, correcting SCF prior to calibration becomes necessary. The paper proposes an innovative SCF correction method based on cell cross-sectional area and an eμ factor (where μ is the expected value of the functional absorption cross-section of PSII) This method mitigates the influence of cell size and species differences on quantifying the living algal cell count. Correction operation trials revealed that dividing the SCF measurement by cell cross-sectional area and multiplying by eμ enhanced the correction effect. Comparative experiments demonstrated marked improvement: Relative errors (REs) for Chlorella pyrenoidosa and Chlorella marine, both belonging to the Chlorophyta group, fell from 92.1% and 90.6% to 37.2% and 9.5% respectively post-correction. Similarly, REs for Thalassiosira weissflogii and Nitzschia closterium minutissima, from the Bacillariophyta group, decreased from 74.7% and 68.1% to 14.3% and 19.1% respectively. The RE of Peridinium from the Pyrrophyta group dropped from 28.4% to 12.1%. The results underscore the effectiveness of cell cross-sectional area and eμ in correcting SCF, thus offering a novel correction method for swift and precise measurement of living algal cell count in ballast water, based on variable fluorescence.