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The present work describes the optimizationof a short-term assay, based on the inhibition ofthe esterase activity of the alga Pseudokirchneriellasubcapitata, in a microplate format. The optimizationof the staining procedure showed that the incubationof the algal cells with 20 μmolL−1 fluorescein diacetate(FDA) for 40 min allowed discrimination betweenmetabolic active and inactive cells. The shorttermassay was tested using Cu as toxicant. For thispurpose, algal cells, in the exponential or stationaryphase of growth, were exposed to the heavy metal ingrowing conditions. After 3 or 6 h, cells were subsequentlystained with FDA, using the optimized procedure.For Cu, the 3- and 6-h EC50 values, based on theinhibition of the esterase activity of algal cells in theexponential phase of growth, were 209 and 130 μgL−1, respectively. P. subcapitata cells, in the stationaryphase of growth, displayed higher effective concentrationvalues than those observed in the exponential phase. The 3- and 6-h EC50 values for Cu, for cellsin the stationary phase, were 443 and 268 μgL−1,respectively. This short-term microplate assay showedto be a rapid endpoint for testing toxicity using thealga P. subcapitata. The small volume required, thesimplicity of the assay (no washing steps), and theautomatic reading of the fluorescence make the assayparticularly well suited for the evaluation of the toxicityof a high number of environmental samples

Optimization of a Microplate-Based Assay to Assess Esterase Activity in the Alga Pseudokirchneriella subcapitata