Phaeodactylum tricornutum was exposed to various toxic substances (zinc, copper or dodecylbenzenesulfonic acid sodium salt) in accordance with the AlgalToxkit® protocol based on the UNI EN ISO 10253 method in order to quantitatively compare the responses obtained by traditional growth-rate inhibition tests with morphological (biovolume) and physiological (chlorophyll-a, phaeophytin ratio) endpoints. A novel approach is proposed for detecting early and sub-lethal effects based on biovolume quantification using confocal microscopy coupled with an image analysis system. The results showed that effects on both biovolume and the photosynthetic complex are sensitive and powerful early warning tools for evaluating sub-lethal effects of exposure. Specifically, biovolume showed significant sensitive and early responses for the tested surfactant. Qualitatively, we also observed structural anomalies and effects on natural auto-fluorescence in exposed cells that also represent potentially useful tools for ecotoxicological studies.
