Bioassays with unicellular algae: deviations from exponential growth and its implications for toxicity test results

Growth assays with unicellular green algae are an established tool in ecotoxicological effect assessment for chemicals and environmental samples. From an ecological perspective it seems appropriate to use the growth rate as a process variable rather than a measure of biomass gain for calculating inhibitory effects of contaminants. The notion of simple exponential growth for the description of the population increase in undisturbed suspension cultures of unicellular green algae, however, seems to be an oversimplification. Experimental findings describe the increase in biomass, cell number, the development of cell volume distributions of populations, and the relationship between cell size and chlorophyll content for individual cells over one generation at a time resolution of 2-h intervals. It was observed that algal populations of Desmodesmus subspicatus show a time pattern of cell size growth; the average cell volume increases about sixfold, without corresponding increase in population size. This is followed by a distinct cell division phase with little gain in biomass. This synchronous growth behavior despite continuous illumination may be explained by the multiple fission characteristic of unicellular green algae which is an adaptation to cyclic light-dark changes in the environment. It might be controlled by an independent cell cycle clock. For routine regulatory testing fluorescence-based measurements rather than cell counting minimizes the confounding effect on toxicity determination. For investigations of time-dependent effects, e.g., by pulsed exposure, an alternative mechanistically based growth function for unicellular algae is proposed that accommodates for the observed growth pattern.