Effects of Algae Frozen at Different Temperatures on Chronic Assessment Endpoints Observed withDaphnia magna

TheDaphnia magna21-day juvenile production test is not yet fully standardized because of many sources of variation. One is the diet provided to daphnids: the ration must be sufficient and the quality of algal cells must be optimal for achieving the required number of offspring defined by the new OECD guidelines. The experiments reported herein first examined the effects ofRaphidocelis subcapitataafter it had been maintained under four different conditions of storage (4, −20, −80, and −196°C) on the survival, reproductive performance (over 21 days), and growth (ascertained by dry weight) of individually heldD. magnafor three generations. Under all of the four regimes tested, daphnids survived and reproduced in a manner which fulfilled the current OECD guidelines for a valid test, but the best results were obtained with fresh algae and algae frozen at −80°C. Second, althoughD. magnahas been widely used to determine toxicity of chemical substances, there are no reports in the literature that describe a rigorous study of the nutritional quality of the algae given to daphnids. Therefore, cell number, optical density, amount of organic carbon, and esterasic activity (assessed by intracellular breakdown of FDA to fluorescein) of algae that have been preserved at 4, −20, −80, and −196°C were investigated. This part of the study indicated that freezing had no effect on cell numbers, in contrast to optical density, amount of organic carbon, and esterasic activity. First, it was found that esterasic activity was closely correlated to the reproductive performance of daphnids. It appears important, therefore, to consider the inclusion of this enzymatic activity as part of the routine quality control given to this microinvertebrate chronic procedure.