Factors affecting the growth of Pseudokirchneriella subcapitata in single-species tests: lessons for the experimental design and the reproducibility of a multi-trophic laboratory microcosm.

The need for an integrated risk assessment at an ecologically relevant scale (e.g., at the population/community levels) has been acknowledged. Multispecies systems with increased ecological complexity, however, are difficult if not impossible to reproduce. The laboratory-scale microcosm TriCosm (Pseudokirchneriella subcapitata, Ceriodaphnia dubia, Hydra viridissima) of intermediate complexity was developed for the reproducible assessment of chemical effects at the population/community levels. The system dynamics were repeatable in the short term, but interexperimental variation of algal dynamics in the long term triggered knock-on effects on grazer and predator populations. We present 20 experiments to assess the effects of 12 factors (test medium, vessel type/condition, shaking speed, light intensity/regime, inoculation density, medium preparation components, metal concentration/composition, buffering salt type/concentration) on algal growth in the TriCosm enclosure. Growth rates varied between <= 0 and 1.40 (+/- 0.21) and generally were greatest with increased shaking speed, light exposure, medium buffer, or aeration time. Treatments conducted in dishes with aseptically prepared, lightly buffered, and/or hardly aerated medium resulted in low growth rates. We found that inter-experimental variation of algal dynamics in the TriCosm was caused by a modification of medium preparation (omission of medium aeration) with the aim of reducing microbial contamination. Our findings highlight the facts that consistency in experimental procedures and in-depth understanding of system components are indispensable to achieve repeatability. Environ Toxicol Chem 2019;00:1-13. (c) 2019 SETAC