Validation of Microplate Bioassays for the Assessment of Contaminated and Remediated Sites

Background, Goal and Scope  Bioassays are frequently used to investigate the water extractable ecotoxicological and genotoxicological potential of contaminated soil samples. A laboratory intercomparison study was performed for validation of miniaturised biological test systems for the assessment of contaminated and remediated sites. The successful performance of this study resulted in an optimisation of microplate assays with respect to the testing of chemicals and environmental samples. Methods  For this purpose, miniaturised bioassays were chosen, which, because of their stage of development, are suitable for routine application in the characterisation of the water extractable ecotoxicological and genotoxicological potential of soils. These ecotoxicological and genotoxicological assays were performed with contaminated soil samples by three institutions at the same time. Results and Discussion  The toxicological assessment of the contaminated and remediated soil samples using LID-values, as a rule, was highly uniform. Some minor deviations could, for the most part, be explained by the heterogeneity of the soil samples and, to a lesser extent, by methodical aspects. The difference in sensitivity towards contaminants of the two bacteria Vibrio fischeri and Pseudomonas putida was pointed out. In the algae test with Desmodesmus subspicatus, the influence of the highest sample concentrations on the growth controls became obvious. It was recommended to modify the experimental setup of the microtitration plate, i.e. to place growth controls located next to both the lowest and the highest dilution steps of the sample. The Ames-test did in some cases provide new information on the genotoxicity of the samples, but is not considered useful in a test battery for the evaluation of the genotoxic potential because of its great expense in time and work. Conclusions  The investigations in this laboratory-intercomparison study for the assessment of the water extractable toxic potential of soil samples show that different bioassays are needed, which, in contrast to chemical-analytical methods, can detect the complete effects of all present pollutants in contaminated and remediated soils and solid substrates path-specifically. Recommendations and Outlook  If the recommended modifications for the performance of the bacterial and algae growth inhibition assays on microplates are taken into consideration, these tests can substitute the tests performed on a macro scale. The usefulness of the umu-test and the NM2009-test for the investigation of the genotoxic potential has been proven. Although the tests performed on microplates require much lower sample amounts, it is recommended that sample amounts be eluted in accordance with current guidelines to ensure representativity of the sample. Further work should focus on toxicity identification studies in the future by combining chemical and toxicological analyses.