Automated color correction method for Vibrio fischeri toxicity test. Comparison of standard and kinetic assays.
We demonstrate in this study that the toxicity of solid and highly colorful samples can be measured with kinetic bioassay using luminescent bacterium Vibrio fischeri. The Flash assay, named after the test protocol, is performed with a tube luminometer. In this method, each sample acts as a reference for itself, and therefore, the color correction is possible with minimal hands-on-time. The bacteria are dispensed into the sample and the signal is recorded continuously. The maximum signal received after immediately dispensing is compared to the signal after an incubation period. With many chemicals, the toxic effects are obtained after a very short contact time. However, different chemicals have different modes of toxicity. Thus, kinetic data from sample analyses after 15 or 30 min for this bacterium gives an additional dimension for obtaining reliable results. The performance of the test was compared to the standardized photobacteria test protocol with reference chemicals. The repeatability of the test was excellent. The coefficient of variation was normally below 1% with 10 replicates.