A Plate Based Assay for Determination of the Median Lethal Dose of 1-Hydroxyphenazine in Caenorhabditis elegans

Caenorhabditis elegans is an ideal model organism for studying the xenobiotic detoxification pathways of various natural and synthetic toxins. We developed a new workflow to study the effects of 1-hydroxyphenazine (1-HP), a toxin produced by the bacterium Pseudomonas aeruginosa, on the survival of C. elegans. Prior research has demonstrated that C. elegans can detoxify 1-HP through the general mechanism of O-glycosylation. As part of the Vertically Integrated Projects (VIP) undergraduate research team, we have developed a workflow for a plate-based toxicity assay to determine the median lethal dose (LD50) of 1-HP. This was achieved through a toxin exposure method in which the worms were exposed to varying concentrations of 1-HP. The death rates were measured using a fluorescent bead assay. This workflow can be used to test C. elegans responses to different toxins and also the response of different mutant strains to a toxin of interest.