A Multi-Generation Sublethal Assay of Phenols Using the Nematode Caenorhabditis elegans

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Abstract
The nematode Caenorhabditis elegans (C. elegans) was used for a multiple-generation toxicity bioassay of phenols. We examined the sublethal toxicity (fecundity and reproduction) of bisphenol A (BisA), 4-(1-adamantyl) phenol (Adp), and 4,4′-(1,3-adamantanediyl) diphenol (AdDP) over five generations using a Nematode Growth Medium (NGM) 1.7% agar plate. In the fourth generation, the phenols affected the fecundity rate of C. elegans at doses 100- to 10000-fold lower than the LC50s. In particular, at 1 nM BisA, the number of worms decreased significantly to about 50% of control (p < 0.05). A comparison of the number of viable worms and eggs suggested that the phenols exert hatching toxicity. In addition, individuals with an abnormality in the vulva, which could not ovulate, appeared on the phenol-containing plates. We confirmed that phenols disrupt reproduction in C. elegans and that the method using NGM agar plates facilitates multi-generational toxicity tests of chemicals that are poorly soluble in water