Thousands of organic micropollutants and their transformation products occur in water. Although often present at low concentrations, individual compounds contribute to mixture effects. Cell-based bioassays that target health-relevant biological endpoints may therefore complement chemical analysis for water quality assessment. The objective of this study was to evaluate cell-based bioassays for their suitability to benchmark water quality and to assess efficacy of water treatment processes. The selected bioassays cover relevant steps in the toxicity pathways including induction of xenobiotic metabolism, specific and reactive modes of toxic action, activation of adaptive stress response pathways and system responses. Twenty laboratories applied 103 unique in vitro bioassays to a common set of 10 water samples collected in Australia, including wastewater treatment plant effluent, two types of recycled water (reverse osmosis and ozonation/activated carbon filtration), stormwater, surface water, and drinking water. Sixty-five bioassays (63%) showed positive results in at least one sample, typically in wastewater treatment plant effluent, and only five (5%) were positive in the control (ultrapure water). Each water type had a characteristic bioanalytical profile with particular groups of toxicity pathways either consistently responsive or not responsive across test systems. The most responsive health-relevant endpoints were related to xenobiotic metabolism (pregnane X and aryl hydrocarbon receptors), hormone-mediated modes of action (mainly related to the estrogen, glucocorticoid, and antiandrogen activities), reactive modes of action (genotoxicity) and adaptive stress response pathway (oxidative stress response). This study has demonstrated that selected cell-based bioassays are suitable to benchmark water quality and it is recommended to use a purpose-tailored panel of bioassays for routine monitoring.

Organotin compounds released from antifouling paints, such as tributyltin (TBT) and triphenyltin (TPT), are potent inducers of imposex (a superimposition of male genital tracts, such as penis and vas deferens, on females) in marine gastropods. Little is known about the induction mechanism of gastropod imposex. Here, we show that organotins bind the human retinoid X receptors (hRXRs) with high affinity and that injection of 9-cis retinoic acid (RA), the natural ligand of hRXRs, into females of the rock shell (Thais clavigera) induces the development of imposex. Cloning of the RXR homologue from T. clavigera revealed that the ligand-binding domain of rock shell RXR was very similar to vertebrate RXR and bound to both 9-cis RA and to organotins. These suggest that RXR plays an important role in inducing the development of imposex, namely, the differentiation and growth of male genital tracts in female gastropods.

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High concentrations of bisphenol A were detected in the effluent from several pulping processes for waste paper containing thermal paper and/or other printed paper. Chlorinated derivatives of bisphenol A were found to be formed by its reaction with a low concentration of chlorine in the effluent from the bleaching process using sodium hypochlorite. Poly-chlorinated derivatives were mainly detected in the final effluents from two plants because they were not biodegraded in the water recycling process by treatment with activated sludge. The estrogenic activities of bisphenol A and its chlorinated derivatives were evaluated by an agonist assay using the yeast two-hybrid system with and without a metabolic activation test using rat liver S9. All of the chlorinated derivatives tested showed more potent activity than bisphenol A without S9. The activity of 3,3′-dichlorinated BPA was 38-fold stronger than that of bisphenol A. The activities of these compounds were almost eliminated upon treatment with S9.