| Plastic production and usage have increased, leading to an increase in plastic waste and pollution. Plastics contain additives, such as bisphenols, which can leach out, especially from smaller particles. BPA, a common bisphenol, is banned because of its endocrine-disrupting properties; however, substitutes such as BPE, BPAP, and BPZ may be equally or more toxic. This study compared the developmental toxicity profiles of BPA and its substitutes using an automated, unbiased, quantitative, and multi-endpoint zebrafish high content embryo test. The zebrafish embryos were exposed to various concentrations of BPA, BPZ, BPAP, BPE, and BPG under controlled conditions. Observations at 24- and 96-hours post fertilization (hpf) were used to assess mortality, spontaneous tail coiling (STC), photomotor response (PMR), hatching, and locomotor response (LMR). Morphological assessments were conducted using an Automated Imaging Robot (AIR). The analysis utilized several software tools for video and image processing, data extraction, and comprehensive data analysis. The data were further analyzed using KNIME workflows and R scripts. This study assessed dose-response relationships and effect concentrations (EC50) for lethality, behavior, and morphology in zebrafish embryos exposed to various chemicals, including bisphenols. The baseline toxicity was predicted using a specific QSAR model for zebrafish embryos. The comparative assessment was conducted by comparing the effect concentrations to baseline toxicity and observed mortality. Cluster analysis on effect ratios of bisphenols was also performed to compare toxicological profiles to Bisphenol A (BPA). Results indicate that these bisphenols have the potential to cause neurological, cardiovascular, and morphological effects. Generally, the toxicity profile of bisphenols showed that BPE was more toxic than the other bisphenols, both in terms of the number of affected endpoints and specificity. This study also found that BPAP and BPZ shared similar toxicity profiles, but were not identical to BPA. BPG effects were found to in the range of the predicted baseline toxicity. The lack of swim bladder was the most sensitive endpoint for all of the tested bisphenols, especially for BPE, which had the highest sensitivity ratio. In conclusion, this study indicated that the substitutes of BPA have similar or pronounced effect profiles based on the endpoints measured in this study, such as developmental toxicity, behavioral responses, and cardiotoxicity. The findings of this study may be useful for regulatory assessment of these bisphenols. |