Assessment of the relative cardiotoxicity and behavioral effects of butylated hydroxytoluene and its metabolites in developing zebrafish (Danio rerio).

Butylated hydroxytoluene (BHT) and its transformation products are ubiquitously detected in aquatic environments. Despite studies reporting on the adverse effects of BHT exposure in early-staged zebrafish, the comparative toxicity of its metabolites is not known. To address this, zebrafish embryos were exposed continuously to 0.01-1 μM BHTs (BHT, BHT-Q, BHT-OH, BHT-CHO, and BHT-COOH) for up to 6 days. Each chemical altered the heart rates of developing zebrafish at 72 hpf. When assessing cardiac morphology at 48 and 72 hpf, BHT-COOH showed stronger effects compared to BHT in inducing pericardial edema and myocardial hypertrophy. This effect is hypothesized to be attributed to differences in transcriptional regulation of key genes during cardiac development. BHT predominantly affected transcript expression in the early (15 hpf) or late stages (48-72 hpf) of heart development, but did not impact the transcription during the middle stage (24-36 hpf). Conversely, BHT-COOH altered the expression of early transcription factors (i.e. tbx5, nkx2.5, gata4, hand2) and functional genes (i.e. myh6, nppa, cacna1ab, and atp2a2) at all stages of cardiac development. The effects on the heart may also be related to behavioral changes observed in larval fish. Behavior was most sensitive with low chemical exposures (0.01 μM), while at higher concentrations, heart rate was a more sensitive indicator of BHT-induced toxicity. Taken together, our results indicate that BHTs induce adverse effects on cardiovascular function and BHT metabolites may pose higher risk than the parent compound.