Evaluating the estrogenicity of an effluent-dominated river in California, USA: Comparisons of in vitro and in vivo bioassays.

Estrogenically active compounds (EACs) in surface waters can disrupt the endocrine system of biota, raising concern for aquatic species. Concentrations of EACs are generally higher in effluent-dominated aquatic systems, such as California's Santa Ana River (SAR). Addressing estrogenicity of effluent-dominated waters is increasingly important due to both increasing urbanization and climate change. To this end, water samples were collected from multiple sites downstream of wastewater treatment plants (WWTPs) and intermittent points along the SAR during 2018-2019 and cell-based bioassays were used to determine estrogen receptor activity. During baseflow conditions, the highest estradiol equivalencies (EEQs) from all SAR water between summer (August and September) and fall (November) sampling events in 2018 were from Yorba Linda (EEQ = 1.36 ± 0.38 ng/L) and Prado (1.14 ± 0.13 ng/L), respectively. Water extracts in January 2019 following a major rainfall generally had higher EEQs with the highest EEQ of 10.0 ± 0.69 ng/L observed at Yorba Linda. During low flow conditions in November 2018, male Japanese medaka (Oryzias latipes) fish were exposed to SAR water to compare to cell bioassay responses and targeted analytical chemistry for 5 steroidal estrogens. Chemical-based EEQ correlations with in vitro EEQs were statistically significant. However, vitellogenin (vtg) mRNA expression in the livers of medaka exposed to SAR water was not significantly different compared to controls. These results indicate that seasonal variation and surface water runoff events influence estrogenic activity in the SAR and may induce estrogenic effects to native fish populations in wastewater-dominated streams in general.

Bioassay guided analysis coupled with non-target chemical screening in polyethylene plastic shopping bag fragments after exposure to simulated gastric juice of Fish.

In this study, fragments of polyethylene plastic bags were treated with simulated gastric juice of fish for 16 h. Following solid-phase extraction, methanol eluents caused acute toxicity to embryos and larvae of Japanese medaka. Chromatographic fractions (polar to more non-polar with numbers increasing) of the extract were evaluated for toxicity and estrogenic activity using medaka and an estrogen receptor (ER) cell-line. Fractions 6 and 9 had the highest estrogenic effects with relative hydrophobic chemicals. The vtg expression in fraction 6 was 22-fold higher than control, and the ER cellular response in fraction 9 was 8.5-fold higher than controls. Following non-target screening (NTS), several novel phthalates and phenols were identified in the above two fractions. Fractions 1 and 2 appeared to be primarily responsible for the acute toxicity observed with the whole extract. The hatching rate decreased to 36 % in fraction 2, and was not observed following exposure to fraction 1. NTS of these fractions indicated 635 and 808 entities, respectively, most without toxicity information. These results indicate plastic leachates from gastric juices of fish are complex mixtures of many compounds that can have acute reproductive and sublethal endocrine impacts in fish.