A Comparison of In Vitro Metabolic Clearance of Various Regulatory Fish Species Using Hepatic S9 Fractions.

Bioaccumulation predictions can be substantially improved by combining in vitro metabolic rate measurements derived from rainbow trout hepatocytes and/or hepatic S9 fractions with quantitative structure-activity relationship (QSAR) modeling approaches. Compared with in vivo testing guidelines Organisation for Economic Co-operation and Development (OECD) 305 and Office of Chemical Safety and Pollution Prevention (OCSPP; an office of the US Environmental Protection Agency) 850.1730, the recently adopted OECD test guidelines 319A and 319B are in vitro approaches that have the potential to provide a time- and cost-efficient, humane solution, reducing animal use while addressing uncertainties in bioaccumulation across species. The present study compares the hepatic clearance of the S9 subcellular fraction of rainbow trout, bluegill, common carp, fathead minnow, and largemouth bass, discerning potential differences in metabolism between different warm- and cold-water species. With refinements to the in vitro metabolic S9 assay for high-throughput analysis, we measured in vitro clearance rates of seven chemicals crossing multiple classes of chemistry and modes of action. We confirmed that data from rainbow trout liver S9 fraction metabolic rates can be utilized to predict rainbow trout bioconcentration factors using an in vitro to in vivo extrapolation model, as intended in the OECD 319B applicability domain per the bioaccumulation prediction. Also, we determined that OECD 319B can be applied to other species, modified according to their habitat, adaptations to feeding behavior, and environmental conditions (e.g., temperature). Once toxicokinetics for each species is better understood and appropriate models are developed, this method can be an excellent tool to determine hepatic clearance and potential bioaccumulation across species. The present study could be leveraged prior to or in place of initiating in vivo bioconcentration studies, thus optimizing selection of appropriate fish species. Environ Toxicol Chem 2024;43:1390-1405. © 2024 SETAC.

Arsenic exposure induces a bimodal toxicity response in zebrafish.

In toxicology, standard sigmoidal concentration-response curves are used to predict effects concentrations and set chemical regulations. However, current literature also establishes the existence of complex, bimodal concentration-response curves, as is the case for arsenic toxicity. This bimodal response has been observed at the molecular level, but not characterized at the whole organism level. This study investigated the effect of arsenic (sodium arsenite) on post-gastrulated zebrafish embryos and elucidated effects of bimodal concentration-responses on different phenotypic perturbations. Six hour post fertilized (hpf) zebrafish embryos were exposed to arsenic to 96 hpf. Hatching success, mortality, and morphometric endpoints were evaluated both in embryos with chorions and dechorionated embryos. Zebrafish embryos exhibited a bimodal response to arsenic exposure. Concentration-response curves for exposed embryos with intact chorions had an initial peak in mortality (88%) at 1.33 mM arsenic, followed by a decrease in toxicity (~20% mortality) at 1.75 mM, and subsequently peaked to 100% mortality at higher concentrations. To account for the bimodal response, two distinct concentration-response curves were generated with estimated LC10 values (and 95% CI) of 0.462 (0.415, 0.508) mM and 1.69 (1.58, 1.78) mM for the 'low concentration' and 'high concentration' peaks, respectively. Other phenotypic analyses, including embryo length, yolk and pericardial edema all produced similar concentration-response patterns. Tests with dechorionated embryos also resulted in a bimodal toxicity response but with lower LC10 values of 0.170 (0.120, 0.220) mM and 0.800 (0.60, 0842) mM, respectively. Similarities in bimodal concentration-responses between with-chorion and dechorionated embryos indicate that the observed effect was not caused by the chorion limiting arsenic availability, thus lending support to other studies such as those that hypothesized a conserved bimodal mechanism of arsenic interference with nuclear receptor activation.

Correlating Quantitative Measurements of Radical Production by Photocatalytic TiO2 with Daphnia magna Toxicity.

Increased use of titanium dioxide (TiO2 ) nanoparticles (NPs) in domestic and industrial applications has increased the risk for adverse environmental outcomes based on an elevated likelihood of organism exposure. Anatase TiO2 NP exposure to ultraviolet A (UV-A) radiation in aquatic environments generates radical oxygen species (ROS), which may ultimately be responsible for increased organism toxicity. We have identified and measured the 2 most relevant ROS species, hydroxyl and superoxide radicals, and described that ROS can be modeled using the highly reactive hydroxyl radical to provide an upper bound for toxicity. The TiO2 NPs were co-exposed to increasing natural organic matter (NOM) amounts (measured as concentration of dissolved organic carbon [DOC]) and simulated-sunlight UV-A intensities. Radical production rate was determined using fluorescence spectroscopy and was positively correlated with increases in TiO2 concentration and UV-A intensity, and negatively correlated with increased DOC concentration. Daphnia magna toxicity was also found to decrease with NOM addition, which is attributed to the decreased radical production rate with increased DOC concentrations. We demonstrate that the rate of ROS production from simulated-sunlight-irradiated TiO2 NPs can be quantified using relatively simple fluorescent techniques. We show that toxicity to TiO2 NP varies greatly with conditions, and that concentration alone is a poor predictor of toxicity. Describing toxicity/hydroxyl radical measurement may be a more accurate way to describe overall risk. We provide a framework for a simple model to describe toxicity/hydroxyl radical. These conclusions demonstrate the importance of considering exposure conditions as a means of risk management during TiO2 NP toxicity testing, waste management, and regulatory decisions. Environ Toxicol Chem 2021;40:1322-1334. © 2021 SETAC.