Development and Initial Testing of EcoToxChip, a Novel Toxicogenomics Tool for Environmental Management and Chemical Risk Assessment.

New approach methods (NAMs) are increasingly important to help accelerate the pace of ecological risk assessment and offer more ethical, affordable, and efficient alternatives to traditional toxicity tests. In the present study, we describe the development, technical characterization, and initial testing of a toxicogenomics tool, EcoToxChip (384-well quantitative polymerase chain reaction [qPCR] array), to support chemical management and environmental monitoring for three laboratory model species-fathead minnow (Pimephales promelas), African clawed frog (Xenopus laevis), and Japanese quail (Coturnix japonica). Chip design, including gene selection, was informed by a diverse end-user group and quality control metrics (e.g., primer assay, reverse transcription, and PCR efficiency) performed well based on a priori established criteria. Correlation with RNA sequencing (seq) data provided additional confidence in this novel toxicogenomics tool. Although the present study represents an initial testing of only 24 EcoToxChips for each of the model species, the results provide increased confidence in the robustness/reproducibility of EcoToxChips for evaluating perturbations in gene expression associated with chemical exposure and thus, this NAM, combined with early-life stage toxicity testing, could augment current efforts for chemical prioritization and environmental management. Environ Toxicol Chem 2023;42:1763-1771. © 2023 SETAC.

Maternal Transfer and Apical and Physiological Effects of Dietary Hexabromocyclododecane Exposure in Parental Fathead Minnows (Pimephales promelas).

Hexabromocyclododecane (HBCD) is a persistent organic pollutant that has been characterized as an endocrine disruptor, undergoes maternal transfer, and hinders development and growth in oviparous organisms. The present study examined the apical effects of dietary HBCD (11.5, 36.4, 106 mg/kg, wet wt) on adult fathead minnow exposed for 49 days and the subsequent accumulation and maternal transfer kinetics in adult tissue and eggs, respectively. Exposed adults displayed a significant increase in egg production in the medium treatment group, but no other significant effects were noted. Maternal transfer of dietary HBCD had a similar egg-to-muscle ratios (EMR) in the low and medium treatment groups (1.65 and 1.27 [wet wt], respectively). However, the high treatment group deviated from other treatments with an EMR of 4.2 (wet wt), potentially due to differences in total lipid content in food and/or reaching diffusion/lipid saturation limits in adult tissue, resulting in lower accumulation in the adult muscle tissue. A positive correlation was observed between egg HBCD concentration and time of exposure, which indicates that maternal transfer of HBCD is of concern in fish, and further studies should be conducted to fully elucidate the potential adverse effects that may be observed in the early life stage of oviparous organisms. Environ Toxicol Chem 2023;42:143-153. © 2022 SETAC.

Tissue-specific selenium accumulation and toxicity in adult female Xenopus laevis chronically exposed to elevated dietary selenomethionine.

Selenium (Se) is a developmental toxicant that is also capable of altering the bioenergetic and endocrine status of adult fish. To date, aquatic ecotoxicological research has predominantly focused on the toxic effects of Se in fish, and minimal information has been published related to amphibians. The objective of the present study was to investigate the potential toxicity associated with chronically elevated dietary Se consumption in adult female amphibians utilizing the model species Xenopus laevis. Adult X. laevis females were fed a diet augmented with L-selenomethionine at measured concentrations of 0.7 µg Se/g (control), 10.9 µg Se/g, 30.4 µg Se/g, or 94.2 µg Se/g dry mass for 68 d, after which they were bred with untreated males. Ovary, egg, liver, muscle, and blood samples were collected from female frogs after completion of the exposure period and subsequent breeding to ascertain Se tissue distribution, muscle and liver triglyceride and glycogen levels, and plasma cortisol concentrations. The concentrations of Se measured in female tissues excluding the liver were significantly increased in proportion with dietary intake. No significant differences were observed among treatment groups with respect to biometric indices, energy stores, or stress response of adult female X. laevis after Se exposure, which suggests that this amphibian species is capable of accumulating substantial quantities of this element in their tissues with no adverse effects on fitness. Environ Toxicol Chem 2017;36:1047-1055. © 2016 SETAC.

Effects of Elevated In Ovo Selenium Exposure on Late Stage Development of Xenopus laevis Tadpoles.

The effects of elevated in ovo selenium (Se) exposure on late stage larval Xenopus laevis development were investigated. Adult females were fed diets augmented with selenomethionine for 68 days and bred with untreated males to obtain embryos with measured concentrations of 1.6 (control), 10.8, 28.1 and 81.7 µg Se/g dry mass. Embryos were reared under uncontaminated conditions until 50 % of individuals within an aquarium had completed metamorphosis. The highest in ovo Se exposure group exhibited greater froglet body mass and snout to vent length while having a higher proportion of tadpoles at earlier stages of development. No differences were detected among treatment groups for mortality or metamorphic timing during the rearing period. This research suggests that in ovo Se exposure has minimal effect on the survival and development of late stage larval X. laevis in a controlled laboratory environment with adequate food availability.

Dose-Dependent Early Life Stage Toxicities in Xenopus laevis Exposed In Ovo to Selenium.

Selenium (Se) is a developmental toxicant in oviparous vertebrates. The adverse reproductive effects of Se toxicity have been predominantly investigated in fishes and birds with only a few studies focusing on amphibians. The objective of this study was to determine tissue-based toxicity thresholds for early life stage Se toxicities in Xenopus laevis as a consequence of in ovo exposure through maternal transfer of dietary Se. Following a 68-day dietary exposure to food augmented with l-selenomethionine (SeMet) at measured concentrations of 0.7 (control), 10.9, 30.4, or 94.2 µg Se/g dry mass (d.m.), adult female X. laevis were bred with untreated males, and resulting embryos were incubated until 5 days postfertilization (dpf). The measured Se concentrations in eggs were 1.6, 10.8, 28.1, and 81.7 µg Se/g d.m., respectively. No biologically significant effects were observed on fertilization success, hatchability, or mortality in offspring. Frequency and severity of morphological abnormalities were significantly greater in 5 dpf tadpoles from the highest exposure group when compared to the control, with eye lens abnormalities being the most prominent of all abnormalities. The estimated EC10 value for frequency of total early life stage abnormalities was 44.9 µg Se/g egg d.m., which suggests that this amphibian species is less sensitive to in ovo Se exposure than most of the fish species studied to date.

Reduced swim performance and aerobic capacity in adult zebrafish exposed to waterborne selenite.

Although dietary exposure of adult fish to organoselenium in contaminated aquatic ecosystems has been reported to bioaccumulate and cause larval deformities in offspring, subtle physiological effects produced through low level waterborne selenium exposure in fish such as swim performance and aerobic capacity have not been investigated. To evaluate potential effects of selenite on these responses, adult zebrafish (Danio rerio) were exposed to nominal aqueous concentrations of 0, 10 or 100 µg/L sodium selenite for 14 days. Upon completion of the exposure period, fish underwent two successive swim trials in a swim tunnel respirometer to determine critical swim speed (Ucrit), oxygen consumption (MO2), standard and active metabolic rates, aerobic scope (AS) and cost of transport (COT) followed by analysis of whole body triglyceride and glycogen concentrations. Selenite exposure had a significant negative effect on Ucrit and aerobic capacity. Active metabolic rates and AS significantly decreased in both selenite exposure groups after the second swim trial. No significant effect was observed in MO2, standard metabolic rate, COT, triglyceride and glycogen levels, or condition factor between groups. These results suggest that aqueous selenite exposure at environmentally relevant concentrations produces adverse effects on aerobic capacity that can diminish endurance and maximum swim speeds, which may lower fish survivability.