AOP Report: Adverse Outcome Pathways for Aromatase Inhibition or Androgen Receptor Agonism Leading to Male-Biased Sex Ratio and Population Decline in Fish.

Screening and testing of potential endocrine-disrupting chemicals for ecological effects are examples of risk assessment/regulatory activities that can employ adverse outcome pathways (AOPs) to establish linkages between readily measured alterations in endocrine function and whole organism- and population-level responses. Of particular concern are processes controlled by the hypothalamic-pituitary-gonadal/thyroidal (HPG/T) axes. However, the availability of AOPs suitable to meet this need is currently limited in terms of species and life-stage representation relative to the diversity of endpoints influenced by HPG/T function. In our report we describe two novel AOPs that comprise a simple AOP network focused on the effects of chemicals on sex differentiation during early development in fish. The first AOP (346) documents events starting with inhibition of cytochrome P450 aromatase (CYP19), resulting in decreased availability of 17ß-estradiol during gonad differentiation, which increases the occurrence of testis formation, resulting in a male-biased sex ratio and consequent population-level declines. The second AOP (376) is initiated by activation of the androgen receptor (AR), also during sexual differentiation, again resulting in a male-biased sex ratio and population-level effects. Both AOPs are strongly supported by existing physiological and toxicological evidence, including numerous fish studies with model CYP19 inhibitors and AR agonists. Accordingly, AOPs 346 and 376 provide a basis for more focused screening and testing of chemicals with the potential to affect HPG function in fish during early development. Environ Toxicol Chem 2023;42:747-756. Published 2023. This article is a U.S. Government work and is in the public domain in the USA.

Verification of In Vivo Estrogenic Activity for Four Per- and Polyfluoroalkyl Substances (PFAS) Identified as Estrogen Receptor Agonists via New Approach Methodologies.

Given concerns about potential toxicological hazards of the thousands of data-poor per- and polyfluorinated alkyl substances (PFAS) currently in commerce and detected in the environment, tiered testing strategies that employ high-throughput in vitro screening as an initial testing tier have been implemented. The present study evaluated the effectiveness of previous in vitro screening for identifying PFAS capable, or incapable, of inducing estrogenic responses in fish exposed in vivo. Fathead minnows (Pimephales promelas) were exposed for 96 h to five PFAS (perfluorooctanoic acid [PFOA]; 1H,1H,8H,8H-perfluorooctane-1,8-diol [FC8-diol]; 1H,1H,10H,10H-perfluorodecane-1,10-diol [FC10-diol]; 1H,1H,8H,8H-perfluoro-3,6-dioxaoctane-1,8-diol [FC8-DOD]; and perfluoro-2-methyl-3-oxahexanoic acid [HFPO-DA]) that showed varying levels of in vitro estrogenic potency. In agreement with in vitro screening results, exposure to FC8-diol, FC10-diol, and FC8-DOD caused concentration-dependent increases in the expression of transcript coding for vitellogenin and estrogen receptor alpha and reduced expression of insulin-like growth factor and apolipoprotein eb. Once differences in bioconcentration were accounted for, the rank order of potency in vivo matched that determined in vitro. These results provide a screening level benchmark for worst-case estimates of potential estrogenic hazards of PFAS and a basis for identifying structurally similar PFAS to scrutinize for putative estrogenic activity.

A Multidimensional Matrix Model for Predicting the Effects of Male-Biased Sex Ratios on Fish Populations.

Laboratory experiments have established that exposure to certain endocrine-active substances prior to and/or during the period of sexual differentiation can lead to skewed sex ratios in fish. However, the potential long-term population impact of biased sex ratio depends on multiple factors including the life history of the species and whether the ratio is male or female-biased. In the present study, we describe a novel multidimensional, density-dependent matrix model that analyzes age class-structure of both males and females over time, allowing for the quantitative evaluation of the effects of biased sex ratio on population status. This approach can be used in conjunction with field monitoring efforts and/or laboratory testing to link effects on sex ratio due to chemical and/or nonchemical stressors to adverse outcomes in whole organisms and populations. For demonstration purposes, we applied the model to evaluate population trajectories for fathead minnow (Pimephales promelas) exposed to prochloraz, an aromatase inhibitor, during sexual differentiation. The model also was used to explore the population impact in a more realistic exposure scenario in which both adult and early life stages of fish are exposed concurrently to prochloraz, which, in addition to altering sex ratio during development, can decrease vitellogenin and egg production in adult females. For each exposure scenario, the model was used to analyze total population size, numbers of females and of males, and sex specific recruitment of the F1 generation. The present study illustrates the utility of multidimensional matrix population models for ecological risk assessment in terms of integrating effects across a population of an organism even when chemical effects on individuals are manifested via different pathways depending on life stage. Environ Toxicol Chem 2022;41:1066-1077. Published 2022. This article is a U.S. Government work and is in the public domain in the USA.