What Approaches Should be Used to Prioritize Pharmaceuticals and Personal Care Products for Research on Environmental and Human Health Exposure and Effects?

Pharmaceuticals and personal care products (PPCPs) are released from multiple anthropogenic sources and thus have a ubiquitous presence in the environment. The environmental exposure and potential effects of PPCPs on biota and humans has aroused concern within the scientific community and the public. Risk assessments are commonly conducted to evaluate the likelihood of chemicals including PPCPs that pose health threats to organisms inhabiting various environmental compartments and humans. Because thousands of PPCPs are currently used, it is impractical to assess the environmental risk of all of them due to data limitations; in addition, new PPCPs are continually being produced. Prioritization approaches, based either on exposure, hazard, or risk, provide a possible means by which those PPCPs that are likely to pose the greatest risk to the environment are identified, thereby enabling more effective allocation of resources in environmental monitoring programs in specific geographical locations and ecotoxicological investigations. In the present review, the importance and current knowledge concerning PPCP occurrence and risk are discussed and priorities for future research are proposed, in terms of PPCP exposure (e.g., optimization of exposure modeling in freshwater ecosystems and more monitoring of PPCPs in the marine environment) or hazard (e.g., differential risk of PPCPs to lower vs. higher trophic level species and risks to human health). Recommended research questions for the next 10 years are also provided, which can be answered by future studies on prioritization of PPCPs. Environ Toxicol Chem 2022;00:1-14. © 2022 SETAC.

Comparison of false-positive rates of 2 hypothesis-test approaches in relation to laboratory toxicity test performance.

We compared 2 statistical hypothesis-test approaches (no-observed-effect concentration [NOEC] and test of significant toxicity [TST]) to determine the influence of laboratory test performance on the false-positive error rate using the US Environmental Protection Agency's Ceriodaphnia dubia reproduction whole-effluent toxicity (WET) test endpoint. Simulation and power calculations were used to determine error rates based on observed control coefficients of variation (CV) for 8 laboratories over a range of effect levels. Average C. dubia control reproduction among laboratories was 20 to 40 offspring per female, and the 75th percentile CV was 0.10 to 0.31, reflecting a range in laboratory performance. The 2 approaches behave similarly for CVs of 0.2 to 0.3. At effects <10%, as CV decreases, TST is less likely to declare toxicity and NOEC is more likely to do so. Laboratory performance affects whether a sample is declared toxic and influences the probability of false-positive (and -negative) error rates using either approach. At the 75th percentile control CV observed for each laboratory, 4 laboratories would achieve approximately a 5% false-positive rate using 13 or fewer replicates for this test method. For the remaining 4 laboratories, more replicates would be needed to achieve a 5% false-positive rate. The present analyses demonstrate how false-positive rates are influenced by laboratory performance and WET test design. Environ Toxicol Chem 2019;38:511-523. Published 2019 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

A framework for screening sites at risk from contaminants of emerging concern.

Trace levels of a variety of currently unregulated organic chemicals have been detected in treated wastewater effluents and surface waters that receive treated effluents. Many of these chemicals of emerging concern (CECs) originate from pharmaceuticals and personal care products that are used widely and that frequently are transported "down the drain" to a wastewater treatment plant (WWTP). Actual effects of CECs on aquatic life have been difficult to document, although biological effects consistent with effects of some CECs have been noted. There is a critical need to find appropriate ways to screen wastewater sites that have the greatest potential of CEC risk to biota. Building on the work of several researchers, the authors present a screening framework, as well as examples based on the framework, designed to identify high-risk versus lower-risk sites that are influenced by WWTP effluent. It is hoped that this framework can help researchers, utilities, and the larger water resource community focus efforts toward improving CEC risk determinations and management of these risks.

Predicted no effect concentration derivation as a significant source of variability in environmental hazard assessments of chemicals in aquatic systems: an international analysis.

Environmental hazard assessments for chemicals are carried out to define an environmentally "safe" level at which, theoretically, the chemical will not negatively affect any exposed biota. Despite this common goal, the methodologies in use are very diverse across different countries and jurisdictions. This becomes particularly obvious when international scientists work together on documents with global scope, e.g., in the World Health Organization (WHO) International Program on Chemical Safety. In this article, we present a study that describes the extent of such variability and analyze the reasons that lead to different outcomes in deriving a "safe level" (termed the predicted no effect concentration [PNEC] throughout this article). For this purpose, we chose 5 chemicals to represent well-known substances for which sufficient high-quality aquatic effects data were available: ethylene glycol, trichloroethylene, nonylphenol, hexachlorobenzene, and copper (Cu). From these data, 2 data sets for each chemical were compiled: the full data set, that contained all information from selected peer-review sources, and the base data set, a subsample of the full set simulating limited data. Scientists from the European Union (EU), United States, Canada, Japan, and Australia independently carried out hazard assessments for each of these chemicals using the same data sets. Their reasoning for key study selection, use of assessment factors, or use of probabilistic methods was comprehensively documented. The observed variation in the PNECs for all chemicals was up to 3 orders of magnitude, and this was not simply due to obvious factors such as the size of the data set or the methodology used. Rather, this was due to individual decisions of the assessors within the scope of the methodology used, especially key study selection, acute versus chronic definitions, and size of assessment factors. Awareness of these factors, together with transparency of the decision-making process, would be necessary to minimize confusion and uncertainty related to different hazard assessment outcomes, particularly in international documents. The development of a "guideline on transparency in decision-making" ensuring the decision-making process is science-based, understandable, and transparent, may therefore be a promising way forward.

Evaluation of the test of significant toxicity for determining the toxicity of effluents and ambient water samples.

The test of significant toxicity (TST) is a hypothesis-testing approach based on bioequivalence developed by the U.S. Environmental Protection Agency (U.S. EPA) for analyzing whole-effluent toxicity (WET) and ambient toxicity data. The present study compares results of acute and chronic toxicity tests of effluent, storm-water, and ambient (i.e., receiving-water) samples using both the TST and the standard no-observed-effect concentration (NOEC) approach. Valid WET data were analyzed from 890 tests provided by more than 25 dischargers in California and Washington, USA, representing the majority of test methods used in the U.S. WET program. An additional 3,201 freshwater chronic toxicity tests, obtained from ambient monitoring programs in California, were also analyzed. The TST and NOEC approaches both declared a low number (<6.5%) of tests toxic if effects were below the unacceptable toxicity regulatory management decision (RMD) of 25% effect in chronic tests or 20% effect in acute tests. However, those test methods having generally lower within-test variability and greater test power (e.g., urchin fertilization test) had a much lower percentage of tests declared toxic than the NOEC approach when effects were below the unacceptable toxicity RMD. In addition, the TST showed fewer tests to be nontoxic than NOEC if the test exhibited effects greater than the toxicity RMD (0.1 and 9.6% for TST and NOEC, respectively, for effluents and 0 and 9.5%, respectively, for ambient samples). Our results demonstrate that the TST is more likely to identify a toxic sample when effects are fairly substantial (≥ 25% effect in chronic testing and ≥ 20% effect in acute tests) and less likely to identify a sample as toxic when effects are negligible (≤ 10% effect). Furthermore, these results demonstrate that appropriate WET data interpretation benefits from having well-designed test methods with sufficient power to identify significant toxicity or biologically insignificant effects when they occur.

Evaluation of whole effluent toxicity data characteristics and use of Welch's T-test in the test of significant toxicity analysis.

The U.S. Environmental Protection Agency (U.S. EPA) and state agencies evaluate the toxicity of effluent and surface water samples based on statistical endpoints derived from multiconcentration tests (e.g., no observed effect concentration, EC25). The test of significant toxicity (TST) analysis is a two-sample comparison test that uses Welch's t test to compare organism responses in a sample (effluent or surface water) with responses in a control or site sample. In general, any form of t test (Welch's t included) is appropriate only if the data meet assumptions of normality and homogeneous variances. Otherwise, nonparametric tests are recommended. TST was designed to use Welch's t as the statistical test for all whole effluent toxicity (WET) test data. The authors evaluated the suitability of using Welch's t test for analyzing two-sample toxicity (WET) data, and within the TST approach, by examining the distribution and variances of data from over 2,000 WET tests and by conducting multiple simulations of WET test data. Simulated data were generated having variances and nonnormal distributions similar to observed WET test data for control and the effluent treatment groups. The authors demonstrate that (1) moderately unequal variances (similar to WET data) have little effect on coverage of the t test or Welch t test (for normally distributed data), and (2) for nonnormally distributed data (similar in distribution to WET data) TST, using Welch's t test, has close to nominal coverage on the basis of simulations with up to a ninefold difference in variance between the effluent and control groups (∼95th percentile based on observed WET test data).

It is time for changes in the analysis of whole effluent toxicity data.

The whole effluent toxicity (WET) program in the United States, Canada, and other countries typically requires multi concentration testing of effluents. While multiconcentration testing of chemicals is desirable for regulatory and scientific reasons, we believe this requirement is not as efficient for evaluating effluent compliance in a WET program. The key regulatory question of concern is whether an effluent is toxic or not, which is best answered statistically using a hypothesis approach, not a point estimate approach. However, the traditional hypothesis approach currently recommended does not reward high within-test precision. This report describes the need for 3 specific changes in the analysis of WET compliance data that we believe would yield a more robust WET regulatory program: (1) restate the null hypothesis so that test power is associated with demonstrating that the effluent is not toxic, (2) use USEPA's Test of Significant Toxicity (based on the noninferiority approach) to identify unacceptable toxicity as well as acceptable effects with a high probability, and (3) evaluate only the test control and the critical concentration of concern (e.g., instream waste concentration). We demonstrate that instituting these 3 changes would provide: Positive incentives for permittees to produce high-quality WET data, a transparent analysis approach in which the permittee could have greater control over regulatory decisions based on test results, and potentially a less expensive testing program because fewer effluent concentrations need to be examined within a test. As a result, WET test frequency could be increased for the same cost as current testing programs while providing greater representativeness of effluent quality.

An approach for determining bioassessment performance and comparability.

Many organizations in the USA collect aquatic bioassessment data using different sampling and analysis methods, most of which have unknown performance in terms of data quality produced. Thus, the comparability of bioassessments produced by different organizations is often unknown, ultimately affecting our ability to make comprehensive assessments on large spatial scales. We evaluated a pilot approach for determining bioassessment performance using macroinvertebrate data obtained from several states in the Southeastern USA. Performance measures evaluated included precision, sensitivity, and responsiveness to a human disturbance gradient, defined in terms of a land disturbance index value for each site, combined with a value for specific conductance, and instream habitat quality. A key finding of this study is the need to harmonize ecoregional reference conditions among states so as to yield more comparable and consistent bioassessment results. Our approach was also capable of identifying potential areas for refinement such as reevaluation of less precise, sensitive, or responsive metrics that may result in suboptimal index performance. Higher performing bioassessments can yield information beyond "impaired" versus "unimpaired" condition. Acknowledging the limitations of this pilot study, we would recommend that performance evaluations use at least 50 sites, 10 of which are ecoregional reference sites. Efforts should be made to obtain data from the entire human disturbance gradient in an ecoregion to improve statistical confidence in performance measures. Having too few sites will result in an under-representation of certain parts of the disturbance gradient (e.g., too few poor quality sites), which may bias sensitivity and responsiveness estimates.

Test of significant toxicity: a statistical application for assessing whether an effluent or site water is truly toxic.

The U.S. Environmental Protection Agency (U.S. EPA) and state agencies implement the Clean Water Act, in part, by evaluating the toxicity of effluent and surface water samples. A common goal for both regulatory authorities and permittees is confidence in an individual test result (e.g., no-observed-effect concentration [NOEC], pass/fail, 25% effective concentration [EC25]), which is used to make regulatory decisions, such as reasonable potential determinations, permit compliance, and watershed assessments. This paper discusses an additional statistical approach (test of significant toxicity [TST]), based on bioequivalence hypothesis testing, or, more appropriately, test of noninferiority, which examines whether there is a nontoxic effect at a single concentration of concern compared with a control. Unlike the traditional hypothesis testing approach in whole effluent toxicity (WET) testing, TST is designed to incorporate explicitly both α and ß error rates at levels of toxicity that are unacceptable and acceptable, given routine laboratory test performance for a given test method. Regulatory management decisions are used to identify unacceptable toxicity levels for acute and chronic tests, and the null hypothesis is constructed such that test power is associated with the ability to declare correctly a truly nontoxic sample as acceptable. This approach provides a positive incentive to generate high-quality WET data to make informed decisions regarding regulatory decisions. This paper illustrates how α and ß error rates were established for specific test method designs and tests the TST approach using both simulation analyses and actual WET data. In general, those WET test endpoints having higher routine (e.g., 50th percentile) within-test control variation, on average, have higher method-specific α values (type I error rate), to maintain a desired type II error rate. This paper delineates the technical underpinnings of this approach and demonstrates the benefits to both regulatory authorities and permitted entities.

Evaluation of effluent toxicity as an indicator of aquatic life condition in effluent-dominated streams: a pilot study.

The types and quality of data needed to determine relationships between chronic whole effluent toxicity (WET) test results and in-stream biological condition were evaluated using information collected over a 1.5-y period from 6 different sites across the United States. A data-quality-objectives approach was used that included several proposed measurement quality objectives (MQOs) that specified desired precision, bias, and sensitivity of methods used. The 6 facilities used in this study (4 eastern and 2 western United States) all had design effluent concentrations >60% of the stream flow. In addition to at least quarterly chronic Ceriodaphnia dubia, Pimephales promelas (fathead minnow), and Selenastrum capricornutum (green algae) WET tests, other tests were conducted to address MQOs, including splits, duplicates, and blind positive and negative controls. Macroinvertebrate, fish, and periphyton bioassessments were conducted at multiple locations upstream and downstream of each facility. The test acceptance criteria of the US Environmental Protection Agency (USEPA) were met for most WET tests; however, this study demonstrated the need to incorporate other MQOs (minimum and maximum percent significant difference and performance on blind samples) to ensure accurate interpretation of effluent toxicity. More false positives, higher toxicity, and more "failed" (noncompliant) tests were observed using no-observed-effect concentration (NOEC) as compared to the IC25 endpoint (concentration causing > or =25% decrease in organism response compared to controls). Algae tests often indicated the most effluent toxicity in this study; however, this test was most susceptible to false positives and high interlaboratory variability. Overall, WET test results exhibited few relationships with bioassessment results even when accounting for actual effluent dilution. In general, neither frequency of WET noncompliance nor magnitude of toxicity in tests were significantly related to differences in biological condition upstream and downstream of a discharge. Periphyton assessments were most able to discriminate small changes downstream of the effluent, followed by macroinvertebrates and fish. Although sampling methods were robust, more replicate samples collected upstream and downstream of each facility were needed to increase detection power. In general, macroinvertebrate and periphyton assessments together appeared to be sufficient to address project objectives.

Toxicity models of pulsed copper exposure to Pimephales promelas and Daphnia magna.

Semiempirical models are useful for interpreting the response of aquatic organisms to toxicants as a function of exposure concentration and duration. Most applications predict cumulative mortality at the end of the test for constant exposure concentrations. Summary measures, such as the median lethal concentration, are then estimated as a function of concentration. Real-world exposures are not constant. Effects may depend on pulse timing, and cumulative analysis based only on integrated exposure concentration is not sufficient to interpret results. We undertook a series of pulsed-exposure experiments using standard toxicological protocols and interpreted the results (mortality, biomass, and reproduction) using a dynamic generalization of a Mancini/Breck--type model that includes two compartments, one for internal concentration as a function of exposure and one for site-of-action concentration or accumulated damage as a function of the internal dose. At exposure concentrations near the effects level, the model explained approximately 50% of the variability in the observed time history of survival, 43% of the change in biomass, and 83% of the variability in net reproduction. Unexplained variability may result from differences in organism susceptibility, amplified by the effects of small sample sizes in standard tests. The results suggest that response is sensitive to prior conditions and that constant-exposure experiments can underestimate the risk from intermittent exposures to the same concentration. For pulsed exposures, neither the average nor the maximum concentration alone is an adequate index of risk, which depends on both the magnitude, duration, and timing of exposure pulses. Better understanding about the impacts of pulsed exposures will require use of experimental protocols with significantly greater numbers of replicates.

Effects of pulsed copper exposures on early life-stage Pimephales promelas.

Effects of pulsed copper exposures were investigated using Pimephales promelas aged less than 24 h in short-term chronic testing (7 or 14 d) with moderately hard synthetic water. Concentrations tested were between the species mean chronic value (22 microg/L at a hardness of 100 mg/L as CaCO3) and the 7-d continuous exposure EC50 for survival (40 microg/L) to examine exposures that were not acutely toxic and representative of actual wastewater discharge permit exceedences. Factors tested included pulse duration, recovery time between pulses, and pulse frequency. Survival was the main endpoint affected in all treatments (analysis of variance, p < 0.05). Effects on fish biomass, independent of survival effects, were observed in only 2 of 86 treatments examined. Fish survival was negatively affected at average copper concentrations between 7 and 50% of the 7-d continuous exposure EC50. Exposures having a 48- to 96-h recovery time between pulses had less effect on fish survival than did treatments with shorter (12-24 h) or longer (>120 h) recovery times. Results suggest that the criteria averaging periods used in the United States, and the averaging periods typically used in wastewater discharge permit limits for copper, may not protect against effects of certain pulsed exposures.