Patrick D. Guiney

Environmental release, environmental concentrations, and ecological risk of N,N-Diethyl-m-toluamide (DEET)

N,N-Diethyl-m-toluamides (DEET) commercial use as an insect repellent and other reported uses are reviewed. Evidence that DEET is reaching the environment mainly from consumer use of DEET-containing insect repellent includes studies reporting higher concentrations of DEET in surface water and wastewater samples during the summer months, the presence of DEET in on-site septic tank effluent at concentrations similar to that reported in wastewater treatment plant (WWTP) influent, and changes in WWTP effluent concentrations before and after the introduction of a DEET replacement in Germany. Its detected concentrations in influent and effluent of WWTP and surface water worldwide are reviewed and correlations between DEET usage and wastewater effluent concentrations are analyzed. The removability during wastewater treatment is also evaluated. A correlation between commercial DEET use in a metropolitan area and concentrations in WWTP effluents was assessed, and 2 different models were used to predict DEET concentrations in rivers and streams throughout the United States. Ecological toxicity data are reviewed for acute studies and for chronic values that are available for Daphnia magna and algae. The ecological risk of DEET usage is evaluated by examining the relationship of the expected dose/response to observed concentrations.

The Noninvasive Mouse Ear Swelling Assay II. Testing the Contact Sensitizing Potency of Fragrances

The noninvasive mouse ear swelling assay (MESA) for contact allergy testing was evaluated using fragrance components and complex fragrance mixtures. The test materials represented weak sensitizers and nonsensitizers. Two versions of the MESA were investigated. Both were noninvasive and utilized only topical abdominal dosing and ear challenge with single applications in BALB/cBy mice. The vit A MESA differed from the regular MESA only in that mice were maintained on a diet with 17-fold higher levels of vitamin A (vit A) acetate beginning 3 weeks prior to induction. Sensitization reactions were determined by measuring the mean increase in ear swelling over baseline at 24, 48 and 72 hr postexposure. Irritation dose-response curves facilitated choosing a high nonirritating challenge dose. Sensitization dose-response curves were developed for cinnamaldehyde (CINN) and a complex fragrance mixture, F-16. From these curves, the SD50 was determined. This value represents the dose which sensitized half the animals and serves to rank the potency of compounds for allergic contact dermatitis and to compare values among different assays. The SD50 for CINN was 21.6% while the SD50vit A for F-16 was 26.6%. The other fragrance, isoeugenol (ISOE), and fragrance mixtures, F-07 and F-22, were also found to be weak sensitizers in the MESA and vit A MESA. The results in the MESA for CINN and ISOE were in the range observed with guinea pig test protocols but showed that the MESA was more sensitive than human test protocols. Two of the fragrance mixtures tested in the MESA gave comparable results in the Buehler guinea pig assay. However, the third (F-22) was negative in the Buehler assay and the MESA, but positive in the vit A MESA. The results of this work with weak sensitizers and the companion study (Thorne et al., 1991) with potent sensitizers at low doses illustrate that the noninvasive MESA is as sensitive as many standard guinea pig assays. In addition, it is easier and much less expensive to perform. The vit A MESA has the sensitivity and predictive power needed to test compounds and mixtures for contact sensitizing potency.

Relevance Weighting of Tier 1 Endocrine Screening Endpoints by Rank Order

Weight of evidence (WoE) approaches are recommended for interpreting various toxicological data, but few systematic and transparent procedures exist. A hypothesis-based WoE framework was recently published focusing on the U.S. EPAs Tier 1 Endocrine Screening Battery (ESB) as an example. The framework recommends weighting each experimental endpoint according to its relevance for deciding eight hypotheses addressed by the ESB. Here we present detailed rationale for weighting the ESB endpoints according to three rank ordered categories and an interpretive process for using the rankings to reach WoE determinations. Rank 1 was assigned to in vivo endpoints that characterize the fundamental physiological actions for androgen, estrogen, and thyroid activities. Rank 1 endpoints are specific and sensitive for the hypothesis, interpretable without ancillary data, and rarely confounded by artifacts or nonspecific activity. Rank 2 endpoints are specific and interpretable for the hypothesis but less informative than Rank 1, often due to oversensitivity, inclusion of narrowly context-dependent components of the hormonal system (e.g., in vitro endpoints), or confounding by nonspecific activity. Rank 3 endpoints are relevant for the hypothesis but only corroborative of Ranks 1 and 2 endpoints. Rank 3 includes many apical in vivo endpoints that can be affected by systemic toxicity and nonhormonal activity. Although these relevance weight rankings (WREL ) necessarily involve professional judgment, their a priori derivation enhances transparency and renders WoE determinations amenable to methodological scrutiny according to basic scientific premises, characteristics that cannot be assured by processes in which the rationale for decisions is provided post hoc.

Environmental fate of pyrethroids in urban and suburban stream sediments and the appropriateness of Hyalella azteca model in determining ecological risk

According to several recent studies using standard acute Hyalella azteca sediment bioassays, increased pyrethroid use in urban and suburban regions in California has resulted in the accumulation of toxic concentrations of pyrethroids in sediments of area streams and estuaries. However, a critical review of the literature indicates that this is likely an overestimation of environmental risk. Hyalella azteca is consistently the most susceptible organism to both aqueous and sediment-associated pyrethroid exposures when compared to a suite of other aquatic taxa. In some cases, H. azteca LC50 values are less than the community HC10 values, suggesting that the amphipod is an overly conservative model for community- or ecosystem-level impacts of sediment-associated pyrethroids. Further, as a model for responses of field populations of H. azteca, the laboratory bioassays considerably overestimate exposure, because the amphipod is more appropriately characterized as an epibenthic organism, not a true sediment dweller; H. azteca preferentially inhabit aquatic macrophytes, periphyton mats, and leaf litter, which drastically reduces their exposure to contaminated sediments. Sediment-bound pyrethroids are transported via downstream washing of fine particulates resulting in longer range transport but also more efficient sequestration of the chemical. In addition, site-specific variables such as sediment organic carbon content, grain size, temperature, and microbial activity alter pyrethroid bioavailability, degradation, and toxicity on a microhabitat scale. The type and source of the carbon in particular, influences the pyrethroid sequestering ability of sediments. The resulting irregular distribution of pyrethroids in stream sediments suggests that sufficient nonimpacted habitat may exist as refugia for resident sediment-dwelling organisms for rapid recolonization to occur. Given these factors, we argue that the amphipod model provides, at best, a screening level assessment of pyrethroid impacts and can correctly identify those sediments not toxic to benthic organisms but cannot accurately predict where sediments will be toxic.

Assessment of an approach to estimating aquatic bioconcentration factors using reduced sampling

For a broad range of circumstances, we show that reliable bioconcentration factor (BCF) estimates can be made using a study design that is based on standard regulatory guideline test procedures but that uses significantly fewer animals and resources. This minimized design involves taking tissue samples only twice during a 14-d depuration period. The utility of the minimized test design was first assessed by resampling data from a series of standard guideline tests and calculating the BCF estimates that would have been obtained if the test had been performed using the minimized design. Data from 25 bioconcentration curves giving BCF estimates ranging from approximately 0.3 to over 20,000 were used. The correlation of log BCF estimates from the guideline study with log BCF estimates from the simulated minimized tests was r=0.99, and the slope of the regression line was 0.96. The robustness of BCF estimates to random variation in measurement of chemicals in fish and water (coefficients of variation of concentrations ranging up to 25%) was evaluated using Monte Carlo simulations. For chemicals with depuration half-lives of less than the length of the depuration period, median BCF estimates from the Monte Carlo simulations of the minimized design were always within 7% of the true BCE The ratio of the 95th to the 5th percentile BCF estimates was always less than or equal to 3.7. Furthermore, the span from the 95th to the 5th percentile of BCF estimates was only 15% wider in the minimized test than in the full guideline test, even though animal use and analytical effort was markedly reduced.

Recommended approaches to the scientific evaluation of ecotoxicological hazards and risks of endocrine-active substances

A SETAC Pellston Workshop® “Environmental Hazard and Risk Assessment Approaches for Endocrine-Active Substances (EHRA)” was held in February 2016 in Pensacola, Florida, USA. The primary objective of the workshop was to provide advice, based on current scientific understanding, to regulators and policy makers; the aim being to make considered, informed decisions on whether to select an ecotoxicological hazard- or a risk-based approach for regulating a given endocrinedisrupting substance (EDS) under review. The workshop additionally considered recent developments in the identification of EDS. Case studies were undertaken on 6 endocrine-active substances (EAS—not necessarily proven EDS, but substances known to interact directly with the endocrine system) that are representative of a range of perturbations of the endocrine system and considered to be data rich in relevant information at multiple biological levels of organization for 1 or more ecologically relevant taxa. The substances selected were 17α-ethinylestradiol, perchlorate, propiconazole, 17β-trenbolone, tributyltin, and vinclozolin. The 6 case studies were not comprehensive safety evaluations but provided foundations for clarifying key issues and procedures that should be considered when assessing the ecotoxicological hazards and risks of EAS and EDS. The workshop also highlighted areas of scientific uncertainty, and made specific recommendations for research and methods-development to resolve some of the identified issues. The present paper provides broad guidance for scientists in regulatory authorities, industry, and academia on issues likely to arise during the ecotoxicological hazard and risk assessment of EAS and EDS. The primary conclusion of this paper, and of the SETAC Pellston Workshop on which it is based, is that if data on environmental exposure, effects on sensitive species and life-stages, delayed effects, and effects at low concentrations are robust, initiating environmental risk assessment of EDS is scientifically sound and sufficiently reliable and protective of the environment. In the absence of such data, assessment on the basis of hazard is scientifically justified until such time as relevant new information is available.

Current limitations and recommendations to improve testing for the environmental assessment of endocrine active substances

In the present study, existing regulatory frameworks and test systems for assessing potential endocrine active chemicals are described, and associated challenges are discussed, along with proposed approaches to address these challenges. Regulatory frameworks vary somewhat across geographies, but all basically evaluate whether a chemical possesses endocrine activity and whether this activity can result in adverse outcomes either to humans or to the environment. Current test systems include in silico, in vitro, and in vivo techniques focused on detecting potential endocrine activity, and in vivo tests that collect apical data to detect possible adverse effects. These test systems are currently designed to robustly assess endocrine activity and/or adverse effects in the estrogen, androgen, and thyroid hormone signaling pathways; however, there are some limitations of current test systems for evaluating endocrine hazard and risk. These limitations include a lack of certainty regarding: 1) adequately sensitive species and life stages; 2) mechanistic endpoints that are diagnostic for endocrine pathways of concern; and 3) the linkage between mechanistic responses and apical, adverse outcomes. Furthermore, some existing test methods are resource intensive with regard to time, cost, and use of animals. However, based on recent experiences, there are opportunities to improve approaches to and guidance for existing test methods and to reduce uncertainty. For example, in vitro high-throughput screening could be used to prioritize chemicals for testing and provide insights as to the most appropriate assays for characterizing hazard and risk. Other recommendations include adding endpoints for elucidating connections between mechanistic effects and adverse outcomes, identifying potentially sensitive taxa for which test methods currently do not exist, and addressing key endocrine pathways of possible concern in addition to those associated with estrogen, androgen, and thyroid signaling. Integr Environ Assess Manag 2017;13:302-316.

A Comparative Human Health Risk Assessment of p-Dichlorobenzene-Based Toilet Rimblock Products Versus Fragrance/Surfactant-Based Alternatives

A comparison of the human health risk to consumers using one of two types of toilet rimblock products, either a p-dichlorobenzene-based rimblock or two newer fragrance/surfactant-based alternatives, was conducted. Rimblock products are designed for global use by consumers worldwide and function by releasing volatile compounds into indoor air with subsequent exposure presumed to be mainly by inhalation of indoor air. Using the THERdbASE exposure model and experimentally determined emission data, indoor air concentrations and daily intake values were determined for both types of rimblock products. Modeled exposure concentrations from a representative p-dichlorobenzene rimblock product are an order of magnitude higher than those from the alternative rimblock products due to its nearly pure composition and high sublimation rate. Lifetime exposure to p-dichlorobenzene or the subset of fragrance components with available RfD values is not expected to lead to non-cancer-based adverse health effects based on the exposure concentrations estimated using the THERdbASE model. A similar comparison of cancer-based effects was not possible as insufficient data were available for the fragrance components.

Inhibition of germinal vesicle breakdown in Xenopus oocytes in vitro by a series of substituted glycol ethers: GLYCOL ETHER INHIBITION OF OOCYTE MATURATION

A 24 hour in vitro Xenopus oocyte maturation (germinal vesicle breakdown [GVBD]) assay developed by Pickford and Morris (Environmental Health Perspectives, 1999, 107, 285–292) was used to screen a series of substituted glycol ethers (GEs). Substituted GEs included: ethylene glycol monomethyl ether (EGME); EG monoethyl ether (EGEE); EG monopropyl ether (EGPE); EG monobutyl ether (EGBE); EG monohexyl ether (EGHE); diethylene glycol monomethyl ether (DGME); triethylene glycol monomethyl ether (TGME); ethylene glycol monophenyl ether (EGPhE); EG monobenzyl ether (EGBeE); EG diphenyl ether (EGDPhE); and propylene glycol monophenyl ether (PGPhE). The GEs inhibited progesterone‐ or androstenedione‐induced GVBD with the following relative potency: EGPhE > PGPhE > EGME >> EGEE ≥ EGBeE > EGPE >> EGBE >EGHE > EGDPhE >> DGME ≥ TGME, or EGPhE >> PGPhE >> EGBeE > EGDPhE > EGEE > EGME > EGPE > EGBE, EGHE, DGME and TGME, respectively. Further, [3H]progesterone or [3H]androstenedione binding affinities to the oocyte plasma membrane progesterone receptor (OMPR) or classical androgen receptor (AR) were: EGME > EGPhE ≥ PGPhE ≥ EGEE > EGBeE >> EGPE >> EGBE ≥ EGHE > EGDPhE, TGME, and DGME, or EGPhE > PGPhE >> EGBeE > EGDPhE >> EGEE ≥ EGME >> EGPE, EGBE, and EGHE > DGME and TGME, respectively. Binary joint mixture studies with the GVBD model using flutamide (AR antagonist) and EGPhE indicated that flutamide/EGPhE mixture acted in a concentration additive manner. The effects of substituted GE series, however, may be mediated through the OMPR; the potency of EGPhE may be the result of bimodal inhibition of both the OMPR and AR pathways.

Evaluation of the developmental toxicity of perfluorooctanesulfonate in the Anuran, Silurana tropicalis : PFOS Induces Abnormal Development in West African Clawed Frog

A 150‐day post‐metamorphosis (dpm) partial lifecycle study exposing Silurana tropicalis to <0.03 (control), 0.06, 0.13 0.25, 0.5 and 1.0 mg/L perfluorooctanesulfonate (PFOS) was conducted. A subset of specimens from the control and each treatment were evaluated at metamorphic completion. A significant increase in the median metamorphosis time was observed in the 1.0 mg/L PFOS treatment relative to the control. A modest increase in the occurrence, but not severity, of mild follicular hypertrophy was found in thyroid glands from organisms exposed to the 0.62 and 1.1 mg/L PFOS treatments. At 150 dpm, a concentration‐dependent increase in whole body PFOS residues was measured ranging from 29.6 to 163.5 mg/kg in the 0.05 and 1.1 mg/L PFOS treatments. Decreased body weight and snout‐vent length were noted in specimens exposed to 1.1 mg PFOS/L at the completion of metamorphosis. Body weight was reduced in the 1.1 mg/L PFOS concentration; however, snout‐vent length was not affected by PFOS exposure at 150 dpm. An increased proportion of phenotypic males were noted in the 0.62 and 1.1 mg/L PFOS treatments. Abnormal ovary development characterized by size asymmetry, necrosis and formation of excessive fibrous connective tissue was identified in females exposed to 0.29 and 1.1 mg PFOS/L. Asymmetrically misshaped testes were found at 1.1 mg/L PFOS. Results suggested that PFOS is capable of interfering with S. tropicalis growth before metamorphic completion and growth and gonad development during juvenile development.