Identifying transcriptomic indicators of tertiary treated municipal effluent in longnose dace (Rhinichthys cataractae) caged under semi-controlled conditions in experimental raceways.

To evaluate effects of tertiary treated wastewater treatment plant effluent (MWWE) on transcriptomic responses in longnose dace (Rhinichthys cataractae; LND) we conducted a semi-controlled study in experimental raceways (Advancing Canadian Water Assets facility) imbedded in the Pine Creek treatment plant (Calgary, AB). LND collected from a reference site in the Bow River (REF) were caged in raceways containing either 5 % Pine Creek effluent (PC) or Bow River water (BR; control) over 28 d. Liver transcriptomes were analyzed in males and females sampled on days 7, 14 and 28 from BR and PC, and compared to REF fish on day 0. Concurrent with the caging, selected environmental substances of concern were analyzed in the BR and PC. Significantly different unigenes (SDUs) in females (vs males) within both BR and PC raceways increased over time and compared to REF fish. Moreover, SDUs in females and males within the same treatment (i.e., BR, PC) showed a temporal increase as well as compared to REF fish. Time was the dominant factor affecting SDUs, whereas sex and treatment had less of an impact on the transcriptome profiling. Gene Set Enrichment Analysis of BR vs PC over time revealed effects on genes involved in growth, metabolism of carbohydrates and lipids, and immune system on day 7; however, by day 28, 80-100 % of the transcripts localized to enriched biomarkers were associated with tissue immune responses in both sexes. Exposure to 5 % effluent had significant effects on female liver somatic index but no effects were observed on other phenotypic health indices in either sex. BR was used as the source of reference water, but analyses showed trace amounts of ESOCs. Analyses did not point towards definitive response patterns that could be used in field-based ecotoxicogenomic studies on the impacts of well-treated MWWE but suggested compromised adaptive immune responses.

Examination of recent hydroelectric dam projects in Canada for alignment of baseline studies, predictive modeling, and postdevelopment monitoring phases of aquatic environmental impact assessments.

Environmental impact assessment (EIA) has been widely criticized by the aquatic science community for poorly aligned approaches when selecting endpoints and collecting data during the baseline, predictive modeling, and postdevelopment monitoring phases. If these critical phases of the EIA process are not aligned properly, it can be difficult to evaluate the presence of postdevelopment effects. Examples of the misalignment of these phases include baseline studies failing to measure indicators that are monitored postdevelopment; predictive assessments that do not quantitatively predict conditions or potential impacts postdevelopment; and the failure to identify relevant indicators that may detect effects postdevelopment. For aquatic assessments, understanding how to protect critical ecosystem attributes to satisfy regulatory concerns could help to better align aquatic science monitoring activities across EIA phases. In this article we investigate recent Canadian hydroelectric dam EIAs to evaluate how well recent assessment approaches are meeting these necessary conditions of good aquatic EIA practice through the lens of ecosystem services from a fish's perspective. We found that larger facilities generally had baseline studies and modeling that better supported postdevelopment monitoring, but improvements in structure, linkages, and expectations would better align EIA phases in a manner that would improve assessments and environmental protection. Integr Environ Assess Manag 2024;20:616-644. © 2023 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Combining chemical, bioanalytical and predictive tools to assess persistence, seasonality, and sporadic releases of organic micropollutants within the urban water cycle.

Using a novel liquid chromatography-tandem mass spectrometry method with large volume direct injection and quantitation via isotope dilution, we evaluated the presence of 55 organic micropollutants in wastewater effluents, and locations within the Bow River and Elbow River watersheds in and around the city of Calgary, Alberta, Canada. In addition to establishing baseline micropollutant data for water utility operations, our study aimed to enhance our understanding of micropollutant behavior in the urban water cycle, assess the contributions of three wastewater treatment plants (WWTPs) to downstream receiving waters, explain the potential causes of total estrogenicity measured using the yeast-estrogen screen assay (YES), and prioritize a subset of substances for continuous monitoring. With data spanning 48 months and 95 river km, our results indicate the extensive persistence of metformin (antidiabetic), seasonality of N,N­diethyl-m-toluamide (DEET, insect repellant), O-desmethylvenlafaxine (antidepressant metabolite), and sulfamethoxazole (antibiotic) in source waters, and sporadic detections of a well-known perfluoroalkyl substance (PFOA). The seasonality of pharmaceuticals at the sentinel downstream monitoring site appeared to coincide with river dilution while that of DEET was likely attributable to peak usage during the warmer months. Steroidal estrogens were rarely detected in wastewater effluents although total estrogenicity via YES was evident, suggesting the presence of less potent but more abundant non-steroidal estrogens (e.g., flame retardants, bisphenols, and phthalates). A conservative mass balance analysis suggests that the largest WWTP (serving a population of >1 million) consistently contributed the highest load of micropollutants, with the exception of metformin, which appeared to be influenced by a smaller WWTP (serving 115,000) that operates a different activated sludge process. We consider metformin, sucralose, diclofenac, and venlafaxine as more effective conservative tracers of wastewater pollution due to their notably higher concentrations and persistence in the Bow River compared to carbamazepine and caffeine, respectively. Finally, hierarchical clustering revealed a close association between E. coli and caffeine, supporting the use of caffeine as an indicator of short-term, untreated anthropogenic inputs. Overall, this study yields valuable insights on the presence, behavior, and sources of organic micropollutants in the urban water cycle and identifies indicators of anthropogenic impacts that are useful for prioritizing future monitoring campaigns in Calgary and elsewhere.

Considering Fish as Recipients of Ecosystem Services Provides a Framework to Formally Link Baseline, Development, and Post-operational Monitoring Programs and Improve Aquatic Impact Assessments for Large Scale Developments.

In most countries, major development projects must satisfy an Environmental Impact Assessment (EIA) process that considers positive and negative aspects to determine if it meets environmental standards and appropriately mitigates or offsets negative impacts on the values being considered. The benefits of before-after-control-impact monitoring designs have been widely known for more than 30 years, but most development assessments fail to effectively link pre- and post-development monitoring in a meaningful way. Fish are a common component of EIA evaluation for both socioeconomic and scientific reasons. The Ecosystem Services (ES) concept was developed to describe the ecosystem attributes that benefit humans, and it offers the opportunity to develop a framework for EIA that is centred around the needs of and benefits from fish. Focusing an environmental monitoring framework on the critical needs of fish could serve to better align risk, development, and monitoring assessment processes. We define the ES that fish provide in the context of two common ES frameworks. To allow for linkages between environmental assessment and the ES concept, we describe critical ecosystem functions from a fish perspective to highlight potential monitoring targets that relate to fish abundance, diversity, health, and habitat. Finally, we suggest how this framing of a monitoring process can be used to better align aquatic monitoring programs across pre-development, development, and post-operational monitoring programs.

A 30-Year Study of Impacts, Recovery, and Development of Critical Effect Sizes for Endocrine Disruption in White Sucker (Catostomus commersonii) Exposed to Bleached-Kraft Pulp Mill Effluent at Jackfish Bay, Ontario, Canada.

Jackfish Bay is an isolated bay on the north shore of Lake Superior, Canada that has received effluent from a large bleached-kraft pulp mill since the 1940s. Studies conducted in the late 1980s found evidence of reductions in sex steroid hormone levels in multiple fish species living in the Bay, and increased growth, condition and relative liver weights, with a reduction in internal fat storage, reduced gonadal sizes, delayed sexual maturation, and altered levels of circulating sex steroid hormones in white sucker (Catostomus commersonii). These early studies provided some of the first pieces of evidence of endocrine disruption in wild animals. Studies on white sucker have continued at Jackfish Bay, monitoring fish health after the installation of secondary waste treatment (1989), changes in the pulp bleaching process (1990s), during facility maintenance shutdowns and during a series of facility closures associated with changing ownership (2000s), and were carried through to 2019 resulting in a 30-year study of fish health impacts, endocrine disruption, chemical exposure, and ecosystem recovery. The objective of the present study was to summarize and understand more than 75 physiological, endocrine, chemical and whole organism endpoints that have been studied providing important context for the complexity of endocrine responses, species differences, and challenges with extrapolation. Differences in body size, liver size, gonad size and condition persist, although changes in liver and gonad indices are much smaller than in the early years. Population modeling of the initial reproductive alterations predicted a 30% reduction in the population size, however with improvements over the last couple of decades those population impacts improved considerably. Reflection on these 30 years of detailed studies, on environmental conditions, physiological, and whole organism endpoints, gives insight into the complexity of endocrine responses to environmental change and mitigation.

Endocrine Disruptor Impacts on Fish From Chile: The Influence of Wastewaters.

Industrial wastewaters and urban discharges contain complex mixtures of chemicals capable of impacting reproductive performance in freshwater fish, called endocrine-disrupting compounds (EDCs). In Chile, the issue was highlighted by our group beginning over 15 years ago, by analyzing the impacts of pulp and paper mill effluents (PPME) in the Biobio, Itata, and Cruces River basins. All of the rivers studied are important freshwater ecosystems located in the Mediterranean region of Central Chile, each with a unique fish biodiversity. Sequentially, we developed a strategy based on laboratory assays, semicontrolled-field experiments (e.g., caging) and wild fish population assessments to explore the issue of reproductive impacts on both introduced and native fish in Chile. The integration of watershed, field, and laboratory studies was effective at understanding the endocrine responses in Chilean freshwater systems. The studies demonstrated that regardless of the type of treatment, pulp mill effluents can contain compounds capable of impacting endocrine systems. Urban wastewater treatment plant effluents (WWTP) were also investigated using the same integrated strategy. Although not directly compared, PPME and WWTP effluent seem to cause similar estrogenic effects in fish after waterborne exposure, with differing intensities. This body of work underscores the urgent need for further studies on the basic biology of Chilean native fish species, and an improved understanding on reproductive development and variability across Chilean ecosystems. The lack of knowledge of the ontogeny of Chilean fish, especially maturation and sexual development, with an emphasis on associated habitats and landscapes, are impediment factors for their conservation and protection against the threat of EDCs. The assessment of effects on native species in the receiving environment is critical for supporting and designing protective regulations and remediation strategies, and for conserving the unique Chilean fish biodiversity.

Methylmercury biomagnification in coastal aquatic food webs from western Patagonia and western Antarctic Peninsula.

Mercury (Hg) is a global pollutant of concern because its organic and more toxic form, methylHg (MeHg), bioaccumulates and biomagnifies through aquatic food webs to levels that affect the health of fish and fish consumers, including humans. Although much is known about trophic transfer of MeHg in aquatic food webs at temperate latitudes in the northern hemisphere, it is unclear whether its fate is similar in biota from coastal zones of the southeastern Pacific. To assess this gap, MeHg, total Hg and food web structure (using δ13C and δ15N) were measured in marine macroinvertebrates, fishes, birds, and mammals from Patagonian fjords and the Antarctic Peninsula. Trophic magnification slopes (TMS; log MeHg versus δ15N) for coastal food webs of Patagonia were high when compared with studies in the northern hemisphere, and significantly higher near freshwater inputs as compared to offshore sites (0.244 vs 0.192). Similarly, in Antarctica, the site closer to glacial inputs had a significantly higher TMS than the one in the Southern Shetland Islands (0.132 vs 0.073). Composition of the food web also had an influence, as the TMS increased when mammals and seabirds were excluded (0.132-0.221) at a coastal site. This study found that both the composition of the food web and the proximity to freshwater outflows are key factors influencing the TMS for MeHg in Patagonian and Antarctic food webs.

Assessing reproductive effects on fish populations: an evaluation of methods to predict the reproductive strategy of fishes.

Many environmental monitoring programs include an assessment of the health of fish populations using a sentinel species and include an indicator of reproductive potential. Knowledge of the reproductive strategy of the fish species is critical for data interpretation but is not always known. The reproductive strategy of a species can be determined from detailed histological analyses of ovaries throughout the reproductive cycle; however, these studies can be costly and can delay the implementation of a monitoring program. Three quick and cost-effective methods of predicting the reproductive strategy (annual single spawning or annual multiple spawning) are evaluated in this study using predicted probabilities from binary logistic regression models as a means of classifying the reproductive strategies of 18 different fish species in Atlantic Canada. The first method was based on the hypothesis that the variability in the ovary weight-body weight relationship in prespawning females is higher in multiple spawners. This method did not have a good classification rate due to some multiple spawners having low variability. The other two methods involved predictor variables representing the proportion of oocytes in different stages of development and predictor variables representing the distribution of oocyte sizes during the prespawning season for 111 fish (25 different samples for species). Predicted probabilities from these regression models could be used to correctly classify the reproductive strategies of all 25 samples (development stage model) and all but one sample (oocyte size distribution model). These models can be used to estimate the reproductive strategy of a species from a single sample of fish collected during the prespawning period to support species selection and data interpretation in environmental monitoring programs.

Regional and Long-Term Analyses of Stable Isotopes of Fish and Invertebrates Show Evidence of the Closure of a Pulp Mill and the Influence of Additional Stressors.

A bleached kraft pulp mill discharging effluent to the Mattagami River in northern Ontario, Canada, closed after almost 90 yr of operation. During its operation, effluent from the mill influenced biota in the downstream areas. To assess shifts in the reliance of biota from mill-derived nutrients, the isotopic composition (δ13 C and δ15 N) of white sucker (Catostomus commersoni) muscle and whole mayflies (Hexagenia sp.) were compared before (1990s) and after the pulp mill's closure (2012-2014). To better understand other potential sources of spatial and temporal change, samples from 3 other tributaries in the basin with dams, ongoing pulp mill operations, sites receiving sewage, and at several reference sites were collected and compared. Irrespective of time period, biota collected at sites downstream of both dams and active pulp mills tended to have elevated δ13 C values, but variable changes in δ15 N (negligible in most cases) when compared with upstream samples. The isotopic composition of mayflies varied at reference sites over time, with decreasing values of δ13 C and δ15 N (mayflies only) with increasing depth, and there was evidence of lower δ13 C in fish after the pulp mill closure. Overall, these results suggest the importance of long-term, regional-scale measurements for documenting the effects of stressors on nutrient use by aquatic species. Environ Toxicol Chem 2020;39:1207-1218. © 2020 SETAC.

Twenty years of transcriptomics, 17alpha-ethinylestradiol, and fish.

In aquatic toxicology, perhaps no pharmaceutical has been investigated more intensely than 17alpha-ethinylestradiol (EE2), the active ingredient of the birth control pill. At the turn of the century, the fields of comparative endocrinology and endocrine disruption research witnessed the emergence of omics technologies, which were rapidly adapted to characterize potential hazards associated with exposures to environmental estrogens, such as EE2. Since then, significant advances have been made by the scientific community, and as a result, much has been learned about estrogen receptor signaling in fish from environmental xenoestrogens. Vitellogenin, the egg yolk precursor protein, was identified as a major estrogen-responsive gene, establishing itself as the premier biomarker for estrogenic exposures. Omics studies have identified a plethora of estrogen responsive genes, contributing to a wealth of knowledge on estrogen-mediated regulatory networks in teleosts. There have been ~40 studies that report on transcriptome responses to EE2 in a variety of fish species (e.g., zebrafish, fathead minnows, rainbow trout, pipefish, mummichog, stickleback, cod, and others). Data on the liver and testis transcriptomes dominate in the literature and have been the subject of many EE2 studies, yet there remain knowledge gaps for other tissues, such as the spleen, kidney, and pituitary. Inter-laboratory genomics studies have revealed transcriptional networks altered by EE2 treatment in the liver; networks related to amino acid activation and protein folding are increased by EE2 while those related to xenobiotic metabolism, immune system, circulation, and triglyceride storage are suppressed. EE2-responsive networks in other tissues are not as comprehensively defined which is a knowledge gap as regulated networks are expected to be tissue-specific. On the horizon, omics studies for estrogen-mediated effects in fish include: (1) Establishing conceptual frameworks for incorporating estrogen-responsive networks into environmental monitoring programs; (2) Leveraging in vitro and computational toxicology approaches to identify chemicals associated with estrogen receptor-mediated effects in fish (e.g., male vitellogenin production); (3) Discovering new tissue-specific estrogen receptor signaling pathways in fish; and (4) Developing quantitative adverse outcome pathway predictive models for estrogen signaling. As we look ahead, research into EE2 over the past several decades can serve as a template for the array of hormones and endocrine active substances yet to be fully characterized or discovered.

Development of environmental effects monitoring protocol in Brazil: a fish guide study of three river estuaries.

In Brazil, there are no unified and effective environmental monitoring models for bodies of water. Thus, several methodologies are used that result in information that is often difficult to compare, especially for stakeholders involved in regional water management. Studies in some countries such as Australia, Chile, the USA, and Sweden use the monitoring model implemented in Canada that was developed in the early 1990s. This model was designed to evaluate whether the current environmental regulations are sufficiently protective for pulp and paper effluents and for metal mining effluents. In this study, the Canadian Environmental Effects Monitoring methodologies were applied to three different Brazilian river basins, with the goal of constructing a framework for monitoring environmental effects. Pilot studies were carried out in the estuarine regions of the Benevente, Jucu, and Santa Maria da Vitória river basins, which are important rivers in the state of Espírito Santo. Evaluations included fish health, bioaccumulation studies, benthic invertebrate survey, and physical-chemical analyses of water and sediment. The quality of the environments was evaluated by means of seasonal samplings and comparisons between discharge, upstream, and downstream areas. This study made it possible to identify appropriate fish species to be used in environmental effects monitoring and the environmental quality of the rivers themselves as well as knowledge and policy gaps to implement such monitoring programs in Brazil. The study raises questions about the adequacy of Brazilian environmental legislation concerning tidal rivers.

Developing Triggers for Environmental Effects Monitoring Programs for Trout-Perch in the Lower Athabasca River (Canada).

Baseline variations in trout-perch energy use (growth, gonad size) and energy storage (condition, liver size) were characterized between 2009 and 2015 in 8 reaches of the Athabasca River (AB, Canada), including 2 reaches upstream of the city of Fort McMurray (AB, Canada) and 6 reaches downstream of Fort McMurray among existing oil sands operations. Generalized linear models, used to account for background variation, indicated that fork length, gonad size, and liver size decreased, whereas body weight increased, in relation to river discharge, for both male and female trout-perch. Air temperature was positively correlated with liver size and negatively correlated with gonad size and body weight for females, but only positively correlated with gonad weight for males. These linear models explained approximately 20 to 25% of the variation in adjusted body size, and upward of 80% of the variation in adjusted body weight, liver weight, and gonad weight. Residuals from linear models were used to estimate normal ranges of variation for each of the fish population performance measures. Combined, the models and normal ranges can be used to assess subsequent monitoring data, providing potential triggers for follow-up monitoring activities. Environ Toxicol Chem 2019;38:0-0. Environ Toxicol Chem 2019;38:1890-1901. © 2019 SETAC.

Overview of Existing Science to Inform Oil Sands Process Water Release: A Technical Workshop Summary.

The extraction of oil sands from mining operations in the Athabasca Oil Sands Region uses an alkaline hot water extraction process. The oil sands process water (OSPW) is recycled to facilitate material transport (e.g., ore and tailings), process cooling, and is also reused in the extraction process. The industry has expanded since commercial mining began in 1967 and companies have been accumulating increasing inventories of OSPW. Short- and long-term sustainable water management practices require the ability to return treated water to the environment. The safe release of OSPW needs to be based on sound science and engineering practices to ensure downstream protection of ecological and human health. A significant body of research has contributed to the understanding of the chemistry and toxicity of OSPW. A multistakeholder science workshop was held in September 2017 to summarize the state of science on the toxicity and chemistry of OSPW. The goal of the workshop was to review completed research in the areas of toxicology, chemical analysis, and monitoring to support the release of treated oil sands water. A key outcome from the workshop was identifying research needs to inform future water management practices required to support OSPW return. Another key outcome of the workshop was the recognition that methods are sufficiently developed to characterize chemical and toxicological characteristics of OSPW to address and close knowledge gaps. Industry, government, and local indigenous stakeholders have proceeded to utilize these insights in reviewing policy and regulations. Integr Environ Assess Manag 2019;15:519-527. © 2019 SETAC.

An Adaptive Environmental Effects Monitoring Framework for Assessing the Influences of Liquid Effluents on Benthos, Water, and Sediments in Aquatic Receiving Environments.

Environmental effects monitoring (EEM) has been traditionally used to evaluate the effects of existing facilities discharging liquid effluents into natural receiving waters in Canada. EEM also has the potential to provide feedback to an ongoing project in an adaptive management context and can inform the design of future projects. EEM, consequently, can and should also be used to test the predictions of effects related to new projects. Despite EEM's potential for widespread applicability, challenges related to the effective implementation of EEM include the use of appropriate study designs and the adoption of tiers for increasing or decreasing monitoring intensity. Herein we describe a template for designing and implementing a 6-tiered EEM program that utilizes information from the project-planning and predevelopment baseline data collection stages to build on forecasts from the initial environmental impact assessment project-design stage and that feeds into an adaptive management process. Movement between the 6 EEM tiers is based on the exceedance of baseline monitoring triggers, forecast triggers, and management triggers at various stages in the EEM process. To distinguish these types of triggers, we review the historical development of numeric and narrative triggers as applied to chemical (water and sediment) and biological (plankton, benthos, fish) endpoints. We also provide an overview of historical study design issues and discuss how the 6 EEM tiers and associated triggers influence the temporal-spatial experimental design options and how the information gained through EEM could be used in an adaptive management context. Integr Environ Assess Manag 2018;14:552-566. © 2018 SETAC.

Developing and applying control charts to detect changes in water chemistry parameters measured in the Athabasca River near the oil sands: A tool for surveillance monitoring.

Control charting is a simple technique to identify change and is well suited for use in water quality programs. Control charts accounting for covariation associated with discharge and in some cases time were used to explore example and representative variables routinely measured in the Athabasca River near the oil sands area for indications of change. The explored variables include 5 major ions (chloride, sodium, sulfate, calcium, magnesium), 5 total metals (aluminum, iron, thallium, molybdenum, vanadium), and total suspended solids at two sites straddling the developments north of Fort McMurray. Regression equations developed from reference data (1988-2009) were used to predict observations and calculate residuals from later test data (2010-2016). Evidence of change was sought in the deviation of residual errors from the test period compared with the patterns expected and defined from probability distributions of the reference residuals using the odds ratio. In most cases, the patterns in test residuals were not statistically different from those expected from the reference period at either site, especially when data were examined annually. However, differences were found at both locations, more were found at the downstream site, and more differences emerged as data accumulated and were analyzed over time. In sum, the analyses at the downstream site suggest higher concentrations than predicted in most major ions, but the source of the changes is uncertain. In contrast, the concentrations of most metals at during the test period were lower than expected, which may be related to deposition patterns of materials or weathering of minerals during construction activities of the 2000s which influence the reference data used. The analyses also suggest alternative approaches may be necessary to understand change in some variables. Despite this, the results support the use of control charts to detect changes in water chemistry parameters and the value of the tool in surveillance phases of long-term and adaptive monitoring programs. Environ Toxicol Chem 2018;37:2296-2311. © 2018 SETAC.

Estrogen-responsive gene networks in the teleost liver: What are the key molecular indicators?

An overarching goal of environmental genomics is to leverage sensitive suites of markers that are robust and reliable to assess biological responses in a range of species inhabiting variable environments. The objective of this study was to identify core groups of transcripts and molecular signaling pathways that respond to 17alpha-ethylinestadiol (EE2), a ubiquitous estrogenic contaminant, using transcriptome datasets generated from six independent laboratories. We sought to determine which biomarkers and gene networks were those most robust and reliably detected in multiple laboratories. Six laboratories conducted microarray analysis in pieces of the same liver from male fathead minnows exposed to ∼15ng/L EE2 for 96h. There were common transcriptional networks identified in every dataset. These included down-regulation of gene networks associated with blood clotting, complement activation, triglyceride storage, and xenobiotic metabolism. Noteworthy was that more than ∼85% of the gene networks were suppressed by EE2. Leveraging both these data and those mined from the Comparative Toxicogenomics Database (CTD), we narrowed in on an EE2-responsive transcriptional network. All transcripts in this network responded ∼±5-fold or more to EE2, increasing reliability of detection. This network included estrogen receptor alpha, transferrin, myeloid cell leukemia 1, insulin like growth factor 1, insulin like growth factor binding protein 2, and methionine adenosyltransferase 2A. This estrogen-responsive interactome has the advantage over single markers (e.g. vitellogenin) in that these entities are directly connected to each other based upon evidence of expression regulation and protein binding. Thus, it represents an interacting functional suite of estrogenic markers. Vitellogenin, the gold standard for estrogenic exposures, can show high individual variability in its response to estrogens, and the use of a multi-gene approach for estrogenic chemicals is expected to improve sensitivity. In our case, the coefficient of variation was significantly lowered by the gene network (∼67%) compared to Vtg alone, supporting the use of this transcriptional network as a sensitive alternative for detecting estrogenic effluents and chemicals. We propose that screening chemicals for estrogenicity using interacting genes within a defined expression network will improve sensitivity, accuracy, and reduce the number of animals required for endocrine disruption assessments.

Using normal ranges for interpreting results of monitoring and tiering to guide future work: A case study of increasing polycyclic aromatic compounds in lake sediments from the Cold Lake oil sands (Alberta, Canada) described in Korosi et al. (2016).

Since the publishing of the Kelly et al. papers (2009, 2010) describing elevated contaminants in snow near the Alberta oil sands, there has been a significant expansion of monitoring efforts, enhanced by $50M a year contributed by industry to a regional Joint Oil Sands Monitoring (JOSM) program. In parallel to the intensification of research and monitoring efforts, including expansion of measured indicators, techniques for chemical analysis have also become more sensitive. Both factors contribute to the increased sensitivity and power, and improve our capacity to detect any change. The increase in capability requires a counterbalance to account for trivial change. This can be done using an interpretative approach that requires contextualization of differences to meaningfully inform environmental monitoring programs and provide focus for action. Experience obtained through 25 years of involvement with Canada's Environmental Effects Monitoring (EEM) program has shown that a tiered program informed by triggers can provide the context to make decisions about monitoring priorities. Here we provide a potential interpretation framework using a case study around the Korosi et al. (2016) study which found recent increases in alkylated polycyclic aromatic compounds (aPACs) in the Cold Lake in situ oil sands area. Public contaminant profiles from the JOSM studies in the oil sands region are used to evaluate the changes using an interpretation framework based on estimated normal ranges using existing data for site-specific, local and regional (distant) levels that was modelled after the tiered Canadian EEM design.

Using adaptive processes and adverse outcome pathways to develop meaningful, robust, and actionable environmental monitoring programs.

The primary goals of environmental monitoring are to indicate whether unexpected changes related to development are occurring in the physical, chemical, and biological attributes of ecosystems and to inform meaningful management intervention. Although achieving these objectives is conceptually simple, varying scientific and social challenges often result in their breakdown. Conceptualizing, designing, and operating programs that better delineate monitoring, management, and risk assessment processes supported by hypothesis-driven approaches, strong inference, and adverse outcome pathways can overcome many of the challenges. Generally, a robust monitoring program is characterized by hypothesis-driven questions associated with potential adverse outcomes and feedback loops informed by data. Specifically, key and basic features are predictions of future observations (triggers) and mechanisms to respond to success or failure of those predictions (tiers). The adaptive processes accelerate or decelerate the effort to highlight and overcome ignorance while preventing the potentially unnecessary escalation of unguided monitoring and management. The deployment of the mutually reinforcing components can allow for more meaningful and actionable monitoring programs that better associate activities with consequences. Integr Environ Assess Manag 2017;13:877-891. © 2017 The Authors. Integrated Environmental Assessment and Management Published by Wiley Periodicals, Inc. on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

How consistent are we? Interlaboratory comparison study in fathead minnows using the model estrogen 17α-ethinylestradiol to develop recommendations for environmental transcriptomics.

Fundamental questions remain about the application of omics in environmental risk assessments, such as the consistency of data across laboratories. The objective of the present study was to determine the congruence of transcript data across 6 independent laboratories. Male fathead minnows were exposed to a measured concentration of 15.8 ng/L 17α-ethinylestradiol (EE2) for 96 h. Livers were divided equally and sent to the participating laboratories for transcriptomic analysis using the same fathead minnow microarray. Each laboratory was free to apply bioinformatics pipelines of its choice. There were 12 491 transcripts that were identified by one or more of the laboratories as responsive to EE2. Of these, 587 transcripts (4.7%) were detected by all laboratories. Mean overlap for differentially expressed genes among laboratories was approximately 50%, which improved to approximately 59.0% using a standardized analysis pipeline. The dynamic range of fold change estimates was variable between laboratories, but ranking transcripts by their relative fold difference resulted in a positive relationship for comparisons between any 2 laboratories (mean R2 > 0.9, p < 0.001). Ten estrogen-responsive genes encompassing a fold change range from dramatic (>20-fold; e.g., vitellogenin) to subtle (∼2-fold; i.e., block of proliferation 1) were identified as differentially expressed, suggesting that laboratories can consistently identify transcripts that are known a priori to be perturbed by a chemical stressor. Thus, attention should turn toward identifying core transcriptional networks using focused arrays for specific chemicals. In addition, agreed-on bioinformatics pipelines and the ranking of genes based on fold change (as opposed to p value) should be considered in environmental risk assessment. These recommendations are expected to improve comparisons across laboratories and advance the use of omics in regulations. Environ Toxicol Chem 2017;36:2593-2601. © 2017 SETAC.