Increasing Scientific Confidence in Adverse Outcome Pathways: Application of Tailored Bradford-Hill Considerations for Evaluating Weight of Evidence.

Systematic consideration of scientific support is a critical element in developing and, ultimately, using adverse outcome pathways (AOPs) for various regulatory applications. Though weight of evidence (WoE) analysis has been proposed as a basis for assessment of the maturity and level of confidence in an AOP, methodologies and tools are still being formalized. The Organization for Economic Co-operation and Development (OECD) Users' Handbook Supplement to the Guidance Document for Developing and Assessing AOPs (OECD 2014a; hereafter referred to as the OECD AOP Handbook) provides tailored Bradford-Hill (BH) considerations for systematic assessment of confidence in a given AOP. These considerations include (1) biological plausibility and (2) empirical support (dose-response, temporality, and incidence) for Key Event Relationships (KERs), and (3) essentiality of key events (KEs). Here, we test the application of these tailored BH considerations and the guidance outlined in the OECD AOP Handbook using a number of case examples to increase experience in more transparently documenting rationales for assigned levels of confidence to KEs and KERs, and to promote consistency in evaluation within and across AOPs. The major lessons learned from experience are documented, and taken together with the case examples, should contribute to better common understanding of the nature and form of documentation required to increase confidence in the application of AOPs for specific uses. Based on the tailored BH considerations and defining questions, a prototype quantitative model for assessing the WoE of an AOP using tools of multi-criteria decision analysis (MCDA) is described. The applicability of the approach is also demonstrated using the case example aromatase inhibition leading to reproductive dysfunction in fish. Following the acquisition of additional experience in the development and assessment of AOPs, further refinement of parameterization of the model through expert elicitation is recommended. Overall, the application of quantitative WoE approaches hold promise to enhance the rigor, transparency and reproducibility for AOP WoE determinations and may play an important role in delineating areas where research would have the greatest impact on improving the overall confidence in the AOP.

Weight of evidence needed to substantiate a health effect for probiotics and prebiotics: regulatory considerations in Canada, EU, and US

Successful and responsible introduction of probiotic and prebiotic products into the worldwide marketplace requires labelling for health benefits that meets consumer needs, adheres to regulatory standards and does not overextend scientific evidence. Regulations differ among countries, but underlying all is an emphasis on scientific credibility of any statements of health benefits. This paper considers the value of different types of evidence offered in substantiation of efficacy and reviews different regulatory approaches to labelling for health claims. Limitations of in vitro, animal and different types of human studies used for efficacy substantiation for probiotics and prebiotics are discussed.

Characterization of toxic impacts on living marine resources in tidal rivers of the Chesapeake Bay

In 1999, the Chesapeake Bay Program completed a survey of existing data on chemical contaminants and the potential for bioeffects in 38 tidal river systems of Chesapeake Bay. This review led to the identification of 20 areas for which there were insufficient data to adequately characterize the potential for contaminant bioeffects on the Bay’s living resources. The goal of the present study was to estimate the current status of ecological condition in five of these areas and thus help to complete the overall toxics inventory for the Bay. These five systems included the Chester River, Nanticoke River, Pocomoke River, Lower Mobjack Bay (Poquosin and Back Rivers) and the South and Rhode Rivers. This study utilized a Sediment Quality Triad (SQT) approach in combination with additional water-column contaminant analysis to allow for a “weight of evidence” assessment of environmental condition. A total of 60 stations distributed among the five systems, using a probabilistic stratified random design, were sampled during the summer of 2004 to allow for synoptic measures of sediment contamination, sediment toxicity, and benthic condition. Upon completion of all analyses, stations were assigned to one of four categories based on the three legs of the triad. Stations with high sediment quality had no hits on any of the three legs of the triad; those with moderate quality had one hit; those with marginal quality had two hits; and those with poor quality had hits for all three legs of the triad. The Pocomoke River had by far the largest proportion of the total area (97.5%) classified as having high sediment quality, while the Rhode/South system had the highest proportion (11.4%) classified as poor. None of the stations in the Chester River, Nanticoke River, and Lower Mobjack Bay systems were classified as poor. More than 65% of the area of each of the five systems was classified with high to moderate sediment quality. The Rhode/South system had 30.4% of total area classified with marginally to severely poor quality. The results of this study highlight the importance of using multiple indicators and a “weight of evidence” approach to characterize environmental quality and the potential bioeffects of toxic contaminants.

Ecological condition of coastal ocean and estuarine waters of the U.S. South Atlantic Bight: 2000-2004

In March-April 2004, the National Oceanic and Atmospheric Administration (NOAA), U.S. Environmental Protection Agency (EPA), and State of Florida (FL) conducted a study to assess the status of ecological condition and stressor impacts throughout the South Atlantic Bight (SAB) portion of the U.S. continental shelf and to provide this information as a baseline for evaluating future changes due to natural or human-induced disturbances. The boundaries of the study region extended from Cape Hatteras, North Carolina to West Palm Beach, Florida and from navigable depths along the shoreline seaward to the shelf break (~100m). The study incorporated standard methods and indicators applied in previous national coastal monitoring programs — Environmental Monitoring and Assessment Program (EMAP) and National Coastal Assessment (NCA) — including multiple measures of water quality, sediment quality, and biological condition. Synoptic sampling of the various indicators provided an integrative weight-of-evidence approach to assessing condition at each station and a basis for examining potential associations between presence of stressors and biological responses. A probabilistic sampling design, which included 50 stations distributed randomly throughout the region, was used to provide a basis for estimating the spatial extent of condition relative to the various measured indicators and corresponding assessment endpoints (where available). Conditions of these offshore waters are compared to those of southeastern estuaries, based on data from similar EMAP/NCA surveys conducted in 2000-2004 by EPA, NOAA, and partnering southeastern states (Florida, Georgia, South Carolina, North Carolina, Virginia) (NCA database for estuaries, EPA Gulf Ecology Division, Gulf Breeze FL). Data from a total of 747 estuarine stations are included in this database. As for the offshore sites, the estuarine samples were collected using standard methods and indicators applied in previous coastal EMAP/NCA surveys including the probabilistic sampling design and multiple indicators of water quality, sediment quality, and biological condition (benthos and fish). The majority of the SAB had high levels of DO in near-bottom water (> 5 mg L-1) indicative of "good" water quality. DO levels in bottom waters exceeded this upper threshold at all sites throughout the coastal-ocean survey area and in 76% of estuarine waters. Twenty-one percent of estuarine bottom waters had moderate levels of DO between 2 and 5 mg L-1 and 3% had DO levels below 2 mg L-1. The majority of sites with DO in the low range considered to be hypoxic (< 2 mg L-1) occurred in North Carolina estuaries. There also was a notable concentration of stations with moderate DO levels (2 – 5 mg L-1) in Georgia and South Carolina estuaries. Approximately 58% of the estuarine area had moderate levels of chlorophyll a (5-10 μg L-1) and about 8% of the area had higher levels, in excess of 10 μg L-1, indicative of eutrophication. The elevated chlorophyll a levels appeared to be widespread throughout the estuaries of the region. In contrast, offshore waters throughout the region had relatively low levels of chlorophyll a with 100% of the offshore survey area having values < 5 μg L-1.

Ecological condition of coastal ocean waters of the U.S. continental shelf off South Florida: 2007

A study was initiated with field work in May 2007 to assess the status of ecological condition and stressor impacts throughout the U.S. continental shelf off South Florida, focusing on soft-bottom habitats, and to provide this information as a baseline for evaluating future changes due to natural or human-induced disturbances. The boundaries of the study region extended from Anclote Key on the western coast of Florida to West Palm Beach on the eastern coast of Florida, inclusive of the Florida Keys National Marine Sanctuary (FKNMS), and from navigable depths along the shoreline seaward to the shelf break (~100m). The study incorporated standard methods and indicators applied in previous national coastal monitoring programs — U.S. Environmental Protection Agency’s (EPA) Environmental Monitoring and Assessment Program (EMAP) and National Coastal Assessment (NCA) — including multiple measures of water quality, sediment quality, and biological condition. Synoptic sampling of the various indicators provided an integrative weight-of-evidence approach to assessing condition at each station and a basis for examining potential associations between presence of stressors and biological responses. A probabilistic sampling design, which included 50 stations distributed randomly throughout the region, was used to provide a basis for estimating the spatial extent of condition relative to the various measured indicators and corresponding assessment endpoints (where available). The study was conducted through a large cooperative effort by National Oceanic and Atmospheric Administration (NOAA)/National Centers for Coastal Ocean Science (NCCOS), EPA, U.S. Geological Survey (USGS), NOAA/Oceanic and Atmospheric Research (OAR)/Atlantic Oceanographic and Meteorological Laboratory in Miami, FKNMS, and the Florida Fish and Wildlife Conservation Commission (FWC). The majority of the South Florida shelf had high levels of dissolved oxygen (DO) in near-bottom water (> 5 mg L-1) indicative of “good” water quality.. DO levels in bottom waters exceeded this upper threshold at 98.8% throughout the coastal-ocean survey area. Only 1.2% of the region had moderate DO levels (2-5 mg/L) and no part of the survey area had DO <2.0 mg/L. In addition, offshore waters throughout the region had relatively low levels of total suspended solids (TSS), nutrients, and chlorophyll a indicative of oligotrophic conditions. Results suggested good sediment quality as well. Sediments throughout the region, which ranged from sands to intermediate muddy sands, had low levels of total organic carbon (TOC) below bioeffect guidelines for benthic organisms. Chemical contaminants in sediments were also mostly at low, background levels. For example, none of the stations had chemicals in excess of corresponding Effects-Range Median (ERM) probable bioeffect values or more than one chemical in excess of lower-threshold Effects-Range Low (ERL) values. Cadmium was the only chemical that occurred at moderate concentrations between corresponding ERL and ERM values. Sixty fish samples from 28 stations were collected and analyzed for chemical contaminants. Eleven of these samples (39% of sites) had moderate levels of contaminants, between lower and upper non-cancer human-health thresholds, and ten (36% of sites) had high levels of contaminants above the upper threshold.

Harmonised framework for ecological risk assessment of sediments from ports and estuarine zones of North and South Atlantic

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A critical comparison of different approaches to sediment-quality assessments in the Santos estuarine system in Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Intra- and interlaboratory reliability of a cryopreserved trout hepatocyte assay for the prediction of chemical bioaccumulation potential

Measured rates of intrinsic clearance determined using cryopreserved trout hepatocytes can be extrapolated to the whole animal as a means of improving modeled bioaccumulation predictions for fish. To date, however, the intra- and interlaboratory reliability of this procedure has not been determined. In the present study, three laboratories determined in vitro intrinsic clearance of six reference compounds (benzo[a]pyrene, 4-nonylphenol, di-tert-butyl phenol, fenthion, methoxychlor and o-terphenyl) by conducting substrate depletion experiments with cryopreserved trout hepatocytes from a single source. O-terphenyl was excluded from the final analysis due to nonfirst-order depletion kinetics and significant loss from denatured controls. For the other five compounds, intralaboratory variability (% CV) in measured in vitro intrinsic clearance values ranged from 4.1 to 30%, while interlaboratory variability ranged from 27 to 61%. Predicted bioconcentration factors based on in vitro clearance values exhibited a reduced level of interlaboratory variability (5.3-38% CV). The results of this study demonstrate that cryopreserved trout hepatocytes can be used to reliably obtain in vitro intrinsic clearance of xenobiotics, which provides support for the application of this in vitro method in a weight-of-evidence approach to chemical bioaccumulation assessment.

Harmonised framework for ecological risk assessment of sediments from ports and estuarine zones of North and South Atlantic

This paper presents a harmonised framework of sediment quality assessment and dredging material characterisation for estuaries and port zones of North and South Atlantic. This framework, based on the weight-of-evidence approach, provides a structure and a process for conducting sediment/dredging material assessment that leads to a decision. The main structure consists of step 1 (examination of available data); step 2 (chemical characterisation and toxicity assessment); decision 1 (any chemical level higher than reference values? are sediments toxic?); step 3 (assessment of benthic community structure); step 4 (integration of the results); decision 2 (are sediments toxic or benthic community impaired?); step 5 (construction of the decision matrix) and decision 3 (is there environmental risk?). The sequence of assessments may be interrupted when the information obtained is judged to be sufficient for a correct characterisation of the risk posed by the sediments/dredging material. This framework brought novel features compared to other sediment/dredging material risk assessment frameworks: data integration through multivariate analysis allows the identification of which samples are toxic and/or related to impaired benthic communities; it also discriminates the chemicals responsible for negative biological effects; and the framework dispenses the use of a reference area. We demonstrated the successful application of this framework in different port and estuarine zones of the North (Gulf of Cadiz) and South Atlantic (Santos and Paranagua Estuarine Systems).

Sediment Quality in Puget Sound Year 3 - Southern Puget Sound

As a component of a three-year cooperative effort of the Washington State Department of Ecology and the National Oceanic and Atmospheric Administration, surficial sediment samples from 100 locations in southern Puget Sound were collected in 1999 to determine their relative quality based on measures of toxicity, chemical contamination, and benthic infaunal assemblage structure. The survey encompassed an area of approximately 858 km2, ranging from East and Colvos Passages south to Oakland Bay, and including Hood Canal. Toxic responses were most severe in some of the industrialized waterways of Tacoma’s Commencement Bay. Other industrialized harbors in which sediments induced toxic responses on smaller scales included the Port of Olympia, Oakland Bay at Shelton, Gig Harbor, Port Ludlow, and Port Gamble. Based on the methods selected for this survey, the spatial extent of toxicity for the southern Puget Sound survey area was 0% of the total survey area for amphipod survival, 5.7% for urchin fertilization, 0.2% for microbial bioluminescence, and 5- 38% with the cytochrome P450 HRGS assay. Measurements of trace metals, PAHs, PCBs, chlorinated pesticides, other organic chemicals, and other characteristics of the sediments, indicated that 20 of the 100 samples collected had one or more chemical concentrations that exceeded applicable, effects-based sediment guidelines and/or Washington State standards. Chemical contamination was highest in eight samples collected in or near the industrialized waterways of Commencement Bay. Samples from the Thea Foss and Middle Waterways were primarily contaminated with a mixture of PAHs and trace metals, whereas those from Hylebos Waterway were contaminated with chlorinated organic hydrocarbons. The remaining 12 samples with elevated chemical concentrations primarily had high levels of other chemicals, including bis(2-ethylhexyl) phthalate, benzoic acid, benzyl alcohol, and phenol. The characteristics of benthic infaunal assemblages in south Puget Sound differed considerably among locations and habitat types throughout the study area. In general, many of the small embayments and inlets throughout the study area had infaunal assemblages with relatively low total abundance, taxa richness, evenness, and dominance values, although total abundance values were very high in some cases, typically due to high abundance of one organism such as the polychaete Aphelochaeta sp. N1. The majority of the samples collected from passages, outer embayments, and larger bodies of water tended to have infaunal assemblages with higher total abundance, taxa richness, evenness, and dominance values. Two samples collected in the Port of Olympia near a superfund cleanup site had no living organisms in them. A weight-of-evidence approach used to simultaneously examine all three “sediment quality triad” parameters, identified 11 stations (representing 4.4 km2, 0.5% of the total study area) with sediment toxicity, chemical contamination, and altered benthos (i.e., degraded sediment quality), 36 stations (493.5 km2, 57.5% total study area) with no toxicity or chemical contamination (i.e., high sediment quality), 35 stations (274.1 km2, 32.0% total study area) with one impaired sediment triad parameter (i.e., intermediate/high sediment quality), and 18 stations (85.7km2, 10.0% total study area) with two impaired sediment parameters (i.e., intermediate/degraded quality sediments). Generally, upon comparison, the number of stations with degraded sediments based upon the sediment quality triad of data was slightly greater in the central Puget Sound than in the northern and southern Puget Sound study areas, with the percent of the total study area degraded in each region decreasing from central to north to south (2.8, 1.3 and 0.5%, respectively). Overall, the sediments collected in Puget Sound during the combined 1997-1999 surveys were among the least contaminated relative to other marine bays and estuaries studied by NOAA using equivalent methods. (PDF contains 351 pages)

Histological alterations in the Antarctic cod, Trematomus bernacchii from Davis Station, East Antarctica

During the summer season 2009/10, a comprehensive environmental impact assessment of the sewage discharge was conducted at Davis Station in the Vestfold Hills region of East Antarctica. As part of this project, a survey of the histology of liver, gill, gonad and muscle tissues in the Antarctic rock cod Trematomus bernacchii from nearshore sites in the receiving environment close to Davis Station in was completed. Fish from 4 sites were examined; 1 site adjacent to the Davis Station sewage outfall (within 500 m of the point of discharge), 2 sites approximately 2 km from the outfall (Anchorage Island and Antennae farm), and 1 site approximately 10 km away from the outfall and adjacent to an Adelie penguin population (Kazak Island). All fish sampled from the sewage outfall site exhibited significant histological alterations in all major tissues. Fish from the other 3 sites showed some alterations in either gill and/or liver tissues. Pathological abnormalities present in all fish collected near the sewage outfall included: extensive multifocal cysts of unknown etiology with necrotic liquification; multifocal granuloma with associated inflammation; coagulative necrosis in the liver; and lamellar hyperplasia with associated proliferation and lamella fusion of the gills. Results of this work form part of a weight of evidence approach alongside ecological monitoring, chemical analysis, ecotoxicological testing and dispersal modelling of the discharge plume which is being used to inform and direct upgrades to the Australian Antarctic Divisions operations and current sewage discharge practises at Davis Station.

Integrated sediment quality assessment in Paranagua Estuarine System, Southern Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Integrated sediment quality assessment in Paranagua Estuarine System, Southern Brazil

Sediment quality from Paranagua Estuarine System (PES), a highly important port and ecological zone, was evaluated by assessing three lines of evidence: (1) sediment physical-chemical characteristics; (2) sediment toxicity (elutriates, sediment-water interface, and whole sediment); and (3) benthic community structure. Results revealed a gradient of increasing degradation of sediments (i.e. higher concentrations of trace metals, higher toxicity, and impoverishment of benthic community structure) towards inner PES. Data integration by principal component analysis (PCA) showed positive correlation between some contaminants (mainly As, Cr, Ni, and Pb) and toxicity in samples collected from stations located in upper estuary and one station placed away from contamination sources. Benthic community structure seems to be affected by both pollution and natural fine characteristics of the sediments, which reinforces the importance of a weight-of-evidence approach to evaluate sediments of PES. (C) 2008 Elsevier Inc. All rights reserved.

Optimizing the aquatic toxicity assessment under REACH through an integrated testing strategy (ITS).

To satisfy REACH requirements a high number of data on chemical of interest should be supplied to the European Chemicals Agency. To organize the various kinds of information and help the registrants to choose the best strategy to obtain the needed information limiting at the minimum the use of animal testing, integrated testing strategies (ITSs) schemes can be used. The present work deals with regulatory data requirements for assessing the hazards of chemicals to the aquatic pelagic environment. We present an ITS scheme for organizing and using the complex existing data available for aquatic toxicity assessment. An ITS to optimize the choice of the correct prediction strategy for aquatic pelagic toxicity is described. All existing information (like physico-chemical information), and all the alternative methods (like in silico, in vitro or the acute-to-chronic ratio) are considered. Moreover the weight of evidence approach to combine the available data is included.