Collecting, Evaluating, and Communicating Ecotoxicology Data: ECOTOX Knowledgebase

The ECOTOXicology Knowledgebase (ECOTOX) has been locating and curating ecologically relevant toxicity data for over 30 years and is now a nationally and internationally recognized source of single-chemical toxicity test results for aquatic and terrestrial organisms. ECOTOX uses systematic and transparent procedures to conduct comprehensive literature searches, screen references, apply acceptance criteria, and extract all pertinent study and effects information. This lecture will allow viewers to deepen their knowledge of how the ECOTOX knowledgebase collects data, and downstream applications for EPA program offices, states, tribes, and other federal agencies. Additionally, attendees will learn to query and export data from ECOTOX. The session will begin with a short lecture on the background of ECOTOX and the ECOTOX data pipeline, a short example of new ECOTOX functions, then open up for users to ask questions. This abstract does not necessarily reflect US EPA policy.

1. Collecting, Evaluating, and Communicating Ecotoxicology Data: ECOTOX Knowledgebase Alexander R. Cole US EPA ORD Center for Computational Toxicology and Exposure Great Lakes Toxicology and Ecology Division Duluth, MN Center for Computational Toxicology and Exposure The views expressed in this presentation are those of the authors and do not necessarily reflect the views or policies of the U.S. EPA.

2. Ecological risk assessors need cost-effect methods to locate and evaluate high-quality ecological toxicity data Background and History 2 Ecological Risk Assessment for chemical registration and re-registration (USEPA Office of Pesticide Programs) Ambient Water Quality Criteria for Aquatic Life (USEPA Office of Water) Ecological Site Assessments and in Emergency Response Ecological hazard data for the Prioritization and Assessment of Chemicals for Toxic Substances Control Act/Lautenberg Act (USEPA Office of Pollution Prevention and Toxics) (USEPA Office of Land and Emergency Management - Superfund and Resource Conservation and Recovery Act; Regions, States, and Tribes).

3. Ecological risk assessors need cost-effect methods to locate high-quality ecological toxicity data Background and History 3 C haracterize ecological effects associated with a stressor (i.e., chemical) Develop chemical concentration goals to protect: surface water for aquatic life use ecological receptors that commonly come into contact with and/or consume biota that live in or on soil. Toxicity Reference Values (TRVs) Aquatic Life Criteria Environmental Screening Levels Benchmarks Screening Criteria or Benchmarks Soil Screening Levels Cleanup Target Levels Ecological Receptors (examples) Aquatic : fish, amphibians, benthic invertebrates, aquatic plants Terrestrial: plants, soil invertebrates, mammals, birds, amphibians, reptiles https://www.epa.gov/ecobox/epa-ecobox-tools-effects

4. Ecological risk assessors need cost-effect methods to locate high-quality ecological toxicity data Background and History 4 https://www.epa.gov/ecobox/epa-ecobox-tools-effects C haracterize ecological effects associated with a stressor (i.e., chemical) Relationship and causality based on laboratory studies (and some field or observational studies): Short-term (acute) and long-term (chronic) chemical exposures Measured response to varying chemical doses/concentrations Mortality Reduction in growth Reproductive impairment Changes in population size or numbers of species Bioaccumulation of chemical in organisms Disruption of community and ecosystem-level functions

5. Ecological risk assessors need cost-effect methods to locate high-quality ecological toxicity data Background and History 5 C haracterize ecological effects associated with a stressor (i.e., chemical) Relationship and causality based on laboratory studies (and some field or observational studies): Short-term (acute) and long-term (chronic) chemical exposures Controlled experiments with documentation/measurement of exposure conditions, control performance, responses Test Guidelines (examples) USEPA Series 850 - Ecological Effects Test Guidelines OECD Series on Testing and Assessment: Ecotoxicity Testing ASTM Guide for Conducting Acute Toxicity Tests Biological Test Methods - Canada PEER-REVIEWED PUBLICATIONS GOVERNMENT & INDUSTRY REPORTS

6. Ecological risk assessors need cost-effect methods to locate high-quality ecological toxicity data US EPA developed ecological toxicity databases AQUatic toxicity Information Retrieval (AQUIRE) database (Duluth, MN lab) PHYTOTOX (Corvallis, OR lab) TERRETOX (Corvallis, OR lab) 6 Background and History 6 MULTIPLE PUBLICATIONS DIVERSITY OF SPECIES

7. Systematic and transparent protocols developed over 30 years: Conduct comprehensive literature searches for toxicity data in the peer-reviewed and grey literature Review applicability of studies following established criteria Extract relevant study and toxicity results into a structured database Accessible, structured empirical data from in vivo toxicity tests On-going chemical-based literature searches Monthly review of recent ecotoxicology publications Toxicity data provided to US EPA Programs, Regions, and researchers Updated quarterly to public website (www.epa.gov/ecotox) 7 What is the ECOTOXicology Knowledgebase?

8. What is the ECOTOXicology Knowledgebase? www.epa.gov/ecotox 8 ECOTOX Overview: Olker et al. 2022   https://doi.org/10.1002/etc.5324

9. ECOTOX Pipeline 9 Conduct literature searches Chemical verification & search term development Identify & acquire potentially applicable studies Review literature for applicability Extract study and toxicity data Provide toxicity results and study details Planning and Identification Screening Eligibility Included Title/Abstract Screening Full Text Review Literature Search Use chemical-specific search terms to query multiple literature search engines. Chemical-based Search Terms Chemical name and CASRN Synonyms, tradenames Other relevant forms Data Extraction ECOTOX-specific Controlled Vocabularies Test chemical Test organism Study methods and test conditions Toxicity results Updated to public website, with downloadable outputs Established applicability (inclusion) criteria Documentation of exclusion reason

10. Verify CASRN Search various sources for chemical terms STN Pesticide Action Network EPA’s Pesticide Fate Database EPA’s Chemicals Dashboard Synonyms Eliminate poor search terms Develop search string Tak(Acilid OR Albrass OR Bexton OR "CP 31393" OR "Kartex A" OR Muharicid OR Niticid OR Propachlor OR Propachlore OR Ramrod OR Satecid OR "US EPA PC Code 019101") Chemical-based Search Terms ECOTOX Pipeline Conduct literature searches Chemical verification & search term development Identify & acquire potentially applicable studies Review literature for applicability Extract study and toxicity data Provide toxicity results and study details Planning and Identification Screening Eligibility Included 10

11. Title/Abstract Screening Full Text Review Established applicability (inclusion) criteria which can be expressed as PECO statement Documentation of exclusion reason ECOTOX Pipeline Conduct literature searches Chemical verification & search term development Identify & acquire potentially applicable studies Review literature for applicability Extract study and toxicity data Provide toxicity results and study details Planning and Identification Screening Eligibility Included 11

12. Key Area Data Requirement P (Population) Species Taxonomically verifiable, ecologically-relevant organisms (including cells, organs, gametes, embryos, plant cuttings) [NOT bacteria, humans, monkeys, viruses, or yeast] E (Exposure) Chemical Single, verifiable chemical toxicants, administered through an acceptable route. Exposure Amount (Concentration) Exposure amount is quantified, either as a concentration in the environment when administered via soil or water, or as a dosage when introduced directly into or on the organism, via injection, orally, or topically. Exposure Duration Known duration from the time of initial exposure to the time of measurement. C (Comparator/ Control) Control Must have a control treatment O (Outcome) Effect Biological effect measured Effect concurrent with associated chemical exposure Publication Type Primary source of the data [NOT a Review] Study must be a full article in English Inclusion Criteria 12 Identify and acquire potentially applicable studies Review literature for applicability Adapted from Olker et al. 2022  

13. All Excluded and Non-Applicable studies are Tagged with the reason for rejection Abstract – published as an abstract Bacteria – only test organism is a bacteria CAS # Unavailable – could not verify/locate chemical CAS Registry number Chemical method – description of chemical analysis procedures Fate – only report chemical distribution in media Human Health – data on human subjects of surrogate animal subjects for human health risk assessment Incident – reports death of animal by poison, but does not provide concentration/duration of exposure Method – paper only reports methods for conducting a toxicity test or other aspect of an experiment Mixture – paper reports results from mixture of chemicals; no single chemical exposure results Modeling – results of the development of a model; no primary data available No Conc – the authors report a response in an organism but do not provide conc/dose/app rate No Duration – duration of exposure is not presented No Effect – paper does not report observed responses adverse of otherwise No Toxicant (ozone, CO2) Non-English Nutrient – in situ chemical tested as nutrient PUBL AS – duplicate data published elsewhere Retracted – paper retracted by Journal Review – primary data published elsewhere Sediment – only sediment concentration presented Survey – chemical measured in organism, but lack quantification of exposure (dose/duration) Virus – virus is only test organism Yeast – yeast is only test organism Exclusion Documentation 13

14. NOEC = No Observed Effect Level LOEC = Lowest Observed Effect Level NR = Not Reported Data Extraction Example of multiple ECOTOX records from a single study: ECOTOX Pipeline Conduct literature searches Chemical verification & search term development Identify & acquire potentially applicable studies Review literature for applicability Extract study and toxicity data Provide toxicity results and study details Planning and Identification Screening Eligibility Included 14

15. ECOTOX Data Fields 15 Unique Identifiers Chemical ID CASRN DTXSID Taxonomic ID NCBI TaxID ITIS TSN Extract study and toxicity data

16. Chemicals 16 Pesticides PFAS 12,982 compounds

17. Species (n = 13,976) 17 % of Records by Species Group Decadal Shifts in Test Species

18. Diversity of Effects 18 % of Records by Effect Group Increases in Biochemical and Genetic Effects Reported

19. EPA Program Offices and Regions, States, Tribes, Other Federal Agencies and International Entities Chemical environmental toxicity data for aquatic and terrestrial organisms Applications Provides data to 19 Ecological Risk Assessments Ambient Water Quality Criteria Ecological Screening Values Chemical Prioritization Emergency Response

20. EPA Program Offices and Regions, States, Tribes, Other Federal Agencies and International Entities Chemical environmental toxicity data for aquatic and terrestrial organisms Applications Tools and Applications Species Sensitivity Distributions (e.g., US EPA’s SSD Toolbox, Endangered species analyses, U.S. EPA’s WebICE , NOAA’s CAFE ) Predicted No Effect Concentrations (PNECs) and Eco-TTCs (e.g., EnviroTox , NORMAN) Quantitative Structure-Activity Relationships and other models Adverse Outcome Pathway (AOP) development Identify data gaps, Inform study design, Compare to new toxicity studies Linking environmental contaminant concentrations to potential effects Toxicity Reference Values (TRVs) and Benchmarks Provides data to Data used for 20 https://www.epa.gov/chemical-research/interim-ecological-soil-screening-level-documents https://doi.org/10.1002/etc.4382 https://www.norman-network.com/nds/ecotox https://doi.org/10.23645/epacomptox.11971392 https://qsartoolbox.org https://www.epa.gov/tsca-screening-tools/ecological-structure-activity-relationships-ecosar-predictive-model https://www.epa.gov/chemical-research/toxicity-estimation-software-tool-test T.E.S.T. (Toxicity Estimation Software Tool)

21. Health & Environmental Research Online (HERO)* (https://hero.epa.gov/) EPA Program Offices and Regions, States, Tribes, Other Federal Agencies and International Entities Chemical environmental toxicity data for aquatic and terrestrial organisms Applications Tools and Applications Databases/Resources Data linked to U.S. EPA CompTox Chemicals Dashboard (https://comptox.epa.gov/dashboard/) OECD eChemPortal (https://www.echemportal.org/echemportal/ Adverse Outcome Pathway Wiki* (https://aopwiki.org/) Sequence Alignment to Predict Across Species Susceptibility ( SeqAPASS )* (https://doi.org/10.1093/toxsci/kfw119 ) *Future Biological Ontologies* Pathways and Disease Databases* 21 http://www.obofoundry.org/ https://lod-cloud.net/ Provides data to Data used for

22. Example 22

23. 23 6PPD-quinone https://www.science.org/content/article/common-tire-chemical-implicated-mysterious-deaths-risk-salmon https://www.science.org/doi/epdf/10.1126/science.abd6951

24. 24 6PPD-quinone https://www.science.org/doi/epdf/10.1126/science.abd6951 TWP = tire tread wear particle

25. Identify and curate relevant ecotoxicity studies on tire rubber antioxidant 6PPD and its transformation product 6PPD-quinone Targeted literature searches Rapid identification of recent publications Monthly reviews Consultation OW & Regions 25 6PPD-quinone Identification and curation of ecotoxicity results Lit Search # Refs reviewed # Refs in ECOTOX April 2021 6,129 2 Sept 2022 ‘Refresh’ 1,264 21 Sept 2023 ‘Refresh’ 2,364 36 June 2024 ‘Refresh’ 539 44 + More Pending www.epa.gov/ecotox

26. Problem Statement : Toxicity data of 6PPD-quinone on aquatic organisms needed for evaluation of toxicity effects, species sensitivity, and determination of mode of action. 6PPD-quinone

27. 27 6PPD-quinone

28. 28 6PPD-quinone

29. 29 6PPD-quinone

30. 30 6PPD-quinone

31. 31 6PPD-quinone

32. 32 6PPD-quinone

33. Summary 33

34. 1 ECOTOXicology Knowledgebase was developed for and continues to support chemical risk assessment and research Protocols and vocabulary evolved over 30+ year history Meet the demands for increased pace of chemical assessments Expand to reflect shifts in toxicity testing paradigm Provides detailed study and toxicity result details in standard, structured format for evaluation and analysis Dependent on author-reported information Curated data on public website ( www.epa.gov/ecotox ), readily available for exploration, querying, and export for risk assessments, risk management and research Summary 34

35. Acknowledgements and Contact Info 35 Contract staff: General Dynamics Information Technology (GDIT) SpecPro Professional Services (SPS) Senior Environmental Employment (SEE) staff Alex Cole [email protected] Jennifer Olker [email protected] Dale Hoff [email protected] ECOTOX Support [email protected] More info on 6PPD-Q: https://www.epa.gov/chemical-research/6ppd-quinone