Ecotoxicology Knowledgebase

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ECOTOXicology Knowledgebase: Modernizing the Literature Review and Data Curation Processes, and Mapping Ecological Toxicity of Per- and Polyfluoroalkyl Substances (PFAS)

Jennifer OlkerPhoto, image Postdoctoral area measures 2” H x 6.93” W and can be Researchermasked by a collage strip of one, two or three images. Colleen ElonenThe photo, imageECOTOX area is located 3.19” from coordinator left and 3.81” from top of page. Each image used in collage should be reduced or cropped to a maximum of 2” high, stroked with a 1.5 pt white frame and positioned edge-to-edge with accompanying images. US EPA ECOTOX Project Team: Colleen Elonen Jennifer Olker GDIT contract staff Dale Hoff SEE staff Rong-Lin Wang Office of Research and Development www.epa.gov/ecotox Overview

• Background and History for ECOTOX Knowledgebase • Modernizing the ECOTOX Pipeline (C. Elonen, SOT 2020) • Mapping ecological toxicity of PFAS with ECOTOX Protocols (J. Olker, SOT 2020)

2 What is the ECOTOX Knowledgebase?

Publicly available, curated database providing toxicity data from single-chemical exposure studies to aquatic life, terrestrial plants, and wildlife

• From comprehensive search and review of open and grey literature – Data extracted from acceptable studies, with up to 250 fields – Updated quarterly

• 30+ year history: www.epa.gov/ecotox/ Originated in the early 1980s, US Environmental Protection Agency Office of Research and Development 3 Who uses the ECOTOX Knowledgebase?

Clients Contacting ECOTOX Support line 2005 - 2016 (n = 2813) US EPA Headquarters Local Govt. International Govt. 4% 1% 8% Govt. Contractors 25%

US EPA Regional Office 4% Unknown 5% University/Academia Other Federal 6% Agencies 4%

US EPA Laboratory 4% State Govt. 7%

Private Sector ~8000 Distinct hosts/month 32%

4 Program Offices & Regions Applications: use in environmental decision making

Used for every Ambient Water Quality Criteria for Aquatic Life since 1985.

Used for every Ecological Risk Assessment for Office of Pesticides for chemical registration and re-registration (FY19 – 30 chemicals).

Used by OLEM (Superfund and RCRA), HQ, Regions and States for site assessments and in emergency response

Providing ecological hazard data for the prioritization and assessment of chemicals for TSCA/Lautenberg Act

Providing ecological toxicity data for PFAS to researchers, EPA ERA Forum, DoD Tri-Services ERA Work Group, and others 5 Applications of ECOTOX

ECOTOX Knowledgebase Chemical EPA Program Offices and Regions, environmental States, Tribes, Other Federal Agencies toxicity data for and International Entities aquatic life, Ecological Risk Assessments terrestrial plants Ambient Water Quality Criteria and wildlife Ecological Screening Values Chemical Prioritization Emergency Response

Tools and Applications Species Sensitivity Distributions (e.g., US EPA’s WebICE, NOAA’s CAFÉ) PNECs and threshold values (e.g., EcoTTC) QSAR (e.g., ECOSAR, TEST, OECD QSAR Toolbox) BCF modeling and validation Chemicals SeqAPASS HERO Adverse Outcome Pathway (AOP) development Dashboard (Health & Environmental Research Online)

Interoperability Interoperability with databases/tools 6 ECOTOX Pipeline: Systematic Review/Data Curation

Chemical verification Chemical-based Search Terms: Sources include: • Chemical name and CASRN • STN and development of • Synonyms, tradenames • Pesticide Action Network (PAN) search terms • Other relevant forms (metabolites, degradants, • EPA’s Pesticide Fate Database (PFATE) parent compound, related chemicals) • EPA’s Chemistry Dashboard.

Literature search: Use chemical-specific search terms to query multiple literature search engines Citations from: ProQuest/ Science ToxNet Dissertation Agricola Current Contents Already in CSA Direct Abstracts (WoS) Unify* Conduct literature n = 5,631 11,178 317 234 4,861 15,347 333 Identification searches *Internal USEPA ECOTOX database

~37,000 citations downloaded Initial removal of duplicates

Not applicable (excluded): n = 8,265 references Identify and acquire Title and Abstract Screening Chem Methods: 3,462 No Toxicant: 221 potentially applicable n = 8,653 references Human Health: 1,797 Duplicate: 153 False Hit: 1,333 Review: 50 studies Fate: 510 Mixture: 12 Screening Survey: 287 Other: 121 Bacteria: 233 Review literature for For Review (Full Text Screening) No PFAS in reference: n = 85 references applicability to n = 388 references Did not meet acceptability criteria (excluded): ECOTOX n = 142 references Eligibility

Awaiting Review and Data Extraction Data Extracted from Acceptable n = 1 references Extract data into Papers ECOTOX n = 245 references with 7,496 total records

Included Knowledgebase 7 ECOTOX Pipeline: Systematic Review/Data Curation

~37,000 citations downloaded Initial removal of duplicates

Data Extracted from Acceptable Papers Awaiting Review and Data Extraction Data extraction n = 245 references with 7,496 total records n = 1 references

Study quality Data from High Quality Studies Included evaluation n = ___ references 8 Identify, Test, and QA Search Terms

Chemical verification Search various sources for chemical terms, and development of Synonyms, verify CAS, eliminate poor search terms search terms

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") Conduct literature Identification searches

Identify and acquire Couple hour process potentially applicable studies Screening

Review literature for applicability to ECOTOX Eligibility

Enter chemical terms Data extraction into template for Study quality abstracting databases Included evaluation 9 Identify, Test, and QA Search Terms

Chemical verification Web-based tool to identify and document and development of relevant search terms search terms 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") AND NOT key(human* or child* or occupat* OR infant* OR Conduct literature homind* OR woman OR women OR patient* OR OSHA OR

Identification chromatograph* OR Spectrometr* OR pediatric*) searches

Identify and acquire Couple hour process potentially applicable studies Screening

Review literature for applicability to ECOTOX Eligibility Chemical terms Data extraction automatically formatted for abstracting databases Study quality Included evaluation 10 ECOTOX Literature Searches

Search Engines Chemical verification Chemical specific searches 1. Science Direct and development of (using terms from chemical verification step) 2. AGRICOLA search terms OR 3. TOXNET 4. ProQuest ESPM Monthly electronic searches 5. ProQuest Dissertation of 11 highly relevant journals Abstracts 6. Web of Science/ Conduct literature Current Contents Identification searches

Identify and acquire potentially applicable studies Screening

Review literature for applicability to ECOTOX In 2019: 159,727 Eligibility references were manually skimmed Data extraction for applicability Study quality

Included Collate data and remove duplicates evaluation 11 ECOTOX Literature Searches Excel-based tool Search Engines Chemical verification 1. Science Direct and development of (Abstract-Sifter Plus) to 2. AGRICOLA search terms search multiple sources 3. TOXNET 4. ProQuest ESPM 5. ProQuest Dissertation Abstracts 6. Web of Science/ Conduct literature Current Contents Identification searches

Identify and acquire potentially applicable studies Screening

Review literature for applicability to ECOTOX References can be Eligibility ‘sifted’, reviewed, or exported as .ris Data extraction

Study quality

Included Collate data and remove duplicates evaluation 12 Skimming for Applicability:

Chemical verification Title and Abstract and development of search terms

Conduct literature Identification searches

Skim titles and Identify and acquire abstracts, use exclusion potentially applicable criteria to eliminate studies Screening non-applicable

Review literature for applicability to ECOTOX Eligibility

Data extraction Send applicable reference Study quality list for acquisition Included evaluation 13 Skimming for Applicability:

Chemical verification Title and Abstract and development of Partnering with NTP/SCIOME to develop language search terms learning tool for skimming/prioritizing abstracts

Conduct literature Identification searches

Skim titles and Identify and acquire abstracts, use exclusion potentially applicable criteria to eliminate studies Screening non-applicable

Review literature for applicability to ECOTOX Eligibility

Data extraction Send applicable reference Study quality list for acquisition Included evaluation 14 Skimming for Applicability:

Chemical verification Full text and development of search terms

Conduct literature Identification searches

Identify and acquire potentially applicable studies Screening

Review literature for applicability to Moves on to be curated ECOTOX

Eligibility into ECOTOX.

Data extraction Dec. 2018 – Dec. 2019 1,468 References were Study quality added to the public website Included evaluation 15 Skimming for Applicability:

Chemical verification Full text and development of Exploring options for data mining search terms and extraction of information from a variety of sources

Conduct literature Identification searches

Identify and acquire potentially applicable studies Screening

Review literature for applicability to Moves on to be curated ECOTOX

Eligibility into ECOTOX.

Data extraction Dec. 2018 – Dec. 2019 1,468 References were Study quality added to the public website Included evaluation 16 ECOTOX Applicability Criteria

• Paper must meet these criteria • Single chemical exposure Review literature for • Ecologically-relevant species applicability to • Must be able to verify CAS registry numbers ECOTOX • Must be able to verify taxonomic information for test species • Exposure to live organism, viable tissue or cells • Report concurrent exposure concentration, dose or application rate • Report duration of exposure • Must have a control treatment • Primary source of the data • Study must be a full article in English • The following studies are excluded • Air pollution studies related to CO2 and ozone • Studies on humans, monkeys, bacteria, viruses and yeast • Review and summary articles • Terrestrial studies with an inhalation route of exposure • Non-English publications and abstracts

17 ECOTOX Applicability Criteria

All Excluded and Non-Applicable studies are Tagged with the reason for rejection

• Abstract – Published as an abstract • No Conc – the authors report a response in • Bacteria – only test organism is a Bacteria an organism but do not provide • CAS # Unavailable – could not verify/locate conc/dose/app rate chemical CAS Registry number • No Duration – duration of exposure is not • Chemical method – description of chemical presented analysis procedures • No Effect – paper does not report observed • Fate – only report chemical distribution in responses adverse of otherwise media • No Toxicant (ozone, CO2) • Human Health – data on human subjects of • Non-English surrogate animal subjects for human health • Nutrient – in situ chemical tested as nutrient risk assessment • PUBL AS – duplicate data published • Incident – reports death of animal by poison, elsewhere but does not provide concentration/duration • Retracted – paper retracted by Journal of exposure • Review – primary data published elsewhere • Method – paper only reports methods for conducting a toxicity test or other aspect of • Sediment – only sediment concentration an experiment presented • Mixture – paper reports results from mixture • Survey – chemical measured in organism, of chemicals; no single chemical exposure but lack quantification of exposure results (dose/duration) • Modeling – results of the development of a • Virus – virus is only test organism model; no primary data available • Yeast – yeast is only test organism 18 ECOTOX Data Extraction

Chemical verification and development of search terms

Conduct literature Identification searches

Identify and acquire potentially applicable studies Screening

Review literature for applicability to ECOTOX Eligibility

Data extraction

Study quality Included evaluation 19 ECOTOX Data Extraction

Chemical verification and development of search terms

Conduct literature • Automated data extraction Identification searches

Identify and acquire https://www.knime.com potentially applicable studies Screening

Review literature for applicability to ECOTOX Eligibility

Data extraction

Study quality Included evaluation 20 ECOTOX Data Extraction

Chemical verification • Automated data extraction and development of search terms

Conduct literature https://www.knime.com Identification searches • Standardized unique identifiers - Chemicals: CASRN, DTXSID - Species: USGS IT IS taxonomic serial number, NCBI Taxid - Genes: NCBI Gene ID Identify and acquire - Proteins: UniProt ID, NCBI protein accession(s) potentially applicable studies • Development of ontologies for ecotoxicology Screening

Review literature for applicability to ECOTOX Eligibility

Data extraction https://www.ontotext.com

Study quality • Linking effects to biological pathways Included evaluation 21 Study Quality Evaluation

Chemical verification • Many fields in ECOTOX can inform study evaluation and development of Category Select study evaluation questions with relevant ECOTOX field(s) search terms Chemical • Is test substance identified? Required for inclusion in ECOTOX inclusion • Is the purity of test substance reported? Chemical Purity Conduct literature • Were chemical concentrations verified? Chemical Analysis (e.g.,

Identification nominal versus measured concentrations) searches Species • Is the species given? Verifiable species (Scientific Name, etc.) required for inclusion in ECOTOX • Are the organisms well described? Organism Source, Lifestage, Age, Gender, Initial and Final Weight Identify and acquire Test • Are appropriate controls performed? A control is required for potentially applicable Conditions inclusion in ECOTOX, type described in Control studies Screening • Is a guideline method (e.g., OECD) used? Test Method • Are the experimental conditions appropriate and acceptable for Review literature for the test substance and organism? Test Method, Media Type, applicability to Test Location, Experimental Design, Physical and Chemical ECOTOX Soil and Water Parameters (e.g., pH, Temperature, Dissolved Eligibility Oxygen) Test • Are the reported effects and endpoints appropriate for the Data extraction Results purpose, test substance and organism? Effect Measurement, Endpoint Study quality • Is the response/effect statistically significant? Statistical Included evaluation Significance, Significance Level 22 Study Quality Evaluation

Chemical verification • Working towards a unified study quality evaluation method and development of st search terms Pilot with 1 10 Priority TSCA Chemicals

Conduct literature Identification searches Initial Office of Water DER Templates

Critical Domains Identify and acquire o Test Substance potentially applicable o Test Design studies o Exposure Screening Characteristics Test Organism Review literature for o Outcome Assessment Test Substance Study applicability to o o o Confounding/variable Exposure Pathway Parameters ECOTOX o o Eligibility Control o Protocols o Test o Data Presentation Followed Conditions Data extraction and Analysis o Study Design o Dose/response and Methods data Study quality o Test Organism o Statistical Included evaluation Verification 23 Overview

• Background and History for ECOTOX Knowledgebase • Modernizing the ECOTOX Pipeline (C. Elonen, SOT 2020) • Mapping ecological toxicity of PFAS with ECOTOX Protocols (J. Olker, SOT 2020)

24 Background & Objectives

• Persistence and wide distribution of some PFAS in the environment – Detection of PFAS across the world in water and other media – Detection in tissue samples of invertebrates, fish, amphibians, birds, marine mammals, terrestrial mammals • Potential to bioaccumulate • Effects on ecological species • Ecological toxicity information needed to inform risk assessment and management • Sensitive and susceptible species • Bioaccumulation Across • Benchmarks and thresholds for range of ecological toxicity PFAS

Houde et al. 2006 Environ. Sci. Technol. 40:3463-3473 25 McCarthy et al. 2017 Curr. Pollution Rep. 3:289-301 Background & Objectives

• Persistence and wide distribution of some PFAS in the environment • Potential to bioaccumulate • Effects on ecological species • Ecological toxicity information needed to inform risk assessment and management

Objectives o Identify and describe available empirical evidence for ecological effects of PFAS o Identify potential ecological toxicity pathways

26 27 28 Chemical verification Chemical-based Search Terms: and development of • Chemical names and CASRNs search terms • Generic PFAS terms

Literature search: Use chemical-specific search terms to query multiple literature search engines Citations ProQuest/ Science ToxNet Dissertation Agricola Web of Already in Conduct literature from: CSA Direct Abstracts Science Unify* Identification searches n = 541 27,081 14,924 643 15,408 43,462 2,961 *Internal USEPA ECOTOX database

~145,000 citations downloaded Initial removal of duplicates Identify and acquire Not applicable (excluded): potentially applicable Title and Abstract Screening n = 15,424 references studies Chem Methods: 6,958 No Toxicant: 322 Screening n = 16,091 references Human Health: 3,755 Duplicate: 181 False Hit: 1,915 Review: 67 Fate: 670 Mixture: 15 Survey: 379 Other: 151 Review literature for Bacteria: 280 applicability to For Review (Full Text Screening) n = 565 references No PFAS in reference: ECOTOX n = 85 references Eligibility

Did not meet acceptability criteria (excluded): Extract data into Data Extracted from n = 73 references ECOTOX Acceptable Papers

Included Knowledgebase n = 462 references: 13,496 total records Awaiting Review and Data Extraction n = ~30 references 192 PFAS references Data Records for PFAS already included in n = 654 references: 18,140 total records ECOTOX 29 654 Publications, 112 PFAS with Ecological Toxicity Data

Box size represents # references that include relevant and acceptable ecological toxicity data

30 ‘Family tree’ Sub-classes of PFASs Literature Search Terms Perfluoroalkyl carboxylic acids • 322 chemical names with of PFASs (PFCAs) associated CASRNs • General PFAS search Perfluoroalkane sulfonic acids (PFSAs) terms (e.g., Dodecafluoro, Fluorotelomer, Nonafluoro, Perfluoroalkyl Pentafluoropropanoic, Perfluorobutanesulfon, Perfluoroalkyl phosphonic and Perfluoroheptanoate, acids (PFAAs) phosphinic acids Perfluorohexanoate, Perfluoropentyl) (PFPAs and PFPiAs) PFASs Per- and poly- fluoroether carboxylic acids (PFECAs) Per- and poly- fluoroether sulfonic acids (PFESAs) PASF-based substances PFAA precursors Fluorotelomer-based substances Fluoropolymers (FPs) Other PFASs Perfluoropolyethers (PFPEs)

OECD 2018 ENV/JM/MONO(2018)7 Wang et al. 2017 Environ. Sci. Technol. 51:2508-2518 31 ‘Family tree’ Sub-classes of PFASs Literature Search Terms Perfluoroalkyl carboxylic acids • 322 chemical names with of PFASs (PFCAs) associated CASRNs • General PFAS search Perfluoroalkane sulfonic acids (PFSAs) terms (e.g., Dodecafluoro, Fluorotelomer, Nonafluoro, Perfluoroalkyl Pentafluoropropanoic, Perfluorobutanesulfon, Perfluoroalkyl phosphonic and Perfluoroheptanoate, acids (PFAAs) phosphinic acids Perfluorohexanoate, Perfluoropentyl) (PFPAs and PFPiAs) PFASs Per- and poly- fluoroether carboxylic acids (PFECAs) Per- and poly- fluoroether sulfonic acids (PFESAs) PASF-based substances PFAA precursors Fluorotelomer-based substances Fluoropolymers (FPs) Other PFASs Perfluoropolyethers (PFPEs)

OECD 2018 ENV/JM/MONO(2018)7 Wang et al. 2017 Environ. Sci. Technol. 51:2508-2518 32 ‘Family tree’ Sub-classes of PFASs Examples, with CASRN # Refs Perfluoroalkyl PFOA (335-67-1) 128 carboxylic acids PFNA (375-95-1) 34 of PFASs (PFCAs) PFODA (16517-11-6) 1 K-PFOS (2795-39-3) 141 Perfluoroalkane PFHxS (108427-53-8) 5 sulfonic acids (PFSAs) NH4-PFDS (67906-42-7) 1 Perfluoroalkyl Perfluoroalkyl phosphonic and C8-PFPA (40143-78-0) 1 acids (PFAAs) phosphinic acids (PFPAs and PFPiAs) PFASs Per- and poly- fluoroether carboxylic EC236-236-8 (13252-13-6) 3 acids (PFECAs) Per- and poly- fluoroether sulfonic F-53B (73606-19-6) 11 acids (PFESAs) PASF-based Sulfluramid (4151-50-2) 41 substances FC807 (30381-98-7) 2 PFAA precursors 10:2 FTCA (53826-13-4) 5 Fluorotelomer-based 4:2 FTOH (2043-47-2) 3 substances 6:2 FTI (2043-57-4) 1 Fluoropolymers (FPs) Other PFASs Perfluoropolyethers (PFPEs)

OECD 2018 ENV/JM/MONO(2018)7 Wang et al. 2017 Environ. Sci. Technol. 51:2508-2518 33 PFAS References in ECOTOX Knowledgebase

~100 PFAS (internal list)

~200 PFAS (internal list, Draft Cross-Agency List)

~50 PFAS (Added ToxCast Set 1)

322 PFAS (Added EPA Research List, ToxCast Set 2) 23 PFAS (ToxCast replacement chems) 24 Priority PFAS 34 PFAS Data for 440 Biological Species

Midge (Chironomus riparius): 464 records Waterflea (Daphnia magna): 720 records Atlantic Salmon (Salmo salar): 800 records Zebrafish (Danio rerio): 3,264 records

35 35 Diversity in Types of Effects

PFAS records for Fish

36 Ongoing Literature Search, Review, Data Extraction

Updated list of >300 unique CASRNs and associated chemical names

Conduct literature searches

Quarterly data releases to ECOTOX Identify and acquire potentially applicable studies

Review literature for applicability to ECOTOX

Extract data and encode into ECOTOX Knowledgebase

37 Data Inventory  Summary/Synthesis 440 Biological Species 112 PFAS Measurements of 1,126 Effects

38 Data Inventory  Summary/Synthesis 440 Biological Species 112 PFAS Measurements of 1,126 Effects

Toxicant Cellular Responses Tissue/Organ Responses Organism Responses PFOS T4 & T3 ↓ Heart rate Δ Mortality ↑ (1763-23-1) Estrogen and 17-β Estradiol Δ Swim bladder inflation Δ Length & Weight ↓ or ↑ or Δ K-PFOS Vitellogenin Δ Organ:Body weight Δ Behavior (swimming, distance (2795-39-3) Acetylcholinesterase Δ Vacuolization (Liver) Δ moved) Δ Cholesterol & Lipids Δ Accumulation: Residue, Uptake Abnormal development ↑ Δ in expression of: ↑ Sperm cell counts ↓ PPAR-mediated genes (multiple) Thyroid-relevant genes (multiple) … 39 Data Inventory  Summary/Synthesis 440 Biological Species 112 PFAS Measurements of 1,126 Effects

Toxicant Cellular Responses Tissue/Organ Responses Organism Responses PFOS T4 & T3 ↓ Heart rate Δ Mortality ↑ (1763-23-1) Estrogen and 17-β Estradiol Δ Swim bladder inflation Δ Length & Weight ↓ or ↑ or Δ K-PFOS Vitellogenin Δ Organ:Body weight Δ Behavior (swimming, distance (2795-39-3) Acetylcholinesterase Δ Vacuolization (Liver) Δ moved) Δ Cholesterol & Lipids Δ Accumulation: Residue, Abnormal development ↑ Δ in expression of: Uptake ↑ Sperm cell counts ↓ PPAR-mediated genes (multiple) Thyroid-relevant genes (multiple) … 40 Identify Potential Toxicity Pathways Ontology-based semantic analysis - Bridge the gap between the molecular/non-molecular phenotypes - Lead to a better understanding of the underlying MOAs - Allow comparisons across chemicals, both within and across species

ECOTOX PFOS: Data - decreased thyroid cell extraction area - Decreased T4 - Altered expression of 7 thyroid-related genes Assemble Chemical-Species Compare to in larval zebrafish Phenotypic Profiles Phenotypic Profiles PFOS on zebrafish - Ref#178023 Genes Annotate and PFOS on zebrafish - convert to Ref#175649 Pathways ontology PFOS on zebrafish - Ref#175223 classes PFOS on zebrafish - Ref#175685 Diseases …

Wang et al. 2019 Toxicology 412:89-100 41 Identify Potential Toxicity Pathways

• 40% of the effect measurements are biochemical or genetic effects Zebrafish (Danio rerio) PFAS references include: – 252 genes measured for changes in expression – 49 biochemical measurements (e.g., proteins, enzymes, hormones)

73 Zebrafish Pathways Investigated Map to Entrez Identify Carbohydrate metabolism (3): Glycolysis/Gluconeogensis; Starch and sucrose metabolism GeneID Pathways Lipid metabolism (5): Fatty acid elongation and degradation; Steroid hormone biosynthesis Energy metabolism (1): Oxidative phosphorylation Immune system (7): Toll-like receptor signaling pathway; NOD-like receptor signaling pathway Endocrine system (6): PPAR signaling pathway; Insulin signaling pathway; Progesterone-mediated oocyte maturation Circulatory system (2): Adrenergic signaling in cardiomyocytes; Vascular smooth muscle contraction Endocrine and metabolic disease (1): AGE-RAGE signaling pathway in diabetic complications 42 Identify Potential Toxicity Pathways

73 Zebrafish Pathways Investigated Map to Entrez Identify Carbohydrate metabolism (3): Glycolysis/Gluconeogensis; Starch and sucrose metabolism GeneID Pathways Lipid metabolism (5): Fatty acid elongation and degradation; Steroid hormone biosynthesis Energy metabolism (1): Oxidative phosphorylation Immune system (7): Toll-like receptor signaling pathway; NOD-like receptor signaling pathway Endocrine system (6): PPAR signaling pathway; Insulin signaling Genes with pathway; Progesterone-mediated oocyte maturation Circulatory system (2): Adrenergic signaling in cardiomyocytes; sig. change in Vascular smooth muscle contraction transcription Endocrine and metabolic disease (1): AGE-RAGE signaling pathway in diabetic complications 43 Summary

• Extent and distribution of literature of ecological toxicity of PFAS - Curated toxicity data for multiple applications - Identification of data gaps • Literature identified for other areas of PFAS research • Ontology-based semantic analysis could advance synthesis and interpretation

• Limitations: - Mixtures currently not included - Observational and (most) field data not represented here - Limited gene and pathway information for many ecological species

44 Thank you! Questions?

Jennifer Olker, Postdoctoral Researcher [email protected]

Colleen Elonen, Jennifer Olker & Dale Hoff US EPA Office of Research and Development Center for Computational Toxicology and Exposure Great Lakes Toxicology and Ecology Division Duluth, MN http://cfpub.epa.gov/ecotox

ECOTOX Support: 218-529-5225 [email protected]

45 EXTRA SLIDES

46 Terms for Literature Search

List # of Chemicals PFAS list internal to ECOTOX 69 EPA Cross-Agency List – Chem Dashboard 199 ToxCast Set 1 List of 75 Test Samples – Chem Dashboard 74 Additional chemicals found in literature from 1st search 7 April – Nov 2018 EPA• 254 Research chemical List –namesChem Dashboardwith associated CASRNs (if applicable)165 ToxCast• General Set 2PFAS List of search 75 Test terms Samples (e.g., – ChemDodecafluoro, Dashboard Fluorotelomer, 75 ToxCastNonafluoro, Replacement Pentafluoropropanoic Test Samples, Perfluorobutanesulfon – Chem Dashboard, Perfluoroheptanoate,36 26 Perfluorohexanoate, Perfluoropentyl)

47 Terms for Literature Search

List # of Chemicals PFAS list internal to ECOTOX 69 EPA Cross-Agency List – Chem Dashboard 199 ToxCast Set 1 List of 75 Test Samples – Chem Dashboard 74 Additional chemicals found in literature from 1st search 7 EPA Research List – Chem Dashboard 165 ToxCast Set 2 List of 75 Test Samples – Chem Dashboard 75 Chem Dashboard AprilToxCast – ReplacementNov 2018 Test Samples – 36 26 • 254 chemical names with associated CASRNs (if applicable) • General PFAS search terms (e.g., Dodecafluoro, Fluorotelomer, Nonafluoro, Pentafluoropropanoic, Perfluorobutanesulfon, Perfluoroheptanoate, Perfluorohexanoate, Perfluoropentyl) July – August 2019 • 322 chemical names with associated CASRNs (if applicable) • General PFAS search terms

48 Terms for Literature Search

April – Nov 2018 • 254 chemical names with associated CASRNs (if applicable) • General PFAS search terms July – August 2019 • 322 chemical names with associated CASRNs (if applicable) • General PFAS search terms October 2019 • 23 chemical names with associated CASRNs (if applicable) 49 On-going Literature Searches for PFAS Already in ECOTOX * 2018-2019 Efforts * 185 PFAS refs Lit search for Lit search for Lit search for 23 >250 PFAS >300 PFAS ‘replacement’ March 2018 April-Nov July/Aug PFAS and before 2018 June 2019 2019 Oct 2019

247 PFAS refs added to ECOTOX Future m Lit search for Literature searches: >300 PFAS comprehensive or selective Dec 2019 March 2020 June 2020 Sept 2020 ? ?

Update from Update from July/Aug 2019 Oct 2019 Update from search search June 2020 search

50 Criteria for inclusion in ECOTOX Recently developed PECO statement for ECOTOX Requirements/Inclusionary Criteria from ECOTOX SOP P Animal: Aquatic and terrestrial species (live, whole organism) of any • Ecologically-relevant species (Population) lifestage (including preconception, in utero, lactation, peripubertal, • Live, whole organisms and adult stages). Include wild mammals (e.g. Peromyscus sp.), • Organism taxonomic information verifiable against standard insects, spiders, amphibians, birds, crustaceans, fish, molluscs, taxonomic sources reptiles, worms and invertebrates. Bacteria and viruses are not • Priority species are wild (test results for terrestrial domestic included. and laboratory species are used to fill data gaps when Plants: Aquatic and terrestrial species (live), all plants including needed) algal, moss, lichen and fungi species • In vitro studies (with viable cells or tissue) flagged for possible inclusion as requested by Programs • NOT: humans, monkeys, bacteria, viruses, yeast E Relevant forms: • Verifiable Chemical Abstract Services (CAS) number (Exposure) Chemical of Concern, name and CASRN (plus synonyms, • Single chemical exposure tradenames); when requested: Metabolites, degradants, parent • Relevant to environmental exposure compound and related chemicals • Report exposure concentration, dose or application rate Animal: Any exposure to relevant forms of the chemical of concern • Report duration of exposure including via water, injection, diet, and dermal, with reported • Sediment studies must have a water concentration concentration and duration. Inhalation studies are excluded unless reported to be included this is the primary route of environmental exposure (e.g., for • NOT: Air pollution studies related to CO2 and ozone volatile compounds). Plants: Exposure to relevant forms of the chemical of concern via water or soil, with reported concentration and duration. * Studies involving exposures to mixtures will be included only if they include exposure to a relevant form for the chemical alone. * Chemical exposures for aquatic organisms where only sediment concentrations are reported from field studies are excluded (unless porewater concentration measured); laboratory-based sediment studies are retained

C A concurrent control group exposed to vehicle-only treatment • Must have a control treatment (Comparison/ and/or untreated control (control could be a baseline Control) measurement). O All biological effects (including bioaccumulation from laboratory • Biological effect measured (Outcome) studies with concurrently measured water and tissue • Effect concurrent with associated chemical exposure concentrations). • Adverse effects are priority (beneficial, nutritional effects are lower priority) Publication/ • Primary source of the data Data Format • Study must be a full article in English • NOT: Reviews or abstract only 51 ECOTOX by the numbers

March 2019 – February 2020 (Google Analytics): 17,800 page views per month 8,400 unique page views per month

Curated ecological data from ~50,000 papers, with >11,000 chemicals and >13,000 species 52