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Effective Risk Management of Endocrine Disrupting Chemicals
Vernon Manor Hotel
Cincinnati, OH
January 29-30, 2002
Final Agenda
4-Tuesday, January 29, 2002+
7:00-8:20 AM Registration
8:30 AM Welcome and Logistics
John Cicmanec, U.S. EPA, National Risk Management Research Laboratory
Risk Management Context
Moderator: Gregory Sayles, U.S. EPA, National Risk Management Research
Laboratory
8:35 AM Purpose and Goals of the Workshop
Gregory Sayles, U.S. EPA, National Risk Management Research Laboratory
8:40 AM Welcome from the National Risk Management Research Laboratory
Lee Mulkey, U.S. EPA, National Risk Management Research Laboratory
8:50 AM Risk Management Research: Improving Environmental Decisions
Hugh McKinnon, U.S. EPA, National Risk Management Research Laboratory
9:20 AM European Community Strategy for Endocrine Disruptors: Implementation to
Date
Kathryn Tierney, European Commission
9:50 AM Break
Effects of EDCs on Humans and Wildlife
Moderator: Andy Avel, U.S. EPA, National Risk Management Research Laboratory
10:10 AM Introduction to EDCs and Their Potential Effects on Humans
Ralph Cooper, U.S. EPA, National Health and Environmental Effects
Laboratory
10:50 AM 0verview of Effects and Assessment of Endocrine-Disrupting Chemicals in
Wildlife
Gary Ankley, U.S. EPA, National Health and Environmental Effects
Laboratory
-------
11:30 AM EPA's Endocrine Disrupters Screening Program: Legislation,
Implementation, and Research
Elaine Francis, U.S. EPA, National Center for Environmental Research
12:00 PM Lunch
Exposure Assessment for EDCs
Moderator: John Cicmanec, U.S. EPA, National Risk Management Research
Laboratory
1:10 PM Development of Biological Methods to Characterize Exposure of Wildlife to
EDCs
Greg Toth, U.S. EPA, National Exposure Research Laboratory
1:40 PM Development of Chemical Methods to Characterize Exposure to EDCs in the
Neuse River Basin
Myriam Medina-Vera, U.S. EPA, National Exposure Research Laboratory
2:10 PM Monitoring Endocrine Disrupting Compounds in Aquatic Ecosystems in the
United States
Steve Goodbred, U.S. Geological Survey
2:40 PM Region 5: Ongoing Endocrine Disrupter Efforts
Lawrence Zintek, U.S. EPA, Region 5
3:10PM Break
3:30 PM Residential Indoor Air and Dust Measurements of Phthalates and Other
Endocrine Disrupting Compounds
Ruthann Rudel, Silent Spring Institute
Risk Management Approaches
Moderator: Andy Avel, U.S. EPA, National Risk Management Research Laboratory
4:00 P M EPA's Risk Management Evaluation of EDCs
Gregory Sayles, U.S. EPA, National Risk Management Research Laboratory
4:40 PM Using Bioassays to Evaluate the Performance of Risk Management
Techniques
Carolyn Acheson, U.S. EPA, National Risk Management Research
Laboratory
5:10 PM Adjourn for the day
-------
^Wednesday, January 30, 2002+
8:30 AM Welcome and Logistics for Day 2
John Cicmanec, U.S. EPA, National Risk Management Research Laboratory
Drinking Water Treatment
Moderator: James Goodrich, U.S. EPA, National Risk Management Research
Laboratory
8:35 AM Use of Granular Activated Carbon and Powdered Activated Carbon for the
Removal of EDCs from Drinking Water: A User's Guide
John Cicmanec, U.S. EPA, National Risk Management Research Laboratory
9:05 AM Evaluation of Drinking Water Treatment Technologies for Removal of
Endocrine Distrupting Compounds
Kathleen Schenck, U.S. EPA, National Risk Management Research
Laboratory
9:20 AM Risk Management of Endocrine Disrupting Chemicals (EDCs) in Drinking
Water
Frederick Pontius, Pontius Water Consultants, Inc.
9:50 AM Break
Concentrated Animal Feed Operations
Moderator: Laurel Staley, U.S. EPA, National Risk Management Research Laboratory
10:10 AM Potential of Confined Animal Feed Operations (CAFOs) to Contribute
Estrogens to the Environment
Steven Hutchins, U.S. EPA, National Risk Management Research Laboratory
10:40 AM Investigations of Sorption and Transport of Hormones and Animal
Pharmaceuticals: Initial Laboratory Results
Suresh Rao, Purdue University
Linda Lee, Purdue University
11:00 AM Fate of the Endogenous Hormones 17IS-Estradiol and Testosterone in
Composted Poultry Manure and their Sorption and Mobility in Loam Soil and
Sand
Heldur Hakk, USDA, Agricultural Research Service
11:30 AM Lunch
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Wastewater Treatment
Moderator: Marc Mills, U.S. EPA, National Risk Management Research Laboratory
12:45 PM Biological Fate of Estrogenic Compounds Associated with Sewage
Treatment. A Review
Gregory Sayles, U.S. EPA, National Risk Management Research Laboratory
1:15 PM An Engineering Approach to Evaluate Estrogenic EDCs Fate During
Wastewater Treatment
Paul McCauley, U.S. EPA, National Risk Management Research Laboratory
1:30 PM Break
Other EDC Risk Management Challenges
Moderator: Andy Avel, U.S. EPA, National Risk Management Research Laboratory
1:45 PM Endocrine Disrupters from Combustion and Vehicular Emissions:
Identification and Source Nomination
Brian Gullett, U.S. EPA, National Risk Management Research Laboratory
2:00 PM Natural Recovery of PCB-Contaminated Sediments at the Sangamo-Weston /
Twelve Mile Creek/Lake Hartwell Superfund Site
Richard Brenner, U.S. EPA, National Risk Management Research Laboratory
James Lazorchak, U.S. EPA, National Exposure Research Laboratory
2:45 PM Program for the Identification and Replacement of Endocrine Disrupting
Chemicals
Douglas Young, U.S. EPA, National Risk Management Research Laboratory
3:00 PM Adjourn Workshop
Updated January 18, 2002
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
LIST OF ATTENDEES
Jim Abbott
OFFICE: 614-424-7781
FAX: 614-424-3667
abbottj@battelle.org
Cincinnati, Ohio
Battelle
505 King Avenue
Columbus, OH 43201
USA
Carolyn Acheson
OFFICE: 513-569-7190
FAX: 513-569-7105
acheson.carolyn@epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Bruce Alleman
OFFICE: 614-424-5715
FAX: 614-424-3667
allemanb@battelle.org
Battelle
505 King Avenue
Columbus, OH 43201
USA
Al Alwan
OFFICE: 312-353-2004
FAX: 312-886-0168
alwan.al@epa.gov
US EPA Regions
77 W. Jackson Boulevard
Chicago, IL 60604 USA
Gerald T. Ankley
OFFICE: 218-529-5147
FAX: 218-529-5003
ankley.gerald@epa.gov
US EPA
6120 Congdon Boulevard
Duluth, MN 55804 USA
Barry Austern
OFFICE: 513-569-7638
FAX: 513-569-7105
austern.barry@epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Andy Avel
OFFICE: 513-569-7951
FAX: 513-569-7680
avel.andy@epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
John Bankston
OFFICE: 412-778-3315
FAX: 412-778-3545
bankstonjr@sunocochemicals.com
Sunoco, Inc.
200 Neville Road
Neville Island, PA 15225 USA
Joe Bartoszek
OFFICE: 937-285-6357
FAX:
joe.bartoszek@epa.state.oh.us
Ohio EPA
401 E. Fifth Street
Dayton, OH 45402
USA
Thomas L. Baugh
OFFICE: 404-562-8275
FAX: 404-562-8269
baugh.thomasl@epa.gov
US EPA
61 Forsyth Street, SW
Atlanta, GA 30303
USA
Scott Bergreen
OFFICE: 740-380-5288
FAX: 740-385-6490
scott.bergreen@epa.state.oh.us
Ohio EPA
2195 Front Street
Logan, OH 43107
USA
FINAL
As of: Friday, January 31, 2002
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
LIST OF ATTENDEES
Cincinnati, Ohio
Gwyn Boehringer
OFFICE: 937-445-1463
FAX: 937-445-1461
gwyn.boehringer@ncr.com
NCR
1611 South Main Street, ORGH
Dayton, OH 45479-0001 USA
Rick Booth
OFFICE: 636-936-1554
FAX: 636-936-1535
rbooth@golder.com
Colder Associates, Inc.
1630 Heritage Landing
St. Charles, MO 63303-8492
USA
Dick Brenner
OFFICE: 513-569-7657
FAX: 513-569-7105
brenner.richard@epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Elizabeth A. Bruewer
OFFICE:
FAX:
bruewer@bgnet.bgsu.edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green, OH 43403 USA
Ed Burcham
OFFICE: 513-626-0715
FAX: 513-626-0115
burcham.pe@pg.com
P&G
11450 Grooms Road
BoxC13
Cincinnati, OH 45242-1434
USA
Sarah N. Bush
OFFICE:
FAX:
sbush@bgnet.bgsu.edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green, OH 43403 USA
Greg Buthker
OFFICE: 937-285-6445
FAX:
greg.buthker@epa.state.oh.us
Ohio EPA
401 East Fifth Street
Dayton, OH 45402
USA
Ronald Chambers
OFFICE: 513-352-2922
FAX: 513-352-2915
ronald.chambers@rcc.org
Cincinnati Health Department
1525 Elm Street
Cincinnati, OH 45210 USA
Lina Chang
OFFICE: 513-569-7442
FAX: 513-569-7609
chang.lina@epa.gov
US EPA
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Russ Christenson
OFFICE: 913-433-5225
FAX: 913-433-5125
russ.christenson.b@bayer.com
Bayer Corporation
17745S. Metcalf
Stilwell, KS 66085-9104 USA
U. Eric Chukwu
OFFICE: 703-784-4472
FAX: 703-784-3432
chukwueu@mcsc.usmc.mil
MARCORSYSCOM
Program Support Directorate
2033 Barnett Avenue, Suite 315
Quantico, VA 22134-5010 USA
FINAL
As of: Friday, January 31, 2002
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
LIST OF ATTENDEES
Cincinnati, Ohio
Michael Chung
OFFICE: 201-398-4314
FAX: 201-797-4399
mchung@pirnie.com
Malcolm Pimie, Inc.
17-17 Route 208 N.
Fair Lawn, NJ 07410
USA
John Cicmanec
OFFICE: 513-569-7481
FAX: 513-569-7585
cicmanec.john@epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
MSG75
Cincinnati, OH 45268 USA
Linda M. Clark
OFFICE: 215-299-6133
FAX: 215-299-6947
linda clark@fmc.com
FMC Corporation
1735 Market Street
Philadelphia, PA 19103 USA
Robert M. Clark
OFFICE: 513-569-7201
FAX: 513-569-7658
clark.robertm@epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Pat Cline
OFFICE: 352-336-5600
FAX: 352-336-6603
pcline@golder.com
Colder Associates
6241 N.W. 23rd Street
Suite 500
Gainesville, FL 32653 USA
Joan Colson
OFFICE: 513-569-7501
FAX: 513-569-7585
colson.joan@epa.gov
US EPA ORD
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Betsy Cooper
OFFICE: 206-263-3728
FAX: 206-684-1741
betsy.cooper@metrokc.gov
King County DNR
Wastewater Treatment Division
201 S. Jackson Street
Seattle, WA 98104 USA
Ralph Cooper
OFFICE: 919-541-4084
FAX: 919-541-5138
cooper.ralph@epa.gov
US EPA NHEERL
MD-72
Research Triangle Park, NC 27711
USA
Easter A. Coppedge
OFFICE: 919-541-7863
FAX: 919-541-3527
coppedge.easter@epa.gov
US EPA
MD-44
Research Triangle Park, NC 27711
USA
Christie Croften
OFFICE:
FAX:
ccrofte@bgnet.bgsu .edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green, OH 43403 USA
Wendy Davis-Hoover
OFFICE: 513-569-7206
FAX:
davis-hoover.wendy@epa.gov
US EPA NRMRL
5995 Center Hill Avenue
Cincinnati, OH 45224
USA
FINAL
As of: Friday, January 31, 2002
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
LIST OF ATTENDEES
Armah de la Cruz
OFFICE: 513-569-7224
FAX: 513-569-7170
delacruz.armah@epa.gov
Cincinnati, Ohio
US EPA NERL / MCEARD / MERB
26 W. Martin Luther King Drive
Cincinnati, OH 45268-1314 USA
Shawn Dempsey
OFFICE:
FAX:
shawnd@bgnet.bgsu.edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green, OH 43403 USA
Anne Donlin
OFFICE: 513-569-4801
FAX: 513-569-4800
adonlin@queencity.com
SAIC
2260 Park Avenue
Suite 402
Cincinnati, OH 45206
USA
Mike Elovitz
OFFICE: 513-569-7642
FAX:
elovitz.michael@epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Carl Enfield
OFFICE: 513-569-7489
FAX:
enfield.carl@epa.gov
US EPA
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Gloria Ferrell
OFFICE: 919-571-4057
FAX: 919-571-4041
gferrell@usgs.gov
US Geological Survey
3916 Sunset Ridge Road
Raleigh, NC 27607 USA
Dean Finney
OFFICE: 423-378-4120
FAX: 423-378-4120
dfinney@chartertn.net
RIVEN DELL Consultants
1233 Morningside Circle
Kingsport, TN 37664 USA
Nelson Fok
OFFICE: 780-413-7936
FAX: 780-482-5383
nfok@cha.ab.ca
Capital Health, Alberta
10216-124 Street
300
Edmonton, Alberta T5N4A3
CANADA
Eric Foote
OFFICE: 614-424-7939
FAX: 614-424-3667
Battelle
505 King Avenue
Columbus, OH 43201
USA
Elaine Francis
OFFICE: 202-564-6789
FAX: 202-565-2444
francis.elaine@epa.gov
US EPA NCER
1200 Pennsylvania Ave., N.W.
8701R
Washington, DC 20460 USA
Patty Gallagher
OFFICE: 513-569-7579
FAX: 513-569-7757
gallagher.patricia@epa.gov
US EPA NERL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
FINAL
As of: Friday, January 31, 2002
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
LIST OF ATTENDEES
Cincinnati, Ohio
Achal Garg
OFFICE: 513-557-7034
FAX: 513-557-7050
achal.garg@rcc.org
City of Cincinnati
1600 Gest Street
Cincinnati, OH 45204
USA
Dennis B. George
OFFICE: 931-372-3507
FAX: 931-372-6346
dgeorge@tntech.edu
Tennessee Technological University
1020 Stadium Drive
P.O. Box 5033
Cookeville, TN 38505 USA
Emma Lou George
OFFICE: 513-569-7578
FAX: 513-569-7585
george.emmalou@epa.gov
US EPA ORD / NRMRL / TTSD / TTB
26 W. Martin Luther King Drive
Cincinnati, OH 45268-0001 USA
Susan Glassmeyer
OFFICE: 513-569-7526
FAX: 513-569-7757
glassmeyer.susan@epa.gov
US EPA NERL / MCEARD/CERB
26 W. Martin Luther King Drive
MS 564
Cincinnati, OH 45268 USA
Michael Gonzalez
OFFICE: 513-569-7998
FAX: 513-569-7677
gonzalez.michael@epa.gov
US EPA
26 W. Martin Luther King Drive
MS 443
Cincinnati, OH 45268 USA
Steve Goodbred
OFFICE: 916-278-3097
FAX: 916-278-3071
goodbred@usgs.gov
USGS, Biological Resources Division
California State University
Placer Hall 6000 J. Street
Sacramento, CA 958196129 USA
Jim Goodrich
OFFICE: 513-569-7605
FAX: 513-569-7185
goodrich.james@epa.gov
US EPA WSWRD / NRMRL / ORD
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Denise A. Gordon
OFFICE: 513-569-7594
FAX: 513-569-7609
gordon.denise@epa.gov
US EPA MERB / EERD / NERL
26 W. Martin Luther King Drive
MS 642
Cincinnati, OH 45268 USA
Patricia Grace-Jarrett
OFFICE: 502-540-6145
FAX: 502-540-6365
grace@msdlouky.org
Louisville & Jefferson Co. MSD
700 W. Liberty Street
Louisville, KY 40203 USA
Brian Gullett
OFFICE: 919-541-1534
FAX: 919-541-0290
gullett.brian@epa.gov
US EPA NRMRL
86 Alexander Drive
MD-65
Research Triangle Park, NC 27711
USA
Heldur Hakk
OFFICE: 701-239-1238
FAX: 701-239-1430
hakkh@fargo.ars.usda.gov
USDA-ARS
Biosciences Research Laboratory
P.O. Box 5674 University Station
Fargo, ND 58105-5674 USA
FINAL
As of: Friday, January 31, 2002
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
LIST OF ATTENDEES
Cincinnati, Ohio
Sharon L. Harper
OFFICE: 919-541-2443
FAX: 919-541-3527
harper.sharon@epa.gov
US EPA
MD-44
Research Triangle Park, NC 27711
USA
Fred Hayden
OFFICE: 510-526-7140
FAX: 510-526-7140
haydens@earthlink.net
Hydrosource
639 Madison Street
Albany, CA 94706
USA
Beverly Head
OFFICE: 513-557-7003
FAX: 513-557-7050
beverly.head@rcc.org
MSD of Greater Cincinnati
1600 Gest Street
Cincinnati, OH 45204 USA
Cynthia Hines
OFFICE: 513-841-4453
FAX: 513-841-4486
chines@cdc.gov
NIOSH
4676 Columbia Parkway
R-14
Cincinnati, OH 45226 USA
Susan Hoertt
OFFICE: 937-384-9940x3008
FAX: 937-384-9946
sjh@haleyaldrich.com
Haley & Aldrich, Inc.
9039 Springboro Pike
Dayton, OH 45342-4418 USA
Dave Holbrook
OFFICE: 540-953-2870
FAX:
dholbrook@vt.edu
Virginia Tech
418 Durham Hall
Blacksburg, VA 24061
USA
Lindsey Holtgreven
OFFICE: 419-422-3433
FAX:
lholtgr@bsnet.bgsu.edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green , OH 43403 USA
Michael T. Homsher
OFFICE: 419-424-4818
FAX: 419-424-4822
homsher@mail.findlay.edu
University of Findlay
1000N. Main Street
Findlay, OH 45840
USA
Dan Hopkins
OFFICE: 312-886-5994
FAX: 312-886-2737
hopkins.dan@epa.gov
US EPA
77 W. Jackson Boulevard
Chicago, IL 60604 USA
Steven R. Hutchins
OFFICE: 580-436-8563
FAX: 580-436-8703
hutchins.steve@epa.gov
US EPA NRMRL/SPRD
919 Kerr Research Drive
Ada, OK 74820 USA
Joan M. Jones
OFFICE: 207-287-7879
FAX: 207-287-7826
joan.m.jones@state.me.us
Maine DEP
17 State House Station
Augusta, ME 04333-0017
USA
FINAL
As of: Friday, January 31, 2002
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
LIST OF ATTENDEES
Cincinnati, Ohio
Jim Kariya
OFFICE: 202-564-8471
FAX: 202-564-8483
kariya.jim@epa.gov
US EPA OPPTS OSCP
1200 Pennsylvania Avenue, NW (MC 7203)
Washington, DC 20460 USA
Hailu Kassa
OFFICE: 419-372-9615
FAX: 419-372-2400
hkassa@bgnet.bgsu.edu
Bowling Green State University
223 Health Center
Ridge Street
Bowling Green, OH 43403 USA
Djanette Khiari
OFFICE: 303-734-3478
FAX: 303-730-0851
dkhiari@awwarf.com
AWWA Research Foundation
6666 W. Quincy Avenue
Denver, CO 80235 USA
Eric J. Kleiner
OFFICE: 513-569-7824
FAX: 513-569-7105
kleiner.eric@epa.gov
US EPA
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Joy R. Klosterman
OFFICE: 937-229-2806
FAX: 937-229-2503
klostejr@udri.udayton.edu
University of Dayton Research Institute
Environmental Sciences & Engineering Group
300 College Park, Kettering Labs, Room 102
Dayton, OH 45469-0132 USA
Vincent J. Kramer
OFFICE: 317-337-3137
FAX: 317-337-4557
vjkramer@dowagro.com
Dow AgroSciences
9330 Zionsville Road
Indianapolis, IN 46268
USA
Fran Kremer
OFFICE: 513-569-7346
FAX: 513-569-7620
kremer.fran@epa.gov
US EPA
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Radha Krishnan
OFFICE: 513-782-4730
FAX: 513-782-4663
rkrishnan@theitgroup.com
IT Corporation
11499 Chester Road
Cincinnati, OH 45246
USA
Jasen M. Kunz
OFFICE:
FAX:
jasenk@bgnet.bgsu.edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green, OH 43403 USA
Margaret J. Kupferle
OFFICE: 513-569-7548
FAX: 513-569-7108
kupferle.margaret@epa.gov
University of Cincinnati
P.O. Box 210071
Cincinnati, OH 45221-0071
USA
Eugene Langschwager
OFFICE: 513-579-3100
FAX: 513-579-3102
elangsch@gccc.com
Greater Cincinnati Chamber of Commerce
300 Carew Tower
441 Vine Street
Cincinnati, OH 45202-2812 USA
FINAL
As of: Friday, January 31, 2002
-------
Workshop on the Effective Risk
January 29-30, 2002
George Lawton
OFFICE: 415-467-8779
FAX:
eeletter@glawton.com
Management of Endocrine Disrupting Chemicals
LIST OF ATTENDEES Cincinnati, Ohio
- •' ' ~w ^•iT'^mKnimimisaffiBSH^^^^^^^^^^^^^^^H
Endocrine/Entrogen Letter
P.O. Box 390
Brisbane, CA 94005 USA
Jim Lazorchak
OFFICE: 513-569-7076
FAX: 513-569-7609
lazorchak.jim@epamail.epa.gov
US EPA MERB / EER / NERL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Linda Lee
OFFICE: 765-494-8612
FAX: 765-496-1107
lslee@purdue.edu
Purdue University
West Lafayette, IN 47907
USA
Peter A. Lewis
OFFICE: 513-681-5950x226
FAX: 513-632-1531
lewispeter@sunchem.com
Sun Chemical Corporation
5020 Spring Grove Avenue
Cincinnati, OH 45232 USA
Matt Longnecker
OFFICE: 919-541-5118
FAX: 919-541-2511
NIEHS EB
POBox 12233, MDA3-05
Reserach Triangle Park, NC 27709
USA
Victor Magar
OFFICE: 614-424-4604
FAX: 614-424-3667
magarv@battelle.org
Battelle
505 King Avenue
Columbus, OH 43201
USA
Kathya Mahadevan
OFFICE: 513-362-2602
FAX: 513-362-2610
mahadevank@battelle.org
Battelle Memorial Institute
655 Eden Park Drive
Suite 540
Cincinnati, OH 45202 USA
Todd Martin
OFFICE: 513-569-7682
FAX:
martin.todd@epa.gov
ORISE
26 W. Martin Luther King Drive
MS-466
Cincinnati, OH 45242 USA
Robert Masters
OFFICE: 614-898-7791
FAX: 614-898-7786
rmaste@ngwa.org
National Ground Water Association
601 Demsey Road
Westerville, OH 43081 USA
Dr. Eric F. Maurer
OFFICE: 513-556-9706
FAX: 513-556-5299
eric.maurer@uc.edu
University of Cincinnati
614RieveschlHall
Cincinnati, OH 45221 USA
Paul T. McCauley
OFFICE: 513-569-7444
FAX: 513-569-7105
mccauley.paul@epa.gov
US EPA NRMRL/LRPCD/TDB
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
FINAL
As of: Friday, January 31, 2002
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
Hugh McKinnon
OFFICE: 513-569-7689
FAX: 513-569-7549
mckinnon.hugh@epa.gov
LIST OF ATTENDEES
Cincinnati, Ohio
US EPA NRMRL
26 W. Martin Luther King Drive
MS 225
Cincinnati, OH 45268 USA
Evelyn Meagher-Hartzell
OFFICE: 513-569-5868
FAX: 513-569-4800
evelyn.m.meagher-hartzell@saic.com
SAIC
2260 Park Avenue
Suite 402
Cincinnati, OH 45206
USA
Myriam Medina-Vera
OFFICE: 919-541-5016
FAX: 919-541-3527
Medina-Vera.Myriam@epa.gov
US EPA NERL / HEASD / EMMB
79 T.W. Alexander Drive (MD-44)
Research Triangle Park
Durham, NC 27711 USA
Betty Merriman
OFFICE: 513-569-7454
FAX:
merriman.betty@epa.gov
National Council on Aging
28 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Marc A. Mills
OFFICE: 513-569-7322
FAX: 513-569-7105
mills.marc@epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
MS 420
Cincinnati, OH 45268 USA
Dick Miltner
OFFICE: 513-569-7403
FAX: 513-569-7892
miltner.richard@epa.gov
US EPA
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Jam! Montgomery
OFFICE: 703-684-2470x7146
FAX: 703-299-0742
jmontgomery@werf.org
Water Environment Research Foundation
601 Wythe Street
Alexandria, VA 22314 USA
Debdas Mukerjee
OFFICE: 513-569-7572
FAX: 513-569-7475
mukerjee.debdas@epa.gov
US EPA NCEA
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Lee Mulkey
OFFICE:
FAX:
mulkey.lee@epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Ron Newhook
OFFICE: 613-957-9576
FAX: 613-954-2486
ron_newhook@hc-sc.gc.ca
Health Canada
EHC, Tunney's Pasture 0802B1
Ottowa, Ontario K1AOL2
CANADA
Nancy Nilsen
OFFICE: 513-841-4276
FAX: 513-841-4486
nbn9@cdc.gov
NIOSH
5555 Ridge Road
MSR15
Cincinnati, OH 45213
USA
FINAL
As of: Friday, January 31, 2002
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Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
LIST OF ATTENDEES
Cincinnati, Ohio
Kazuhiko Nishioka
OFFICE: 212-997-0446
FAX: 212-944-8320
kazuhiko_nishioka@jetro. go.jp
JETRO New York
12221 Avenue of the Americas
New York, NY 10020 USA
Erik Orsak
OFFICE: 702-647-5230
FAX: 702-647-5231
Erik Orsak@fws.gov
U.S. Fish and Wildlife Service
1510 N. Decatur Boulevard
Las Vegas, NV 89108 USA
Clyde Owens
OFFICE: 919-541-1133
FAX: 919-541-0554
owens.clyde@epa.gov
US EPA APTB
86 Alexander Drive
MD-65
Durham, NC 27709
USA
J. William Owens
OFFICE: 513-627-1385
FAX: 513-627-1208
owens.jw@pg.com
Procter & Gamble
11810 E. Miami River Road
Cincinnati, OH 45252 USA
Neil J. Parke
OFFICE: 317-276-7201
FAX: 317-276-1800
parke_neil J@lilly.com
Eli Lilly and Company
Lilly Corporate Center
Indianapolis, IN 46285 USA
David Paulsen
OFFICE: 702-450-4440
FAX: 702-454-7966
dpaulsen@co.clark.nv.us
Clark Co. Sanitation District
5857 E. Flamingo Road
Las Vegas, NV 89122 USA
Dan Petersen
OFFICE: 513-569-7831
FAX: 513-569-7585
US EPA
26 W. Martin Luther King Drive
Cincinnati, OH 45208 USA
Frank Pierik
OFFICE: +31 104087448
FAX: +31 104089455
pierik@mgz.egg.eur.nl
Erasmus MC Dept. of Pulic Health
P.O. Box 1738
Rotterdam, 3000 DR THE NETHERLANDS
Fred Pontius
OFFICE: 303-986-9923
FAX: 303-716-7310
fredp@pontiuswater.com
Pontius Water Consultants, Inc.
P.O. Box 150361
Lakewood, CO 80215 USA
Suresh Rao
OFFICE: 765-496-6554
FAX: 765-496-1107
pscr@purdue.edu
Purdue University
School of Civil Engineering
West Lafayette, IN 479071284
USA
Tirumuru V. Reddy
OFFICE: 513-569-7295
FAX: 513-569-7609
reddy.tirumuru@epa.gov
US EPA NERL / EERD / MERB
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
FINAL
10
As of: Friday, January 31, 2002
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002 LIST OF ATTENDEES Cincinnati, Ohio
^^^^^^^^^^^^^^^^m^^mmiimimmamMm.m^*-',''"-.^- ,* ^ » - - , - ^ * •"•,„ «&-
-------
Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
LIST OF ATTENDEES
Cincinnati, Ohio
Rachel Sell
OFFICE: 513-362-2605
FAX: 513-362-2610
niemannr@battelle.org
Battelle
Battelle's ETC2
655 Eden Park Drive
Cincinnati, OH 45202
USA
Yonggui Shan
OFFICE: 513-569-7606
FAX: 513-569-7105
shan.yonggui@epa.gov
University of Cincinnati
P.O. Box 210071
Cincinnati, OH 45221
USA
Danielle T. Shirk
OFFICE:
FAX:
shirktk@bgnet.bgsu.edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green, OH 43403 USA
Kaniz Siddiqui
OFFICE:
FAX:
kaniz.f.siddiqui@rcc.org
MSD City of Cincinnati
1600 Gest Street
Cincinnati, OH 45201 USA
Sukh Sidhu
OFFICE: 937-229-3605
FAX: 937-229-2503
sidhu@udri.udayton.edu
University of Dayton
Environmental Science & English
300 College Park, UDRI, KL102
Dayton, OH 45469-0132 USA
Gary Silverman
OFFICE: 419-372-6062
FAX: 419-372-2400
silverma@bgnet.bgsu.edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green, OH 43403 USA
Guy Simes
OFFICE: 513-579-7685
FAX: 513-569-7471
simes.guy@epamail.epa.gov
US EPA
26 W. Martin Luther King Drive
Cincinnati, OH 45368 USA
Hugh M. Smith
OFFICE: 513-681-5950x272
FAX: 513-632-1531
smithh@sunchem.com
Sun Chemical Corporation, Colors Group
5020 Spring Grove Avenue
Cincinnati, OH 45232 USA
Louise T. Snyder
OFFICE: 937-285-6475
FAX:
louise.snyder@epa.state.oh.us
Ohio EPA
401 E. Fifth Street
Dayton, OH 45402
USA
Adam Socha
OFFICE: 416-327-3912
FAX: 416-327-9091
adam.socha@ene.gov.on.ca
Ontario Ministry of the Environment
Standards Development Branch
125 Resources Road, North Wing
Etobicoke, Ontario M9P 3V6 CANADA
Thomas Speth
OFFICE: 513-569-7208
FAX: 513-569-7892
speth.thomas@epa.gov
US EPA
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
FINAL
12
As of: Friday, January 31, 2002
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Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002 LIST OF ATTENDEES Cincinnati, Ohio
Sara Spino
OFFICE:
FAX:
sspino@bgnet.bgsu.edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green, OH 43403
USA
Laurel Staley
OFFICE: 513-569-7863
FAX: 513-569-7105
staley.laurel@epamail.epa.gov
US EPA NRMRL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
T. R. Steinheimer
OFFICE: 515-294-2952
FAX: 515-294-8125
steinheimer@nstl.gov
USDA ARS
National Soil Tilth Laboratory
2150 Pammel Drive
Ames, IA 50011-4047 USA
Tammy Stoker
OFFICE: 919-541-2783
FAX: 919-541-5138
stoker.tammy@epa.gov
US EPA NHEERL/GEEB
2525 Alexander Drive
MD-72
Research Triangle Park, NC 27713
USA
Gary Strassell
OFFICE: 513-874-0714
FAX: 513-874-5061
gstrassell@shepherdcolor.com
The Shepherd Color Company
PO Box 465627
Cincinnati, OH 45246 USA
Richard C. Striebich
OFFICE: 937-229-2847
FAX: 937-229-2503
striebich@udri.udayton.edu
University of Dayton Research Institute
300 College Park
KL102
Dayton, OH 45469-0132 USA
Daniel Sullivan
OFFICE: 616-833-0394
FAX:
daniel.e.sullivan@pharmacia.com
Pharmacia Corporation
7000 Portage Road
MS 6605-88-009
Kalamazoo, Ml 49001 USA
Kathryn Tierney
OFFICE: 003-2229-68118
FAX: 003-2229-91067
kathryn.tierney@cec.eu.int
Europeon Commission
200 Rue De La Loi (B45 2/157)
Brussels, Belguium 1049 Europe
Greg Toth
OFFICE: 513-569-7242
FAX: 513-569-7609
toth.greg@epa.gov
US EPA NERL
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Marcus Tsilimos
OFFICE:
FAX:
marcust@bgnet.bgsu.edu
Bowling Green State University
Environmental Health Program
223 Health Center
Bowling Green, OH 43403 USA
Raghuraman Venkatapathy
OFFICE: 513-569-7077
FAX: 513-569-7475
venkatapathy.raghuraman@epa.gov
US EPA NCEA/ORISE
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
FINAL
13
As of: Friday, January 31, 2002
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Workshop on the Effective Risk Management of Endocrine Disrupting Chemicals
January 29-30, 2002
Karl G. Voelkel
OFFICE: 937-259-5041
FAX: 937-259-5100
kvoelkel@ljbinc.com
LIST OF ATTENDEES
Cincinnati, Ohio
LJB, Inc.
3100 Research Boulevard
Dayton, OH 45420-0246 USA
Mark A. Walton
OFFICE: 208-938-5130
FAX: 208-938-5662
mawalton@dow.com
The Dow Chemical Company
1924 S. Riverford Place
Eagle, ID 83616USA
David Warshawsky
OFFICE: 513-558-0152
FAX:
University of Cincinnati
P.O. Box 210071
Cincinnati, OH 45221-0071
USA
Martha J. M. Wells
OFFICE: 931-372-3507
FAX: 931-372-6346
mjmwells@tntech.edu
Tennessee Technological University
P.O. Box 5033
1020 Stadium Drive
Cookeville, TN 38505 USA
Dennis J. Wesolowski
OFFICE: 312-353-9084
FAX: 312-886-2591
wesolowski.dennis@epa.gov
US EPA Region 5
536 S. Clark Street
Chicago, IL 60605
USA
Elizabeth Whelan
OFFICE: 513-841-4437
FAX: 513-841-4486
EWhelan@cdc.gov
National Institute for Occupational Safety and Health
4676 Columbia Parkway
R-15
Cincinnati, OH 45226 USA
Greg Woodside
OFFICE: 714-378-3275
FAX: 714-378-3369
gwoodside@ocwd.com
Orange County Water District
10500 Ellis Avenue
Fountain Valley, CA 92626
USA
Michael Wright
OFFICE: 513-569-7922
FAX: 513-569-7475
wright.michael@epa.gov
ORISE-NCEA
140 Warner Street
#1
Cincinnati, OH 45219
USA
Douglas Young
OFFICE: 513-569-7624
FAX: 513-569-7111
young.douglas@epa.gov
US EPA ORD / NRMRL / STD / SAB
26 W. Martin Luther King Drive
Cincinnati, OH 45268 USA
Diana Zimmerman
OFFICE: 937-285-6440
FAX: 937-285-6249
diana.zimmerman@epa.state.oh.us
Ohio EPA
401 E. Fifth Street
Dayton, OH 45402
USA
Lawrence Zintek
OFFICE: 312-886-2925
FAX: 312-886-2591
zintek.lawrence@epa.gov
US EPA Regions
77 W. Jackson Boulevard
MCML-10C
Chicago, IL 60605 USA
FINAL
14
As of: Friday, January 31, 2002
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Effective Risk Management of Endocrine Disrupting
Chemicals
January 29-30, 2002
Cincinnati, Ohio
Sponsored by
U.S. Environmental Protection Agency
Office of Research and Development
National Risk Management Research Laboratory
-------
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Effective Risk Management of Endocrine Disrupting Chemicals
Vernon Manor Hotel
Cincinnati, OH
January 29-30, 2002
Final Agenda
* Tuesday, January 29, 2002s
7:00-8:20 AM Registration
8:30 AM Welcome and Logistics
John Cicmanec, U.S. EPA, National Risk Management Research Laboratory
Risk Management Context
Moderator: Gregory Sayles, U.S. EPA, National Risk Management Research
Laboratory
8:35 AM Purpose and Goals of the Workshop
Gregory Sayles, U.S. EPA, National Risk Management Research Laboratory
8:40 AM Welcome from the National Risk Management Research Laboratory
Lee Mulkey, U.S. EPA, National Risk Management Research Laboratory
8:50 AM Risk Management Research: Improving Environmental Decisions
Hugh McKinnon, U.S. EPA, National Risk Management Research Laboratory
9:20 AM European Community Strategy for Endocrine Disruptors: Implementation to
Date
Kathryn Tierney, European Commission
9:50 AM Break
Effects of EDCs on Humans and Wildlife
Moderator: Andy Avel, U.S. EPA, National Risk Management Research Laboratory
10:10 AM Introduction to EDCs and Their Potential Effects on Humans
Ralph Cooper, U.S. EPA, National Health and Environmental Effects
Laboratory
10:50 AM Overview of Effects and Assessment of Endocrine-Disrupting Chemicals in
Wildlife
Gary Ankley, U.S. EPA, National Health and Environmental Effects
Laboratory
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11:30 AM EPA's Endocrine Disrupters Screening Program: Legislation,
Implementation, and Research
Elaine Francis, U.S. EPA, National Center for Environmental Research
12:00 PM Lunch
Exposure Assessment for EDCs
Moderator: John Cicmanec, U.S. EPA, National Risk Management Research
Laboratory
1:10 PM Development of Biological Methods to Characterize Exposure of Wildlife to
EDCs
Greg Toth, U.S. EPA, National Exposure Research Laboratory
1:40 PM Development of Chemical Methods to Characterize Exposure to EDCs in the
Neuse River Basin
Myriam Medina-Vera, U.S. EPA, National Exposure Research Laboratory
2:10 PM Monitoring Endocrine Disrupting Compounds in Aquatic Ecosystems in the
United States
Steve Goodbred, U.S. Geological Survey
2:40 PM Region 5: Ongoing Endocrine Disruptor Efforts
Lawrence Zintek, U.S. EPA, Region 5
3:10PM Break
3:30 PM Residential Indoor Air and Dust Measurements of Phthalates and Other
Endocrine Disrupting Compounds
Ruthann Rudel, Silent Spring Institute
Risk Management Approaches
Moderator: Andy Avel, U.S. EPA, National Risk Management Research Laboratory
4:00 PM EPA's Risk Management Evaluation of EDCs
Gregory Sayles, U.S. EPA, National Risk Management Research Laboratory
4:40 PM Using Bioassays to Evaluate the Performance of Risk Management
Techniques
Carolyn Acheson, U.S. EPA, National Risk Management Research
Laboratory
5:10PM Adjourn for the day
-------
s Wednesday, January 30, 2002s
8:30 AM Welcome and Logistics for Day 2
John Cicmanec, U.S. EPA, National Risk Management Research Laboratory
Drinking Water Treatment
Moderator: James Goodrich, U.S. EPA, National Risk Management Research
Laboratory
8:35 AM Use of Granular Activated Carbon and Powdered Activated Carbon for the
Removal ofEDCs from Drinking Water: A User's Guide
John Cicmanec, U.S. EPA, National Risk Management Research Laboratory
9:05 AM Evaluation of Drinking Water Treatment Technologies for Removal of
Endocrine Distrupting Compounds
Kathleen Schenck, U.S. EPA, National Risk Management Research
Laboratory
9:20 AM Risk Management of Endocrine Disrupting Chemicals in Drinking Water
Frederick Pontius, Pontius Water Consultants, Inc.
9:50 AM Break
Concentrated Animal Feed Operations
Moderator: Laurel Staley, U.S. EPA, National Risk Management Research Laboratory
10:10 AM Potential of Confined Animal Feed Operations (CAFOs) to Contribute
Estrogens to the Environment
Steven Hutchins, U.S. EPA, National Risk Management Research Laboratory
10:40 AM Investigations of Sorption and Transport of Hormones and Animal
Pharmaceuticals: Initial Laboratory Results
Suresh Rao, Purdue University
Linda Lee, Purdue University
11:00 AM Fate of the Endogenous Hormones 17K-Estradiol and Testosterone in
Composted Poultry Manure and their Sorption and Mobility in Loam Soil and
Sand
Heldur Hakk, USDA, Agricultural Research Service
11:30 AM Lunch
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Wastewater Treatment
Moderator: Marc Mills, U.S. EPA, National Risk Management Research Laboratory
12:45 PM Biological Fate of Estrogenic Compounds Associated with Sewage
Treatment A Review
Gregory Sayles, U.S. EPA, National Risk Management Research Laboratory
1:15 PM An Engineering Approach to Evaluate Estrogenic EDO's Fate During
Wastewater Treatment
Paul McCauley, U.S. EPA, National Risk Management Research Laboratory
1:30 PM Break
Other EDC Risk Management Challenges
Moderator: Andy Avel, U.S. EPA, National Risk Management Research Laboratory
1:45 PM Endocrine Disruptors from Combustion and Vehicular Emissions:
Identification and Source Nomination
Brian Gullett, U.S. EPA, National Risk Management Research Laboratory
2:00 PM Natural Recovery of PCB-Contaminated Sediments at the Sangamo-Weston /
Twelve Mile Creek/Lake Hartwell Superfund Site
Richard Brenner, U.S. EPA, National Risk Management Research Laboratory
James Lazorchak, U.S. EPA, National Exposure Research Laboratory
2:45 PM Program for the Identification and Replacement of Endocrine Disrupting
Chemicals
Douglas Young, U.S. EPA, National Risk Management Research Laboratory
3:00 PM Adjourn Workshop
Updated January 18, 2002
VI
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TABLE OF CONTENTS
Risk Management Context 1
Effects of EDCs on Humans and Wildlife 9
Exposure Assessment for EDCs 17
Risk Management Approaches 31
Drinking Water Treatment 37
Other EDC Risk Management Challenges 67
Vll
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Vlll
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s Tuesday, January 29, 2002s
Risk Management Context
Moderator: Gregory Sayles, U.S. EPA, National Risk Management Research
Laboratory
8:35 AM Purpose and Goals of the Workshop
Gregory Sayles, U.S. EPA, National Risk Management Research Laboratory
8:40 AM Welcome from the National Risk Management Research Laboratory
Lee Mulkey, U.S. EPA, National Risk Management Research Laboratory
8:50 AM Risk Management Research: Improving Environmental Decisions
Hugh McKinnon, U.S. EPA, National Risk Management Research Laboratory
9:20 AM European Community Strategy for Endocrine Disrupters: Implementation to
Date
Kathryn Tierney, European Commission
9:50 AM Break
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WELCOME FROM THE NATIONAL RISK MANAGEMENT RESEARCH LABORATORY
Lee Mulkey
U.S. Environmental Protection Agency
National Risk Management Research Laboratory
26 West Martin Luther King Drive
Cincinnati, OH 45268
513-569-7689
mulkey.lee@epa.gov
THE SPEAKER'S ABSTRACT IS UNAVAILABLE, PLEASE SEE THE SPEAKER WITH
COMMENTS AND/OR QUESTIONS.
Lee A. Mulkey
Lee A. Mulkey is the Associate Director of Ecology at the U.S. EPA's National Risk Management
Research Laboratory in Cincinnati, OH. He is responsible for developing and providing science
policy guidance for EPA's research on protecting ecosystems. His professional experience has
focused on watershed management research, including: modeling, best management practice
development and testing, and risk assessment.
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RISK MANAGEMENT RESEARCH: IMPROVING ENVIRONMENTAL DECISIONS
Hugh W. McKinnon, M.D., M.P.H.
U.S. Environmental Protection Agency
National Risk Management Research Laboratory
26 West Martin Luther King, Jr. Drive
Cincinnati, OH 45268
(513) 569-7689
(513) 569-7549 fax
mckinnon.hugh@epa.gov
Over the past two decades much progress has been made in defining and implementing the disciplines and
processes of risk assessment, including research to reduce uncertainties in risk assessment. Similar definition
is needed for the risk management process, and risk management research is needed to provide this definition
and reduce uncertainties in risk management. This presentation will describe some of the needs and issues
associated with risk management research, including creation of risk management evaluations (RME) and a
protocol for developing or conducting risk management evaluations.
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Hugh W. McKinnon, M.D., M.P.H.
Dr. Hugh McKinnon is a Senior Executive Medical Officer in the U.S. Environmental Protection
Agency (EPA) and is the Associate Director for Health in EPA's National Risk Management
Research Laboratory (NRMRL) in Cincinnati, Ohio, a position he has held since May 1995. From
1989 to 1995 Dr. McKinnon was Director of EPA's Human Health Assessment Group in
Washington, DC, which produced EPA's January 1993 report on respiratory effects, including lung
cancer, associated with exposure to environmental tobacco smoke (ETS). Earlier Dr. McKinnon
served for ten years as a Medical Officer in EPA's Office of Health Research, also in Washington,
DC, and was the Acting Director of that office from 1985 to 1987. He received a Doctor of Medicine
(M.D.) Degree from the University of Virginia in 1977 and a Master of Public Health (M.P.H.)
Degree from The John Hopkins University, where he completed the residency in General Preventive
Medicine in 1989. He has also had clinical training in general surgery and in family practice. Dr.
McKinnon is a member of and serves on committees and boards of the American College of
Occupational and Environmental Medicine (ACOEM), the American Public Health Association
(APHA), and the Federal Physicians Association. He also serves as the EPA representative to the
National Cancer Advisory Board of the National Cancer Institute.
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EUROPEAN COMMUNITY STRATEGY FOR ENDOCRINE DISRUPTERS:
IMPLEMENTATION TO DATE
Kathryn Tierney
European Commission, Environment Directorate-General
200 Rue de la Loi (BUS 2/157), B-1049 Brussels, Belgium
Tel +32-2-296 8118; Fax +32-2-299 1067; kathryn.tiernev(a)cec.eu.int
In December 1999, the European Commission adopted a Communication to the Council and
European Parliament on a Community Strategy for Endocrine Disrupters (COM(1999)706). The
strategy addresses the key requirements of further research, international co-operation,
communication to the public, and appropriate policy action. Recommendations are made for short-,
medium- and long-term actions.
In March 2000, the Environment Council of Ministers adopted Conclusions on the Commission
Communication in which it stressed the precautionary principle, the need to develop quick and
effective risk management strategies and the need for consistency with the overall EU chemical
policy.
In October 2000, the European Parliament adopted a Resolution on endocrine disrupters,
emphasising the application of the precautionary principle and calling on the European Commission
to identify substances for immediate action.
A key short-term action of the Community Strategy is the establishment of a priority list of
substances for further evaluation of their role in endocrine disruption. During 2000, a candidate list
of 553 man-made substances and 9 synthetic/natural hormones was identified and a priority list of
actions has been developed in order to further evaluate the role of these substances in endocrine
disruption.
In June 2000, the European Commission organised a European Workshop on Endocrine Disrupters in
Sweden, which focused on information exchange and international co-operation, research and
development, test methods/testing strategy and establishment of monitoring programmes.
The Commission and Member States continue to participate in the OECD Endocrine Disrupter
Testing and Assessment Task Force, which was set up in 1998 with the goal of developing agreed
test methods for endocrine disrupters.
Under the 5th Community Framework Programme for R&D (1999-2002), research into endocrine
disruption has been prioritised. In May 2001, a dedicated call for research proposals on the health
and environmental implications of endocrine disrupters was published with a budgetary envelope of
20 MEURO.
Finally, regarding legislative action, the revision of the General Product Safety Directive will lead,
inter-alia, to a simplification of conditions and procedures for urgent measures at Community level.
In addition, the issue of endocrine disrupters is addressed specifically in the context of new and
existing legislation in the field of water policy and in the White Paper on a strategy for a future
chemicals policy.
REFERENCE
Communication from the Commission to the Council and European Parliament on the
implementation of the Community Strategy for Endocrine Disrupters (COM(2001)262).
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Kathryn Tierney
Kathryn Tierney is a Principal Administrator with the European Commission. She has a Bachelor's
Degree in Industrial Engineering and a Master's Degree in Engineering Science from the National
University of Ireland. She joined the European Commission in 1991 and first worked on the
establishment of an international research programme involving Australia, Canada, Japan, the USA
and the EU/EFTA countries. In 1997 she joined the Environment Directorate-General and has been
working on the topic of endocrine disruption from that time. Before joining the Commission, Ms.
Tierney worked for Digital Equipment Corporation in Ireland and the US and for the Irish Science
and Technology Agency.
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s Tuesday, January 29, 2002s
Effects of EDCs on Humans and Wildlife
Moderator: Andy Avel, U.S. EPA, National Risk Management Research Laboratory
10:10 AM Introduction to EDCs and Their Potential Effects on Humans
Ralph Cooper, U.S. EPA, National Health and Environmental Effects
Laboratory
10:50 AM Overview of Effects and Assessment of Endocrine-Disrupting Chemicals in
Wildlife
Gary Ankley, U.S. EPA, National Health and Environmental Effects
Laboratory
11:30 AM EPA's Endocrine Disrupters Screening Program: Legislation,
Implementation, and Research
Elaine Francis, U.S. EPA, National Center for Environmental Research
12:00 PM Lunch
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10
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INTRODUCTION TO EDCs AND THEIR POTENTIAL EFFECTS ON HUMANS
Ralph L. Cooper, Chief
Endocrinology Branch, Reproductive Toxicology Division, National Health and Environmental
Effects Research Laboratory, U.S. EPA
Research Triangle Park, NC 27711
(919) 541-4084, (919)541-5138 fax
cooper.ralph@epa.gov
The endocrine system provides important regulatory support to the homeostatic mechanisms
involved in a variety of physiological functions. These precisely timed events require appropriate
signaling between the various organs which includes the synthesis, release, transport identification
and clearance of individual hormones associated with such complex processes as puberty,
reproduction, adaptation to stress, normal metabolic function, and behavior. As such, the endocrine
system provides a number of target sites that may be susceptible to disruption by environmental
agents. The purpose of this discussion will be to provide a brief overview of the hormonal control of
reproductive and thyroid function including, a description of the organs and hormones involved.
This will be followed by a summary of the cellular mechanisms of primary concern to the issue of
endocrine disruption such as the enzymes involved in the synthesis of hormones, nuclear and
membrane receptors, and the processes involved in metabolism (clearance). The issue of endocrine
disrupters will then be discussed focusing on those reports that have raised concern that
environmental endocrine disrupters may be responsible for impaired human reproductive health.
This discussion will be followed by a review of the ongoing efforts of EPAs Endocrine Disrupter
Screening Program as it relates to evaluating chemicals for their potential endocrine disrupting
effects on mammalian reproduction, reproductive development and thyroid function.
This abstract does not represent EPA policy.
11
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Ralph Cooper
Dr. Ralph Cooper is the Chief of the Endocrinology Branch at the U.S. EPA National Health and
Environmental Effects Laboratory (NHEERL) Reproductive Toxicology Division (RTD) (1995-
present). Dr. Cooper was the chief of Endocrinology/Gerontology Section at NHEERL RTD from
1984-1995. He has a Ph.D. in Psychobiology from Rutgers University and a NIH Post Doctorate
from Duke University's Neuroscience Program. Dr. Cooper is currently the Chair of the NHEERL
Endocrine Disrupter Research Implementation Committee (2000-present). Dr. Cooper co-chaired the
RTD/Program Office and Region Workshop on Emerging Issues in Reproductive Toxicology in June
2001. Dr. Cooper is also a member of Sigma Xi, the North Carolina Chapter of the Society for
Neuroscience, and the Endocrine Society. He is the recipient of numerous awards including: 1) Best
Paper Published in Reproductive & Developmental Toxicology Specialty Section, Toxicological
Sciences (2001); 2) the EDSP team award for Exceptional/Outstanding ORD Technical Assistance to
the Regions or Program Offices (2001); 3) the Office of Pesticides Programs Health Effects Division
Team Award for work related to the chlorotriazines (2000); 4) Best Paper Published in Reproductive
& Developmental Toxicology Specialty Section, Toxicological Sciences (1999); and 5) a Bronze
Medal ORD/RAF Workgroup on Environmental Endocrine Disruption: An Effects Assessment and
Analysis Document (1998).
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OVERVIEW OF EFFECTS AND ASSESSMENT OF ENDOCRINE-DISRUPTING
CHEMICALS IN WILDLIFE
Gary Ankley
U.S. Environmental Protection Agency
Environmental Effects Research Laboratory
6201Congdon Blvd.
Duluth, MN 55804
(218) 529-5147
ankley. gerald@epa. gov
THE SPEAKER'S ABSTRACT IS UNAVAILABLE, PLEASE SEE THE SPEAKER WITH
COMMENTS AND/OR QUESTIONS.
Gerald T. (Gary) Ankley
Dr. Ankley is the Chief of the U.S. EPA Mid-Continent Ecology Division, Toxic Effects
Characterization Branch. He is also an adjunct professor in the Department of Biology at Michigan
Technological University and the Department of Fisheries and Wildlife at the University of
Minnesota. Dr. Ankley is the Aquatic Toxicology Editor for Environmental Toxicology and
Chemistry and is a member of the Editorial Board for Chemosphere. He has a Ph.D. and MS in
Forest Resources (Aquatic Toxicology) from the University of Georgia. Dr. Ankley has received
numerous awards, including: fourteen EPA awards in recognition of different aspects of leadership,
scientific planning, and contributions to specific research projects; his manuscript was nominated for
best paper in the 2000 volume of North America Journal of Aquaculture; and the Scientific and
Technology Achievement Award (STAA) for journal articles on development and application of
chronic sediment test with Chironomus tentans.
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14
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EPA's ENDOCRINE DISRUPTORS SCREENING PROGRAM: LEGISLATION,
IMPLEMENTATION, AND RESEARCH
Elaine Z. Francis
U.S. Environmental Protection Agency
Office of Research and Development/National Center for Environmental Research
1200 Pennsylvania Avenue, NW (8701R)
Washington, DC 20460
(202) 564-6789
(202) 565-2444 fax
francis.elaine@epa.gov
In 1996, the Food Quality Protection Act (FQPA) and the Safe Drinking Water Act Amendments
(SDWAA) were enacted and specifically require the testing of pesticides and other chemicals found
in or on food or in drinking water sources to determine their "estrogenic or other endocrine effects in
humans." These laws required EPA to develop a screening program by August 1998, to implement
the program by August 1999, and to report to Congress on the program's progress by August 2000.
The Agency sought advice on how to design such a program by convening the Endocrine Disrupters
Screening and Testing Advisory Committee (EDSTAC), a 39-member panel representing broad
constituencies. EDSTAC issued a final report in August 1998 that included 71 specific
recommendations (www.epa.gov/scipoly/oscpendohistory/). Among the key recommendations were
that EPA's endocrine disrupters screening program (EDSP) should be expanded beyond the
provisions of FQPA, taking into consideration other EPA mandates and, thus, should: 1) address
both human and ecological (wildlife) effects, 2) examine effects to not only estrogen but also
androgen and thyroid processes, and 3) evaluate endocrine disrupting properties of chemicals and
common mixtures. EDSTAC recommended a tiered approach that included: initial sorting, priority
setting, Tier 1 screening and Tier 2 testing. They recommended a battery of eight in vitro and in vivo
assays for Tier 1 screening with four additional assays as alternatives. Tier 2 testing
recommendations included multi-generation studies conducted in mammalian, avian, fish,
invertebrate, and amphibian species. EPA's Office of Prevention, Pesticides, and Toxic Substances
has developed a program to implement the legislative mandates taking into consideration the
EDSTAC recommendations. They have held a series of public meetings and workshops on various
aspects of the program. The report to Congress issued August 2000 describes EPA's implementation
of the science-driven EDSP. Implementation activities include completing the Endocrine Disrupter
Priority Setting Database and the compartment-based approach, that the Agency will use to establish
priorities for screening chemicals at a later stage of implementation, and ensuring that the Tier 1
screens and Tier 2 tests are validated as required by statute. Research conducted by EPA's Office of
Research and Development (ORD) is addressing the scientific questions that have arisen as a result
of the FQPA and SDWAA and is leading to the development of protocols that will undergo
validation for the EDSP. The screens will be completed by 2003 and the tests by 2005. In addition,
ORD's long-term research program on ED focuses on the most critical uncertainties in determining
whether humans and wildlife are being impacted by levels of ED in the environment, in identifying
the sources of those exposures, developing approaches to integrate information into risk assessments,
and developing risk management approaches to reduce/prevent exposures.
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Elaine Z. Francis
Dr. Elaine Francis has been with EPA for 21 years - first as a developmental/ reproductive
toxicologist with the Office of Toxic Substances and, since 1988, with the Office of Research and
Development. She spent 1991 as a legislative fellow to Senator Joseph Lieberman working on
pesticides, lead, and children's issues. From 1995 to 2000 she was the Pesticides, Toxics, and
Multimedia Staff Director and coordinated the planning for many of ORD's research programs
including those for toxics and pesticides, human health, ecological risk assessment, pollution
prevention, and endocrine disrupters. In January 2000, Elaine became the National Program Director
for EPA's endocrine disrupters research program. She received her doctorate in Anatomy from
Thomas Jefferson University, where the focus of her training and research was on normal and
abnormal human development and reproduction.
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s Tuesday, January 29, 2002s
Exposure Assessment for EDCs
Moderator: John Cicmanec, U.S. EPA, National Risk Management Research Laboratory
1:10 PM Development of Biological Methods to Characterize Exposure of Wildlife to
EDCs
Greg Toth, U.S. EPA, National Exposure Research Laboratory
1:40 PM Development of Chemical Methods to Characterize Exposure to EDCs in the
Neuse River Basin
Myriam Medina-Vera, U.S. EPA, National Exposure Research Laboratory
2:10 PM Monitoring Endocrine Disrupting Compounds in Aquatic Ecosystems in the
United States
Steve Goodbred, U.S. Geological Survey
2:40 PM Region 5: Ongoing Endocrine Disruptor Efforts
Lawrence Zintek, U.S. EPA, Region 5
3:10PM Break
3:30 PM Residential Indoor Air and Dust Measurements of Phthalates and Other
Endocrine Disrupting Compounds
Ruthann Rudel, Silent Spring Institute
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18
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DEVELOPMENT OF BIOLOGICAL METHODS TO CHARACTERIZE EXPOSURE OF
WILDLIFE TO EDCs
Greg Toth
U.S. Environmental Protection Agency
NERL EERD MER
26 W. Martin Luther King, Jr. Drive
Cincinnati, OH 45268
(513) 569-7242
(513) 569-7609 fax
toth.greg@epa.gov
Biological indicators of exposure are being developed to monitor surface and ground water samples
for the presence of extrogenic activity. Changes in vitellogenin gene expression in the livers of adult
male fathead minnows (Pimephales promelas) have been studied in controlled laboratory, mesocosm
and field studies. In the laboratory, following static exposure of fatheads to 17"-ethinyl estradiol for
24 hours, vitellogenin gene expression was detected at 2 ng/1. Expression has also been detected in
the gills of adult fatheads as well as in 24 hour whole larva. This indicator method has been used in
surrogate ecosystems (mesocosms) to study the attenuation of synthetic estrogens across differing
nutrient status. Field study includes the monitoring of risk management practices (e.g., sediment
capping) to evaluate their effectiveness by using similar molecular assays - changes in fathead
minnow liver cytochrome P450IA1 levels. Current biological indicator research includes the
development of methods to diagnose the presence of multiple chemical stressors in mixtures using
advanced molecular methods - DNA microarrays.
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Gregory P. Toth
Dr. Toth is the Branch Chief of the Molecular Ecology Research Branch at the U.S. EPA National
Exposure Research Laboratory (NERL) Ecological Exposure Research Division (EBRD) in
Cincinnati, OH. He has a Ph.D. in Biological Chemistry from the University of Cincinnati College
of Medicine, Department of Biological Chemistry (1982) and a BS in Chemistry from John Carroll
University (1975). Dr. Toth was an Albert J. Ryan Fellow (1976-1981) and received a National
Research Council Research Associateship (1985-1986). Dr. Toth has been published in numerous
journals and publications including: Molecular Ecology, Electrophoresis, the American Journal of
Botany, and Environmental Toxicology and Chemistry.
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DEVELOPMENT OF CHEMICAL METHODS TO CHARACTERIZE EXPOSURE TO
EDCS IN THE NEUSE RIVER BASIN
Myriam Medina-Vera
U.S. Environmental Protection Agency
NERL HEASD EMMB
79 T.W. Alexander Drive (MD-44)
Research Triangle Park
Durham, NC 27711
(919) 541-5016
(919) 541-3527 fax
Medina-Vera.Myriam(Sjepa.gov
To develop a quantitative health and environmental risk assessment of endocrine disrupting
compounds (EDCs), information on exposures is essential. A full exposure assessment has complex
requirements that require preliminary information to direct further research in this area. Such
research begins with refining the biological and chemical methods for selected endocrine disrupting
compounds in fresh and brackish, polluted and unpolluted waters, sediment, soil and selected aquatic
organisms. Accurate characterization of the exposures can be done by using valid methods that are
sensitive and reliable. Adequate methods provide tools that will help with the understanding of
pathways of exposure, fate and transport of selected endocrine disrupters. Identification of data gaps
stressed the need for better methods in the identification and quantitation of EDCs such as
alkylphenols and selected toxic metals.
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Myriam Medina-Vera
Dr. Medina-Vera has a BS in Chemistry and Mathematics (double major) and a Ph.D. in Analytical
Chemistry with minors in Physical Chemistry and Inorganic Chemistry from the University of Puerto
Rico. She has been working for EPA since 1991. Her research areas include Photochemistry of
PAHs on participate matter, Pyrolysis of PAHs, and analysis alkylphenols. She is currently the chief
of NERL's Exposure Methods and Monitoring Branch in RTP.
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Monitoring Endocrine Disrupting Compounds in Aquatic Ecosystems in the United States
Steven L. Goodbred,
U.S. Geological Survey, Placer Hall, 6000 J Street
California State University
Sacramento, CA 95821
(916) 278-3097
goodbred@usgs.gov
Several Federal Agencies have monitoring programs that include suspected endocrine disrupting
compounds (EDCs); however no systematic monitoring of only EDCs is routinely done in the United
States. The U.S. Geological Survey's National Water Quality Assessment Program (NAWQA) was
established to assess status and trends of surface-water and ground-water throughout streams, rivers,
and aquifers of the US (Gilliom et al. 1995). Fourty-four EDCs have been monitored in water, bed
sediment, and fish since 1991 at 1363 sites. Results have shown several suspected EDCs like atrazine
are widespread in US streams and their occurrence appears related to land and chemical use. Some
EDCs like PCBs are more frequently detected and at higher concentrations in urban streams while
others like p,p' DDE in fish are found in much higher concentration in agricultural land use. At
many sites the detection of breakdown products and metabolites was 10 to 25 times the concentration
of the parent compound.
Although pesticides are at low levels (
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Goodbred, S., R.Gilliom, T.Gross, N. Denslow, W.Bryant, and T.Schoeb, 1997, Reconnaissance of
175-estradiol, 11-ketotestosterone, vitellogenin, and gonad histopathology in common carp of United
States streams-Potential for contaminant-induced endocrine disruption: U.S. Geological Survey Open
File Report 96-627, 47p.
THE SPEAKER'S BIO IS UNAVAILABLE. PLEASE SEE THE SPEAKER WITH
COMMENTS AND/OR QUESTIONS
24
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REGION 5: ONGOING ENDOCRINE DISRUPTOR EFFORTS
Peter Howe, Al Alwan, John Dorkin, George Azevedo, Mari Nord, Marc Tuchman,
Dennis Wesolowski, Babu Paruchuri, Lawrence Zintek*
U.S. EPA Region 5
77 West Jackson Blvd.
Chicago, IL 60604
(312) 353-9067
(312) 886-2591 Fax
zintek.lawrence@epa.gov
Region 5 efforts are focused on the alkylphenols such as; nonylphenol, nonylphenol ethoxylates,
nonylphenol carboxylates and octylphenol. Our interest is to discover if these chemicals are present
in our region and if so are they present at a concentration harmful to the environment. The region was
determined to have the need for standards of known purity fulfilled by getting them synthesized and
made commercially available. The material presented includes several studies demonstrating that
effluent concentrations of chemicals of interest are above the effect level in water bodies receiving
effluent from Publicly Owned Treatment Works (POTW=s). Water, sediment and fish samples were
taken and analyzed from different river systems in Region 5. Some data were collected through
funding of studies by USGS and USDA.
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Lawrence Zintek
Dr. Lawrence Zintek is with the U.S. EPA Region 5, Central Regional Laboratory. Prior to working
at the U.S. EPA, he was an instructor at Benedictine University from 1995-2000. Dr. Zintek has a
Ph.D. in Chemistry from the University of Iowa and a BS in Biochemistry from Illinois Benedictine
College.
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RESIDENTIAL INDOOR AIR AND DUST MEASUREMENTS FOR PHTHALATES AND
OTHER ENDOCRINE DISRUPTING COMPOUNDS
Ruthann Rudel1*, Julia G. Brody1, David Camann2, Alice Yau2, John D. Spengler3
Silent Spring Institute, 29 Crafts Street, Newton, MA 02458
(617) 332-4288 x!4, (617) 332-4284 fax
Rudel@SilentSpring.org
2Southwest Research Institute, 6220 Culebra Road,PO Drawer 28510
San Antonio, TX 78228
(210) 522-5947, (210) 522-5938 fax
3Harvard School of Public Health, 665 Huntington Ave,Boston, MA 02115
(617) 432-1255, (617) 432-4122 fax
jspengle@hsph.harvard.edu
In order to characterize exposures to chemicals of interest for research on breast cancer and other
hormonally mediated health outcomes, residential air and dust samples were analyzed for up to 93
target compounds that 1) have been identified as animal mammary carcinogens or hormonally active
chemicals and 2) are used in commercial or consumer products or building materials. Selected
phthalates, pesticides, parabens, PAHs and PCBs were extracted and analyzed by GC/MS-SIM.
Phenolic compounds including nonylphenol, octylphenol, bisphenol A, and the methoxychlor
metabolite HPTE were extracted, derivatized, and analyzed by GC/MS-SIM (Rudel et al., 2001). In
data from the first 30 of 120 homes sampled on Cape Cod, MA, 44 of 68 target compounds were
detected in at least one air sample and 68 of 93 were detected in at least one dust sample.
In these 30 air samples, diethyl phthalate was detected at the highest concentrations (range 128-2,113
ng/m3), followed by dibutyl phthalate (56-851 ng/m3), di (2-ethylhexyl) phthalate (DEHP) (57-562
ng/m3) and diisobutyl phthalate (18-986 ng/m3). Six other phthalates were also detected. It is of
interest that the most abundant phthalates in indoor air are also most abundant in human urine
samples collected by CDC for a reference population of US adults (CDC 2001). Other EDCs present
in air at highest concentrations included nonylphenol (48-416 ng/m3) and the disinfectant o-phenyl
phenol (43-958 ng/m3). Pesticides detected in over half the air samples tested included propoxur,
chlordane, pentachlorophenol, heptachlor, diazinon, and chlorpyrifos.
In 30 dust samples, the most widely used phthalates were most abundant (NTP-CEHR 2000). Thus
DEHP was present at highest concentrations (range 125-1082 ?g/g), followed by BBP (9-749 ?g/g)
and diisononyl phthalate (
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Rudel, R.A.; Brody, J.G.; Spengler, J.D.; Vallarino, J.; Geno, P.W.; Sun, G.; Yau, A.; Journal of the
Air and Waste Management Association 2001, 51, 499-513.
National Toxicology Program Center for the Evaluation of Risks to Human Reproduction. NTP-
CERHR Expert Panel reports on seven phthalate esters, http://cerhr.niehs.nih.gov, 2000.
Centers for Disease Control and Prevention-National Center for Environmental Health. National
Report on Human Exposure to Environmental Chemicals, www. cdc. gov/nceh/dls/report. 2001.
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Ruthann Rudel, M.S.
Ruthann Rudel is a senior environmental toxicologisi at Silent Spring Institute, with experience in
toxicology, risk assessment and exposure assessment. One of Ms. Rudel's areas of research is the
potential health effects of exposure to compounds that affect the endocrine system. She oversees the
environmental assessment portion of the Cape Cod Breast Cancer and Environment study, a multi-
disciplinary research effort funded by the Massachusetts Department of Public Health to study the
elevated breast cancer incidence on Cape Cod. Ms. Rudel is the author of many articles in scientific
journals, including a recent paper in Journal of Air and Waste Management Association about the
identification of hormonally active agents and animal mammary carcinogens analyzed in commercial
and residential air and dust samples. Ms. Rudel has also published articles in Environmental Science
and Technology describing methods and results for testing groundwater for estrogenic phenols and a
paper in Environmental Health Perspectives identifying issues in risk assessment for estrogenic
chemicals. She has published journal articles and book chapters on regulatory toxicology, metals
risk assessment, indoor-air risk assessment, and other subjects. She serves on the Science Advisory
Board of the Massachusetts Toxics Use Reduction Institute. She has a B.A. in chemistry and
neuroscience from Oberlin College, and an M.S. in environmental management and policy from
Tufts University.
Silent Spring Institute, in Newton, MA, is a nonprofit scientific research organization dedicated to
identifying the links between the environment and diseases that affect women, especially breast
cancer.
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s Tuesday, January 29, 2002s
Risk Management Approaches
Moderator: Andy Avel, U.S. EPA, National Risk Management Research Laboratory
4:00 PM EPA's Risk Management Evaluation ofEDCs
Gregory Sayles, U.S. EPA, National Risk Management Research Laboratory
4:40 PM Using Bioassays to Evaluate the Performance of Risk Management
Techniques
Carolyn Acheson, U.S. EPA, National Risk Management Research
Laboratory
5:10PM Adjourn for the day
31
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32
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EPA's RISK MANAGEMENT EVALUATION OF EDCs
Gregory D. Sayles
U.S. Environmental Protection Agency, Office of Research and Development
National Risk Management Research Laboratory, Cincinnati, OH 45268
(513) 569-7607, (513) 569-7105 fax
sayles.gregory@epa.gov
EPA's National Risk Management Research Laboratory (NRMRL) is developing a methodology to
assess the current state of risk management of environmental problems. The findings from carrying
out this methodology on a particular environmental challenge have been termed the Risk
Management Evaluation (RME). In the last 2 to 3 years, pilot / demonstration RMEs were
constructed for several environmental issues including endocrine disrupting chemicals (EDCs).
Based on the experience gained in producing these pilot RMEs, a protocol to guide the writing of
future RMEs is currently being finalized in NRMRL. The purpose of this presentation is to
summarize the content of the RME for EDCs, which is in final draft form at this time.
The RME for EDCs presents in a succinct manner the current understanding of risk management of a
short list of known or likely EDCs. The RME serves several purposes. First, the document identifies
currently available risk management approaches that were developed for other purposes, but appear
useful in managing the risk of EDCs. Second, the document indicates where new risk management
approaches are needed. Thus, the RME will be useful (1) to inform risk managers such as regulators
on what technical approaches are currently available for managing EDCs risk, (2) to educate the
public about what risk management approaches are available now, (3) to motivate environmental
consultants/engineers to review current skills or to develop new skills applicable to managing
exposure to EDCs, and (4) to guide risk management researchers, such as NRMRL, in planning EDC
risk management research programs.
Since health effects, exposure, risk assessment and risk management knowledge change with time,
the RME must be updated regularly. Thus, we consider the RME as a living document to be updated
as needed, approximately yearly, and downloadable from an EPA web site.
The presentation will discuss the specific content of version 1.0 of the RME for EDCs. This version
will include discussion of the following known or likely EDCs: alkylphenol ethoxylates and related
chemicals, natural, veterinary, and pharmaceutical steroid hormones, DDT and DDE, PCBs, and
chlorinated dioxins and furans.
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Gregory Sayles
Gregory Sayles is a chemical engineer with EPA=s National Risk Management Research Laboratory
in Cincinnati, OH and leads the endocrine disrupting chemicals risk management research program.
Dr. Sayles earned B.S., M.S., and Ph.D. degrees in chemical engineering from the California Institute
of Technology, the University of California at Davis, and North Carolina State University,
respectively. Dr. Sayles has worked for NRMRL for eleven years, conducting research mostly on
bioremediation of contaminated soils and sediments.
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USING BIOASSAYS TO EVALUATE THE PERFORMANCE OF RISK MANAGEMENT
TECHNIQUES
Carolyn Acheson* and Gregory Sayles
U.S. Environmental Protection Agency, Land Remediation and Pollution Control Division,
National Risk Management Research Laboratory
26 W. Martin Luther King Dr
Cincinnati, OH 45268
(513) 569-7190, (513) 569-7105 (fax)
acheson.carolyn@ epa.gov
Often, the performance of risk management techniques is evaluated by measuring the concentrations
of the chemicals of concern before and after risk management efforts. However, using bioassays and
chemical data provides a more robust understanding of the effectiveness of risk management
strategies. For example, bioassay testing evaluates the aggregate effect of the environmental sample
on the reporting organisms, and thus, includes aspects such as environmental matrix effects,
sorption/desorption behavior, bioavailability, and chemical mixture interactions. As a result,
bioassay testing can demonstrate changes in toxicity rather than inferring risk reduction from
chemical concentrations. When bioassays are used to evaluate samples following risk management
techniques, increased responses are observed in some cases. These increased responses may be due
to incomplete treatment or toxicity introduced through process amendments. When these types of
problems are identified through bioassay testing, risk management techniques can be altered to
correct the problem. The combination of chemical and bioassay data has been helpful in evaluating
risk reduction technologies treating soils contaminated with hazardous wastes. Due to the limited
knowledge about the endocrine activity of various chemicals and their degradation products,
bioassays are even more important in evaluating the performance of EDC risk management
treatments.
Several EDC bioassays have been developed to identify EDCs or to characterize EDC health effects
and exposure levels. To find an assay suitable for risk management projects, assays from the peer
reviewed literature were evaluated based on: reported sensitivity, range of applications and
chemicals studied, acceptance in the academic community, the details of the assay protocol, and the
EDSTAC Tier I Screening Battery recommendations. Practical concerns such as cost, time,
equipment and space needs, and personnel skills were also included in the evaluation process. Since
most current risk management projects are concerned with estrogenic compounds, androgenic and
thyroid assays were not considered at this time. Based on these criteria, the Yeast Estrogen
Screening assay was selected as the first assay to be adapted for risk management projects in
NRMRL.
In addition to discussing the benefits of including bioassay testing in evaluation of risk management
effectiveness and the assay selection process, this presentation will discuss adapting a bioassay for
NRMRL projects and a few hypothetical case studies illustrating the use of EDC bioassays to
evaluate the effectiveness of risk management techniques.
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Carolyn Acheson
Carolyn Acheson has been trained as a chemical engineer with emphasis in microbiology. She was
awarded a Bachelor's of Chemical Engineering from the University of Delaware and a Ph.D. from
Cornell University. She has worked at the U.S. EPA's National Risk Management Research
Laboratory (NRMRL) since 1994. While atNRMRL, Dr. Acheson has studied the bioremediation of
hazardous wastes particularly soils contaminated with polycyclic aromatic hydrocarbons. In many of
these studies, remedial technology performance was evaluated using bioassays as well as the more
traditional, chemical data. Based on her experience with soil toxicity, Dr. Acheson has recently
begun a project to adapt EDC bioassays for use in evaluating remedial treatments for EDC
contaminated matrices.
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»Wednesday, January 30, 2002s
Drinking Water Treatment
Moderator: James Goodrich, U.S. EPA, National Risk Management Research
Laboratory
8:35 AM Use of Granular Activated Carbon and Powdered Activated Carbon for the
Removal ofEDCs from Drinking Water: A User's Guide
John Cicmanec, U.S. EPA, National Risk Management Research Laboratory
9:05 AM Evaluation of Drinking Water Treatment Technologies for Removal of
Endocrine Distrupting Compounds
Kathleen Schenck, U.S. EPA, National Risk Management Research
Laboratory
9:20 AM Risk Management of Endocrine Disrupting Chemicals (EDCs) in Drinking
Water
Frederick Pontius, Pontius Water Consulting
9:50 AM Break
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38
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USE OF GRANULAR ACTIVATED CARBON AND POWDERED ACTIVATED CARBON FOR
THE REMOVAL OF EDC'S FROM DRINKING WATER: A USER'S GUIDE
John L. Cicmanec, DVM, MS
U.S. EPA ORD NRMRL STD SAB
26 W. Martin Luther King, Jr. Drive
Cincinnati, OH 45268
(513) 569-7481, (513) 569-7585 fax
cicmanec.john@epa.gov
Recently, public concern has increased regarding industrial and environmental substances that may
have adverse hormonal effects in human and wildlife populations. Although the list of potentially
harmful substances is still being compiled and more sophisticated laboratory tests for detection of
endocrine disrupting chemicals (EDCs) are being developed, an initial list of known EDCs has been
made and an array of drinking water treatment processes has been evaluated for their ability to
remove EDCs. Alkylphenols, bisphenol A, phthalates, poly chlorinated biphenyls, dioxins,
dibenzofurans as well as the pesticides methoxychlor, endosulfan, and DDT have been included in
the initial list. In addition to the conventional water treatment processes of sedimentation,
coagulation, and filtration, we have also considered the potential effects of granular activated carbon,
powdered activated carbon, nanofiltration, reverse osmosis and air stripping for the removal of
EDCs. Our findings indicate that granular activated carbon (GAC) and powdered activated carbon
(PAC) are the most effective processes for the removal of the selected group of EDCs. The
laboratory data that was used in the Freundlich equation for determining the efficiency of GAC for
EDC removal will be presented. Additional analyses of the effectiveness for removing
ethinylestradiol and melegesterol acetate through the use of GAC and PAC will also be presented.
39
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John Cicmanec
Dr. Cicmanec is a graduate of the Kansas State University veterinary college and he has completed
post-doctoral training at the University of Michigan Medical School. His clinical experience
includes two years in large animal dairy practice and 15 years experience in biomedical research with
non-human primates. This work involved infectious disease studies and part of this work included
international scientific exchange programs with the Soviet Union, Bolivia and Peru. He has been at
EPA for 16 years and his primary responsibilities have involved management of animal research
colonies and noncancer risk assessment. He has been responsible for the risk assessments of PCBs
and methylmercury and has participated in the review of 200 other chemical risk assessments. He
was on the scientific team that wrote the Agency's Strategic Plan for Children's Health and his
present research interests include endocrine disrupting chemicals and animal pathogens that are
transmitted to humans.
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EVALUATION OF DRINKING WATER TREATMENT TECHNOLOGIES
FOR REMOVAL OF ENDOCRINE DISRUPTING COMPOUNDS
Kathleen Schenck l", Thomas Speth l, Radha Krishnan2, Barry Pepich2, Steve Wendelken2,
Laura Rosenblum2, Kent Mitchell3, and David Warshawsky3
^.S. EPA, National Risk Management Research Laboratory
26 West Martin Luther King Drive, Cincinnati, OH 45268
513-569-7947, fax: 513-569-7185, schenck.kathleen@epa.gov
2 IT Corporation
11499 Chester Road, Cincinnati, OH 45246
513-569-7063, fax: 513-569-7707, krishnan.radha@epa.gov
3 University of Cincinnati, College of Medicine, Department of Environmental Health
P.O. Box 670056, Cincinnati, OH 45267
513-558-5428, fax: 513-558-5561, mitchekm@email.uc.edu
Many of the chemicals identified as potential endocrine disrupting compounds (EDCs) may be
present in surface or ground waters used as drinking water sources due to their introduction from
domestic and industrial sewage treatment systems and wet-weather runoff. In order to decrease
the risk of potential adverse health effects associated with the presence of EDCs in drinking
water, two basic strategies exist. One is to protect source waters from contamination by EDCs.
The other is to remove EDCs, which may be present in source waters, during the drinking water
treatment process. This project addresses the latter approach by evaluating the removal of
several EDCs by various drinking water treatment processes. The compounds to be evaluated
are all steroid hormones: estradiol; estriol; ethynylestradiol; progesterone; testosterone and
dihydrotestosterone. In the future, a group of nonylphenolic compounds will also be added.
This project is divided into four parts. The first is the development of an analytical method to
identify and quantify the analytes. The approach will include a solid phase extraction step
followed by liquid chromatography/mass spectroscopy (LC/MS). The second is the application
of a reporter gene assay, the MVLN assay, to evaluate the removal of estrogenic activity from
the water samples. This assay uses a human breast cell line which has been transfected with the
firefly luciferase reporter gene. This assay should detect the presence of compounds that have
estrogenic activity, including those that may be missed analytically due to structural changes in
the target compounds during treatment. Once the analytical and MVLN assays are in place,
bench-scale evaluations of various drinking water treatment processes will be conducted. These
will include conventional treatment, granular activated carbon, softening and nanofiltration. For
each of these processes, pilot-scale evaluations may be conducted, if warranted.
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Kathleen Schenck
Kathy Schenck is an environmental scientist with the Environmental Protection Agency, National
Risk Management Research Laboratory. She has worked in the drinking water area for more than 20
years. She has a Masters Degree in biology from the University of Cincinnati.
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RISK MANAGEMENT OF ENDOCRINE DISRUPTING CHEMICALS (EDCs)
IN DRINKING WATER
Frederick W. Pontius, P.E.
President
Pontius Water Consultants, Inc.
P.O. Box 150361
Lakewood, CO 80215-0361
(303) 986-9923, (303)716-7310 fax
fredp@pontiuswater.com
INTRODUCTION
Endocrine disrupting chemicals (EDCs) have emerged within the last decade as an important
environmental issue. The implications for drinking water and water suppliers have only been
seriously considered within the last several years. The U.S. Environmental Protection Agency (US
EPA) Office of Ground Water and Drinking Water (OGWDW) is including EDCs and endocrine
effects as it considers setting future drinking water regulations.
An April 2000 AWWA Research Foundation/Water Environment Research Foundation/ WaterRe-
use Foundation workshop on Endocrine Disrupters and Pharmaceuticals in Drinking Water (Weyer
and Riley, 2001) provided an opportunity for water suppliers to become informed about the science,
potential health risks, regulation, and control of EDCs in drinking water. Water suppliers are
considering how to minimize their customers risks to EDCs in drinking water.
DRINKING WATER REGULATION
Historically, U.S. EPA has considered endocrine effects in the regulation of drinking water
contaminants when setting Maximum Contaminant Level Goals (MCLGs). Many contaminants that
are known or suspected to be EDCs are already regulated in drinking water. Should new information
regarding their endocrine effects become known, the Safe Drinking Water Act (SOWA) requires
review and revision of existing regulations as needed at least every six years.
The endocrine effects of unregulated drinking water contaminants will be considered as part of the
evaluation of contaminants listed on the Candidate Contaminant List (CCL). U.S. EPA will be
making determinations for at least 5 contaminants on the CCL regarding whether regulation is
needed, every 5 years, starting in 2001. Endocrine effects will be considered in these determinations.
A STRATEGY FOR WATER SUPPLIERS
Regardless of the regulatory actions taken by U.S. EPA, water suppliers must respond to customer
concerns. As awareness of the potential effects of EDCs increases, customers naturally wonder
whether EDCs are present in their drinking water. Many utilities have been actively addressing how
they can minimize risk to customers from EDCs and communicate risks effectively with their
customers. A simple risk management process can provide a framework for water suppliers to begin
to address the complex issues associated with EDCs in drinking water.
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REFERENCE
Weyer, P., and Riley, D. Endocrine Disrupters And Pharmaceuticals in Drinking Water. AWWA
Research Foundation, Denver, Colo. (2001).
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Frederick W. (Fred) Pontius
Fred Pontius is President of Pontius Water Consultants, Inc., in Lakewood, Colorado, a small
company providing drinking water related professional engineering services to water utilities,
consultants, industry, businesses, and regulatory agencies. He has 23+ years of experience in public
water supply, including water quality control, design, research, and government affairs. He has
prepared compliance and water quality control plans for large and small water systems, with
consideration of endocrine disrupting chemicals (EDCs) and Pharmaceuticals. Fred has written
several books and over 150 technical articles on drinking water regulation, compliance, and water
quality control, including EDCs. He is a frequent conference speaker, and has conducted training
seminars for USEPA, the American Water Works Association, the National Park Service, and the
Government Institutes. He is a registered professional engineer in Colorado and Ohio. In his spare
time, Fred is a doctoral candidate in environmental engineering at the University of Colorado—
Boulder, conducting research on membrane treatment.
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»Wednesday, January 30, 2002s
Concentrated Animal Feed Operations
Moderator: Laurel Staley, U.S. EPA, National Risk Management Research Laboratory
10:10 AM Potential of Confined Animal Feed Operations (CAFOs) to Contribute
Estrogens to the Environment
Steven Hutchins, U.S. EPA, National Risk Management Research Laboratory
10:40 AM Investigations of Sorption and Transport of Hormones and Animal
Pharmaceuticals: Initial Laboratory Results
Suresh Rao, Purdue University
Linda Lee, Purdue University
11:00 AM Fate of the Endogenous Hormones 17K-Estradiol and Testosterone in
Composted Poultry Manure and their Sorption and Mobility in Loam Soil and
Sand
Heldur Hakk, USDA, Agricultural Research Service
11:30 AM Lunch
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POTENTIAL OF CONFINED ANIMAL FEED OPERATIONS (CAFOS) TO CONTRIBUTE
ESTROGENS TO THE ENVIRONMENT
Stephen R. Hutchinsl", James N. Dumont2, David M. Janz2, G. Peter Breidenbach3,
Dennis D. Fine3
^.S. EPA NRMRL, Subsurface Protection and Remediation Division
Robert S. Kerr Environmental Research Center, P.O. Box 1198, Ada, OK 74820
(580) 436-8563, (580) 436-8703 fax
hutchins.steve@epa.gov
Department of Zoology, Oklahoma State University
430 Life Sciences West, Oklahoma State University, Stillwater, OK 74078
(405) 744-9683, (405) 744-7824fax
dumontj @okstate.edu
(405) 744-7593, (405)744-7824 fax
djanz@okstate.edu
3ManTech Environmental Services, Inc.
Robert S. Kerr Environmental Research Center. P.O. Box 1198, Ada, OK 74820
(580) 436-8668, (580) 436-8501 fax
breidenbach.peter@epa.gov
(580) 436-8669, (580) 436-8501 fax
fine.dennis@epa.gov
Abstract
Confined Animal Feed Operations (CAFOs) are a growing industry, with a trend towards fewer
operations with higher concentrations of animals. Animals are fed and/or treated with many different
types of Pharmaceuticals, including antibiotics and hormones, which can end up in waste products.
Some of these chemicals, in particular the natural and synthetic hormones, can exert endocrine-
disrupting activity in the environment. The overall objective of this research is to evaluate whether
animal wastes contain chemicals with EDC activity, and whether these chemicals are sufficiently
persistent so as to pose a risk to receiving ground and surface waters.
Approach
There are many unknowns regarding the potential for CAFOs to contribute EDCs to the environment,
due to the variety of CAFO operations, the diverse natures of potential EDCs themselves, and the
different types of environmental receptors that could be affected. Hence, several projects to address
these different issues are anticipated. Two projects have thus far been initiated, one to evaluate EDC
activity from different types of CAFOs, and the other to measure levels of estrogens in swine waste
effluents. These are described below.
EDC Activity in Different Types of CAFO Lagoons
This research is being conducted by Oklahoma State University under a cooperative agreement, and
is designed to evaluate swine, dairy, and beef CAFO lagoons for EDC activity. Three analyses will
be used to assess for the presence of EDCs. The first, the Xenopus Tail Resorption Assay (XTRA),
measures the tail resorption rate prior to metamorphosis as a function of the presence of thyroid
disrupters. The second measures plasma vitellogenin concentrations in male frogs using an enzyme-
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linked immunosorbent assay (ELISA) to detect the presence of estrogenic compounds. The third
measures plasma testosterone and 17B-estradiol concentrations as an indicator of alterations in
reproductive endocrine homeostasis.
This research is ongoing. Preliminary results show that, although the swine effluent lagoon is quite
toxic, none of the lagoons have exhibited significant EDC activity, at least based on these bioassays.
It's quite possible that EDC activity is truly insignificant in CAFO lagoons. However, it may also be
that these lagoons are insufficiently representative of large-scale commercial operations, or that there
may be EDC effects under long-term exposure that are not detected with these bioassays.
Analysis of Estrogens in Swine Waste Effluents
Analysis of environmental samples for low levels of hormones is a rapidly-expanding field, and there
have been many new developments in adapting current analytical techniques for these compounds.
This work is being conducted by ManTech Environmental Services under a contract, and the primary
objective is to optimize both ELISA and LC/MS/MS analytical procedures for analysis of complex
wastewaters for estrogenic compounds, including 17B-estradiol, estrone, estriol, and ethinyl estradiol.
The goal is to use ELISA for screening of environmental samples for estrogenic activity, and then to
confirm the presence of the individual estrogens in samples that test positive from ELISA. This work
is being coordinated through SPRD so that different swine CAFO waste samples can be obtained,
screened by ELISA, and then analyzed by LC/MS/MS to determine the concentrations of
environmental estrogens.
This research is also ongoing. Preliminary results show that the analyses work well for ground water
samples; detection limits are on the order of 0.05 ng/L for ELISA screening based on 17B-estradiol,
and 2 ng/L for estrogen separation and identification by LC/MS/MS. However, ELISA screening of
swine lagoon effluents often yield 17B-estradiol concentration estimates orders of magnitude higher
than what can be confirmed by LC/MS/MS. HPLC clean-up of extracts reduces this interference, but
does not eliminate it. LC/MS/MS analysis of several swine lagoon effluents shows that estrone is the
most predominant estrogen.
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Stephen R. Hutchins
Stephen R. Hutchins is a Research Environmental Scientist at EPA's National Risk Management
Research Laboratory, Subsurface Protection and Remediation Division, located at the Robert S. Kerr
Environmental Research Center in Ada, Oklahoma. Dr. Hutchins has a B.S. in Biology from the New
Mexico Institute of Mining and Technology and a Ph.D. in Environmental Science from Rice
University. His research interests are directed towards bioremediation of contaminated aquifers by
indigenous subsurface bacteria, and his past work focused on laboratory and field evaluation of
biorestoration of fuel-contaminated aquifers under anaerobic conditions. His current research project,
funded under the NRMRL Laboratory Director's Special Grants Program, has been to evaluate the
potential for ground water contamination from swine CAFOs.
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INVESTIGATIONS OF SORPTION AND TRANSPORT OF HORMONES AND ANIMAL
PHARMACEUTICALS: INITIAL LABORATORY RESULTS
Linda S. Lee1, P. Suresh C. Rao2
1 Agronomy Department, Purdue University
West Lafayette, IN 47907
(765) 494-8612
(765) 496-1107 fax
lslee(S),purdue.edu
2School of Civil Engineering & Agronomy Department, Purdue University
West Lafayette, IN 47907
765-496-6554
pscr@purdue.edu
With joint funding from U.S. EPA and the College of Agriculture, Purdue University, a series of
laboratory studies has been initiated to evaluate sorption, transport and transformations of several
compounds that have the potential to function as endocrine disrupters. The groups of test compounds
investigated to date include: two hormones (estrodiol and testosterone), and three animal antibiotics
(carbadox, tylosin, bacitracin) that are also used at sub-therapeutic levels as growth promoters in
swine production. Initial project activities focused an extensive literature search to identify available
environmental fate data and protocols for LC or GC analysis. Initial laboratory studies are focused on
determining equilibrium adsorption isotherms using several Midwestern US soils and selected
samples from a collection of soils and freshwater sediments that were used in past EPA
investigations. These soil and sediments represent a wide range in sorbent characteristics, including
pH, organic carbon content, clay content//type, CEC, geographic locations, etc. Batch isotherm data
is being used to interpret the likely sorption mechanisms, and potential for leaching and retardation.
Companion laboratory experiments are being conducted to evaluate transport and transformations
under steady, saturated water flow conditions in columns packed with soils or sediment samples. The
measured breakthrough curves are being interpreted using temporal moment analysis (to estimate
mass conservation and retardation), and by fitting to existing transport models (to estimate
retardation and non-equilibrium sorption parameters). Designs for a future field-scale test to examine
retardation and degradation of the test compounds are also being developed based on preliminary
tracer studies currently underway.
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Linda S. Lee
Linda S. Lee is Professor of Environmental Chemistry & Soil Science in the Agronomy department
at Purdue University. Since receiving a PhD from the University of Florida in 1991, she has been on
the faculty at Purdue University. Her current research interests include remediation of soils
contaminated with organics and metals, and basic research in environmental chemistry. She teaches
courses in environmental organic chemistry and remediation science.
P. Suresh C. Rao
P. Suresh C. Rao is the Rieth Distinguished Professor of Environmental Engineering in the School of
Civil Engineering, with a joint appointment in the Agronomy department, at Purdue University. Prior
to arriving at Purdue two years ago, he was on the faculty for 25 years at the University of Florida.
His current research interests include remediation science and engineering as well as watersheds and
water quality. He teaches courses in subsurface hydrology and remediation.
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FATE OF THE ENDOGENOUS HORMONES 17B-ESTRADIOL AND TESTOSTERONE IN
COMPOSTED POULTRY MANURE AND THEIR SORPTION AND MOBILITY IN LOAM SOIL
AND SAND
Heldur Hakk1*, Patricia Millner 2, and Gerald Larsen1
Colleen Pfaffl, Barb Magelky1, and Frank Casey3
1 USDA-ARS Biosciences Research Lab, Fargo, ND
2 USDA-ARS Soil Microbial Systems Lab, Beltsville, MD
3 North Dakota State University, Dept. Soil Science, Fargo, ND
701-239-1293; 701-239-1430 fax
hakkh@fargo.ars.usda.gov
Relatively little attention has been paid to the fate and environmental impact of the potent hormones,
17B-estradiol and testosterone, eliminated in animal and human waste. Laying chickens can excrete
these hormones at 500 and 250 ng/g dry manure/day, respectively l. Furthermore, estradiol has a
100-fold greater affinity for the estrogen receptor than do man-made estrogenic substances of current
environmental concern 2. Land application of manure has a considerable potential to affect the
environment adversely. Composting is known to effectively remediate soils contaminated with toxic
organic compounds 3. Thus, the present study was undertaken to quantitatively assess whether 17B-
estradiol and testosterone could be degraded during composting. Chicken layer manure was mixed
with plant material to achieve a C:N ratio of 30:1, and place in 4 windrows (23 x 2 x 1.75 m). All
windrows were turned weekly. Commercial enzyme immunoassay kits were used to quantitate the
levels of 17B-estradiol and testosterone in the aqueous extracts of samples. The results demonstrated
that aerobic microorganisms degraded both hormones during the composting process. Composting
resulted in a steady decline in extractable estradiol and testosterone levels. These data suggest that
composting may be an environmentally friendly technology suitable for reducing the concentrations
of these endogenous hormones at concentrated animal operation facilities.
Transport and fate experiments of [14C] testosterone and 17B-estradiol were conducted by
individually applying each to columns packed with loam soil or sand. No radioactivity from either
hormone was found in the effluent of the loam soil column. Combustion analysis of the soil sections
showed that 80% and 96% of testosterone and estradiol, respectively, remained in the top five
centimeters of soil. A nonequilibrium sorption convective-dispersive transport model was applied to
the concentration profiles of the soil columns provided estimates of Kd values. Most of the
testosterone and estradiol (87% and 85%, respectively) was eluted from the sand columns,
respectively. Batch studies using the Glyndon silt-loam were conducted at 48, 96, and 178 h
incubation times and Freudlich isotherms were fit to the observed data. For both hormones, the
sorption isotherms were similar for all three incubation times and indicated that equilibration was
reached by 48 h. The Kd values indicated that these chemicals readily sorbed to the soil.
1 Shore, L.S., Gurevich, M., Shemesh, M., 1993, Bull. Environ. Contam ToxicoL, 51, 361-366.
2
Jobling, S. and Sumpter, IP., 1993, Aquatic Toxicology, 27, 261-272.
* US-EPA, 1998, Document EPA530-R-98-088, pp. 13-38.
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Heldur Hakk
Heldur Hakk has been with the USDA/ARS/Biosciences Research Laboratory for 23 years, and is a
Research Chemist in the Animal Metabolism-Agricultural Chemicals Research Unit. He obtained a
PhD in Biochemistry from North Dakota State University in 1997. His areas of research include:
mammalian metabolism of dioxins, PCBs, and brominated flame retardants, protein transport of
chemicals in mammals, effect of exposure to xenobiotics on circulating hormones, applications of
NMR to assess impact of foreign compounds to central metabolic pathways, and the fate of
hormones in the environment.
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»Wednesday, January 30, 2002s
Wastewater Treatment
Moderator: Marc Mills, U.S. EPA, National Risk Management Research Laboratory
12:45 PM Biological Fate of Estrogenic Compounds Associated with Sewage
Treatment A Review
Gregory Sayles, U.S. EPA, National Risk Management Research Laboratory
1:15 PM An Engineering Approach to Evaluate Estrogenic EDCs Fate During
Wastewater Treatment
Paul McCauley, U.S. EPA, National Risk Management Research Laboratory
1:30 PM Break
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BIOLOGICAL FATE OF ESTROGENIC COMPOUNDS ASSOCIATED WITH SEWAGE
TREATMENT
Gregory D. Sayles1*, Tamara Marsh2
^.S. Environmental Protection Agency, Office of Research and Development
National Risk Management Research Laboratory, Cincinnati, OH 45268
513-569-7607
513-569-7105 fax
sayles.gregory@epa.gov
2Elmhurst College, Dept. of Biology
190 Prospect Ave., Box 145
Elmhurst, IL 60126
(630) 617-3591
(630) 617-3735 fax
tamaram@elmhurst.edu
Considerable concern exists over the possibility that some man-made chemicals that mimic the
effects of hormones may have adverse effects on reproduction of wildlife and humans. Like natural
estrogens, these estrogenic compounds can bind to the estrogen receptor, thereby disrupting the
normal function of the endocrine system. Reproductive toxicity has been attributed to prolonged
exposure to environmentally relevant concentrations of estrogenic compounds. Two categories of
endocrine disrupting chemicals are thought to be the greatest contributors of environmental
contamination by estrogen mimicking chemicals. Naturally occurring hormones, such as 17?-
estradiol and estrone, as well as chemically synthesized forms, including the main component in oral
contraceptives, 17? -ethynylestradiol, by their very nature will make their way to sewage treatment
plants (STPs). In addition, alkylphenol polyethoxylates (APEs), used in the production of
surfactants, cleaning products, textiles, petroleum, pulp and paper, and pesticides can also mimic the
effects of hormones to the endocrine system. Like the hormone class of chemicals, these compounds
are present in sewage treatment plant influent. Endocrine disrupting chemicals, whether naturally
occurring or chemically synthesized, may be transformed or partially degraded during the processes
of aerobic and anaerobic sewage treatment. However, these chemicals or their derivatives are
accumulating in ecosystems impacted with sewage treatment effluent, indicating that they are not
being adequately degraded in the sewage treatment plant. Alkylphenol polyethoxylates have been
shown to lose portions of the polyethoxylate chain during sewage treatment, however, the most
prevalent resultant compound, nonylphenol (NP) is known to accumulate and cause toxicity in
aquatic organisms even at low concentrations. Thus, the degradation product poses more risk to the
environment than the parent compound. While less is known about the fate of the hormone
compounds, evidence suggests that these compounds are also accumulating in the environment and
causing reproductive failure in a variety of wildlife. This presentation will examine the current state
of knowledge concerning the microbial degradation of these categories of endocrine disrupting
chemicals, specifically their fate at the sewage treatment level as well as long-term fate once they are
released into the environment.
References
Hesselsoe, M., Jensen, D., Skals, K., Olesen, T., Moldrup, P., Roslev, P., Mortensen, O.K., and
Henriksen, K. 2001. Degradation of 4-Nonylphenol in Homogeneous and Nonhomogeneous
mixtures of soil and sewage sludge. Environ. Sci & TechnoL (In Press).
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Ternes, T.A., Kreckel, P., and Mueller, J. 1999. Behaviour and occurrence of estrogens in municipal
sewage treatment plants - II. Aerobic batch experiments with activated sludge. Sci. Total
Environ. 225: 91-99.
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Gregory Sayles
Gregory Sayles is a chemical engineer with EPA=s National Risk Management Research Laboratory
in Cincinnati, OH and leads the endocrine disrupting chemicals risk management research program.
Dr. Sayles earned B.S., M.S., and Ph.D. degrees in chemical engineering from the California Institute
of Technology, the University of California at Davis, and North Carolina State University,
respectively. Dr. Sayles has worked for NRMRL for eleven years, conducting research mostly on
bioremediation of contaminated soils and sediments.
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AN ENGINEERING APPROACH TO EVALUATE OF ESTROGENIC EDCS FATE
DURING WASTEWATER TREATMENT
Paul T. McCauley
U.S. EPA, National Risk Management Research Laboratory
26 West Martin Luther King Drive
Cincinnati, OH 45268
(513) 569-7444
(513) 569-7105 fax
McCauley.Paul@EPA.gov
Evidence is accumulating that environmental EDCs are affecting wildlife populations. Environmental
EDCs appear to have a range of effects. Some studies report unusual gene induction patterns in male
fish, while others have indicated that EDCs may result in hermaphrodism in native fish populations.
One source of environmental EDCs are POTWs. Desbrow et al. 1998, reported POTWs discharged
several estrogenic steroids including: estrone, 1 to 50 ng/L; 17 B-estradiol, 2 to 50 ng/L; and 17 a-
ethynylestradiol, 0 to 7 ng/L. At these levels, significant induction of vitellogenin was observed in
several fish species. Others have speculated that alkylphenols, three ethoxylates and metabolites may
have estrogenic effects.
We will be investigating the effectiveness of wastewater treatment plants and various steps in
wastewater treatment at removing estrogenic EDCs. Two conventional activated sludge pilot plants,
one employing anaerobic sludge digestion and the other aerobic sludge digestion are currently being
built. The pilot plants are to be identical in all respects except for the method of sludge digestion.
Because many POTWs today have stringent ammonia nitrogen discharge standards, the pilot plants
are designed to nitrify. In addition to facilitating EDC analytical methods development through the
provision of liquid stream samples of known EDC compound concentrations, these parallel flow
regimes will provide comparative information on the two most common in-plant means of stabilizing
and reducing the mass of waste sludges used in municipal wastewater treatment plants. The pilot
plant trains will consist of the following unit processes:
$ Primary clarifier (both trains)
$ Aeration tank (both trains)
$ Secondary clarifier (both trains)
$ Single-stage, high-rate anaerobic sludge digester (anaerobic digestion train only)
$ Aerobic sludge digester (aerobic digestion train only)
$ Sludge dewatering unit (both trains)
Chemical methods including solid phase extraction and GC/MS will be used to measure steroids and
smaller ethoxylate chain nonyl phenols and normal phase HPLC will be employed to analyze larger
ethoxylate chain nonyl phenols. Biological methods including mRNA (vitellogenin) induction
analysis in Fathead Minnow and a recombinant yeast assay will be used. These methods will be
employed to measure the removal rates of the various estrogenic EDC at several points in the pilot
plant wastewaster treatment process (including both liquid and solid waste streams). When steady
state results are achieved, the pilot plants will be adjusted to maximize estrogenic EDC removal.
After optimization is complete these analysis will be taken to the field at operating wastewater
treatment plants. We will evaluate these plants for Estrogenic EDC removal with attention to
optimization of the removal process. The final output of this research effort will be to make final
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recommendations for improved estrogenic EDC removal in publicly owned wastewater treatment
plant.
Desbrow et al. (1998) Environmental Science and Technology 32 (11): 1549-1556
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Paul T. McCauley
Dr. McCauley is a chemist with the U.S. EPA National Risk Management Laboratory (NRMRL_
Land Remediation and Pollution Control Division Treatment and Destruction Branch. He has a
Ph.D. from Ohio State University, College of Medicine Department of Pharmacology. He has
worked predominantly in bioslurry and bioventing vadose zone remediaiton. Dr. McCauley has
published papers in metals toxicity (pb, Ba, A, and orano-tins), kinetics, central nervous system,
endocrine, and general toxicology including: cardiovascular and hepatic toxicology.
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»Wednesday, January 30, 2002s
Other EDC Risk Management Challenges
Moderator: Andy Avel, U.S. EPA, National Risk Management Research Laboratory
1:45 PM Endocrine Disrupters from Combustion and Vehicular Emissions:
Identification and Source Nomination
Brian Gullett, U.S. EPA, National Risk Management Research Laboratory
2:00 PM Natural Recovery of PCB-Contaminated Sediments at the Sangamo-Weston /
Twelve Mile Creek/Lake Hartwell Superfund Site
Richard Brenner, U.S. EPA, National Risk Management Research Laboratory
James Lazorchak, U.S. EPA, National Exposure Research Laboratory
2:45 PM Program for the Identification and Replacement of Endocrine Disrupting
Chemicals
Douglas Young, U.S. EPA, National Risk Management Research Laboratory
3:00 PM Adjourn Workshop
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ENDOCRINE DISRUPTORS FROM COMBUSTION AND VEHICULAR EMISSIONS:
IDENTIFICATION AND SOURCE NOMINATION
Brian Gullett1*, Jeff Ryan1, Paul Lemieux1, Carolyn Acheson2, Michael DeVito3, James
Rabinowitz3, Sukh Sidhu4, Richard Striebich4, Joy Klosterman4
U.S. Environmental Protection Agency
JNational Risk Management Research Laboratory, Air Pollution Prevention and Control
Division (MD-65), Research Triangle Park, NC 27711
2National Risk Management Research Laboratory, Cincinnati, OH 45268
3National Health and Environmental Effects Research Laboratory, Toxicology Division,
Research Triangle Park, NC 27711
4University of Dayton Research Institute, 300 College Park, Dayton, OH 45469-0001
(919) 541-1534, (919) 541-0554 fax
gullett.brian@epa.gov
Exhaust samples from combustion and vehicular sources are being analyzed to provide initial
identification of endocrine disrupting chemicals (EDCs). The intent of this screening effort is to
provide discerning evidence for nominating sources for further EDC characterization. Conventional
sampling, advanced analytical methods, and bioassays are being used to provide initial
characterization of these samples for their compound identity and EDC activity. Our intent is to
sample and chemically characterize multiple combustion sources, consistent with the likelihood that
combustion source EDC exposure is not linked to any one source. For example, since body burdens
of polychlorinated dibenzodioxin/dibenzofuran do not appear to be elevated near traditionally
suspected sources (e.g., waste incinerators), it appears that exposure sources of dioxin are ubiquitous.
Sample fractionation will be coupled with chemical characterization, structure activity analyses, and
bioassay testing to nominate and identify potential EDC compounds in combustion emissions. Our
ability to provide this early source-specific EDC identification and characterization of combustion
sources will be a parallel activity with more extensive health risk analysis and exposure assessment.
In this manner, appropriate prioritization of EDC management options can be implemented prior to
definitive health effects conclusions.
A variety of combustion sources and fuel types will be opportunistically sampled, including diesel
fuels, fuel oil, biomass, natural gas, coal, refuse fuels, municipal and household wastes, and
hazardous waste. HPLC extract fractionation and estrogenicity testing using the Vitellogenin mRNA
Assay will be done on the samples with EPA/NEHERL and NRMRL/Ci. Multidimensional gas
chromatography (MDGC) by UDRI will characterize the sample unknowns and identify obscured
peaks.
Some samples have been analyzed via coventional GC- and MDGC-MS analysis and found to be
predictably complex. Currently we are investigating the analysis of individual compounds which
have structures similar to alkyl phenols (such as nonylphenol, a suspected endocrine disrupter) as
well as polycyclic aromatic hydrocarbons (metabolites of which can interact with the estrogen
receptor). Many compounds have been found that would have been difficult to detect in
conventional GC-MS because of coelution. For example, a methoxy alkyl phenol was found to
coelute with biphenyl under conventional conditions but was well separated using MDGC.
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Brian Gullett
Brian Gullett works in the area of trace combustion by-product formation, mainly chlorinated
dibenzodioxins and dibenzofurans (CDD/F), as well as fundamentals of mercury (Hg) sorption. His
current work includes development of the application of Jet Resonance-Enhanced MultiPhoton
lonization (REMPI) toward use as a trace toxics (including CDD/F) detector, determining the
reaction kinetics of CDD/F formation, and discernment of the kinetics and site-specific functional
groups of Hg reaction with sorbents. He has written over 125 papers; lectured in the US, Germany,
Australia, Sweden, and Korea; and served as conference/session chairs for international symposia on
CDD/F formation. He has twice been the recipient of the EPA's Federal Engineer of the Year award
and holds seven EPA Scientific and Technological Achievement Awards. Brian is Air Pollution
Editor for Environmental Engineering Science as well as a reviewer for a number of journals. In '95-
'96 his work included a one year assignment to the US Naval Surface Warfare Center where he
worked on shipboard incineration systems. He also serves as the Compliance pillar Co-Chair of the
Department of Defense's Strategic Environmental Research and Development Program (SERDP).
He has earned an A.B., an M.S. and Ph.D. in Environmental Engineering and, more recently, a
Master's of Engineering Management, all from Duke University. Needless to say, he is a basketball
fan.
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NATURAL RECOVERY OF PCB-CONTAMINATED SEDIMENTS AT THE
SANGAMO-WESTON/TWELVE MILE CREEK/LAKE HARTWELL SUPERFUND SITE
Richard C. Brenner1*, James M. Lazorchak2*, and Victor S. Magar3
1 U.S. EPA/NRMRL, 26 W. Martin Luther King Drive, Cincinnati, OH 45268
(513) 569-7657, (513) 569-7105 fax
b renner.richard @ep a.gov
2 U.S. EPA/NERL, 26 W. Martin Luther King Drive, Cincinnati, OH 45268
(513) 569-7076, (513) 569-7609 fax
lazorchak.jim@epa.gov
3 Battelle, 505 King Avenue, Columbus, OH 43201
(614) 424-4604, (614) 424-3667
magarv@battelle.org
Lake Hartwell is an artificial lake located in the northwest corner of South Carolina along the
Georgia state line. It was created between 1955 and 1963 when the US Army Corps of Engineers
constructed Hartwell Dam on the Upper Savannah River. It is the second largest lake in South
Carolina by volume and the third largest by surface area (56,000 acres).
The Sangamo-Weston plant was used for capacitor manufacturing from 1955 to 1978. The plant
used a variety of dielectric fluids in its manufacturing processes, including fluids containing PCBs.
Waste disposal practices at the site resulted in the cumulative discharge over the 23 years of plant
operation of an estimated 400,000 Ibs of PCBs into Town Creek, a tributary of Twelve Mile Creek,
which in turn is a tributary of Lake Hartwell. Source control implemented by EPA's Region 4 in the
1990s has successfully stopped the flow of PCBs into Lake Hartwell and its tributaries. With the
completion of source control, the PCB-contaminated Lake Hartwell ecosystem became a viable
candidate for natural recovery. Natural recovery was selected by the Region as the remedy of choice
for this site, as specified in its Record of Decision in 1994.
A field evaluation study was conducted in 2000 using a quantitative approach initiated by
EPA/NRMRL under contract to Battelle in cooperation with Region 4. The study characterized the
contaminated area and estimated the degree of recovery achieved in contaminated Lake Hartwell
sediments to date. This study has continued into 2001 with a second sediment sampling effort by
EPA/NRMRL and Battelle, combined with a fish toxicology evaluation by EPA/NERL. For both the
2000 and 2001 sampling efforts, sediment cores (T60 to »100 cm long x 5 cm diameter) were
collected from predetermined transect locations. The cores were divided into 5-cm segments. Each
segment was analyzed for PCBs to determine concentration profiles and 210Pb and 137Cs to age date
the sediment deposits. A total of 107 PCB congeners were quantified to characterize the source of
PCBs and the extent of dechlorination with sediment depth.
The data indicated that the net deposition of clean sediment has resulted in substantial burial of
contaminated sediment. In the six downgradient cores not affected by release of sediment from
upstream impoundments, the times to achieve three successively more stringent cleanup goals of 1.0,
0.4, and 0.05 mg/kg t-PCBs were estimated at 1 to 3, 2 to 10, and 8 to 30 years, respectively,
depending on clean sediment accumulation rates at each transect. PCB contamination at depth was
associated primarily with silt and silt/clay sediment layers. Little to no contamination was found in
sand layers. PCB congener compositions became increasingly dominated by lower chlorinated
71
-------
congeners with sediment depth and corresponding age, suggesting that dechorination is occurring to
some extent in the deeper and older sediments. This hypothesis was supported by a relative
accumulation of ortho and loss of meta and para chlorines in the PCBs analyzed in these sediments.
These minor changes in the degree of chlorination do not in any way imply a reduction in risk from
PCB contamination in these sediments.
Water and sediment samples were also collected from Lake Hartwell and a background site to
determine if PCBs in the sediments were bioavailable (by measuring fathead minnow liver
cytochrome P450IA1 gene expression) and/or if they were estrogenic (by measuring male fathead
minnow liver vitellogenin gene expression). Four water column samples (one each from the
background site and three transect locations) were tested on May 10, 2001 using 11-13 month old
adult male fathead minnows and 42-hr old fry. Fry were exposed to Lake Hartwell water for 48 hr
and the adult males for 24 hr. In addition to the Lake Hartwell samples, two positive controls and
two laboratory water controls were also tested. The positive controls were 50/50 mixtures of
Arochlors 1242 and 1258 at 10 and 100 ng/L. Nine sediments samples were tested with fathead
minnows. Two moderately hard laboratory water controls (without DMSO and with DMSO at 3.74
Tg/L) and one positive control (50/50 mixture of Arochlors 1242 and 1258 at 10 ng/L) were also
tested. Adult fish livers were necropsied after exposures, and RNA was isolated to determine if the
expression of P450IA1 and/or vitellogenin genes was increased. Fry were collected after each
exposure, and RNA was extracted and selectively amplified by PCR to also determine if these genes
were being expressed in embryos exposed to sediments. Preliminary results of this work will be
presented at the Workshop.
72
-------
Richard C. Brenner
Richard C. Brenner is an Environmental Engineer at the U.S. EPA National Risk Management
Research Laboratory (NRMRL) in Cincinnati, OH. He as a MS in Environmental Engineering and a
BS in civil Engineering from the University of Cincinnati. Mr. Brenner has many research interests
including: bioremediation and natural recovery of contaminated sediments, biotreatment and
chemical oxidation/biotreatment of contaminated soils, and fate of EDCs during wastewater
treatment. He has been the senior author or co-author on over 25 peer-reviewed journal articles. Mr.
Brenner has worked on over 35 conference proceedings articles and presentations.
James M. Lazorchak
Dr. James Lazorchak is a research Aquatic Ecologist and toxicologist at the US EPA Office of
Research and Development's Ecosystem Research Branch in Cincinnati, OH. As a Senior
Ecotoxicologist and manager of an AAALAC Certified Aquatic Research facility he is responsible
for exposure toxicity and molecular methods development and research. Dr. Lazorchak's latest
research is in the area of real-time biological monitoring using clams, daphnia, algae and fish to
detect episodic and long-term exposures to contaminants. He is also conducting research on the use
of gene expression in fathead minnows to detect estrogenic compounds, metals and chlorinated
persistent chemicals. He is the indicator lead for research and assessment methods for fish
contamination, water and sediment toxicity. Dr. Lazorchak is also the co-lead for zooplankton and
macroinvertebrates. Dr. Lazorchak is responsible for exposure design for genetic variation testing
with fathead minnows and amphipods. He has a Doctor of Philosophy in Ecotoxicology from the
University of Texas at Dallas. Dr. Lazorchak has a MS in Environmental Sciences also from the
University of Texas at Dallas, as well as a MS in Aquatic Ecology from Wright State University. He
has a BS in Zoology from Southeast Missouri State University. Dr. Lazorchak has been published in
numerous journals and US EPA publications including: Environmental Toxicology and Chemistry,
and Aquatic Toxicology.
73
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74
-------
PROGRAM FOR THE IDENTIFICATION AND REPLACEMENT OF ENDOCRINE
DISRUPTING CHEMICALS
Douglas Young Douglas Young
U.S. Environmental Protection Agency
Office of Research and Development
National Risk Management Research Laboratory
26 W. Martin Luther King Dr., MS-466
Cincinnati, OH 45268
(513) 569-7624
(513) 569-7111 fax
young.douglas@epa.gov
A computer software program is being developed to aid in the identification and replacement of
endocrine disrupting chemicals (EDC). This program will be comprised of two distinct areas of
research: identification of potential EDC and suggestions for replacing those potential EDC. This
identification portion of the program will be accomplished by constructing virtual 3-D representations of
potential EDC and then comparing these representations to a library of known EDC and to a library of
chemicals that have been identified not to be an EDC. These libraries will most likely focus around the
estrogen receptor sites since this is where a vast majority of the data is. This identification portion of the
research will be conducted external to the US EPA. The second phase of the program will focus on
suggesting replacements. This portion of the program will suggest a series of possible replacements for
potential EDC. The suggested replacements will be a chemical or a mixture of chemicals that will best
match the appropriate chemical and physical properties of the EDC. The portion of the program will be
configured similar to the software program PARIS II (Program for Assisting the Replacement of
Industrial Solvents), which was developed within the US EPA. This project to identify and replace EDC
is scheduled to begin in the Fall of 2001. This presentation will introduce a detailed plan for this project.
75
-------
Douglas Young
Dr. Young is a chemical engineer working at the U.S. Environmental Protection Agency's National
Risk Management Research Laboratory in Cincinnati, OH. There he leads the Simulation and
Process Design Team. Dr. Young's research is in the areas of environmental impact assessment as it
pertains to the chemical processing industry and the estimation of acute toxicity measurements by
structural relationships. Dr. Young was instrumental in the development and commercialization of
the generalized Waste Reduction (WAR) algorithm. Dr. Young received his Ph.D. from the
University of Arizona where his dissertation focused on the bioremediation of high-energy explosive
waste generated at the Los Alamos National Laboratory. He received his M.S. from The University
of Notre Dame and his B.S. from The University of Michigan. All of Dr. Young's degrees are in
chemical engineering.
76
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Using Bioassays to Evaluate the
Performance of
Risk Management Techniques
Carolyn Acheson, M. Christina Brinkman,
and Gregory Sayles
-------
Common Risk Management Assumptions
Risk - characterized by contaminants - Ignores
Incomplete removal or side products
Co-Contaminants
Matrix Effects
Treatment reduces toxicity - Ignores
Process Amendments
Other reactions
Matrix Changes
-------
Case Study 1
Remediation of PCB Contaminated Soil by
Solvent Extraction
Principals: Mark Meckes, John Meier, and Lina
Chang
More information - Meier, et al. 1997.
Environmental Toxicology and Chemistry, p. 928
-938.
-------
Case Study 1 - Chemical Analysis
*PCBs
*VOCs
4- SVOCs
Metals by TCLP
-------
Case Study 1 - Bioassays
Bioassay
Earthworm
Survival
Seed
Germination
Earthworm
Reproduction
Root
Elongation
Allium
Mitotic
Aberrations
Organism
E. fetida
L. terrestris
Oats and
Lettuce
E. fetida
Oats and
Lettuce
Allium
Exposure
Matrix
soil
soil
soil
soil
water
extract
Exposure
Period
14 days
5 days
3 weeks
5 days
24 hour
Endpoint
Survival
Survival
Survival, body mass,
number of cocoons,
cocoon hatchability
Growth
Mitotic index,
chromosomal
abnormalities
-------
Case Study 1 - Solvent Extraction
Solvent
J_
Soil + PCBs
Soil
Solvent + PCBs
-------
Case Study 1 - Results
100
80
Lettuce Seed 60
Germination
(%) 40
20
0
t Control
~
- After
w Treatment
_
-
-
~
Before
Treatment"
r
i-
-
0 50 100 150
PCB Concentration (mg/kg soil)
-------
Case Study 1 - Results
100
80
Lettuce Seed 60
Germination
40
20
0
t Control
™
" After Treatm
| 8.9g2-Propa
-
-
mt
nol/kg soil
-
~
Before
Treatment"
r
i-
-
0 50 100 150
PCB Concentration (mg/kg soil)
-------
Case Study 1 - Better RM
Solvent
Soil + PCBs
1
Solvent
PCBs
Water
I
Water +
2-Propanol
Soil
-------
Case Study 1 - Results 2
100
80
Lettuce Seed 60
Germination
40
20
0
t Control
JL Rinsed
" After Treatm
| 8.9g2-Propa
_
-
snt
nol/kg soil
-
~
Before
Treatment"
r
i-
-
0 50 100 150
PCB Concentration (mg/kg soil)
-------
Case Study 1 - Summary
Solvent extraction removed PCBs from soil
Process residues were as toxic as PCBs
Better RM - Add rinse step
Reduce PCB concentration
Reduce toxicity
-------
Case Study 2
Remediation of soil contaminated with wood
treating wastes by Soil Washing
Fluid - Ethanol-water mixture
Question - 2 or 3 Soil Washing stages?
Principals
Soil Washing - Richard Brenner, Makram Suidan,
George Sorial, Amid Khodadoust, Karen Koran, and
Gregory Wilson
Ecotoxicity Evaluation - Carolyn Acheson, Jennifer
Mansfield , Yonggui Shan, and Margaret Kupferle
-------
Case Study 2
Chemical Analysis
PCP
Hydrocarbons - alkanes and PAHs
Bioassays
Earthworm Survival
Seed Germination and Root Elongation in Lettuce and
Oats
-------
Case Study 2 - Soil Washing
Contaminated
Soil
Wash 1 Wash 2
H2O
I
H2O
2 - stage
Treated
Soil
Wash 1 Wash 2 Wash 3
H2O
H2O
3-stage
Treated
Soil
-------
Case Study 2 - Results
Chemical
Concentration (mg/kg dry soil)
Untreated
2-stage
3-stage
PCP
950 ±51
31± 1
9± 1
Alkanes
1761 ±46
130 ± 17
59 ±12
PAHs
494 ± 20
12 ±2
<3
-------
Case Study 2 - Results
Earthworm
Survival
20 40 60 80
Test Soil Concentration
(% test soil in total soil)
Untreated
2-stage
3-stage
100
-------
Case Study 2 - Results
untreated
2-stage
• 3-stage
20 40 60 80
Test Soil Concentration
(% test soil in total soil)
100
-------
Case Study 2 - Results
Bioassay
Untreated
2-stage
3-stage
LC 50 (% test soil in total soil)
Earthworm
Survival
Lettuce Seed
Germination
4.0
5.1
70.7
>100
35.0
(26.3, 43.4)
89.4
(74.4, >100)
EC50 (% test soil in total soil)
Lettuce Root
Elongation
Oat Root
Elongation
*
12.1
(0, 23)
17.9
(7.6, 23.0)
49.8*
(44.7, 55.7)
16.2
(12.6, 20.0)
44.6*
(35.1,61.4)
* Response in reference toxicant or negative controls were not in expected range
-------
Case Study 2 - Summary
Soil Washing was effective
Chemistry
+ PCP, Alkanes, and PAHs removed
+ 3- stage process most effective
Bioassays
+ Earthworms and plants show reduced toxicity in
treated soils.
+ 2-stage process most ecologically hospitable
Likely that soil washing alters other aspects of soil
-------
Risk Management of EDCs
Uncertainties of EDCs
Unknown endocrine activity of degradation products
Unknown effectiveness of treatments in reducing
endocrine activity
Concurrent chemical and biological measures of
effectiveness recommended
-------
EDC Bioassays - Considerations
EDCs of concern in NRMRL projects
Concentrations
Environmental Matrices
Data Quality - Reproducibility and Reliability
Practicality - Cost and Ease of Use
Recommendations of Others
Adaptability to RM projects
-------
EDCs of Concern in NRMRL projects
Alkylphenols
Chlorinated Dioxins and Furans
Estrogens, biogenic and pharmaceutical
All are estrogenic;
some have thyroid and developmental effects
-------
EDC Bioassays - Considerations
Environmental Matrices in NRMRL projects
Water
Solids - soils, sediments, and biosolids
Concentrations
Water - as low as ng 17 p-estradiol/L
solids - levels vary
-------
EDC Bioassays - Considerations
Data quality Evaluated by
Practicality ^ ^eer reyiewed literature
EDSTAC report
A j . 1 -iv
Adaptability
+ ORD colleague
Sensitivity recommendations
-------
Types of EDC Bioassays Considered
Sediment/Aquatic Invertebrate tests
Terrestrial Invertebrate tests
In vitro tests
Fish Vitellogenin mRNA assay through
cooperation with MERB/NERL
-------
Sediment/Aquatic Invertebrate tests
Advantages
Commonly studied aquatic organisms
Many endpoint options
Disadvantages
Mechanism of action - interference
with molting controlled by steroid
hormones, ecdysteroids
Require substantial lab equipment
Test duration - about 1 month
From www.AquacultureStore.com
-------
Terrestrial Invertebrate tests
Imposex Occurrence - measure of androgenicity
Earthworm Reproduction
methods exist
endpoints such as number of cocoons and number of
hatchlings per cocoon
endpoints are not directly related to endocrine function
unknown sensitivity to EDCs of concern
-------
In Vitro Assays
Mammalian cells: E-Screen and MVLN
organism - immortal mammalian cell (MCF-7)
endpoint - proliferation or luciferase production
MVLN - recommended by EDSTAC
Yeast Estrogen Screening Assay (YES)
evaluated by EDSTAC
commonly used in peer reviewed literature
not recommended for chlorinated pesticides
Both - reported sensitivity at low concentrations
-------
EDC Bioassay Selected for Adaption -
YES Assay
o
I
Estrogen
Estrogen
Receptor
Activated
Receptor
I > Reporter
Protein
O • CPRG
lacZ
From Routedge and Sumpter, 1996. Environ. Tox and Chem. 15: 241- 248
-------
NRMRL Sponsored EDC RM Projects
Using Bioassays
Project
Evaluation of Drinking Water Treatment
Techniques for EDC Removal
Potential of CAFOs to Contribute
Estrogens to the Environment
Investigations of Sorption and Transport of
Hormones and Animal Pharmaceuticals
Evaluating the Fate of EDCs During
Wastewater Treatment
EDCs from Combustion and Vehicular
Emissions
Natural Recovery of PCBs in Sediments
EDC
Steroid Hormones
Alkyl Phenols
Estrogens
Estrogens
Steroid Hormones
Alkyl Phenols
PCBs
Dioxins/Furans
PCBs
Prinicpal
Investigator
Kathleen
Schenck
Steven
Hutchins
Suresh Rao
Carl Enfield
Paul
McCauley
Brian Gullett
Richard
Brenner
Bioassay
MVLN Assay
FETAX
XTRA
YES Assay
YES Assay
Vitellogenin
mRNA Assay
Vitellogenin
mRNA Assay
Vitellogenin
mRNA Assay
-------
Using Bioassays in a
Hypothetical EDC RM Project
EDC of Concern - Phthalates
commonly used as a plasticizer in many household
products (including food containers)
suspected to cause alterations in human sexual
development
RM Project - Find a replacement plasticizer for
phthalates
-------
Hypothetical EDC RM Project - Phthalate
Replacement
Use computer models to find substances with
appropriate chemical and physical properties
Lab Testing of leading candidates
chemical and physical testing to determine if acceptable
substitute
bioassays to evaluate biological activity
Look at production processes
Are production by-products likely to cause problems?
Test bulk chemical - chemical, physical, biological
properties
-------
Acknowledgements
Jennifer Mansfield, IT Corporation
Kathleen Schenck, U.S. EPA
Andrew Avel, U.S. EPA
-------
Natural Recovery of PCB-Contaminated
Sediments at the Sangamo-Weston/
Twelvemile Creek/Lake Hartwell
Superfund Site
Richard C. Brenner1, Victor S. Magar2, Glenn Johnson4,
Gregory S. Durell3, Eric A. Crecelius5, James E. Abbott2,
Jennifer A. Ickes2, and Carole S. McCarthy3
1U.S. EPA, NRMRL, Cincinnati, OH 2Battelle, Columbus, OH
3Battelle, Duxbury, MA "University of Utah, Salt Lake City, UT
5Battelle, Sequim, WA
-------
Magnitude of Sediment Contamination
U.S. EPA estimates that 6 to 12% of the sediment
underlying the nation's surface water is potentially
contaminated (Long etal., 1996; U.S. EPA, 1997)
720,000,000 to 1,440,000,000 yd3 of contaminated
sediment reside in the upper 5 cm (U.S. EPA, 1997)
Baltelle
-------
Monitored Natural Recovery
Monitored Natural Recovery (MNR) of sediments is a
remedial option that relies on natural environmental
processes to permanently reduce risk, and which
includes careful assessment, modeling, and monitoring to
ensure success (as defined by the RTDF)
Natural processes most often associated with MNR:
• Sediment containment through natural capping
• Requires net depositional areas
• Contaminant weathering
• Biological Processes
• Physical/chemical processes
• Contaminant sorption/sequestration
Baltelle 3
-------
Natural Capping and Burial
Largest contributor to natural recovery
• Natural barrier to the aquatic environment
• Reduces exposure of water column with contaminated
surface sediments
• Reduces upward contaminant diffusion and advection
Reduces contaminant bioturbation and transport into the
food chain
Potential for resuspension
• Storm events
• Construction/industrial activities
Baltelle
-------
Natural Recovery Program Objective
• Investigate natural recovery of contaminated
sediments at two sites
• PCB-contaminated sediments
Sangamo-Weston/Twelvemile Creek/
Lake Hartwell Superfund Site (Pickens County, SC)
• PAH-contaminated sediments
Wyckoff/Eagle Harbor Superfund Site
(Bainbridge Island, WA)
• Develop field evaluation techniques
• Use a snapshot approach
ERA Baltelle
-------
Lake Hartwell Site, South Carolina
&EPA
Documented history of
contaminated sediments
• Capacitor Manufacturing
(1955-1978) - estimated 400,000
tons PCBs discharged
• Single primary PCB source
(Aroclors 1016,1242, and 1254)
Natural Recovery selected to
restore Lake Hartwell sediments
(EPA ROD, R04-94/178)
Terrestrial PCB contamination
has been removed/contained
Battelle
-------
Lake Hartwell Site Map
Denotes Sample
Transect Location
• Sediment cores
collected at
centerline of 10
transects - three
cores/transect
• Transects were
established by
EPA Region 4
• Extruded
samples after
coring
• Transect
Locations
T16, W7, Q, P,
O, N, L, J, I, T6
Battelle
-------
Upgradient Sample Collection at Lake Hartwell
Collected sediment cores
Extruded cores into
5-cm segments
&EPA
Battelle *
-------
Downgradient Sample Collection at Lake Hartwell
xvEPA
Battelle
-------
Lake Hartwell Results
Three sets of results will be discussed:
Vertical and horizontal PCB distribution
Sediment age dating [using lead isotope
(210Pb) analyses] and sedimentation rates
PCB homologue and congener distribution
analyses
-------
Vertical t-PCB Concentration Profile
Transect Q (Upgradient)
^ 0-10
<£> 15-25
^C
+± 30-40
Q.
Q 40 -50
'c 55-65
O_
E65-75
0? 83-93
CO
-------
Vertical t-PCB Concentration Profile
Transect T16 (Upgradient)
*£ 0-10
^ 20-30
£ 40-50
g 55-65
'g 77-87
£ 97-107
m
0 110-117
(/)
m 117-124
O
O 128-138
O
138-148
=]
I
69 err
41 err
OA rrt
24 err
0 20,000 40,000 60,000 80,000 100,000
Concentration (M9/kg) dry weight
-
Sand
Silt/Sand
Silt
Sand
-------
Vertical t-PCB Concentration Profile
Transect L (Downgradient)
82.5
1999
1997
1994
1991
1988
1985
1981
1977
1973
1969
1965
1960
1956
1951
1944
1935
1926
10,000 20,000 30,000 40,000 50,000
Concentration (pg/kg) dry weight
-V
•:•
I
Silt
1
3?
•
"3
Baiteife"
-------
210Pb and 137Cs Results
Assumptions for uniform flux of 210Pb onto surface
sediments
• Uniform grain size throughout sediment profile
• Constant historical sedimentation rate
Assumptions met in downgradient Cores 0, N, L, I, and T6
Upgradient cores and Core J could not be dated
Very low or variable 137Cs concentrations meant
137Cs could not be used to date sediments
Baffelfe"
-------
Sediment Accumulation Rates
25.0
20.0
15.0
+- JS 10.0 -•
0
CO
o
15
31
o <*,
5.0
2000
Core
Sedimentation Rate
(g/cm2-yr)
O
N
L
I
T6
3.45
1.70
2.23
2.93
0.75
Average
2.85 + 1.79
1980
1960
Date
1940
1920
Baffelle"
-------
Estimated Additional Sedimentation (cm)
Needed to Achieve Sediment Cleanup Goals
Core
O
N
L
1
T6
Avg.
1 mg/kg 0.4 mg/kg 0.05 mg/kg
t-PCB t-PCB t-PCB
2.8
0
2.7
0
0
1.2 ±1.7
16
7.8
11
11
3.5
10±4.7
45
29
31
42
13
32 ±13
• 1 mg/kg: ROD surface sediment cleanup goal (EPA, 1994)
• 0.4 mg/kg: Mean site-specific sediment quality criteria (EPA, 1994)
• 0.05 mg/kg: NOAA effects range-low (EPA, 1994)
&EPA
-------
Estimated Additional Time (yr)
Needed to Achieve Sediment Cleanup Goals
Core
O
N
L
1
T6
Range
1 mg/kg
t-PCB
1 -3
3-5
1 -5
0.4 mg/kg
t-PCB
8-10
5-10
5-7
2-5
2-5
2-10
0.05 mg/kg
t-PCB
>28
25-30
15-20
10-15
10-15
10-30
• 1 mg/kg: ROD surface sediment cleanup goal (EPA, 1994)
• 0.4 mg/kg: Mean site-specific sediment quality criteria (EPA, 1994)
• 0.05 mg/kg: NOAA effects range-low (EPA, 1994)
&EPA Baltelle"
-------
Core L Homologue Plots
CM through CI-3
00 co
O -i-
Q. O
i08
OB\
CNI
O ,_-
<5 -L
Q. O
1O1O1O1O1O1O1O1O1O1O1O1O1O1O1O1O1O
80%
70%
60% (D
50%^ S
40%
3 2,
30%^
20%
10%
0%
Q
Depth (cm)
-------
Core L Homologue Plots
CI-4 through CM 0
co
o
o_
0)
e
0)
0_
80%
70%
60%
50% -
40% -
30% -
20%
10%
0%
-*- CI-4
-•- CI-6
-A- CI-5
— Sum
IOIOIOIOIOIOIOIOIOIOIOIOIOIOIOIOIO
Depth (cm)
2.0%
CO | 1.6%
Oco
CI-8 Sum
CI-10
IOIOIOIOIOIOIOIOIOIOIOIOIOIOIOIOIO
• 0.10%
• 0.08%^ J
0.06%£ §
3 «-"
Q. O
• 0.04% 0 2
• 0.02%o O
0.00%
Depth (cm)
-------
PCB Congener Distribution
in Surface and Deep Sediments
CQ HO
O '*
O- o
O)
£
O)
Q_
CQ
O
Q_
O)
£
O)
Q_
CQ
O
Q_
O)
£
O)
Q_
0
12
9
6
3
0
12
9
6
3
0
-3
-6
Core L, Segment 1, 0-5 cm depth interval
Total PCB = 1.58mg/kg
TL
Segment 8, 35-40 cm depth interval
Total PCB = 48.7 mg/kg
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PCB Congener
Baffelfe"
-------
Summary and Conclusions
Highest t-PCB concentrations associated with silt/clay layers
Decreasing surface t-PCBs, at or approaching 1.0 mg/kg
(Max. surface PCB concentration = 1.58 mg/kg at Transect L)
Time to achieve surface sediment concentrations
• 0 to 5 yr to achieve 1.0 mg/kg
• 2 to 10 yr to achieve 0.4 mg/kg
• 10 to 30 yr to achieve 0.05 mg/kg
Homologue shifts from higher to lower chlorinated congeners
• CI4/CI5/CI6 congeners reduced from 80% to 20% t-PCB with depth and time
• CI1/CI2/CI3 congeners increased from 20% to 80% t-PCB with depth and time
Significant accumulation of ortho chlorinated congeners
&EPA
-------
Effectiveness of Natural Recovery Approach
Conclusions
High resolution PCB chromatography (107 congeners eluted) used
to characterize vertical PCB concentration profiles and PCB
dechlorination patterns with sediment depth and age
Sediment isotope analyses provided an effective means of
calculating sedimentation rates and surface sediment recovery
rates
Future Work
Evaluate relationship of decreasing surface sediment contamination
with benthic animals and fish toxicity
Assess long-term stability of natural recovery progress at this site
&EPA
-------
Removal of Endocrine
Disrupting Chemicals from
Drinking Water using
Granular Activated Carbon
John L. Cicmanec, DVM, MS
Thomas F. Speth, PhD
Kathleen M. Schenck, BS, MS
National Risk Management Research
Laboratory, Office of Research and
Development
-------
Untied Slates Office at Research and EPA/62SR-OIM015
Environmental Protection Development March 2001
Agency Washington. DC 20*60 www/epa^goWORD
Removal of Endocrine
Disruptor Chemicals
Using Drinking Water
Treatment Processes
-------
Selected Endocrine Disrupting
Chemicals
Phthalates
Alkylphenols and AP ethoxylates
Ethinylestradiol
Methoxychlor
Endosulfan
razine
RGBs, Dioxins, and Dibenzofurans
Bisphenol A
-------
Evidence for Endocrine Activity
^
Compound
DDT
PCBs
Dioxins
Alkylphenols
Ethinvlestradiol
Lab
X
X
X
X
1 X
Eco
X
X
X
X
X
m^m
Hum
X
X
X
-------
Evidence for Endocrine Activity
Compound Lab
Methoxychlor X
Endosulfan X
DEP X
DEHP X
Bisphenol A X
Eco
Human
-------
Basis for Selection of EDCs
Ecological
DDT
PCBs
egg shell thinning Nelson, 1978
gulls-sex ratios Schreiber,1970
Alkylphenols vitellogenin
Jobling,1995
Dioxins female-female pairs Hunt, 1984
-------
Basis for Selection of EDCs
Human Data
RGBs reduced birth weights
Patandin/98, Taylor, '78
Dioxins reduced number of male births
Seveso, Italy
T delayed mental development from
in utero exposure, Rogan, 1978
111
-------
Reproductive Endocrine effects
of PCBs (Direct evidence)
Arochlor 1254 in Rhesus monkeys-altered
progesterone levels at 5 M9/kg and 80
Mg/kg (Arnold, 1995)
Arochlor 1016 in Rhesus monkeys-
decreased estrogen levels at 28
(Barsotti, 1976)
-------
Reproductive Endocrine effects
of PCBs (Indirect evidence)
Arochlor 1248-100 M9/k9 in Rhesus
monkeys 5/8 live births initial
gestation as well as residual effects
, & months later
-------
"On the Horizon"
Coxsackievirus - juvenile diabetes
Helicobacter pylori - thyroid disorders
Sulfonamide antibiotics - thyroid effects
Trenbolone acetate
Melengestrol acetate
-------
Human Exposure Levels vs
Endocrine Effects in Lab
J ^Animals
p,p' DDE
Human fetus, first trimester
Rat fetus, sex differentiation
Dioxin
Human body burden
Rat fetus, sex differentiation
MEHP
Child on dialysis
Adult Rat (1 g/kg 14 d)
11 ppm
1-20 ppm
13ppt
5-40 ppt
2.4-15ug/ml
48-152ug/ml
-------
Environmentally Relevant
Doses
Nonylphenol
1000 ug/L. Rhine River
1000 ug/L. Savannah River
75 ug/L. Swedish Rivers
6-160 ug/L. Kansas River
-------
Occurrence of Nonylphenol
The concentration of NP in sewage
effluent decreased from 75 ug/L. to
9 ug/L. in one year in Sweden
Found in 24% of water samples in
U.S.
-------
Conventional Drinking Water
Treatment
Coagulation
Flocculation
Sedimentation
Filtration
Chemical disinfection
-------
The Capacity of Conventional
Water Treatment Processes to
J^ Remove EDCs
87% of methoxychlor removed by
coagulation with pH adjustment
98% of DDT is removed by coagulation
35% of PCBs removed by coagulation &
filtration
Removal of lindane, aldrin, dieldrin &
pa rath ion varied from 10-60%
-------
Specialized Water Treatment
'^ Processes (Organic removal)
Air Stripping
Activated carbon
(Granular and Powdered)
Membrane filtration
nanofiltration, reverse osmosis
Ultraviolet Light
-------
Powdered Activated
Carbon
-------
-------
Present Usage of GAC and
PAC is used in 48% of surface water
treatment plants and 12% of ground water
plants
GAC is used in 12% of surface water treatment
plants and 5% of ground water treatment
plants
-------
Factors for PAC/GAC Treatment
Contaminant adsorbability
Concentration
Multicomponent adsorption
Adsorption kinetics
Temperature
Carbon dose
Contact time
-------
Freundlich equation
Q=KxCn
=Equilibrium capacity of carbon
= Equilibrium capacity of liquid-
phase concentration of target compound
= Freundlich coefficients
-------
Isotherm Constants for
Selected EDCs
DDT
Endosulfan
Diethylphthalate
DEHP
PCB-1221
PCB-1254
PCB-1260
Nonylohenol
10,499
2,548
17,037
8,308
1,922
1,050**
3,700**
19,400
Note: Values above 200 are economically feasible
-------
Isotherm Prediction (Speth and
Adams, 1993)
CJ/MW
W0*exp(ps*B*R*T*ln(1000
l/n
K = Freundlich K in
ps = density of pesticide (g/mL)
W0 = 0.808 mL/g
B = -0.075 mL/cal
R = ideal gas constant (1.987 cal/gmol/°K)
T = temperature (Kelvin)
Cs = pesticide solubility (mg/L) *
MW = molecular weight of pesticide*
Prediction can be off by an order of magnitude!
-------
-------
-------
-------
. i
•
.
'.
m
-------
-------
Granular Activated Carbon
Treatment at CCW
Present capacity is 200 million
gallons/day
Cost to install granular activated
carbon system was $60 million
Present cost to consumers is 27
cents/1000 gallons vs 25
cents/1000 gallons before
-------
Conclusions
Removal of EDCs by conventional treatment
processess is variable
GAC & PAC can remove most hydrophobic
compounds such as many of the suspected
EDCs
Powdered carbon can remove most EDCs
inexpensively and is particularly useful for
seasonal application
-------
ebsite
http:/www.epa.gov/ttbnrmrl
-------
Introduction to EDCs and their
Effects on Humans
Ralph L. Cooper, Chief
Endocrinology Branch
Reproductive Toxicology Division
National Health and Environmental
Effects Research Laboratory
-------
Overview
Introduction to Endocrine System
Research Findings
• Lines of Evidence, Outcomes, Modes,
Chemicals
Endocrine Disruptor Screening
Program
• Mammalian Tests
-------
Pineal
othalamus
Pituitary
Thyroid
Parathyroids
Thy mus
Adrenals
Pancreas
Ovary
i
-------
Hormones of Male Reproduction
Hypothalamus
Pituitary
Hypothalamus
Testosterone
ituitary
FSH
Testes
Epididymis
-------
Male Reproductive System
Accessory Organs
Seminal Vesicle
Ejaculatory duct
Prostate gland
Bulbourethral gland
Vas deferens
Epididymis
Scrotum
Denis
-------
Hormones of Female Reproduction
Hypothalamus
Pituitar
Hypothalamus
lituitary
Estrogen
Progesterone
Ovary
-------
Female Reproductive System
•Ovary
•Uterine Tube
Uterus
•Cervix
Vagina
Ovarian preovulatory follicle
-------
Thyroid Hormone Regulation
HvDothalarr
Pituitar
Hypothalamus
Parathyroid gland
Thyroid
gland -
T3 and T4
-------
Communication
Receptor
Cell-to-Cell and
Organ to Organ
Secreting cell
Target cell
Not a target cell (no receptors)
Target eel I
Secreting cell
Blood vessel
Target cell
-------
Classical Pathway for Estrogenic
Activity Direct Gene Induction
ERE
' M.t i
ER
ER
^
E
ER
^^H
E
Coactivators,
Nucleus
Transcription
mRNA
mRNA
Tra
New
protein"
Physiological
response
-------
1.
Outside
of cell
Inside
of cell
Membrane Receptors
(protein hormones)
Hormone Plasma
membrane
Receptor molecule
Inactive
G-protein
Membrane
Nucleus
2.
Receptor binds hormone
Hormone-receptor
complex activates
G-protein
Inactive
adenylate
cyclase
Active G-protein
activates adenylate
cyclase
Active
adenylate
1 cyclase
cAMP
1
Other reactions
-------
Steroidogenesis
cholesterol
± cyp P450-SCC
17-alpha-hydroxylase
^^
iregnenolone
3-beta-hydroxysteroid
dehydrogenase
hydroxypregnenolone
17,20-lyase
progesterone
T7-alpha-
dihydroepiandrosterone
17-hydroxyprogesterone
3-beta-hydroxysteroid
dehydrogenase
17,20-lyase
androstenedione
aromatase
17-beta-HSD
testosterone
aromatase
estrone
estradiol
FSH
-------
Summary
Endocrine system is a complex network of
coordinated regulatory changes.
Homeostatsis
Endocrine system provides a variety of targets
for enviornmental chemicals
• Receptors (nuclear and membrane)
' Synthesis
Clearance
-------
Endocrine Disrupters and
Human Health
Definition
• An exogenous substance that alters
function(s) of the endocrine system
and consequently causes adverse
health effects in an intact organism,
or its progeny, or (sub)populations.
IPCS, 1998 (Global Assessment)
-------
Modes of Action
Homologous Mechanisms
Receptor binding
. ER, AR, AhR, GR, GABA
• Agonists and Antagonists
Enzyme Inhibition
• Steroidogenesis, thyroid peroxidase
Enzyme induction
• CYP450s, thyroxine conjugation
Signal transduction pathways
• Phosphatases/kinases, transcription factors
-------
Lines of Evidence
Laboratory based toxicology studies
Anti-androgens, estrogens, chlorotriazines
Field Studies
Lake Ontario Lake Trout and dioxin (TEQs)
Status and Trend Studies
Population status, tumor incidence, contaminant
loads
Epidemiology
PCBs and developmental neurotoxicity
Developmental Susceptibility
Each with Strengths and Limitations
Cost, Sensitivity, Predictiveness
-------
Experimental Evidence
Effects on Developing Reproductive Tract
i Estrogens (DES, Methoxychlor, Bisphenol A)
• Anti-Androgens (Vinclozolin, DDE, Phthalates)
• Ah Receptor Agonists (dioxin, PCBs)
• Steroid Synthesis Inhibitors
Developmental Neurotoxicity
i OH- PCBs (hypothyroidism),
• Estrogens (masculinization)
• Polybrominated diphenylethers (hypothyroidism)
• Thyroid Gland
• Leydig Cell Hyperplasia
• Mammary Tumors (Atrazine)
-------
Phenotypes in Exposed Offspring
Estrogens
• Females » Males
• Brain masculinization
• Altered Puberty (>f, » Females
• Feminized genitalia, Hypospadias, Nipple
Retention, Organ growth
Ah Agonists
• Male > Females
• Reduced sperm release, Vaginal threads,
Accelerated eye opening
-------
Reported Human Health Effects
• Breast Cancer
tir
Shortened Lactation
in Male Reproductive Health
• malformations, semen quality, cancer
Altered Immune Function
Developmental Learning Disabilities
-------
Declining Reproductive Health
META ANALYSIS OF SPERM COUNT
Sperm Counts
150
100-
50
O
o o
[ \^ I
I
I
1930 1940 1950
1960
Year
1970 1980 1990
Carlsenetal, 1995 BMJ 305:609
-------
Increased Malformations
Hypospadias
45
4O
-------
PCBs and Human Development
Rice-Oil Poisonings
• Japan, '68; Taiwan,
79
• Yusho, Yu-cheng
• 4000 exposed
. PCBs: 1 g, 40 ppm
. PCDFs: 38 mg, 15
ppt
• Perinatal Deaths
• Low Birth Weights
i Hyperpigmentation
• Gum hypertrophy
• Delayed Maturation
• Cognitive Deficits
Environmental Exposures
• Four Cohorts
. Fish Eaters (LM, EU)
• Nursing Mothers (NC)
. Gen. Population (NL, EU)
• Motor Deficits
• hypotonia
• Memory Deficits
• Immune Impairments
• Thyroid Effects
. (TSH, T4)
• Consistent??
• Transient??
Prenatal Critical Period
-------
Classes of EDCs
Effluents
Flame Retardants
Fungicides
Herbicides
Insecticides
Metals
Pharmaceuticals
Phenols
Plasticizers
Polyaromatic
Hydrocarbons
Soy Products
Surfactants
BKME, STW
PBDEs
Vinclozolin
Atrazine
Methoxychlor
Tributyltin
Ethynyl Estradiol
Bisphenol A
Phthalates
PCBs, dioxin
Genistein
Alkylphenol
Ethoxylates
-------
Major EDC Uncertainties
• Exposure-Outcome Linkages
• Latency
• Persistent vs. non-persistent contaminants
• Fate and transport
• What effects are occurring in humans?
• Comparative toxicology
• Sequence homology, binding, action
• Dose-response relationships
• Shape, monotonicity
Interaction with "endogenous" diseases
Testing protocols
-------
Major Uncertainties (Cont'd)
Chemical diversity
• ~100; Structures and potency; phytoestrogens
• What will EDSP tell us?
Multiple mechanisms of action
• >1 receptor, co-factors and co-repressors
• Dissimilar modes and similar phenotypes
• Polymorphisms/Environmental Genome
Project
Cumulative exposures and effects
Do EDCs need special consideration in
risk assessment?
-------
Summary
Is human health at risk?
• Data is present in animals
• Potential is present in humans
• Evidence in humans is controversial
• Dose (exposure)
> Time (Sensitive periods)
-------
FOOD QUALITY PROTECTION ACT
1996
Endocrine Disrupter Screening Program
• EPA Must
• Screen pesticides for estrogenic effects that
may affect human health
• Develop a screening program
• Report to Congress by August 2000
priority setting
• validation of Tier 1 and Tier 2 assays
-------
Framework
Initial Sorting
Priority Setting
Screening (Tier 1)
• Identifies substances for further testing
Testing (Tier 2)
• Identifies adverse effects and
establishes dose-response relationship
for hazard assessment
-------
PROPOSED SCREENING BATTERY
(Tierl)
• In vitro Screens
• ER Binding / Reporter Gene Assay*
• AR Binding / Reporter Gene Assay*
• Steroidogenesis Assay with minced testis
• In vivo Screens
• Rodent 3-day Uterotrophic Assay (sc)
• Rodent 20-day Pubertal Female Assay with
Thyroid
• Rodent 5-7 day Hershberger Assay
• Frog Metamorphosis Assay
• Fish Reproduction Screening Assay
These assays are in the HTPS
-------
ALTERNATE SCREENING ASSAYS
Rodent 20-day Pubertal Male Assay with
Thyroid
Placenta! aromatase
Rodent in utero through Lactation Assay
-------
Endocrine Disrupter Screening Program
Tier 1 assays have the necessary breadth and depth to
detect all currently known chemicals that may affect the
endocrine, androgen and thyroid systems. Therefore, after
having gone through the Tier 1 Screening battery, a
chemical will be designated as having either:
• the potential for estrogen, androgen or thyroid activity,
which will require further analysis
• in Tier 2 tests to verify and evaluate that potential; or
low or no potential for estrogen, androgen or thyroid
L activity, which will allow the chemical to be put on
"Hold."
-------
Progress on Mammalian Tests
Uterotrophic
Hershberger
Male and Female Pubertal
In utero/lactational
-------
Summary
Endocrine Disrupter Screen ing Program
• Methodical evaluation of chemicals
• Tier 1 test will identify most EDCs in vitro
and in vivo
• Tierl identifies mode of action
• Tier 2 test will evaluate potency and risk to
human
-------
EPA's Endocrin
Screenin
nf Im
HJJl
EPA
Elaine Z. Francis, Ph.D.
National Program Director for
Endocrine Disruptors Research
Workshop on Effective Risk Management of
Endocrine Disrupting Chemicals
January 29, 2001
-------
Regulations and Recommendations
. FQPA and SDWAA
. EDSTAC
Implementation
• Setting Priorities
• Proposed Screening and Testing
• Standardization and Validation
Research
• Supporting EPA's screening & testing program
• Overview of other EPA research
Summary
-------
-------
i
.I
, •.
I I L
Food Quality Protection Ac
I ' I ,
i I
Safe Drinking Water Act
i j
i i
I f , j
-------
A
Don
kv
•
andates
»
EPA must:
me e
iissi j qwp&fgrssiro c
^^_-__~^ ^
that may affect human health
Develop a screening and testing program
by Aucust 1993
.. - -
•program by August 1909
Report to Congress by August 2000
' * •" *' **
\
-------
A - SDWA
tjonary Authority
EPA can
j
\
cnsrrjj
e screening and testing of:
active or inert ingredient
:l that has an effect cumulativ
ct of a pesticide
Itfin Icing
water source contaminants
PTTprp
^f J J v^-wi.-
•-
5
L^*>, M
-------
r
and Testing
i
IJJ.
Chartered Oct 16, 1996 (www.epa.gov/scipoly/oscpendo)
39 members representing broad constituencies
Advise EPA on:
• A strategy for selecting and prioritizing chemicals for
screening and testing
• A process for identifying new and existing screening
assays
• A set of available screens for early application
• What tests and when to test beyond screening
• A process for identifying mechanisms for validation of
screens and tests
-------
Scope of Screenm
ogr
J7.
Based on current science and existing
EPA testing authorities, EDSTAC did not
limit the screening program to FQPA
and SDWA
• Estrogen, Androgen and Thyroid hormones
• Human health and ecological effects
• Expanded universe of chemicals and mixtures
-------
Chemicals Under EPA's Purview
• 900 pesticide active ingredients
. 2500 "inerts" in 20,500 products
• 75,500 industrial chemicals on the TSCA inventory
• Environmental contaminants (?)
Chemicals Under Others' Purview
• Cosmetics
• Food additives
• Nutritional supplements
-------
Initial Sorting
Priority Setting
Screening (Tier 1)
• Identifies substances for further testing
Testing (Tier 2)
• Identifies adverse effects and establishes
dose-response relationship for hazard
assessment
-------
Restrict consideration of Phase I chemicals to
pesticide active ingredients (-950) and chemicals
which are both "inert" formulation ingredients and
HPV commercial (-600) chemicals
Meets statutory mandate for screening pesticides
Robust statutory authority for obtaining data
Reasonable size group of chemical candidates to
focus on (1500-1600)
-------
Current EDSP schematic for Phase I
(estimated 87,000)
'Sticide active
HPV-lnerts
Pesticide actives
lower priority HPV-lnerts
higher priority HPV-inerts
insufficient data:
eed Tier 1 screen
Need Tier
testing
Adequal
data
assessm
-------
Goal: Set priorities for chemicals
according to potential exposure and
potential endocrine effects
How:
• Develop Priority Setting Database
• Group chemicals based on common effects
and exposure information
• Compare chemicals within groups and
establish priorities within groups
• Pick highest priority chemicals in each
group
-------
Default Scenario
File E_dit What If Chemicals
H.glp
Specially Targeted Priorities
I Exposure-Related Information
El-Human Biological Monitoring Data
Ep-Ecological Biological Monitoring Data
Ep- Chemicals in Food and Drinking Water
Ep- Chemicals in Consumer/Cosmetic Products
Ep- Occupational Exposure Chemicals
Ep Surface Water Monitoring Data
Ep- Indoor Air Monitoring Data
Ep-Sediments/Soil Monitoring Data
Ep- Persistence
Ep- Bioconcentration
Ep-Environmental Releases
Ep Production/Import Volumes
Ep - Superfund Data
EE1- Outdoor Air Monitoring Data
Effects-Related Information
Ep-Epidemiological and Clinical Data
Ep- Reproductive/Developmental Toxicity
Ep- Carcinogenicity
Ep-Subchronic Toxicity
Ep-Ecotoxicity
EE1 -Quantitative Structure Activity Relationships
Combined Exposure- and Effects-Related Informal
All Compartments
Exposure-Related Information
Category Statistics |
>
Compartment
Persistence
Sediments/Soil Mor
Outdoor Air Monitor
Superfund Data
Chemicals in Food ;
Occupational Expo:
Human Biological Iv
Environmental Rele
Bioconcentration
Production/Import \
Chemicals in Consu
Indoor Air Monitorin
Ecological Biologic;
Surface Water Mor
Number of
Chemicals
103,435
247
363
419
3,248
13,452
167
442
1 03,433
6,259
3,162
278
191
226
Weight
0
1
1
1
1
1
1
0
0
0
1
1
1
1
Relative
Weight
0
7
7
7
7
7
7
0
0
0
7
7
7
7
Pief
Level
50
51
52
53
55
57
59
61
62
63
64
65
66
67
Rankin
Techni
Weighte
Weighte
Weighte
Weighte
Weighte
Weighte
Weighte
Weighte
Weighte
Weighte
Weighte
Weighte
Weighte
Advance
*\ 1
EDPSD v.2 Exposure Data Screen
-------
Default Scenario
File £dit What If Chemicals Fieports Help
B|g| liN *1|
Speciallv Targeted Priorities
Exposure-Related Information
iEffects-Related Information
H Epidemiological and Clinical Data
ED- Reproductive/Developmental Toxicity
S- Carcinogenicity
S-Subchronic Toxicitv
EJ-Ecotoxicity
S- Quantitative Structure Activity Relationships
Combined Exposure- and Effects-Related Informal
All Compartments
Effects-Related Information
Category Statistics |
^
Compailmenl Number of Wej h
Chemicals
Ecotoxicity 308 1
Carcinogenicity 65 1
Reproductive/Deve 480 1
Epidemiological anc 80 1
Relative Pref Rankin
Weight Level Techni
7 54 Weighte
7 56 Weighte
7 58 Weighte
7 60 Advance
EDPSD v.2 Effects Data Screen
-------
or priority Sett
PROPOSED SOLUTION # 1
• Use High Throughput Screening Technology
(HTPS)
• EPA to run ER, AR assays (Tier 1) assays before
priority setting on 15,000 chemicals
• Use data to help set priorities for remainder of
Tier 1 screen
PROPOSED SOLUTION # 2
m Develop and validate QSAR models based on
receptor binding data
-------
Currently investigating 2 approaches
Both address receptor binding
Both need further validation
Will be a data source for EDPSD
Will be continually improved using data
generated by the Screening Program
-------
In vitro Screens
• ER Binding / Reporter Gene Assay
• AR Binding / Reporter Gene Assay
• Steroidogenesis Assay with minced testis
In vivo Screens
• Rodent 3-day Uterotrophic Assay (sc)
• Rodent 20-day Pubertal Female Assay with Thyroid
• Rodent 5-7 day Hershberger Assay
• Frog Metamorphosis Assay
• Fish Reproduction Screening Assay
-------
Rodent 20-day Pubertal Male Assay \N\tir\
Thyroid
Placental aromatase
Rodent in utero through Lactation Assay
-------
Multigeneration reproduction and
development studies
• Mammals
• Avians
• Amphibians
• Fish
• Invertebrates
-------
Overview
Research contri,bd£f§ns.to drotocg
' .- pi ^lA^
Receptor binding/transcriptional adiv
lopmei
screens
* Tissue slice assay (in vitro s™
Hershberger assay (in m
Female pubertaj^ass^p^rats (in vivo screen) \
Male pubertal ass^y in rats (i/f vivo screen)
Developmental toxicity screen in rats (in vivo alternative screen i
Frogju^famorphosis as$ay {in vivo screen)
Jish 21-day reproduction screen in the fathead minnow (in vivo
screen) >£ *
Two generation mammalian reproduction study (in i//V0test)
Invertebrate reproduction assay (in i//V0test) I
Developing position paper/case study
-------
ORD - conducting research, developing protocols,
serving as a consultant on the
standardization/validation
OPPTS - the lead on the standardization/ validation
• Standardization & demonstration of protocol
• Validation in multiple laboratories
• Scientific peer review
EDMVS - advises EPA on standardization and
validation issues
ICCVAM - interagency coordinating committee on
validating alternative methods
-------
\
' V r—
DI.
Method development and preparation of Detailed
Review Paper (DRP)
Pre-validation
• Demonstration of relevance
• Development of standardized protocol
Determination of readiness for validation in
consultation with EDVMS and ICCVAM
Validation in multiple laboratories
Independent peer review and review by ICCVAM
-------
Use Order (FIFRA, FQPA, SDWA) and /or
Test Rule authority (TSCA) to require
industry to develop screening and testing
data
EPA conducts hazard/risk assessment
-------
>crine Disrupters W
ated Under
Insecticide, Fungicide, Rodenticide Act
Toxic Substances Control Act
Clean Water Act
Safe Drinking Water Act
Clean Air Act
-------
-------
-------
ects Research
'si, 2, & 3
Urethral Opening
Evaluates adequacy of current testing
guidelines
Develops new/improved protocols for
screening and testing program
Determines classes of chemicals that
EDCs and their potencies
Determines the dose-response curves
EDCs at environmentally relevant
concentrations
Investigates mode of action of certain
Conducts comparative endocrinology studies
Examines population level effects
Phallus
Prepuce
s
Os penis
Prepuce
Urethral opening
-------
sure Resear
^
Identifies and improves understanding
of major exposure routes and processes
Develops predictive models estimating
the extent and magnitude of exposures
-------
Risk Assessment Research
Developing position paper on how
results from EDSP could be
incorporated into hazard
characterization assessments
Developing case study for methods on
integrating human health and ecological
EDC data into risk assessments
-------
Risk Manaaement
-LTGs1 & 2
M
Identifying major sources of EDCs entering
the environment, with focus o
contaminated sediments
• wastewater treatment plants
• drinking water treatment plants
Developing tools for risk manag
EDCs, such as biodegradation pr
pollution prevention strategies
-------
-------
There is global concern regarding exposures to some
environmental agents that interfere with endocrine
systems
Congressional mandates require that EPA establish a
screening and testing program
EPA is implementing such a program based on
advisory committee recommendations that
incorporates a tiered approach
EPA's research is providing immediate results for
implementing the screening and testing program
-------
EPA's long-term research program on EDCs focuses
on the most critical uncertainties in determining
whether humans and wildlife populations are being
impacted by levels of EDCs in the environment, in
identifying the sources of those exposures, and
approaches to reduce/prevent them
EPA is the only federal agency doing research in the
area of risk management of endocrine disruptors
-------
science for a changing world
Monitoring Endocrine
Disrupting Compounds(EDC's)
in Aquatic Ecosystems
in the U.S.
Steven L. Goodbred
U.S. Geological Survey
California State University
Sacramento, California
-------
No Systematic Monitoring of
just EDC's in the U.S.
• Incomplete consensus on EDC's
• Analytical methods still being
developed for many compounds
• Difficulty selecting sampling matrix
(sediment, water, tissue)
• Major funding needed
-------
Several U.S. Agencies nave
National Monitoring Programs
which include EDC's
• U.S. Geological Survey
(NAWQA, TOXICS, BEST)
• U.S. Environmental Protection Agency
(EMAP, REMAP, National Dioxin Study)
• National Oceanic and Atmospheric
Administration
(Mussel Watch, Benthic Surveillance)
• Food and Drug Administration
(National Monitoring Program for Food and
-------
USGS National Water Quality
Assessment Program(NAWQA)
http://water.gov/NAWQA
Established in 1991
• Assess status of trends of U.S. surface and
ground water
• Elucidate factors that affect water quality
• Rigorous field and lab QA/QC
• Nationally consistent chemical and
ecological data collection and analysis
"USCS
-------
Chemical Monitoring Sites in
USGS's National Water Quality
Assessment Program
uses
-------
Organic Compounds Monitore
in the U.S. by USGS, NAWQA
Water
(pesticides)
Bed Sediment
(PAHs, phenols, etc.)
Fish Tissue
(OCs, PCBs)
Total
Suspected EDO's
* from Keith, 1997
#EDC'
USGS
-------
Pesticides widespread in
streams & ground water
Streams
Aaricultural Areas
Water
Shallow Ground Wate
Streams
Jrban Areas
Shallow Ground Water
lixed Land Use
Major Rivers • -
and Streams water
Major Aquifers
85%
92%
59%
1
99%
49%
96%
100%
33%
% Samples with One or More Pesticides
-------
Pesticides almost always
occur as mixtures
100
Urban
Agriculture
Mixed Land Use
Streams
Ground Water
Number of Compounds
-------
Pesticides in streams usually
occur in strong seasonal pulses
-------
Breakdown products often total
10-25 times the concentration of
parent compounds
Herbicide breakdown products
Herbicide parent compounds
-------
Reconnaissance data show potential of
pesticides to affect fish endocrine systems
Carp in 11 U.S.
Streams
Females
Total pesticides (ug/L)
-------
Herbicides in agricultural
streams were highest in the
Corn Belt
CONCENTRATIONS \
Highest
-------
Herbicides that are EDCs
in U.S. Surface Water
LandD
use
#ofa
samples
DetectionD Percentile (ug/L)D
frequency 50% 95%
ATRAZINE
Maxim umD
(ug/L)
1559D
85%D
3.51D
120D
Urban
SIMAZINE
UrbanD
USGS
-------
Organochlorines and PCBs
in Fish from the U.S.
LandD # ofD
use samples
UrbanD
Forest
UrbanD
Forest
171D
171D
DetectionD Percentile (ug/kg WW)D MaximumD
frequency 50% 95% (ug/kg WW)
P,P' DDE
38%
43D 3,080D
I 160D
-------
PAHs in Bed Sediment
from the U.S.
LandD # ofD
use samples
UrbanD
Forest
131D
|D 220D
UrbanD 114D
Forest 11 f
DetectionD Percentile (ug/kg WW)D
frequency 50% 95%
PYRENE
89%D
21 %D
26%
41OD 4,840D
-------
What can we conclude
about EDC's from NAWQA
• Several suspected EDC's widespread in U.S.
streams, surface water, sediment and fish
• Occurrence follows land and chemical use
• Many EDC's at low levels(
-------
science for a changing world
A National Reconnaissance of
Pharmaceuticals and other
Emerging Contaminants in U.S
Streams
Toxic Substances Hydrology Program
-------
-------
Project Goal
Determine if Pharmaceuticals,
antibiotics, hormones, and other
wastewater related compounds ar
entering the environment through
human, animal, and industrial
waste waters.
uses
-------
pproac
• Develop sensitive and specific
analytical methods
• Evaluate environmental occurrence
in "susceptible" waters
USGS
-------
Target Chemicals (95 OWCs)
1 22 Antibiotics (lincomycin, trimethoprim)
1 14 Prescription Drugs (fluoxetine,
gemfibrozil, ranitidine)
• 5 Nonprescription Drugs (caffeine)
• 15 Hormones and Sterols (equilin)
39 Household and Industrial
Compounds (triclosan, bisphenol A)
USGS
-------
Target Hormones and Sterols
ioaenic Hormones
17b-Estradiol
17a-Estradiol
Estrone
Estriol
Testosterone
Progesterone
cis-Androsterone
Equilenin *
Equilin *
Synthetic Hormones
17a-Ethynylestradiol
Mestranol
19-Norethisterone
Sterols
Cholesterol
3b-Coprostanol
Stigmastanol
(plant sterol)
* Hormone replacement therapy; commonly prescribed
** Ovulation inhibitor
-------
Potential Sources
WWTF
On Site Septic Systems
Animal Feeding Op.'s
Industrial Discharges
(Medical)
USGS
-------
Occurrence in Streams Susceptible
to Sources of OWCs
139 streams in 30 states
• 62 Agricultural
• 52 Urban
• 17 Mixed Land Use
• 8 Minimally developed
USGS
-------
Stream Monitoring Network (1999-2000)
N jr
Urban
Agricultural
Mixed
Minimally developed
-------
^mi
-------
General Results
OWCs found in most streams sampled (-80%)
82 of 95 OWCs detected, representing a wide
range of uses and origins
Measured OWCs concentrations generally low
- few standards or guidelines exceeded (few exist)
Detection of multiple OWCs common
- about 75% had more than 1 OWC
- about 35% had more than 10 OWCs
USGS
-------
Number of OWCs Detected
520
(131)
Highly
Susceptible
[80.9%]
Less
Susceptible
[62.5%]
uses
-------
Aggregate Concentrations
(131)
c 30
6 20
Highly
Susceptible
[80.9%]
Less
Susceptible
[62.5%]
uses
-------
(82%) (69%) (64%) (48%) (32%)
(89%) (74%) (66%) (60%) (41%)
Detection
Frequency
Concentration
USGS
-------
Removal throuah WW Treatment ?
i Influent
i Biological filter!
Stumpf, et al, 1999
-------
Is Endocrine Disruption Widespread?
DOI Study Sites Showing Evidence of
Endocrine Disruption in Wildlife
JV „,-/
n?
—<
O
O
J»t— .1
N 9?
N "I
O Biomarker effects
D Biological endpoint effects
X" Population level effects
•^Q
Gross etal. 2000
-------
For More Info, Try the Internet
The National Reconnaissance:
toxics.usgs.gov/regional/emc.html
The Toxics Program:
toxics.usgs.gov
The USGS:
www.usgs.gov
USGS
-------
ENDOCRINE DISRUPTORS FROM COMBUSTION AND
VEHICULAR EMISSIONS: IDENTIFICATION AND
SOURCE NOMINATION
Brian Gullett1*, Jeff Ryan1, Paul Lemieux1, Carolyn Acheson2,
Michael DeVito3, James Rabinowitz3, Sukh Sidhu4,
Richard Striebich4, Joy Klosterman4
U.S. Environmental Protection Agency
National Risk Management Research Laboratory, Air Pollution Prevention and Control
Division (MD-65), Research Triangle Park, NC 27711
2National Risk Management Research Laboratory, Cincinnati, OH 45268
3National Health and Environmental Effects Research Laboratory, Toxicology Division,
Research Triangle Park, NC 27711
4University of Dayton Research Institute, 300 College Park, Dayton, OH 45469
*919-541-1534, 919-541-0554 (fax), gullett.brian@epa.gov
-------
BACKGROUND
A significant fraction of organic
emissions from combustion sources
remains uncharacterized
100's of compounds are emitted
- e.g., PAHs, oxygenates, alkyl
phenols
We only see what we specifically
look for
- e.g., PCDD/Fs "dioxins"
EDCs are emitted from combustion
sources
- e.g., PCDD/F, PCB, Hg
Combustion sources are ubiquitous
- constitute a major exposure source
-------
GAS PHASE, AROMATIC PRECURSORS -
How do we get complex aromatic structures?
Benzene
CH(S)+CH
22
C2H4+C2H3
Simple,
low-C-
number
HC's
+H2CCCH
H,CCCH+CH*-^
&, «5
CH2CHCHCH2
"H- '+H
H2CCCH3
3 5
+H
(allene)
+H
allyl)
i
-H
(C=C-C=C and C-C=C=C)
v-H +
H2C=C=CR H2C-C=CR
R=H (propargyl)
R=CH,(1-methylallenyl)
+H2CCCH or
+H2CCCH3
Branched Aromatics
After Marinov, Pitz, Westbrook, Castaldi and Senkan, 1996
-------
OBJECTIVE
We will survey combustion sources for potential endocrine
disruption activity, try to isolate the compounds
responsible for such activity, and attempt to estimate their
emission factors.
-------
APPROACH
Opportunistic combustion source sampling
- domestic waste burning, diesel trucks (HDDVs), forest fires,
fireplaces, and woodstove....others.
Bioassays
- Yeast estrogen assay
- CALUX
- Vitellogenin mRNA Assay
Sample fractionation to isolate target compounds
- HPLC technique (L. Brooks)
- Capillary Electrophoresis
- TIE method (G. Ankley)
Multi Dimensional Gas Chromatography
- diagram/description
QSAR, Statistical analyses
- Structure clues
-------
Source Characterization
Done • Future
- diesel truck - pine straw
- woodstove - structural fires
- sewage sludge - oil spill fire
incinerator - municipal waste, full
- domestic waste scale
- DoD diesel truck
-------
On-Road Diesel Sampling
Dioxin Sampling
Train
Heated
Filter
Box '
Glass
Tube
High Volume
Vacuum
Pumps
Plastic Nut
with Teflon
Washer
Orifice
Dry Gas Meter
^) Thermocouple
(Q Differential
Pressure
Transmitter
(^ Vacuum Gauge
Pump 1/2 lnch
H Teflon
Tubing
Pump
Filter
/ Ice
Cooler Bath
SS
Water
_ice Jacket
T Water
XAD-2
Module
Water
I Knockout
Coiled
SS
Tubing
-------
"Uncontrolled" Burning Sources
-------
Burning of
Domestic Waste
-------
Wheat Straw Testing
-------
CA PCDD/F, PCB
Sources?
FREPLACE
WOODSTOVE
-------
SAMPLE CHARACTERIZATION
Yeast Estrogen Screen
IVIed i LJ m
-------
Environmental Estrogens Responding to
Yeast Screen
Row A = Bisphenol
Row C = Genistein
Row E = Nonyphenol
Row F = Octylphenol
Row H = 17(3-Estradiol
Rows B,D,and G (blank)
Deep Red Color indicates
Estrogenic Activity
Yellow indicates
Background ((3-gal)
IMMtMttlt
m
V
ui
mt • • •
-------
Gas Chromatography/Mass Spectroscopy
a
«
ffS
342-
3.00% n
416-
5.00% 1
30:00
Estrone
17p-Estradiol
17a-Ethynylestradiol
31:40
33:20
3SKW
36:40
38:20
Time (min)
-------
Combustion Pollutant Analysis via Multidimensional Gas
Chromatography (courtesy of UDRI)
Conventional
GC-MS
MDGC heartcrft
of region
Cryotrap and
release
30m
Non-polar
Primary
column
I
4 m
Polar
secondary
column
MSB
39.80 39.90 40.00 40.10 40.20 40.30 40.40 40.50 40.60 40.70 40.80 40.90 41.00
Multuinensio
-------
RESULTS
Examples of oxygenates in diesel extracts using MDGC-MS (scanning
mode)
*o
Benzaldehyde
HC-
sO
Methylbenzaldehyde
'Hydroxybenzaldehyde
•Naphthalene dione
-OH
-CH-,
O
O
Dibenzofuran
'Hydroxybiphenyl
Xanthene
OH
•Naphthalene-
carboxaldehyde
SO
-------
RESULTS
Examples of Oxygenates from Barrel Burn Extract
•hydroxy
cyclopentanone (7)
•benzaldehyde (87)
•substituted
cyclohexanone (72)
•phenol (80)
•methyl quinol (90)
•alkyl phenols
(72 - 91)
•methoxy methyl
phenol (93)
•hydroxy phenyl
butadiene (64)
( ) denotes Match Quality out of 100
•benzene ethanol 3-
methoxy (52)
•benzoic acid (64)
•4-ethyl-2-methoxy
phenol(90)
•benzendiol (90)
•dimethyl anisole (27)
N
•eugenol (96)
•ethyl vanillin (5)
•methoxy propenyl
phenol(90)
•dihydroxy dimethyl
benzaldehyde (5)
•long chain carboxylic
acids
•Hydroxy methoxy
phenyl ethanone (72)
•hydroxy methoxy
phenyl propanone
•methyl
•methoxy propyl
phenol (64)
•vanillin (78)
-------
RESULTS
Examples of PAH from Barrel Burn Extract
•fluoro-biphenyl (72)
•biphenyl (83)
•me-biphenyl (80)
•dimethyl
phenanthrene (64)
•tetramethyl
phenanthrene (38)
( ) denotes Match Quality out of 100
•biphenylene (72)
•acenaphthalene (80)
•bibenzyl (74)
•phenanthrene or
anthracene (72)
•fluoranthene or
pyrene (72)
•vinyl naphthalene
(48)
•fluorene (90)
•methyl fluorene
(74)
•phenyl naphthalene
(69)
•methyl
phenanthrene (43)
-------
RESULTS
Examples of Oxy-PAH from Barrel Burn Extract
( ) denotes Match Quality out of 100
•benzofuran (83)
•benzophenone (87)
•methyl benzofuran (72)
•fluoren-one (72)
•phthalic anhydride (86)
-------
ENDOCRINE DISRUPTORS FROM COMBUSTION
AND VEHICULAR EMISSIONS: IDENTIFICATION AND
SOURCE NOMINATION
Brian Gullett1*, Clyde Owens1, Jeff Ryan1, Paul Lemieux1,
Carolyn Acheson2,
Michael DeVito3, James Rabinowitz3,
Sukh Sidhu4,
Richard Striebich4, Joy Klosterman4
»wi#
U.S. Environmental Protection Agency
National Risk Management Research Laboratory, Air Pollution Prevention and Control
Division (MD-65), Research Triangle Park, NC 27711
2National Risk Management Research Laboratory, Cincinnati, OH 45268
3National Health and Environmental Effects Research Laboratory, Toxicology Division,
Research Triangle Park, NC 27711
4University of Dayton Research Institute, 300 College Park, Dayton, OH 45469
*919-541-1534, 919-541-0554 (fax), gullett.brian@epa.gov
-------
Fate of the Endogenous Hormones
17p-Estradiol and Testosterone in
Composted Poultry Manure and their
Sorption/Mobility in Loam Soil and Sand
Heldur Hakk a, Patricia Millner b, and Gerald Larsen a
Colleen Pfaff a, Barb Magelky a, and Frank Casey c
a USDA-ARS Biosciences Research Lab, Fargo, ND
b USDA-ARS Soil Microbial Systems Lab, Beltsville, MD
c North Dakota State University, Dept. Soil Science, Fargo, ND
o
-------
Estradiol and Testosterone
-------
Estradiol
Native hormone for estrogen receptor
Most potent steroidal estrogen hormone
104-106 times as potent as active APP's
Excreted in mammals/birds in urine/feces
Derivatives administered as growth
promoters
Carcinogenic (breast cancer)
-------
Toxicology of Estradiol
5-500 ng/L E in water increased alfalfa growth
5000-500,000 ng/L E decreased alfalfa growth
300 ppb E in poultry litter fed to heifers caused
premature udder development
1-10 ng/L E increases vitellogenin, inhibits
testicular growth in trout, feminization
50-300 ppb E resulted in halted gonads and
feeding, and death in brown trout
250-5000 ng/L E were 84-100% female salmon;
10-200 ug/L contributed to death
-------
Testosterone
Native hormone for androgen receptor
Excreted in mammalian/avian urine and feces
Responsible for developing and maintaining
male sex characteristics
Reduced to 5a-dihydrotestosterone, which
binds receptor with higher affinity
Serves as a prohormone [T to 17(3 -E2 (brain)
and T to 5|3 -diOH T (kidney)]
Androgenicity of native waters leads to
masculinization of mosquitofish, killifish (FL)
-------
Excretion Levels
Adult cattle excrete 30 mg E/day (13 ng/L)
- Growth hormones increase 5-6 fold in
urine
Chickens excrete 44 ng/g (Ave. dry wt.)
- 6000 g/yr from chickens on E. shore MD
Women excrete 25-100 ug/day at ovulation
- 30 mg/ day El9 E2, and E3 excreted at
pregnancy
- Estrogen replacement therapy prolongs
Men excrete 2-25 ug/day
-------
Selected Environmental Levels of E and T
£-\J\J\J
2000 -
1500 -
1000/
400^
300 -
200 -
100 -
n
Field Runoff
I
Effects on fish (E)
Israeli Sewage
i
• m=
i
Pasture
i
us
I I Estradiol
i i Testosterone
Sewage
NWA Aquifers
I I
Sea of Galilee AR Municipal
-------
The Process of Aerobic Compostin
Microorganisms consume O2 while feeding
on organic matter
Generates heat, CO2, and H2O vapor
Feedstock can be manure, sludge, leaves,
paper, food waste
Carbon is stabilized
Finished compost is soil-like, odorless, easy
to handle, good storage properties
-------
Organic Matter
(including carbon,
chemical energy,
protein, nitrogen)
Minerals/ Nutrients
Water
(40-65%)
Microorganisms
(bacteria, fungi,
actinomycetes)
Heat
Water
HOI
jost Pil
Decomposed/
Stablized
Organic matter
-------
ARS-BARC Composting Center
2 acres
8" thick pad
- Coal ash, cement kiln dust, quicklime, cement
Surrounded by 8 acres orchardgrass buffer
Grass and tree buffer zones
Chesapeake Bay watershed
-------
Composting Center, Beltsville, MD
•r .,
-------
Composting Methods
Chicken layer manure obtained from
commercial producer in MD
Manure adjusted to 30:1 C:N
- Manure (3.3 parts), old hay (2), straw (2)
partially decomposed leaves (4), starter
compost (2); [Christiana clay (1)]
Moisture content maintained at 60%
Windrows were 160' x 5' x 40 inches high
Windrows turned weekly to aerate
Three subsamples removed from center
weekly (3 wks) then biweekly (16 wks)
-------
Windrow Turner
-
'
• '•
••
-
-------
Frozen at -20
Sample
Preparation
ELISA
r- >
Homogenize
with dry ice
200mg compost/
50ml water
Centrifuged 15 min
@ 2000rpm
-------
Compost pile parameters
nco2
Temp
-------
Testosterone Degradation
1.2
.0
'*J
1 i
0.8
0.6 +
o o
^ 0.4
I 0.2
0
0
k= -0.0752/day
20
4Q
60
80
Time (d)
100
120
140
-------
Estradiol degradation
c
_g
ra
^^m
4-1
c
£ 0
o /*
c i£
o 0
O
CD
ca
0)
1.4 -i
1.2-
1 ^
0.8-
0.6
0.4
0.2
0
k= -0.0121/day
. T
^r \ ~~~^^
I ^^\ ^ s
""""\\. 5_
"^~~~~^_^
-^^^^
^^ ^~-^___^
^ — — ^
1 II III 1
0 20 40 60 80 100 120 140 160
Time (d)
-------
JO
CD
1.2
1 +
0.8
0.6 -\-
0.4
0.2
0
0
20
Testosterone degradation w/o Clay
k= -0.0752/day
40
60 80 100 120 140
Time (d)
Relative Concentratic
C/Co
1 9
1 «
0.8
0.6 .
0.4
0.2
Testosterone degradation w/o Clay
\ k= -0.0653/day
0 20 40 60
80 100 120 140 160
Time (d)
-------
O 80
£ 60
3
2 40
20
0
Compost Pile Temperatures
(24" depth)
Thermophilic Mesophilic
I
6 10 13 21 41 45 55 70 73 74 75 116
Day
-------
Commercial By-Product
Derived from Solid Wastes
Cattle
Swine
Municipal
Estradiol (ppb)
19±3.5
152 ±2.4
31 ±5.4
Testosterone (ppb)
n.d.
21 ±18
29 ± 7.3
-------
Conclusions
Removable levels of composted chicken
manure hormones, estradiol and
testosterone, into water were initially 100 &
200 ppb (d.w.), respectively
Estradiol degradation was observed in
composted chicken manure with time
Testosterone was more rapidly degraded
(5-6 fold) than estradiol
-------
Conclusions (Cont'd)
Two rates of degradation may exist,
rapid during thermophilic temperatures,
slower during mesophilic
Neither hormone was degraded
completely after 19 weeks, when curing
was completed
Christiana clay amendment did not affect
rate of degradation of either hormone
-------
Conclusions (Cont'd)
Composting may provide an
environmentally friendly method of
reducing, but not eliminating, the
introduction of potent, endogenous
hormones into the environment
-------
Batch Studies
1.6gsoil/8mlof0.01CaCL
at various concentrations;
Spike with 14C-hormones
Continuous rotation
for 7 days; aliquots
taken at 2, 4 and 7 day
-------
Glass column
packed with
Silt Loam Soil
or Sand
8.5cm diax 15cm
Soil: 1126gms
Pore Volume 455ml
Sand: 1427gms
Pore Volume 303ml
-------
[14C]Estradiol and Testosterone at various soil column depths
i u -
-
c
E
-------
400000
300000
200000
100000
Estradiol Sand Column
500 1000 1500 2000 2500 3000 3500
Cumulative Volume (ml)
-------
Preliminary Metabolism Data (tic)
Soil
Sand
Estradiol
2 metabolites
(17-26%)
1 metabolite (85%)
74-83% bound
Testosterone
1 metabolite
(11-32%)
Parent (88%)
Parent (5-17%)
50% bound
-------
Batch Studies
Column Studies
D)
_§
I 3
ro
T3
0
.Q
2 -
1
Testosterone
1.0
0.8 -
ID)
D)
_§
.1 0.6 H
0
o
c
T3
0
0.4 -
o « «
w 0.2 -
0.0
0.00
Experimental values
Freundlich isotherm
8 10 12 14
-1,
16
Dissolved concentration (mg L )
Estradiol
Experimental values
Freundlich isotherm
0.05 0.10 0.15
Dissolved concentration (mg L" )
0.20
O
Q.
CD
Q
10 -
0 14C labeled estradiol
• 14C labeled testosterone
Model fit
5 C 32C d C apfe
dt
~~ V
dx 0
[KdC-s\
0.0 0.1 0.2 0.3 0.4 Q.I
Relative Concentration (C/Co)
-------
Kd Values
Estradiol
Testosterone
Columna
1661
182
Batchb
4388
348
a Column flow approximately one dimensional
b Batch soil dispersed by agitation -
more soil/chemical interaction
-------
Conclusions
Estradiol and testosterone are readily
transported though sand (i.e. not sorbed to
the soil mineral fraction)
Estradiol and testosterone are rapidly and
strongly sorbed to the soil (i.e. sorbed to the
soil organic fraction)
Preliminary data indicate estradiol and
testosterone may be metabolized in soil
Kd values derived from batch studies were
comparable with soil column results
-------
Future Work
Express degradation data on a combustible
carbon basis to normalize
Conduct degradation studies with laboratory
composters using radiolabeled hormones to
determine chemical fate of hormones
Prolong thermophilic heating phase in order
to increase overall degradation
Conduct composting studies with swine and
cattle manure
-------
^^M
Potential of Concentrated Animal Feed Operations (CAFOs)
To Contribute Estrogens to the Environment
Principal Investigator
Stephen R. Hutchins
USEPA NRMRL, Subsurface Protection and Remediation Division
.Bohgrt SJepa.gov
-------
Concentrated Animal Feed Operations (CAFOs)
• In the United States, an estimated 376,000 animal feed operations
confine animals, generating approximately 128 billion pounds of manure
each year
• A facility is an animal feed operation (AFO) if animals are
stabled/confined, or fed/maintained, for 45 days or more within any
12-month period, and the facility does not produce any crops, ™~
vegetation, or forage growth
• Concentrated animal feed operations (CAFOs) are the largest of these
and are regulated under the Clean Water Act. CAFOs are generally
considered to be operations with more than 1000 animal units (AU)
^^M
Manure Available for Land Application, 1997
Sector
Cattle
Dairy
Swine
Poultry
Total Manure
(billion pounds)
32.9
45.5
16.3
33.5
Percentage Share
by > 1000 AU Operations
83%
23%
55%
- -^-r/K- 49%
'Office of Wastewater Management (Office of Water/USEPA) Website:
(http://cfpub1.epa.gov/npdes/home/cfm)
-------
Environmental Estrogens
Refer to a wide range of anthropogenic or naturally occurring
compounds that elicit estrogenic responses by mimicking
endogenous estradiol
Natural Estrogens:
- 17p-Estradiol (Estradiol ), Estnol, Estrone, Equilm, Equilenm, Genistem
Synthetic Estrogens:
Ethinyl estradiol, Mestranol, Diethylstilbestrol
ther Compounds
o, p'- DDT, Nonylphenol, Bisphenol A
-------
p
-------
Comparison of Estrogenic Activity in Terms
of EC50 Measured by Yeast Estrogen Screen
Substance
Natural Estroaens
Estradiol
Estrone
Estriol
17p-Estradiol-3-Sulfate
Ph ytoestrogens
Genistein
Other Comoounds
Nonylphenol
Bisphenol A
"Matsui et al, 2000
Relative Ratio of Estrogenic Activity
0.21
0.0013
0.000053
0.00011
0.001
0.00027
-------
Comparison of Estrogenic Activity in Terms
of Plasma Vitellogenin Induction*
Substance
Natural Estroaens
Estrone
Estradiol
Minimum Aqueous Concentration
Required for Vitellogenin Induction
100ng/L
10ng/L
Synthetic Estroaens
Ethinyl Estradiol
rArcand-Hoy et al, 1998
2ng/L
-------
Use and Expected Environmental Impact
of Estrogenic Pharmaceuticals
Prescribed and Sold in the U.S.*
Pharmaceutical
Estrogen
Product
Human Use
Calculated
Estrogen
Use
Expected Introduction
Concentrations to the
Aquatic Environment
Oral Contraceptives
(ethinyl estradiol, mestranol)
Hormone Replacement Therapy
(conjugated estrogens)
88 kg/yr
1700 kg/yr
2.2 ng/L
42 ng/L
Animal Use (Cattle Onl\
Growth-Enhancement
(estradiol)
580 kg/yr
14 ng/L
Arcand-Hoy et al, 1998
-------
CAFO Contributions of Estrogens - Cattle
Estimated that at least 90% of feedlot cattle slaughtered in 1995
were administered growth-enhancing hormones
Growth Hormones
- Androgens (trenbolone acetate, testosterone propionate,
-—-— - ' .*#***•
- Progestins (progesterone)
• For cattle, the estradiol concentration in the urine averages 13 ng/L
Cattle subjected to growth hormones generate urine with estradiol
concentrations five- to sixfold greater
m
m
-------
CAFO Contributions of Estrogens - Poultry
• No growth hormones added: natural production of estrogens and
testosterone
* In 1998, the U.S. poultry industry produced almost eight billion
b.rqijers with a total production of almost 12 billion kg litter
Average estimated hormone concentrations
14//g estrogens (estradiol, estrone) in male broilers
weight litter:
65//g estrogens (estradiol, estrone) in female broilers
133 //g testosterone in male and female broile
Estimated estrogen production: 160,000 - 760,000 kg/year
ield study shows sizeable edge-of-field losses of estradiol
!0-2530 ng/L) and testosterone (10-1830 ng/L) in runoff from
litter-amended grasslands (Finlay-Moore et al, 2000)
-------
CAFO Contributions of Estrogens - Swine
-------
^^M
-------
Evaluation of CAFO Lagoon Effluents
for EDC Activity using Bioassays
Principal Investigator
James N. Dumont
Department of Zoology
430 Life Sciences West
Oklahoma State University
Stillwater, OK 74078
Phone: (405)744-9683
Fax: (405)744-7824
email: dumontj@okstate.edu
Co-Principal Investigator
David M. Janz
Department of Zoology
430 Life Sciences West
Oklahoma State University
Stillwater, OK 74078
Phone: (405)744-7593
Fax: (405)744-7824
email: djanz@okstate.edu
-------
OSU CAFO EDC Study - Objective
The objective of this study is to evaluate lagoon samples from swine,
beef, and dairy CAFOs for possible EDC activity, using a variety
of tests based upon the African Clawed Frog (Xenopus laevis)
OSU Swine Lagoon
OSU Beef Lagoon
OSU Dairy Lagoon
-------
OSU CAFO EDC Study - Test Description
FETAX
(Frog Embryo Teratogenesis Assay -Xenopus)
\
XTRA
(Xenopus Tail Resorption Assay)
Adult Male Frog Exposure -
Vitellogenin Expression
and Estradiol/Testosterone Changes
i
RJ*
-------
OSU CAFO EDC Study - Test Description
FETAX - Frog Embryo Teratogenesis Assay - Xenopus
Standardized Protocol (ASTM Standard G
9-91)
96-hr assay that uses early stage Xenopus embryos (Stages 8-1
approximately 8-12 hours old)
Measures growth, mortality, and malformation for detection of
potential developmentally toxic compounds and mixtures
Not used for detection of EDC activity per se; needed for
screening samples for potential toxicity to determine allowable
concentrations
-------
AFO EDC Study - Test Description
TRA - Xenopus Tail Resorption Assay
Tail resorption controlled by thyroxin
thyroid gland
- measures disruption of
Uses Stage 56/57 (approximately 38/40-day old) Xenopus larvae
Four replicate tanks, 10 larvae per tank, water changed twice per week
JBr
Larvae photographed every other day; tail length measured by
SigmaScan software
-------
OSU CAFO EDC Study - Test Description
Adult Male Frog Exposure - Vitellogenin Expression and Estradiol/Testosterone
Changes
• Vitellogenin Analysis - Exposure of male oviparous vertebrates to natural
and synthetic estrogens can induce synthesis of the
phospholipoglycoprotein yolk precursor Vitellogenin
• Estradiol/Testosterone Analyses - Indicator of alterations in reproductive
endocrine homeostasis
Exposure: Four groups of 5 adult male Xenopus exposed for 21 days
• Untreated Controls - reared in charcoal-filtered water
• Positive Plasma Controls - reared in charcoal-filtered water;
intraperitoneal injection of 1 mg/kg ethinylestradiol on Days 1, 3, and 6
• Positive Aqueous Controls - reared in charcoal-filtered water with 1 mg/L
ethinylestradiol
• Test Group - reared in CAFO lagoon effluen
Analyses
• After exposure, plasma prepared and assayed for Vitellogenin using Western
immunoblotting and enzyme-linked immunoassay (ELISA), and assayed for
estradiol and testosterone using enzyme-linked immunoassay (ELISA)
-------
uay - conclusions
ions are Preliminary - Analyses Pei
Although the swine effluent lagoon i
exhibited significant EDC activity, at least based on these bioassays
1
However:
These lag
operations
mam
EDC effects on steroid hormone homeostaesis may be more pronounced
under long-term exposure
-------
OSU CAFO EDC Study - Poster Presentation
22nd Annual
C Meeting, November 11-15, Baltimore, MD
Lagoon Water from Confined Animal Feed Operations and
Amphibian Development
Dumont, J.N.*, Oklahoma State University, Stillwater, OK
Hutchins, S.R., U.S. EPA (NRMRL/SPRD), Ada, OK
..-
Endocrine Modulating Effects of Lagoon Water from Confined
Animal Feed Operations on Amphibians
Weber, L.P.*, Dumont, J.N., and Janz, D.M., OSU, Stillwater, OK
Selcer, K.W., Duquesne University, Pittsburgh, PA
Hutchins, S.R., U.S. EPA (NRMRL/SPRD), Ada, OK
-------
-------
Analysis of Environmental Estrogens in Swine
Wastewater, using ELISA, LC/MS/MS, and GC/MS
G. Peter Breidenbach
Research Microbiologist
ManTech Environmental Services, Inc.
Robert S. Kerr Environmental Research Center
P.O. Box1198
Ada, OK 74820
Phone: (580)436-8668
Fax: (580)436-8501
email: breidenbach.peter@epa.gov
Dennis D. Fine
Analytical Chemist
ManTech Environmental Services, Inc.
Robert S. Kerr Environmental Research Center
P.O. Box1198
Ada, OK 74820
Phone: (580)436-8669
Fax: (580)436-8501
email: fine.dennis@epa.gov
-------
-------
•
-------
TOOO iflt
PYRfcX*
-------
Environmental Estrogen Analysis - Objectiv
• The objective of this study is to develop a protocol for screening
and analyzing swine lagoon effluent and ground water for
estrogens at environmental levels (ng/L)
creening will be done using enzyme-linked immunoass
(ELISA) specific for estradiol. Positive samples will then be
analyzed for individual estrogens
Individual estrogens will be analyzed by LC/MS/MS
LC/MS/MS Interferences - switch to GC/MS
-------
Environmental Estrogen Analysis - ELISA Screen
Initial Procedu
Solid phase extraction of 250-mL sample using 6-1
VI-CARB SPE
1i^^m* i—«-. ,u
equentiai
ng with
Elution with methylene chloride/methano
Evaporation to dryness; resuspension with water/methanol to 500 juL
Direct analysis of 20-jL/L aliquots by ELISA for estradi
Estimate of estradiol concentration using external calibration cr
-------
Environmental Estrogen Analysis - ELISA Screen
,marry Findings
Cross-reactivity: higher concentrations of other estrogens wii
produce a similar positive response in the estradiol ELISA scree,
"'*""* \
- Estriol - Response is 1% that of estradiol
Estrone - Response is 1% that of estradiol
Ethinyl estradiol - Response is 0.2% that of estradiol
appears to exert a positive intei i
in that estimated estradiol concentrations can be much higher than
those confirmed by direct LC/MS/MS analysis
-------
Environmental Estrogen Analysis - LC/MS/MS Analysis
Initial Procedure (Finnegan TSQ 7000 Mass spectrometer)
same SPE extr
for ELISA
enmg
. sample loop injection onto 5-pL Zorbax C18SB pack*
column
• ~5l^
Gradient elution with acetonitrile/water
Addition of ammonium hydroxide to column eluent prior to
electrospray source to abstract phenoxy proton from the estrogen
Quantitate estrogen concentrations using calibration curves wit
estrone-d, as the internal standard
-------
Preliminary Results of Swine Lagoon Estrogen Analysis
(all concentrations in ng/L original water sample)
Sample
Type
Lagoon
Well
Lagoon
Well
Lagoon
Well
Lagoon
Lagoon
Well
Lagoon
Lagoon
Lagoon
Well
Lagoon
Lagoon
ELISA
estradiol
27.2
0.1
69.0
1.3
21.4
0.3
26.6
31.2
81.4
22.8
0.3
84.8
56.0
LC/MS/MS Analysis
estriol estradiol ethinyl estrone
estradiol
Not Analyzed
-------
Environmental Estrogen Analysis - LC/MS/MS Analysis
Potential Problems
LC/MS/MS analysis shows numerous organic compounds that elute in
early part of chromatograph that may cause positive response with
ELISA screen - i^^y,^ .
Estrogenic response in LC/MS/MS (electrospray) system is suppressed
when high organic interferences coelute with compounds of interest
(insufficient clean-up)
Possible Solutions
LC clean-up of complex samples (e.g., swine lagoon effluents) prior to
ELISA screen, using gel permeation chromatography and silica gel
prep to remove early-eluting interferences
Evaluate other SPE cartridges and/or alternate analytical techniques
-------
So...What did you
finally end up with??
-------
Environmental Estrogen Analysis - SPE Method
Final Procedur
Solid phase extraction of 500-mL water sample or 25-mL swine lagoon
-"' - -A --mple using OASIS HLB SPE cartridge
equentiai
ammonium hydroxide
Elution with MTBE/methanol
for ELISA screen, resuspend with methanol to 250 jt/L and dilute 1:2
with water
ior GC/MS analysis, resuspend with acetone to 1000 uL
-------
Environmental Estrogen Analysis - ELISA Screen
ocedure - used w
les on
analysis of 20-//L aliquots by ELISA for estradiol
Estimate of estradiol concentration using external calibration curve
O
Concentration factor = 1000; detection limit ~ 0.05 ng/L estradiol in
original water sample
-------
Environmental Estrogen Analysis - GC/MS Analyst
Final Procedure (Finnigan 4600 Mass Spectrometer)
Ht
• Prepare pentafluorobenzyl derivatives of phenolic groups and
trimethylsilyl derivatives of hydroxy g
Analyze the derivatized estrogens by GC/MS using a J&W DBS-MS
capillary column and negative ion chemical ionization mass
spectrometry
Quantitate estrogen concentrations using internal calibration curves
and estrone-d4, estradiol-d3 and ethinyl estradiol-d4 as internal
standards
oncentration factor = 25 (lagoon effluent) and 1000 (ground water);
detection limit ~20 ng/L estrogen in lagoon effluent and ~0.5 ng/L in
ground water
-------
Recovery of Estrogens Spiked in Distilled Water
and in Swine Waste Lagoon Samples
% Recovery
Distilled Water Distilled Water
Spiked at 2 ng/L Spiked at 1 ug/L
7-a-Methylestrone
(surrogate)
110
85.2
Lagoon Effluent
Spiked at 1 ug/L
(duplicates)
86.2 87.4
Estrone
110
83.2
68.8 58.8
Estradiol
160
84.0
73.6 83.2
Ethinyl estradiol
86.0 88.0
Estriol
210
115
75.4 109
-------
Environmental Estrogen Analysis - GC/MS Analysis
tinuing
improve quantitation
ivatizing reagents to
Improve recoveries of estrogens by using deactivated glassware and
increasing concentrations of derivatizing agents
-------
-------
-------
i
ogical Indicators of Exposure of
vatic Organisms to EDC's
Greg Toth
NERL-EERD
January 29, 2002
-------
. m
home messages
* We've developed a specific molecular indicator
of exposure to estrogens in the laboratory
* We've measured this indicator in field studies
* We're developing multi-stressor indicator
methods using DNA microarray technolo
-------
tions
* What is the extent of exposure of wildlife to EDCs?
- Mixtures
4- Surface waters and sediment
- Fish and invertebrates
- Local and regional scale assessments
* What is the linkage between exposure indicators and
effects?
* How effective are risk management practices in
reducing EDC s?
-------
caches
* Use of molecular biology to develop new,
highly-selective, highly-sensitive indicators
Measure changes within cells of organisms exposed
to EDO's
* Measure changes in expression of induced genes
* Collaborate with scientists in the NRMRL,
egio
i
interpretation of indicators in field studies
-------
•urces
* Core ORD facility for molecular biology
- 9 Federal scientists work on gene expression
+ 4 Ph.D.'s I
2 Molecular biology
- I Aquatic biology
- I Biochemistry
+ 2 M.S.'s
Molecular biology
- Advanced equipment / laboratories / contractors
4- 3 Sequencers
4 DNA microarray scanner
-------
Expression Basics
DNA
molecule
Nucleus
Cytoplasm
Gene
activated
• Messenger
RNAs
Specific
proteins
Change in cell behavior
-------
Study Themes
* Single stressor / single
known gene
* Multiple stressors /
multiple genes
IDMA
Ino ecu e
Nucleus
Cytoplasm
Gene
activated
Messenger
RNAs
Nucleus
Cytoplasm
Gene
activated
- Messenger
"RNAs
-------
"enf Studies
* Single Gene Indicator Studies
Laboratory - Adult male and larvae
fathead minnows
Vitellogenin
li]
-------
^gen-induced Expression of
"itellogenin
t' I^JlBTITii
no ecu e
Estrogen
receptor
molecule
Nucleus
Cytoplasm
4-Gene
activated
/ Messenger
1 PNAs
Vitellogenin
Blood plasma
-------
Jab
le Gene Expression
rory studies, vitellogenin
300
£• 250
'* 200
£ 150
B 100
Jl.
50
0
-50
x
b.
Liver Vg mRNA Levels in Adult Male Fathead Minnows
Exposed to Ethynylestradiol (EE2) in Lab Water
Experiment 1
Lab Water Lab Water
DM SO
10
20
Ethynylestradiol Concentration in ng/L
Moan Vg/18S X 100 Pixel Density + + 1STD
-1STD
-------
lication of Vitellogenin
iicator
Field - Fathead minnows and pearl dace
- Vitellogenin (Vg)
+ Texas - Pecan Creek - Source-biased WWTP
+ University of Kansas - Mesocosms
+ Canada - Whole lake ecosystem study
+ Lake Hartwell - capped sediment study
+ New Mexico - Region 6 - Source-biased WWTP
+ Neuse River Basin - EDC integration into NERL
multimedia model development
-------
On:
wastewater Exposures - Pecan Cr. TX,
ene Expression Levels
CD
X
V)
oo
o>
Vg mRNA levels in Male
Fathead Minnow Livers
D)
>> ^ >>
S S S
(U (S
Q Q
Mean
+ 1STD
-1 STD
CO CO T-
Site Location L = Lab Control, S1 = Site 1 , S3 = Site 3
-------
'te Wastewater cont. Vg Plasma Levels
Vg Plasma Levels in Fathead Minnows
3
0
O)
I 1
D)
E 0
-1
-2
L = I
o
I I
0
A
A
^
A
A
A
Mean
+ 1STD
o
-1 STD
o^cN^^^CN^r^^^^cN^r^^^
O O Q CO CO Q Q Q Q CO CO Q Q Q Q CO CO
_i— icocococo'Tjrcocococojr'^
,ab Control, SI = Site Closest to Effluent, S3 = Farthest from Effluent
-------
Site 1 Vg Protein in Blood of Each Replicate Fathead Minnow
Exposed to Municipal Waste water for 3 Weeks
•i
D , I •• JHhi—[I
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IH^w^-^flflT "ai 3l?^59ai3'?i>'ff*C"J'fl1iI'5'J'fl1i1
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K Aris
• VIG (rng/ni^
Site 1 Vg Expression of Individual Replicate
Fathead Minnow Male Livers Exposed
to Municipal Waste-water for 3 Weeks
0.8 |
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Days ol Exposure af Each Repieaifr ai sue
-------
dian Experimental Lakes Area
\
Winnipeg
Montreal
Minneapolis
-------
Canadian Experimental Lakes Area
-------
EDC Studies- Whole Lake Exposures
Vg Expression in Male Fathead Minnows
Livers of Lake Fish and Cincinnati Fish Exposed to Shipped Samples
1 • ^H
oo ^ n R
i- CO U'°
+ £jz O A
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-------
0.8
0.6
0.4
> 0.2
d) 0.0
JO)
D Control
• DM SO only
D EE2/DMSO
Ohr 8hr 24 hr 4 day 7 day 14 day
rea
Kansas Mesocosm Study - All Trophic Levels
CO
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1.0
0.8
0.6
0.4
0.2
0.0
D Oligotrophic
D Mesotrophic
• Eutrophic
-8h
8hr
24 hr
4 day 7 day 14 day
-------
Single Gene Indicator Studies
- Laboratory - Adult male and larvae fathead
mnnow
Vitellogenin (Vg)
Estrogens (estradiol, ethynylestradiol, DEHP)
CytochromeP450IAI
- PAH's, PCBfs
Metallothionein
etals (Cd, Cu)
-------
Study Themes
* Single stressor / single
known gene
* Multiple stressors /
multiple genes
IDMA
Ino ecu e
Nucleus
Cytoplasm
Gene
activated
Messenger
RNAs
Nucleus
Cytoplasm
Gene
activated
- Messenger
"RNAs
-------
'tujtiple St
DMA
molecule
no
Nucleus I,
Cytoplasm
Gene
activated
- Messenger
RNAs
IDNA
Imolecule
Nucleus
Cytoplasm
Gene
activated
r Messenger
' RNAs
IDNA
Imolecule
Nucleus
Cytoplasm
Gene
activated
Messenger
RNAs
IDNA
Ino ecu e
Nucleus
Cytoplasm
Gene
activated
Messenger
RNAs
IDNA
Nucleus
Cytoplasm
Gene
activated
Imolecule \
Messenger
RNAs
-------
ij)le Gene Expression
tery Studies
* DNA microarrays
- Glass slide with DNA spots
+ Detect thousands of changes in gene expression relative to controls
or other exposures
IDNA
Ino ecu e
Nucleus
Cytoplasm
Gene
activated
Messenger
s
Studied with-
DNA Microarray
-------
fyle Gene Expression
oratory Studies
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Estrogens
Cadmium
Atrazine
PAH's
DNA Microarray
-------
Jt/p/e Stressor Diagnostics
tiole Gene Studies
* Stressors
- EDCs
4- Estrogens
4 Androgens
4 Thyromimetics
4 PAHs
4 Metals
- Pesticides
4 Atrazine
4 Alachlor
- Pharmaceuticals
4 Fluoroquinolones
-------
V Biological Indicators
o from here-
* More exposure questions out there
than the ones we're asking
* Commitment to technology
transfer - demos in your backyard
* Core molecular capabilities are
unsurpassed in ORD
:» Field capabilities are unique
-------
Assessment of Estrogenic Potential
& Bioavailability of PCBs in Lake
Hartwell Water and Sediment
James M. Lazorchak1- David Lattier1, Mark E. Smith2, Barry Wiechman3, Dan
Williams2- Richard C. Brenner4, Victor S. Magar5,
1 USEPA NERL, Cincinnati, OH
2 SoBran Inc. c/o USEPA NERL, Cincinnati, OH
3 PAI, c/o USEPA NERL, Cincinnati, OH
4U.S. EPA, NRMRL, Cincinnati, OH
5Battelle, Columbus, OH
-------
Objectives
• Determine if PCBs in Lake Hartwell water are
estrogenic to male fathead minnows and/or Fry or
if sediments are estrogenic to larval fathead
minnows.
• Determine if Lake Hartwell sediments are toxic to
freshwater amphipods and/or fathead minnow
embryos.
Determine if PCBs in water and sediments of
Lake Hartwell are bioavailable.
-------
Lake Hartwell Site Map
'
Denotes Surface Sample
Transect Location
Denotes Natural Recovery
Transect Locations
•f^t: W
jm
Contaminated
sediment sample
locations:
Transects L & O
Locations
matched USEPA
and natural
recovery
transects
Uncontaminated
water collected
from the Keowee
River
-------
Approach - Water Samples
• 4 Water column samples:
* Bkg = Keowee River
* 3 Transects = T-O, T-L, T-I
• 4 Controls were used:
* 2 positive controls of a 50:50 mixture of
Aroclors 1242 & 1254 at 10 and 100 ng/L in
DMSO
* 2 laboratory water controls of moderately hard
water with and without DMSO at 3.74 wg/L.
-------
Approach - Water Samples (Cont.)
Each water sample was tested by exposing 5
individual 11-13 month old adult male fathead
minnows for 24-hrs.
-------
Approach - Sediment
Nine Sediment samples
* Bkg = Keowee River
* Transects = 3 T-O, 3 T-L, 2 T-I
4 Control sediments
* Sand Control Sediment - Grade 30
* PCB Positive Sand control (10 ng/L of 50:50
Aroclors 1242 and 1254)
* 2 EE2 Positive Sand Controls (10 & 20 ng/L
Ethynylestradiol)
-------
Approach - Sediment (Cont.)
i Each sediment was tested using a total of 160
24- to 48-hr old fathead minnow eggs for 7 days
and collected for Vg analyses after they hatched.
Each sediment was tested using pooled 120
10-day old amphipods for 7days.
-------
Example on How Gene
Expression is Quantified
6
Vg
18S
-------
Pixel Density
Fathead Minnow Fry Data
Name
Volume
RECT-1-Vg
RECT-1-18S
RECT-2-Vg
RECT-2-18S
RECT-3-Vg
RECT-3-18S
RECT-4-Vg
RECT-4-18S
RECT-5-Vg
RECT-5-18S
RECT-6-Vg
RECT-6-18S
15419135.47
61092632.94
14860688.45
67114894.24
11051396.38
59201253.85
17506216.8
52326040.86
15044467.94
41589259.53
13696792.
.53
<"»^
26349086.86
-------
Vg Results in Male Fathead Minnow Livers
Exposed to Lake Hartwell Water Samples
00
a,
~
§>
(/)
c
CD
CD
0.5
0.4
0.3
0.2
0.1
0
-0.1
0
Average
+1 std
-1 std
A
Sample or Site ID
-------
P450 Results in Male Fathead Minnow Liver
Exposed to Lake Hartwell Water Samples
oo
o
un
o
un
CL
>>
c
o
Q
X
E
Average
+1 std
-1 std
A
Sample or Site ID
-------
Survival Results of Amphipods and Fathead
Minnow Eggs in Lake Hartwell Sediments
'>
a—
CO
100
Sample or Site ID
Amphipods
Fathead minnow eggs
-------
Vg Results In Fathead Minnow Eggs Exposed
to Lake Hartwell Sediment Samples
00
ja
g
CD
G
Average
+1 std
-1 std
Sample or Site ID
-------
P450 Results In Fathead Minnow Eggs Exposed
to Lake Hartwell Sediment Samples
oo
O
un
o
un
CL
>>
c
o
Q
"53
x
Sample or Site ID
Average
+1 std
-1 std
-------
Observations - Water
• No toxicity to fathead minnow fry or adults was
found in water samples.
• Vitellogenin gene (Vg) expression was found in
male fathead minnows in all Lake Hartwell water
samples.
No Vg Gene Expression was detected in Fry -
(Exposure may need to be >48hr)
-------
Observations - Water (Cont.)
• Vg Gene expression in Lake Hartwell Water
samples was >10 ng/L < 100 ng/L 50:50
Aroclors 1242/1254.
P450 Gene expression was found in all water
samples and was >100 ng/L PCB.
-------
Observations - Sediment
• Sediment samples from T-O, T-L, and T-I-A
were toxic to fathead minnow eggs - UN-NH3
• Vitellogenin gene expression in FHM eggs
exposed to T-L-B, T-L-C, and T-I-B was higher
than Sand Control or Keowee River sediments.
• Vitellogenin gene expression was > 10 ng/L EE2.
Previous studies have detected Vg expression as
low as 5 ng/L EE2 in FHM eggs and 2 ng/L EE2
in adults.
-------
Observations - Sediment (Cont.)
• P450 expression in Lake Hartwell sediments was
not different than control samples.
P450 gene expression in Fathead Minnows was
not a good indicator of PCB bioavailability.
Working on new genes.
-------
Future Directions & Projects
• Continue EDC studies on Lake Hartwell
* Repeat Vg Water & Sediment tests
* Caged Vg Fish Studies
* Look at Vg Indigenous related species
• National Screening Assessment of 50
Municipal Effluents
• CAFO Screening Assessment in Ohio
-------
An Engineering Approach to
Abatement ofEstrogenic EDCs
in Wastewater
Paul McCauley
-------
Who is involved?
Paul McCauley
Greg Sayles
Barry Austern
Marc Mills
Carolyn Acheson
Eric Kleiner
Richard Brenner
James Lazorchak
Alan Zaffire
George Sorial
Mar Esperanza
Cindy Boardman
Makram Suidan
Chemist
Chemical Engineer
Chemist
Environmental Engineer
Chemical Engineer
Environmental Engineer
Environmental Engineer
Ecotoxicologist
Analytical Chemist
Chemical Engineer
Chemical Engineer
Biologist
Environmental Engineer
U.S. EPA
U.S. EPA
U.S. EPA
U.S. EPA
U.S. EPA
U.S. EPA
U.S. EPA
U.S.EPA
IT Corporation
Univ. Cinti.
Univ. Cinti.
Univ. Cinti.
Univ. Cinti.
-------
The Problem
Publicly Owned Treatment Works (POTW)
Discharges appear to have Estrogenic Effects on
Several Species of Fish //
This effect appears to be mediated through the
estrogen receptor
Compounds Suspected of these Estrogenic effects
include Estrogens and there metabolites andT % /I
Alkylphenols, there ethoxylates and metabolites^
-------
HO
HO
Estrogens and Androgens
Estrone
Estriol
CH
OH
HO
17-p-Estradiol
CH
OH
Testosterone
HO
17-a-Ethynylestradiol
Progesterone
Androstenedione
-------
Alkylphenols
C9H1
4-nonyiphenol (NP)
Me H Me
Me H Me
4-terf/a/y-octylphenol (OP)
(OCH2CH2)n OH
r
R=CgHig, nonytphenol ethoxylates (NPEO)
R=C8H17, octylphenol ethoxylates (OPEO)
-------
Long Term Objectives
1 To determine the fate of Estrogenic EDCs
(including Estrogens, there metabolites;/}
Alkylphenols, there ethoxylate esters and
metabolites) during wastewater treatment
Ml*
2 To fashion an engineering solution tor.
Estrogenic EDC discharge in wastewater>
and sludge. J. *'
• •
-------
Basic Approach
Develop assays for the suspect agents
Construct pilot scale wastewater treatment system
to model metabolic pathways. //
Assay samples from selected and representative
POTWs to determine if the pilot scale system is
modeling POTWs • /,
Optimize pilot scale wastewater treatment system^
for treating estrogenic EDCs i «V j
Make recommendations for improving EDC \ >^
treatment in POTWs
-------
The Proposed
Steroid analysis using solid phase extraction
and GC/mass spectrometry
Nonylphenol ethoxylate analysis using soli
• /
phase extraction and normal phase HPLC.
Fathead minnow using estrogen receptor"
mediated induction of vitellogenin by /
measuring messenger RNA assay (NRML)
Recombinant Yeast assay
r.
t
/
* •*
••
-------
Steroid Analysis
j
Liquid and Solid fractions shall be separated
analyzed separately.
Solids shall be extracted
All fractions will then be concentrated using solid
phase extraction (SPE) Extracts from the SPE will
be derivatized and analyzed by GC Mass */« £
Spectrometry L * ^
Goals for detection thresholds is 1 ng/L for each
steroid.
A -1
-------
Nonylphenol Ethoxylate Analyt
.
i •
* Liquid and Solid fractions shall be separated and
analyzed separately
* Solids shall be extracted
* All fractions will then be concentrated using soli
phase extraction (SPE) Extracts from the SPE will
be analyzed by normal phase HPLC
* Goals for detection thresholds is 50 ng/L for
alkylphenol ethoxylates
-------
Messenger RNA Assay
Supply samples to NERL for mRNA
(vitellogenin) induction analysis in
Fathead Minnow
-------
Recombinant Yeast Assay >
•• Estrogen
H
JAI
• b • •
Estrogen
/c
Receptor
Activated
Receptor
Reporter
Protein
• •
From Routedge and Sumpter, 1996. Environ. Tox and Chem. 15: 241- 248
-------
Pilot scale wastewater
systems
* EDC Pilot Plant with Aerobic Sludge
digestion
* EDC Pilot Plant with Anaerobic Sludgd g
digestion j '/j Jj
-------
EDC PILOT PLANT FLOWSHEET WITH AEROBIC SLUDGE
DIGESTION
PRIMARY
CLARIFIER
AERATION TANK
SECONDARY
CLARIFIER
RAW
WASTEWATER
PRIMARY
EFFLUENT
»o o oo - oo
. . •"'o.ooo
oo
= «,0
00* o
n nnn n n nnn n
MIXED
LIQUOR
RETURN ACTIVATED SLUDGE
PRIMARY SLUDGE
WASTE ACTIVATED SLUDGE
MIXED SLUDGE
O o 80
-------
EDC PILOT PLANT FLOWSHEET WITH ANAEROBIC SLUDGE
ION
PRIMARY
CLARIFIER
AERATION TANK
RAW
WASTEWATER
PRIMARY
EFFLUENT
PRIMARY SLUDGE
FILTRATE OR CENTRATE, E.G.
o o °
o
°«,o * o
"tf. O
O o O
****** ""O ~ "* n W I
« o °o0 o* o ' oo - ,
nnnrrn n nnn n n nnn n
. o » o
° o. o
MIXED
LIQUOR
RETURN ACTIVATED SLUDGE
WASTE ACTIVATED SLUDGE
MIXED SLUDGE
METHANE GAS
DIGESTED SLUDGE
SLUDGE
DEWATERING DEVICE
DEWATERED SLUDGE
SECONDARY
CLARIFIER
-------
EDC PILOT PLANT FLOWSHEET WITH AEROBIC SLUDGE
DIGESTION
PRIMARY
CLARIFIER
AERATION TANK
SECONDARY
CLARIFIER
RAW
WASTEWATER
PRIMARY
EFFLUENT
« »a">o00 ,
oa
= «,o
nnnrfn n nnn n n nn?i n
MIXED
LIQUOR
RETURN ACTIVATED SLUDGE
PRIMARY SLUDGE
WASTE ACTIVATED SLUDGE
MIXED SLUDGE
,00 o
80 o o
o.o
-------
EDC PILOT PLANT FLOWSHEET WITH ANAEROBIC SLUDGE
ION
PRIMARY
CLARIFIER
AERATION TANK
RAW
WASTEWATER
PRIMARY
EFFLUENT
OO°
O
°«,o * o
a a o —
• ••; °o. o*.° :.•••.
nnnrfn n nnn n n nnn n
MIXED
LIQUOR
RETURN ACTIVATED SLUDGE
PRIMARY SLUDGE
WASTE ACTIVATED SLUDGE
MIXED SLUDGE
METHANE GAS
FIL TRA TE OR CENTRA TE, E. G.
DIGESTED SLUDGE
SLUDGE
DEWATERING DEVICE
DEWATERED SLUDGE
SECONDARY
CLARIFIER
-------
EDC Pilot
Plant
-------
Aerobic Tanks
-------
Secondary Clarifier
-------
Anaerobic Digester
-------
Make recommendations to
Improve Treatment of Estrogen)
EDCs
* Analyze results of pilot scale systems ai
modified pilot systems
* Compare results to field results (POTWs)
* Report the generated data and make final ^
recommendations for improved Estrogenic/ /
EDC removal. Published as either a journal
Article or EPA report
. ••
• •
-------
Alkylphenols and Ethoxylates
OH
R
Alkylphenol
R
Alkylphenol polyethoxylate
R -O(CH CH O)nCH COOH
O p ' I i O
Alkylphenol polyethoxycarboxylate
R = C H (octyl)
8 17 l
= C H (nonyl)
9 19V y/
R is usually branched
-------
A
Metabolic fate of Alkylphenolics
-O-(CH2-CH2-O-)nH
NPnEO
NP(n-1)EO
aerobic
R—({ V-O-CH2-CH2-O-CH2-COOH
NP2EC
\
O-CH2-COOH
NP1EC
anaerobic
-OH
NP
., H
anaerobic
0-(CH2-CH2-O-)2H
NP2EO
I
R
O-CH2-CH2-OH
NP1EO
anaerobic
-------
Risk Management Research:
Improving Environmental
Decisions
Workshop on Effective Risk Management
of Endocrine Disrupting Chemicals
Cincinnati, Ohio
January 29, 2002
Hugh W. McKinnon, M.D., M.P.H.
Associate Director for Health
National Risk Management Research Laboratory
U.S. Environmental Protection Agency
US EPA OIIICB of R»seaich and Devslopnmnt
-------
Risk Assessment/Risk Management Paradigm
Risk Assessment
Dose-Response
Assessment
Hazard
Identification
Exposure
Assessment
Risk
Characterizatio
Risk Management
Decision
Non-Risk
Analyses
Social
Concerns
International
Relations
Control
Options
""V
Public \
Perception )
Economics,
Trade
Risk Management!
-------
Historical Perspective
Focused on Uncertainty
"The dominant analytic difficulty [in decision-
making based on risk assessments] is pervasive
uncertainty...There is often great uncertainty in
estimates of the types, probability and magnitude
of health effects associated with a chemical
agent, of the economic impacts of a proposed
regulatory action, and of the extent of current and
possible human exposures."
"Risk Assessment in the Federal
Government" (National Research
Council, 1983)
-------
Source-Exposure-Dose-Effect Continuum
SOURCE /STRESSOR
FORMATION
Chemical
Physical
Microbial
Magnitude
Duration
Timing
DISEASE
TRANSPORT /
TRANSFORMATION
Dispersion
Kinetics
Thermodynam ics
Distributions
Meteorology
ALTERED STRUCTURE /
FUNCTION
ENVIRONMENTAL
CHARACTERIZATION
Air
Water
Diet
Soil & dust
Activity
Patterns
Cancer
Asthm a
Infertility
efc.
EARLY BIOLOGICAL I
EFFECT I
Edema
Arrhythm ia
Enzym uria
Necrosis
efc.
EXPOSURE
DOSE
Molecular
Biochemical
Cellular
Organ
Organism
Pathway
Route
Duration
Frequency
Magnitude
Individual
Community
Population
Absorbed
Target
Internal
Biologically Effective
Statistical Profile
Reference Population
Susceptible Individual
Susceptible Subpopulations
Population Distributions
-------
Recent Emphasis Focuses on
the Use of Mechanistic Data
"The quality of risk analysis will improve as the
quality of input improves. As we learn more
about biology, chemistry, physics, and
demography, we can make progressively better
assessments of the risks involved. Risk
assessment evolves continually, with re-
evaluation as new models and data become
available."
"Science and Judgment in Risk
Assessment" (National Research
Council, 1994)
-------
Risk
Management/
Risk Assessment
Paradigm
Identification of Future Problem, Initiating
Event or
Public Policy Mandate
Risk Management
Formulate the Problem
X
Dose-Response
Assessment
Define Risk Management
Objectives
Risk Assessment
Hazard
Identification
Risk
Characterization
Identify and Evaluate
Risk Management Options
Exposure
Assessment
Risk Management
Decision
Develop Compliance
Assurance Models and Methods
Implement Option(s)
Develop Measures of
Environmental and
Public Health
Improvement
Monitor
Environmental
and Public Health
Improvement
' Public Health
Considerations
1 Statutory and Legal
Consideration
1 Social Factors
1 Economic Factors
1 Political
Considerations
Reduced Environmental
and/or
Public Health Risk
-------
ENVIRONMENTAL RISK
Fields of Analysis
Risk Assessment
• Nature of effects
• Potency of agent
• Exposure
• Population at risk
•*> Average risk
•*> High-end risk
•*> Sensitive groups
• Uncertainties of science
• Uncertainties of analysis
Identify
Describe
Measure
Risk Management
Social importance of risk
De minimis or acceptable risk
Reduce/not reduce risk
Stringency of reduction
Economics
Priority of concern
Legislative mandates
Legal issues
Risk perception
Evaluate
Decide
Implement
-------
OBSERVATIONS ON RISK-BASED
DECISION MAKING IN EPA
•Risk assessment and risk management within
EPA are rarely if ever separated by sharp lines.
Rather, decision-making is an interative process
that considers available data and information on
both the risks in question and the remedies
available to mitigate the risks.
•The uncertainties, costs, commercial viability,
and feasibilities attendant to available remedies
influence the rigor demanded from all
assessments required to define the problem and
assess the risks.
-------
OBSERVATIONS ON RISK-BASED
DECISION MAKING IN EPA (cont.)
• Where legal or political mandates are clear and
precedents are unambiguous, risk management
decisions tolerate high degrees of uncertainty in
both risk assessments and risk management
remedies, especially when costs are modest.
• Similarly, as the Agency moves toward
advocacy (e.g., incentives, community-based
programs, emission-trading, National action
Plans) as a means to foster better decision-
making and when costs of decisions are high,
uncertainties in both risk assessment and risk
management must be reduced.
-------
INFLUENCE OF SCIENTIFIC UNCERTAINTY ON TIME
PHASING OF RISK MANAGEMENT DECISIONS
£<
t
a
z ¥>
ID a:
Risk Level that Once
Accepted Enables Risk
tent Decisions
lower uncertainty
time
Risk Level Accepted and
Risk Management Decisions
Made
Risk Reduction
Acceptable Risk Level
WHAT ABOUT UNCERTAINTIES IN RISK MANAGEMENT?
-------
I INFLUENCE OF UNCERTAINTY ON TIME PHASING
t FOR RISK ASSESSMENT AND RISK MANAGEMENT:
N
3 IDEAL CASE
Risk Level that Once
Accepted Enables Risk
lent Decisions
low uncertainty
LJJ
LJJ
<
Z
<
* Z
(/) —
&• Q_
Z O
Risk Level Accepted and
Risk Management Decisions
Made
Risk Reduction
Acceptable Risk Level
IDEAL CASE-- RISK ASSESSMENT AND RISK MANAGEMENT
EVALUATION ARE COORDINATED
-------
N
15 S
m
^
m
3|»
ssl
Us
INFLUENCE OF SCIENTIFIC UNCERTAINTY ON
RISK ASSESSMENT AND RISK MANAGEMENT:
IMPACT OF DELA YS
(sk Level that Once Accepted
lers Risk Management Decision
Risk Management
Decisions Made
"Effective" RM
Implementation
Risk
Reduction
DELAYED RISK MANAGEMENT CHARACTERIZATION LEADS TO
INCREASED RISKS
-------
INFLUENCE OF SCIENTIFIC UNCERTAINTY ON RISK
ASSESSMENT AND RISK MANAGEMENT: IMPACT OF
ACCELERATED RISK MANAGEMENT CHARACTERIZATION
N
O
o
CO
£
LJJ
^
LJJ
^ o
UJ CO =
sk Level that Once Accepted
TrKKiers Risk Management Decision
Early Risk
Management
Decisions Made
risk reduction based on
ailability of low-cost,
effective options
Risk reduction based on
acceptable uncertainty in
risk characterization &
risk management
LU
>
LiJ
_l
^
CO
ACCELERATED RISK MANAGEMENT CHARACTERIZATION LEADS TO
REDUCED RISKS
-------
Hazard ID
RISK ASSESSMENT
Risk Characterization
Dose Response
Exposure Assessment
V
TIME TP1 TP2
TP3
TP4
TP5
TP6
Source
Characterization
Risk Management Options
Cost and Effectiveness
Assessment
Risk Management Options ID
= TIME PERIOD
RISK MANAGEMENT EVALUATION
TIME PHASING FOR RISK ASSESSMENT AND RISK MANAGEMENT
CHARACTERIZATION TO ACHIEVE OPTIMAL RISK REDUCTION
-------
RISK MANAGEMENT EVALUATION
DEFINITION:
An analysis of:
- sources of potential, perceived, or actual risk,
- RM options for preventing or reducing risk,
(primarily technical, e.g., control, P2)
and
- availability, costs and effectiveness
of the identified RM options.
-------
RISK MANAGEMENT EVALUATION
GOAL OF RISK MANAGEMENT
EVALUATION RESEARCH:
Reduce uncertainty in Risk Management for EPA
and other public and private sector organizations
and provide cost-effective RM options (both
technical and non-technical) for preventing,
reducing, or adapting to current and emerging
high risk problems.
-------
RISK ASSESSMENT
Statutory and Legal
Considerations
Dose Response
Assessment
Public Health
Social
Considerations Considerations
Hazard ID
Exposure
Assessment
Source
Characterization
Risk
Characterization
Risk Management
Decision
Risk
Management
Options
\
Economic
Factors
political
Risk Management
Options Identification
Considerations
RISK
MANAGEMENT
EVALUATION
Cost and Effectiveness
Assessment
RISK
MANAGEMENT
-------
RISK MANAGEMENT EVALUATION
Source Characterization
• identification-who, what, where
• source strengths-how much, what
form
• timing-changes overtime
• emission factors-estimation
w
:
Risk Management Options
Identification
• pollution prevention
• source control technologi
• management practices
remediation
restoration
adaptation
L
RISK
MANAGEMENT
OPTIONS
Effectiveness and Cost Assessment
• verification status-commercially available?
• known effectiveness-data
• estimated effectiveness-engineering judgment
• reliability estimates-failure modes & rates
• capital costs
• O&M costs
-------
PILOT RISK MANAGEMENT
EVALUATIONS UNDERWAY IN
ORD
Endocrine Disrupters
Arsenic In Drinking Water
o CAFOs
o Participate Matter (PM2 5)
-------
Excerpt from "Risk Management Research: Improving Environmental Decisions"
Hugh McKinnon, NRMRL
EDC Workshop January 29, 2002
In 1995, the Office of Research and Development in EPA, re-organized into our current configuration,
creating three national laboratories and two national centers. At that time, we became the National Risk
Management Research Laboratory, and remembering that our predecessors had been primarily
engineering and technology development laboratories, we decided to take a look at the risk
management side of the process. We felt the risk assessment side had received a fair amount of scrutiny
and study, but the risk management side had not. So we decided to try to draw a better depiction of
the risk management process, and among the senior management in ORD collectively came up with this
diagram. I don't know who to credit, but someone alertly observed that the risk assessment process is
at the core of this, fits into the whole matter of making risk management decisions, and came up with
this diagram that looks like a cell, with the risk assessment as the nucleus. Out here, in very simplistic
form, are laid out some of the major steps in making a risk management decision, implementing that
decision, and looking at what the consequences are.
There is a great variety of risk assessment, risk management, and other considerations, that come into
play here. So as we consider decision making, keep in mind that we are the research and development
arm of EPA-the people who try to provide scientific and technical support to the rest of the Agency.
We are not the risk managers, we're not those who, in the Agency, are setting the regulatory standards,
nor those at the state and local levels who are actually having to make decisions and implement them.
But, in looking at the process, we came up with some observations. One is that risk assessment/risk
management are rarely sharply separated. Decision making tends to be iterative and it considers all the
data available. Secondly, there are uncertainties, as well as costs and other considerations, that play
into risk management and risk assessment, and those have not really been looked at and defined so well
in decision making. Third, we observed that, where legal or political mandates are clear and where we
have unambiguous precedents, risk management decisions will tolerate a great deal of uncertainty in
both the risk assessment and remedies, especially if the costs are not high. If the costs are high
however, or if the precedents are not so clear, then the uncertainty question comes into play and people
begin to demand a great deal of reduction in uncertainty.
Finally, as our Agency moves toward less of a command-and-control approach and more toward what
we've called an advocacy approach, things like incentives, community based programs, consideration
of trading programs, action plans that are so broadly defined at the national level but left for local
implementation and decision, and when the costs of decisions are high, then uncertainties in both risk
assessment and risk management need to be reduced. That, in a nutshell, is what we've tried to do
since the formation of the Risk Management Research Lab. We're continuing to try to do that,
continuing to try to bring increased definition of our risk management understanding and programs that
broaden the scope of discipline that those might include, and that will continue.
Along the way, Lee Mulkey came up with the idea of something we're calling a risk management
-------
evaluation. The risk management evaluation is a summary of what we know about risk management
options at the time a decision needs to be made. It would include an analysis of sources of risks, the
risk management options that exist for preventing or reducing those risks, and the availability, cost, and
effectiveness of those options. Again it was not our intent to second guess or re-hash the complex risk
assessments that often accompany Agency decisions. It was rather to look at the other side of the
paradigm, if you will, at the risk management side. In order to do that, we might indeed have to provide
a capsule summary of what we know about risks. So the goal of these evaluations and the research to
support them is to reduce uncertainty in risk management for EPA and others, in order to provide them
with cost effective risk management options. These might be technical or non-technical, for preventing,
reducing or adapting to current and emerging high-risk problems. So we came up with this diagram to
encapsulate what we were saying about the risk management evaluation. You will recognize this is the
risk assessment side of the paradigm, and this is the risk management side and we've added a third oval
to summarize what's included in a risk management evaluation. Hopefully, the risk management
evaluation complements the risk assessment and plays into effective risk management decision making.
-------
Development of Chemical Methods to
Characterize Exposure to EDCs in the
Neuse River Basin
M. Medina-Vera, S. Harper, L. Wright, E. Coppedge,
S. Lumpkin
US EPA/ORD/NERL/HEASD
G. Ferrell
USGS/Raleigh
EnSenll PniKfcn OS*8) US EPA Office of
Anancv -^^ M7 Research & Development
uses
for a changing world
-------
EPA Near Laboratory Ecological Research
Areas (NLERA)
Cincinnati, OH- Little Miami River Basin
Las Vegas, NV-Colorado River Basin
Athens, GA- Savannah River Basin
Research Triangle Park, NC- Neuse River Basin
EnSenll Protect™ A!*1*) US EPA °"** °f
AnEncy -^ M7 Research & Development
uses
for a changing world
-------
NORTH CAROLINA RIVER BASINS
f&SQUOTA/'K
fRENCH SfKsAD
UTTLE
T£N/'f£S$££
SCALE 1:5,000,000
Q HD 2{ID MILES
Map Prepared by the N.C. Geographk Information & Analysis * 115 Hillsborough Street* Raleigh, NC 27603 * (919)733-2090
US EPA Office of
Research & Development
uses
ftv a changing world
-------
Potential Sources of Toxic Elements in the Neuse River Basin
North Fiat River
-•• . * Flat River
•*.-« • r
Contenlnea Creek
•
I
; .-* -.
-• • -• ' 'V- .".
Neuse River
..
Map F
venation JAcfrjcv8l/ALK& Facility iubsystc^t
'L'u(ii|m:ti^iLiivt Ln-'-irL'tnil^nLiJ, .-i-^-.-jKHiie, dicri|«;:ixa.l>jn mid LiubiliLy
Permit (^^mplianLe Jiyvt&m (Water Di;eh;irir.<* Perm't*.)
Sti^blaiioc Di&pcaal SJu-a
CWLT iiy.iLrinji - LJIIK: AppEicataim
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• Cyanic
Hoars
• Pauliry
.X: 1iiidC\!',er
3OSK9M- Cbard:i<>.1u:loii ttixul Ari?lysl;
>Wflrr» T>wijl« "Pjrtr K"iirn l"nni>lini
-------
Methods Development
Organic compounds
Alkylphenol Polyethoxylates (APEOs)
Antibiotics
Pesticides
Inorganic compounds
Toxic Elements
- Focus: As, Cd, Cu, Mn, Pb, Sn
EnSenll PniKfcn OS*8) US EPA Office of
Anancv -^^ M7 Research & Development
uses
for a changing world
-------
Alkylphenol Polyethoxylate Research
Why are they important?
- Breakdown products are considered more toxic than parent compounds.
- Persistent in rivers, sediments, and groundwater.
- Bioaccumulated by fish and birds.
- Considered as EDC mimickers.
Sources/Uses (Examples)
- Manufacturing: pulp and paper, textile, plastic & elastomer, leather.
- Household, industrial, and institutional cleaning products, spermicides in
contraceptive gels, jellies, creams.
- Agriculture.
EnSeny Protect™ A!*1*) US EPA °"** °f
AnEncy -^ M7 Research & Development
uses
for a changing world
-------
Degradation Mechanism of APEOs
Ha iHe
" R=C^H19.non^
y\,
ft LH= Jm Ha R is usually tranched
pDlyethoxylate
(APnEO, n =m+ 1)
Alk ylf±enoxy acetic acid
(AP1EQ
fiolyethoxylate
AP(n-1)"EO
:ive shortening of ethoAviate chain
R AlKylp^ienol monoethoxylate
(AP1EO)
&EPA
Uniled Slater
-I vii ,• i i • i. 'I I •, i,;,', , i
OH
\
Alkyiphenol (AP)
Ring cleavage, o^dation ofalhyl chain
US EPA Office of
Research & Development
uses
for a changing world
-------
Organic Compounds Sampling Methods
Water: Amber collection bottles dipped at least 5" deep upstream side
of the boat.
Sediment: Grab samples (1/25 m2) stainless steel coated with Kynar.
Fish: USGS method adaptation of Report 93-104.
EnSenll Protects OS*8) US EPA Office of
Anancv -^^ M7 Research & Development
USGS
for a changing world
-------
Analytical Methods
Isolation/Extraction Methods
- Solid Phase Extraction (SPE): Water, Sewage.
- Accelerated Solvent Extraction (ASE): Sediment
Chromatographic Methods
• HPLC Reverse Phase: ODS Hypersyl, 100% H2O/0.005 KH2(PO4) to 100%
CH3CN, 30 minutes, 0.250 mL/min flowrate. Reverse Phase: 100% Methanol
Isocratic, 35 minutes.
• HPLC Normal Phase: NH2 column, gradient 10% Isopropanol, 90% Hexane to
100% Isopropanol, 0% Hexane, 35 minutes.
EnSenll PniKfcn OS*8) US EPA Office of
Anancv -^^ M7 Research & Development
uses
for a changing world
-------
Normal Phase Chromatography
Print of window 38t Current Chromatogram(s)
Current Chromatogram(s)
WDnr5lg.22Z.20 RH-4HJ.i
mAU
Instrument 1 3/21/2001 10i03i43 AM LWright
Page 1 of 1
&EPA
United Slaes
US EPA Office of
Research & Development
uses
for a changing world
-------
Print of window 80: MS Spectrum
MS Spectrum
Mass
Spectrum
100-
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Icms 3/23/01 10:26:02 AM ferrell
^* ^"^%A United Slates ^T^at us FPA CiHirpnf
Jttj ^H brXl Envii tnnjf n'.al Protectnn f • •! jl
^|4r ^H 1 /"\ Agency *B^ fl7 Research & Development
i
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Page 1 of 1
^^•rusGS
-------
Toxic Elements Research
Why are they important?
- Persistent, bioaccumulates, toxic.
- Impacts on reproductive, immune, neuro-behavioral and endocrine system
functions in developing and mature organisms.
Sources/Uses (Examples)
- Used as growth promoters for chickens.
- Herbicides, wood preservatives, semiconductor manufacturing, petroleum
refining, mining and smelting operations.
- Medical and municipal waste incinerators, utility boilers, paints, fish.
EnSeny Protect™ A!*1*) US EPA °"** °f
AnEncy -^ M7 Research & Development
uses
for a changing world
-------
Toxic Elements Methods
Example: Surface Water Sampling
- EPA Method 1669, "Sampling Ambient Water for Trace Metals at EPA
Quality Criteria Levels".
Analytical
- Confirmation: Absorption Spectroscopy.
- Modified EPA Method 1639, "Determination of Trace Elements in
Ambient Waters by Stabilized Temperature Graphite Furnace Atomic
Absorption".
- Modified EPA Method 200.9 "Determination of Trace Elements by
Stabilized Temperature Graphite Furnace Atomic Absorption".
EnSeny Protect OS*8) US EPA Office of
^gEncy -^^ M7 Research & Development
uses
for a changing world
-------
Toxic Elements Results-1
NFRR: North Flat
River Reach
FRR: Flat River
Reach
CCR: Contentnea
Creek Reach
NRR: Neuse River
Reach
Mn Concentration Across
Neuse River Basin
35.40%
NFRR
FRR
CCR
NRR
33.22%
&EPA
United •Slates
Environmental I •, i,; J,, i
US EPA Office of
Research & Development
uses
for a changing world
-------
Toxic Elements Results-2
NFRR: North Flat
River Reach
FRR: Flat River Reach
CCR: Contentnea
Creek Reach
NRR: Neuse River
Reach
Sn Concentration Across the
Neuse River Basin
23.90%
21.20%
27.W°A
NFRR
FRR
CCR
NRR
27.90%
&EPA
United •Slates
Environmental Procter!
US EPA Office of
Research & Development
uses
for a changing world
-------
Future Directions
Continue developmental work on field and analytical methods.
Provide environmental baselines for targeted compounds/elements at
established sites in the Neuse River Basin.
Continue research with the Regional Offices on APEOs.
EnSenll Protects OS*8) US EPA Office of
Anancv -^^ M7 Research & Development
uses
for a changing world
-------
Acknowledgments
EPA/ORD/NERL/ESD: Landscape Characterization Branch, RTF,
NC: Neuse River Landscape data & coverages.
EPA/ORD/NERL/HEASD/IO-RTP, NC : Peter Knudsen-graphics
support.
EnSeTy PniKfcn OrB^l US EPA Office of
Anancv -^^ M7 Research & Development
uses
Aw s changing world
-------
Excerpt from "Welcome from the National Risk Management Research Laboratory"
Lee Mulkey, NRMRL
EDC Workshop January 29, 2002
As I look at this program, it strikes me that there are three things that are somewhat different about this
topic and about the way we organized the work. First of all, as a way of introducing the idea, the
budget category in our organization that we typically identified EDCs with is something we call
Emerging Risks. That's an important idea because, as you know, EPA typically responds and reacts to
issues that come up. We looked at a river that burned in Cleveland years ago, Superfund was spun up
after incidents like Love Canal and Times Beach, and a whole new industry on bioremediation was
generated from an oil tanker rupture in Alaska. Here is a chance where EPA has had the foresight to
consider what may be a real risk that requires either new policy development or new initiatives in
implementing or calibrating our regulatory programs dealing with EDCs.
There are two other things that come to mind that I want to share with you before we get down to the
real work. One is that one surely needs to worry about the footprint of EDCs. Now let me explain a bit
what I mean by this. Ecologists typically talk about ecological footprints of human activities and the
matrix they use. As you may know, these are the per-capita appropriations and areal extent of the
natural environment that we humans appropriate for those goods and services we get. One of the
important services that we appropriate from the environment is its assimilative capacity for our waste
and our residuals. Now one surely must wonder-what's the footprint of EDCs? We may not think
about EDCs as a waste, but you're going to hear in this program about the sources and the sinks, and
although we'll continue to depend upon the environment to assimilate EDC residuals, the extent of our
use and the areal extent of releases are still quite large. So surely, one must appropriately wonder
about the risks and try to get a handle on them.
Now the third thing that this idea of an Emerging Risk gives us the opportunity to do is to engage in
what we might phrase 'integrated science.' We are welcoming you on behalf of the Risk Management
Research Laboratory, but this topic reaches across all of the research elements within EPA. And, in a
parallel fashion, we are trying to develop our knowledge base by asking: what are the risks of EDCs,
what are the effects, what's the extent of the exposure, what are the probabilities of the scenarios and
ways and places where intervention might be appropriate, what are the appropriate options for
managing the risks once they're identified and where and how much, and at what cost, and toward
what benefits? This is a good model, I think, for how we ought to think about integrating our science
and our research in order to inform the policy making steps that may be required at EPA and the
federal government, at the state agencies and, through interaction with our colleagues, at the
international community.
-------
Risk Management of Endocrine
Disrupting Chemicals (EDCs) in
^ Drinking Water
Frederick W. Pontius, P.E
Gary Amy, Ph.D.
January 29-30, 2002 USEPA EDC Workshop
-------
Objective
Present a water utility perspective on
EDC risk management
• Are current regulations adequate?
• Should we be concerned about
unregulated EDCs/pharmaceuticals?
• What can water utilities do to manage risk?
• What are the critical research questions?
January 29-30, 2002 USEPA EDC Workshop
-------
Acknowledgements
Gretchen Watson, Univ. of Colorado
• SEM images
Prof. Scott Summers, Univ. of Colorado
AWWA Research Foundation
. Workshop held April 19-21, 2000
January 29-30, 2002 USEPA EDC Workshop
-------
Are Current Regulations
Adequate?
The Drinking Water Context
Existing and Future Regulations
SDWA Authority
January 29-30, 2002 USEPA EDC Workshop
-------
The Drinking Water Context
'Endocrine disruption' -^ an effect...
Disruption of the endocrine system is
just one of many health effects that
could be associated with a drinking
water contaminant
• Inorganic; Organic (Natural; Synthetic)
• Cancer; Noncancer
January 29-30, 2002 USEPA EDC Workshop
-------
Drinking Water Context (Cont.)
• Drinking water quality regulated by
USEPA under the Safe Drinking Water
Act (SDWA)
• Bottled water quality regulated by the
Food and Drug Administration (FDA)
• Other environmental exposures
regulated by other laws
January 29-30, 2002 USEPA EDC Workshop
-------
Contaminants Regulated
Stage 2 DBPR/LT2 m n CCL 2
-o/> Staqel DBPR/IESWTR. /
1 £\J
rr\ ^ rtrt
+$ 1UU
a
2 on
£H ****
• i-H
W C/\
ctf tlU
O AH
*IU
^ TTHMs
ori~^
^U
NIPDWRs^
\ As Rn
Phase V \ \ ^
\
Phase II
SWTR/TCR ^r
Dhase 1 \
-\ \ \
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n
1
**
i
i i
' <
-
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f
f
i
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i
<
, / '
i i i i i i i i i i i i i i i
D Current • Future
(A
CO
re
ea
5
2C
i i i i i i
I C«
(Assumes 5
contaminants
regulated from
each CCL every
5 years starting
January 29-30, 2002
USEPA EDC Workshop
-------
SDWA Authority
• USEPA regulates contaminants that...
• Occur or are likely to occur in drinking
water
• May have adverse effects on human health
• Have a meaningful opportunity for risk
reduction
• Specific contaminants regulated
- MCLG •*• MCL; BAT; Benefits/Costs; Risks
January 29-30, 2002 USEPA EDC Workshop 8
-------
Should we be Concerned
About Unregulated
EDCs/Pharmaceuticals?
Occurrence
Health Risks
Treatment Effectiveness
January 29-30, 2002 USEPA EDC Workshop
-------
Occurrence and Health Risks
Sometimes we find'em, most(?) of the
time we don't
• If detected, then we have evidence.
• If not detected, then ?
• Many types of chemicals, configurations
Potential health effects may be
significant, exposure and health risk
uncertain
January 29-30, 2002 USEPA EDC Workshop 10
-------
EDCs/Pharmaceuticals
in Source Waters
• Inorganic (natural; discharges)
• Organic (natural; synthetic)
• Specific contaminants
• effluent dominated streams/lakes
• Total Organic Carbon (TOC)
• Dissolved organic matter (DOC)
• Natural organic matter (NOM)
• Effluent organic matter (EfOM)
• Soluble Microbial Products (SMPs)
• Algal organic matter (AIOM)
January 29-30, 2002 USEPA EDC Workshop 11
-------
Caffeine
C8H10N402
MW = 194.19
MP = 238 C
Cwsat= 21,600mg/L
Log Kow = -0.07
P° = 15 mm Hg
pKa = 10.4
H = 1.9x10-19atm-
m3/mole
o
N
January 29-30, 2002
USEPA EDC Workshop
12
-------
1 Progesterone
30^2
MW = 314.47
MP = 121 C
Cwsat =8.81 mg/L
Log Kow =3.87
P° = 1.3 x 10-6 mm
Hg
H = 6.49x10-8atm-
m3/mole
January 29-30, 2002
USEPA EDC Workshop
13
-------
beta-Estradiol
MW = 272.39
MP = 178. 5 C
Cwsat= 3.6 mg/L
Log Kow = 4.01
P° = 1.26x 10-8 mm
Hg
H = 3.64X10-11
atm-m3/mole
January 29-30, 2002
USEPA EDC Workshop
14
-------
Allopurinol
C5H4N40
MW = 136.11
MP = 350 C
Cwsat = 569 mg/L
Log Kow = -0.55
P° = 4.73 x 10-6 mm
Hg
H = 2.03 x10-14
atm-m3/mole
January 29-30, 2002
USEPA EDC Workshop
15
-------
I Erythromycin
MW = 733.95
MP = 191 C
Cwsat= 1.4 mg/L
Log Kow = 3.06
P° = 2.28x10-27
mm Hg
H = 5.42 x 10-29
atm-m3/mole
January 29-30, 2002
USEPA EDC Workshop
16
-------
EDC/Pharmaceuticals
1 ^
I 3
m -I
I U
o
O)
o
n
U H
(
MW vs Log Kow
•
wjj!$:* *
) * 200 * ^400* 600 800 10
MW
* Pharm • EDCs - reg EDCs - Unreg VOCs - Reg
00
January 29-30, 2002
USEPA EDC Workshop
17
-------
EDCs/Pharmaceuticals
MWvsH
mn -,
I UU
Onni
.UU I /
1 P Oft
I t-Uo
11 p 1^
I C- I o
1 F 1ft
I C~ I O
1 P 9^
I t-ZO
1 P 9ft
1 * • «r '
) * ^Q? t^1" ^00 600" 800
10
*£$&& *
* \* *
* , **
^ ^ *
I C-ZO
MW
* Pharm • EDCs - Reg EDCs - Unreg VOCs - Reg
00
January 29-30, 2002
USEPA EDC Workshop
18
-------
What Can Water Utilities Do
To Manage Risk?
The water utility perspective
Optimize using current
regulatory control strategies
Due diligence
January 29-30, 2002 USEPA EDC Workshop 19
-------
Knowledge 4=** Consumer
Utility
EPA Scientific Research/
Technical Studies
V
•f
Risk Assessment
Dose-Respons
Characterization
Characterization
MCLG/MCL
Consumer
Source water perceptioi
& Treatment^
Exposure
Characterization
Animal Toxicology
Clinical Studies
Epidemiology
Cell/Tissue
Experiments
Computational
Methods
Monitoring/
Surveillance
•Treatment
•Source protection
teste disposal
Collaboration
^
Other Federal Agencies
States/Local Agencies
Academia
Industry
Public Interest/Environmental Groups
Water Utility
Risk Management
EPA Regulations ^w
State Regulations
Leadership
Compliance status
Customer attitudes
Ability to pay
Politics
Control Options
-capital const.
-operational
External
Research &
Assessment
Decisions
-------
EDC/Pharmaceutical Control
J.M. Symons, 1994
January 29-30, 2002
USEPA EDC Workshop
21
-------
Enhanced Coagulation (EC)
• SW - EC effective for NOM humic
fraction
. SUVA = UV254/DOC (L/mg-m)
• Good indicator of water's humic content
• Non-humics less amenable to enhance
coagulation
• SUVA
-------
Surface Water TOC
IE. Treatment Rant Influent D/DBP Questions
Question 1. What are the levels of DBF precursors?
Mean Influent TOC by State Plant Means of 12 Months
Surface Water Plants (n= 353)
C1A2DMS.V3 GOQ3JE3.SAS
January 29-30, 2002
USEPA EDC Workshop
23
-------
Groundwater TOC
IE. Treatment Rant Influent D/DEP Questions
Question 1. What are the levels of DBF precursors?
Mean Influent TOC by State Plant Means of 12 Months
Ground Water Plants (n= 331)
Mean TOC [ppm]
D No Data
> - 2
> =3
>=4
- 2
— 3
- 4
C1A3DMS.V3 GQQ3JE3.SAS
January 29-30, 2002
USEPA EDC Workshop
24
-------
Particle Removal Processes
Size,
Approximate
Molecular
Weight
Relative
Size of
Various
Materials in
Water
Separation
Process
Ionic
Molecular
Macromolecular
Microparticle
Macroparticle
0.001 0.01 01 1.0 10 100 1.000
100 200 1,000 10,000 20,000 100,000 500,000
Viruses
Aqueous Salts
Metal
Ions
Humic Acids
Bacteria
Algae
Clays
Cysts
Sill
Sand
Conventional Filtration Processes
Microfillration
Ultrafiltration
RO
ED/EDR
January 29-30, 2002
USEPA EDC Workshop
25
-------
Microfiltration
January 29-30, 2002
USEPA EDC Workshop
26
-------
Ultrafiltration
January 29-30, 2002
USEPA EDC Workshop
27
-------
Toxic Tort Action
Lawsuit claims trihalomethanes were cause
154 women sue Chesapeake, Va.
saying water caused miscarriages
CHESAPEAKE, Va. (AP)—
An additional 129 women
have filed lawsuits claiming
the city's drinking water
caused their miscarriages,
bringing the total number of
plaintiffs to 154. Thewomen are
seeking more than $1 billion.
The lawsuits allege that the
city knowingly and/or negli-
gently poisoned the women
and their fetuses when the
women drank, showered,
bathed in, or used city water
containing trihalomethanes,
orTHM.
more lawsuits are expected.
City officials have disputed
the allegations and claimed
that Chesapeake has stayed
within the federal guidelines
for safe drinking water.
The city also contends it
cannot be sued because the
lawsuits were filed after a two-
year statute of limitations had
expired, and that the city is
protected by sovereign immu-
nity. City officials have asked
that any court proceedings be
moved to another jurisdiction
to ensure a fair trial by an
U.S. Water News, Sept. 2001
January 29-30, 2002
USEPA EDC Workshop
28
-------
What Are the Critical Research
Questions?
Effect relative to other Effects
Occurrence, Fate and Transport
Treatment Effectiveness
Suitable Operational Surrogates
January 29-30, 2002 USEPA EDC Workshop 29
-------
Sorption & Transport of Hormonally Active Agents,
Initial Laboratory Results
P. Suresh C. Rao & Linda S. Lee
Purdue University, West Lafayette, IN
Carl G. Enfield
NRMRL - USEPA, Cincinnati, OH
&EPA
EnwoimienlalPratectfin
Agency
-------
Project Team
Purdue University:
School of Civil Engineering
• Suresh Rao, Professor (Co-Pi)
• Ajit Sarmah, Post-doc
• Nathan Haws, PhD student
• Ryan Hultgren, PhD student
Purdue University:
Agronomy Department
• Linda Lee, Professor (Co-Pi)
• Sylvie Brouder, Associate Professor
• Maurilio Oliveira, Post-doc
• Steve Sassman, Lab Manager
• Troy Strock, MS student
US EPA- NRMRL:
• Carl Enfield
US EPA- NRMRL:
• Andy Avel, Project Officer
-------
Project Funding Sources
US EPA - NRMRL (2-yr project): (~ $100K for 2 yrs)
• Emphasis on EPA-designated Endocrine Disrupting Chemicals
• Primarily reproductive hormones
• National perspective
Purdue University (2-yr project): (~ $250K for 2 years)
• Emphasis on animal growth hormones
• Primarily confined animal feeding operations (swine)
• Indiana watersheds perspective
-------
DECEMBER 1 HOG INVENTORIES
SELECTED STATES, 1989-2000
20.0
15.0
10.0
5.0
0.0
1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000
Year
IL
IN
IA
MN
MO
ME
NC
U.S. HOGS BY SIZE GROUP
PERCENT OF OPERATIONS AND INVENTORY, 2000
Size Groups
Significance of CAFOs
in Indiana
Number of hogs in IN constant
since 1989 at« 5 million head
Small operations decreasing;
large operations increasing.
Steady decline in total number
of hog operations since 1979
from 25,000 to 4,500.
Over 60% have > 2000 hogs
Now large amount of manure
per area produced
Large amount of growth
hormones & antibiotics used
-------
Initial Project Objectives & Tasks
Identify major growth hormones used in swine CAFOs
and reproductive hormones
Conduct literature search for data on use, environmental
fate processes, & analytical methods
Conduct laboratory experiments to determine primary
fate & transport parameters (Koc, Kow, solubility, t1/2, etc.)
Conduct column experiments to determine sorption,
transformation and transport parameters under
steady flow conditions in saturated media
Monitor transport & fate of selected Pharmaceuticals
in tile-drained field plots receiving hog-manure application
Modify existing models to evaluate field data
-------
Hormonally Active Chemicals Selected
• Reproductive Hormones
•17(3-estradiol (natural)
•17oc-ethynyl estradiol (synthetic)
• testosterone
• Animal Growth Hormones
• carbadox
• tylosin
• tetracyclines (chlor- and oxy-)
• bacitracin
• bovine growth hormone (BGH)
-------
Soils & Sediments Selected
EPA Soils/Sediments %OC %clay pH
• EPA-1 (MN) 0.22 6 7.3
• EPA-14 (WVA) 0.48 64 4.3
• EPA-23 (IL) 2.38 69 7.1
Soils
• Bloomfield-3 (IN) 0.36 8 6.4
• Drummer-1 (IN) 2.91 21 7.2
• Toronto-4 (IN) 1.34 21 4.4
• Chalmers-6 (IN) 1.20 16 6.5
• Milford-21 (IN) 0.41 36 7.4
• Eustis (FL) 0.40 2 5.5
-------
Estrogens & Androgens
17(3-estradiol (272.4 g/mol) Testosterone (288.4
OH OH
Cht
Cht
0
17a-ethynyl estradiol (296.4 g/mol)
-------
en
.^
CO
O
16
12
8
4
0
17(3-estradiol
LogKoc = 3.38 + 0.18
0
1
EPA#1
A Drummer
50
40
17a-ethynyl estradiol
LogKoc = 3.03 + 0.1
• EPA#1
O Bloomfield
A Drummer
0
Cw (ug/mL)
120
100
^ 80
"o)
3, 60
CO
Cw (ug/mL)
Testosterone
Log Koc = 3.26 + 0.<
• Toronto
O Bloomfield
0
1234
Cw (ug/mL)
-------
Carbadox
0
• Drummer
D Bloomfield
A EPA#14
O EPA#1
O
Log Koc = 3.95 ± 0.20
8
10
Cw (ug/g)
• Used in swine pre-sbarter feeds as growth
pro m oter
• Aqueous solubility 51 mg/L
• Desoxy degradation product (in kidneys & liver) is
carcinogenic, m ore pdar & water soluble
-------
Carbadox on EPA-1 Soil Column: Concentration Effects
o
Non-equilibrium
F ~ 0.2
KT^O.005 fir7
1 ppm
5ppm
10 ppm
20
40
60 80 100 120
Pore Volumes
140 160
180
-------
Tylosin (A)
C46H77NO17 916.1 g/mol
Widely used in swine CAFOs
Weak base - pKa =7.1
Sw - 5,000 mg/L
pH 4-9 relatively stable
pH < 4, acid hydrolysis
pH > 7 Aldol forms
Kd: 2 to > 3000 L/kg
AtWQFS:upto Ippmi
field-applied manure
o
E
O
OH
1.5
1
0.5
0
• EPA 23 (pH 7.1; 69% clay)
D Drummer (pH 7.2; 21% clay)
A Chalmers (pH 6.5; 16% clay)
A Bloomfield (pH 6.4; 8% clay)
X Eustis (pH 5.5; 2% clay)
A A
0
0.002 0.004 0.006 0.008
Cw (mmol/L)
-------
Chlortetracycline Stability
in Na2HPO4 buffer
*pH8.2
•pH6.3
PH4.5
Time (minutes)
Sorption by Soils in 0.01 N CaCI2:
Single Point Kd values of 1000 to
1500 L/kg assuming sorption by
difference estimated for 6 soils
ranging in pH (2.5 - 6.4) and
%OC (0.2-3%).
50
Hydrolysis half-lives vs pH
t/> 30
(D
= 20
M—
x 10
0
4
5
8
pKa3 =96
+
HN(CHJ.
CONH,
pKa, =3-3
PKa2 = 78
-------
Nodal Drain
Preliminary Numerical Experiments
Using HYDRUS-2D
Homogeneous (Ksat = 0.26 cm/hr)
10m
ie?T*'*StW!P'-~- •
_.._ ^-flaifti. **-^sat
Jfe-!^-^-;^-
iBMWi^v^i^.
.
N °3 <0 A
— 0 200 -
0 500 -
H
L
°>
Hour
J
IF
JIT
Isotropic
Horizontal Anisotropy
Vertical Anisotropy
-------
HYDRUS-2D Simulation of Tracer Transport
Initial distribution of five
applied tracers
Distribution of five tracers
at t= 8000 hrs
-------
Conceptual JVJOCJB!: (Heterogeneouscase)
Vertical preferential pathways
only effective in near-tile area
Ks Scaling Factors:
Std Dev. 1
x-correl. 10cm
z-correl. 100cm
-------
Recovery of Multiple Tracers
4.50E-03
4.00E-03
— All
x 4.5-5.5
— 0-4.5,4.5-10
5.00E-04
O.OOE+00
500
1000
1500
2000
Hour
2500
3000
3500
4000
Note: Less than 0.1% of total applied tracers appear!
-------
Single Event Transport Simulations (con't)
Homogeneous
_o
"Si
SC-homo
SC-vhet
Loam vhet
SC vhet2
SC vhet2 trunc
Loam homo
O.OOE+00
Heterogeneous
500
1000
1500
2000
Hour
2500
3000
3500
4000
g
-------
Facilitated Transport of EDCs??
• Estrogens and testosterone have high sorption coefficients,
suggesting limited leaching potential in soils.
• Facilitated transport of EDCs bound to mobile colloids (soil or
manure) or complexed with DOC may be a major contributing
factor to transport to surface & groundwater.
• Need to evaluate factors that contribute to preferential flow
(e.g., macropores), and generation of DOC & mobile colloids.
• Desorption and bioavailability of bound EDCs??
• Field-applied manure at WQFS: 2% by weight of water-
soluble organic material in a single batch equilibration
(4g:20ml_) in preliminary estimates.
-------
Planned Activities
• Develop a comprehensive data set on environmental fate
& transport parameters for the selected HAAs and selected
soils/sediments (Purdue)
• Conduct lab column experiments (packed & undisturbed
soil columns) to evaluate leaching potential and
persistence under transient water flow conditions (Purdue)
• Conduct field experiments at the Purdue Water Quality
Field Station (PWQFS) to evaluate tile flow water quality in
plots receiving animal manure applications (Purdue & EPA)
-------
Planned Activities (Continued)
Develop a transport model to describe HAA transport data
from soil column studies (Purdue & EPA)
Develop a Lagrangian numerical simulator for describing
field data from the tile-drained plots at the Purdue WQFS
(Purdue & EPA)
Identify Indiana field sites at which surface water quality
can be monitored for HAAs (Purdue)
Collect water samples near CAFOs and from impacted streams
for HAA analyses (Purdue) and bioassays (EPA)
-------
Residential Indoor Air and Dust
Measurements of Phthalates and
Other EDCs
Ruthann Rudel, Silent Spring Institute
US EPA Effective Risk Management of
Endocrine Disrupting Chemicals
Cincinnati, Ohio -- January 29, 2002
Silent Spring Institute
-------
Searching for Preventable Causes of
Women's Diseases
Ra search ing the Envir
SF R I
INSTITUTE
Silent Spring Institute
-------
Collaborators and Funders
- Silent Spring Institute
» Ruthann Rudel and Julia G. Brody
- Harvard School of Public Health
» Jack Spengler and Jose Vallarino
- Southwest Research Institute
» Dave Camann, Paul Geno, Alice Yau,
Michelle Ortiz
Funding from Massachusetts Department of Public Health,
Boston Affiliate of the Susan G. Komen Foundation
Silent Spring Institute
-------
Why an exposure study?
Exposure assessment is weak link in:
- Environmental epidemiology
- Risk assessment, esp. mixtures, indoor air
- Endocrine disruptors research
See EHPDec. 2000, GAO reports, EPA EDSTAC,
NRC Report on Hormonally Active Agents,
NTP-CERHR Phthalate reports ...
Silent Spring Institute
-------
Why an exposure study?
Widespread concern about environmental pollutants
and breast cancer
EDCs and animal mammary carcinogens are promising
directions for study
Exposure data can help to
» ID chemicals and mixtures with common or high exposures as
priorities for research and regulatory policy
» ID highly exposed populations
» ID major sources of exposure
Silent Spring Institute
-------
To address these needs . . .
we developed and applied new methods to
detect a broad range of compounds identified
as hormonally active agents or animal
mammary carcinogens and applied them to
indoor air and dust samples
Silent Spring Institute
-------
Study Design (n=120 homes)
Number of Target Compounds
Chemical Group
Pesticides
Alkylphenols
Phthalates
Other estrogenic phenols
PCBs, PAHs
Parabens
Misc.
Estrogenic activity
(E-SCREEN bioassay)
Dust
Air
Urine Interview GIS
Silent Spring Institute
-------
Methods
Sample collection and analysis
- Dust from living area using teflon crevice tool, collected into
cellulose thimble, sieved to < 150 micron
- 24-hour air sample using high-vol pump, URG cartridge with
quartz filter, PUF, XAD2, particles < 5-10 micron, ~ 13 m3
- Two extraction/analytical methods (Rudel et al. 2001, JAWMA
51:499-513); phenols extracted with DCM, derivatized, GC/MS-
SIM; neutrals Soxhlet extracted in ether/hexane, GC/MS-SIM.
Subject selection
- Subset of women in case-control breast cancer study, over 65
years old, in current home > 10 years, oversampled high and low
self-reported pesticide users.
Silent Spring Institute
-------
10000.0
o
'•p
2
o
o
c
o
o
E
o
O
•5
c
.5
TJ
o
1000.0
100.0
10.0
1.0
0.1
Relative Abundance of Chemicals in Indoor Air Samples
Median of 30 Homes
oPPh2
NP1E
4tBPh
.PrPx
OP1 EO
AMePa
NP2EO ^^DCPh
^Hept
Diaz
PCB52 SgChk^r
• aChlor
- DEPt
- DBP
- DEHPt
- DIBPt
- BBPt
DEHA
_ DPPt
*APEO •Misc oPAH AParaben *PCB •Pesticide + Phenol -Phthalate
Silent Spring Institute
-------
Relative Abundance of Chemicals in Indoor Air Samples
90th Percentile of 30 Homes
1 UUUU.U
-—. -i nnn n
^^ IUUU.U
_c_
o
o
o
— i nn n
e of Che mica
* c
D C
D C
30th Percent!
->• c
D C
— ' 1 .U
O-i
. 1
- DEPt
m oPPh2
\ oPPhl . DBP
- DCPt+DEHI'
* NP .DEHPt
" DIBPt
_ BBPt
* NP1EO prpx + 4tBPh _ DEHA
^ OP1EO .. ChlPy
gChlor § ygf - DP"
Diaz 9 aChlor
^ NP2EO 4NPh ^ PCB52 • Carb
• OP2EO * 2'4DCPh • 3'5'6TCPy - DCP
•sr +—
^APEO aMisc OPAH AParaben *PCB •Pesticide + Phenol -Phthalate
Silent Spring Institute
-------
1,000.00
•5?
_3_
"E"
O
'•p
£
o
o
E
o
O
•5
c
w
TJ
O
100.00
10.00
1.00
0.10
0.01
Relative Abundance of Chemicals in Indoor Dust Samples
Median of 30 Homes
, NP
NP2EO
0 BaP
O BaA A MePa
tperm
A OP2EO
OP1EO *4NPh
Carb
DDT
BBPt
DINP
DBP
DOPt
DEHA
DHPt
+ BPA
1
PrPx
Folp + 4,4bVA
gChlor
aChlor
Misc o PAH AParaben ^PCB •Pesticide + Phenol -Phthalate
Silent Spring Institute
-------
Relative Abundance of Chemicals in Indoor Dust Samples
90th Percentile of 30 Homes
1 UUU.UU
4 nn nn
Q) IUU.UU
l>
o
o
0
W -in nn
ile of Chemic
->• c
D C
D C
90th Percent
D
•>• C
D C
U. 1 U
n m
- DCPt+DEH
- DEHPt
- DINP
- BBPt
_ tPerm _ DOPt
_ cPerm _ DEHA
+ NP2EO : PjPBO - °HPt
+ NP1 EO
NP ^NP1EC °BaP Carb JJ MY - DEPt
m-vr " DIBPt
QBaA • °DJ _i_ BPA
k IVtePs A PrPx
A PCPh «,. ._ nr^Dt
r^rM_
-------
Pesticides detected in at least
50% of homes.
o-phenylphenol
permethrin
piperonyl butoxide
carbaryl
methoxychlor
heptachlor
4,4'-DDT
propoxur
pentachlorophenol
chlorpyrifos
chlordane
folpet
diazinon
10% of homes:
All those plus
bendiocarb
3,5,6-trichloro-2-pyridinol
dieldrin
dicofol
chlorothalonil
endrin
4,4'-DDD
4,4'-DDE
Silent Spring Institute
-------
Classes of chemicals detected, in order of
relative abundance
phthalates (esp. DEP, DBF)
0-phenyl phenol
nonylphenol, APEOs
pesticides
4-t-butyl phenol
methyl paraben
PCB52
Dust
- phthalates (esp. DEHP)
- nonylphenol, APEOs
- pesticides
- PAHs
- bisphenol A
- parabens
- PCBs
Silent Spring Institute
-------
Comparison with EPA Region 9 PRGs
Compounds exceeding
residential soil PRGs
- Benz(a)pyrene
- Benz(a)anthracene
- PCB 52, 105, 153
- Heptachlor
- Chlordane
- Pentachlorophenol
- Dicofol
- Dieldrin
- DDT
- Bis(2-ethylhexyl)phthalate
Compounds exceeding
air PRGs
- Chlordane
- Heptachlor
- PCB 52
Silent Spring Institute
-------
Exposure Levels-DBP, DEHP
Dibutyl Phthalate
Di(2-ethylhexyl) phthalate
120
100
ro 80
2 60 -
40
20 -
Ma
x i
*
95%
E PA-IRIS
k X 1953 rat
mortality
Foster
2000 10-
day
prenatal-
repro tract
develop.
Additional
r 1 0-fold
• X needed ?
med I ^
CDC
Urine
Rudel
Air Dust
Reference
Dose (RED)
50
45
40
> 35 -
ro
o> 30
§ 25 ^
15
10
5 -
CDC
Urine
Rudel
Air Dust
Ma
X A
1
95%
med
k
E PA-IRIS
. * 1953 rat
]iver
wght
CERHR 2000
i r - but new
i T
* study needed
1 A
Reference
Dose (RED)
Silent Spring Institute
-------
Relative Abundance-Phthalates
400
200
Dust (mg/g)
600
400
200
Air (ng/m3)
180
160
140
120
100
DDEHP
• DINP
DBBP
• DBF
DDEP
OOP
DDCHP
Urine (ng/ml)
From CDC National Report Card, 2001
Silent Spring Institute
-------
Implications
A large number of lexicologically important
chemicals (EDCs/MCs) are widespread in
indoor environments
- Exposure data helps prioritize chemicals and
mixtures for testing; identify major sources of
exposure
- Need to understand health effects in order to
limit exposure to the most important agents
Silent Spring Institute
-------
www.SilentSpring.org
Silent Spring Institute
-------
Welcome
Gregory Sayles, NRMRL
EDC Workshop January 29, 2002
Welcome to the CD-ROM version of the workshop, Effective Risk Management of Endocrine
Disrupting Chemicals. This workshop was held in January 2002 in Cincinnati and sponsored by EPA's
Office of Research and Development and EPA's National Risk Management Research Laboratory.
Hello, my name is Greg Sayles. I lead the risk management portion of EPA's endocrine disrupting
chemicals research program.
To date, most research on endocrine disrupting chemicals (EDCs for short) has focused on health and
ecosystem effects. With this workshop, our goal is to begin the discussion of risk management of
suspected EDCs. To the best of my knowledge, this workshop is the first technical meeting ever
conducted that focuses on risk management of suspected EDCs.
This workshop has been designed for stakeholders of environmental EDC problems, including:
• researchers from government, university and industrial labs who will develop risk management
approaches
• Federal, State and local regulators who are charged with making risk management decisions
• engineering professionals who will implement risk management approaches
• industries associated with suspected EDCs who need to know the current state of the art in risk
management of EDCs
Sooner or later many of you and your colleagues may be asked for input on managing the risk of
EDCs. This workshop introduces you to the tools you will need:
• we begin with introductory talks about risk management - first in general then, how our colleagues
in Europe are approaching risk management of EDCs
• next, we present speakers to bring you up to date on health effects and exposure assessment for
suspected EDCs
• finally, we show the current status of applied risk management approaches for suspected EDCs:
Many people were involved in the planning and presentation of this workshop and in the development
of this CD-ROM. I greatly appreciate all their hard work. Please see the credits page for more
information.
We hope you find this multimedia, interactive CD-ROM a useful introduction to the current state of risk
management of endocrine disrupting chemicals. We welcome your comments.
Thank you and enjoy.
-------
EPA's
RISK MANAGEMENT
EVALUATION OF EDCs
Gregory D. Sayles
U.S EPA Office of Research and Development
National Risk Management Research Laboratory
Cincinnati, Ohio
"errvimninentaf solutions today for a safer
-------
Presentation Overview
RM
Research
Questions
-------
What is a Risk Management
Evaluation (RME)?
The current understanding of risk
management of an
environmental challenge
Important sources of
problem,
environmental sinks
Currently
available RM
approaches
Identifies
knowledge
gaps, research
needs
-------
Uses for RMEs?
<*Defines the environmental problem
*Gives current and future RM options to
stakeholders (regulators, public interest
groups, etc.)
<*Allows environmental consultants /
engineers to asses current skills and
future investment
* Assists in research planning
-------
Development of the RME
Concept
*NRMRL developed several pilot RMEs
for internal review
*One pilot RME was for EDCs
<*Protocol for building RMEs drafted by
NRMRL
-------
The RME for Endocrine
Disrupting Chemicals
Writing Team
Gregory Sayles, lead writer
John Cicmanec
Steve Hutchins
Paul Lemieux
Carl Potter
Kathleen Schenck
-------
Summary of Content
For a list of likely EDCs, describes
* Known health and eco effects
* Significant sources to the environment
* Significant exposures / environmental sinks
* Risk management tools
established
needed
-------
RME Structure
* Chapter 1
* Chapter 2
* Chapter N-1
* Chapter N
-------
RME Structure
* Chapter 1
* Chapter 2
Intro to EDC problem
Regulatory mandate
Goals of document
EDCs considered
* Chapter N-1
* Chapter N
-------
RME Structure
* Chapter 1
* Chapter 2
* Chapter N-1
* Chapter N
Known health, eco effects
Known sources
Known reservoirs
Useful RM approaches
- established or new
-------
RME Structure
* Chapter 1
* Chapter 2
* Chapter N-1
* Chapter N
Version 1.0
• Alkylphenols
• Steroid hormones: biogenic
and pharmaceutical
• PCBs
• Chlorinated dioxins and
furans
• DDT and DDE
-------
RME Structure
* Chapter 1
* Chapter 2
* Chapter N-1
* Chapter N
Later versions
• Bisphenol A
• Phthalates
• Atrazine
• Methoxychlor
• Endosulfan
• Others?
-------
RME Structure
* Chapter 1
* Chapter 2
* Chapter N-1
* Chapter N
Summary of
• RM tools available
• RM research needs
-------
Alkylphenolics, especially
Nonylphenolics
*EDCs of interest:
Nonylphenol
Nonylphenol ethoxylates
Nonylphenol ethoxy carboxylic acids
*Eco effects - lab and field observation
of estrogenic activity
'MHuman effects - none, except in vitro
tests using human cell lines (estrogenic)
-------
Parent Chemical
Nonylphenol polyethoxylate
OH
CT n
NPnEO
Not estrogenic
-------
Generation
of EDCs
More
hydro-
phobic
NP1EO
estrogenic
NP
-------
Uses for Nonylphenolics
*Nonylphenol
Antioxidant, lube oils
*Nonylphenol ethoxylate surfactants
Industrial and domestic detergents
Plastics, ag chemicals, paper production
Not EDCs
Biotransformed into EDCs in
• sewage treatment plants
• the environment
-------
Market for NPE
350 - 500 million Ibs / yr sold
NPE used in: Home
detergents
Industrial
products
Plastics
Textiles
Paper
Ag chemicals
Laundry detergent
Hard surface cleaner
Industrial
cleaners
Metal, vehicle,
hard surface cleaning
Commercial laundry
-------
Likely Sources
*Wastewater treatment plants •
Domestic and industrial
*Pulp and paper mills •
* Pesticide use •
^Textile mills probably
-------
Source Data
Sewage Treatment Effluent - Nonylphenol
1000
NP
100
10
-------
Reservoir Data
Nonylphenol:
* Sediments - Canada < 44 mg/kg
Surface Water - UK < 1 9
Groundwater - Cape Cod 30
* Drinking water- Spain < 0.14
*Air - NY City area < 25 ng/m3
-------
Risk Management Approaches
* Phase out / Product substitution
e.g., EU and alcohol ethoxylates
* Alter sewage treatment plant operation
Improve aqueous effluent
• Add PAC to secondary treatment
• Add tertiary GAG treatment
• Add tertiary granular medium filtration
• Alter secondary treatment process variables
-------
Research Questions
*What is the performance of unit
processes in STPs?
*Do current sludge disposal methods
perform well?
*What is the capacity of aquatic sediments
to manage the input of APs?
-------
Research Questions
*Are other sewage treatment processes
significant sources?
Septic systems
Constructed wetlands
*Are conventional drinking water treatment
methods adequate? If not, are advanced
treatment technologies?
-------
Biogenic / Pharmaceutical
Steroid Hormones
*EDCs of interest:
natural steroid hormones, especially
estrogens
Veterinary steroid hormones
<*Effects - humans/mammals, birds, fish
have steroid hormone receptors
-------
Structures of Some Hormones
estradiol
OH
HO
O
O
TbA
OH
OH
HO
O
TbOH
ethinyl estradiol
-------
Sources
* Sewage treatment
plants
* Concentrated animal
feeding operations
(CAFOs)
-------
Sinks / Reservoirs
<*Surface water
Widely observed 1-10 ng/l
^Sediments
Expected accumulation - log Kow = 3-4
<*Ground water
Estradiol < 60 ng/l (CAFO source)
-------
RM Approaches
*Hormones aerobically biodegradable
*Will partition to activated carbon
-------
RM Research Questions
* What is the fate of steroid hormones in unit
processes of STPs?
*How can STPs be improved to increase
treatment?
*Are sludges significant reservoirs for steroids?
*Are sediments a significant sink? Are natural
processes protective?
-------
RM Research Questions
* What is the hormone content of animal waste?
*Do current waste management practices at
CAFOs minimize environmental exposure? If
not, how can they be improved?
*Does conventional drinking water treatment
remove hormones? If not, are innovative
approaches available?
-------
PCBs
* Research questions:
What is the short term risk
associated with dredging
sediments?
What is the long-term ^^m
stability/effectiveness of capping
sediments?
When is natural
attenuation/recovery of sediments
effective?
-------
Chlorinated Dioxins and Furans
Cl\ ^^ ^CL ^^ ^Cl
Research Questions
How can incinerators be operated to
produce less chlorinated dioxins and
furans
What combustion processes produce
more/less endocrine active compounds?
How can the high producing processes be
modified to minimize release?
-------
RME - Evolution
*The RME is a living document
Version 1.0 will be completed Fall '02
Later versions roughly bi-annually
Peer reviewed
-------
"ewrfcamnealal salatians tads? for a safer foiaomcm"
>s EDC Research Program
* Project areas based on RME research
questions
*EPA Principle Investigator driven
-------
's EDC Research Areas
Sewage Treatment
EDC fate within STPs
• Paul McCauley / Dick Brenner
EDCs in sediments
• Greg Sayles / Marc Mills
Drinking water treatment
Effectiveness of
conventional and advanced
treatments
Kathy Schenck
-------
EDC Research Areas
"etrriFcasnaifal solutions today for a safer tomorrow*
Confined Animal Feeding
Operations
CAFOs as a source of EDCs
Effectiveness of waste
management practices
Steve Hutchins / Carl Enfield
Pollution Prevention
Tool development for
identification of EDC substitutes
Doug Young
-------
EDC Research Areas
a safer tomorrow*
* Combustion Characterization
EDC content of various process
effluents
Brian Gullett
*Bioassays for RM
Performance Evaluation
Bioassays show big picture
performance of RM
Carolyn Acheson
-------
s EDC Research Areas
"eaviromiKiaalsolutions today for a safer toatomon'"
^Technical Information Transfer
Summary documents, web site, workshops
-------
RM Decision-Making Tools
Risk management decisions for EDCs will be
required more frequently
What RM tools do you have?
This Workshop (in person, CD-ROM)
Risk Management Evaluation of EDCs
EPA's RM of EDCs web page
www.epa.gov/NRMRL/EDC
What RM tools are being developed?
EPA's RM research program
Others' research
-------
Summary
*Risk Management Evaluation ofEDCs
Will help you follow state of the art in RM
Provide input to research agendas
*RME drives NRMRL research
*Build your tool box
-------
Biological Fate of Estrogenic
Compounds Associated with Sewage
Treatment: A Review
Gregory Sayles
U.S. EPA
Tamara Marsh
Elmhurst College, IL
Elmhurst College
-------
Purpose
Give you current
thinking on fate of
EDCs in sewage
treatment systems
- Available data
- Qualitative discussion
•
-------
Most Prevalent Estrogenic
Chemicals found in STP Effluent
Alkylphenolic compounds
- Biodegradation products from APE surfactants
• Nonvlphenolics
• Octylphenolics
Steroid hormones
- Natural (estradiol, estrone, estriol)
- Synthetic (ethinyl estradiol)
-------
Typical Large STP
influent
Bar
Screen
Grit Primary
Chamber Clarifier
Aeration
n
Secondary
Clarifier
effluent
Dewatered
Sludge
Chlorination
Tank
De-
watering
Anaerobic
Digestion
Thickener
Sludge Treatment / Disposal
Water
-------
STPs Designed for...
BOD removal
Suspended solids removal
pH neutralization
N, P removal
Pathogen removal
-------
Current Knowledge Limited
For nonylphenolics:
• Parent NPE are a distribution of EO units
• > 20 isomers of the nonyl group
• Analytical standards for NPE metabolites difficult to
obtain
For steroids and nonylphenolics:
• Analytical methods not standardized
• Low detection limits needed (ng/l)
Few biodegradation studies published
-------
NPEs - STP Influent / Raw Sewage
~ 500 million pounds used per year in U.S.
Institutional: laundry detergents, janitorial and vehicle
cleaners
Household: cleaners and personal care products
Industrial: plastics, textiles and pulp and paper
processing, ag chemicals
Total nonylphenolics 500 - 2500 ug/l
Total estrogenic products 1-50 ug/l
-------
Biological Transformation of APEs
Aerobic
O'Jn
OH
AaPE
Parent surfactant
"nonylphenol polyethoxylate"
-------
Biological Transformation of APEs
Aerobic
A9PE2
A9PEt
-------
Biological Transformation of APEs
Aerobic
AoPE.C
A9PEtC
-------
Biological Transformation of APEs
Aerobic
-------
Biological Transformation of APEs
Aerobic
-------
Biological Transformation of APEs
Anaerobic
-------
Nonylphenolic STP Balance
Rome, Italy (Di Corcia, 2000)
Cone., ug/l 40
-------
NPE Metabolites Partition to Sludge
EDC
log (Kow)
NP
NP1EO
NP2EO
CNP1EC
4.5
4.2
4.2
«4
-------
Likely Fate in STP
Aeration
Tank
Secondary
Clarifier
A9P
AoP
Anaerobic
Digestion
-------
Effluent Fate
> Expect alkyphenolics in
> Surface water (up to 20 ug/l)
> Sludges (up to 50 mg/kg)
> Sediments (up to 4000 mg/kg)
-------
Influent Estrogens
estradiol
OH
estrone
HO
estriol
ethinyl estradiol
-------
Amount of Estrogens Excreted
1000
ug/day 100
10
Pregnant
Menstruating
Menopausal
Male
•Only for those taking ethinyl
(=0 for males, pregnant)
-------
STP Influent - Raw Sewage
* Estradiol
* Estriol, estrone
•:• Ethinyl estradiol
ND- 50ng/l
ND-100ng/l
ND- 10ng/l
-------
Estrogens in Activated Sludge
From 6 STPs in Tennessee (Layton, 2000)
o
'"B
CD
CD
O
CD
100
80 -
60 -
40 -
20 -
0
ethinyl estradiol
0
10 15
Hours
20
25
-------
Example STP Removal - Rome Italy
Note log scale
100^
ng/l
influent
effluent
estradiol estrone estriol
ethinyl
See Baronti, 2000
-------
I Biodegradation of Estrogens
Aerobic
• All biodegradable, ethinyl slowest
• Nitrification may increase rate
Anaerobic
• Little known
• Little biodegradation expected
-------
Partition to Sludge, Sediments?
EDC
Ice
Ethinyl estradiol
Estradiol
Estrone
Estriol
4.2
3.9
3.4
2.8
No sludge, sediments field data to date
-------
Overall Impact of Various EDCs in
STPS (Johnson, 2001)
Chemical
Estradiol Equiv.
(in vivo studies)
Level of Concern
Ethinyl estradiol
Nonyl-, octyl-phenol
Estrone
Estradiol
Estriol
APEs, APECs, CAPEs
12
2-20
2.5
1.0
0.02
High
Moderate
Moderate
Moderate
Lowest
Low?
-------
Summary
Current limited data
- Aerobic biodegradation produces estrogenic alkylphenolics in
STPs
- No evidence of destruction of alkylphenolics in STPs
- Steroid hormones are aerobically biodegradable
• Retention time in STP appears to be short
- Sludge will accumulate low MW alkylphenolics and hormones
Need detailed studies of EDC fate within the STP
Are other sewage treatment systems removing EDCs?
- Septic systems?
- Small community treatment, constructed wetlands?
-------
Arcand-Hoy, L. D., A. C. Nimrod and W. H. Benson (1998). "Endocrine-Modulating
Substances In The Environment: Estrogenic Effects of Pharmaceutical Products."
International Journal of Toxicology 17: 139-158.
Baronti, C., R. Curini, G. D'Ascenzo, A. Di Corcia, A. Gentiliand R. Samperi (2000).
"Monitoring Natural and Synthetic Estrogens at Activated Sludge Sewage Treatment
Plants and in a Receiving River Water." Environmental Science & Technology 34(24):
5059-5065.
Belfroid, A. C., A. Van der Horst, A. D. Vethaak, A. J. Schafer, G. B. J. Rijs, J. Wegener
and W. P. Cofino (1999). "Analysis and Occurence of Estrogenic Hormones and Their
Glucuronides in Surface Water and Waste Water in The Netherlands." The Science of
the Total Environment 225: 101-108.
Benne, D. T. (1999). "Review of the Environmental Occurence of Alkylphenols and
Alkylphenol Ethoxylates." Water Quality Research Journal of Canada 34(1): 79-122.
Bolz, U., H. Hagenmaier and W. Korner(2001). "Phenolic Xenoestrogens in Surface
Water, Sediments, and Sewage Sludge from Baden-Wurttemberg, South-West
Germany." Environmental Pollution 115: 291-301.
Desbrow, C., E. J. Routledge, G. C. Brighty, J. P. Sumpter and M. Waldock (1998).
"Identification of Estrogenic Chemicals in STW Effluent. 1. Chemical Fractionation and
in Vitro Biological Screening." Environmental Science & Technology 32(11): 1549-1558.
Di Corcia, A., R. Cavallo, C. Crescenzi and M. Nazzari (2000). "Occurrence and
Abundance of Dicarboxylated Metabolites of Nonylphenol Polyethoxylate Surfactants in
Treated Sewages." Environmental Science & Technology 34(18).
Fawell, J. K, D. Sheahan, H. A. James, M. Hurst and S. Scott (2001). "Oestrogens and
Oestrogenic Activity in Raw and Treated Water in Severn Trent Water." Water Research
35(5): 1240-1244.
Ferguson, P. L., C. R. Iden and B. J. Brownawell (2001). "Distribution and Fate of
Neutral Alkylphenol Ethoxylate Metabolites in a Sewage-Impacted Urban Estuary."
Environmental Science Technology 35(12): 2428-2435.
-------
Furbacker, M., A. Breithofer and A. Junghauer(1999). "17beta-Estradiol: Behavior
During Waste Water Analyses." Chemosphere 39(11): 1903-1909.
Giger, W, M. Ahel, M. Koch, H. U. Laubscher, C. Schaffnerand J. Schneider (1987).
"Behaviour of Alkylphenol Polyethoxylate Surfactants and of Nitrilotriacetate in Sewage
Treatment." Water Science and Technology 19: 449-460.
Giger, W, P. H. Brunner and C. Schaffner (1984). "Nonylphenol in Sewage Sludge:
Accumulation of Toxic Metabolites from Nonionic Surfactants." Science 225: 623-625.
Hale, R. C., C. L. Smith, P. O. de Fur, E. Harvey, E. O. Bush, M. J. La Guardia and G.
C. Vadas (2000). "Nonylphenols in Sediments and Effluents Associated with Diverse
Wastewater Outfalls." Environmental Toxicology and Chemistry 19(4): 946-952.
Hesselsoe, M., D. Jensen, K. Skals, T. Olesen, P. Moldrup, P. Roslev, G. K. Mortensen
and K. Henriksen "Degradation of 4-Nonylphenol in Homogeneous and
Nonhomogeneous Mixtures of Soil and Sewage Sludge." Environmental Science &
Technology.
Johnson, A. C., A. Belfroid and A. Di Corcia (2000). "Estimating Steroid Oestrogen
Inputs Into Activated Sludge Treatment Works and Observations on Their Removal
From the Effluent." The Science of the Total Environment 256: 163-173.
Johnson, A. C., R. J. Wlliams and T. Ulahannan (1999). "Comment on "Identification of
Estrogenic Chemicals in STW Effluent..1. Chemical Fractionation and in Vitro Biological
Screening"." Environmental Science Technology 33(2): 369-370.
Korner, W., U. Bolz, W. Susmuth, G. Hiller, W. Schuller, V. Hanf and H. Hagenmaier
(2000). "Input/Output Balance of Estrogenic Acitve Compounds in a Major Municpal
Sewage Plantin Germany." Chemosphere 40: 1131-1142.
La Guardia, M. J., R. C. Hale, E. Harvey and T. M. Mainor(2001). "Alkylphenol
Ethoxylate Degradation Products in Land-Applied Sewage Sludge (Biosolids)."
Environmental Science & Technology 35(24): 4798-4804.
MaGuire, R. J. (1999). "Review of the Persistence of Nonylphenol and Nonylphenol
Ethoxylates in Aquatic Environments." Water Quality Journal of Canada 34(1): 37-78.
Marcomini, A., F. Cecchi and A. Sfriso (1991). "Analytical Extraction and Environmental
Removal of Alkylbenzene Sulphonates, Nonylphenol and Nonylphenol Monoethoxlate
from Dated Sludge-Only Landfills." Environmental Technology 12: 1047-1084.
-------
Routledge, E. J., D. Sheahan, C. Desbrow, G. C. Brighty, M. Waldock and J. P.
Sumpter (1998). "Identification of Estrogenic Chemicals in STW Effluent. 2. In Vivo
Responses in Trout and Roach." Environmental Science & Technology 32(11): 1559-
1565.
Sekela, M., R. Brewer, G. Moyle and T. Tuominen (1999). "Occurence of An
Environmental Estrogen (4-Nonylphenol) in Sewage Treatment Plant Effluent and the
Aquatic Receiving Environment." V\fater Science and Technology 39(10-11): 217-220.
Shishida, K., S. Echigo, K. Kosaka, M. Tabaski, T. Matsuda, H. Takigami, H. Yamada,
Y. Shimizu and S. Matsui (2000). "Evaluation of Advanced Sewage Treatment
Processes for Reuse of V\fostewater Using Bioassays." Environmental Technology 21:
553-560.
Ternes, T. A., P. Kreckel and J. Mueller (1999). "Behavior and Occurrence of Estrogens
in Municipal Sewage Treatment Plants - II. Aerobic Batch Experiments with Activiated
Sludge." The Science of the Total Environment 225: 91-99.
Ternes, T. A., M. Stumpf, J. Mueller, K. Haberer, R.-D. Wilken and M. Servos (1999).
"Behavior and Occurrence of Estrogens in Municipal Sewage Treatment Plants -1.
Investigations in Germany, Canada and Brazil." The Science of the Total Environment
m 225:81-90.
O Williams, R. J., M. D. Jurgens and A. C. Johnson (1999). "Initial Predictions of the
™ Concentrations and Distribution of 17beta-Oestradiol, Oestrone and Ethinyl Oestradiol
in 3 English Rivers." Water Research 33(No. 7): 1663-1671.
-------
Evaluation of Drinking Water
Treatment Technologies
for Removal of Endocrine
Disrupting Compounds
Kathleen Schenck and Thomas Speth, U.S. EPA, NRMRL
Laura Rosenblum, Steve Wendelken, Barry Pepich,
and Radha Krishnan, IT Corporation
Kent Mitchell and David Warshawsky, University of
Cincinnati
-------
Many of the chemicals identified as
potential endocrine disrupting compounds
(EDCs) may be present in surface or
ground waters used as drinking water
sources due to their introduction from:
* Domestic and industrial sewage
treatment systems.
* Wet-weather runoff.
-------
Basic strategies to decrease the potential risk
of adverse health effects associated with the
presence of EDCs in drinking water:
* Protect drinking water sources from
contamination by EDCs.
* Remove EDCs, that may be present in
source waters, during drinking water
treatment.
-------
Compourrds to be evaluated
Estradiol
Estrio.
Ethynylestradiol
-------
Compounds to be evaluated
Progesterone
c=o
Testosterone
Dihydrotestosterone
-------
Additional compounds to be
evaluated in the future
* 4-nonylphenol (NP)
* 4-nonylphenol mono-ethoxylate (NP1E
* 4-nonylphenol diethoxylate (NP2EO) ^
* 4-nonylphenoxy carboxylic acid (NP1EC)
* 4-nonylphenoxy ethoxy carboxylic acid
(NP2EC)
-------
Technical approach
* Develop analytical methods to identify
and quantify the target compound
The approach will include concentrati
by solid-phase extraction, followed by
LC/MS.
-------
Analytical method for steroid
compounds
\.
Solid phase extraction: N^
* Baker C18 XF speed disks eluted with
methanol
Quantitation:
* Waters ZQ LC/MS, electrospray
* Xterra C18 column
* Single step gradient, 50 - 65% methanol in
ammonium hydroxide in water
* Single ion mode
-------
Single ion chromatograms of reagent water fortified at 1 ng/L
100% .
>
surrogate
bisphenol-c/fft
estriol^
ethynylestradiol
testosterone
progesterone
estradiol
d ihyd rote stoste rone
Time, mm
10.0
EI e c tro s p r a y
20.0 30.0
1 Electro spray +
-------
Technical approach (cont.)
* Evaluate the use of a reporter gene
assay, the MVLN assay, to detect the
presence/ removal of estrogenic activity
This assay uses a human breast cell
line (MCF-7) which has been stably 1
transfected with the firefly luciferase
gene.
-------
Technical approach (cont.)
•^. --
Conduct bench-scale evaluations of various
drinking water treatment technologies
including conventional treatment, granula.
activated carbon, softening and nanofiltratio,
* Pilot-scale evaluations may be conducted on
the treatment technologies that appear T
promising at bench-scale.
-------
This study will provide information on:
currently available drinking water treatment
technologies that can remove EDCs,
specifically the steroid hormones and the
nonylphenolic compounds.
* approaches to optimize these treatment
technologies for EDC removal.
* the need for additional management tools to
be developed for the removal of EDCs during
drinking water treatment.
-------
***
European Commission - DG ENV
Effective Risk Management of Endocrine Disrupting
Chemicals, 29-30 January 2002, Cincinnati, Ohio Ur
'European Community Strategy for
docnne Disruvters: Imvlementation
Kathryn Tierney,
Author: KT January 2002
-------
***
Contents of presentatio
European Commission - DG ENV
European Community (EC) Strategy for
Endocrine Disrupters COM(1999)706
Implementation to date COM(2001)262
+ Identification of substances
R&D
+ Legislative Action
Conclusions
Author: KT January 2002
-------
***
***
1. EC Strategy (1
European Commission - DG ENV
Need for further research
Need for in
Need for communication to the publi
eed for appropriate policy action
Author: KT January 2002
-------
***
***
1. EC Strategy (2)
European Commission - DG ENV
Short-term actio
o Establishment of priority list of substances for further
evaluation of their role in endocrine disruption
o Use of existing legislative instruments
o Establishment of monitoring programmes
o Identification of specific cases of consumer use
o Information exchange/international cooperation
o Communication to the public
o Consultation with stakeholders
Author: KT January 2002
-------
***
European Commission - DG ENV
Medium-term acti
ntification and assessment of EDs
oR&D
o Identification of substitutes
Long-term action
o Overall chemicals policy
o Water Framework Directive
o Other legislative actions/proposals
Author: KT January 2002
-------
***
***
2, Implementation to date (
European Commission - DG ENV
Identification of substance
Step 1 : Study Report by BKH, June 20
Itation of scientists. Member
KfETt
ority list of action
Author: KT January 2002
-------
***
***
ementation to date
European Commission - DG ENV
Identification of substances
BKH Report a reasonable starting point
Is D!US svnth
/natural hormones present in the envi
Al 553 candidate substances retained for further
et up iterative mechanism for updating
No duplication of work with ongoing risk
issessments under existing Community legislation
Author: KT January 2002
-------
***
***
ementation to date
European Commission - DG ENV
Identification of substances
Of 118 substances with evidence of endocrine
disruption or potential ED in BKH Report, 109 ar
already subject to regulatory measures:
o 16 obsolete plant protection products
o 14 banned in EU
o 22 with marketing restrictions in EU
oil subject to emission controls
o 46 currently on priority lists for risk assessment
Author: KT January 2002
-------
***
***
2, Implementation to date (4
European Commission - DG ENV
Identification of substances
Study to conduct in-depth evaluation ofl*
substances not currently addressed (12-18 months)
Study to gather information on 435 substances with
sufficient data in BKH Report (12-18 months)
Invite Member States (MS) to take ED into
during RA of 46 subs tan ~
ca
Author: KT January 2002
-------
***
***
2, Implementation to date (5
European Commission - DG ENV
R&D
4th Community Framework Programme for
R&D (1994-1998) » circa 8 M €/7 M $
5th Community FP for R&D (1999-2002)
ca 10.45 M 6/9.2
circa 20 M 6/17.6
to be awarded in 200
Author: KT January 2002
-------
***
***
ementation to date (6)
European Commission - DG ENV
\slative action
Overall chemicals policr
aper, 13 February 2001
horisation procedure for CMR/POPs
o Highlights need for further research on EDs —
on test methods, low dose effects, QSARs
o Rigorous testing for long-term effects for >100t
Author: KT January 2002
-------
***
***
ementation to date
European Commission - DG ENV
\slative action
Overall chemicals Dolicv cont'd
001
e su
uthorisation
when agreed scientifically valid test methods
and criteria are established
Author: KT January 2002
-------
***
***
ementation to date (8)
European Commission - DG ENV
Legislative acti
Water Framework Directive
• Directive 2000/60/EC and Decisio
2455/2001/EC establishing the list of priority
substances in the field of Water Policy
o 33 priority substances, 11 are candidate EDs
o Measures to be proposed within 2 years aimed
at ending or phasing out emissions, discharges
and losses within 20 years.
Author: KT January 2002
-------
***
***
ementation to date (9)
European Commission - DG ENV
Legislative act
General Product Safety Directiv
Key risk management instrument in short-
Recent revision allows simplification of
procedures and conditions for urgent
measures at Commumtv level
Author: KT January 2002
-------
***
***
1999
3. Conclus o
European Commission - DG ENV
Preparation of Communit
trategy
2000-2001 Preparatory activiti
identification of substances,
planning phases for R&D call for proposals
carrying out of technical studies
2001-2002 Implementation
launch of research projects
revision and preparation of legislative proposals
launch of new activities e.g. monitoring
Author: KT January 2002
-------
***
***
European Commission - DG ENV
Website address
http://www.europa.eu.int/comm/environme
ocum/99706sm.htm
Author: KT January 2002
-------
Endocrine/Estrogen Letter
Online Enterprises • 245 Visitacion Ave, Brisbane, CA 94005
Phone: (415) 467-8779 * Fax 415-468-3144* e-mail: info@eeletter.com
Vol. 8, No. 2 (163)
EC Risk Management. 2
EDO Wildlife Effects 6
EPA's EDO Strategy. 8
Exposure Assessment....9
Risk Man Approaches 11
Drinking water. 12
CAFOs. 14
Wastewater treatment.....16
Combustion sources 17
Cheap PCB Cleanup 18
EDC Alternatives 18
Endocrine/Estrogen Letter is
published 12 times a year.
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February 2002
E/E Conference Report
This special issue is dedicated to coverage of The Effective Risk
Management of Endocrine Disrupting Chemical (EDC) workshop held
in Cincinnati from January 29-30. The conference explored a number
of issues including the context of risk management of EDCs, effects of
EDCs on humans and wildlife, exposure assessment of EDCs, drinking
water treatment, concentrated animal feed operations, waste water
treatment, and the search for EDCs in combustion sources.
Greg Sayles, with the US EPA National Risk Management
Research Laboratory (NRMRL) said this was the first technical
meeting to focus on the risk management of endocrine disrupting
chemicals.
There was some speculation at the conference that moving to a
discussion on risk management is a bit premature, considering that the
work on confirming the endocrine disrupting properties of chemicals is
still in its infancy. But developing risk management strategies now may
make it easier to deal with any confirmed endocrine disrupters once
they have been conclusively identified.
Lee Mulkey with the NRMRL said there were three main things
that were different about the topic and how they went about organizing
the work.
First, EDCs have been identified as an emerging risk. Mulkey
explained, "This is an important idea because the EPA typically
responds and reacts to issues that come up, whether it is rivers that
burn or the superfund being spun out of Love Canal. This is an issue
where the EPA has had the foresight to consider what may or may not
be an initiative that requires new policy development.
Second, we need to worry about the footprints of EDCs. There
has not been much thought on EDCs as wastes, or how the environ-
ment can assimilate them.
Third, the idea of emerging risk is an integrative science. The idea
is to develop knowledge about risk management while we are still
learning about the effects and extent of exposure of EDCs. We can
figure out ways in which intervention might be appropriate and the
costs and benefits of various approaches.
Dr. Hugh McKinnon, Associate Director for Health, at the
NRMRL talked about the history of risk management within the EPA.
In 1983, the National Research Council put together the "Red Book"
which spells out the risk assessment/risk management paradigm. It
talks about the process of risk assessment, risk management, and risk
(Continued on page 2)
-------
Endocrine/Estrogen Letter
February 2002
E/E Conference Report
communication.
Risk assessment classically has consisted of
four steps: exposure assessment, hazard identifica-
tion, dose response assessment, and risk character-
ization. The primary problem in risk assessment has
been dealing with uncertainty.
McKinnon said where legal or political man-
dates are clear, or where we have unambiguous
present risk, we will tolerate a great deal of uncer-
tainty in risk assessment and remediation, especially
if the costs are not high. It often turns out that as we
develop a risk assessment, we get more data. We
are able to reduce that uncertainty to a point where
we can make a decision and hopefully that results in
a lowering of risk. In an ideal case, if we have similar
information about risk management options and their
impact, then we can optimize our decision-making.
Mulkey discussed the development of a risk
management evaluation, a summary of what is
known about risk management options when a
decision needs to be made. It includes an analysis of
sources of risk, the risk management options that
(Continued from page 1)
exist, and the availability, cost, and effectiveness of
those options.
McKinnon said, "It was not our intent to
second guess and rehash the risk assessment, it was
to look at the risk management side. But in order to
do that, we need to provide at least a capsule
summary of what we know about risk. The goal is to
reduce uncertainty in risk management for the EPA
and others in order to provide cost effective risk
management options."
Elements in risk management evaluation include
identification, source strength, timing, and emission
factors. Risk management options include pollution
prevention, source control techniques, management
practices, remediation, resorption, and adaptation.
Resources:
Greg Sayles: 513-569-7607 /
sayles.gregory@epa.gov
Lee Mulkey: 513-569-7689/Mulkey.lee@epa.gov
Hugh McKinnon: 513-569-7689 /
mckinnon.hugh@epa.gov
European Community Risk Management Strategy
Kathryn Tierney, Principal Administrator with
the European Commission (EC), called in from
Brussels with a report on, "European Community
Strategy for Endocrine Disrupters: Implementation
to Date." She said the basic strategy towards EDCs
was adopted by the EC in 1999 and consisted of 3
elements: identifying substances, research & devel-
opment, and legislative action.
Tierney stressed that the strategy was not just
imposed on industry. She explained, "We have to
have buy in from industry, health and consumer
protection, and agriculture, and when the commis-
sion comes up with a strategy like this we have
extensive consultations with member states, NGOs
and others.
The core of the EC strategy is the development
of test methods and research. She noted, "When we
adopted this strategy in 1999, the research was still
going in. We are still waiting for agreed test meth-
ods. We wanted to look at what we can do when
we have agreed test methods."
Short-term actions the EC is conducting while
waiting for agreed test methods to be developed and
validated include:
1. Establishment of a prior list of substances for
evaluation of their role in endocrine disruption.
2. The use of existing legislative instruments. Current
legislation now provides 32 main instruments that
can be used to deal with problem chemicals. One is
classification where substances have to be subject to
existing test methods. Even if they have not been
demonstrated to be EDCs, substances can also be
regulated based on their effects as carcinogens or
for causing reproductive disorders. Tierney said,
"Even though we don't have an agreed idea of what
constitutes an EDC, we should still take whatever
evidence there is into account. We don't want to
bury our head in the sand and ignore the evidence
(Continued on next page)
-------
February 2002
Endocrine/Estrogen Letter
E/E Conference Report
"Even though we don't have an agreed idea of
what constitutes an EDC, we should still take
whatever evidence there is into account We don't
want to bury our head in the sand and ignore the
evidence simply because we don't have an agreed
upon test method." — Kathryn Tierney
simply because we don't have an agreed upon test
method."
3. Establishment of monitoring programs. This is not
necessarily doing new monitoring, but looking at
what is already out there.
4. Identification of specific cases of consumer use.
Tierney noted, "Here we sensed that if we found in
the preparatory work specific substances of concern
to consumers we should not wait for agreed upon
test methods to see what we can do to reduce the
risk. I am talking about cases where you have
vulnerable people like children."
5. Information exchange and cooperation with
international colleagues in order not to duplicate the
same work.
6. Communication
with the public.
Tierney said, "This
was important in
1999 when the
strategy was
published and a lot of media reports were turning up
fear."
7. Consulting with stakeholders in order to insert
feedback into the process.
Medium-term action of the EC includes:
1. Identification and assessment of EDCs and the
development of test methods.
2. Research and development
3. Identification of substitutes. Tierney said they put
that as a medium term objective rather than a short
term one because they weren't sure if the substitutes
could result in the same fears as the chemicals being
replaced.
Long-term action includes:
1. Overhaul of the overall chemicals policy.
2. The water framework directive adopted in 2000,
which overhauled much of the EU legislation on
water.
3. Other legislative actions and proposals.
The strategy adopted by the EC was discussed
with the European Council and the European
EC Risk Management Strategy (continued frompage 2)
Parliament. Tierney said the Council, consisting of
the EU member states, stressed the need to apply
the precautionary principle and the need for quick
and effective risk management policies. The parlia-
ment was even stronger to some extent in criticizing
the commission strategy as not being aggressive
enough.
Tierney also talked about EC work on identify-
ing suspected EDCs. The EC hired BKH consulting
in the Netherlands to put together a report of
chemicals named by Greenpeace, Germany, US,
and the scientific literature, They came up with an
initial list of 553 substances.
Tierney said, "When the study was launched, it
was not expected we would
come up with over 500
chemicals. Within that study
we had to whittle down and
focus on high production
highly persistent substances.
So we gathered more data
on those and finally looked
at exposure."
Quite a bit of misinformation was spawned by
this effort. For example, it was widely reported that
the EU had established a priority list of EDCs.
Tierney said, "This is not the case. The second part
of the process was to consult widely on the list. We
have a commitment to ecotoxicity. We consult with
the World Health Organization and the OECD to
decide what we should focus our resources on."
The actual results of the BKH report were
seen as a reasonable starting point. Tierney noted,
"We were looking at a starting point where he have
to further evaluate the substances. We are only at
the beginning."
The focus of the list was on manmade chemi-
cals and synthetic and natural hormones present in
the environment. All 553 substances were retained
for further evaluation plus 9 natural/synthetic hor-
mones. None have dropped off the list. Tierney said,
"We need a mechanism to update the report so as
we get to know more and new evidence comes in,
(Continued on next page)
-------
Endocrine/Estrogen Letter
February 2002
E/E Conference Report
EC Risk Management Strategy
some will drop in concern and others will rise in
(Continued from page 3)
concern.
An important part of the EC's strategy is to not
duplicate existing community legislation. When they
looked at high production and highly persistent
chemicals, they found that 118 had evidence or
potential evidence of ED.
109 of these were already subject to existing
regulatory measures.
16 were obsolete plant protection products.
14 were already banned from the EU.
22 were subject to marketing restrictions in the
EU.
11 were subject to emission controls.
46 were on priority lists for risk assessment. 31
of these are being risk assessed currently and 15 are
plant protection products or pesticides that are also
being risk assessed.
Tierney said, "We feel that it is an important
result that these substances... are already on black
lists for other reasons. For communications this is
important. Even if we don't know everything about
the ED ability, many are already blacklisted."
None of these chemicals drop off the list.
Tierney explained, "When we do have agreed test
methods, we may decide the some of the substances
need a closer look. That is why none of the candi-
dates dropped off the table."
Following the consultation process and the
result of this report, the EU decided to get away
from the idea of coming up with a priority list of
substances. The main reason is that the EU legislation
has a strict connotation of priority and risk assess-
ment. There is no EU legislation to address EDCs
specifically. So they moved away from a priority list
of chemicals and came up with a priority list of
action.
Out of the other 435 chemicals, they found 12
substances that were not addressed by legislation for
any reason. The EC launched a study of these in
November 2001 that goes through May 2002 to
look for any evidence that they are EDCs. The EC
has also launched a second study of all of the 435
substances with insufficient data in the BKH report
in November 2001 that will run through August
2002.
The EC decided to invite member states to
take EDCs into account during risk assessment and
invited them to carry out classification of 2 sub-
stances they found that were not classified. Tierney
said this means they can carry out or make a
request that these two substances be tested with
existing test methods.
Tierney also discussed the R&D initiatives
being sponsored by the government. The 4th
Community Framework Programme (CFP) for
R&D (1994-1998) spent 8 million euro ($7 million)
on endocrine disrupters, which complemented other
money spent by industry. The 5th CFP for R&D
(1999-2002) budgeted 10.5 million euros ($9.2
million). There is also another 10 million euros being
spent on substances suspected of being EDCs like
PCBs, but the focus is not on endocrine disruption.
The 5th CFP includes another 20 million euros to
be awarded in 2002 towards suspected EDC
research, which does not include industry contribu-
tions. These figures only include European Commu-
nity funding and don't include money going into
national programs in Denmark, Germany, and the
UK.
Legislative strategy is being looked at by the
EC as a long-term action. The overall chemical
policy in Europe is being overhauled. The old
system allowed existing substances to be authorized
for use unless specifically forbidden. Last February,
a white paper came out advocating that substances
that are carcinogenic, mutagenic or reprotoxic
(CMR) would be forbidden unless specifically
authorized. The existing substances include over
30,000 chemicals with annual production greater
than 1 ton. Rigorous tests for long-term health
effects were recommended for chemicals produced
(Continued on next page)
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EC Risk Management Strategy
in quantities greater than 100 tons. Commentary in
the white paper on endocrine disrupters themselves
highlighted the need for further research on endo-
crine disrupters, test methods, low dose effects and
Quantitative Structural-Analysis Relationships
(QSAR).
The white paper was adopted by the EC and
the council adopted the conclusions in June. One of
the things they highlighted is that they know that
EDCs should be subject to authorization. Now the
EC has established various working groups where
they are looking at endocrine disruption. The
commission now has to prepare draft legislation
where they have to give advice on how the EU will
incorporate EDCs into the chemicals policy.
Tierney said, "Up until now, we have had a
parallel EDC process, but the goal is that anything
that we do with EDCs should be consistent with the
overall chemical policy."
The original water framework directive was
adopted in 2000. Tierney said that the EC has
adopted a list of 33 priority substances, and of these
11 are candidate EDCs. When they identified these
priority substances, any evidence of endocrine
disruption was considered, but there were many
other very good reasons to include them. Measure-
ments have to be reported back within 2 years with
the aim of ending or phasing out emissions, dis-
charges and losses within 20 years.
Another piece of relevant legislation is the
General Product Safety Directive. Tierney said,
"This is the only instrument we have that can be used
from an emergency point of view in the short term."
It has been used to create short-term restrictions on
the use of phthalates in toys.
Tierney noted, "We have placed a special
focus on EDCs with this strategy and that has
allowed us to increase the attention given to it in
existing legislation. The EU parliament has recom-
(Continued from page 4)
mended banning EDCs. But the commission is
making sure the whole process is taken up in the
existing process and will be part of the overall
chemical policy rather than proposing new legislation
to deal with EDCs."
Tierney concluded, "The main thing we need to
push is for agreed test methods. Our strategy is that
until we have agreed test methods, we cannot take
legislative action to reduce the risk. In the mean
time, if we come up with some specific cases, then
we can take specific action and use emergency
measures for a temporary withdrawal. But that is a
specific case for consumer use. Without agreed
upon test methods, we can only act on a case-by-
case basis.
William Owens, with Proctor & Gamble, noted
that we have known that many endocrine mecha-
nisms produce reproductive and developmental
effects and that Europe is talking about an authoriza-
tion system for reproductive and development
toxins. He then asked, "How are you going to work
a subset of endocrine disrupters into the system so
you don't have duplication and conflicts and incon-
sistencies?"
Tierney responded, "It is true we have CMRs.
We don't do risk assessments. We just place
restrictions on consumer use. There is a fair amount
of controversy over whether EDCs need a hazard
category of their own. We know that most of the
effects are on the reproductive system. There are 2
things. One is whether the substances have been
through this. That means that the instructions we
now have will have to be relooked at. Also there is a
possibility there are other effects."
Resources:
http://www.europa.eu.in1/comm/environment/docum/
99706sm.htm
Kathryn Tierney: 011 -322-296-8118 /
Kathryn.Tierney@cec.eu.int
Correction: Last months E/E Letter should have reported the number of animals used for endocrine disrupter
testing could potentially climb to 96 million animals if all approximately 80,000 chemicals under consideration
for screening are tested.
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Effects of EDCs On Wildlife
Dr. Gary Ankley, Chief of the US EPA's Min-
Continent Ecology Division, Toxic Effects Charac-
terization Branch, in Duluth, MN discussed an
"Overview of Effects and Assessment of Endocrine-
Disrupting Chemicals in Wildlife." He said there are
a number of implications of EDC effects in wildlife.
There are a number of populations that are experi-
encing serious decline for which we don't have a
cause. It is also theorized that EDCs can result in
community level effects by affecting key species.
The indirect implications are that EDCs can result in
economic issues like reductions in fishing and that
these animals could be acting as sentinels for human
health effects.
But Ankley cautioned that we should not jump
to conclusions that all wildlife deformities are caused
by EDCs. For example, a lot of people are con-
cerned about the implications of deformed frogs on
their children's health. But he said there is strong
evidence that some of these effects are caused by
other factors such as parasites or UV radiation
rather than EDCs.
With a few notable exceptions, such as DBS,
documentation of EDC effects in humans is rare.
Ankley said this is not true in wildlife. There is
evidence of EDC effects on invertebrates, fish,
frogs, reptiles, birds, and mammals.
EDCs have been suspected in causing:
-Hermaphrodism in gastropods
-Feminization or masculinization in fish
-Malformations in amphibians
-Feminization of alligators
-Developmental lethalities/abnormalities in
great lakes fish and birds
-Feminization of mammalian carnivores like
mink and otters
There is evidence that tributyltin is causing
imposex (the simultaneous presence of male and
female genitalia) in gastropods such as snails. This
resulted in sterility and marked declines for specific
populations. It appears the metabolic pathway being
affected is the aromatase enzyme used for convert-
ing testosterone into estradiol. Ankley noted, "What
is really neat about this is that TBT levels are going
down and we are starting to see the population
going up, so we have a complete case history with
regards to these chemicals.
With regards to fish, the evidence started
coming in the early 1990s from John Sumpter's
group in the UK that fish gonads had both male and
female characteristics. Ankley said that since the
initial discovery in the UK, there have been a lot of
studies on municipal discharges in the UK, US,
Germany, and other countries.
Ankley noted that at least in the UK, the
changes seem to be due to natural and synthetic
estrogens such as estradiol. He said, "Here we have
an interesting situation where our pharmaceutical
products could be causing this. There has also been
a focus on alkylphenols. I don't think that is the case
in municipal effluents, but in the case of industrial
effluents, it could be more common."
Another problem that has been noted is the
masculinization of fish near the effluent of pulp and
paper mills in the US, Canada and Scandinavia.
Ankley said it looks like these are caused by
phytoandrogens that are normally bound into the
wood, and then released during paper manufacture.
This is because the same androgenic effect has been
seen at several mills independently of the underlying
chemistry used to make the paper.
Ankley discussed assays they have been
working on in Duluth that could be used for people
involved in risk management decisions of EDCs.
One is the short-term reproductive assay of the
fathead minnow, the "White Rat" of aquatic toxicol-
ogy. The strengths of this assay are
-There are standard methods for culturing and
holding them at different life stages,
-There is a history of use in the regulatory
community
-They are easily handled and cultured
-They are small but still large enough to collect
various tissues from
-They have a relatively rapid life cycle
-They are serial spawners (females spawn
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Effects of EDCs On Wildlife
every 3 days) so you can get an estimate of fecun-
dity.
-They are a dimorphic species (the male is
bigger and has a darker color)
Ankley said they want a short term, cost
effective assay that could be successfully imple-
mented by a range of labs. They also wanted an
assay that would detect and discriminate among
EDC modes of action of concern: estrogen/anti-
estrogen, androgen/anti-androgen, and modulators
of sex steroid metabolism. They also wanted to
incorporate measurements of fecundity, fertility,
hatch, and early survival.
Key endpoints from an EDC perspective
include secondary sex characteristics such as
gonadal weight, plasma vitellogenin, and plasma
steroids.
Most exposure routes occur by adding a
chemical to the water, diet, or injection. Ankley said
injection is a little tricky to do at first, but it is very
effective when you are working with small quantities
of expensive test chemicals, of if you don't want to
release them into the environment.
In developing a particular assay, it is important
to run it through a variety of known chemicals to
establish a baseline level of effects. Ankley recom-
mended:
Beta estradiol - Estrogen Receptor (ER) agonist
Methoxychlor - ER agonist
ZM 189,154 - ER antagonist
Trenbolone - Androgen Receptor (AR) agonist
Methyltestosterone - AR agonist
Vinclozolin - AR antagonist
Flutamide - AR antagonist
Fadrozole - inhibitor of steroid metabolism
Trenbolone is a compound used in animal
feedlot operations to build muscle mass. A recent
Environmental Health Information Service report
noted that much of it passes through the animal and
has a half-life on the order of 260 days, so it could
be involved in runoff. Vinclozolin is an organochlo-
rine fungicide. The parent chemical does not create
an effect but 2 of its metabolites do. Fadrozole is a
(Continued from paged 6)
chemical used to treat breast cancer. It appears to
bind to aromatase, the enzyme that converts test-
osterone to estradiol.
Ankley concluded, "We feel we have a system
that effectively identifies and discriminates among the
EDC modes of concern. Another important aspect
is that it enables routine collection of data (fecundity,
fertility, and hatch) of utility to higher tier risk assess-
ments. The assays being developed applied to both
the single chemical and complex mixture testing."
David Lattier, with the US EPA discussed the
"Development of Biological Methods to Character-
ize Exposure of Wildlife To EDCs." He said they
have developed a test for measuring estrogen in the
field based on gene expression. Most of the work
was done with single gene indicators using single
chemicals to look at these genes and when they are
activated. The initial work focused on fathead
minnows and the expression of the egg protein
vitellogenin.
The research used estradiol, ethinylestradiol,
and DEHP to cause the gene expression. When the
estrogen enters the cell membrane, it binds with the
estrogen receptor, resulting in gene activation.
Lattier said the test has identified vitellogenin at
levels of chemical as low as 2 ng/1 within 8 hours of
exposure. By looking for the gene rather than the
expressed vitellogenin, they are able to attain a
higher level of consistency. Lattier said the standards
of deviation are considerably higher for the detection
of the protein than for the gene that makes the
protein.
Lattier said that his group wanted to put the
single gene test into the hands of the EPA regions.
He noted, "We will do a demonstration in your
backyard, but you must provide the food."
Myriam Medina-Vera discussed the "Develop-
ment of Chemical Methods to Characterize Expo-
sure to EDCs in the Neuse River Basin" in North
Carolina. She said the Neuse River basin is a highly
industrial area with about 1.5 million human inhabit-
ants and a substantial hog population.
Some of the suspected EDCs being studied in
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February 2002
E/E Conference Report
Effects of EDCs On Wildlife
the region include alkylphenol polyethoxylates,
antibiotics, pesticides, and metals including arsenic,
cadmium, copper, manganese, lead, and tin.
She said the alkylphenol polyethoxylate
research is a hot area and that a lot of countries are
trying to decide whether to regulate them. Some of
the variants are persistent, accumulate in fish, and
can link to the estrogen receptor. They are used in
making pulp and paper, plastics, leather, agriculture,
household, industrial and institutional cleaning
products, and as a spermicide in contraceptive
jellies and creams. The US is one of the largest
producers and consumers.
The degradation mechanism of these chemi-
cals is of interest, because Medina-Vera noted,
EPA'sEDC Strategy
(Continued from page 7)
wastewater treatment plants are only capable of
degrading 30-35% of the nonylphenol. She said the
controversy is that the combination of the
monoethoxylates and acetic acid congeners maybe
more toxic than nonylphenol. She pondered, "How
do you define the toxicity of chemicals you are
looking at. The first question they are going to ask
you is which ones are the toxic ones, which are the
ones you really want to control."
Resources:
GaryAnkley:
218-529-5147 / ankley.gerald@epa.gov
Myriam Medina-Vera:
919-541 -5016 / medina-vera.myriam@epa.gov
Elaine Z. Francis, National Program Director
for Endocrine Disrupters Research at the US EPA,
spoke about "EPA's Endocrine Disrupters Screen-
ing Program: Legislation, Implementation, and
Research." She outlined the basis for the EPA's
EDC mandate from the 1996 Food Quality Protec-
tion Act and Safe Water Drinking Act. These
specified that the EPA must develop a program in
August 1998, implement the program in 1998, and
report to congress in 2000, although the schedule
has slipped considerably. The mandates did provide
some discretionary authority. For example, the EPA
can require screening and testing of:
-Any active or inert pesticide ingredient
-Any chemical that has an effect cumulative to
an effect of a pesticide
-Drinking water source contaminant
- Other endocrine effects
- Environmental effects
A snapshot of the chemicals that need to be
looked at include:
-900 pesticide active ingredients
-Inert ingredient found in various combinations
with pesticide active ingredients
-Approximately 75,500 industrial chemicals in
the Toxic Substances Control Act. This does not
include metals or sanitation byproducts.
The proposed phase 1 of the testing program is
to focus on pesticide active ingredient and high
production volume chemicals. Francis said, "It is a
fairly reasonable size of chemicals (1500-1600) to
begin screening."
There are a number of factors that go into
deciding the priority of a particular chemical such as
persistence, presence in sediments, presence in food
and water, andbioconcentration. Francis noted,
"For the most part, we lack a lot of information in
terms of the effects for this priority setting. There are
two solutions we have come up with."
One approach was to use High Throughput
Screen (HTPS) technology. The EPA thought it
would run thousands of chemicals through to see
where they fall for the remainder of the Tier 1 screen.
Francis said, "We did do HTPS, but felt that it was
unreliable for regulatory use. We are now focusing
on the second solution, QS AR." However, both
approaches need further validation, which is man-
dated by law.
Francis said Tier 1 validation is under way, and
should be done by 2004, when Phase 1 of testing
will begin. Tier II validation will continue through
2005.
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Exposure Assessment of Suspected EDCs
One of the missing elements in the debate over
EDCs is data about the levels of chemicals humans
and wildlife are exposed to. Although many chemi-
cals demonstrate some ability to affect hormones,
there is not much cause for concern if exposure is
low. A number of researchers presented information
about research in this area.
Steven Goodbred, with USGS talked about
"Monitoring Endocrine Disrupting Compounds In
Aquatic Ecosystems in the US." He said there is no
systematic knowledge of EDCs in the US due to 1)
an incomplete consensus on EDCs, 2) analytical
methods are still
being developed
for many com-
pounds, 3) difficulty
selecting a sampling
matrix, and 4)
major funding is needed.
Several agencies have national monitoring
programs that include EDCs. The USGS has
NAWQA, TOXICS, and BEST. The EPA has
EMAP, REMAP, and the National Dioxin Study.
NOAA has Mussel Watch and Benthic Surveillance.
The FDA has the National Monitoring Program for
Food and Feed.
The USGS launched the National Water
Quality Assessment Program (NAWQA) to assess
the status and trends of surface and ground waters
throughout the US. It surveys 60 basins throughout
the US and cycles through them every 4 years.
Goodbred said, "We are looking at factors that
affect water quality including EDCs."
In the urban water samples analyzed, they
found that 100% of the fish had pesticide exposure
and 99% had some pesticides. In agricultural areas,
85% of the fish had been exposed to pesticides and
92% of the water samples contained them. 33% of
the major aquifers tested positive for pesticides.
Pesticides were found to almost always occur
as mixtures. About 50% of the sites tested had at
least 6 different pesticides. Goodbred said, "Organ-
isms are exposed to very complex mixtures, and this
NAWQA Study Results
Matrix Suspected EDCs
Water for hydrophilic pesticides 20
Bed sediment for PAH and phenols 17
Fish Tissue for organochlorines and PCB s 24
is something we need to deal with for risk assess-
ment."
One issue is that pesticides in streams usually
occur in strong seasonal pulses. Goodbred noted,
"If you are not out there sampling daily during storm
surges, you are going to miss a lot of information."
Although the average concentrations are far below
the maximum levels allowed, individual peaks are
much higher.
Another issue is that the breakdown products
can be 10-25 times the concentration of the parent
compound. Goodbred said, "We really need to
understand the
kind of break-
down products
and whether they
are endocrine
disrupters. That
could keep chemists busy for the next millennium."
Goodbred concluded, "It appears a lot of
EDCs are potentially widespread in the US. That is
not a surprise, but it is interesting and something we
need to deal with for risk assessments."
The USGS is preparing to release the National
Reconnaissance of Pharmaceutical and Other
Emerging Contaminants in US Streams this coming
March. The study will try to determine if pharma-
ceuticals, antibiotics, hormones and other chemicals
consumed by humans and animals are entering the
environment. The approach is to develop sensitive
and specific analytical methods for 22 antibiotics, 14
prescription drugs, 5 non-prescription drugs, 15
hormones and steroids, and 39 household and
industrial compounds.
Phase I was a reconnaissance of 13 9 streams
in 30 states broken down into agriculture, urban,
mixed land use, and minimally developed groups.
The preliminary data indicated that 82 out of 95
wastewater compounds looked for were found in
80% of the stream samples. Detection of multiple
compounds was common. About 75% had more
than one, and about 35% had more than 10. The
steroids had one of the highest concentrations, but
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Endocrine/Estrogen Letter
February 2002
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Exposure Assessment
nonylphenol and its ethoxylates also ranked high.
Goodbred reiterated that the USGS has a lot
of quality-controlled data, and is willing to make it
available to anyone who wants it free of charge.
Ruthann Rudel, a senior environmental toxi-
cologist at the Silent Spring Institute discussed
"Residential Indoor Air and Dust Measurements of
Phthalates and other Endocrine Disrupting Com-
pounds." Rudel said, "There are a large number of
lexicologically important chemicals that are wide-
spread in indoor environments."
She has been involved in an epidemiological
study of 2100 people in Cape Cod, MA. As part of
that study, her team also been looking at the levels of
90 suspected EDCs found in blood, urine, dust, and
air in a subset of 120 of the test subjects. She
presented data on 30 of these subjects. The subjects
were primarily older women over 65 years old. It
included individuals who reported both low and high
uses of pesticides.
Rudel said exposure assessment is a weak
element in environmental epidemiology, and that the
government accounting office has issued a couple of
reports citing this weakness. Exposure data could
help to identify highly exposed populations.
The preliminary data showed the most abun-
dant chemical in the air was o-phenylphenol. Other
common chemicals included Di-Butyl Phthalate, Di
Ethyl Phthalate, pentachlorophenol, diazinon,
heptachlor, methoxychlor, folpet, piperonyl, butox-
ide, carbaryl, permethrin, and chlordane. Rudel said,
"I was interested in how many banned pesticides
were still in air samples."
They also found that indoor air concentrations
of suspected EDCs were sometimes 100 times
(Continued from page 9)
greater than in outdoor concentrations. The team
also decided to look at dust, which Rudel described
as, "a good reservoir reflecting current and past use
of chemicals in the house." Dust also presents an
exposure pathway for children. They found phtha-
lates are clearly the most abundant in dust. Pesti-
cides found in the dust included chlordane, dieldrin,
and DDT.
They also looked at EPA Preliminary
Remediation Goals for available compounds and
compared these to the results. In general the
samples were below these levels with the exception
of chlordane, heptachlor, and PCB52.
Rudel said a striking thing about the results was
that the profile in urine and air samples looked
similar to each other. "If you take these exposure
assessments, you would not find that these air
concentrations would provide any significant dose.
One hypothesis is that the ambient concentration
indoors could be proxies for various exposures,
such as putting on hair spray."
Rudel concluded her presentation, "We are
currently beginning the analysis for our full set of
data. We are looking forward to interesting and
informative findings and we are seeking support for
additional work and collaboration in this area."
Resources:
Nawqa: http ://water.usgs .gov/nawqa/
National Reconnaissance of Emerging Contaminants
in the Nation's Water Resources:
http://toxics.usgs.gov/regional/emc.html
Steven Goodbred:
916-278-3097 / goodbred@usgs.gov
Ruthann Rudel:
617-332-4288 / rudel@silentspring.org
Rumor Mill
A source told E/E Letter that legal action might be forthcoming around a case in which Frederick Vom Saal
alleges Dow Chemical tried to bribe him to suppress data around the low-dose effects of Bisphenol-A.
However, Vom Saal responded, "I wrote a letter to the FDA about it, but I have no desire to spend any
more time pursuing this case."
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^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^H
Risk Management Approaches
Greg Sayles, a chemical engineer with the US
EPA's National Risk Management Research Labo-
ratory discussed "EPA's Risk Management Evalua-
tion (RME) of EDCs." An RME summarizes the
current understanding of risk management of a
particular environmental challenge and includes data
about known health and ecological effects, signifi-
cant sources to the environment, and significant
exposures and environmental sinks. The RME on
suspected EDCs includes chapters on a number of
chemical classes including alkylphenol ethoxylates,
DDT and DDE, natural, veterinary, and pharmaceu-
tical steroid hormones, PCBs, chlorinated dioxins,
andfurans.
The RME for EDCs identifies currently avail-
able risk management approaches that were devel-
oped for other purposes, but appear useful in
managing the risk of EDCs. The document also
indicates where new risk management approaches
are needed. The RME will be useful 1) to inform
risk managers such as regulators on what technical
approaches are currently available for managing
EDC risk, 2) to educate the public about what risk
management approaches are and are not available
now, 3) to motivate environmental consultants/
engineers to review current skills or to develop new
skills applicable to managing exposure to EDCs, and
4) to guide risk management researchers, such as
NRMRL, in planning EDC risk management re-
search programs.
After the presentation, Mark Walton, Business
Public Issue Leader, at Dow Plastics noted that
these kinds of lists from a regulatory agency could
be exciting to the media because they can be taken
out of context. He asked how the EPA planned to
communicate information about these suspected
EDCs.
Sayles responded, "When you see the docu-
ment you will see a fair amount of caveats around
that list. They are very practical caveats. You cannot
work on every known chemical forever and try and
get a document done. We are very clear these are
compounds still under study. We have made that
very clear."
Vincent Kramer, with Dow AgroSciences
commented, "Every time you consider a substitute
for a product, you need to consider the risk of
removal of that benefit. In the area of growth
promoters for cattle, it could be argued that they
help supply a meat product that is affordable to the
general population that has benefits for nutrition. If
you consider another way of growing cattle it may
not include the fact that it may make the price of
meat unaffordable."
Carolyn Acheson, a chemical engineer with
NRMRL discussed "Using Bioassays to Evaluate
The Performance of Risk Management Techniques."
She started the presentation off noting, "One lesson
I have learned is that you will have a much better
understanding of your risk management technique if
you include bioassays."
She said researchers in this area make simplify-
ing assumptions that ignore important factors. A
common assumption is that if we know the contami-
nant, we understand the risk. But a researcher may
ignore the fact that there is an incomplete removal
of, or the development of side products or that there
are co-contaminants.
Another common assumption is that treatment
reduces toxicity. But a researcher may ignore
process amendments, other reactions or matrix
changes. For example, they have learned when
remediating gasoline that if you aerate the soil you
can reduce contaminant concentrations, but you can
also release metals.
If you are just using chemical assays to look for
changes, you might miss these important side effects.
By integrating a bioassay into the remediation
process, you can notice these side effects, and then
use chemical assays to pinpoint the chemicals and
processes responsible.
Acheson described a case study where they
were looking at remediating PCB from soil. After
treatment, the levels of PCB were reduced, but a
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(Continued from page 11)
bioassay revealed the soil was still quite toxic. It
turned out that in the course of reducing PCB
concentration, they had created propanol. So they
added a rinse step, which reduced the total toxicity
of the soil.
Four types of bioassays that the NRMRL has
been exploring include: 1) sediment/aquatic inverte-
brate tests, 2) terrestrial invertebrate tests, 3) in-
vitro tests and 4) seed germination and root elonga-
tion. The sediment aquatic research test uses a
commonly studied aquatic organism but requires
substantial lab equipment and takes about 1 month.
The terrestrial invertebrate test uses an earthworm
model. It has disadvantage that it cannot measure
imposex effects because the test is not sensitive to
androgens. The in-vitro assays include an e-screen
based on mouse breast cells lines and yeast cells.
Resources:
Greg Sayles: 513-569-7607 /
sayles.gregory@epa.gov
Carolyn Acheson 513-569-7190 /
acheson. carolyn@epa.gov
Drinking Water Treatment
An important area of risk management for
confirmed and suspected EDCs lies in removing
them from the drinking water. If research indicates
that existing treatment processes can ameliorate
these chemicals sufficiently, then perhaps no other
steps will be necessary. But if they don't, new
processes will have to be developed for dealing with
these chemicals, particularly if they are detected at
high levels.
John Cicmanec, working with the National
Risk Management Research Laboratory reported
that a number of treatment processes had been
tested for their efficacy in removing chemicals such
as alkylphenols, bisphenol-A, phthalates, PCBs,
dioxins, dibenzofurans, and the pesticides methoxy-
chlor, endosulfan and atrazine. Cicmanec said a
number of other vectors are on the horizon of
concern including coxsackievirus (juvenile diabetes),
heliobacter pylori (thyroid disorders), trenbolone
acetate, andmelengestrol acetate.
In addition to the conventional water treatment
process of sedimentation, coagulation, and filtration,
they also considered the effects of granular activated
carbon (GAC), powdered activated carbon (PAC),
nanofiltration, reverse osmosis, and air stripping for
the removal of confirmed or suspected EDCs. Their
findings indicate that GAC and PAC were the most
effective at removing the chemicals tested.
Cicmanec spoke about nonylphenol noting that
after Sweden established regulations, nonylphenol
rates decreased from 75 micrograms/1 down to 9
micrograms/1 within one year. He added that
nonylphenol is found in 24% of the water samples
from the US.
Looking at traditional water treatment tech-
niques, Cicmanec said they found that by simply
performing coagulation, they were able to remove
87% of methoxychlor and 98% of DDT. Other
chemicals were harder to remove. For example, the
traditional techniques only removed from 10-60% of
lindane, aldrin, dieldrin & parathion. They also found
that UV, which is used to kill microbes, might also
have an effect in helping to break down chemicals.
PAC and GAC are the most effective tech-
niques observed to date, but their costs keeps them
beyond the reach of many plants. PAC is used in
about 48% of the surface water plants, and 12% of
the ground water plants. GAC is used in 12% of the
surface water and 5% of ground water treatment
plants.
Factors that affect the effectiveness of PAC/
GAC include contaminant adsorbability, concentra-
tion, multi-component adsorption, adsorption
kinetics, temperature (works best at low temp)
carbon dose, and contact time.
In Cincinnati they added a PAC plant a few
years ago at a cost of about $60 million, which
amortizes out to a present cost of 27 cents/1,000
gallons versus 25 cents/1,000 gallons before.
Kathleen Schenck, an environmental scientist
(Continued on next page)
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February 2002
Endocrine/Estrogen Letter
13
E/E Conference Report
Drinkin
with the NRMRL discussed "Evaluating Drinking
Water Treatment Technologies for Removal of
EDCs. She said two basic strategies exist for
decreasing the risks of potential adverse health
effects from EDCs in drinking water. One is to
protect source waters from contamination by EDCs.
The other is to remove EDCs during the treatment
process.
Schenk's team evaluated the removal of a
number of compounds including estradiol, estriol,
ethinylestradiol, progesterone, testosterone, and
dihydrotestosterone. The project was divided into
four parts. The first is the development of an analyti-
cal method to identify and quantify the analysis. The
second is the application of a reporter gene MVLN
assay to evaluate the removal of estrogenic activity
from the water. It is designed to detect compounds
that may have been created in treating the water.
The third part is the use of bench scale evaluations
of various drinking water processes. The final part
will be to conduct pilot scale evaluations of these
techniques, if warranted.
The proj ect was launched in November 2000
and is still in process. Schenck said she had ex-
pected to see some engineering data by now, but
the analysis is taking longer than expected. The
steroid hormone analysis should be done within a
year. Work has not even started on the nonylphenol
compounds.
Schenck noted that at the moment, there are
no initiatives to regulate any of the chemicals being
tested. She said, "We don't even know if there is a
human health risk associated with the presence of
them. The reason we are doing this is that NRMRL
has decided not to wait until every last decision is
made on a risk assessment, but to go ahead and
look at emerging problems and how to fix them." It
may turn out that GAC is all that is required.
Fred Pontius, President of Pontius Water
Consultants, in Lakewood CO discussed "Risk
Management of EDCs in Drinking Water." He said
there are four simple questions at the heart of the
matter:
g cl (Continued from page 12)
1. Are current regulations adequate?
2. Should we be concerned about unregulated
EDCs such as pharmaceuticals?
3. What can water utilities do to manage risks?
4. What are the critical research questions?
Drinking water contamination is currently
regulated by the Safe Drinking Water Act. Pontius
said that we may not be able to regulate based on an
endocrine disrupter effect, but in many cases,
chemicals are covered by other types of effects. The
EPA also has the latitude to monitor unregulated
compounds of concern.
When a chemical is regulated, the EPA sets a
maximum containment level (MCL) goal. But in
1996 the EPA was directed to determine if the
benefits of an MCL justified the cost. Pontius said,
"This is not going to be a straightforward proposi-
tion. We are learning that with arsenic."
Pontius believes that overall, current regulations
are sufficient, even though the EPA has not started
down the path of regulating EDCs.
With regards to unregulated contaminants,
Pontius noted, "Sometimes we find them, but most
of the time we don't. We need to recognize there
are many types of chemicals and considerations. It is
important to be careful with generalizations. If a
regulation has been set that requires me to install a
treatment system, I will not target the one chemical. I
will try and get as much out of that as I can."
The critical research questions in this area include:
1) What are the relative health effects of EDCs to
determine which chemicals are important to remove?
2) What is the occurrence, fate and transport of
EDCs?
3) What is the treatment effectiveness on EDCs?
Resources:
JohnCicmanec: 513-569-7481 /
cicmanec.john@epa.gov
Kathleen Schenck: 513-569-7947 /
schenck.kathleen@epa.gov
Frederick Pontius: 303-986-9923 /
fredp@pontiuswater.com
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14
Endocrine/Estrogen Letter
February 2002
E/E Conference Report
Concentrated Animal Feed Operations
A major source of environmental hormones are
confined animal feed operations (CAFO) in which
thousands of animals are housed, sometimes with
substandard sewage treatment systems. The quantity
of hormones and hormone mimics is increased by
the use of hormones and antibiotics to increase
growth, and substantial populations of pregnant or
lactating animals. Researchers presented information
on a number of different aspects of the concerns
about how to manage the risks of these CAFOs.
Steven Hutchins, a research environmental
scientist with the US EPA National Risk Manage-
ment Research Laboratory presented a talk on the
"Potential of CAFOs to Contribute Estrogens to the
Environment."
There are many unknowns regarding the
potential for CAFOs to contribute EDCs to the
environment, due to the variety of CAFO opera-
tions, the diverse natures of potential EDCs them-
selves, and the different types of endocrine recep-
tors that could be affected. Two projects have been
initiated. One, to evaluate EDC activity from differ-
ent types of CAFOs and the other to measure levels
of estrogens in swine waste effluents.
The first project is being conducted by Okla-
homa State University under a cooperative agree-
ment and is designed to evaluate swine, dairy, and
beef CAFO lagoons. Three analyses will be used to
assess for the presence of EDCs: 1) Xenopus Tail
Resorption Assay to measure for thyroid disrupters,
2) Enzyme Linked Immunoassay (ELISA) to detect
estrogenic compounds in frogs, 3) A measurement
of plasma testosterone and 17a-estradiol concentra-
tions as an indicator of alteration in reproductive
endocrine homeostasis.
The work is ongoing. Preliminary results
indicated that although the swine effluent is quite
toxic, none of the lagoons exhibited estrogenic
activity.
In the second study, Man Tech Environmental
Services is working under contract to optimize both
ELISA and Liquid Chromatography/Mass Spec-
trometry/MS analytical procedures for the analysis
of complex wastewaters. The goal is to use ELISA
for screening of environmental samples for estro-
genic activity, and then to confirm the presence of
the individual estrogens in samples that test positive
withLC/MS/MS.
The research is ongoing. Preliminary results
show the analysis works well for ground water
samples, with ELISA detection limits on the order of
.05 ng/1 and 2 ng/1 for estrogen separation and
identification by LC/MS/MS. One problem is that
ELISA often yields 17a-estradiol concentrations
orders of magnitude higher than what can be con-
firmed by LC/MS/MS.
Pigs are not typically given growth hormones,
so most of the focus with them is on the natural ones
produced by the pigs themselves. Poultry are not
generally given growth hormones either, although
they still produce quite a bit of natural estradiol.
Hutchins said there are estimates that the US poultry
population produces between 160,000 and 760,000
kg/year of estrogen.
With Cattle however, an estimated 90% are
fed growth hormones including estrogens, andro-
gens, andprogestins. Scientific literature on the
matter revealed an average estradiol concentration
of 10-13 ng/1 in the urine. After administration of
growth hormone, the estradiol concentration is 5-6
times higher.
Suresh Rao, a researcher at Purdue University
discussed "Investigations of Sorption and Transport
of Hormones and Animal Pharmaceuticals: Initial
Laboratory Results." Pigs are big business in Indiana
with a total population of 5 million. Although the
number has remained constant, the CAFOS have
become more concentrated with a reduction in the
number of operations from 25,000 to 4,500 be-
tween 1979 and today. Over 60% of these opera-
tions have more than 2,000 hogs. The concern is
that a large number of pharmaceutical compounds
and antibiotics are used as growth promoters.
A report from the Union of Concerned Scien-
tists claimed that the anti-microbials used in human
health are dwarfed by animal uses for non-therapeu-
(Continued on next page)
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February 2002
Endocrine/Estrogen Letter
15
E/E Conference Report
CAFOs
tic purposes. About 10.3 million pounds/year are
used on hogs, 10.5 million pounds/year on poultry,
and 3.7 million pounds/year on cows, compared to
about 3 million pounds per year on humans. The
animal numbers would be even higher if one were to
consider therapeutic uses as well.
Rao said the initial project was to identify
major growth promoters and hormones present in
swine CAFOs. The major hormones included 17a-
estradiol, 17a-ethinylestradiol, and testosterone.
Growth promoters included carbadox, tylosin,
tetracycline, andbacitracin.
Rao said that he started out thinking that birth
control pills and hormone replacement therapy
would be the largest sources of hormones from
people. But subsequent research revealed that 60%
of the excreted hormones in wastewater come from
pregnant women. The total human load of estrogen
is estimated at 675 kg/year. With a few bold as-
sumptions about wastewater load, Rao estimated
human estrogen excretion results in an average load
of about 300 ng/1.
Another issue is the hydrological transport of
these compounds. Rao said they have a high sorp-
tion coefficient and that the facilitated transport to
water bodies is possible. They are completing lab
experiments to continue to collect data and there are
plans to conduct field experiments to measure what
happens in a field that receives manure applications.
There are plans to go out to the watershed in a large
C AFO and monitor water from the source to
various distances from it.
Helder Hakk, a researcher with the USDA's
Biosciences Research Lab Animal Metabolism-
Agricultural Chemicals Research Unit presented a
talk on the "Fate of the Endogenous Hormones 17a-
estradiol and Testosterone in Composted Poultry
Manure and Their Sorption and Mobility in Loam
Soil and Sand." He noted that estradiol is one of the
most potent hormones and is about 1,000 to
100,000 times as potent as alkylphenols. It is
naturally excreted in the urine of mammals and birds.
Hakk estimated that chickens on the east shore of
(Continued from page 14)
Maryland produce about 6 kg/day of estradiol.
A variety of effects of estradiol have been
noted. At low levels (5-500 ng/1) estradiol increased
alfalfa growth. At higher levels (5000-500,000 ng/1)
estradiol decreased alfalfa growth. Levels of 300
ppb in poultry feed caused premature udder devel-
opment. 1-10 ng/1 increased vitellogenin growth in
fish.
Hakk said, "Up until this point, the main
concern has been nutritive, but little attention has
been on the chemical nature of some of the minor
materials in the manure." He decided to find out if
composting could help reduce the levels of estrogen
in the manure.
For an aerobic composting process, he used a
2-acre site with an 8" thick pad separating the
compost from the dirt. The composting recipe they
tried combined chicken manure with a variety of
commercial products to create a carbon to nitrogen
ration of 3 0:1. They were deposited in 160' x5'
rows, 40" high.
Throughout the composting process, samples
were quick frozen and sent out for analysis. Test-
osterone concentrations started at around 200 ng/g,
and degraded at an average rate constant of .07521
day. The bulk of the degradation occurred early in
the study when the high temperatures were most
active. Estradiol concentration stated at 100 ng/g,
and degraded at arate constant of .0121/day.
Hakk believes there are two rates of degrada-
tion occurring. A rapid degradation during the
thermophilic period, and then a slower degradation
after the compost cools down. In the end, neither
hormone was degraded completely after 19 weeks.
He said, "Composting may provide an environmen-
tally friendly method for reducing, but not eliminating
the introduction of potent endocrine hormones."
Hakk also reported on the results of another
study looking at the transport and fate of testoster-
one and 17a-estradiol in columns packed with loam
soil or sand. Combustion analysis revealed that 80%
of the testosterone and 96% of the estradiol re-
mained in the top 5 cm of soil. The conclusion was
(Continued on next page)
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16
Endocrine/Estrogen Letter
February 2002
E/E Conference Report
CAFOs
that estradiol and testosterone will be readily trans- ing the hormones.
ported through the sand, but they are rapidly and
strongly sorped into loam soil. Preliminary data also
suggests that they may be metabolized in soil.
Future work includes confirming the compost
extraction test results with mass spectrometry. Hakk
would like to experiment with prolonging the ther-
mophilic heated phase by adding extra carbon-
aceous material to see if that works better at reduc-
Wastewater Treatment
(Continued from page 15)
Resources:
Stephen Hutchins: 580-436-8563 /
hutchins.steve@epa.gov
Suresh Rao: 765-496-6554 / pscr@purdue.edu
Heldur Hakk: 701-239-1293 /
hakkh@fargo. ars.usda.gov
Human waste streams contain a number of
hormones and hormone modulators. Greg Sayles, a
chemical engineer with the US EPA's NRMRL said
the most important compounds from a sewage
treatment perspective are natural and synthetic
hormones and alkylphenol polyethoxylate related
compounds. The former are either manufactured
directly by humans or consumed in the form of
hormone replacement therapy and birth control pills.
The latter are often used in soaps and wetting
agents.
A1998 report in Environmental Science and
Technology, by Desbrow et al reported that public
owned treatment works discharged several estro-
genic compounds including estrone (1-50 ng/1) 17
B-estradiol (2-50 ng/1) and 17 a-ethinylestradiol (0-
7ng/l)
Sayles said these chemicals are accumulating in
ecosystems impacted with sewage treatment efflu-
ent, indicating that they are not being adequately
degraded in the sewage treatment plant.
Alkylphenol polyethoxylates have been shown to
lose portions of the polyethoxylate chain during
sewage treatment, however the most prevalent
resultant compound, nonylphenol, is known to
accumulate and cause toxicity in aquatic organisms
even at low concentration. Thus degradation poses
more of a risk than the parent compound. Sayles
said, "There is no evidence that alkylphenols are
destroyed, just transformed."
According to Sayles, over 500 million pounds
of NPE surfactants are used in the US annually and
about half of them end up going into the sewage
system. Estradiol has been detected at levels as high
as 50 ng/1 in sewage. Estriol estrone has been
detected at levels up to 100 ng/1.
Research indicates that some plants do better
at breaking down hormones, but more research is
needed to determine why. Two factors that have
been identified are retention time and nitrification.
Sayles said, "We need detailed studies on the
fate of these EDCs within the plant. We really need
some good study on what happens within the plant
and how variables like temperature affect it. We
should also be thinking about other treatment
systems. Most of the data we have is associated
with the big plants, but a large number of people in
the country use septic systems on their own land or
small community treatment systems."
Paul McCauley, with the US EPA's NRMRL
discussed a pilot plant to study different treatment
approaches of human sewage, and the effectiveness
of each step in wastewater treatment. The long-term
objective is to determine the fate of estrogenic
compounds and to possibly fashion a cure for the
problem.
His team has built two pilot scale wastewater
treatment system to model metabolic pathways to
study aerobic and anaerobic digestion. It uses rabbit
waste mixed with glycerin to simulate fats.
McCauley said, "Our primary objective is to
get a pilot plant running and identify what works. We
need some cursory comparisons to outside plants."
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February 2002
Endocrine/Estrogen Letter
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The Challenges
Combustion processes have been implicated
as a source of EDCs in the air. Brian Gullet talked
about "Endocrine Disrupters from Combustion and
Vehicular Emissions: Identification and Source
Nomination." There are ongoing efforts to analyze
exhaust samples from combustion and vehicular
sources to provide initial identification of EDCs. The
intent of this screening effort is to provide discerning
evidence for nominating sources for further EDC
characterization. Conventional sampling, advanced
analytical methods, and bioassays are being used to
provide initial characterization of these samples for
their compound identity and EDC activity.
Gullet said the intent is to sample and chemi-
cally characterize multiple combustion sources,
consistent with the likelihood that combustion
source EDC exposure is not linked to any one
source. For example, since body burdens of
polychlorinated dibenzodioxin/dibenzofuran do not
appear to be elevated near traditionally suspected
sources like waste incinerators, it appears that
exposure sources of dioxin are ubiquitous.
Sample fractionation will be coupled with
chemical characterization, quantitative structure
activity relationship analysis, and bioassay testing to
nominate and identify potential EDC compounds in
combustion emissions. The ability to provide this
early source-specific EDC identification and char-
acterization of combustion sources will be a parallel
activity with more extensive health risk analysis and
exposure assessment. In this manner appropriate
prioritization of EDC management options can be
implemented prior to definitive health effects conclu-
sions.
Gullet said, "We only see dioxins and furans
because we look for them. Our intent was to over
the next year or so look at several combustion
sources and go into a fairly extensive analysis of
these combustion emissions to try and understand
what EDC compounds are in them."
Gullet's team has analyzed a wide range of
sources over the last year and a half including
of Combustion
domestic waste burning, diesel trucks, forest fires,
fireplaces, wood stoves, wheat burning, and others.
He said, "The idea was to not only sample the
sources, but to also apply bioassays to see which
compounds were potentially EDC active."
His team has developed a novel technique
called multi dimensional gas chromatography-mass
spectroscopy (MDGC-MS), which gives better
results than using traditional gas chromatography-
mass spectroscopy (GC-MS). Many compounds
were found with MDGC that would have been
difficult to detect in conventional GC-MS because
of elution. For example, a methoxy alkylphenol
waste compound was found to coelute with biphenyl
under conventional conditions but was well sepa-
rated using MDGC.
They are planning to use QSARto look at
what compounds they need to focus on. There are
also plans to test more construction fires, which is a
substantial concern because of the estimated
500,000 structural fires a year. Other plans call for
more diesel vehicle studies. The group has built the
world's only diesel vehicle with a built-in system for
sampling dioxin and furan emissions.
Gullet said some of the more interesting work
over the last couple of years has been on emissions
of dioxins and furans from backyard domestic
burning. While this may not be a very common
practice, the emissions per pound are approximately
4-6 orders of magnitude (10,000 - 1 million times)
higher than from a controlled incinerator facility.
Tests from forest fire emissions indicated levels
of dioxins and furans 10 times higher than previously
thought. Gullet said the reason for the discrepancy
was that this was the first time that anyone had
sampled a forest fire for furans. The EPA had
traditionally relied on data based on industrial
combustion of wood for the previous estimates.
Resources:
Brian Gullet:
919-541 -1534 / gullet.brian@epa.gov
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18
Endocrine/Estrogen Letter
February 2002
E/E Conference Report
Cost Effective
PCB cleanup costs can be quite substantial.
For example, GE is spending $ 1757 cubic yard to
dredge the Hudson for PCBs. A much more cost-
effective option is natural recovery, in which the
forces of nature hide the problem with little or no
human intervention. Richard Brenner, an engineer at
the EPA's NRMRL presented a talk on the effec-
tiveness of this technique.
Monitored natural recovery includes careful
assessment, modeling, and monitoring to ensure
success. Processes that contribute to natural
recovery include biological processes, physical and
chemical processes, and sorption. Storm events,
construction and industrial activities can all work
against the process by disturbing the sediments.
A Clemson, SC site was selected because it
had a documented history of contaminated sedi-
PCB Cleanup
ments. Capacitor manufacturers dumped an esti-
mated 400,000 tons of PCBs into the water be-
tween 1955-1978. Battelle collected core samples
at 10-transect locations between 1994 and the
present time.
An analysis of the results indicated that begin-
ning in 1955, PCB levels started to increase until
1977 when operations ceased. Then PCB levels
began to decrease down to about 1.5 ppm. Brenner
said it is estimated it will take another 1 -5 years to
reduce surface concentrations below 1 ppm, and
then another 10-30 years before they reach .05
ppm.
Resources:
Richard Brenner: 513-569-7657 7
brenner.richard@epa.gov
Finding EDC Alternatives
As the various efforts to identify and characterize
EDCs proceed in the US, Europe, and Japan,
manufacturers will eventually be faced with the
prospect of finding alternatives to some of the
chemicals. The danger is that some of the alterna-
tives may be just as bad, if not worse than the
original chemicals. In order to address this issue, the
EPA has launched a program for the identification
and replacement of EDCs.
Douglas Young with the US EPA's NRMRL said
there are two distinct phases of the program involv-
ing 1) identifying potential EDCs, and 2) providing
knowledge on replacing these in current commercial
applications. Young said they would like to incorpo-
rate this knowledge into software that could be used
by commercial and government researchers.
The EPA is looking at using a QS AR model such as
COREPA and COMFA to identify compounds of
concern. Young said they would like to fund some-
one to do a 3rd model that would be complementary
to these other 2 models.
For the second part of the project, they want to
develop a piece of software to suggest replacements
for EDCs in anti-oxidants and surfactants applica-
tions. The EPA has already developed software
called Program for Assisting the Replacement of
Industrial Solvents (PARIS) for chemical solvents.
Young said they would like to repurpose the coding
and framework of PARIS for EDCs. However the
primary properties of anti-oxidants and surfactants
will be different than for solvents. A limitation of this
kind of technology is that it will not work for all
EDCs such as pesticides and herbicides.
The RFPs for this project are due by the end of
February, with the intent of getting the project up
and running by mid fall. Meanwhile, Todd Martin at
the EPA is working on converting the PARIS
program framework to work with EDCs. Young
expects the program to be completed in about 3
years.
Resources:
Paris: http://www.tds-tds.com/parfact.htm
Douglas Young: 513-569-7624 /
young.douglas@epa.gov
-------
Program for the Identification and
Replacement of Endocrine
Disrupting Chemicals
Douglas Young
US EPA
National Risk Management Research
Laboratory
Cincinnati, OH
-------
Project Outline
©Still in planning phase
© Develop a unique method for identifying potential
EDC (external)
© Develop a program to aid in suggesting
replacements for EDC (internal)
© If possible, combine into one software program
-------
Identifying Potential EDC
© Use a QSAR type approach to quickly identify
potential EDC (external)
© No duplication of concurrent efforts (such as
COREPAandCoMFA)
© Use as another validation tool for concurrent
efforts
©There is a Request for Proposals (RFP) to
establish a collaborative agreement currently
underway
-------
Identifying Potential EDC cont'd
© Collaborator to be largely responsible for this
portion of the project (Thesis or Dissertation
work)
©We can help collect data needed to fill library
-------
Replacement of Potential EDC
© Develop software that will aid in suggesting
replacements for known or potential EDC
(internal)
© Off shoot of the PARIS II Project (Program for
Assisting the Replacement of Industrial
Solvents)
-------
PARIS II
© Suggests possible replacements for currently
used solvents or solvent mixtures
© Uses the DIPPR (Design Institute for Physical
Property Relationships) and UNIFAC to estimate
20 physical and chemical properties of the
solvent(s) to be replaced
© Tries to find best match within database of over
1500 chemicals
-------
PARIS II
cont'd
© If it can't find a single replacement, it will allow
the user to design a mixture
« The user chooses the primary component and then
the program will find the chemical that would form
the best mixture
© If it can't find two component replacement, it will
allow the user to add a 3rd chemical to the
mixture
©Adinfinitum
-------
EDC version of PARIS II
©The framework and coding are established
© Select key physical and chemical properties to
satisfy the requirements for the specific EDC
application
© Might not work with compounds that are
designed to be pesticides & herbicides
-------
Example
© Di(2-ethylhexyl)phthalate (DEHP)
* Used as a plasticizer for PVC medical tubing
"* FDA has released a safety assessment on DEHP
« Found that infants in certain circumstances may be
exposed to unacceptable levels of DEHP
© Replacement outline
* Describe qualities that are required for a plasticizer
« Search the database for possible replacements
-------
Example Cont'd
© Replacement Outline
* Describe qualities that are required for a plasticizer
• Plasticization Efficiency • Solubility
• Tensile strength • Toxicity
• Vapor Pressure • Viscosity
« Match properties using components in database that
have less potential for ED activity
* Suggest a replacement or replacement mixture
-------
Final Product
© A software program that will have a methodology
to identify potential EDC and then will allow the
user to design possible replacement chemicals
(non potential EDC) based on desired properties
© Will be used as another validation tool to
compliment the concurrent research projects
-------
* Determine collaborator during Winter of 2002
* Project will begin Spring/Summer of 2002
« Expected to last 3 years
© Internal
« Post-doc already in place
* Project began January of 2002
« Expected to last 3 years
-------
Region 5 Endocrine Disruptor
(ED) Efforts
Peter Howe (WD)
Lawrence Zintek (CRL)
Dennis Wesolowski (CRL)
Al Alwan (WD)
John Dorkin (WD)
George Azevedo (WD)
Mari Nord (CRL)
Marc Tuchman (GLNPO)
Babu Paruchuri (CRL)
-------
Involvement
Region 3
Jennifer Gundersen
Ron Landy
USGS
John Gannon
Steve Smith
Division Directors
Jodi Traub
Gary Gulizean
Norm Niedergang
Rob Springer
P
Team
oratory
Office of
USDA
Cliff Rice
USGS
Larry Barber
)PPT
Don Rodier
Dave DiFiore
John Walker
ORD
Myriam Medina-Vera
Environment Canada
Donald Bennie
-------
Are These Compounds Present in the
Region at Effect Levels Reported in the
Literature?
[. AlkyIphenols- NP, NP1EO, NP2EO,
NP1EC, NP2EC and Octylphenol
-Degradation Products of Nonionic Surfactants
-Attributed to Fish Endocrine Disruption
I. Sex Hormones (Estrogens/Androgens)
-Natural and Synthetic
-------
Why is Region 5 Involved?
Researchers Have Shown APES to Exist at
Levels of Concern
These Chemicals Cause Endocrine
Disruption in Fish
Needed Analytical Standards
Needed a Standard Method That States and
Environmental Labs Could Use
-------
This Presentation
Alkylphenol Background
Region 5 Central Regional Laboratory Methods
Initiative
Region 5 Water Division Studies
A. Sediment
-Chicago River
-DuPage River
-Des Plaines River
-Fox River
B. Water (New Data)
-Chicago River
-Calumet-Sag Channel
-Lower Des Plaines River
-Illinois River
-------
This Presentation
GLNPO-USDA Ongoing Fish Study
USGS Work in Progress/Future Work
Region 5 Work in Progress/Future Work
Conclusions
-------
Alkylphenol Compounds
How Are They Used ?
Industrial, Institutional and Domestic Surfactants
Antioxidant in Plastics
PVC Stabilizer
Oil Additives
Oil Field Recovery
Metal Extractants
-------
What Happens When
Nonylphenol Ethoxylates Are
j ± j
Released Into The Environment ?
-------
Common Environmental Metabolites of Nonylphenol
Ethoxylates (Adapted from Ahel et al.9 1994; Naylor, 1992)
C9H19
OCH£H7-)-OH NPethoxylate
' n=ltolOO;avg. = 9-17
^\. high aeration
OCH2CH2OCH2CH2OH
C9H19
C9H19
NP2EO
OCH2CH2OH
NP1EO
OCH2CH2OCH2COOH
NP2EC
OCH2COOH
NP1EC
low
C9H19
OH NP
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Bioconcentration Factors (BCF) for
Nonylphenol
Fathead Minnow 271 -984
Three-Spined
Stickleback
Mussel
Mussel
1300
WardandBoeri(1991a)
Brooke (1993)
Ekelundetal.,(1990)
McCleeseetaL, (1980)
EkelundetaL, (1990)
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The Elusive Bottom Line On Endocrine
Disrupting Effects
Fathead Minnows
- Stimulated Production of Female Hormone at 50-
lOOppt
- Levels of NP Greater Than 3.4 ppb Ceased Egg
Production (Giesy et al., 2000)
- NP at 1.6 ppb had an Adverse Effect on Sertoli
Cells (Miles-Richardson et al., 1999)
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The Contribution of Ammonia, Metals and
Nonpolar Organic Compounds to the Toxicity of
Sediment Interstitial Water from an Illinois River
Tributary (Schubauer-Berigan and Ankley, 1991)
TIE conducted on the pore water from Calumet
Sag Channel sediments indicate that
nonylphenols are attributed to the toxicity of the
sediments to Ceridaphnia dubia, a small
Crustacea that lives in the interface of the water
and sediment that is a marker to indicate toxicity
depending on its reproduction and mortality.
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World Regulatory Activities
APE Bans, Phase outs or Use Restrictions
- Denmark
- Japan
- Germany
- United Kingdom
- Belgium
- Switzerland
- Chile
- Sweden
- Netherlands
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Recent Regulatory Activities
Environment Canada Draft WQC = 1 & 0.7
ppb NP for Freshwater and Saltwater,
Respectively
EU Completed Their Risk Assessment and
Recommended Banning "Down the Drain"
Uses
Japan Draft WQC = 0.6 ppb
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Relevant Nonylphenol Concentrations
Concentration fPPB) Effect/Criterion
6 ppb- Fresh Water
1 ppb- Salt Water
0.05-0.10
Draft U.S.WQC
Lowest Effect
Concentration in
Multigenerational Fish
Study
Draft Canada WQC
Stimulation of E2, E^
Production, VTG
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Documented Range of Alkylphenol Concentrations in Large
Region 5 Sewage Treatment Plant Effluents (ppb)
(Larry Barber, USGS Survey)
Compound Chicago Area
NP1EO
NP2EO
NP3EO
NP1EC
NP2EC
NP3EC
NP4EC
1.4-1.7
4.1-13
3.7-7
ND
16-29
44-92
4.1-9.5
1.3-6.1
Minnesota
0.9-2.1
5.8-12
0.78-19
ND-2
21-60
56-100
7.8-13
2.3-6.5
Detroit
Concentrations are Above Effect Level for ED Based on the Literature
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Region 5 CRL Methods Initiative
CRL Director (Dennis Wesolowski)
Developed Methods Initiative Proposal for
40 CFR 136 Incorporation, a Cooperative
Effort with ORD Cincinnati and Region 3
- Completed Sediment SOP for NP, NP1EO,
NP2EO, Octylphenol and Bisphenol A
Analysis by (GC/MS/Full Scan), March 2001
- Completed Water SOP for NP, NP1EO,
NP2EO, OP and Bisphenol A Analysis by
(GC/MS/SIM), September 2001
- Completed Synthesis and Commercial
Availability of Standards with Known Purity
-------
Region 5 Central Regional Laboratory Contribution
Completed Sediment Method
Detection Limits (nnVA
Nonylphenol
NP1EO
NP2EO
Octylphenol
Bisphenol A
110
218
433
Completed Water Method
Detection Limits (nnfk
•Nonylphenol
•NP1EO
•NP2EO
•Octylphenol
•Bisphenol A
61 (Below Effects Criterion Levels)
265
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Commercially Available Standards and Surrogates
Compound
Source: Catalogue Number
n-NP
NP1EO
Aldrich: 29,085-8
Cambridge : ULM-4559
Aldrich: Q2268-5
n-NPlEO
Cambridge: ULM-4520
NP2EO
Aldrich: Q2044-5
n-NP2EO
Cambridge: ULM-4521
NP1EC
Cambridge: ULM-1688
NP2EC
Aldrich: Q2109-3
n-NP2EC
Cambridge: ULM-4690
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Water l)ivisio[n Study
Chicago
Waterways
Sediment Sample
Locations
Red Rivers- Secondary
Contact
North Side WKP
uent Dominated
Stickiiev
North Branch
Diirage River
nt Dominated
Fox River
33% Efflueri
Des Flames Rivei
% Effluent Dominated
Ictke Michigan
Clue ago River
Calumet River
Water R edamati on Plant (WRP)
Sample Locations
C ounty B oundarie s
4 8 Mile
Mul Hold 5 Sept. 2001
-------
North Branch
Sediment Grab Samples Ranges of
Alkylphenols in ppm
Compound
Range in
Octylphenol
Nonylphenol
Nonylphenol Monoethoxylate
Nonylphenol Diethoxylate
0.1-1.2
2.5-48
ND-49
-------
Lower Des Plaines River
Top Strata of Core Samples Range of
Nonylphenol in ppm
Compound
Nonylphenol
Range in
0.32-13.7
-------
DuPage River Composite Sediment
Dam Samples Ranges of Alkylphenols
in ppm
Compound
Octylphenol
Nonylphenol
Range in
ND-0.39
ND-2.5
-------
Fox River Sediment Samples
Many Samples Contain NP and OP but All
Below Reporting Limit.
-------
Chicago Area to Peoria River Water Survey
APES survey, North-side Chicago to Peoria. Winter 2001
-------
Chicago Area to Peoria River Water Study
5000
4000
3000
2000
1000
0
OP
NP
*- NP1EO
*-NP2EO
•*-Bisphenol A
North
South
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GLNPO-USDA
Ongoing Fish Study
Alkylphenols, Alkyl Ethoxylates and Their
Metabolites as Potential Great Lakes,
Tributaries and Effluent Dominated Stream
Endocrine Disruptors. (Cliff Rice, USD A)
-------
North Branch - Chicago River
Fish Collection/Analysis Information
• Collected by Electroshocking at RM 330-
Diversey
• Analyzed for Nonylphenol-APEs (0-5
ethoxy-substituted) by LC/Fluorescence
with Purified Standards
• Confirmation by GC/MS/NCI and LC/MS
-------
Preliminary Results for Total Alkylphenols
(NP, NP1EO and NP2EO) in Carp
North Branch - September 1,1999
(Clifford Rice, USD A, Beltsville, MD)
#31 OR #312
-------
St. Paul Minnesota Metro POTW Walleye
Alkylphenol Concentrations in PPM
(Clifford Rice, USD A, Beltsville, MD)
o.
o.
3
2
1
0
1
i—
NP
NP1EO
NP2EO
NP3EO
6R
FISH ID
Nonylphenol Half Life in Muscle and Fat of Rainbow Trout -19 Hours
(Lewis and Lech, 1996)
-------
GLNPO-USDA Fish Study
(Cliff Rice,USDA)
Lower Des Flames River
- Total NPEs (0-3) 1.55 ppm
Abnormal Left Gill
- Masses on Gonads
North Branch Chicago
River-Fish #312
- Total NPEs (0-3) 33.3 ppm
- Tumerous Growths on
Ovaries , One Large and
One Small
- Large Spleen
-------
Work in Progress
GLNPO-USDA Fish Study
Serum Testosterone, Estrogen and Vitellogenin
Levels and Evaluating Gonad Histopathy for
Fish
Analyzing Fish Tissue for Polybrominated
Diphenol Ethers (Flame Retardants) Which
Appear to be an Emerging Issue
-------
Work in Progress/Future Work
Proposed Study from the USGS
Minneapolis, MN - METRO POTW
Larry Barber and Kathy Lee
Fathead Minnow Study
Toxicity Identification Evaluation (TIE)
- Serum VTG
- Testosterone/Estrogen
- Gonad Histopathy
-------
Hormone Work in Progress
(Larry Barber et al., USGS)
Investigation of the Fate of Natural and
Synthetic Sex Hormones in Sewage
Treatment Plant Effluents
Determine Fate in Receiving Stream
Conduct TIE (Toxicity Identification
Evaluation) to Determine Major
Contributors to Fish ED
-------
Effects of Exposure to 17 (3-Estradiol to
Fathead Minnows
(Miles-Richardson et al., 1999)
• Threshold for Histologic Changes in Testes
-0.04 ppt
• Threshold for Vitellogenin Induction
-0.04 ppt
-------
p-Estradiol Effluent Concentrations at
Four Michigan STPs
(Snyder et al., 1999)
• Average Concentration 1.5 ppt
(Range= ND to 3.7 ppt)
Sewage Treatment Effluent Concentrations 10-30
Fold Higher than LOEC for Endocrine Disruption
Others- Including Ethynyl Estradiol
-------
Region 5 Work in
Progress/Future Work
Improve Techniques/Methodologies
- Solid Phase Extraction
- GC Columns/Conditions/Derivatization
- Other Options
Sediment and Water Monitoring
Investigate Other Compounds of Interest
-NPlECandNP2EC
- Hormones and Pharmaceuticals
-------
Conclusions
NP, NP1EO, NP2EO and OP in Water
Column and Sediment
Need to Include NP1EO and NP2EO with
NP to Assess Total Exposure To Biota
Sediments are Important Reservoirs
Must Work on Exact Chemistry of Toxic
Component and Not Generalize to Mixtures
-------
Conclusions
APEs are Present in the Water Column from Chicago
to Peoria (over 100 miles studied)
- Detected at Lower Concentrations Downstream due to
Dilution, Deposition and/or Decomposition
Standard Methods Enable Others to Sample/Gather
Information to Determine APES Concentrations
Nationwide
Drives Positive Environmental Outcomes
- Enhance Leverage for Voluntary Reduction
- Enables Regulation and Develop Potential Banning
- Enables Setting of Permit Requirements
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