Intersex and other Indicators of Endocrine Disruption throughout the Northeast

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1 Intersex and other Indicators of Endocrine Disruption throughout the Northeast Luke R. Iwanowicz, Vicki S. Blazer, Kelly L. Smalling USGS, Leetown Science Center, National Fish Health Research Laboratory USGS, New Jersey Water Science Center April 26, 2017

2 What is Intersex? An abnormal condition characterized by the presence of male and female reproductive components in a gonad that should be exclusively male OR female.

3 So what is Normal?

4 FEMALE MALE

5 Intersex Male (testicular oocytes) Immature oocytes Mature sperm

6 Emerging Contaminants (Chemicals of Emerging Concern; CECs) Synthetic or natural compounds that are not commonly monitored or regulated, but are present in the environment and lead to adverse effects Chemically engineered to have a biological effect at low concentrations and may inadvertently effect non-target organisms sub ppb concentrations Contains the group of chemicals called endocrine disruptors

7 Estrogens in the Environment Sewage treatment facilities Confined (intensive) animal production Run-off Pesticides and other emerging contaminants Phytoestrogens Many others..

8 2003 Potomac Watershed: Cause for Concern? Multiple bacterial pathogens Parasite/fungal infections Skin lesions Largemouth Bass virus High prevalence of intersex Immunosuppression Causes?

9 Associations of Land-use with Intersex Spawning Study (Chesapeake Bay Drainage) Site Gauley River South Branch Petersburg South Branch Moorefield South Branch Springfield Shenandoah North Fork Shenandoah Mainstem Shenandoah South Fork Conococheague Creek (lower) Human Density WWTP WWTP Flow % Ag AFO Animal Numbers Intersex % 0.02 (0.07) ,450, % (296) 0.97 (0.95) ,384, % (496) 0.50 (0.50) ,719, % (562) 1.02 (0.76) ,174 11,757, % (960) 1.16 (0.78) ,655 33,928, % (2,539) 1.64 (0.93) ,029 14,788, % (1,176) 1.83 (0.65) ,819, % (1) 1.03 (0.78) Land-use Characteristics Intersex Prevalence Intersex Severity Blazer et al (2012) Reproductive endocrine disruption in smallmouth bass (Micropterus dolomieu) in the Potomac River basin: spatial and temporal comparisons of biological effects. Environmental Monitoring and Assessment, 184: r 2 p r 2 p Human population density Number of WWTP WWTP Flow Percent agriculture Tot. Animal Feeding Operations Poultry houses Total animals Animal density

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11 To understand the population level effects of endocrine disruption resulting from exposure to contaminants in the Chesapeake Bay Watershed Identify and quantify the sources, fate, transport, distribution, and exposure of EDCs to fish and wildlife. Evaluate the effects EDCs on fish and wildlife in the environment. Determine the mechanism(s) and thresholds for adverse effects on fish and wildlife species. Develop appropriate assessment tools and models for risk evaluation of EDCs on natural resources.

12 Northeast Region FWS NWRs Site selection 19 Refuges Assabet River, Back Bay, Blackwater, Cherry Valley, Erie, Great Bay, Great Meadows, Great Swamp, John Heinz, Mason Neck, Missisquoi, Montezuma, Moosehorn, Ohio River Islands, Patuxent, Rappahannock, Sunkhaze, Umbagog, and Wallkill Iwanowicz et al Ecotox Environ Safety. 124:

13 Field Sampling Target species Smallmouth bass (10 per sex) Largemouth bass (10 per sex) Sample dates Sampling occurred during Late August thru late October 2008 Rivers 2009 Rivers 2010 Impoundments Fish collection Pulsed DC electroshocking Angling when necessary

14 Sample collection Morphometric data collected External health assessment Blood collected for plasma Vitellogenin Internal organs removed and preserved in Z-Fix Histopathology Intersex Field Sample Processing Liver and anterior kidney collected for possible gene transcription work Water samples for in vitro bioassays

15 Sample analysis Histopathology - Intersex (testicular oocytes in males) ELISA - Plasma vitellogenin In vitro bioassays nuclear receptor activation Estrogenicity, androgenicity & glucocorticoid receptor translocation

16 Smallmouth bass intersex prevalence Intersex ranged from % (composite = 85%) 118 males evaluated in total No differences between up and downstream locations Previous work has identified intersex as low as 10-14% OH/WV ME NJ VT ME PA/NJ NH/ME

17 Estrogenicity Results Estrogenicity measured at a number of locations Highest concentrations measured in WWTP effluent (over 5ng/L) PNOEL = 0.73ng/L 36% of 38 non-wwtp samples were estrogenic 79% of those were above the PNOEL

18 General Comments Evidence of estrogenic endocrine disruption in waters of FWS Northeast Region NWRs Apparent differences between SMB and LMB SMB intersex 85% (Hinck et al 2009: 33%) (Blazer et al > 90%) LMB intersex 27% (Hinck et al 2009: 18%) The current study did not comprehensively evaluated water quality or site specific emerging contaminants Further research may be warranted to identify the sources and causes (chemical?) biological effects

19 The Full Cast (Co-authors NWR Study) L.R. Iwanowicz, V.S. Blazer, A.E. Pinkney, C.P. Guy, A.M. Major, K. Munney, S. Mierzykowski, S. Lingenfelser, A. Secord, K. Patnode, T.J. Kubiak, C. Stern, C.M. Hahn, D.D. Iwanowicz, H.L. Walsh, and A. Sperry

20 ED in New Jersey Sites selected based on previous monitoring data (WQ and/or bio) and land-use 101 sites sampled in fall under base-flow 16 reservoirs 85 river sites Estrogenicity only in water

21 THANK YOU! FOR MORE INFORMATION Funding provided by US Fish and Wildlife Service Division of Environmental Quality, USGS Priority Ecosystems, Contaminant Biology, Ecosystems and Cooperative Water Programs, NJ Department of Environmental Protection.