The Ecological Effects of Endocrine Disruption Dr. David Walker University of Arizona

1. The Ecological Effects of Endocrine DisruptionDr. David WalkerUniversity of Arizona David Walker1,Nick Paretti2, Gail Cordy2, Timothy S. Gross3, Edward T. Furlong4, Dana W. Kolpin5, and Dennis McIntosh6 1University of Arizona, Environmental Research Laboratory, 2601 E. Airport Dr., Tucson, AZ 85706 dwalker@ag.arizona.edu 2USGS., WRD, 520 N. Park Ave, Suite 221, Tucson, AZ 85719 nvp@email.arizona.edu 3 USGS-Florida Caribbean Science Center, 7920 NW 71st St., Gainesville Florida, 32653 tim_s_gross@usgs.gov 4 USGS, National Water Quality Laboratory, Denver Federal Center, P.O. Box 25046, MS 407, Lakewood, CO 80225-0046 efurlong@usgs.gov 5 USGS, WRD, P.O. Box 1230, Iowa City, IA 52240 dwkolpin@usgs.gov 6 Delaware State University, 1200 N. DuPont Highway Dover, DE 19901 dmcintosh@desu.edu

2. For Our Purposes… • An endocrine disruptor is a synthetic (anthropogenic) chemical that when absorbed into the body mimics, blocks, or otherwise alters hormone level, function, or binding and subsequently disrupts normal bodily functions including behavioral and/or strictly physiologic responses.

4. Aquatic Ecology and Endocrine Disruption • An individual organisms ability to better-exploit a resource (or group of resources) in the face of environmental stress and inter-specific competition, coupled with conservation of the genetic material enabling this exploitation, is what drives speciation.

5. Genetic conservation of traits is initiated, and sustained by, subtle behavioral cues for mating, spawning, aggression, territoriality, avoidance, etc. • Any impairment of these behavioral cues or manifestation into physiological or morphological changes has the capability to stunt speciation by lowering fertility and fecundity.

6. Endocrine Disrupting Compounds • FAR more than what can be included in this presentation. • By the time breakdown products and metabolites are added to the mix, iterations become astronomical.

7. Just a Few Examples by Use Category • Detergent Metabolites • Fire/Flame Retardants • Fragrances/Flavors • Fuels/PAH’s • Herbicides/Insecticides • Household Wastewater Compounds • Non-Prescription Drugs • Plasticizers/Antioxidants • Prescription Drugs • Steroids

8. At the landscape scale, those compounds known to be “powerful” EDC’s, but are not environmentally-persistent, exert less of an effect than those persistent, but relatively weaker, compounds.

9. Quantification and Research Design Issues; One Size Does Not Fit All Genetic Mechanistic Understanding Biochemical Physiological Histopathological Behavioral Immunological Reproductive Bioenergetic Assemblages Populations Ecological Significance

10. Observational versus Controlled Studies • True control and replication is not possible in the field. • Laboratory studies with control and replication give up some ecological significance.

11. Exposure and Causation • Several studies have examined the effect of one or a very few EDC’s on the physiological response of an organism. • The vast magnitude of compounds in a matrix makes assumptions about individual compounds difficult to ascertain. • Non-monotonic dose-response curves

12. With new and emerging contaminants found almost on a daily basis, making assumptions about exposure and physiologic response must always carry the caveat “of the compounds we analyzed for”…

13. Grab, Composite, or Integrated Samples? • Problems associated with not knowing long-term exposure to fish or other organisms.

14. Passive Organic Chemical Integrative Sampler (POCIS) and/or Semi-Permeable Membrane Devices (SPMD’s aka “fatbags”)

15. Of all the tools at our disposal to study complex environmental issues in aquatic ecosystems, a sound understanding of ecological principles as they pertain to these ecosystems is the most essential.

16. Quantifying Endocrine Disruption in a Threatened and Endangered Fish Species

18. Unlike semi-arid or north-temperate regions, effluent-dependent water’s (EDW’s) in arid regions usually contain 100% effluent year-round.

19. The Santa Cruz River Near Tucson, Arizona • Flows from Mexico near Nogales, Sonora northward to Tucson, Arizona. • The only sections with flowing water are those due to discharge from WWTP’s.

21. Roger Road WWTP • Built in 1951. • Treated effluent is discharged into the Santa Cruz River or diverted into the city’s reclaimed water system. • Treats the wastewater generated by a population of about 419,000. • A capacity of 41 mgd and treated an average of 38 mgd from 2004 to 2005. • Produces secondarily-treated wastewater

22. Tucson Santa Cruz River Roger Road WWTP

24. This Study • Laboratory study with controls, replicates, and randomization. • Use fish native to the region (largely pollution-tolerant). • Framework or foundation for refinement of future studies. • Varying doses of effluent (“treatments”). • Concentrate on long-term, persistent compounds.

25. Bonytail Chub (Gila elegans)

26. Control Control Treatment Treatment

27. Water temperature maintained between 25-29o C. Photoperiod was maintained at 12 hours of light and dark

30. Treatment/Dosages • Fish in raceways exposed for 3 months per treatment • 1st treatment = 1/3 by volume treated ww and 2/3 water treated by RO • 2nd treatment = 2/3 by volume treated ww and 1/3 water treated by RO • 3rd treatment = full strength treated ww

31. Results

34. Males - Overall 17β-Estradiol Control (n = 6): 217.3 Treatment (n = 13):= 547.4 11-ketotestosterone Control (n = 6): = 820.8 Treatment (n = 13):=473.5 Vitellogenin Control (n = 6): = 0.09 Treatment (n = 13):= 0.32

35. Females - Overall 11-ketotestosterone Control (n = 54): 591.3 Treatment (n = 47):530.4 17β-Estradiol Control (n = 54): 568.2 Treatment (n = 47): 403.7 Vitellogenin Control (n = 54): 0.18 Treatment (n = 47): 0.18

38. Synergistic Effects • Ratios of primary male and female sex hormones, in “undisturbed” populations would be expected to have an inverse relationship i.e. as one increased, the other would decrease. • We could therefore assume that major deviations from this inverse relationship between male and female primary sex hormones, could be attributed to impairment.

39. Control Males Treatment Males

40. Control Females Treatment Females

41. Synergism, feedback mechanisms, and non-linearity of bio-markers makes data reduction necessary to determine trends. • Ordination is a good statistical tool but still assumes some degree of linear correlation as would occur with a typical dose-response curve.

42. Males

43. Females

44. Summary • Significant hormonal impairment of both sexes, as compared to controls, at very low concentration of compounds. • This impairment could never have been determined in a field study. • Commonly-used parametric analyses are often inadequate in determining impairment.

45. Summary (cont) • Determination of either hormonal impairment or endocrine disruption requires using phased biomarkers. • Phase 1: Aromatase/GnRH • Phase 2: GtH I, GtH II • Phase 3: Sex hormones • Phase 4: Protein development (vtg, oocyte, spermiation) • Phase 5: Intersex

46. Current and Future Research • Fertility/fecundity and sex ratio/development of F2 generation. • Behavior. • Treatments using streambed sediment from affected EDW’s.

47. This study is highly representative of the biological effect of endocrine-disrupting compounds at the landscape scale.

48. Questions?