National WWTP effluent screening study
James M. Lazorchak1, Marc Mills2, and Gregory Sayles2
1U.S. Environmental Protection Agency, Office of Research and Development, National Exposure Research Laboratory
2 Environmental Protection Agency, Office of Research and Development, National Risk Management Research Laboratory
Din 2003 and 2004, EPAORD's National Exposure Research Laboratory and National
Risk Management Research Laboratory collaborated on a study of endocrine disruptors in
waste water. Samples were collected of 50 effluents from waste water treatment plants
across 10 EPA Regions. Estrogenic activity in water samples was measured using
EPA/ORD's molecular diagnostic indicator methods. This effort began EPA/ORD's efforts
to study EDCs in the environment at more than a local scale.
Results
Example of Vg Expression in Male Fathead Minnows
Exposed To Wastewater Effluents
Example of NO Vg Expression in Male Fathead Minnows
Exposed To Wastewater Effluents
Research Goals
Table 1. Estrogens and Androgens in the *12 Positive Effluents (Prelim 2004)
Reports of potential wildlife risk from exposure to environmental estrogens emphasize the
need to better understand both estrogenic and androgenic presence and persistence in
treated wastewater effluents. In addition to wildlife exposure, human exposure should also
be examined, especially in situations where estrogenic effluents may return to a drinking
water supply. This potential has been examined in rivers and reservoirs in the U.K. where
they found reduced estrogenicity downstream from wastewater outfalls and no
estrogenicity in reservoirs receiving these waters.
In 2000 NERL and the University of North Texas collaborated in a study where the toxicity
and estrogenicity of a final treated municipal effluent was examined. Male fathead
minnows were deployed in the effluent for 3 weeks. Vitellogenin (VTG) protein, Gonado-
Somatic Index (GSI), Hepato-Somatic Index (HSI) and secondary sexual characteristics
were biomarkers used in fish models to assess aqueous estrogenicity. Vitellogenin gene
expression was also measured. There was a very good correspondence in the biomarker
results and the gene expression results. This study provided evidence that the vitellogenin
gene expression assay could be used to assess estrogenic EDC exposures in effluents.
In 2002 and 2003 NERL and NRMRL contacted our 10 Regional Biologists, Regional
Science Liaisons, EPA inspectors, State EPA inspectors and Municipal plant operators to
voluntarily collect up to 50 effluents and ship them to Cincinnati for male and female
fathead minnow exposures. The objective of this study was to determine if adult male or
female fathead minnows exposed to a municipal waste water treatment plant (WWTP)
effluent elicit a change in vitellogenin gene expression above or below a lab water control.
The goal was to use vitellogenin gene expression results to obtain a rough idea of how
common estrogenic and androgenic effluents are in the U.S.
Effluent Sample Source
WA4B
NM2
CT3
MT1
WV1
GA1
MA1
WA4A
TX4
VA3
CT1
WA1
Progesterone
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