Development of a Bioassay for AhR-mediated Toxicity to Rainbow Trout

EPA Grant Number: R825371
Title: Development of a Bioassay for AhR-mediated Toxicity to Rainbow Trout
Investigators: Giesy, John P.
Current Investigators: Giesy, John P. , Richter, Catherine A. , Blankenship, Alan L. , Villeneuve, Daniel L.
Institution: Michigan State University
EPA Project Officer: Hahn, Intaek
Project Period: December 15, 1996 through December 14, 1998
Project Amount: $304,771
RFA: Exploratory Research - Environmental Biology (1996) RFA Text |  Recipients Lists
Research Category: Biology/Life Sciences , Aquatic Ecosystems


Fish are particularly vulnerable to toxins which act through the aromatic hydrocarbon receptor (AhR). These toxins include dioxins, polychlorinated biphenyls (PCBs), and other halogenated diaromatic hydrocarbons (HDHs). Pulp mill effluents and lampricide formulations have also been found to contain components which activate the AhR. These toxins cause a variety of adverse effects, including immune system dysfunction and reproductive impairment. Since they occur in complex mixtures, the overall exposure to fish in a particular environment is difficult to assess. The problems of chemical analysis and evaluation of potency of the many compounds which bind AhR can be avoided by using a bioassay. The studies are developing and characterizing an in vitro bioassay based on cell lines derived from rainbow trout. The cells are being stably transfected with a plasmid containing the gene encoding firefly luciferase under the control of DREs. The cell line will express luciferase as an integrative measure of the total potency of AhR-active toxins to which it is exposed. The bioassay will be calibrated by comparing it to toxicity tests in whole fish using mortality, biochemical markers, and reproductive impairment as endpoints. The bioassay will be compared to measures of AhR activation and native gene induction. Current tests use native induced genes such as P4501A1, and mammalian cell lines. The proposed bioassay will have the advantages of being directly relevant to fish, and using a gene controlled only by AhR, not by other cellular regulation. The bioassay can be used to predict the toxicity of complex mixtures, investigate interactions among HDHs, and detect sources of AhR-active toxins. The bioassay can be used as a screening test for new chemicals or for use in conjunction with solid phase extraction of residues from water and instrumental analyses in a Toxicant Identification and Evaluation (TIE) protocol.

Publications and Presentations:

Publications have been submitted on this project: View all 8 publications for this project

Journal Articles:

Journal Articles have been submitted on this project: View all 3 journal articles for this project

Supplemental Keywords:

water, p450, biochemistry, transfection., RFA, Scientific Discipline, Toxics, Ecosystem Protection/Environmental Exposure & Risk, Ecology, Ecosystem/Assessment/Indicators, Ecosystem Protection, exploratory research environmental biology, Chemical Mixtures - Environmental Exposure & Risk, Environmental Chemistry, pesticides, Chemistry, Ecological Effects - Environmental Exposure & Risk, Ecological Effects - Human Health, Biology, Ecological Indicators, bioindicator, complex mixtures, dioxin, chemical contaminants, pulp mill effluents, PCB, polychlorinated biphenyls, bioassay, biochemical measurements, hydrocarbons, fish , immune systems, luciferase, reproductive health, dioxin exposure

Progress and Final Reports:

  • 1997 Progress Report
  • Final Report