State Water Resources Research Institute Program

Project ID: 2007MA74B
Title: Development of a standardized protocol for fish bioassays detecting estrogenic exposure
Project Type: Research
Start Date: 3/01/2007
End Date: 2/29/2008
Congressional District: First
Focus Categories: Methods, Non Point Pollution, Wastewater
Keywords: medaka, vitellogenin, estrogens, bioassay, real time RT-PCR
Principal Investigators: Arcaro, Kathleen Francis; Moffatt, Lauren Taylor
Federal Funds: $ 4,917
Non-Federal Matching Funds: $ 37,984
Abstract: Endocrine disrupting compounds are of increasing concern in waterways throughout the Northeast. Specifically, estrogenic contaminants have been detected downstream of agricultural operations, industrial discharges, and even municipal wastewater treatment facilities. Concern regarding the feminization of fish and wildlife species has led to the demand for sensitive, accurate, and reliable means to detect estrogenic activity in water samples. Fish models are ideal bioassays for examining potentially polluted waterways because fish are directly exposed to, and quickly concentrate aquatic contaminants. In concert with the use of molecular techniques, specifically the analysis of changes in gene expression, fish bioassays can be a sensitive indicator of estrogenic pollution. However, studies using fish bioassays have reported variable results with respect to assay sensitivity and robustness of response. Part of the variability among results is due to differences in experimental setup, exposure conditions, and endpoints measured. I propose to optimize the protocol for detecting estrogenic compounds in water samples using male Japanese medaka (Oryzias latipes). Male medaka will be exposed to the endogenous hormone 17Β-Estradiol and assay parameters including length of exposure, number of fish per tank, volume of water per tank and number of water changes will be systematically examined. I will then use quantitative real-time RT-PCR to quantify changes in a panel of estrogen-responsive genes in each of the experimental groups. Results form this study will help establish a standard operating procedure for detecting estrogenic compounds using medaka, and provide a guideline for optimizing other fish bioassays.

Progress/Completion Report, PDF

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