1. Asessment of Endocrine Disrupting Compounds in the Assabet River and its Tributaries Dr. Kathleen Arcaro Kasie Auger April 21, 2011

2. Water was collected from 5 locations in the Assabet River and its tributaries Kasie Auger, Marc Zimmerman, and John Colman collected water from 5 sites in early August of summer 2010 This water was brought back to the lab at Umass Amherst where the exposure studies were conducted

3. Sites Sites were chosen to include a range of impacted environments – Wastewater impacted sites such as Assabet at Rte 9, Westboro – Non-point source or septic influenced sites such as Fort Pond Brook and Coles Brook, Acton

4. Materials Seven tanks included in the exposure 1 negative control containing laboratory water 1 positive control containing 1 ppb E 2 5 tanks each containing the water samples collected from the Assabet River and its tributaries 35 male Japanese Medaka 5 for each exposure

5. Methods 1000 mL of each water sample was placed into its respective tank 5 male Japanese Medaka were placed into each tank At 24 hours an 80% renewal of each tank was performed At 48 hours fish were sacrificed and tissues were harvested

6. Methods RNA was isolated RNA was quantified and diluted to.1μg/μL Real Time RT-PCR measured expression of two genes VTG and L7 in Relative Fluorescent Units With this information relative expression levels were graphed …

7. Results From The First Round Of Testing Increased VTG expression was not observed in male Japanese medaka exposed to Assabet River water Increased VTG expression was not observed in male Japanese medaka exposed to Assabet River water Increased VTG Expression Was Not Observed In Male Japanese Medaka Exposed To Assabet River Water And Its Tributaries