Asessment of Endocrine Disrupting Compounds in the Assabet River and its Tributaries

Dr. Kathleen ArcaroKasie Auger

April 21, 2011

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

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

Materials• Seven tanks included in the exposure• 1 negative control containing laboratory water• 1 positive control containing 1 ppb E2

• 5 tanks each containing the water samples collected from the Assabet River and its tributaries

• 35 male Japanese Medaka 5 for each exposure

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

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 …

Results From The First Round Of TestingIncreased VTG Expression Was Not Observed In Male Japanese Medaka

Exposed To Assabet River Water And Its Tributaries