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Project Title: Lake Endocrine Disruption
Project Number: GC11NQ00DRC0000 - 8607DRC - MN 257
Project Chief: Kiesling, Richard L.
Project Start Date: July, 2009
Project End Date: June 30, 2013

The study assesses vulnerability of lake-fish communities to endocrine disruption from water and sediment using pond mesocosms. Adult fathead minnows are introduced into mesocosm tanks and allowed to spawn. Experimental exposures to 17-beta estradiol are conducted for adults and fry. Physical and chemical are collected weekly. Water samples from the mesocosm tanks and the feedstock tanks are collected and analyzed for 17-beta estradiol. Significant differences are maintained between controls and treatment doses. Physical and chemical data are collected from the mesocosm treatment tanks.

Statement of Problem:
Recent work has identified on-site septic systems as a source of emerging contaminants to lakes (Carrara and others, 2008; Godfrey and others, 2007), and recent results from whole-lake experimental exposures to endocrine disruptors have documented complete reproductive failure of short-lived species of fish (Kidd and others 2007), raising questions about the effects of these compounds on longer-lived species of fish common to Minnesota lakes. Preliminary data from ongoing research in Minnesota have found biomarkers of endocrine disruption in female fish from urban lakes (Schoenfuss and others, unpublished data) raising questions regarding how Minnesota fish communities will respond to EAC exposure. Given the large number of lakes at risk from point and nonpoint sources of emerging contaminants, there is a need for relevant data on how vulnerable fish populations respond to chronic EAC exposure from contaminated lake water and sediment.

Objectives:
Recent work has identified on-site septic systems as a source of emerging contaminants to lakes (Carrara and others, 2008; Godfrey and others, 2007), and recent results from whole-lake experimental exposures to endocrine disruptors have documented complete reproductive failure of short-lived species of fish (Kidd and others 2007), raising questions about the effects of these compounds on longer-lived species of fish common to Minnesota lakes. Preliminary data from a statewide survey of endocrine disruption in Minnesota lakes have found biomarkers of endocrine disruption in fathead minnows and bluegills fish from urban storm-water lakes and rural septic-system lakes (Schoenfuss and others, unpublished data) raising questions about how fish populations in Minnesota lakes will respond to EAC exposure. Given the large number of lakes in Minnesota at risk from septic tank and nonpoint sources of emerging contaminants, there is a need for relevant data on how vulnerable fish populations will respond to EAC exposure from contaminated lake water and sediment during development and reproduction.

Our project will assess the vulnerability of lake fish communities to endocrine disruption using pond mesocosms located at the USGS Upper Mississippi Ecosystem Science Center (UMESC). Our first study objective is to assess developmental impacts of juvenile exposure to water and sediment concentrations of EACs. Our second objective is to assess reproductive impacts of juvenile and adult exposure to water and sediment concentrations of EACs.

Strategy and Approach:
1: Developmental impacts of juvenile exposure to water and sediment concentrations of EACs.

Effects of EACs will be measured in a three-way, balanced design using four-cubic meter mesocosms. Mesocosms will be installed in 0.1 acre ponds located at the USGS Upper Mississippi Ecosystem Science Center (UMESC). Three species of juvenile fish reared at the UMESC facility (fathead minnow Pimephales promelas, bluegill (Lepomis macrochirus), and walleye (Stizostedion vitreum) or yellow perch (Perca flavescens)) will each be exposed to one of three EACs (ethynylestradiol, 4-nonylphenol (4-NP), or nonylphenol ethoxylate (NPEO)) through either water or sediment exposure for six weeks. Water and sediment concentrations will be monitored four times during the experiment, and fish will be evaluated using blood-chemistry biomarkers (e.g., plasma vitellogenin) and histo-pathological indices of abnormal development (e.g., intersex). Half of the fathead minnows and bluegills exposed in this experiment will be allowed to grow to sexual maturity and will be evaluated for reproductive success as part of Result 2 below.

2: Reproductive impacts of juvenile and adult exposure to water and sediment concentrations of EACs.

Effects of EACs on reproductive success (i.e, mating behavior and spawning success) will be measured in a two-way, balanced design using four-cubic meter mesocosms. Mesocosms will be installed as above in 0.1 acre ponds. Juvenile fathead minnows and bluegills reared under Result 1 above will be allowed to spawn under controlled conditions and evaluated for reproductive output. Sexually mature adults of walleye or perch from common stock populations at UMESC (and bluegill if necessary) will each be exposed to ethynylestradiol through water or sediment exposure for six weeks prior to spawning. Water and sediment concentrations will be monitored four times during the experiment, and fish will be evaluated using blood-chemistry biomarkers (e.g., plasma vitellogenin) and histo-pathological indices of abnormal reproductive physiology before and after spawning as appropriate.

Progress during FY10:
Mesocosm were constructed and tested. Adult fathead minnows were introduced into the mesocosm tanks and allowed to spawn. The first experimental exposures to 17-beta estradiol started for adults and for fry. Physical and chemical data were collected weekly during the experiments. Water samples from the mesocosm tanks and the feedstock tanks were collected weekly and analyzed for 17-beta estradiol using ELISA methods. Significant differences were maintained between controls and treatment doses through the 6 week experiment. Physical and chemical data were also collected weekly from the mesocosm treatment tanks. Fish survivorship was high in both adult and fry treatments, providing sufficient stocks for over-wintering of fry.

Plans for FY11:
Build a population database for samples analyzed using ELISA methods at USGS lab. Provide LCCMR with project plan status report, and maintainfathead juveniles in culture through the winter.