2011 Progress Report: Transport/Fate/Ecological Effects of Steroids from Poultry Litter & Evaluations of Existing/Novel Management Strategies

EPA Grant Number: R833418
Title: Transport/Fate/Ecological Effects of Steroids from Poultry Litter & Evaluations of Existing/Novel Management Strategies
Investigators: Fisher, Daniel J. , Kane, Andrew S. , Klauda, Ronald J. , Staver, Kenneth , VanVeld, Peter , Yonkos, Lance T.
Institution: Wye Research and Education Center , Maryland Department of Natural Resources , School of Medicine at the University of Maryland , Virginia Institute of Marine Science
EPA Project Officer: McOliver, Cynthia
Project Period: August 1, 2007 through June 30, 2010 (Extended to July 31, 2012)
Project Period Covered by this Report: January 1, 2011 through December 31,2011
Project Amount: $694,736
RFA: Fate and Effects of Hormones in Waste from Concentrated Animal Feeding Operations (CAFOS) (2006) RFA Text |  Recipients Lists
Research Category: Endocrine Disruptors , Health , Safer Chemicals


The proposed research addresses all three areas of interest contained in the EPA STAR grant RFA:  fate of poultry litter-associated hormones, impacts of exposure on ecological systems, and evaluation of various management strategies. Given that fecal sex steroids persist at high concentrations in poultry litter, we hypothesize that runoff from fields treated with litter will exert a direct steroidal effect on aquatic organisms within receiving waters. Previous studies at our laboratory have demonstrated that:  (1) fecal steroids in poultry litter reach receiving waters via rain-induced runoff; (2) this runoff is sufficiently estrogenic to feminize male fish; and (3) differences in agricultural management strategies can affect steroid concentrations in runoff and receiving waters. The proposed research will address remaining questions concerning environmental persistence and bioactivity of steroids upon reaching surface waters and further investigate effects of agronomic practices on mitigating resultant environmental steroid loads.

Progress Summary:

  1. Determine the efficacy of a Subsurface Litter Injector in reducing/eliminating the runoff of steroids from litter amended watersheds.

    Subsurface injection of poultry litter resulted in an 82% reduction in steroid concentrations in runoff compared to runoff from a No-till litter application.

  2. Determine the frequency and severity of testicular oocytes in male largemouth bass (Micropterus salmoides) from rivers and lakes on the Delaware and Maryland portions of the Delmarva Peninsula.

    Occurrence of testicular oocytes in male largemouth bass (Micropterus salmoides) ranged from 33% to 88% in lakes and rivers on the Delmarva Peninsula. Prevalence of intersex in Delmarva largemouth bass is similar to reported levels of intersex in largemouth bass and smallmouth bass from the Western Shore of Maryland and nationally. Site average Intersex Severity Indices for largemouth bass ranged from 0.11 to 0.46. These values are generally lower than those reported in smallmouth bass collected from the Potomac River system, but higher than those reported for smallmouth bass from minimally impacted reference sites outside of the Potomac Drainage. These are the first reported findings of intersex in waters of the Delmarva Peninsula.

  3. Conduct laboratory experiments exposing adult fathead minnows (Pimephales promelas) to raw poultry litters and a pelletized poultry litter. The exposures were designed to compare estrogenicity changes from aqueous poultry litter solutions over time using both chemical analytical methods and biological methods.

    Aqueous poultry litter mixtures contain abundant conjugated estrogens, which appear to readily deconjugate to free bio-active forms. This results in increases, both in measured estrogens and, more importantly, in estrogenicity as measured by in vitro and in vivo bioassays. Therefore, laboratory assays with continuous flow or daily treatment renewal may underestimate the latent estrogenicity of a given poultry litter dose. Likewise, collection and preservation of runoff at the time of discharge (i.e., T0) may underestimate impacts to receiving waters. Similarly, estrone (E1 - a weak estrogen) converts to 17β-estradiol (E2 - a potent estrogen) when placed in solution. In this way, estrogenicity can increase with time even as total measured estrogens decrease. Estrogenicity, as indicated by in vitro assays, is explained almost exclusively by measured E2 concentration with minimal contribution from E1. Investigators using in vitro and/or in vivo assays often indicate E1 bioactivity to be 10 to 100 fold less than E2. In many instances, this may actually reflect conversion of E1 to E2 in solution. In vitro assays are sensitive indicators of estrogenicity with detection limits often below those of analytical methods. Likewise, induction of Vtg mRNA and resulting Vtg protein are sensitive measures of estrogenicity and can yield indications of exposure to estrogens at or near analytical limits of quantitation.

Future Activities:

All activities proposed in the original grant have been completed.

Journal Articles on this Report : 1 Displayed | Download in RIS Format

Other project views: All 9 publications 1 publications in selected types All 1 journal articles
Type Citation Project Document Sources
Journal Article Yonkos LT, Friedel EA, Fisher DJ. Intersex (testicular oocytes) in largemouth bass (Micropterus salmoides) on the Delmarva Peninsula, USA. Environmental Toxicology and Chemistry 2014;33(5):1163-1169. R833418 (2011)
R833418 (Final)
  • Abstract from PubMed
  • Abstract: Wiley-Abstract
  • Supplemental Keywords:

    endocrine disruption, indicators, vitellogenesis, intersex, testicular oocytes, estrogenicity, environmental exposure, risk assessment, sensitive populations, water, fish, amphibians,  largemouth bass, Micropterus salmoides, fathead minnow, Pimephales promelas, population, ecosystem protection, environmental chemistry, histology, Chesapeake Bay, agriculture, concentrated animal feeding operation, CAFO 

    Progress and Final Reports:

    Original Abstract
  • 2008 Progress Report
  • 2009 Progress Report
  • 2010 Progress Report
  • Final Report