Grantee Research Project Results
2008 Progress Report: Fate of Hormones in Tile-Drained Fields and Impact to Aquatic Organisms Under Different Animal Waste Land-Application Practices
EPA Grant Number: R833417Title: Fate of Hormones in Tile-Drained Fields and Impact to Aquatic Organisms Under Different Animal Waste Land-Application Practices
Investigators: Lee, Linda S. , Turco, Ronald F. , Sepulveda, Marisol S. , Jafvert, Chad T.
Institution: Purdue University
EPA Project Officer: Aja, Hayley
Project Period: April 16, 2007 through April 15, 2011
Project Period Covered by this Report: April 16, 2007 through April 15,2008
Project Amount: $700,000
RFA: Fate and Effects of Hormones in Waste from Concentrated Animal Feeding Operations (CAFOS) (2006) RFA Text | Recipients Lists
Research Category: Endocrine Disruptors , Human Health , Safer Chemicals
Objective:
(1) Assess the relative amount of hormones discharged from tile-drained agricultural fields under different manure and lagoon effluent applications; (2) assess hormone persistence in fields under these application practices; and (3) evaluate the impacts of these hormone loads (relevant levels and mixtures) on aquatic organisms.
Progress Summary:
Animal Science Research and Education Facility (ASREC) Site:
During the first year of the project, we established four tile drain and three stream/ditch sampling and flow monitoring stations at the ASREC site. We began water sample collection at each station that was in full operation. Samples are being processed and analyzed for 17α- and 17β-estradiol and trenbolone, and related metabolites (estrone, estriol, trendione, androstendione) using LC/MS. A subset of samples from both drains and streams also are being analyzed for pH, EC, nitrates, phosphate, and dissolved organic carbon; and a subset of stream samples are being analyzed for pesticides used on the associated fields.
Water Quality Field Station (WQFS) Site:
We initiated flow-weighted sampling of drainage from eight 520 m2 plots fertilized with subsurface swine manure injection (4 in the spring prior to planting and 4 in the fall after harvest) at our already established WQFS. 2007 data from this site showed the presence of primarily estrogens in discharge water in response to rain events that occurred approximately three weeks after subsurface injection of swine manure, but all concentrations were lower than concentrations known to invoke negative responses in aquatic species.
Hormone Persistence in Manure-Applied Fields:
We attempted to quantify persistence of hormones in manure-applied fields by subsampling fields subsurface injected with swine manure; however, we encountered several problems: extreme variability between subsamples and challenges with limits of quantitation (LOQ) in the samples during the first 2 weeks after application. LOQs were 3 orders of magnitude higher for water collected from the tile drains such that we may see no quantifiable limits in the field soil-manure subsamples, but continue to see substantial hormone concentrations in the aqueous discharge. Therefore, we chose to focus on assessing persistence in laboratory microcosm studies under a range of temperature and moisture conditions that would represent field conditions. For the hormones of interest, no literature was available on persistence of metabolites excreted from 17β-trenbolone acetate (TBA)-implanted cattle. TBA is converted metabolically to primarily 17α-trenbolone and trendione, and excreted in manure from implanted cattle. To predict the persistence of TBA metabolites once land-applied via manure, aerobic degradation rates in two contrasting agricultural soil types (clay loam and a sandy soil) of both trenbolone isomers (17α and 17β) and their primary metabolite trendione were measured and isomer interconversion was assessed. The impact of manure application also was evaluated in the clay loam soil. At 25°C and optimal moisture conditions, both isomers degraded to trendione in a similar manner with half lives (t½) in the order of a few hours to 0.5 d at applied concentrations of 1 mg/kg. Similar degradation rates also were observed in the presence and absence of manure applied at rates typical for land-application of cattle manure. Trenbolone degradation was concentration-dependent with degradation rates decreasing with increasing applied concentrations. Trendione, whether applied directly or produced from trenbolone, persisted longer than trenbolone with t½ values of 1 to 4 d. A small amount (1.5%) of conversion of trendione back to 17β-trenbolone was observed during aerobic incubation regardless of the applied concentration. A small amount of 17α–isomer also converted back to 17β-trenbolone, presumably through trendione. In autoclaved soils, degradation was small to negligible compared to a microbially active soil.
Sorption Studies:
To aid in predicting transport of manure-borne TBA metabolites, multi-concentration sorption isotherms for 17α- and 17β-trenbolone and trendione were generated with five autoclaved-sterilized soils that represented a range in soil properties. Hormone concentrations were measured independently in solution and soil phases, and quantified using LC/MS.
Hormone Impact on Aquatic Organisms:
We have sampled fish and turtles from streams and ponds receiving runoff from fields treated with animal manure. In order to determine exposure to estrogens in turtles, we have developed methods for the semi-quantification of VTG in plasma of common snapping turtles (Chelydra serpentina). Turtles have been collected during the reproductive season (May-June) in 2007. VTG was detected in plasma samples from all females but absent from male turtles from both reference and CAFO sites. Comparison of the relative abundance of VTG plasma concentration showed that the average VTG concentrations were similar between the two female populations, but that relative VTG concentrations were more variable in the population from the CAFO site than the population from the reference site. In spring 2008 we began exposing fathead minnows (from day 0 to day 30 post-fertilization) to water from Marshall Ditch in a flow-through tank system built inside a small shed. We had mixed results in that our survival was low.
Future Activities:
We will be setting up a wireless communication system at all of the ASREC stations and initiate development of a recession model that would allow us to optimize sample collection during the rise and fall of each hydrograph, which would reduce our overall analysis and labor costs without compromising chemograph construction. Collection and analysis of water samples from both ASREC and WQFS drains and ASREC stream stations will continue. In addition, a subset of samples from both drains and streams are being analyzed for pH, EC, nitrates, phosphate, and dissolved organic carbon; and a subset of stream samples are being analyzed for pesticides used on the associated fields. We also will begin to compile chemical and flow data in an ACCESS database along with management activities, landscape details, and weather conditions. We will extend our TBA metabolite persistence study to include a range of temperature and moisture conditions of environmental relevance. We will continue to sample fish and turtles from streams and ponds receiving runoff from fields treated with animal manure. In collaboration with USEPA Duluth, hormone concentrations also will be measured using radioimmunoassay in each of the plasma samples.
Journal Articles:
No journal articles submitted with this report: View all 49 publications for this projectSupplemental Keywords:
endocrine disrupting chemicals, ecological effects, aquatic indicators, fish gonadal changes, sex differentiation, ecosystem health risks, manure, effluent irrigation, subsurface injection, broadcasting, androgens, estrogens, trenbolone, trendione, testosterone, estradiol, estrone, estriol, degradation, discharge, chemical transport, soil, hormones, RFA, Scientific Discipline, Health, Environmental Chemistry, Endocrine Disruptors - Environmental Exposure & Risk, endocrine disruptors, Endocrine Disruptors - Human Health, CAFOs, EDCs, endocrine disrupting chemicals, animal feeding operations, concentrated animal feeding operationsProgress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.