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Grantee Research Project Results

2011 Progress Report: An Integrated Approach to Developing a Total Facility Estrogen Budget at a Swine Farrowing CAFO

EPA Grant Number: R833420
Title: An Integrated Approach to Developing a Total Facility Estrogen Budget at a Swine Farrowing CAFO
Investigators: Kullman, Seth W. , Linden, Karl G. , Reckhow, Kenneth H. , Meyer, M. T.
Current Investigators: Kullman, Seth W. , Meyer, M. T. , Reckhow, Kenneth H.
Institution: Nicholas School of the Environment and Earth Sciences , Organic Geochemistry Research Laboratory , Pratt School of Engineering , North Carolina State University
Current Institution: North Carolina State University , Nicholas School of the Environment and Earth Sciences , Organic Geochemistry Research Laboratory
EPA Project Officer: Aja, Hayley
Project Period: February 1, 2008 through January 31, 2013 (Extended to January 31, 2014)
Project Period Covered by this Report: February 1, 2011 through January 31,2012
Project Amount: $663,532
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:

Little information is available regarding the concentration, release, fate and transport of estrogenic compounds in animal waste treatment and storage facilities. In this project we develop quantitative information regarding: reproductive status and estrogen excretion by individual animals, the stability of estrogens in open pit holding lagoons and mobility of estrogens to surface waters following spray field application of swine waste as fertilizer. To address these data gaps, we are focusing on a swine farrowing concentrated animal feeding operation (CAFO) based on its operational units. By creating a hierarchical structure, assessments of estrogen fate will be “parameterized” and used for input into a Bayesian network model.

Progress Summary:

Two primary aims were described in our original proposal.

The objective of Aim 1 is to establish “total facility estrogen budget” based upon composite measurements of natural estrogenic compounds throughout a swine farrowing (CAFO). This aim is comprised of three tasks, which are described below.
 
The objective of Task A is to determine total animal estrogen output from differing operational phases within the swine CAFO. Urine and feces samples were collected from individual gilts and sows at the aforementioned four stages of reproduction (B, G1, G or F). Extracts in ethanol were prepared from the urine and feces using solid phase extraction, and all sample analyses were performed using these extracts. Sample analysis includes the determination of specific estrogen concentrations using LC/MS-MS and estrogenic activity using the yeast estrogen screen (YES) bioassay. To date, LC/MS-MS analysis has been focused primarily on the parent estrogens estrone (E1), 17α-estradiol (E2α), 17β-estradiol (E2β), and estriol (E3). E1 was found to be the most ubiquitous form of estrogen in both the urine and the feces, as it was detected in the vast majority of samples. An overall pattern of increased estrogen output during pregnancy and farrowing was observed across all rounds of sampling. When animals are grouped into respective reproductive phases we observed a defined gradient of estrogenic activity in both the urine and the feces.  Animals that were not confirmed pregnant displayed relatively low estrogenic activity in their waste, whereas estrogenic activity of the waste rose dramatically and reached a peak during gestation and farrowing. Overall, the results from Task A indicate that the urine and feces of the swine in this farrowing operation have appreciable estrogenic activity, as indicated by the YES assay. This activity is attributable to the presence of natural estrogens in the animal waste. Animals in the later stages of pregnancy (gestation, farrowing) had greater estrogen concentrations and estrogenic activity in their waste, indicating that the reproductive stage of the swine is a primary determinant of the estrogen output of a farrowing operation.
 
The objective of Task B is to thoroughly characterize the stability and fate of estrogens in the holding lagoons. Lagoon slurry samples were taken at three different depths (6 inches below the surface, 2 feet below the surface, 6 inches above the top of the sludge), and sludge samples were taken at a level of 1 foot below the top of the sludge.  Samples were collected from eight coordinates on the lagoon, intended to create a representative cross-section of the site.  Samples were collected in winter, spring and summer to assess for seasonal effects. In processing the slurry and sludge samples, each sample was separated into a liquid and a solid fraction by centrifugation, and these fractions were analyzed separately for estrogen content and estrogenic activity.  Trends in lagoon estrogen concentrations were found to be dissimilar over time suggesting possible seasonal effects in estrogen concentrations.  E1 was by far the most abundant form of natural animal estrogen present in the lagoon, with concentrations averaging an order of magnitude greater than concentrations of E2α, E2β, or E3.  Partitioning of estrogens to the solids in the lagoon was evident in both the lagoon slurry and sludge, as was expected based on the moderately lipophilic nature of these compounds (log Kow = 2.6 to 4). Estrogen concentrations in lagoon solids were consistently several hundred-fold greater than concentrations in the liquid fraction of these samples. Consistent with the LC/MS-MS results, YES analysis of slurry samples demonstrated relatively homogenous estrogenic activity throughout the lagoon slurry, higher estrogenic activity in lagoon solids compared to lagoon liquids, and variations in estrogenic activity across the two seasons. 
 
The objective of Task C is to measure estrogen concentration and stability of estrogens in field plots receiving lagoon waste as fertilizer.  Because the lagoon slurry is land-applied as a fertilizer on crop fields, the most likely means of off-site transport of estrogens from a CAFO is via runoff or leaching from these crop fields into groundwater or adjacent surface water. Extensive soil sampling was conducted on waste applied crop fields, which are plowed seasonally and planted with rotating crops of corn, soy, and wheat.  Sampling was conducted over the course of several months in conjunction with spray field applications. LC/MS-MS results indicate that E1 is the only natural animal estrogen that is present at detectable levels in the crop field soil following land application. Additionally, soil extracts were found to be below the limit of detection of the YES assay, so the estrogenic activity of the soil was assessed using a different in vitro bioassay, the T47D-KBluc. The T47D-KBluc is a human breast epithelial carcinoma cell line that has been stably transfected with three estrogen responsive elements upstream from a luciferase reporter. Prior to the land application of lagoon slurry no animal estrogens were detectable in the soil. On the day of land application, levels of E1 in the top 6"” of soil rose to an average of 1300 ng/kg (soil dry weight), and remained at 1300 ng/kg on the day following land application. Over the course of 7 days, E1 concentrations fell to 350 ng/kg and 139 ng/kg in the subsequent 2 months without further land application of lagoon slurry. Data for the T47D-KBluc assay were comparable to LC/MS/MS values.
 
The objective of Aim 2 is to develop an Integrated Bayesian Network Development for Predicting the Fate and Transport of Swine Borne Estrogens.
 
 For Aim 2, we have developed and applied integrated Bayesian networks (BNs) models that can be used to better characterize and assess the generation, fate, and transport of site-specific swine CAFO-derived estrogen compounds. The developed models can be used as decision support tools towards estrogen risk assessment. Modularized and melded BN approaches were used to capture the predictive and casual relationships of the estrogen budget and its movement within and between the three major systems of a swine farrowing CAFO. These systems include the animal barns, the anaerobic waste lagoon, and the spray fields. For the animal barn system, a facility-wide estrogen budget was developed to assess the operation-specific estrogen excretion, using an object-oriented BN (OOBN) approach. The developed OOBN model provides a means to estimate and predict estrogen fluxes from the whole swine facility in the context of both estrogen type and animal operating unit. It also accounts for the uncertainties in the data and in our understanding of the system. Next, mass balance melding BN models were developed to predict the natural estrogen fates and budgets in two lagoon compartments, the slurry and the sludge storage. This involved utilizing mass balance equations to account for the mechanisms of flushing, sorption, transformation, settling, and burial reactions of estrogen compounds in the slurry and sludge storages. As an alternative approach, a regression based BN melding approach was developed to characterize both estrogen fate and budgets as a result of the sequential transformation processes between natural estrogen compounds and to assess the seasonal effects on the estrogen budgets in the two different lagoon compartments. Finally, a dynamic BN model was developed to characterize rainfall-driven estrogen runoff processes from the spray fields. The dynamic BN models were used to assess the potential risk of estrogen runoff to adjacent waterways. In addition, the dynamic model was used to quantify the effects of manure application rates, rainfall frequency, the time of rainfall and irrigation, crop types, on farm best management practices, seasonal variability, and successive rainfall and manure application events on the estrogen runoff. The model results indicate that the farrowing barn is the biggest contributor of total estrogen as compared to the other two operating components. Once the estrogen reaches the anaerobic lagoon, the settling and burial reactions were shown to be the most significant factors controlling estrogen levels in the slurry and sludge, respectively. The estrogen budgets in the lagoon also were found to vary by season, with higher slurry and sludge estrogen levels in the spring as compared to the summer. Nevertheless, the risk of estrogen runoff was predicted to be lower in the spring as compared to the summer, primarily due to the spray field crop management plans adopted. The results also indicated that Bermuda grass performed more favorably when compared to soybean, when it came to retaining surface water runoff in the field. Model predictions indicated that there is a low risk of estrogen runoff losses from the spray fields under multiple irrigation and rainfall events, unless the time interval between irrigation was less than 10 days and/or in the event of a prolonged high magnitude rainstorm event. Overall, the estrone was the most persistent form of natural estrogens in the three major systems of the swine farrowing CAFO.

Future Activities:

Future activities include completion of all sample processing, analytical analysis and bioanalytical analysis. Analysis of all estrogen conjugates, including sulfate and glucuronide estrogen conjugates and plant-derived phytoestrogens, is in progress.  We previously have analyzed a subset of our urine, barn flush and lagoon samples for these compounds and found them to be in abundance. The analysis of all of the samples for this suite of compounds is expected to be completed by spring of 2013.  We also hope to further explore the fate and transport of phytoestrogens on the CAFO by extracting and analyzing the animals'’ feed for the presence of these compounds.


Journal Articles on this Report : 3 Displayed | Download in RIS Format

Publications Views
Other project views: All 28 publications 8 publications in selected types All 8 journal articles
Publications
Type Citation Project Document Sources
Journal Article Lee B, Kullman SW, Yost E, Meyer MT, Worley-Davis L, Williams CM, Reckhow KH. A Bayesian network model for assessing natural estrogen fate and transport in a swine waste lagoon. Integrated Environmental Assessment and Management 2014;10(4):511-521. R833420 (2011)
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  • Journal Article Yost EE, Meyer MT, Dietze JE, Meissner BM, Worley-Davis L, Williams CM, Lee B, Kullman SW. Comprehensive assessment of hormones, phytoestrogens, and estrogenic activity in an anaerobic swine waste lagoon. Environmental Science & Technology 2013;47(23):13781-13790. R833420 (2011)
    R833420 (Final)
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  • Journal Article Yost EE, Meyer MT, Dietze JE, Williams CM, Worley-Davis L, Lee B, Kullman SW. Transport of steroid hormones, phytoestrogens, and estrogenic activity across a swine lagoon/sprayfield system. Environmental Science & Technology 2014;48(19):11600-11609. R833420 (2011)
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  • Supplemental Keywords:

    CAFO, estrogens, fate and transport, mass balance

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    28 publications for this project
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