EVALUATING THE PHYSICAL AND BIOLOGICAL AVAILABILITY OF PESTICIDES AND CONTAMINANTS IN AGRICULTURAL ECOSYSTEMS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 0229487
• Multistate No.: W-2082
• Project Start Date: Jul 1, 2012
• Project End Date: Sep 30, 2015
• Program Code: [(N/A)]- (N/A)

Recipient Organization
VIRGINIA POLYTECHNIC INSTITUTE
(N/A)
BLACKSBURG,VA 24061

Performing Department
Crop & Soil Environmental Sciences

Non Technical Summary
Chemicals associated with manure from confined animal feeding operations (CAFOs) are of concern for both surface and groundwater quality, driving the need for data on factors influencing their input and subsequent fate in the environment. Although many investigations have reported frequent detections of sex steroid hormones in excreta of farm animals, there is a lack of systematic tracking of the occurrence of those compounds in waste generated at different life stages of animals, especially those raised in CAFOs, making it difficult to accurately assess the total mass flux of sex hormones from CAFOs to the environment. A better characterization of the occurrence of sex hormones in the wastes from CAFO sites at differing life stages will help the producers and regulators assess potential "hot spots" within production facilities for hormone input to the environment and, therefore, will help concentrate management, regulation,and remediation efforts on the most problematic areas of CAFOs. In addition, there is also a lack of data on the fate of those compounds during collection, storage, and treatment of animal waste, as well as in soil and water systems that are impacted by animal waste. While more data is needed to understand the fate of these hormones at their source of production, further data is needed to fully characterize their behavior once they reach other terrestrial or aquatic environments. The effect of various environmental factors such as soil type, organic matter content, type of minerals, soil moisture and temperature, sunlight, etc. on the fate of sex hormones are also poorly understood. This project will generate extensive data on the effects of poultry and swine CAFO production and waste management practices on hormone/metabolite levels in animal waste. Ultimately this data will be critical to identifying production sites that pose the greatest potential to generate waste with high hormone levels. It will illuminate which waste management practices will be most effective at reducing the potential of hormones to contaminate off-site water resources. This project will provide extensive and detailed environmental fate data on individual hormones that will allow risk assessors to predict their runoff and/or leaching potential to surface and groundwater, and identify exposure routes for human and wildlife populations to animal hormones.

Animal Health Component

(N/A)

Research Effort Categories

Basic

(N/A)

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
102	0110	2000	100%

Knowledge Area
102 - Soil, Plant, Water, Nutrient Relationships;

Subject Of Investigation
0110 - Soil;

Field Of Science
2000 - Chemistry;

Keywords

animal hormones

confined animal feeding operations

environmental fate

occurrence

persistence

transformation

animal waste

Goals / Objectives
To identify and quantify fundamental chemical, physical, and biological processes relevant to pesticides and contaminants in agricultural ecosystems, To evaluate existing transport models for predicting the fate and transport of pesticides and contaminants in agricultural ecosystems, To provide information and outreach required for field-scale recommendations for the management of pesticides and contaminants in agricultural, suburban, x-urban and rural ecosystems.

Project Methods
A comprehensive investigation on the occurrence of sex hormones and their conjugates in animal waste generated at different life stages of animals at different CAFO production sites are conducted. Animal waste from different production sites of a swine CAFO and a poultry CAFO were collected and analyzed for estrone,17 alpha-estradiol, 17 beta-estradiol, testosterone, progesterone, and their glucuronide or sulfate conjugates. Both laboratory and on-site studies were conducted. Animal waste were collected during the second part of the waste management system which consists of collection, storage,treatment, and transfer and land application. Target sex hormone compounds were analyzed and their occurrence in animal waste handled with different options at each management function were compared. The laboratory experiments for Objective 2 will be conducted to understand Effect of different animal waste storage on the fate of hormones and their conjugates will be conducted. The results from the laboratory studies will be compared with the results from the on-site investigation. Aquatic and terrestrial environmental fate of target compounds will be modeled after US EPA Subdivision N guidelines for the registration of pesticides. We will measure the rate of degradation of each active compound in aquatic hydrolysis, photolysis, and water/sediment degradation tests followed by identification and assessment of the stability of primary degradation products formed in each test. For the terrestrial fate experiments, major soil and environmental factors that may affect the fate of compounds of interest will be evaluated. The major soil and environmental factors to be evaluated are soil organic matter, soil minerals, soil moisture, soil pH, ambient temperature, and sunlight. The soil and sediment adsorption coefficients, aerobic degradation rates, and surface soil photolysis rates of target compounds will be measured. The results generated from those experiments will help the regulators and producers better assess and predict the fate of sex hormones and their conjugates at a specific land application site and the potential input of those compounds to the associated aquatic systems. The data generated from this project will be presented at professional meetings and workshops. The data will be shared with CAFO producers. Results will be published in high quality peer-reviewed scientific journals. We will provide USDA with data generated from this project to assist policy making at the federal level.

Progress 07/01/12 to 09/30/15

Outputs
Target Audience:Environmental scientists, food producers, pesticides producers, general public, decision makers, animal production facility managers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One postdoc andthree graduate studentslearned analytical techniques for pesticides analysis using state-of-the-art chromatographyinstrument. The students gained experience in presenting scientific findings at international meetings. Three undergraduate students gained research experience by working with the graduate students. How have the results been disseminated to communities of interest?The finding of this work was presented at 2012,2013, 2014, and 2015ASA-CSSA-SSSA International Annual Meetings. Published six peer-reviewed articles in fiveinternational scientific journals. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Thiamethoxam (TMX) and Clothiniadin (CLO) are second-generation neonicotinoid insecticides commonly used as seed coatings for a variety of crops. Neonicotinoids are highly soluble and, therefore, easily taken up by young plants from coated seeds to provide protection for seeds and young plants from insects damage. Recent investigations have detected neonicotinoids in aquatic environment surrounding agricultural fields planted with neonicotinoid coated corn and canola seeds. It was found that the neonicotinoids stream concentrations pulsed with rainfall events during crop planting of the surrounding fields, suggesting treated seeds as a likely source of neonicotinoids in the aquatic environment. Neonicotinoids have been found to be toxic to certain non-target aquatic even at concentrations as low as 0.01 ppb. Therefore, the objective of this research was to determine if it is possible for TMX and CLO to move from corn seeds to the surrounding soilenvironment. TMX and CLO-coated corn seeds were planted in pots in a greenhouse and the plants were harvested at V4 growth stage. The levels of TMX and CLO in bulk soil, root soil, and rhizosphere soil, and in plant roots and shoots were analyzed using liquid chromatography/tandem mass spectrometry (LC/MS/MS). Results from this study demonstrated that both TMX and CLO coated on corn seeds moved, within 3 weeks of planting, into the surrounding soil in addition to being taken up by the plants. This data suggested that seed coating pesticides have the potential to migrate to the surrounding soil and aquatic environment during the growing season. Antibiotics and hormones associated with animal manure are of increasing environmental concern due to the potential for increased antibiotic resistance and endocrine disruption in aquatic organisms after land application of manure. Manure application technology may affect the environmental behavior of these manure-borne emerging contaminants. Therefore, rainfall simulations were conducted on plots receiving three manure treatments (surface application, subsurface injection, and no manure control) to determine the fate and transport of animal hormones, hormone conjugates, and four antibiotics commonly used in poultry and dairy production. Compared to antibiotics, animal hormones and their conjugates tend to accumulate in the top 0-5 cm of soil, while some antibiotics migrated down to 5-20 cm soil. Comparing to manure surface application, subsurface injection reduces the overall distribution of hormones and antibiotics on land surface, resulting in significantly less surface runoff losses of those compounds by as high as97%. Thus, injection could be considered a best management practice to prevent loss of antibiotics in surface runoff. The development of models for understanding antibiotic resistance gene (ARG) persistence and transport is a critical next step toward informing mitigation strategies to prevent the spread of antibiotic resistance in the environment. A field study was performed using a mass balance approach to gain insight into the transport and dissipation of ARGs following land application of manure. Soil from a small drainage plot including a manure application site, an un-manured control site, and an adjacent stream and buffer zone were sampled for ARGs and metals before and after application of dairy manure slurry and a dry stack mixture of equine, bovine, and ovine manure. Results of mass balance suggest growth of bacterial hosts containing ARGs and/or horizontal gene transfer immediately following slurry application with respect to ermF, sul1, and sul2, and following a lag (13 days) for dry stack amended soils. Generally no effects on tet(G), tet(O), or tet(W) soil concentrations wereobserved despite presence of these genes in applied manure. Dissipation rates were fastest for ermF in slurry treated soils (logarithmic decay coefficient of -3.5) and for sul1 and sul2 in dry stack amended soils (logarithmic decay coefficients of -0.54 and -0.48, respectively) and evidence for surface and subsurface transport was not observed. Results provide a mass balance approach for tracking ARG fate and insights to inform modeling and limiting the transport of manure-borne ARGs to neighboring surface water. Pirlimycin, a lincosamide antibiotic, and cephapirin, a cephalosporin antibiotic are among the most commonly used antibiotics used by the majority of dairy farms in the US. Assessment of pirlimycin and cephapirin loading to the environment via fecal and urinary excretion is critical to develop efficient management strategies to reduce environmental pollution by the livestock industry. Therefore, analytical methods to identify and quantify pirlimycin and cephapirin in bovine feces and urine were developed. A novel approach was developed to quantify estrone (E1), 17beta-estradiol (bE2), 17a-estradiol (aE2), estriol (E3), and 17alfa-ethynylestradiol (EE2) in storm water runoff and swine lagoon wastewater samples. A considerable amount of residue was collected when lagoon wastewater samples were centrifuged therefore both resulting residues and aqueous portions were analyzed separately. A modified Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method was utilized to efficiently extract the target analytes in the residue. Aqueous portions were pre-concentrated using solid phase extraction (SPE). A simple liquid-liquid extraction in a test tube was carried out as a sample clean-up step after SPE. This approach is inexpensive and requires only 3 mL of organic solvent per analysis.

Publications

Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Scientific community, policymakers, general public. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? One postdoctoral researcher, three graduate students, and 2 undergraduate students have been trained. How have the results been disseminated to communities of interest? Presentations at the 2013 ASA-CSSA-SSSA International Annual Meetings. Published five peer-reviewed articles in four international scientific journals. What do you plan to do during the next reporting period to accomplish the goals? Generate three international meeting presentations and three peer-reviewed journal articles.

Impacts
What was accomplished under these goals? The development of models for understanding antibiotic resistance gene (ARG) persistence and transport is a critical next step toward informing mitigation strategies to prevent the spread of antibiotic resistance in the environment. A field study was performed using a mass balance approach to gain insight into the transport and dissipation of ARGs following land application of manure. Soil from a small drainage plot including a manure application site, an un-manured control site, and an adjacent stream and buffer zone were sampled for ARGs and metals before and after application of dairy manure slurry and a dry stack mixture of equine, bovine, and ovine manure. Results of mass balance suggest growth of bacterial hosts containing ARGs and/or horizontal gene transfer immediately following slurry application with respect to ermF, sul1, and sul2, and following a lag (13 days) for dry stack amended soils. Generally no effects on tet(G), tet(O), or tet(W) soil concentrations were observed despite presence of these genes in applied manure. Dissipation rates were fastest for ermF in slurry treated soils (logarithmic decay coefficient of -3.5) and for sul1 and sul2 in dry stack amended soils (logarithmic decay coefficients of -0.54 and -0.48, respectively) and evidence for surface and subsurface transport was not observed. Results provide a mass balance approach for tracking ARG fate and insights to inform modeling and limiting the transport of manure-borne ARGs to neighboring surface water. Pirlimycin, a lincosamide antibiotic, and cephapirin, a cephalosporin antibiotic are among the most commonly used antibiotics used by the majority of dairy farms in the US. Assessment of pirlimycin and cephapirin loading to the environment via fecal and urinary excretion is critical to develop efficient management strategies to reduce environmental pollution by the livestock industry. Therefore, analytical methods to identify and quantify pirlimycin and cephapirin in bovine feces and urine were developed. For Pirlimycin and cephapirin, samples were extracted with methanol-phosphate buffer and cleaned up by SPE before their analysis using UPLC-MS/MS. The method for pirlimycin had LOQ of 1.47 ng/g wet feces and 0.90 ng/mL urine, recovery of 80-108%, repeatability of 2.3-13%, and reproducibility of 2.3-14% for both bovine feces and urine. The method for cephapirin had LOQ of 4.02 ug/kg and 0.96 ug/L for feces and urine, respectively, recovery of > 60% for fecal samples and >80% for urine samples, and intra- and inter-day variation <10%. Both methods were robust, sensitive, and accurate and can be used to assess the fate and environmental impact of antibiotics used on farms. Pirlimycin was detected at 40.5-287 ng/g and 46.1-254 ng/mL in feces and urine, respectively, 10 - 120 h following intramammary dosing. Cephapirin was primarily excreted via urine and its urinary excretion was influenced by day (P = 0.03). Peak excretion (2.69 mg) was on day 1 following intramammary infusion and decreased sharply thereafter (0.19, 0.19, 0.08, and 0.17 mg on day 2, 3, 4, and 5, respectively) reflecting a quadratic pattern of excretion (Quadratic: P = 0.03). This data will help develop manure segregation and treatment methods to minimize the risk of antibiotic loading to the environment from dairy farms. A novel approach was developed to quantify estrone (E1), 17beta-estradiol (bE2), 17a-estradiol (aE2), estriol (E3), and 17alfa-ethynylestradiol (EE2) in storm water runoff and swine lagoon wastewater samples. A considerable amount of residue was collected when lagoon wastewater samples were centrifuged therefore both resulting residues and aqueous portions were analyzed separately. A modified Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method was utilized to efficiently extract the target analytes in the residue. Aqueous portions were pre-concentrated using solid phase extraction (SPE). A simple liquid-liquid extraction in a test tube was carried out as a sample clean-up step after SPE. This approach is inexpensive and requires only 3 mL of organic solvent per analysis. The resulting extracts were further purified using a dispersive solid phase extraction (dSPE) technique (not required in storm water analyses). The bE2 and E1 glucuronide and sulfate conjugates were also analyzed in the aqueous portions of the lagoon samples as well as in the storm water runoff samples. An enzymatic hydrolysis was performed prior to SPE to deconjugate conjugated estrogens. Samples were derivatized with dansyl chloride to enhance LC/MS/MS analytical sensitivity. Insignificant matrix effects (1-12%) were determined for the analyses of aqueous samples. Thus, matrix matched calibration curves were not required. Matrix effects for residue analyses ranged from 14 to 20%, requiring that matrix matched calibration curves be used for quantification. All developed methods gave 84-106% recoveries for free estrogens and 65-86% recoveries for estrogen conjugates. The LODs for lagoon wastewater and storm water analyses ranged from 0.9-2 ng/L and 0.3-0.5 ng/L, respectively. The developed methods were validated by analyzing eighteen lagoon water samples and twenty six storm water samples in three replicates.

Publications

• Type: Journal Articles Status: Published Year Published: 2013 Citation: Fahrenfeld, N., K. Knowlton, L. A. Krometis, W. C. Hession, K. Xia, E. Lipscomb, K. Libuit, B. L. Green, A. Pruden. 2013. Effect of Manure Application on Abundance of Antibiotic Resistance Genes and their Attenuation Rates in Soil: Field-Scale Mass Balance Approach. Environ. Sci. Technol. 48:26432650
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Gunatilake, S. R., J. W Kwon, T. E. Mlsna, and K. Xia*. 2014. A novel approach to determine estrogenic hormones in swine lagoon wastewater using QuEChERS method combined with solid phase extraction, and LC/MS/MS analysis. Anal. Methods. 6:9267 - 9275.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Ray, P., K.F. Knowlton, C. Shang, and K. Xia. 2014. Development and validation of a UPLC-MS/MS method to monitor cephapirin excretion in dairy cows following intramammary infusion. PLOS ONE. 9:1-12.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Ray, P., K.F. Knowlton, C. Shang, and K. Xia. 2014. Method development and validation: solid phase extraction (SPE)-ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) quantification of pirlimycin in bovine feces and urine. J AOAC International. 97:1730-1736.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: K. Xia, C Qin, D. Troya, C. Shang. Surface catalyzed polymerization of emerging contaminants by Fe(III)-modified montmorillonite. 2014 International Symposium on Environment and Health (ISEH 2014). Beijing, China. July 4-5 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: K Xia, L Hundal. Occurrence and fate of emerging contaminants in biosolids and biosolids-amended soils. 2014 International Symposium on Environment and Health (ISEH 2014). Beijing, China. July 4-5 2014.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Sosienski, T., K. Xia, R. O. Maguire, and S. Kulesza. Fate of Hormones in Surface-Applied and Subsurface Injected Poultry Litter. ASA-CSSA-SSSA International Annual Meetings, Tampa, FL. Nov. 3-6, 2013.

Progress 10/01/12 to 09/30/13

Outputs
Target Audience: scientists decision/policy makers growers/producers general public Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Three graduate students and four undergraduate students have been trained. How have the results been disseminated to communities of interest? Presentations at the 2012 and 2013ASA-CSSA-SSSA International Annual Meetings. Publisd five peer-reviewed articles in four international scientific journals What do you plan to do during the next reporting period to accomplish the goals? Generate three international meeting presentations and three peer-reviewed journal articles.

Impacts
What was accomplished under these goals? Estrone, 17β-estradiol, and their conjugates were frequently detected in chicken litters and swine manures at levels ranging from low ng/g to several hundreds ng/g. The levels of conjugated hormones in those samples were at similar or higher levels than free hormones, suggesting the importance of studying the fate and impact of hormone conjugates in the environment. Testosterone levels in both swine and poultry waste were more than 100 times lower than its precursors, suggesting the importance of understanding the environmental fate of those precursors. Laboratory incubation study demonstrated that 17β-estradiol-3-glucuronide(E2-3G) transformation followed 1st order kinetics and was largely biologically driven at low temperature. With increasing environmental temperature, abiotic transformation of E2-3G also becomes significant, resulting in faster transformation rate. Our recent study has demonstrated that Fe(III)-modified montmorillonite quickly (within minutes) polymerize 17-beta estradiol into dimers, trimers, tetramers, or even pentamers, which are hundreds to thousands time less water soluble than 17-beta estradiol. The less water soluble a compound is, the less bioavailable and toxic it is to organisms. This study suggests the potential of using Fe(III)-modified montmorillonite as a low cost material for removing hormones from animal waste.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sosienski, T., K. Xia, R. O. Maguire, and S. Kulesza. Fate of Hormones in Surface-Applied and Subsurface Injected Poultry Litter. ASA-CSSA-SSSA International Annual Meetings, Tampa, FL. Nov. 3-6, 2013.
• Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Guo, H., and K. Xia. 2012. Fate of Hormone Conjugates in Soils. ASA-CSSA-SSSA International Annual Meetings, Cincinnati, OH. Oct. 22, 2012.
• Type: Journal Articles Status: Under Review Year Published: 2013 Citation: Fahrenfeld, N., K. Knowlton, L. A. Krometis, W. C. Hession, K. Xia, E. Lipscomb, K. Libuit, B. L. Green, A. Pruden. 2013. Effect of Manure Application on Abundance of Antibiotic Resistance Genes and their Attenuation Rates in Soil: Field-Scale Mass Balance Approach. Environ. Sci. Technol. (in review).
• Type: Journal Articles Status: Published Year Published: 2013 Citation: Gunatilake, S. R., S. Steelhammer, J. W. Kwon, J. Rodriguez, K. Xia, K. Armbrust, and T. E. Mlsna. 2013. Analysis of Estrogens in Wastewater using solid phase extraction, the QuECHERS cleanup, and liquid chromatography tandem mass spectrometry. J. AOAC International. 96: 1440-1447.
• Type: Journal Articles Status: Published Year Published: 2012 Citation: Keith A. Maruya, D. E. Vidal-Dorsch, S. M. Bay, J. W. Kwon, K. Xia, and K. L. Armbrust. 2012. Organic contaminants of emerging concern in sediments and flatfish collected near outfalls discharging treated wastewater effluent to the Southern California Bight. Environ. Toxicol. Chem. 31:26832688.
• Type: Journal Articles Status: Published Year Published: 2012 Citation: Xia, K.*, G. Hagood, C. Childers, J. Atkins, B. Rogers, L. Ware, K. Armbrust, J. Jewell, D. Diaz, N. Gatian, and H. Folmer. 2012. Polycyclic Aromatic Hydrocarbons (PAHs) in Mississippi Seafood from Areas Affected by the Deepwater Horizon Oil Spill. Environ. Sci. Technol. 46:53105318.
• Type: Journal Articles Status: Published Year Published: 2012 Citation: Diehl, J., S. E. Johnson, K. Xia, A. West, and L. Tomanek. 2012. The distribution of 4-nonylphenol in marine organisms of North American Pacific Coast estuaries. Chemosphere. 87:490497.
• Type: Journal Articles Status: Published Year Published: 2012 Citation: Kwon, J. W. and K. Xia*. 2012. Fate of Triclosan and Triclocarban in Soil Columns With and Without Biosolids Surface Application. Environ. Toxicol. Chem. 31:262269.

Progress 10/01/11 to 09/30/12

Outputs
OUTPUTS: Animal hormones and antibiotics associated with manure from confined animal feeding operations and pharmaceutical and personal care compounds frequently detected in biosolids are of concern for their negative impact on environmental quality due to land application of both animal manure and biosolids. This concern drives the need for data on factors influencing their input and subsequent fate in the environment. For this project the following research activities were performed and are currently underway: 1) investigating the occurrence of hormones and their glucuronide and sulfate conjugates in animal waste generated at different life stages of poultry and swine production; 2) investigating how different animal waste management practices affect the fate of hormones and their conjugates in animal wastes through both laboratory and on-site investigations; 3) determining the aquatic and terrestrial fate of hormones, hormone conjugates, antibiotics, and pharmaceutical and personal care compounds in laboratory studies similar to those used for generating environmental fate data for pesticides. Samples generated at different life stages of animals were collected at a swine production facility and analyzed for estrone, 17alpha-estradiol, 17beta-estradiol, testosterone, progesterone, and their glucuronide or sulfate conjugates using liquid chromatography-mass spectrometry (LC/MS/MS). The same compounds were also analyzed for chicken litter samples collected from different poultry facilities throughout the State of Mississippi. One graduate student was trained on analytical method development for hormones and their conjugates using state of the art LC/MS/MS. Research results were communicated with the swine production facility managers. The results were presented at four national meetings, included in one submitted manuscript and a Ph.D thesis, and will be included in one manuscript in preparation. Samples were collected from sites with different animal waste management practices and analysis is currently conducted by a master degree student. Laboratory experiments mimicking different animal waste management practices are currently conducted by the same student and a postdoctoral researcher to investigate transformation kinetics of target compounds. Soil samples from different depth and surface runoff and leachate of fields receiving poultry litter application are collected before and after litter application. The results were presented at one national meeting and will be included in a manuscript in preparation. A laboratory soil column study was conducted to investigate the transport and transformation of antibacterial compounds, triclosan (TCS) and triclocarban (TCC) in a biosolids-surface applied soil. During this study, the column leachates and soil samples were analyzed for TCS, TCC, and their transformation products. The result of this study was published in Environmental Toxicology and Chemistry, a leading peer-reviewed environmental journal. PARTICIPANTS: Jeong-Wook Kwon (Research Scientist), Mississippi State University; Kevin Armbrust (Associate Professor, co-investigator), Mississippi State University; Mark Crenshaw (Professor, co-investigator), Mississippi State University; Kusum Verma (Ph.D student), Mississippi State University; Theresa Sosienski (MS Student), Virginia Tech; Huiqin Guo (postdoctoral researcher), Virginia Tech; Rory Maguire (Associate Professor, collaborator), Virginia Tech TARGET AUDIENCES: Policy makers, regulators, producers, general public, scientific community, PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
A new sample extraction technique was developed for analysis of hormones and their conjugates in animal waste. This technique significantly shortened the analytical time per sample and decreased the amount of extraction solvent needed per sample. Estrone, 17 beta-estradiol, and their conjugates were frequently detected in the chicken litter at levels ranging from low ng/g to several hundreds ng/g. Those compounds were at similar levels in the swine manure sample. The levels of conjugated hormones in those samples were at similar or higher levels than free hormones, suggesting the importance of studying the fate and impact of hormone conjugates in the environment. Testosterone levels in both swine and poultry waste were more than 100 times lower than its precursors (1,4-androstadiene-3,17-dione and 4-androstene-3,17-dione), suggesting the importance of understanding the environmental fate of those precursors. Laboratory incubation study for two soils with different physic-chemical characteristics demonstrated that 17 beta-estradiol-3-glucuronide(E2-3G) transformation follows 1st order kinetics and was largely biologically driven at low temperature. With increasing environmental temperature, in addition to biotic transformation abiotic transformation of E2-3G also becomes significant. 17 beta-estradiol and estrone were major metabolites of E2-3G biotic and abiotic transformation, indicating deconjugation coupled with oxidation of 17th hydroxyl functional group were major mechanisms for E2-3G transformation. In unsterilized soils, after their concentrations reached the highest, both 17 beta-estradiol and estrone transformed rapidly within 24 hours followed by slower transformation. At low temperature, ~15% of both compounds remained in the soils at the end of 24-day incubation. This study suggests that conjugated hormones can be a potential source of free hormones in soils with high clay and organic matter contents at low temperature. The laboratory column study has shown surface application of biosolids significantly retarded TCS and TCC transformation. Downward movement of TCS and TCC occurred within 10-cm soil depth. Methyl-TCS was not detectable in the leachates but was detected in the top 5-cm soil layer, with more appeared in the biosolids-applied soil. At the end of the column study, carbanilide (CBA) was the only detectable TCC reductive dechlorination products in the soil. None of the TCC reductive dechlorination products were detectable in the leachates. Detection of 3,4-dichloroaniline (3,4-DCA) and 4-chloroaniline (4-CA) suggested the occurrence of TCC hydrolysis. Rapid leaching of 4-CA through the soil column was observed. The 3,4-DCA was detected throughout the entire 20-cm depth of the soil column but not in the leachates. The fact that only small percentages of the transformed TCS and TCC appeared, after the 101-day column study, in the forms of the products analyzed suggested that either the investigated transformation pathways were not significant or rapid transformation of the those products had occurred.

Publications

• Huiqin Guo, and K. Xia. 2012. Fate of Hormone Conjugates in Soils. ASA-CSSA-SSSA International Annual Meetings, Cincinnati, OH. Oct. 22, 2012.
• Keith A. Maruya, D. E. Vidal-Dorsch, S. M. Bay, J. W. Kwon, K. Xia, and K. L. Armbrust. 2012. Organic contaminants of emerging concern in sediments and flatfish collected near outfalls discharging treated wastewater effluent to the Southern California Bight. Environ. Toxicol. Chem. 31:2683-2688.
• Xia, K., G. Hagood, C. Childers, J. Atkins, B. Rogers, L. Ware, K. Armbrust, J. Jewell, D. Diaz, N. Gatian, and H. Folmer. 2012. Polycyclic Aromatic Hydrocarbons (PAHs) in Mississippi Seafood from Areas Affected by the Deepwater Horizon Oil Spill. Environ. Sci. Technol. 46:5310-5318.
• Diehl, J., S. E. Johnson, K. Xia, A. West, and L. Tomanek. 2012. The distribution of 4-nonylphenol in marine organisms of North American Pacific Coast estuaries. Chemosphere. 87:490-497.
• Kwon, J. W. and K. Xia. 2012. Fate of Triclosan and Triclocarban in Soil Columns With and Without Biosolids Surface Application. Environ. Toxicol. Chem. 31:262-269.
• Kelly, J. G., F. X. Han, Y. Su, Y. J. Xia, V. Philips, Z. Q. Shi, D. L. Monts, S. T. Pichardo, and K. Xia. 2012. Rapid Determination of Mercury in Contaminated Soil and Plant Samples Using Portable Mercury Direct Analyzer without Sample Preparation, a Comparative Study. Water Air Soil Pollut. 223:2361-2371.