Source: UNIVERSITY OF NEBRASKA submitted to NRP
DEVELOPMENT OF ADVANCED ANALYTICAL METHODS FOR WATER QUALITY RESEARCH
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0221387
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Feb 1, 2010
Project End Date
Jan 31, 2015
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIVERSITY OF NEBRASKA
(N/A)
LINCOLN,NE 68583
Performing Department
School of Natural Resources
Non Technical Summary
Contamination of water can severely reduce its biological and economic value. Though considerable progress has been made over the last few decades in limiting levels of many types of contaminants, it is clear that chemical contamination of water cannot be prevented. Increasing human populations increase pressure on municipal and residential waste treatment facilities which were not designed to remove synthetic organic contaminants. Treated municipal wastewater therefore releases traces of steroids, prescription and nonprescription pharmaceuticals, illicit drugs, and other chemicals into receiving waters (Kolpin et al 2004; Stackelberg et al 2004). Residues of antibiotics, other veterinary pharmaceuticals, and steroid hormones are known to occur in livestock waste (Burkholder et al 2007; Campagnolo et al 2002; Dolliver and Gupta, 2008; Zheng et al 2008; Shappell et al 2007). Municipal wastewater discharge, run-off from feedlots and crops fertilized with manure are all potential routes for releasing these compounds into aquatic environments. Because hormones are integrally linked to reproductive functions, their occurrence in rivers and lakes are likely to impact the reproductive health of aquatic organisms. The impact of antibiotic residues in agricultural and municipal biosolids to soil and aquatic microorganisms is under close scrutiny in light of the proliferation of antibiotic-resistant bacteria. What is lacking, however, is a clear understanding of the relative importance of these sources and what action can be taken to minimize contamination from these compounds. This research will provide methods for accurate detection and quantification of these contaminants.
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
1120210200050%
1330210200050%
Knowledge Area
133 - Pollution Prevention and Mitigation; 112 - Watershed Protection and Management;

Subject Of Investigation
0210 - Water resources;

Field Of Science
2000 - Chemistry;
Goals / Objectives
Goal. Using advanced analytical methods, identify and characterize the most important sources and types of emerging organic chemicals that may enter water supplies in areas with a high-density of livestock and row-crop agriculture. Objective A. Develop, validate and publish methods for quantitative analysis of veterinary pharmaceuticals, including antibiotics and growth promoters, and natural steroid hormones in source matrices (livestock waste, soils receiving manure, and impacted water). Objective B. Utilize methods developed in Objective A to support field and laboratory research projects focused on identifying and characterizing the concentrations of natural and synthetic steroid hormones and several classes of antimicrobials (e.g.- sulfonamides, tetracyclines, and ionophores) in livestock and municipal waste. Objective C. Identify biologically and abiotically-produced transformation products of steroid hormones and pharmaceuticals using high-resolution (Q-TOF) LC/MS instrumentation to better characterize the environmental fate of parent compounds and their potential for impacting water quality. Outputs and Milestones. Research associated with all of the objectives will be conducted simultaneously. Research results will be reported as required, agencies and other entities supporting specific aspects of the project, in addition to abstracts, proceedings and poster/oral presentations at scientific conferences and workshops. Results will also be communicated to natural resources, environmental, and agricultural agencies and individuals within Nebraska and elsewhere to promote policies and management practices that protect human and environmental health. Manuscripts will be submitted to scientific/professional journals including, but not limited to: the Journal of Environmental Quality, Environmental Science & Technology, Environmental Toxicology & Chemistry, Chemosphere, and Environmental Pollution. Student dissertations (and theses) will be completed during the course of the project. Annual reports of project accomplishments will be submitted to the Director of SNR and to the ARD as required and requested.
Project Methods
Analytical method development generally requires identification of analyte(s) to be measured, review of methods reported in the literature, types of samples to be tested, expected range of concentrations, extraction and purification techniques, instrumental requirements, and required sensitivity. Because LC/MS instrument and operator time can be expensive, multi-residue methods developed around compound classes generally provide the most efficient use of these resources. Although addition of analytes to a method increases the amount of information obtained from a single analysis, variability in extraction recovery may reduce sensitivity for individual compounds in the method. The process of developing and validating a new method are similar and typically follow the steps of literature review, instrument tuning and calibration, adaption of extraction and purification methods, followed by evaluation of the reproducibility and sensitivity for each compound. As an example, a method developed in the Water Sciences Laboratory for measuring antimicrobial compounds in water was based on a method published by Batt et al (2006) for 4 sulfonamides - sulfamethazine, sulfadimethoxine, sulfachloropyridazine, and sulfamethoxine - was expanded to include sulfamethoxazole, sulfamerazine, and sulfathiozole because of the similarity of their chemical properties. The method was modified to allow for automated solid phase extraction (SPE) using a new Spark Holland Symbiosys Environ system interfaced with the Waters Quattro Micro for detection by liquid chromatography-tandem mass spectrometry. The instrument was tuned by infusing each compound separately, choosing and optimizing signal for each compound mass fragmentation transition separately. During method development, an application for analysis of sulfonamide antibiotics using automated on-line SPE was provided by the system manufacturer to assist with method development (Choi et al 2006) and used to help select extraction and elution conditions. The method detection limits were estimated from variability in replicate analysis of 7 low-level fortified (0.005 ng/g) blanks (purified water), and ranged from 0.0004 to 0.0014 ng/g. Recovery of the 7 sulfonamide antimicrobials ranged from 95 to 165% at 0.005 ng/g. The method was applied to several samples of groundwater suspected to be impacted by agricultural wastewater sources. Only 1 compound - sulfamethazine - was detected in the samples.

Progress 02/01/10 to 01/31/15

Outputs
Target Audience:Target audiences for these methods include environmental scientists and engineers, livestock and crop producers, and regulatory agencies concerned with the impact of pharmaceuticals, steroid hormones and growth promoters to water quality. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Twelve (12) graduate students were trained through these projects, in including 3 who went on to coauthor methods papers using advanced analytical tools. Most of the presentations using these methods were given by graduate students at local, regional or national professional society meetings. Two research technologists were trained in these methods, and one has remained on staff to apply these methods to new projects. How have the results been disseminated to communities of interest?The results of these projects have been disseminated through 20+ journal articles published between 2010 and 2015, and through 56 presentations at local, regional, national and international conferences and workshops. Articles were published in Analytical and Bioanalytical Chemistry, Aquatic Toxicology, Journal of Environmental Quality, Journal of Environmental Engineering, Environmental Pollution, Environmental Science and Technology. Environmental Toxocology and Chemistry, Science of the Total Environment, and Transaction of the ASABE. National conferences where these method and result have been presented include American Chemical Society, American Society of Microbiology, American Geophysical Union, American Water Resources Association, Association of Environmental Engineering and Science Professors (AEESP), ASA, CSSA, and SSSA, Emcon 2009, EmCon 2014, Society of Environmental Toxicology and Chemistry, Waste to Worth Conference, 5th International Passive Sampling Workshop (IPSW 2012). Noteworthy products include three separate refereed journal articles in ACS Environmental Science and Technology and three in the ASA Journal of Environmental Quality. Invited presentations were given at special symposiums on antibiotics and antibiotic resistance a the American Chemical Society National meeting (2015) and the ASA, CSSA, and SSSA National Meeting (2015). A workshop on antibiotics and antibiotic resistance in agroecosystems lead to publication of a special issue on this topic to be published in early 2016. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Major activities completed: Twenty three (23) referred journal articles were published during the project period using the advanced analytical methods described described in this research, supporting activites in 10 separate grant-funded research projects. Major research projects ranged from evaluation of the environmental fate and biological effects of steroid hormones in aquatic enviroments (NSF Award 0966850), to improved understanding of persistence and uptake of antibiotic resistant pathogens (USDA NIFA 2011-67019-20052) to determining the effects of management practices on fate and transport of hormones in cattle manures (USEPA STAR Grant #R833423). Specific objectives met: Objective A was met by developing and publishing articles and providing presentations on advanced analytical methods to measure and characterize the most common and persistent steroid hormones and veterinary pharmaceutical residues in livestock waste, soils receiving manure, and impacted aquatic environments. Objective B was met by using or modifying published methods to support field and laboratory research projects focused on identifying and characterizing the concentrations of natural and synthetic steroid hormones and several classes of antimicrobials (e.g.- sulfonamides, tetracyclines, and ionophores) in livestock and municipal waste. Objective C was partially met through identication of biologically and abiotically-produced transformation products of steroid hormones and pharmaceuticals to better characterize the environmental fate of parent compounds and their potential for impacting water quality. Significant results achieved: Development of advanced analytical methods was documented in methods papers (Snow et al 2013 and Sallach et al 2015) and in the research studies investing the occurrence of these compounds (Biswas et al 2012; Yuan et al 2013; van Donk et al 2013; Zhang et al 2013; Joy et al 2013; and others) Application of these methods to the above research settings has provided a means to measure and prorititize the environmental and ecological significance of natural and synthetic steroid hormones and veterinary pharmaceuticals. For example, the most persistent and biologically active steroid hormones in livestock waste are likely to be endogenous estrogens and estrogen metabolites. Uncontrolled runoff from concentrated animal production facilities is of environmental concern regardless of the use of growth promotants (Bartelt et al 2012). Progesterone in manure and wastewater may be transformed into more biologically active androgens. Composting manure is an effective method for reducing steroid hormone concentrations (Bartelt et al 2013). Commonly detected antibiotic residues in livestock waste include tetracylines, macrolides and sulfoamides each with differing mobility and persistence after application to soils (Joy et al 2013). Storage of raw manure also tends to result in degradation of antibiotics, though the occurrence of specific antibiotic resistance genes is likely related to multiple chemical and biologic stressors (Joy et al 2014) Overall these studies support continued research on the environmental occurrence, fate, and potential ecological or human health effects of these contaminants.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Jaimes-Correa, J.C., D.D. Snow and S.L. Bartelt-Hunt. 2015. Seasonal occurrence of antibiotics and a beta agonist in an agriculturally-intensive watershed. Environ Pollut 205: 87-96. doi:http://dx.doi.org/10.1016/j.envpol.2015.05.023.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Ma, R. T.C. Zhang, S. L. Bartelt-Hunt, Y. Qi, W. Kranz, D. Snow, T. Mader, C. Shapiro, D. P. Shelton, S. van Donk, D. Tarkalson, and S. Ensley. 2015. Influence of Soil Properties and Test Conditions on Sorption and Desorption of Testosterone. J. Environ. Eng., 141(7), 04015006. http://dx.doi.org/10.1061/(ASCE)EE.1943-7870.0000937
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Soni, B., S.L. Bartelt-Hunt, D.D. Snow, J.E. Gilley, B.L. Woodbury, D.B. Marx, et al. 2015. Narrow Grass Hedges Reduce Tylosin and Associated Antimicrobial Resistance Genes in Agricultural Runoff. Journal of Environmental Quality 44: 895-902. doi:10.2134/jeq2014.09.0389.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Sallach, J.B.; Zhang, Y.; Hodges, L.; Snow, D.D.; Li, X.; Bartelt-Hunt, S.L. 2015. Concomitant Uptake of Antimicrobials and Salmonella in Soil and into Lettuce Following Wastewater Irrigation. Environmental Pollution, DOI: 10.1016/j.envpol.2014.11.018


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

Outputs
Target Audience: Target audiences and efforts for this project include environmental scientists and engineers, livestock and crop producers, and regulatory agencies concerned with impact of pharmacueticals, steroid hormones and growth promoters to water quality. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? IMPACT. Row-crop and animal agriculture has changed enormously over the past century, with substantial improvements in productivity due to more efficient crop production practices, pest control and veterinary medicine. Changes to the global water cycle, however, will have critical repercussions for agriculture, food security, human health, and environmental quality.Land application of manure in agricultural watersheds is an important conservation practice under both organic and conventional food production systems. Protection of watershed water quality under increasing climate variability requires that livestock and crop producers understand best management practices to minimize water quality impacts associated with the land application of manure. These management practices can only be developed and verified by actual measurement of contaminants in manure, soils receiving manure, and run-off from crops fertilized with manure. The results of this research will permit a change in knowledge that permits prioritization and evaluation of livestock waste-borne contaminants and processes affected their fate and transport will provide for improved production techniques for crop and food animal producers and lead to practices that minimize environmental impact, sustain natural resources and ensure food safety and nutritional value. This research will also help provide the best available information to decision makers, ensure and document natural landscapes for a quality environment, as well as safe water supplies and ecosystem services for vital and vibrant rural communities. Automated methods for analysis of natural and synthetic steroids in water and solid samples were developed using liquid chromatography tandem mass spectrometry (LC-MS/MS). Atmospheric pressure photoionization (APPI) in positive mode was evaluated as an alternative to electrospray ionization (ESI) and significantly reduced background and interferences. Experiments were conducted to optimize ionization efficiency, and toluene was identified as the most appropriate dopant permitting simultaneous analysis of 20 contaminants in less than 20 minutes. Using APPI allowed the method to be optimized for both androgens and estrogens rather than having two separate ion acquisition methods as required for ESI, a significant savings in time and effort. This automated on-line method saves time and reduces interference and matrix effects using an alternative gas-phase photoionization process. A rapid and simplified method for quantitative measurement of up to 20 natural and synthetic contaminants at environmentally relevant levels in manure-impacted environments was developed and published for use in multiple projects. A variety of projects funded through several grants were used to evaluate occurrence, dissipation, treatment methods, and effect of tillage on transport of steroid hormone, pharmaceuticals and mycotoxins contained livestock manure and wastewater. One study investigated the occurrence of seventeen veterinary pharmaceuticals and thirteen steroid hormones and hormone metabolites in lagoons and adjacent groundwater at operating swine and beef cattle facilities. Another study investigated the effects of implanting, composting and tillage practices on steroid hormone fate and occurrence using a series of field and laboratory experiments comparing manure and surface run-off from cattle fed with and without the use of the growth promoters trenbolone acetate, α-zearalanol, and melengestrol acetate. Overall the highest levels of estrogen and androgen steroid hormones, ranging up to a few hundred nanograms per liter, were measured in samples of wastewater or wastewater-impacted water sources. The zearalenone-related mycotoxin compounds are frequently detected in a number of environmental samples. Estrone occurs more frequently and at higher concentrations than 17β-estradiol in surface water systems confirming previous reports by other investigators. Overall these studies indicate that naturally-produced steroid hormones and mycotoxins formed in manure and soils are much more prevalent in waste-impacted water and soil, than synthetic growth promoters use in livestock production. Composting is effective in reducing steroid hormone concentrations in livestock manure. Rainfall simulation experiments indicate that manure incorporation can reduce losses in run-off. As with surface run-off, low levels in applied manure combined with dilution and degradation will likely result in undetectable levels of hormones below the crop rooting zone. Taken together, these results indicate that transport of steroid hormones from crop land treated with beef cattle manure can be minimized through manure storage, composting, and soil incorporation practices. Interception of feedlot runoff is a key factor in minimizing potential losses to surface water environments. The co-occurrence of veterinary pharmaceutical and steroid hormone contamination in groundwater at selected livestock wastewater impoundments and the correlation between pharmaceutical occurrence in lagoon wastewater and hydraulically downgradient groundwater indicates that groundwater underlying some livestock wastewater impoundments is susceptible to contamination by veterinary pharmaceuticals and steroid hormones originating in wastewater lagoons. Parent estrogen and synthetic androgen compounds were shown to degrade rapidly to the more persistent metabolites, both of which were found predominantly associated with sediments in aquatic mesocosms.Degradation products of estrogens, androgens and mycotoxins occur in manure-impacted environments. Limited data on the occurrence of selected transformation products has been collected, in part because commercial standards are not readily available. However, trendione was identified as a stable and longer-lived transformation product in microcosm experiments. The studies supported through this research indicate that transformation products occur, can be persistent, and may have similar effects. As reference standards become available, methods can be modified to include quantification of degradation products in environmental samples.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Joy, S.R.; X. Li; D.D. Snow; J.E. Gilley, B. Woodbury and S.L. Bartelt-Hunt. Fate of Antimicrobials and Antimicrobial Resistance Genes in Simulated Swine Manure Storage. Science of the Total Environment. 481 (2014) 6974. http://dx.doi.org/10.1016/j.scitotenv.2014.02.027 Published on-line 28 February 2014
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Sangster, J. L., Y. Zhang, D. D. Snow, M.B. Cox, J.C. Sivils, Y.A. Garciac, R. Hernandezc, A.S. Kolok, and S.L. Bartelt-Hunt. The influence of sediment on steroid fate and bioavailability in aquatic systems. Science of The Total Environment, Volume 496, 15 October 2014, Pages 576-584, ISSN 0048-9697, http://dx.doi.org/10.1016/j.scitotenv.2014.07.040.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Sallach, B., Y. Zhang, L. Hodges, D. Snow, X. Li, S. Bartelt-Hunt. 2014. Accumulation of Antimicrobials in Soil and Subsequent Uptake into Lettuce Following Wastewater Irrigation. Antibiotics and Antibiotic Resistance Genes in Agroecosystems Workshop. USDA AFRI Conference Workshop, Biosphere 2. August 5-8, 2014, Oracle, AZ.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Snow, D.D, S. L. Bartelt-Hunt, S. Biswas, W. L. Kranz, T. L. Mader, C.A. Shapiro, S.J. van Donk, D. P. Shelton, T.C. Zhang, and D.D. Tarkalson. 2013 Effects of Manure Management on Steroid Hormones in the Feedlot and the Field. Special Symposium - Lessons Learned from USEPA STAR Endocrine Disrupting Chemicals Research Grants. Abstracts with Program SETAC North America 34th Annual Meeting, November 17-21, 2013, Nashville, TN.


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

Outputs
Target Audience: Target audiences and efforts for this project include environmental scientists and engineers, livestock and crop producers, and regulatory agencies concerned with impact of pharmacueticals, steroid hormones and growth promoters to water quality. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Seven gradute students in civil, environmental and biological systems engineering were supported and worked under several different field studies where samples were analyzed using these methods. Most if not all of the research publications listed in this report were co-authored by graduate students, in several cases as the lead author. While level level of analytical trained varied from project to project most students and project collborators required assistance and training in interpreting environmental data produced from these methods. Students working in these projects also presented research results through numerous conference presentations and posters listed in this report. How have the results been disseminated to communities of interest? Results of the field studies were disseminated to environmental scientists and engineers in 8 research publications including Environmental Science and Technology, Science of the Total Environment, Journal of Environmental Quality, and Transactions of ASABE. Thirteen (13) conference abstracts and proceedings used data from these advanced analytical methods in 2013. An extension publication is currently in development for producers interested in managing residues of growth promoters in cattle manure. What do you plan to do during the next reporting period to accomplish the goals? We are working on additional publications from the beef cattle manure management project detailing the effects of manure application timing on losses of steroid hormones in run-off and peristence of steroid hormones in soils fertilized with manure. Sample analyses will be interpreted for an additional project coordinated through USDA ARS Soil Agroecosystems Management unit evaluating the occurrence and treatment potential for vegatative treatment areas on removing steroids and veterinary pharmacetuicals in feedlot run-off. Finally, new research project proposals are in development and expected to support additional research using these and subsequent advanced analytical methods.

Impacts
What was accomplished under these goals? Under Objective A. our group validated and published a highly method for the determination of natural and synthetic steroids in water and solid samples were developed using liquid chromatography tandem mass spectrometry. An alternative to electrospray, atmospheric pressure photoionization (APPI), mode was used as the source which appeared to reduce background and interferences as compared to traditional ionization methods (Snow et al 2013). This method was used for several projects in analysis of field samples (Objective B). Effects of composting and soil tillage were evaluated as management tools to help control losses of hormones from beef cattle manure through a series of field and laboratory experiments comparing manure and surface run-off from cattle fed with and without the use of the growth promoters trenbolone acetate, α-zearalanol, and melengestrol acetate. Results show that wastewater or wastewater-impacted water sources (feedlot run-off, lagoon infiltration) generally show the highest levels of estrogen and androgen steroid hormones, though only ranging up to a few hundred nanograms per liter. The zearalenone-related mycotoxin compounds are frequently detected in a number of environmental samples, presumably due to in situ production in agricultural environments. Estrone generally occurs more frequently and at higher concentrations than 17β-estradiol in surface water systems as reported by other investigators. Recently deposited animal manure, and surface or groundwater directly impacted by livestock waste, generally shows the highest levels of natural steroid hormones, including synthetic growth promoters though concentrations rapidly decline over time and distance from sources. Uncontrolled runoff from confined animal production facilities can contriubte steroid hormones to the environment regardless of the use of growth promotants. Taken together, these results indicate that transport of steroid hormones from crop land treated with beef cattle manure can be minimized through manure storage, composting, and soil incorporation practices. Interception of feedlot runoff is a key factor in minimizing potential losses to surface water environments. Under Objective C, a standard for a trenbolone metabolite (trendione) was chemically synthesized, and used to confirm presence and persistence in aquatic mesocosms (Sangster et al in review). A standard for a degradation product of bacitracin A, used in swine prodcution, was synthesized and used to confirm rapid tranformation of this compound in surface runoff from soils receiving manure from swine treated with this compound and other antimicrobials (Joy et al 2013).

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Snow, D.D., T.R. Damon-Powell, S. Onanong, and D.A. Cassada. 2013. Sensitive and simplified analysis of natural and synthetic steroids in water and solids using on-line solid phase extraction and microwave-assisted solvent extraction coupled to liquid chromatography tandem mass spectrometry atmospheric pressure photoionization. Analytical and Bioanalytical Chemistry, 405, (5), 1759-1771. doi: 10.1007/s00216-012-6572-8
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Yuan, Qi; Snow, D.; Bartelt-Hunt, S. 2013. Potential water quality impacts originating from land burial of cattle carcasses. Science of the Total Environment. 456-457, 246253 http://dx.doi.org/10.1016/j.scitotenv.2013.03.083
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Bartelt-Hunt, S. L.; DeVivo, S.; Johnson, L.; Snow, D. D.; Kranz, W. L.; Mader, T. L.; Shapiro, C. A.; van Donk, S. J.; Shelton, D. P.; Tarkalson, D. D.; Zhang, T. C. 2013. Effect of Composting on the Fate of Steroids in Beef Cattle Manure. Journal of Environmental Quality, 42, (4), 1159-1166. http://dx.doi.org/10.2134/jeq2013.01.0024
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: van Donk, S. J.; S. Biswas; W. L. Kranz; D. D. Snow; S. L. Bartelt-Hunt; T. L. Mader; C. A. Shapiro; D. P. Shelton; D. D. Tarkalson; T. C. Zhang; Ensley, S., 2013. Transport of steroid hormones in the vadose zone after land application of beef cattle manure. Transactions of the ASABE, 56, (4), 1327-1338.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Biswas, S.; W. L. Kranz; C. A. Shapiro; M. Mamo; S. L. Bartelt-Hunt; D. D. Snow; D. P. Shelton; D. D. Tarkalson; T. L. Mader; S. J. van Donk; Zhang, T. C., 2013. Use of a surrogate to evaluate the impact of tillage on the transport of steroid hormones from manure-amended agricultural fields. Transactions of the ASABE, 56, (4), 1379-1385.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Zhang, Y.; Zhang, C.; Parker, D.; Snow, D.; Zhou, Z.; Li, X. 2013. Occurrence of Antimicrobials and Antimicrobial Resistance Genes in Beef Cattle Storage Ponds and Swine Lagoons. Science of the Total Environment, 463-464: 631-638. http://dx.doi.org/10.1016/j.scitotenv.2013.06.016
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Stacey, J. ; Bartelt-Hunt, S.; Snow, D. ; Gilley, J.; Woodbury, B.; Parker, D.; Marx, D.; Li, X. 2013. Fate and transport of antimicrobials and antimicrobial resistance genes in soil and runoff following land application of swine manure slurry. Environmental Science and Technology, 47(21): 12081-12088. http://dx.doi.org/10.1021/es4026358
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Joy, S.R., Bartelt-Hunt, S.L., Li, X., Snow, D.D., Gilley, J, and Marx, D. 2013. Fate and transport of chlortetracycline and resistance genes in the environment after land application of swine manure. SETAC-Europe. Glasgow, Scotland. May 12-16, 2013.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zhang, Y., D. Snow, D. Parker, Z. Zhou and X. Li. 2013. Intracellular and Extracellular Antimicrobial Resistance Genes in Livestock Manure Management Structures. 113th General meeting of American Society for Microbiology, May 18-21, 2013, Denver, CO
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zhang, Y., S. Bartelt-Hunt, D. Snow, L. Hodges, and X. Li. 2013. Internalization of Salmonella into lettuce leaves through irrigation water. 5th Quadrennial IWA Specialty Conference on Microbial Ecology and Water Engineering, Ann Arbor, MI, July 7-10.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zhang, Y., J.B. Sallach, X. Li, S. Bartelt-Hunt, D. Snow, and L. Hodges. 2013. Accumulation of antimicrobials and antimicrobial resistant Salmonella infantis in lettuce at the production stage. 68th Soil and Water Conservation Society Annual Conference, July 21-24, 2013. Reno, NV
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sallach, J.B.; Bartelt-Hunt, S.L.; Snow, D.D.; Li, X.; Hodges, L; Zhang Y. Potential for Antibiotic Uptake by Lettuce from Irrigation with Recycled Water. AEESP 50th Anniversary Conference, Golden, CO, Jul 14-16, 2013.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zhang, Y., Snow D.D., Bartelt-Hunt, S.L. (2013). Transformation of steroids in alluvial sediment. AEESP 50th Anniversary Conference, Golden, CO, Jul 14-16, 2013.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sangster, J. Snow, D.D; Bartelt-Hunt, S.L. (2013). Sorption capacity of agricultural hormones to different particle size fractions of a natural aquatic sediment. AEESP 50th Anniversary Conference, Golden, CO, Jul 14-16, 2013.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Miller, D. N. D.D. Snow, L.M. Durso, C. G. Henry, and A.M. Schmidt. 2013. Vegetative Treatment Systems to Treat Feedlot Runoff: Fate of Antibiotics and Resistant Microorganisms. Symposium--Antibiotics and Antibiotic Resistance In Agricultural Environments. ASA, CSSA, & SSSA International Annual Meetings. Nov. 3-6, 2013, Tampa FL.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Shapiro, C.A., S. Biswas, W.L. Kranz, D. P. Shelton, S. J. van Donk, D. D. Snow, S. L. Bartelt-Hunt, T. C. Zhang, T. L. Mader, D. D. Tarkalson, 2013. Effect of manure handling on steroid movement in agricultural fields from beef cattle systems. Spreading Science and Solutions. Denver, CO. April 1-5, 2013. Waste to Worth Conference, Denver CO


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

Outputs
OUTPUTS: Activities during the reporting period include collection and analysis of several hundred samples for steroid hormones, pesticides and degradation products, and pharmaceutical compounds in a variety of matrices. Several graduate and undergraduate students were trained in advanced extraction and analysis methods using both liquid chromatography tandem mass spectrometry as well as more conventional gas chromatography mass spectrometry. New and on-going collaborations using advanced detection methods include USDA Agricultural Research Services laboratories, US Department of the Interior, US Army Corp of Engineers, and researchers from Iowa State University, South Dakota State University, Cary Institute of Ecosystems Studies, and multiple investigators from the University of Nebraska. A 4-year EPA-STAR grant funded study involving development and application of an advanced method for steroid hormone analysis was completed and results reported on the EPA website at: http://cfpub.epa.gov/ncer_abstracts/index.cfm/fuseaction/display.abst ractDetail/abstract/8426 As part of this project, the results of a two-year controlled study in which beef cattle were administered steroid hormones via subcutaneous implants and feed additives used a new analytical method to demonstrate the occurrence of 16 endogenous and synthetic steroid hormones and metabolites in feedlot runoff, manure and soil. Samples were extracted and analyzed using liquid chromatography tandem mass spectrometry for metabolites of the synthetic androgen, progesterone and a wide range of endogeneous estrogens, androgens, and fusarium metabolites associated with corn. Synthetic steroids were detected in fresh manure and in feedlot surface soils from cattle administered synthetic steroids. A feed additive was detected in runoff samples, and had not been previously reported. Synthetic steroids were not detected in manure or runoff from control cattle. A wide range of endogenous hormones were detected in runoff and feedlot surface soils and manure from cattle given synthetic steroids and from control cattle, with no statistically significant differences in concentrations from treated versus untreated cattle. A PhD graduate student in Biological Systems Engineering, supported through this project, graduated and is working on additional publications to present results of 2 additional run-off studies, and a lysimeter leachate investigation using manure from the feeding pen experiments. PARTICIPANTS: Individuals who worked on this project include: Dr. Shannon Bartelt-Hunt and Dr. Tian Zhang (UNL Civil and Environmental Engineering); Dr. William Kranz, Dr. Charles Shapiro and Dr. Terry Mader (UNL Northeast Research and Extension Center and Haskell Agricultural Laboratory); and Dr. Simon van Donk (UNL West Central Research and Extension Center). Graduate students supported on this project include Sagor Biswas (Ph.D. through Biological Systems Engineering) Shannon Devivo and Rui Ma (both receiving a M.S. through the UNL Environmental Engineering program). TARGET AUDIENCES: Target audiences and efforts for this project include livestock and crop producers, environmental scientists, regulatory agencies concerned with impact of steroid hormones and implants to water quality. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The results of previous investigations had suggested livestock manure is an important source of synthetic hormones to the environment. Using advanced analytical methods, this study demonstrated that synthetic hormones used in implants occur only manure and that a synthetic progestogen occurred infrequently in feeding pen run-off. There were no significant differences in levels of naturally produced hormones in run-off from areas holding treated and untreated animals. While endogenous, naturally produced, hormones do occur in feeding pen run-off, only a small fraction of the surface run-off from feeding pens contained detectable levels of a synthetic progestagen. In addition, the formation of mycotoxins in grain, manure, and feeding pen surfaces are also an important contributor to loading of biologically active compounds through surface run-off and potentially in manure used for fertilizing crops. Additional studies investigating the fate of steroids in composted manure, and from soils fertilized with cattle manure will be described in next year's report.

Publications

  • (Journal Article) Biswas, S., C. A. Shapiro, W. L. Kranz, T. L. Mader, D. P. Shelton, D. D. Snow, S. L. Bartelt-Hunt, D. D. Tarkalson, S. J. van Donk, T. C. Zhang, S. Ensley. 2012. Current knowledge on the environmental fate, potential impact and management of growth promoting compounds used in the US beef cattle industry. Journal of Soil and Water Conservation (in Press, 9/7/2012)
  • (Journal Article) Parker, D.B; Gilley, J.; Woodbury, B. Kim, K-H.; Galvin, G. ;Bartelt-Hunt, S.L, Li, X. Snow, D.D. 2012. Odorous VOC Emission Decay Following Land Application of Swine Manure Slurry. Atmospheric Environment. doi:10.1016/j.atmosenv.2012.01.001.
  • (Journal Article) Bartelt-Hunt, S.L., D.D. Snow, W.L. Kranz, T.L. Mader, C.A. Shapiro, S.J. van Donk, D.P. Shelton, D.D. Tarkalson, and T.C. Zhang. 2012. Effect of growth promotants on the occurrence of steroid hormones on feedlot soils and in runoff from beef cattle feeding operations. Environmental Science and Technology, 46, (3), 1352-1360.
  • (Conference Abstract)Bartelt-Hunt, S.L. , D.D. Snow, W.L. Kranz, T. L. Mader, C.A. Shapiro, S.J. van Donk, D.P. Shelton, D.D. Tarkalson and T.C. Zhang. 2012. The occurrence of natural and synthetic steroid hormones on feedlot soils and in runoff from beef cattle feeding operations using growth promotants. 243rd American Chemical Society (ACS) National Meeting in San Diego, CA between March 25 and 29, 2012.
  • (Conference Abstract) Biswas, S., S.J. van Donk, W.L. Kranz, D.D. Snow, S.L. Bartelt-Hunt, T.L. Mader, C.A. Shapiro, D.P. Shelton, D.D. Tarkalson, T.C. Zhang, S. Ensley. 2012. Transport of Hormones in the Vadose Zone after Land Application of Beef Manure. American Water Resources Association. Contaminants of Emerging Concern in Water Resources. Research, Engineering, and Community Action, Denver, CO, June 25-27, 2012
  • (Conference Abstract) Mader, T. L.; L. J. Johnson, S. L. Bartelt-Hunt, D. D. Snow, W. L. Kranz, C. A. Shapiro, S. J.Van Donk, D. P. Shelton, D. D. Tarkalson, and T. C. Zhang. 2012. Fate of residual hormones from feedlot cattle. Midwest Animal Sciences Society Meeting. Des Moines, IA. March 2012.
  • (Conference Abstract) Snow, D. and S. Bartelt-Hunt. 2012. Pharmaceutical occurrence and fate in surface water using polar organic contaminant integrative samplers (POCIS) 5th International Passive Sampling Workshop (IPSW 2012) September 11-12, 2012, Columbia, MO.


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

Outputs
OUTPUTS: Several methods for organic micro-contaminants were developed during the reporting period to support water-related research. A multi-residue method combining analysis of 17 veterinary pharmaceuticals was developed and tested on surface wastewater and associated ground and surface water from several livestock facilities. These compounds were chosen based on previously developed methods and their potential for occurrence in agricultural waste. Detection limits were measured in the low ng/L (pptr) with sufficient sensitivity for environmental analysis. A second method was developed for analysis of natural and synthetic steroid hormones in livestock waste water. These compounds were chosen because of their probable occurrence in livestock waste from animals treated with growth promoting hormone. Metabolites of the synthetic hormones were included along with natural androgens and estrogens. Previous analysis of estrogenic hormones in wastewater demonstrated that severe reduction in ionization efficiency (suppression) occurs using the traditional electrospray source. Though this effect can sometimes be reduced using off-line clean up of extracts, off line clean-up is not as effective for polar (water-soluble) organic compounds such as steroids in comparison to compounds such as pesticides. Because of these difficulties atmospheric pressure photoionization (APPI) was investigated as an alternative. APPI in positive mode was found to reduced background interferences and provide more consistent ionization in complex samples. Toluene was added as a modifier (dopant) to enhance ionization efficiency. Method detection limits were determined to be in the low ng/L (pptr) range for water samples using on-line solid phase extraction. This online method, using APPI as a source and toluene as a dopant, is beneficial as it saves time due to online extraction and reduces interference and matrix effects due to the use of photoionization PARTICIPANTS: Shannon Bartelt-Hunt: University of Nebraska Civil Engineering Department. Co-investigator and collaborator. Teyona Damon-Powell: University of Nebraska School of Natural Resources graduate student developing methods used in the analysis. Ms. Damon-Powell also provided sampling assistance during the project. David Meisbach: Nebraska Department Groundwater Section Supervisor.Mr. Meisbach provided information on sampling locations and assisted with description of site hydrogeology. Nebraska Department of Environmental Quality provided funding for the field study. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
These methods were applied to calibration of passive samplers (Bartelt-Hunt et al 2011A) and to ground and surface water to investigate their occurrence and potential use as markers for waste water contamination (Bartelt-Hunt 2011B; Snow et al 2011). Several pharmaceutical and steroid hormone compounds were detected in samples obtained from animal feeding operations. Monensin and erthyromycin were consistently detected in wastewater from cattle facilities, while lincomycin and ractopamine were detected consistently in lagoon wastewater from swine facilities. Steroid hormones were identified in groundwater and lagoon samples, but were detected less frequently than veterinary pharmaceuticals. Steroid hormones detected in the wastewater samples included: estrone, testosterone, 4-androstenedione and androsterone, while only estrone and testosterone were detected in ground water samples. The levels of free hormones in wastewater samples observed in this study and the lack of detection of steroid hormones in groundwater is consistent with previous reports. Groundwater samples from the same wells showing measureable levels of individual steroid hormones or pharmaceuticals consistently produced trace residues of these compounds in subsequent samples. Results of both methods indicate that pharmaceuticals and steroid hormones can occur in livestock waste and subsequently be released to ground and surface water. Development of both methods will be described in subsequent journal articles.

Publications

  • Bartelt-Hunt, S. L.; Snow, D. D.; Damon-Powell, T.; Brown, D. L.; Prasai, G.; Schwarz, M.; Kolok, A. S. 2011A. Quantitative evaluation of laboratory uptake rates for pesticides, pharmaceuticals, and steroid hormones using POCIS. Environmental Toxicology and Chemistry 30, (6), 1412-1420.
  • Bartelt-Hunt, S.; Snow, D. D.; Damon-Powell, T.; Miesbach, D., 2011B. Occurrence of steroid hormones and antibiotics in shallow groundwater impacted by livestock waste control facilities. Journal of Contaminant Hydrology 123, (3-4), 94-103.
  • Snow, D.; D. Rus, and S. Bartelt-Hunt. 2011. Seasonal variation of surface water veterinary pharmaceutical levels in an agricultural watershed. Abstracts with Programs, EDCs and Pharmaceuticals in the Environment Session. Society of Environmental Toxicologists and Analytical Chemists. SETAC North America 32th Annual Meeting, Boston, MA.


Progress 10/01/09 to 09/30/10

Outputs
OUTPUTS: Developed and tested extraction and purification methods for steroid hormones and veterinary pharmaceuticals in water and waste water samples. using on-line solid phase extraction coupled to liquid chromatography tandem mass spectrometry. Trained graduate students in extraction techniques. Used and provided interpretation of extraction and analysis of passive samplers for pharmaceutical monitoring in a Nebraska watershed. Provided outreach to high school science class involved in water quality monitoring about the applications of passive samplers to monitor for organic contaminants in surface water. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
New automated extraction and analysis methods will be suitable for publication. Currently preparing a draft manuscript with student co-author on methods for on-line extraction and liquid chromatography tandem mass spectrometry analysis of steroid hormones in water and waste water with detection limits near 2 ng/L (pptr). New method is being used to measure natural and synthetic steroid hormones in run-off samples collected from feeding pen and rainfall simulation studies conducted to evaluate export of steroid hormones form livestock facilities and manure-fertilized fields.

Publications

  • No publications reported this period