Efficient Management and Use of Animal Manure to Protect Human Health and Environmental Quality

Goals / Objectives
The overall goal of the research project which is formulated as a real partnership between ARS and Western Kentucky University (WKU) is to conduct cost effective and problem solving research associated with animal waste management. The research will evaluate management practices and treatment strategies that protect water quality, reduce atmospheric emissions, and control pathogens at the animal production facilities, manure storage areas, and field application sites, particularly for the karst topography. This Project Plan is a unique situation in the sense that non-ARS scientists from WKU are included on an in-house project to conduct research under the NP 214. The objectives and related specific sub-objectives for the next 5 years are organized according to the Components (Nutrient, Emission, Pathogen, and Byproduct) of the NP 214, which mostly apply to this project as follows: 1) develop improved best management practices, application technologies, and decision support systems for poultry and livestock manure used in crop production; 2) develop methods to identify and quantify emissions, from poultry, dairy and swine rearing operations and manure applied lands; 3) reduce ammonia, odors, microorganisms and particulate emissions from dairy, swine and poultry operations through the use of treatment systems (e.g. biofilters and scrubbers) and innovative management practices; 4) perform runoff and leaching experiments on a variety of soils amended with dairy, swine, or poultry manures infected with Campylobacter jejuni (C. jejuni), Salmonella sp. or Mycobacterium avium subsp. paratuberculosis (MAP) and compare observed transport with that observed for common indicator organisms such as E. coli, enterococci, and Bacteriodes; and 5) use molecular-based methodologies to quantify the occurrence of pathogens and evaluate new methods to inhibit their survival and transport in soil, water, and waste treatment systems.

Project Methods
This in-house project was conceived as a cooperative/partnership and comprehensive research program between USDA-ARS Animal Waste Management Research Unit (AWMRU) and Western Kentucky University (WKU). The project is designed to utilize the scientific expertise and facilities of both institutions to conduct problem-solving research related to animal waste management in Kentucky and the southeastern U.S. The research effort will be multi-disciplinary and multifaceted in support of decision making and systems development. Research focuses will be on all four components (Nutrient, Atmospheric Emission, Pathogens, and By-products) of the National Program 214. In lieu of repeatedly stating the equipment used for analysis, please note that the state-of-the-art laboratories and equipments exist at both AWMRU and WKU, which can be accessed by the scientists including land at the WKU research station. Main instruments include: ICP, GC-MS, 2 GCs, Latchet, 2 C/N Analyzers, IC, HPLC, Real-time PCR, etc.

Progress 10/01/10 to 09/30/15

Outputs
Progress Report Objectives (from AD-416): The overall goal of the research project which is formulated as a real partnership between ARS and Western Kentucky University (WKU) is to conduct cost effective and problem solving research associated with animal waste management. The research will evaluate management practices and treatment strategies that protect water quality, reduce atmospheric emissions, and control pathogens at the animal production facilities, manure storage areas, and field application sites, particularly for the karst topography. This Project Plan is a unique situation in the sense that non-ARS scientists from WKU are included on an in-house project to conduct research under the NP 214. The objectives and related specific sub-objectives for the next 5 years are organized according to the Components (Nutrient, Emission, Pathogen, and Byproduct) of the NP 214, which mostly apply to this project as follows: 1) develop improved best management practices, application technologies, and decision support systems for poultry and livestock manure used in crop production; 2) develop methods to identify and quantify emissions, from poultry, dairy and swine rearing operations and manure applied lands; 3) reduce ammonia, odors, microorganisms and particulate emissions from dairy, swine and poultry operations through the use of treatment systems (e.g. biofilters and scrubbers) and innovative management practices; 4) perform runoff and leaching experiments on a variety of soils amended with dairy, swine, or poultry manures infected with Campylobacter jejuni (C. jejuni), Salmonella sp. or Mycobacterium avium subsp. paratuberculosis (MAP) and compare observed transport with that observed for common indicator organisms such as E. coli, enterococci, and Bacteriodes; and 5) use molecular-based methodologies to quantify the occurrence of pathogens and evaluate new methods to inhibit their survival and transport in soil, water, and waste treatment systems. Approach (from AD-416): This in-house project was conceived as a cooperative/partnership and comprehensive research program between USDA-ARS Animal Waste Management Research Unit (AWMRU) and Western Kentucky University (WKU). The project is designed to utilize the scientific expertise and facilities of both institutions to conduct problem-solving research related to animal waste management in Kentucky and the southeastern U.S. The research effort will be multi-disciplinary and multifaceted in support of decision making and systems development. Research focuses will be on all four components (Nutrient, Atmospheric Emission, Pathogens, and By-products) of the National Program 214. In lieu of repeatedly stating the equipment used for analysis, please note that the state-of-the-art laboratories and equipments exist at both AWMRU and WKU, which can be accessed by the scientists including land at the WKU research station. Main instruments include: ICP, GC-MS, 2 GCs, Latchet, 2 C/N Analyzers, IC, HPLC, Real-time PCR, etc. The research conducted under this project plan addresses several objectives/sub-objectives investigating environmental problems related to the use of animal manure and agricultural waste including nutrients, pathogens, greenhouse gases, odor causing volatile organic compounds, dust and sediment associated with animal production facilities and manure application sites. The research also determines best management practices (BMPs) for crop production on land receiving agricultural waste with regard to crop management and soil types particularly in unique �karst topography�. Following are examples of research related activities and progress for this project: Characterizing Environmental Isolates as Surrogates for Human Pathogens Associated with Animal Manures: ARS researchers in Bowling Green, Kentucky, conducted studies to identify properties of indicators that contribute to their occurrence and survival in association with produce in a manner similar to that of pathogens. ARS scientists collaborated with Western Kentucky University and characterized the adherence and growth of Escherichia coli (E. coli) isolates in greenhouse studies using fresh produce. The ARS scientist involved in this study has been asked to give a talk at the annual meeting of the Arkansas Association for Food Protection in association with this work. Aspects of this work have been presented at national and international meetings. Addressing Antibiotic Resistance (AR) Associated with Agro-ecosystems: An ARS scientist received an Organization for Economic Co-operation and Development (OECD) fellowship for a project entitled �Addressing the Global Challenge of Antibiotic Resistance: Characterizing Antibiotic Resistance Elements Associated with Agricultural Ecosystems�. This research is currently being conducted in collaboration with the Julius- Kuhn Institute in Braunschweig, Germany. The goal of the project is to evaluate mechanisms of transfer of mobile genetic elements (i.e. plasmids/ transposons) between microbial populations in poultry litters and soils with applied litters. The ARS scientist is co-organizer for a Soil Science Society of America Special Symposium funded by SSSA and Agriculture and Food Research Initiative (AFRI) entitled �Soils as the new frontier in antibiotic and antibiotic resistance discovery� and an ASA symposium entitled �Characterizing and Controlling Insects and Bacteria associated with manure-impacted environments�. This ARS scientist also works with a multi-ARS Unit initiative to organize ARS research on AR Bacteria and characterize resistance in environmental isolates of E. coli. Evaluating Transport of E. coli through Karst Environments: An ARS scientist from Bowling Green is collaborating with Western Kentucky University on a National Institute of Food and Agriculture (NIFA) funded fellowship project entitled �Mobility of 15N-tagged Escherichia coli within karst aquifers, Kentucky, USA�. As part of this project transport studies have been conducted in conjunction with natural rainfall events to evaluate the transport of Escherichia coli (E. coli) within karst watersheds in northern and southern Kentucky. These studies are field- based and will be conducted at Lexington, Kentucky, and at Crumps Cave in Bowling Green, Kentucky. Data from these studies has been presented at several conferences. Ongoing field studies using the developed analytical methods for air analyses (sub-objective 2.1): Field experiments were performed this past year at a cooperating poultry production facility in Kentucky where measured particulate matter concentrations and size distributions for the duration of a flock being raised and sold. Data analysis is ongoing. Write-ups of previous experiments performed in support of sub-objective 2. 2 continues in collaboration with the University of California, Riverside and the Claremont Colleges and two papers are in the editing phase. The focus over the time of these studies has shifted to the interaction of gas-phase nitrogen-containing compounds and sulfur-containing compounds A research proposal to the National Science Foundation was funded (to UC Riverside and ARS) and will support continuation of these collaborative studies for another three years. Research is being conducted in collaboration with researchers at North Carolina State University, University of Georgia, Oklahoma State University, University of Tennessee, Texas A&M University, Mississippi State University, and University of Florida to compare and evaluate accuracy of phosphorus indices from 10 states using water quality monitoring data collected in multiple watersheds and predictions of phosphorus (P) loss from these watersheds using fate-and-transport models. To date we have conducted model simulations for several sites in Georgia and North Carolina and have evaluated the model predictions with measured values of P loss. We have also calculated risk of P loss from several sites in Texas and Oklahoma using several phosphorus indices and have compared these risk assessments with measured P loss data. Research is being conducted in collaboration with researchers at North Carolina State University, University of Georgia, and Oklahoma State University to compare model predictions between daily and annual time step phosphorus loss models. Annual time step models are generally simpler and easier to use than daily time step models and thus are available for use by a larger audience. However, annual time step models by necessity neglect some important processes and therefore are generally viewed as less accurate than daily time step models, though direct comparisons between these models have not been conducted. This research will provide important insights into the potential differences in accuracy between daily and annual time step models and will help determine whether easy-to-use annual time step models provide reasonably good predictions of P loss compared with daily time step models to justify their use in nutrient management planning. Research investigating the effects of manure application rate and timing on transport of antibiotic resistant bacteria through soils is near completion. This research investigated whether the transport of antibiotic bacteria and their associated genes through a fine sand and a loamy sand soil is affected by the amount of manure applied and by the time interval between manure application and application of water to the soil. All experiments have been completed. Data analysis and interpretation is ongoing. This work is in support of Subobjective 4.1: Conduct transport experiments with pathogenic and indicator bacteria through soil and aquifer materials. Field study monitoring and optimization of biochemical factors on methane production from a complete mixing anaerobic digestion system (300K gallons) of poultry litter and potential co-substrates is in progress to understand the effect of operating parameters and feedstock on biogas production. Just completed, an intensive air quality monitoring study around livestock productions such as a novel high-rise- slatted-floor swine production system was carried out to understand the various characteristics of livestock air pollution for potential future adaptation of treatment system. Emissions of ammonia and greenhouse gases are being monitored from these swine production systems to see the effect of management and housing systems on the potential emissions. In the near future, these finding will be utilized and tested on a pilot-scale biofiltration systems to remove odors and greenhouse gases from livestock polluted air for use as feedstock for biofuel production. ARS scientists continued collaboration with Natural Resources Conservation Service (NRCS) scientists on a field study evaluating the impact of poultry manure, cover crop, and crop rotation on soil health and corn and soybeans yield. Also, ARS scientists are studying greenhouse gas and ammonia emissions from a new experiment on corn receiving poultry manure. Experiments with continuously fed anaerobic digesters developed to study production of biogas from poultry slaughterhouse wastewater. Valve switching systems were developed to control the flow of wastewater and gases. This is part of a dual research project to enhance production of biogas from low strength wastewater and also attempt to enhance the yield of methane from wastewater by stripping it from wastewater by using lower solubility gases. Also, digesters were constructed to investigate bubble dynamics in wastewater in an attempt to influence partitioning of biogas between the gaseous and dissolved/suspended states. Preliminary results indicate that as much of 50% of potential biogas produced by anaerobic digesters is wasted in effluent rather than being utilized. Using under water sound to destabilized suspended bubbles within the wastewater has the potential to greatly improve biogas production from anaerobic digesters. In another study, wastewater quality and biogas production from a commercial anaerobic digester utilizing poultry litter were investigated. Poultry litter digestate was characterized by high wastewater strength but low biogas quality with unusually high bicarbonate and solvated carbon dioxide concentrations. Generator startup is anticipated soon and we will be working with the producer to improve digestate utilization and in lowering carbon dioxide levels. Accomplishments 01 Effect of land application of poultry litter (PL) on survival of pathogens, indicators and genes for antibiotic resistance. PL is a valuable nutrient source for crop production, however land application without prior treatment may be a route of plant, soil or water contamination with manure-borne bacteria. In fact, two of the top bacterial causes of illness, Campylobacter sp. and Salmonella sp. are manure-borne pathogens that are found in association with poultry and PL. ARS scientists in Bowling Green, KY, in collaboration with scientists at Western Kentucky University, characterized the fate of naturally occurring pathogens, fecal indicator bacteria (FIB) and organisms containing antibiotic resistance genes (ARG) following application of PL to soils under conventional or no-till management. It was demonstrated that pathogens survive in soils with applied poultry litter in ways that are difficult to predict using common indicator organisms (i.e. E. coli) as surrogates for the manure borne pathogens. Application rates (including bacterial load and nutrients) and re- application of litter in the second year had more influence on microbial populations (particularly those with ARG) than did differences in tillage. These data provide new knowledge about survival of important FIB, pathogens, and ARG associated with PL applied under realistic field-based conditions. 02 Nitrogen source and application method impact on corn yield and nutrient uptake. Farmers are looking for better management practices to utilize animal manure as an alternative to chemical fertilizers. ARS scientists from Bowling Green, KY, conducted field experiments to study the effects of N fertilizer source and application methods to Nicholson silt loam soil in central Kentucky on no-till corn production. N fertilizer was applied as preplant and sidedress are equivalent amount of N was applied as swine effluent by three methods (broadcast, injection, and aeration) too. Results demonstrated that the swine effluent application methods and the timing of nitrogen application may not be agronomically important for corn production in this region. This research gives farmers scientifically-based information for determining the source, method, and timing of N application for corn production, thereby optimizing nutrient use for yield and environmental benefits. 03 No-till corn response to subsurface application of poultry litter. Poultry litter is generally land-applied by surface broadcast as a source of plant nutrients, particularly nitrogen (N) and phosphorus (P), for crop production. This practice is not efficient because of potential nutrient losses and environmental problems such as reduced water and air quality. ARS scientists from Bowling Green, KY, conducted a collaborative study using a new implement recently developed by ARS scientists at another location that allows subsurface application of dry poultry litter with minimal soil surface disturbance. Results indicated that poultry litter application by subsurface method resulted in corn grain and aboveground biomass yields similar to standard commercial fertilizer and better than broadcast application method. Results from this study suggest that subsurface banding of poultry litter can be utilized as an alternate application method in a no-till corn system without detrimental impacts on corn productivity. 04 Composting swine manure for stabilization and volume reduction. Over the last 30 years, animal production has become increasingly intensive with fewer operations producing larger numbers of animals. Composting swine slurries has several advantages: liquid slurries are converted to solids at lower moisture, the total volume and weight of material is reduced, and the stabilized product is more easily transported off-site. Despite this, swine waste is generally stored, treated and applied in its liquid form. High-rise finishing facilities (HRFF) permit liquid slurries to be converted to solids which are partially decomposed underneath the HRFF and then finished in compost windrows. ARS scientists from Bowling Green, KY, in collaboration with University of Kentucky Extension and a swine producer evaluated the effect of turning frequency and ambient weather conditions on biological, physical and chemical properties of composted slurry-woodchip mixtures from HRFF. Results suggest that it is feasible to finish swine HRFF materials in windrows. Volume reduction, low moisture and low readily degradable organic matter suggest that the finished compost would have lower transportation costs and should provide value as a soil conditioner. 05 Evaluation of model uncertainty using a field-scale phosphorus (P) loss model. Research was conducted to evaluate the effect of model parameter errors on model prediction uncertainties for the Annual P Loss Estimator (APLE) model, a commonly used model developed by ARS for evaluating P loss from agricultural fields. Because models are often used to predict phosphorus loss from agricultural fields, it is important to evaluate the impact that model parameter uncertainties have on model prediction uncertainties. ARS scientist from Bowling Green, KY, collaborated with other ARS scientists to improve the accuracy of the model parameter uncertainties through regression analyses using laboratory and field data. These uncertainties were incorporated into the APLE model to evaluate how these uncertainties affect overall model predictions of phosphorus loss. The relative magnitude of these uncertainties was compared to the magnitude of model input uncertainties. Results from this research demonstrate that uncertainties associated with phosphorus loss models can be relatively high and that reasonable estimates of model parameter uncertainties must be incorporated into phosphorus loss models. 06 Understanding the effect of different nutrients on biogas production from anaerobic digestion of tannery sludge wastewater. The tanning industry is one of the most highly polluted industries in the world. Tannery sludge contains recalcitrant chemicals which could make them resistant to microbial decomposition and prevent methane production. Furthermore, tannery sludge is deficient in the necessary nutrients which could affect the biogas production. Researchers from the ARS unit in Bowling Green, KY, along with scientists from Mexico, conducted experiments to understand the effect of different nutrients on biogas production from tannery sludge wastewater plant. The results showed that the addition of simple sugar (glucose), iron, and sulfate did not significantly increase the rate of biogas production. However, decreasing the carbon to nitrogen ratios by adding more nitrate and urea to the system increased the biogas production. The knowledge gained from this study could be useful for farmers and producers when carrying out anaerobic digestion of agro-industrial waste sludge to obtain biogas for power generation and maintaining sustainability. 07 Identification and characterization of microorganisms capable of reducing the toxicity of heavy metals. Heavy metal contamination poses a serious threat to both environment and human health. Thus, development of remediation strategies for heavy metals polluted soils is important for ecological conservation and health risk. A number of micro-organisms inhabiting soil and water can transform these toxic metals into a nontoxic form (biotransformation), which could reduce these toxic metals in the food chain. Researchers from ARS in Bowling Green, KY, along with scientists from the Chonbuk University in South Korea, isolated and identified microorganisms that were able to transform lead heavy metal from toxic to non-toxic form. Application of sesame oil cake extract enhanced the bacterial activity in mine drainage soils. Total lead (about 39%) was successfully transformed in mine drainage soils with sesame oil cake extract amendment. The utilization of oil cake extract not only increased the transformation rate but also improved soil quality. The present study provides a potential eco-friendly and sustained way for reducing the toxicity of lead and other heavy metal contaminants from mine drainage soils and as well as from contaminated agricultural lands.

Impacts
(N/A)

Publications

Radcliffe, D.E., Reid, D.K., Blomback, K., Bolster, C.H., Collick, A.S., Easton, Z., Francesconi, W., Fuka, D.R., Johnson, H., King, K.W., Larsbo, M., Youssef, M., Mulkey, A.S., Nelson, N.O., Persson, K., Ramirez-Avila, J. J., Schmieder, F., Smith, D.R. 2015. Applicability of models to predict phosphorus losses in drained fields: A review. Journal of Environmental Quality. 44(2):614-628.
Sharpley, A., Bergstrom, L., Aronsson, H., Bechmann, M., Bolster, C.H., Borling, K., Djodjic, F., Jarvie, H., Schoumans, O., Stamm, C., Tonderski, K., Ulen, B., Uusitalo, R., Withers, P.J. 2015. Future agriculture with minimized phosphorus losses to waters: research needs and direction. Ambio. 44:S163-S179.
Abit Jr, S.M., Bolster, C.H., Peng, H., Walker, S. 2012. Influence of feedstock and pyrolysis temperature of biochar amendments on transport of Escherichia coli in saturated and unsaturated soil. Environmental Science and Technology. 46:8097-8105.
Govarthanan, M., Cho, M., Lee, K., Park, Y., Myung, H., Krishnamoorthy, R. R., Lee, S., Lovanh, N.C., Kamala-Kannan, S., Oh, B. 2015. Lead biotransformation potential of allochthonous Bacillus sp. SKK11 with sesame oil cake in mine soil. The Royal Society of Chemistry. 5:54564- 54570.
Lovanh, N.C., Loughrin, J.H., Cook, K.L., Silva, P.J., Oh, B. 2014. Effect of windrow management on ammonia and nitrous oxide emissions from swine manure composting. International Journal of Environmental, Earth Science and Engineering. 8(6):308-312.
Lovanh, N.C., Quintana, A., Rysz, M., Loughrin, J.H., Mahmood, R. 2014. The effect of heat fluxes on ammonia emission from swine waste lagoon based on neural network analyses. Journal of Environmental Science and Technology. 7(1):16-29.
Lovanh, N.C., Loughrin, J.H., Sistani, K.R. 2012. Volatile fatty acids in suspended particulate matter from a poultry house using rice hulls as bedding materials�a profile of first flock after total cleanout. Journal Civil & Environmental Engineering. 2(4)114:1-4. doi: 10.4172/2165-784X. 1000114.
Price, D.J., Clark, C., Tang, X., Cocker, D.R., Purvis-Roberts, K.L., Silva, P.J. 2014. Proposed chemical mechanisms leading to secondary organic aerosol in the reactions of aliphatic amines with hydroxyl and nitrate radicals. Atmospheric Environment. 96:135-144.
Tang, X., Price, D.J., Praske, E., Vu, D., Purvis-Roberts, K., Silva, P.J., Cocker, D.R., Asa-Awuku, A. 2014. Cloud condensation nuclei activity of aliphatic amine secondary aerosol. Atmospheric Chemistry and Physics. 14:5959�5967.
Cook, K.L., Netthisinghe, A.M., Gilfillen, R.A. 2014. Detection of pathogens, indicators and antibiotic resistance genes following land application of poultry litter. Journal of Environmental Quality. 43:1546- 1558.
Cook, K.L., Flis, S.A., Ballard, C.S. 2013. Sensitivity of mycobacterium avium subsp paratuberculosis, escherichia coli and salmonella enterica serotype typhimurium to low pH, high organic acids and ensiling. Journal of Applied Microbiology. 115:334-345. doi:10.1111/jam.12243.
Netthisinghe, A.P., Cook, K.L., Rice, C., Gilfillen, R.A., Sistani, K.R. 2013. Soil nutrients, bacteria populations, and veterinary pharmaceuticals across a backgrounding beef feedlot. Journal of Environmental Quality. 42:532�544; DOI:10.2134.
Loughrin, J.H., Cook, K.L., Lovanh, N.C. 2012. Recirculating waste through a silicone membrane in an aerobic chamber improves biogas quality and wastewater malodors. Transactions of the ASABE. 55(5):1229-1237.
Uchimiya, M., Orlov, A., Ramakrishnan, G., Sistani, K. 2013. In situ and ex situ spectroscopic monitoring of biochar's surface functional groups. Journal of Analytical & Applied Pyrolysis. 102:53-59.
Way, T.R., Watts, D.B., Tewolde, H., Sistani, K.R., Torbert III, H.A. 2013. Implement with adjustable band spacing for subsurface band application of poultry litter. Applied Engineering in Agriculture. 29(6):831-839.
Donald, P.A., Allen, P.B., Tyler, D.D., Sistani, K.R., Tewolde, H., Walker, E.R. 2013. Effect of broiler litter application to soybean crop infested with Soybean Cyst Nematode. Nematropica. 43(1):24-34.
Jn-Baptiste, M., Sistani, K.R., Tewolde, H. 2013. Poultry litter time and method of application effects on corn yield. Soil Science. 178(3):109-119.
Tewolde, H., Sistani, K.R., Adeli, A. 2013. Fall- and spring-applied poultry litter effectiveness as corn fertilizer in the mid-southern United States. Agronomy Journal. 105:1743-1748.
Tang, X., Price, D., Praske, E.J., Lee, S., Shattuck, M.A., Purvis-Roberts, K., Silva, P.J., Asa-Awuku, A., Cocker III, D.R. 2013. NO3 and OH initiated secondary aerosol formation from aliphatic amines - salt formation and effect of water vapor. Atmospheric Environment. 72:105-112.
Netthisinghe, A., Woosley, P., Gilfillen, R., Cook, K.L., Sistani, K.R. 2014. Nutrient source and tillage impacts on tall fescue production and soil properties. Agronomy Journal. 106:1427�1437.

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

Outputs
Progress Report Objectives (from AD-416): The overall goal of the research project which is formulated as a real partnership between ARS and Western Kentucky University (WKU) is to conduct cost effective and problem solving research associated with animal waste management. The research will evaluate management practices and treatment strategies that protect water quality, reduce atmospheric emissions, and control pathogens at the animal production facilities, manure storage areas, and field application sites, particularly for the karst topography. This Project Plan is a unique situation in the sense that non-ARS scientists from WKU are included on an in-house project to conduct research under the NP 214. The objectives and related specific sub-objectives for the next 5 years are organized according to the Components (Nutrient, Emission, Pathogen, and Byproduct) of the NP 214, which mostly apply to this project as follows: 1) develop improved best management practices, application technologies, and decision support systems for poultry and livestock manure used in crop production; 2) develop methods to identify and quantify emissions, from poultry, dairy and swine rearing operations and manure applied lands; 3) reduce ammonia, odors, microorganisms and particulate emissions from dairy, swine and poultry operations through the use of treatment systems (e.g. biofilters and scrubbers) and innovative management practices; 4) perform runoff and leaching experiments on a variety of soils amended with dairy, swine, or poultry manures infected with Campylobacter jejuni (C. jejuni), Salmonella sp. or Mycobacterium avium subsp. paratuberculosis (MAP) and compare observed transport with that observed for common indicator organisms such as E. coli, enterococci, and Bacteriodes; and 5) use molecular-based methodologies to quantify the occurrence of pathogens and evaluate new methods to inhibit their survival and transport in soil, water, and waste treatment systems. Approach (from AD-416): This in-house project was conceived as a cooperative/partnership and comprehensive research program between USDA-ARS Animal Waste Management Research Unit (AWMRU) and Western Kentucky University (WKU). The project is designed to utilize the scientific expertise and facilities of both institutions to conduct problem-solving research related to animal waste management in Kentucky and the southeastern U.S. The research effort will be multi-disciplinary and multifaceted in support of decision making and systems development. Research focuses will be on all four components (Nutrient, Atmospheric Emission, Pathogens, and By-products) of the National Program 214. In lieu of repeatedly stating the equipment used for analysis, please note that the state-of-the-art laboratories and equipments exist at both AWMRU and WKU, which can be accessed by the scientists including land at the WKU research station. Main instruments include: ICP, GC-MS, 2 GCs, Latchet, 2 C/N Analyzers, IC, HPLC, Real-time PCR, etc. This project plan under several objectives/sub-objectives investigates environmental problems related to the improper use of animal manure and agricultural waste including nutrients, pathogens, greenhouse gases (GHGs) , odor-causing volatile organic compounds, dust and sediment associated with animal production facilities and manure application sites. Research also determines best management practices (BMPs) for crop production on land receiving agricultural waste with regard to crop management and soil types particularly in unique �karst topography�. Following are research related activities for the past year: In collaboration with scientists at Western Kentucky University (WKU), ARS researchers in Bowling Green, KY, completed a two year field study to evaluate the survival of naturally occurring pathogens (Campylobacter jejuni, Listeria monocytogenes and Salmonella enterica), indicator organisms (Escherichia coli (E. coli), enterococci) and antibiotic resistance (AR) genes in applied poultry litter. Data have been presented at three meetings and two manuscripts. An ARS scientist is serving as primary mentor for a female University of Kentucky PhD student on a NIFA funded fellowship project. She is working on research to evaluate the transport of Escherichia coli (E. coli) within karst watersheds in northern and southern Kentucky. These studies are field-based and will be conducted at Lexington and at Crumps Cave in Bowling Green, KY. As part of a two year research grant funded by the Center for Produce Safety, ARS researchers in Bowling Green, KY, are conducting studies to identify properties of indicators that contribute to their occurrence and survival in association with produce in a manner similar to that of pathogens. As part of the study, a graduate student from WKU is working at the ARS site to characterize the adherence and growth of Escherichia coli (E. coli) isolates on lettuce contaminated from manure application or irrigation. Also, ARS researchers in Bowling Green, KY, are working in collaboration with scientists at the University of Iowa and ARS USDA-ARS in Ames, IA, to characterize microbial populations (i.e., methanogens and sulfate reducers) associated with foaming in swine pits. ARS scientists continued collaboration with Natural Resources Conservation Service scientists on a field study evaluating the impact of poultry manure, cover crop, and crop rotation on soil health and corn and soybeans yield. Also, ARS scientists are studying greenhouse gas and ammonia emissions from a new experiment on corn receiving poultry manure. Research funded by the National Pork Board is being conducted to investigate the effects of manure application rate and timing on transport of antibiotic resistant bacteria through soils. This research is investigating whether the transport of antibiotic bacteria and their associated genes through a fine sand and a loamy sand soil is affected by the amount of manure applied and by the time interval between manure application and application of water to the soil. Experiments using a fine sand have been completed and the data are currently being analyzed. Experiments with loamy soil are just beginning. Research is being conducted in collaboration with researchers at North Carolina State University, University of Georgia, Oklahoma State University, University of Tennessee, Texas A&M University, Mississippi State University, and University of Florida to compare and evaluate accuracy of phosphorus (P) indices from 10 states using water quality monitoring data collected in multiple watersheds and predictions of P loss from these watersheds using fate-and-transport models. To date ARS Researchers have conducted model simulations for several sites in Georgia and North Carolina and have evaluated the model predictions with measured values of P loss. ARS Researchers have also calculated the risk of P loss from several sites in Texas and Oklahoma using several P indices and have compared these risk assessments with measured phosphorus loss data. Research is conducted to evaluate the effect of model parameter uncertainties for assessing uncertainties in the Annual Phosphorus (P) Loss Estimator (APLE) model, a commonly used model developed by ARS for evaluating P loss from agricultural fields. Because models are often used to predict P loss from agricultural fields, it is important to evaluate the impact that model parameter uncertainties have on model prediction uncertainties. To date work has focused on estimating uncertainties for model parameters obtained by regression with observed laboratory and field data. These uncertainties will be incorporated into the APLE model to evaluate how these uncertainties affect overall model predictions of P loss. Continuously fed Anaerobic Digesters were developed to study production of biogas from poultry slaughterhouse wastewater. Valve switching systems were developed to control the flow of wastewater and gases. This is part of a dual research project to enhance production of biogas from low strength wastewater and also attempt to enhance the yield of methane from wastewater by stripping it from wastewater by using lower solubility gases. Further research was conducted for improving the isolation and quantification of tetracycline antibiotics from animal wastes. After accepted methods for the isolation of these antibiotics from environmental samples proved inadequate, new methods employing a combination of reverse phase and anionic separation on solid phase sorbents were developed by discussion with commercial vendors and colleagues at Western Kentucky University. Methods for the quantification of hydrogen sulfide from biogas were developed. Calibration curves were developed for hydrogen sulfide and other malodorous and toxic sulfur compounds and dilutions for biogas and wastewater samples were determined to account for low linearity response of flame photometric detector gas chromatograph. Significant Activities that Support Special Target Populations: Two ARS scientists are serving on committee of one female PhD graduate student from the University of Kentucky and one minority female PhD student from Alabama A&M University. Served on Soil science society Women in Agriculture Scholarship committee enabling underrepresented groups to attend the tri-societies meetings and ensuring quality representatives and research are part of the agricultural and soil science meetings; serving soil and agricultural scientists and under-represented groups. Accomplishments 01 Mapping the spatial distribution of manure contaminants around livestock operations. Working in collaboration with ARS scientists in Beltsville, MD, and scientists at Western Kentucky University, ARS scientists in Bowling Green, KY, completed a study to evaluate the spatial distribution of bacteria, nutrients and veterinary pharmaceuticals across a beef cattle backgrounding operation. Intensive management of livestock may increase the potential for release of manure contaminants into soils and water sources. To determine how contaminants are transported through the environment and to pinpoint contaminant �hotspots� researchers used spatial sampling and mapping. Results showed that there was no distinct downhill flow pattern. Instead, contaminants were localized around the barn, suggesting that cleanup efforts focused on the dirt and feeder areas of the barn may go a long way in alleviating risks associated with the manure. 02 Corn yield and nutrient uptake response to swine manure application methods. Farmers are looking for better management practices to enhance production and reduce negative environmental impact from nitrogen (N) fertilizer application since N is one of the most important and costly nutrient inputs for crop production. In this field experiment three pre-plant swine effluent application methods (surface broadcast, direct injection, and application in combination with soil aeration �Aeration method�) were evaluated for no-till corn grain production. Corn grain yield was not significantly different among the three methods of swine effluent applications. However, the injection method produced the greatest corn grain yield. Results demonstrated that the swine effluent and fertilizer application method may not be very important agronomically for corn production in this region, but injecting effluent and N fertilizer may prevent nutrient losses and negative environmental impact when evaluating the impact of N fertilization and liquid manure management. 03 Bacterial transport through biochar-amended soils. ARS researchers located in Bowling Green, Kentucky, investigated the role of soil texture and bacterial surface properties on the transport behavior of two pathogenic bacteria through two biochar-amended soils. Results from the transport studies and auxiliary batch studies indicate that changes in cell retention following biochar amendments were likely due to changes in bacterial attachment in the column. Research also indicated that changes in bacterial hydrophobicity following biochar amendments generally coincided with changes in bacterial retention. The influence of biochar amendment in increasing retention of both bacteria was generally more pronounced in fine sand and indicates that soil texture does indeed affect the transport behavior of bacteria through biochar- amended soils. Incorporation of biochar into soils has been proposed as a means to sequester carbon from the atmosphere and to increase soil retention of agrochemicals. This research demonstrates the potential use of biochar as a means of reducing pathogen leaching into groundwater and provides some insights into the mechanisms involved. 04 Adaptation of a sampling trailer for online, in-situ air quality measurements. ARS researchers at the Food Animal Environmental Systems Research Unit (FAESRU) in Bowling Green, Kentucky, completed adaptation of a trailer to allow for state-of-the-art sampling of gases and particulates in field work. The air sampling trailer contains a suite of instrumentation unique within the agency for detecting and quantifying primary and secondary aerosol emissions in the field. Measurement technology available for field work with the FAESRU air quality trailer include: (1) Three minute measurements of particle concentration from 10�500 nanometers, (2) Twenty second measurements of particle concentration from 0.3 - 20 micrometers, (3) Thirty minute measurements of gas and particulate ammonium, chloride, nitrate, sulfate, amines, and carboxylic acids, and (4) Ten minute gas measurements of hydrogen sulfide, methanethiol, dimethylsulfide, and larger organic sulfur compounds. 05 Understanding the effect of heat fluxes on ammonia (NH3) and greenhouse gas emissions from swine waste lagoon. Anaerobic lagoons are effective and low-cost bioreactors to treat animal manure but they are also responsible for emissions of numerous atmospheric pollutants including NH3, greenhouse gases, and odorous compounds such as skatoles. Researchers from the ARS unit in Bowling Green, KY, along with researchers from Western Kentucky University and the University of Iowa conducted experiments to understand the effect of heat fluxes on the emission of ammonia and greenhouse gases from swine waste lagoons. They examined the effect of photochemical processes such as solar radiation on ammonia and greenhouse gases emission profile from an anaerobic swine lagoon as a case study for adopting better management strategies and designing alternative remedial options. Mathematical models based on artificial neural network were constructed to simulate and understand the effect of photochemical processes on the emissions of ammonia and greenhouse gases from swine waste lagoon. These models could serve as useful tools for evaluating and implementing best management practices in minimizing ammonia and greenhouse gases emissions from swine or any other livestock waste receptacles. Incorporating various heat fluxes such as net solar radiation, sensible heat, and latent heat of vaporization improves the accuracy of the predictive model on ammonia and greenhouse gases emissions. 06 Investigating the effect of windrow management on ammonia and greenhouse gases emissions from swine manure composting. The method of application of liquid manure is the surface spray, in which the manure is broadcast-applied on the soil surface. This method can lead to major losses of essential nutrients for crops such as nitrogen and carbon compounds. This technique can also create a major emission problem in dispersing malodorous and other gaseous compounds in the air. Composting of these animal manures is a viable option for biomass and pathogenic reduction in the environment. Nevertheless, composting also increases the potential loss of available nutrients for crop production as well as unwanted emission of anthropogenic air pollutants due to the loss of ammonia and other compounds via volatilization. Researchers from the ARS unit in Bowling Green, KY, conducted experiments to understand the effect of windrow management on the emissions of ammonia and greenhouse gases during the composting of swine manure amended with woodchip as absorbent materials. Results showed that turning frequency of compost piles not only physically increased the emissions of ammonia and greenhouse gases, but also have biochemical effect on the production of these air pollutants in the compost piles. Although composting of animal waste is quite beneficial for biomass reduction, composting may not be economically feasible from an agronomical point of view due to time, nutrient loss (N loss), and potential environmental pollution (ammonia and greenhouse gas emissions). This information could be useful for farmers and producers when carrying out composting of liquid animal manures.

Impacts
(N/A)

Publications

Loughrin, J.H., Lovanh, N.C., Cook, K.L. 2013. Improvement of anaerobic digester performance by wastewater recirculation through an aerated membrane. Transactions of the ASABE. 56(5):1675-1681.
Abit, S.M., Bolster, C.H., Cantrell, K.B. 2014. Transport of Escherichia coli O157:H7 and Salmonella typhimurium in biochar-amended soils with different textures. Journal of Environmental Quality. 43:371-378.
Sistani, K.R., Jn-Baptiste, M., Simmons, J.R. 2014. Corn response to enhanced-efficiency nitrogen fertilizers and poultry litter. Agronomy Journal. 106(2):761-770.
Lanphere, J.D., Rogers, B., Luth, C., Bolster, C.H., Walker, S.L. 2014. Stability and transport of graphene oxide nanoparticles in groundwater and surface water. Environmental Engineering Science. 31(5):1-10.
Quintanar, A., Mahmood, R., Lovanh, N.C., Rawley, J.M., Becerra-Acosta, E., Loughrin, J.H. 2013. Estimating greenhouse gas emissions from a waste lagoon. Applied Engineering in Agriculture. 29(4):511-519.

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

Outputs
Progress Report Objectives (from AD-416): The overall goal of the research project which is formulated as a real partnership between ARS and Western Kentucky University (WKU) is to conduct cost effective and problem solving research associated with animal waste management. The research will evaluate management practices and treatment strategies that protect water quality, reduce atmospheric emissions, and control pathogens at the animal production facilities, manure storage areas, and field application sites, particularly for the karst topography. This Project Plan is a unique situation in the sense that non-ARS scientists from WKU are included on an in-house project to conduct research under the NP 214. The objectives and related specific sub-objectives for the next 5 years are organized according to the Components (Nutrient, Emission, Pathogen, and Byproduct) of the NP 214, which mostly apply to this project as follows: 1) develop improved best management practices, application technologies, and decision support systems for poultry and livestock manure used in crop production; 2) develop methods to identify and quantify emissions, from poultry, dairy and swine rearing operations and manure applied lands; 3) reduce ammonia, odors, microorganisms and particulate emissions from dairy, swine and poultry operations through the use of treatment systems (e.g. biofilters and scrubbers) and innovative management practices; 4) perform runoff and leaching experiments on a variety of soils amended with dairy, swine, or poultry manures infected with Campylobacter jejuni (C. jejuni), Salmonella sp. or Mycobacterium avium subsp. paratuberculosis (MAP) and compare observed transport with that observed for common indicator organisms such as E. coli, enterococci, and Bacteriodes; and 5) use molecular-based methodologies to quantify the occurrence of pathogens and evaluate new methods to inhibit their survival and transport in soil, water, and waste treatment systems. Approach (from AD-416): This in-house project was conceived as a cooperative/partnership and comprehensive research program between USDA-ARS Animal Waste Management Research Unit (AWMRU) and Western Kentucky University (WKU). The project is designed to utilize the scientific expertise and facilities of both institutions to conduct problem-solving research related to animal waste management in Kentucky and the southeastern U.S. The research effort will be multi-disciplinary and multifaceted in support of decision making and systems development. Research focuses will be on all four components (Nutrient, Atmospheric Emission, Pathogens, and By-products) of the National Program 214. In lieu of repeatedly stating the equipment used for analysis, please note that the state-of-the-art laboratories and equipments exist at both AWMRU and WKU, which can be accessed by the scientists including land at the WKU research station. Main instruments include: ICP, GC-MS, 2 GCs, Latchet, 2 C/N Analyzers, IC, HPLC, Real-time PCR, etc. This project plan under several objectives/sub-objectives investigates environmental problems related to the improper use of animal manure and agricultural waste in relation to nutrients, pathogens, greenhouse gases (GHGs), odor-causing compounds, dust and sediment associated with animal production facilities and manure application sites. Research also determines best management practices (BMPs) for crop production on land receiving agricultural waste with regard to crop management and soil types particularly in this unique �karst topography� region. Following are research related activities for the past year: Completed the 2nd year of a study to evaluate the survival of naturally occurring pathogens (Campylobacter jejuni, Listeria monocytogenes and Salmonella enterica) and indicator organisms (Escherichia coli [E. coli], enterococci) in applied poultry litter or dairy manure. A new collaboration was established with the University of Kentucky evaluating the transport of E. coli strains with different adhesion properties through karst systems. The National Pork Board Research has provided funding to investigate the effects of manure application rate and timing on transport of antibiotic resistant bacteria through soils. Gas quality and dissolved gases in anaerobic digesters are being studied. Concentrations of methane and carbon dioxide in the biogas produced by anaerobic digestion of swine waste and other agricultural wastes are quantified in conjunction with those of dissolved methane, carbon dioxide and bicarbonate in wastewater. Initiated a study to quantify antibiotics in swine manure. Four antibiotics (penicillin, tetracycline, chlortetracycline and oxytetracycline) that are commonly added to swine feed to prevent disease and promote weight gain are being studied to determine their concentrations at all stages of swine production from birth to slaughter. Also, ARS scientists at Bowling Green, KY, initiated collaboration with ARS scientists in Lincoln, NE, to optimize methods and procedures for extracting and amplifying antibiotic resistant (AR) bacteria and their genes in agricultural matrices. An ARS scientist at Bowling Green, KY, has developed assays to target 5 different groups of AR genes (tetracyclines, sulfonamides, erythromycin, ampicillin and streptomycins). ARS scientists in Bowling Green deployed the ambient ion monitor method (AIM) to detect gas and particle phase amines and gas phase sulfur compounds at a swine facility in Kentucky. Also, Smog chamber experiments on amines were conducted in collaboration with the University of California at Riverside. The swine manure composting study is still underway; also measuring the ammonia and greenhouse gases from swine manure compost systems to see the effect of management on the potential emissions. Research is being conducted in collaboration with researchers at eight major universities to compare and evaluate accuracy of phosphorus indices from 10 states using water quality monitoring data collected in multiple watersheds using and predictions of P loss from these watersheds using fate-and-transport models. Significant Activities that Support Special Target Populations: Fact Sheet �Amending poultry litter to reduce ammonia producing bacteria� targeted to farmers, integrators and U.S. Poultry & Egg association members. Fact Sheet �Johne�s Disease: Troughs as a hidden source� targeted to dairy farmers and members of the International Association for Paratuberculosis. Scientist served on panel to review grant proposal submitted to USDA- NIFA. Scientist reviewed grant proposal submitted to the Wisconsin Water Resources Research Institute. Scientist was asked by the Kentucky Agricultural Water Quality Authority to provide comments on proposed revisions to their Nutrient Management Planning guidelines. Scientist served on 3 committees for two scientific organizations (Soil Science Society of America and American Society of Agronomy). Served on committee of one female graduate student who completed MS degree at Western Kentucky University Department of Agriculture. Presented talk to 20 Western Kentucky University Students on ARS and role of women in agricultural research; Transfer information about importance of women in agricultural research and overview of research being conducted by AWMRU Served on Soil Science Society Women in Agriculture Scholarship committee enabling underrepresented groups to attend the tri-societies meetings and ensuring quality representatives and research are part of the agricultural and soil science meetings; serving soil and agricultural scientists and under-represented groups. Accomplishments 01 Use of biochar to increase Escherichia coli (E. coli) retention in soils. The land application of animal and human fecal material poses a potential public health risk if humans become exposed to fecal-borne pathogenic microorganisms such as bacteria, viruses, and protozoa. One potential route of transmission for these pathogenic microorganisms is through drinking fecally-contaminated groundwater. One management practice which has the potential for reducing contamination of groundwater supplies by pathogenic microorganisms is amending soils with charred organic matter derived from plant biomass or bio-waste. This material, termed biochar, has been proposed as a means to sequester carbon from the atmosphere. ARS scientists from Bowling Green, KY, in collaboration with the University of California at Riverside conducted experiments to compare the effect of different types of biochar on the surface properties and transport behavior Escherichia coli (E. coli) through soil columns. Results showed that the source of biochar can have a strong impact on the mobility of E. coli through soil. Specifically, biochar produced from pine chips was found to be much more effective in restricting E. coli transport than biochar produced from poultry litter. 02 Pathogens responsible for Johne�s disease and gastroenteritis are surprisingly resistant to ensiling process. Livestock production relies heavily on silage materials as feed during winter months. The low potential of hydrogen (pH) and high organic acid concentrations produced during ensiling result in a well-preserved highly nutritious feedstock. However, organisms that exhibit acid tolerance may survive the ensiling process and can be a concern for food and animal safety when contaminated materials are fed to livestock. Researchers at the ARS in Bowling Green, KY, and from the Miner Agricultural Research Institute in Chazy, NY, evaluated the susceptibility of two acid tolerant pathogens to silage conditions. Mycobacterium avium subsp paratuberculosis (M. paratuberculosis) causes Johne�s disease in cattle and Salmonella enterica serotype Typhimurium (S. Typhimurium) is a common human gastrointestinal pathogen. Researchers found that while S. Typhimurium was relatively sensitive to ensiling M. paratuberculosis was especially resistant, surviving longer and in higher concentrations. These results suggest that when present in manure and applied to forage grasses that are ensiled, M. paratuberculosis may survive the ensilaging process and could, therefore, be a potential route of infection if ingested by a susceptible animal. 03 Antibiotic resistance (AR) genes, pathogens and indicator organisms in land applied manures. ARS researchers in Bowling Green, KY, working in collaboration with scientists at Western Kentucky University applied poultry litter and dairy manure to forage grasses under conventional and no till management. They found that even when gastrointestinal pathogens like Campylobacter jejuni and Salmonella enterica were present in manures at high concentrations initially, they were rarely detectable following land application. Indicator organisms like enterococci were measurable but the common indicator Escherichia coli (E. coli) was not. They found that Antibiotic resistance (AR) genes were very high in poultry litter before application and concentrations remained high following land application of litter. Detection of AR genes, pathogens and indicators following manure application was irregular over the sampling period suggesting that factors such as moisture and temperature play an important role in persistence in soil. This information should be considered in any risk assessment model and is important for stakeholders using livestock manures for land application onto forage grasses. 04 Improvements in wastewater biogas production. Methane is produced by the breakdown of animal manures in the absence of oxygen. This methane remains a largely untapped source of energy due to the corrosive nature of the gas. Hydrogen sulfide and carbon dioxide are the major contaminants of the natural gas produced by the microbial breakdown of animal wastes. These compounds reduce the heating value of the gas and lead to costly maintenance problems. ARS researchers in Bowling Green, KY, circulated wastewater from an anaerobic digester through a silicone hose located in an external tank. The tank was filled with a buffer solution designed to remove carbon dioxide from the wastewater. This treatment reduced carbon dioxide concentrations in the biogas resulting in a fuel with higher heating value. Improved processes for the production of biogas can lead to significant energy savings for producers and industry as well as reduced greenhouse gas emissions to the environment. 05 Development of a new phosphorus (P) index for Kentucky. In most states, the phosphorus (P) index is the adopted strategy for assessing a field�s vulnerability to P loss as required by the USDA-NRCS 590 Nutrient Management Standard. Recently, USDA-NRCS revised its 590 Standard to require that states demonstrate the accuracy of their P index. Previous research by ARS researchers in Bowling Green, KY, identified several important limitations with the existing P index for Kentucky (KY). As a result, ARS was asked to lead an effort to develop a new P index for KY with representatives from the University of KY, KY USDA-NRCS, KY Division of Water, and KY Division of Conservation. A new P index was developed to reflect current understandings of the processes controlling P loss at the field scale. The new P index was evaluated against P runoff data reported in the literature and was shown to provide significantly better predictions of P loss from agricultural fields than the original KY P index. The new index provides users with a more accurate tool for assessing the risk of P loss from agricultural fields. The new P index has been adopted by Kentucky USDA-NRCS as part of their 590 Nutrient Management Standard. 06 Developing methods for evaluating the accuracy of a phosphorus (P) index. A phosphorus index (PI) is a very simple tool used by most states to evaluate the risk of phosphorus (P) leaving agricultural fields and entering streams and lakes where it may cause water-quality deterioration. While the PI is a commonly used tool, most state P indices have not been rigorously evaluated against measured P loss data to determine how well the PI assigns P loss risk � a major reason being the lack of field data available for such an analysis. Given the lack of P loss data available for PI evaluation, ARS scientists from Bowling Green, KY, and other units demonstrated a method for evaluating P indices using output from more complex P loss models. Applying this approach to an existing state P index, ARS researchers from Bowling Green, KY, were able to identify several potential limitations with a commonly used formulation of the P index. Moreover, results from this research showed how this new approach can be used to improve how each factor in the P index is weighted. Results from this research have been used to inform developers modifying P indices in Kentucky, Maryland, Oregon, and Washington. 07 Incorporating uncertainties in predictions from phosphorus loss models. Models are often used to predict phosphorus (P) loss from agricultural fields. While it is commonly recognized that model predictions are inherently uncertain, few studies have addressed prediction uncertainties using P loss models. Research conducted by ARS scientists in Bowling Green, KY, evaluated two methods for assessing uncertainties in the Annual P Loss Estimator (APLE) model, a commonly used model developed by ARS for evaluating P loss from agricultural fields. This research resulted in identifying a simple method for calculating prediction uncertainties for the APLE model. The research also demonstrated the range in magnitude of model prediction uncertainties associated with uncertainties in model input variables. In particular, results showed that prediction uncertainties due to errors in model inputs can range from � 2 to 64% of the predicted value. Because the APLE model has been used to evaluate the accuracy of P loss models and to identify practices which help minimize P loss from agricultural fields, implementation of uncertainty to model predictions will provide improved estimates of P loss when using this model. 08 Corn Response to Enhanced-Efficiency Nitrogen Fertilizers (EENF) and Poultry Litter. Nitrogen (N) is one of the most important nutrients and costly input for crop production. Farmers are looking for better management practice to enhance N utilization and crop production while reducing environmental impact. ARS scientists in Bowling Green, KY, have investigated corn (Zea mays L.) grain yield and nutrient uptake resulting from application of 168 kg N ha-1 of several inorganic N fertilizers, commercially available Enhanced-Efficiency Nitrogen Fertilizers (EENF), and poultry litter under no-till corn production. All N sources increased corn grain yield over control (no N applied), however, no significant differences were observed among the EENF and other N fertilizer sources. This research will help farmers with needed scientific information, because lack of research-based recommendations as to when and where these EENF would be most effective, and which combination of management practices would optimize their performances. 09 Association of odors (volatile fatty acids) in suspended particulate matters from poultry house air emissions. As more and more animal production continues to evolve into major concentrated operations, odor emissions from these confined animal feeding operations (CAFOs) have caused persistent public concern and complaints. In addition, particulate matters and other gas emissions have also been a cause for environmental and health concerns. Researchers from ARS in Bowling Green, KY, conducted experiments to understand the association of odors in suspended particulate matters from poultry houses to examine and characterize the profile of various volatile fatty acids in the inhalable fractions of suspended particulate matters from a poultry house that used rice hulls as bedding materials over a period of one flock. The results showed that propionic and butyric acid are the major volatile fatty acids found in the inhalable fraction of dust collected from the broiler house after a total cleanout. Trace amounts of other volatile fatty acids (i.e., acetic acid, pentanoic acid, and hexanoic acid) were also observed from particulate matter analyses. Based on these data, particulate matters could serve as potential carriers for short and long-range transport of odorous compounds such as volatile fatty acids.

Impacts
(N/A)

Publications

Bolster, C.H., Vadas, P.A., Sharpley, A.N., Lory, J.A. 2012. Using a P loss model to evaluate and improve P indexes. Journal of Environmental Quality. 41:1758-1766.
Sharpley, A., Beegle, D., Bolster, C.H., Good, L., Joern, B., Ketterings, Q., Lory, J., Mikkelsen, R., Osmond, D., Vadas, P.A. 2012. Phosphorus indices: Why we need to take stock of how we are doing. Journal of Environmental Quality. 41(6):1711-1719.
Osmond, D., Cabrera, M., Sharpley, A., Bolster, C.H., Feagley, S., Lee, B., Mitchell, C., Mylavarapu, R., Oldham, L., Walker, F., Zhang, H. 2012. Comparing southern P indices to runoff data. Journal of Environmental Quality. 41:1741-1749.
Cook, K.L., Flis, S.A., Ballard, C.S. 2013. Mycobacterium avium subsp paratuberculosis cells are surprisingly resistant to ensiling process. Paratuberculosis Newsletter. March 2013 p7-8.
Erupe, M.E., Liberman-Martin, A., Silva, P.J., Malloy, Q.G., Yonis, Y., Cocker, III, D.R., Purvis-Roberts, K. 2010. Determination of Methylamines and Trimethylamine-N-oxide in particulate matter by non-suppressed ion chromatography. Journal of Chromatography. 1217(13):2070-2073.
Loughrin, J.H., Bolster, C.H., Lovanh, N.C., Sistani, K.R. 2010. A simple device for the collection of water and dissolved gases at defined depths. Applied Engineering in Agriculture. 26(4):559-564.
Quintanar, A.I., Mahmood, R., Loughrin, J.H., Lovanh, N.C., Motley, M.V. 2009. A system for estimating bowen ratio And evaporation from waste lagoons. Applied Engineering in Agriculture. 25(6):923-932.
Loughrin, J.H., Quintanar, A., Mahmood, R., Lovanh, N.C. 2010. Heat flux measurements and modeling of malodorous compounds above an anaerobic swine lagoon. Water, Air, and Soil Pollution. 217(1-4):463-471.
Sistani, K.R., Mikha, M.M., Warren, J.G., Gilfillen, B., Acosta Martinez, V. 2011. Nutrient source and tillage impact on corn grain yield and soil properties. Soil Science. 175(12):593-600.
Gilfillen, R.A., Rowland, N.S., Willian, W.T., Sleugh, B.B., Tekeste, M.Z., Sistani, K.R. 2010. Effects of broiler litter application on nutrient accumulations in soil.. Forage and Grazinglands. doi:10.1094/FG-2010-1105- 01-RS.
Jn-Baptiste, M., Sistani, K.R., Tewolde, H. 2012. Poultry manure application time impact on corn grain production in a crider silt loam. Soil Science. 177(1):47-55.
Gilfillen, B., Rowland, N., Willian, T., Sleugh, B.B., Tekeste, M.Z., Sistani, K.R. 2010. Effects of broiler litter application on nutrient accumulation in soil. Forage and Grazinglands. On Line Only- doi:10.1094/ FG-2010-1105-01-RS.
Bolster, C.H. 2011. A critical evaluation of the Kentucky phosphorus index. Journal of the Kentucky Academy of Science. 72(1):46�58.
Loughrin, J.H., Quintanar, A.I., Cook, K.L., Lovanh, N.C., Mahmood, R., Bexerra-Acosta, E. 2012. Seasonal variation in heat fluxes, predicted emissions of malodorants, and wastewater quality of an anaerobic swine waste lagoon. Water, Air, and Soil Pollution. 223:3611�3618.
Cook, K.L., Bolster, C.H., Ayers, K.A., Reynolds, D.N. 2011. Escherichia coli diversity in livestock manures and agriculturally impacted stream waters. Current Microbiology. 63(5):439�449.

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

Outputs
Progress Report Objectives (from AD-416): The overall goal of the research project which is formulated as a real partnership between ARS and Western Kentucky University (WKU) is to conduct cost effective and problem solving research associated with animal waste management. The research will evaluate management practices and treatment strategies that protect water quality, reduce atmospheric emissions, and control pathogens at the animal production facilities, manure storage areas, and field application sites, particularly for the karst topography. This Project Plan is a unique situation in the sense that non-ARS scientists from WKU are included on an in-house project to conduct research under the NP 214. The objectives and related specific sub-objectives for the next 5 years are organized according to the Components (Nutrient, Emission, Pathogen, and Byproduct) of the NP 214, which mostly apply to this project as follows: 1) develop improved best management practices, application technologies, and decision support systems for poultry and livestock manure used in crop production; 2) develop methods to identify and quantify emissions, from poultry, dairy and swine rearing operations and manure applied lands; 3) reduce ammonia, odors, microorganisms and particulate emissions from dairy, swine and poultry operations through the use of treatment systems (e.g. biofilters and scrubbers) and innovative management practices; 4) perform runoff and leaching experiments on a variety of soils amended with dairy, swine, or poultry manures infected with Campylobacter jejuni (C. jejuni), Salmonella sp. or Mycobacterium avium subsp. paratuberculosis (MAP) and compare observed transport with that observed for common indicator organisms such as E. coli, enterococci, and Bacteriodes; and 5) use molecular-based methodologies to quantify the occurrence of pathogens and evaluate new methods to inhibit their survival and transport in soil, water, and waste treatment systems. Approach (from AD-416): This in-house project was conceived as a cooperative/partnership and comprehensive research program between USDA-ARS Animal Waste Management Research Unit (AWMRU) and Western Kentucky University (WKU). The project is designed to utilize the scientific expertise and facilities of both institutions to conduct problem-solving research related to animal waste management in Kentucky and the southeastern U.S. The research effort will be multi-disciplinary and multifaceted in support of decision making and systems development. Research focuses will be on all four components (Nutrient, Atmospheric Emission, Pathogens, and By-products) of the National Program 214. In lieu of repeatedly stating the equipment used for analysis, please note that the state-of-the-art laboratories and equipments exist at both AWMRU and WKU, which can be accessed by the scientists including land at the WKU research station. Main instruments include: ICP, GC-MS, 2 GCs, Latchet, 2 C/N Analyzers, IC, HPLC, Real-time PCR, etc. Research related to this project plan under several objectives/sub- objectives investigates environmental problems related to the improper use of animal manure and agricultural waste including nutrients, pathogens, greenhouse gases (GHGs), odor-causing volatile organic compounds, dust and sediment associated with animal production facilities and manure application sites. Expansive research is going on to determine best management practices (BMPs) for crop production on land receiving agricultural waste with regard to nutrients and pathogens transport, runoff water, crop management, and soil types particularly in unique �karst topography�. Research is being conducted to look at how the addition of biochar to soil affects the transport behavior of Escherichia coli (E. coli) through unsaturated soil. Research is also being conducted on how biochar addition to soils affects the transport behavior of important pathogens commonly found in animal manures � Salmonella and Listeria � through soils. A study was initiated to evaluate the survival of naturally occurring pathogens (Campylobacter jejuni, Listeria monocytogenes and Salmonella enterica) and indicator organisms (E. coli, enterococci) in applied poultry litter or dairy manure. The first year of the study was completed. Concentrations of pathogens and indicators were monitored over the course of the study. Researchers also completed development of a high- throughput Deoxyribonucleic acid (DNA) extraction method for analysis of target organisms in broth and soil samples. A back grounding cattle operation was used as a model site for characterization of soil nutrients, microbial pathogens, and veterinary pharmaceutical spatial distribution in a feedlot situation. Work in conjunction with Agricultural Research Service (ARS) scientists in Beltsville, MD, and in New Orleans, LA, continues. A new study was initiated to evaluate swine manure composting. The study is being conducted in collaboration with a farmer in Davies County, KY, and with a University of Kentucky (UK) Extension specialist. Nutrient, bacteria and chemical analyses have been conducted on two composting runs. One additional study will be conducted to determine the effect of using corn refuse as a bulking agent on the compost process. Experiments were finished determining sorption coefficients for polar organic compounds in organic matter poor soil and in the same soil amended with animal waste-derived biochar. Continuing research was performed on determining the effect of wastewater recirculation through an aerated semi-permeable membrane on wastewater malodors and greenhouse gases. An experiment has begun in utilizing biochar from fast pyrolysis of rice- hull poultry litter in biofiltration systems for ammonia removal. The experimental objective was to compare the abiotic and biotic effects of biochar in removing ammonia from air stream. Lab-scale columns were packed with poultry litter, combination of poultry litter and biochar, and biochar alone for filtration of synthetic ammonia. Significant Activities that Support Special Target Populations: Fact Sheet �Amending poultry litter to reduce ammonia producing bacteria� targeted to farmers, integrators and U.S. Poultry & Egg association members. Fact Sheet �Johne�s Disease: Troughs as a hidden source� targeted to dairy farmers and members of the International Association for Paratuberculosis. Accomplishments 01 Atmospheric concentrations measured at a dairy lagoon. The ambient ion monitor (AIM) method was used to detect gas and particle phase amines at dairy lagoon in California. Previously, detection of reactive amines in- situ to agricultural waste management was only performed by detection of gases or aerosol phase and not both simultaneously. ARS scientists from Animal Waste Management Research Unit in Bowling Green, Kentucky, successfully performed the first in-situ amine detection at a dairy usin simultaneous gas- and aerosol- detection. Data comparisons are ongoing with collaborating scientist from ARS-Ames, Iowa, Cal State University- Fresno, and Claremont McKenna College. The study showed that some amines (e.g. trimethylamine) partition into the aerosol phase near source but others (e.g. butylamine) do not and are only observed in the gas-phase. This information benefits both the scientific community (e.g. atmospheri modelers) and potentially regulatory agencies to help determine which amines are potentially important to consider in atmospheric chemistry rather than focusing on all compounds. 02 Laboratory smog chamber studies of amines. An ambient ion monitor (AIM) and aerodynamic particle sizer (APS) were used to study the atmospheric reactions and aerosol forming potential of amines. Previous studies have shown that amines are capable of producing aerosol particles but it is unclear whether this is due to acid-base chemistry yielding salt formati in the atmosphere or whether it is due to photochemical reactions typica with urban sources of air pollution. ARS scientists from Animal Waste Management Research Unit in Bowling Green, Kentucky, took the AIM and AP instrumentation to Riverside, California, to perform laboratory studies atmospheric chemistry in the University of California Riverside�s (UCR) state-of-the-art environmental smog chamber. This project was collaborative with researchers from UCR, Utah State University, and Claremont McKenna College. The data show that both the acid-base and photochemical pathways for amines to form aerosol are important under normal atmospheric conditions. The data also indicate that there are som pathway differences that are influenced by relative humidity in the atmosphere. This project benefits the scientific community in helping to elucidate critical atmospheric chemistry questions. 03 Using phosphorus models to evaluate the accuracy of phosphorus indices. The U.S. Department of Agriculture�s Natural Resource Conservation Servi (USDA-NRCS) recently revised its 590 Nutrient Management Conservation Standard. As part of this revision, USDA-NRCS is requiring that states test the accuracy of their phosphorus index, a tool which is used to assess the vulnerability of phosphorous loss from agricultural fields, using either measured phosphorus loss data or data generated from a process-based phosphorus loss model. With this in mind, an ARS scientist at the Animal Waste Management Research Unit in Bowling Green, KY, used phosphorus loss data generated from a phosphorus transport model to evaluate the KY phosphorus index. Moreover, the general formulation of t KY phosphorus index was evaluated against current research on the processes controlling phosphorus transport in the environment. Results suggest that in some areas the KY phosphorus index does a good job in assigning phosphorus loss risk, however, this analysis also showed some important deficiencies in the index, primarily the neglect of important factors known to affect phosphorus loss (e.g., soil erosion and phosphor application rates) and how the different factors in the index are weight This research has resulted in the ARS scientist giving invited talks at universities, watershed advocacy group meetings, and local scientific meetings in KY. The ARS scientist has also been asked by KY USDA-NRCS an the University of KY to lead an effort to revise the KY phosphorus index 04 Use of biochar to increase Escherichia coli retention in soils. The incorporation of biochar into soils has been proposed as a means to sequester carbon from the atmosphere. Recent studies have shown added environmental benefits to biochar amendments to soil such as increased soil retention of nutrients, heavy metals, and pesticides, yet no studie have looked at the role biochar plays on microbial transport in soils. A scientists at the Animal Waste Management Research Unit in Bowling Green KY, have conducted studies to evaluate whether biochar amendments can reduce the transport of E. coli through soils. Results from this researc show that the addition of biochar to soil can indeed affect the retentio and transport of E. coli � a commonly used indicator of fecal contamination in groundwater. Results indicate that the amount of E. col retention in biochar-amended soils depends on biochar application rate, temperature that the biochar was produced, and surface characteristics o the bacteria. If biochar is to be widely used as a soil amendment, it is important that its impact on microbial transport through the soil be understood, especially for fields where animal manure is applied. This research is the first to investigate this important environmental topic and results from this research provide important information on the factors controlling bacterial transport through biochar-amended soils. Results suggest that biochar has the potential of being used as a management practice for protecting shallow groundwater supplies from contamination by pathogenic microorganisms. 05 Impact of growth conditions on transport behavior of Escherichia coli. coli is the most commonly identified fecal indicator bacteria originatin from agricultural and urban runoff, and is used as an indicator of water quality. Hence, understanding the fate and transport of E. coli originating from agricultural waste is essential for assessing water quality and protecting drinking, recreational, and agricultural water sources from contamination. Most published research on E. coli fate and transport has utilized highly idealized conditions; in particular, nutrient-rich growth media is often used for the organisms to grow in. Such nutrient-rich conditions, however, are not representative of animal manures. Therefore, to understand the fate and transport of E. coli originating from agricultural waste it is essential that the organisms a grown in more representative media. To address this gap in the scientifi literature, ARS scientists at the Animal Waste Management Research Unit Bowling Green, KY, in collaboration with the University of California at Riverside conducted a study to compare the differences in the fate and transport of E. coli cells grown in dairy manure extract solution and a more traditional laboratory growth media. Following growth in one of the two growth media, the surface properties of eleven E. coli isolates were extensively characterized and the transport of each isolate through 10-c columns packed with sand was measured. In general, cells grown in manure extract had different cell-surface properties and transport behavior tha cells grown in standard nutrient-rich media. This study shows that when studying E. coli transport, consideration must be given to utilizing growth conditions that better mimic the conditions that bacteria are exposed to in the environment. Utilizing more appropriate growth solutio will likely lead to a better understanding of the behavior of manure- derived bacteria in aquatic and terrestrial environments. 06 Atmospheric emissions of greenhouse gases (GHG) from different nitrogen fertilizers. Increasing demand for food and agricultural products directly relate to increased greenhouse gas (GHG) emissions, particularl the three primary gases associated with agriculture [Nitrous Oxide (N2O) Methane (CH4), and Carbon Dioxide (CO2)]. Commercial Nitrogen fertilizer and organic N sources such as animal manure stimulate N losses mainly through biochemical processes. ARS scientists at USDA-ARS in Bowling Gre Kentucky, have investigated the effects of N2O, CH4, and CO2 emissions from application of several inorganic N fertilizers, commercially available enhanced-efficiency N fertilizer, and poultry litter under no- till corn production. No significant differences were observed in N2O emissions among the enhanced-efficiency N fertilizers and other N fertilizer sources. The CH4 and CO2 emissions were impacted by the environmental factors more than the N source. Results demonstrated that fertilizer source and climate conditions need consideration when selecti N fertilizer that reduces greenhouse gas emissions. 07 Amending poultry litter to reduce ammonia producing bacteria. As fertilizer costs increase, poultry litter has become an increasingly valuable commodity. Reducing ammonia volatilization from poultry litter therefore important to not only reduce ventilation costs and improve bir performance but also to retain the fertilizer value of the litter. Ammon is produced when bacteria in the poultry litter breakdown poultry urine e., urea and uric acid) which makes up 80% of the nitrogen in poultry litter. ARS scientists in the USDA-ARS Animal Waste Management Research Unit Lab at Bowling Green, KY, and in the Richard B. Russell Research Center (RBRRC) in Athens, GA, found that poultry litter amendments reduc ammonia emissions by both chemical and microbiological means. These ARS researchers showed that acidifier amendments and a novel litter adsorber amendment, chitosan, reduced ammonia producing bacteria to the greatest extent. Chitosan is a derivative of chitin one of the most abundant sources of carbon on earth. This low cost, readily available, poly- cationic adsorbent used alone or in conjunction with an acidifier may delay onset of ammonia production thereby reducing organic nitrogen loss from litters. This research provides valuable new information for producers, integrators and researchers interested in reducing ammonia volatilization from poultry litter. 08 Inorganic fertilizers after broiler litter amendment reduce surplus nutrients in orchardgrass soils. Broiler litter is a good source of pla nutrients and as a soil amendment. It contains substantial amounts of nitrogen (N), phosphorus (P), potassium (K), and other nutrients, such a calcium, magnesium, sulfur, manganese, and zinc (Zn). Poultry producers usually dispose of broiler litter at high rates to forage crops, which allow excessive accumulation of soil nutrients. The ARS and Western Kentucky University scientists collaborated in a remediation study to examine if inorganic fertilizer application over the residual fertility broiler litter would reduce surplus soil nutrients by orchardgrass from nutrient enriched soil and by removing the resulting biomass from the study area. Three remediation treatments; inorganic fertilizer on antecedent broiler litter at the N rate, P rate, P rate with inorganic N and inorganic fertilizer were tested. Repeated broiler litter applicatio at N rate led to elevate soil P, copper (Cu), and Zn contents >200% from the initial levels. Implementing an alternative inorganic fertilizer application cycle in such soils can reduce P, Cu, and Zn levels respectively by 32 mg P kg-1 yr-1, 1.9 mg Cu kg-1 yr-1, and 2.4 mg Zn kg yr-1. Remediation by inorganic fertilizer application may take at least yrs to reduce P, Cu, and Zn levels back to the normal range. In addition inorganic fertilizer application over broiler litter at N rate can offer similar forage production as inorganic fertilizer and N rate broiler litter amendment. An alternative cycle of inorganic fertilizer applicati after broiler litter amendment can be recommended as a best management practice to remediate surplus soil nutrients in highly broiler litter impacted soils while assuring high forage production benefits. 09 High genotypic and phenotypic diversity of Escherichia (E.) coli in livestock manures and water sources. Many federal, state and local regulatory agencies use E. coli as an indicator for the presence of feca pathogens, whether bacteria, viruses, or protozoa. Understanding the ecology of this important organism is needed so more accurate protocols for monitoring may be developed. ARS scientists in the Animal Waste Management Research Unit (AWMRU) Lab at Bowling Green, KY, determined th genetic profiles of 1,346 E. coli isolates and transport properties of representatives from poultry, swine and cattle manure, and from a pollut creek in rural Kentucky. Working in collaboration with Kentucky Division of Water and Western Kentucky University, ARS researchers found that the was a high degree of diversity in the environmental transport potential and in genetic factors present in E. coli strains both within an environment (i.e. dairy manure) and between different environments (i.e. dairy manure and swine manure). This diversity should be taken into consideration by any researcher or regulatory agency when using only a f E. coli isolates for purposes of modeling, source tracking and risk assessment. 10 Soil nutrients, veterinary pharmaceuticals, and bacterial populations on cattle feedlot are concentrated in feeding and dirt areas. Beef cattle backgrounding operations that grow out weaned calves for feedlot finishi can become sources of environmental contaminants. Better understanding o these contaminants and their distribution will aid in development of effective contaminant management guidelines for sustainable livestock production. ARS scientists in the Animal Waste Management Research Unit (AWMRU) Lab at Bowling Green, KY, and the Environmental Management and B Product Utilization (EMBU) Lab in Beltsville, MD, in collaboration with scientists from Western Kentucky University investigated the distributio of soil nutrients, bacteria, and veterinary pharmaceutical compounds across a small cattle operation in western KY. Researchers showed that a contaminants were highly concentrated in the feeder area and were low in grass strips leading to a retention basin and sink hole. Grazing pasture within small feedlots can offer effective hydrologic isolation for contaminants. Therefore, contaminant management plans for small feedlots should focus on feeder areas where nutrients, bacteria and pharmaceutica are most concentrated. 11 Emission and plant uptake of trace elements and mercury (Hg) from application of flue gas desulfurization (FGD) materials as soil amendmen According to American Coal Ash Association, about 140 million metric to of coal combustion by-products are produced annually, including FGD gyps and waste byproducts from dry and wet scrubbers in the United States. Wh applied to soil as amendment, FGD gypsum is able to improve the yield of crops by providing plant nutrients, improve soil physical properties, increase water infiltration and storage, and reducing nutrient and sediment losses. ARS scientist in Bowling Green, KY, in collaboration wi scientists from Western Kentucky University conducted field study to investigate the distribution of mercury (Hg) and other potentially hazardous elements emission to ambient air and uptake by surface vegetation following application of different FGD materials to soil. We found Hg released into the air and uptake in grass from all FGD material treated soils was greater than the untreated soil. No Hg was detected in the leachate collected during the only 1-inch rainfall event that occurr throughout the 4-week testing period. In addition, this study demonstrat that considering only the amounts of trace elements uptake in surface vegetation may under estimate the overall release of the trace elements from FGD material-amended soils. It also shows, under the same soil conditions, the mobility of trace elements varies when FGD materials produced from different processes. 12 Heat fluxes and emission of malodorants from animal wastewater lagoons. Malodors from waste treatment lagoons are often a source of complaints. The concentrations of phenol-cresol and phenol-ethylphenol, two malodorants typical of animal manure were measured above a swine wastewater treatment lagoon. At the same time, measurement of air and water temperature, humidity, wind speed and solar radiation were also performed so that the environmental factors controlling release of malodors from waste treatment lagoons could be determined. The factors that best explained emission of these malodorants were evaporation from the lagoon surface and that portion of solar radiation available at the lagoon surface after subtracting that portion of the radiation used to heat the lagoon water. Emissions were found to be higher in the cool season than the warm season due to losses of malodorants that occurred early in the spring as the lagoon warmed. Results of this work are bein used to determine appropriate models to estimate malodorant emissions fr lagoons and devise techniques for the abatement of nuisance emissions. 13 Modeling emission of malodorants from waste treatment lagoons. Odors generated by large scale animal rearing operations are the foremost caus of complaints by people living in the vicinity. The wastes generated are largely stored and treated in facultative lagoons prior to land application. Using equations developed in a previous study, scientists with the Agricultural Research Service in Bowling Green, KY, and Western Kentucky University modeled emission of phenol-cresol, a typical manure malodorant, above a wastewater lagoon. The model, which relied on the variables of lagoon evaporation and radiation, predicted highest emissio of malodors during the months of March and April with steeply declining emissions thereafter. In this, the model agreed well with experimental data. During the study, there were no pronounced differences in microbia populations as determined by molecular quantification of bacterial genes Results of this study suggest evaporative losses are largely responsible for determining malodorous emissions from wastewater lagoons. 14 Improvement in biogas quality by wastewater recirculation through a silicone membrane. Biogas is a mixture of methane, carbon dioxide and trace gases such as hydrogen sulfide produced by the anaerobic breakdown of organic matter. The methane in biogas represents a severely underutilized resource from animal manures. The principal reasons that biogas is not harvested from animal wastes has to do with its corrosive nature and the susceptibility of anaerobic digests to operational upsets due to poor acid-base buffering. Researchers with the Agricultural Research Service in Bowling Green, KY, recirculated wastewater through a silicone hose located in an external aeration tank to improve the qualit of biogas produced by bacteria digesting swine manure. Carbon dioxide i biogas from treated swine wastewater was reduced by almost 80% compared control wastewater digestions. This result was due to a decrease in acidity of the treated wastewater. This also resulted in increased captu of carbon dioxide in the form of bicarbonate anion in the treated wastewater, increasing wastewater buffering and rendering treated wastewater less susceptible to operational upsets due to poor buffering. This result demonstrates a simple means for improving biogas quality and increasing operational efficiency of anaerobic digesters. 15 Utilization of poultry litter for pesticide bioremediation. Researchers from ARS unit in Bowling Green, KY, along with scientists from universit in Mexico conducted experiments to elucidate the stimulatory effect of poultry litter on the fungal growth. The goal of this study was to determine whether poultry litter could be utilized to support fungal and microbial growth in a mixture of pesticides. The results showed that poultry litter was able to enhance fungal growth better than vitamins in the presence of pesticide mixture. Therefore, poultry litter can serve growth substrate for fungal growth and provide additional capable microflora for pesticide bioremediation. This can reduce cost quite substantially when replacing the required vitamins for growth with poult litter at contaminated sites or treatment systems such as barrier walls. Based on these data, poultry litter can be used as nutrient source for bioremediation of pesticides in agricultural settings. As a result of th experimentation, application of this poultry amendment has been utilized on test plots and local crop lands with pesticide problems in Mexico. 16 Elucidating of the effect of different application methods on ammonia an Green House Gas (GHG) gas emissions. Researchers from ARS unit in Bowli Green, KY, along with scientists from the University of Kentucky conduct experiment to monitor the initial and subsequent ammonia and greenhouse gas emissions from three different liquid swine manure application metho at a farm in Kentucky which had been cropped in a no-till corn/soybean rotation, using flux chambers and a gas analyzer. Results showed that Surface spray method has the highest total greenhouse potency factor bas on non-linear method of calculating flux follow by Aerway and Row injection methods during the first four hours after application. The Surface spray method emitted about four times more than the Aerway injection and about 31 times more than the row injection method in greenhouse potency factor. The Aerway injection method emitted greenhou potency factor of about seven times more than the row injection. Based these data, the types of swine effluent application can have a significa initial impact on the ammonia and greenhouse gas emissions. This is very critical in reducing ammonia and GHG emissions via different types of la application methods. The results from this experiment will serve as the guideline for delineating potential factors that may affect the emission from these various land application methods of liquid livestock wastes.

Impacts
(N/A)

Publications

Sistani, K.R., Jn-Baptiste, M., Lovanh, N.C., Cook, K.L. 2011. Atmospheric emissions of N2O, CH4, and CO2 from different nitrogen fertilizers. Journal of Environmental Quality. 40:1797�1805.
Bolster, C.H., Abit Jr, S.M. 2012. Biochar pyrolyzed at two temperatures impacts E. coli transport through a sandy soil. Journal of Environmental Quality. 41(1):124-133.
Marcus, I.M., Bolster, C.H., Cook, K.L., Opot, S.R., Walker, S.L. 2012. Impact of growth conditions on transport behavior of E. coli. Journal of Environmental Monitoring. 14:984-991.
Tewolde, H., Sistani, K.R., Rowe, D.E. 2005. Broiler litter as a micronutrient source for cotton: concentrations in plant parts. Journal of Environmental Quality. 34:1697-1706.
Bjerklie, D.M., Dingman, L., Bolster, C.H. 2005. River discharge and general flow resistance in the manning and chezy equation revisited. Water Resources Research. 41, W11502.
Cook, K.L., Garland, J.L., Layton, A.C., Dionisi, H.M., Levine, L.F., Sayler, G.S. 2007. Effect of microbial species richness on community stability and community function in a model plant-based wastewater processing system. Microbial Ecology. 52:725-737.
Loughrin, J.H., Szogi, A.A., Vanotti, M.B. 2006. Reduction of malodorous compounds from liquid swine manure by a multi-staged treatment system. Applied Engineering in Agriculture. 22(6):867-873
Tewolde, H., Shankle, M.W., Sistani, K.R., Adeli, A., Rowe, D.E. 2008. No- till and conventional-till cotton response to broiler litter fertilization in an upland soil: lint yield. Agronomy Journal. 100:502-509.
Quintanar, A., Mahmood, R., Loughrin, J.H., Lovanh, N.C. 2008. A coupled mm5-noah land surface model-based assessment of sensitivity of planetary boundary variables to anomalous soil moisture conditions. Physical Geography. 29(1):54-78
Quintanar, A., Mahmood, R., Motley, M., Yan, J., Loughrin, J.H., Lovanh, N. C. 2009. Simulation of boundary layer trajectory dispersion sensitivity to soil moisture conditions: MM5 and noah-based investigation. Atmospheric Environment. 43:3774-3785.
Loughrin, J.H., Vanotti, M.B., Szogi, A.A., Lovanh, N.C. 2009. Evaluation of second-generation multistage wastewater treatment system for the removal of malodorous compounds from liquid swine waste. Journal of Environmental Quality. 38:1739-1748.
Cheng, C., Chang, Y., Sistani, K.R., Wang, Y., Lu, W., Lin, C., Dong, J., Hu, C., Pan, W. 2012. Mercury Emission and Plant Uptake of Trace Elements during Early Stage of Soil Amendment Using Flue Gas Desulfurization Materials. Journal of Air and Waste Management Association. 62(2):139-150.
Tewolde, H., Shankle, M., Adeli, A., Sistani, K.R., Rowe, D.E. 2009. Macronutrient concentration in plant parts of cotton fertilized with broiler Litter in a marginal upland Soil. Soil & Tillage Research. 105(1) :1-11.
Sistani, K.R., Adeli, A., Tewolde, H. 2010. Apparent use efficiency of nitrogen and phosphorus from broiler litter applied to bermudagrass. Communications in Soil Science and Plant Analysis. 41(15):1873-1884.
Doerner, K., Cook, K.L., Mason, B. 2009. 3-Methylindole production is regulated in Clostridium scatologenes ATCC 25775. Letters in Applied Microbiology. 48:125-132.

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

Outputs
Progress Report Objectives (from AD-416) The overall goal of the research project which is formulated as a real partnership between ARS and Western Kentucky University (WKU) is to conduct cost effective and problem solving research associated with animal waste management. The research will evaluate management practices and treatment strategies that protect water quality, reduce atmospheric emissions, and control pathogens at the animal production facilities, manure storage areas, and field application sites, particularly for the karst topography. This Project Plan is a unique situation in the sense that non-ARS scientists from WKU are included on an in-house project to conduct research under the NP 214. The objectives and related specific sub-objectives for the next 5 years are organized according to the Components (Nutrient, Emission, Pathogen, and Byproduct) of the NP 214, which mostly apply to this project as follows: 1) develop improved best management practices, application technologies, and decision support systems for poultry and livestock manure used in crop production; 2) develop methods to identify and quantify emissions, from poultry, dairy and swine rearing operations and manure applied lands; 3) reduce ammonia, odors, microorganisms and particulate emissions from dairy, swine and poultry operations through the use of treatment systems (e.g. biofilters and scrubbers) and innovative management practices; 4) perform runoff and leaching experiments on a variety of soils amended with dairy, swine, or poultry manures infected with Campylobacter jejuni (C. jejuni), Salmonella sp. or Mycobacterium avium subsp. paratuberculosis (MAP) and compare observed transport with that observed for common indicator organisms such as E. coli, enterococci, and Bacteriodes; and 5) use molecular-based methodologies to quantify the occurrence of pathogens and evaluate new methods to inhibit their survival and transport in soil, water, and waste treatment systems. Approach (from AD-416) This in-house project was conceived as a cooperative/partnership and comprehensive research program between USDA-ARS Animal Waste Management Research Unit (AWMRU) and Western Kentucky University (WKU). The project is designed to utilize the scientific expertise and facilities of both institutions to conduct problem-solving research related to animal waste management in Kentucky and the southeastern U.S. The research effort will be multi-disciplinary and multifaceted in support of decision making and systems development. Research focuses will be on all four components (Nutrient, Atmospheric Emission, Pathogens, and By-products) of the National Program 214. In lieu of repeatedly stating the equipment used for analysis, please note that the state-of-the-art laboratories and equipments exist at both AWMRU and WKU, which can be accessed by the scientists including land at the WKU research station. Main instruments include: ICP, GC-MS, 2 GCs, Latchet, 2 C/N Analyzers, IC, HPLC, Real-time PCR, etc. A 2 year study was initiated to evaluate the survival of naturally occurring pathogens (Campylobacter jejuni, Listeria monocytogenes and Salmonella enterica) and indicator organisms (Escherichia coli (E. coli), enterococci) in applied poultry litter or dairy manure. Preliminary studies were conducted to determine the microbial population in the manures and to evaluate culture and molecular methods to follow the organisms. Tall fescue plots with applied manures were established and samples taken for chemical and microbial analysis. A back grounding cattle operation located on Western Kentucky University was used as a model site for characterization of soil nutrients, microbial pathogens, and veterinary pharmaceutical spatial distribution in a feedlot situation. A Global Positioning System (GPS) ground survey was completed and five sets of samples taken for analysis from soils and run-off. Research is being conducted to look at whether the addition of biochar to soil affects the transport behavior of Escherichia (E.) coli through water-saturated and unsaturated soil. Research is also being conducted on the affect of biochar amendment on the sorption of organic compounds to soils. We also performed sorption isotherm experiments for polar organic compounds using swine, cow, and turkey waste biochar prepared at low (350� centigrade) and high (700�). We found that animal waste biochar can enhance binding of the polar organic compounds to soil. Research is also being conducted on ways for improving phosphorus indices. As part of this research, modeling work has been conducted to identify limitations with the Kentucky phosphorus index. We built larger scale versions of novel anaerobic digesters in which waste is recirculated through a silicone membrane located in a supplemental aeration tank. We determined that the system improves biogas quality by increasing bicarbonate buffering and lowering carbon dioxide as well as hydrogen sulfide concentrations. We are utilizing biochar from fast pyrolysis of rice-hull poultry litter in biofiltration systems for ammonia removal. The experimental objective was to compare the abiotic and biotic effects of biochar in removing ammonia from air stream. In another experiment, lab-scale trickling filters seeded with swine micro flora were tested to see their efficiency and efficacy in removing skatole (main odor causing compound from swine waste). The results obtained from this experiment will be used to scale a pilot-scale size bioscrubber. Initial field test of Ambient Ion Monitor (AIM) instrument was successfully conducted; amines were detected in both the particle and gas phase, but during a relatively clean atmospheric event. Continued method development is necessary before direct emission studies can be conducted because air concentrations are expected to be much higher in near-source emission studies and may overwhelm detector. Significant Activities that Support Special Target Populations Scientist served on a special committee to make recommendations to Natural Resources Conservation Service (NRCS) for updating their 590 standards. This led to two white papers that NRCS is using as they make modifications to their 590 standard. Minority PhD Candidate from Alabama A&M University working on-site at Agricultural Research Service (ARS)-Animal Waste Management Research Unit (AWMRU).

Impacts
(N/A)

Publications

Netthisinghe, A., Gilfillen, B., Willian, T., Rowland, N., Sistani, K.R. 2011. Inorganic fertilizers after broiler litter amendment reduce surplus nutrients in orchardgrass soils. Agronomy Journal. 103:1-8.
Cook, K.L., Rothrock Jr, M.J., Eiteman, M.A., Lovanh, N.C., Sistani, K.R. 2011. Evaluation of amendments to manage nitrogen loss and microbiological quality in poultry litter. Journal of Environmental Management. 92:1760- 1766.
Tellinghuisen, J., Bolster, C.H. 2011. Using R^2 to compare least-squares fit models: When it must fail. Chemometrics and Intelligent Laboratory Systems. 105:220-222.