2008 Annual Report 1a.Objectives (from AD-416) Increase the current effort to develop and evaluate management practices and treatment technologies that reduce air emissions of ammonia and odor causing compounds from animal production operations, manure storage areas, and field application sites. The overall goal of the research project formulated in 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 unique karst topography. This Project is a unique situation in the sense that non-ARS scientists from a university are included in a research project to conduct research under the same National Program. Hence, to achieve the ultimate goal of this project, the integration and coordination of scientific expertise of the scientists from ARS and WKU are required within and across all objectives. The objectives and related specific sub-objectives are organized according to the three major components (Nutrient, Emission, and Pathogen) of the National Program 206, which mostly apply to this project. The specific objectives for the next 5 years are: Nutrient Component Objective 1: Develop management practices and decision tools for long-term use of animal manure as an alternative source of fertilizer for forages and row crops with regard to the following factors: Impacts on crop yield, nutrient loading, availability and uptake, application rate and timing, tillage, methods of application, soil quality, and soil carbon sequestration and greenhouse gas emissions. Objective 2: Determine if nutrient loading from agricultural watersheds in karst terrain is a function of physical watershed characteristics. Emission Component Objective 3: Reduce odiferous emissions by developing innovative molecular-based methods to identify and quantify microorganisms and biological activities responsible for production of odorous compounds in livestock wastes. Objective 4: Develop new analytical approaches to quantify gases (e.g. methane, H2S), volatile odor compounds (e.g. p-cresol, skatole, and other VOCs) and evaluate treatment technologies for odor abatement at animal production facilities and manure-applied fields. Pathogen Component Objective 5: Employ molecular-based methods to improve detection, quantification, and evaluation of transport, and survival of pathogens including Salmonella and E. coli O157:H7 from animal manure. Also, compare survival of these pathogens with indicator organisms through a series of laboratory and watershed studies. 1b.Approach (from AD-416) This research 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 research is designed to utilize the scientific expertise and facilities of both institutions to conduct problem-solving research related to the animal waste management in Kentucky and the Southeastern US. The research effort will be multi-disciplinary and multifaceted in support of decision making and systems development. Research focuses will be on all three components (Nutrient, Atmospheric Emission, and Pathogens) of the National Program 206. State-of-the-art laboratories and equipments exist at both AWMRU and WKU, which can be accessed by the scientists. Main instruments include: ICP, GC-MS, Lachat, C/N Analyzer, Real time PCR, etc. 3.Progress Report Research activities continued under Problem Area 1 (Methods Assessment and Development) of the NP 206 as follow: 1. Identification of Odor Producing Microorganisms in Livestock Wastes: We measured the temporal changes in total bacterial and fungal populations and urease-producing microorganisms in untreated litter or litter treated with 10% alum. In these incubation studies, nitrogen mineralization occurred concomitant with increases in bacterial urease producers in normal litter and with increases in fungal urease producers in alum-treated poultry litter. The decrease in pH produced by alum treatment is believed to inhibit bacterial populations and favor growth of fungi that may be responsible for the mineralization of organic nitrogen in alum-treated litters. 2. Evaluate Pathogen Survival and Transport. Campylobacter sp and Salmonella sp. are the leading causes of gastrointestinal illness worldwide. Molecular methods were used to determine the level of C. jejuni in litter from three states; to evaluate the effect of alum amendment on survival of the pathogen in naturally infected litter; and to assess the transport of C. jejuni through repacked soil columns. Data from these studies suggest that C. jejuni is present in poultry litter from many sources and can be transported from the litter during rainfall events. However, treatment with litter additives such as alum and allowing contaminated litter to dry following application and prior to rainfall should minimize associated health risks. 3. Trough’s as a Hidden Source of Johne’s Disease. Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease, a chronic enteric infection that affects ruminants. The goal of this study was to evaluate the ability of MAP to become established and persist in biofilms on coupons composed of common livestock watering trough materials. Results show that trough material composition influenced the survival of MAP with the lowest survival exhibited on stainless steel, followed by plastic, galvanized steel and concrete. 4. Karst transport. Research is ongoing looking at the transport behavior of agricultural contaminants in karst environments. This work includes injecting nutrient salts into underground karsts streams and monitoring their movement downstream. The work also includes the tracing of manure particles through shallow karst soils. 5. Evaluation of Second Generation Environmentally Superior Technology for The Reduction of Malodors from Liquid Swine Waste: Malodors and water quality of wastewater at successive stages of a modular waste treatment system installed on a 5,145 head swine finishing farm were monitored over 15 months. Large improvements in both water quality and odor were consistently achieved. Up to 250 kg of aromatic malodors and up to 30 metric tons of VFA were removed every month by the second generation EST. 6. Bench and Pilot Scale Testing of Treatment Systems: Testing of biofilter is underway in examining different compost-size packing materials for ammonia removal assessment. Pilot scale scrubbers are being constructed to examine the chemical and biological remediation of ammonia and malodors. 4.Accomplishments 1. Managing Broiler Litter Application Rate and Grazing to Decrease Watershed Runoff Losses: It has been reported that over 11.4 million tons of broiler litter was generated in 1996, of which over 90% was land applied. With regard to permanent pasture systems, inability to incorporate waste materials leads to increased nutrient concentration, such as P, Cu, and Zn near the soil surface. We studied the impact of pasture management and broiler litter application rate on runoff nutrient concentration. The inorganic N content of the runoff exiting grazed or grazed/hayed combination paddocks with litter applied on N basis was greater than the same treatments that received litter on P basis. All the treatments that received broiler litter on N basis had greater total P and total dissolved P in the runoff water. The mean total P loss per runoff event ranged from 7 to 45 g ha-1, and the grazed treatment with litter applied on N basis had the most. Total dissolved P was the dominant P fraction in the runoff ranging from 85% to 93% of the total P. Our results indicate that litter should be applied on N basis if the soil P is low and the pasture is used for hay production. In contrast, litter should be applied on P basis if pasture is being used for grazing. The accomplishment addresses Problem Area 4 -- Farming Systems and Practices for Efficient and Balanced Manure Nutrient Management -- of the Nutrient Management Component under NP 206. 2. Broiler Chicken Litter Application Timing Effect on Coastal Bermudagrass in Southeastern U.S: In many states swine effluent application timing is regulated and limited to spring and summer months. In contrast, poultry litter application timing is not regulated; hence it is being surfaced land applied year-round in many states. It is expected that soon, litter application during the winter months or rainy months will be prohibited. Therefore, producers have to store litter for application during spring and summer. In this study, we attempted to identify an optimum date or dates for litter application that maximizes nutrient availability and uptake to crops and minimizes nutrient buildup in soil or losses through runoff. All the litter application dates did not influenced coastal bermudagrass dry matter yield and nutrient uptake except for nitrogen. The residual buildup of carbon, which is a beneficial impact, and the phosphorus, cupper, and zinc with potential negative impact were observed with all litter application timings. We were expecting that late summer and early fall litter application to bermudagrass would be less efficient than early spring and early summer application dates, but high soil fertility level at the study site and climatic factors may have impacted the costal bermudagrass growth, which confounded the real response to litter application timing. The accomplishment addresses Problem Area 4 -- Farming Systems and Practices for Efficient and Balanced Manure Nutrient Management -- of the Nutrient Management Component under NP 206. 3. Identification of Odor Producing Microorganisms in Swine Wastes. Hydrogen sulfide, produced by sulfate reducing bacteria (SRB), is one of the most potent malodors emitted from anaerobic swine waste storage systems. However, little is known about the prevalence and diversity of SRB in those systems. In this study, the SRB population in swine manure storage systems was characterized and four new quantitative, real-time polymerase chain reaction (PCR) assays were developed to target SRB in swine slurry. To date, this is the only study to evaluate the phylogeny and concentration of SRB in any livestock waste storage system. This research addresses Problem Area 1 (Understanding the Biological, Chemical, and Physical Mechanisms Affecting Emissions) of Atmospheric Emissions Component of the National Program 206 - Manure and Byproduct Utilization. 4. Laboratory and Field Evaluation of Broiler Litter Nitrogen Mineralization: Broiler litter, a mixture of manure and bedding materials, is used widely as fertilizer on pasture and row crop lands in poultry producing regions. Most of the land applied litter is broadcasted on the pasture lands or incorporated for some row crops. Accurate estimation of litter nutrients is required for efficient utilization of this resource in agriculture. It is difficult to predict the availability of litter nitrogen (N) to plants since both N transformation (conversion of organic N to inorganic N) and losses of N influence availability. Limited evidence suggests that soil properties may affect the amount of net N availability from broiler litter. Two studies were conducted: first, a laboratory incubation study to quantify poultry litter N availability at two temperatures (18° and 25° C), two soil moisture regimes, constant and fluctuating, and three soil types, Brooksville silty clay loam and Ruston sandy loam from Mississippi, and Catlin silt loam from Illinois; second, a field study using only Brooksville and Ruston soil to validate the laboratory results, using similar soils in microplot cylinders and anion exchange resin to capture available N. The impact of temperature was significant, while soil moisture regimes had no significant impact on litter-derived inorganic N. Field results followed the same trend as the laboratory study but with much lower rate. The accomplishment addresses Problem Area 4 -- Farming Systems and Practices for Efficient and Balanced Manure Nutrient Management -- of the Nutrient Management Component under NP 206. 5. Understanding the role starvation status plays on the adhesion and transport properties of Escherichia coli in saturated porous media. Escherichia coli (E. coli) is commonly used as an indicator of fecal pollution. Therefore, it is important to have a good understanding of how this microorganism moves in the environment. Cell surface characteristics such as surface charge and hydrophobicity have been shown to play an important role controlling microbial movement in the environment. What is not well known is how these surface characteristics are affected by the starvation status of the cells. Research showed that surface properties and transport behavior of E. coli depended strongly on the starvation status of the cells. This study highlights the importance of consistency in bacterial preparation for transport and characterization studies and has considerable implications for the future evaluation and prediction of E. coli fate in subsurface environments. This research supports Focus Area 2a of the Pathogens Component of NP 206 – Inactivation Rates and Transport Characteristics of Pathogens in Animal Agriculture. 6. Equilibrium Sampling Used to Monitor Malodors in a Swine Waste Lagoon. The concentrations of malodorous compounds in an anaerobic swine waste lagoon were monitored during the late summer to late fall of 2006. While the lagoon was being pumped down, malodorous compounds fluctuated widely, then increased greatly as compared to the period before pumping, and continued to increase as fall progressed. Suspended solids, volatile suspended solids, and total organic carbon increased near the bottom of the lagoon during this same period. This research addresses Problem Area 1 (Understanding the Biological, Chemical, and Physical Mechanisms Affecting Emissions) of Atmospheric Emissions Component of the National Program 206 - Manure and Byproduct Utilization. 7. Effect of Stratification and Seasonal Variability on the Profile of an Anaerobic Swine Waste Treatment Lagoon. An understanding of the nature of wastes in an anaerobic swine lagoon is essential in the design and operation of alternative collection, treatment, and disposal facilities for environmental quality management such as odor control, nutrient and pathogen reduction. In this study, the characterization of an anaerobic swine waste treatment lagoon from a farrowing operation (~2000 sows) was carried out to examine the dynamics of the system due to stratification and seasonal variability. The sampling process was carried out from spring to fall season. The pH and temperature were monitored and recorded continuously from the epilimnion (top) and hypolimnion (bottom) layers of the lagoon. Results showed that nutrient (C, N, P, S) concentrations varied according to stratified lagoon layers and season. Trace minerals such as Al, Ca, Fe, K, Na, and Mg, on the other hand, appeared to be affected more by stratification than seasonal variability. The reason for the decrease in nutrient concentrations in summer time may be due to increase microbial activities which required more essential nutrients (i.e., C, N, P, S) rather than trace minerals for growth during active season. Microbial community structure appeared to be affected by the stratification and seasonal variability. The microbial community appeared to be more diverse in the spring time than late summer or early fall. Based on these data, it is important to consider the effect of stratification and seasonal variability of waste loading from traditional anaerobic swine lagoon when designing and operating an alternative anaerobic digester. This research addresses Problem Area 1 (Understanding the Biological, Chemical, and Physical Mechanisms Affecting Emissions) of Atmospheric Emissions Component of the National Program 206 - Manure and Byproduct Utilization. 6.Technology Transfer Number of New Commercial Licenses Executed 10 Number of Web Sites Managed 3 Review Publications Bolster, C.H., Hornberger, G.M. 2007. On the use of Linearized Langmuir Equations. Soil Science Society of America Journal. Vol.71 Issue 6 P 1796-1806 Sistani, K.R., Brink, G.E., Oldham, J.L. 2008. Managing Broiler Litter Application Rate and Grazing to Decrease Watershed Runoff Losses. Journal of Environmental Quality. 37:718-724 Sistani, K.R., Adeli, A., Tewolde, H., Brink, G.E. 2007 Broiler chicken litter application timing effect on coastal bermudagrass in southeastern u.s.. Nutrient Cycling in Agroecosystems. 81:49-57 Sistani, K.R., Adeli, A., McGowen, S.L., Tewolde, H., Brink, G.E. 2007. Laboratory and field evaluation of broiler litter nitrogen meralization. Bioresource Technology. 99:2603-2611 Sistani, K.R., Rasnake, M., Sikora, F. 2008. Poultry Litter and Tillage Influence on Corn Production and Soil Residual Nutrients 98:130-139 . Soil & Tillage Research. Warren, J.G., Penn, C.J., Mcgrath, J.M., Sistani, K.R. 2008. The Impact of Alum Additions on Organic P Transformations in Poultry Litter and Litter Amended Soil. Journal of Environmental Quality. 37:469-476 Griffin, T.S., Honeycutt, C.W., Albrecht, S.L., Sistani, K.R., Torbert Iii, H.A., Wienhold, B.J., Woodbury, B.L., Hubbard, R.K., Powell, J.M. 2008. Nationally coordinated evaluation of soil nitrogen mineralization rate using a standardized aerobic incubation protocol. Communications in Soil Science and Plant Analysis. 39:257-268. Cook, K.L., Whitehead, T.R., Spence, C., Cotta, M.A. 2008. Evaluation of the sulfate-reducing bacterial population associated with stored swine slurry. Anaerobe. 14:172-180. Haznedaruglu, B.Z., Bolster, C.H., Walker, S.L. 2008. The Role of Starvation on Escherichia coli Adhesion and Transport in Saturated Porous Media. Water Research. Vol 42 pp 1547-1554 Rothrock Jr, M.J., Cook, K.L., Lovanh, N.C., Warren, J.G., Sistani, K.R. Quantification of a Novel Group of Ammonia Producing Bacteria Found in Poultry Litter by Quantitative PCR. Poultry Science. 87:DOI:10.3382/ps2007-00350 Adeli, A., Bolster, C.H., Rowe, D.E., McLaughlin, M.R., Brink, G.E. 2008. Effects of long-term swine effluent application on selected soil properties. Soil Science. 173:223-235.