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United States Department of Agriculture

Agricultural Research Service


Location: Food Animal Environmental Systems Research Unit

2009 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
1) Poultry litter amendment studies evaluated the effectiveness of 3 acidifier amendments, 4 chemical treatments, and 1 biological treatment for reducing ammonia emissions. This work is completed and will be written up this year. 2) We evaluated the ability of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) the causative agent of Johne’s disease, to become established and persist on surfaces of common livestock watering trough materials (plastic, concrete, galvanized and stainless steel). This work is accepted for publication and will be included in a fact sheet entitled “Livestock watering troughs and Johne’s disease”. 3) Quantitative, real-time Polymerase Chain Reaction (PCR) assays are being designed to target methanogenic populations in soils and waste storage systems. These assays will be used to evaluate how manure storage and/or soil application effect methanogens and Green House Gas (GHG) production. 4) Research is being conducted to look at the role experimental conditions have on the diversity in transport behavior among E. coli isolates obtained from a swine lagoon. 5) A model was determined for the evaporation of water from the lagoon surface that correlated well with the volatilization of the malodorants from a waste lagoon surface. It was found that sunlight, by heating the upper layers of the lagoon surface, caused volatilization of both water vapor (latent heat) and malodors. It was also found that fluxes of malodorants declined dramatically in the spring due to declining source levels in the lagoon. 6) A sampler was tested to obtain wastewater samples for quality and gas analyses without atmospheric contamination or off-gassing of samplers. This sampler was used to determine source levels of greenhouse gases and malodorants for continuing flux studies. Samples will also be used for wastewater quality and microbial analyses. Research is being used to help determine the relative importance of malodorant losses due to volatilization and remediation to wastewater odor improvements during warm seasons. 7) Pilot-scale biofilters are being built to test their efficiency and efficacy in remediate potential air pollution from livestock operations. Testing of biofilter is underway in examining different compost-size packing materials for ammonia removal assessment. This technology could be a potential abatement strategy for treating polluted air streams from poultry operations. 8) Pilot scale scrubbers are also being constructed to examine the chemical and biological remediation of ammonia and malodors from livestock operations. In the near future, tests will be carried out on these scrubbers in removal of ammonia from synthetic and natural polluted air that mimic polluted air streams from poultry and swine operations. 9) Two experiments evaluating the environmental and agronomic benefits of subsurface application (subsurface banding) of poultry manure to permanent pasture were completed and results were published in the peer-reviewed scientific journals.

1. Optimization of Molecular Methods for Detection of Campylobacter jejuni in Water and Soil. Campylobacter jejuni is one of the most common causes of gastroenteritis in the world. Given the potential risks to human, animal, and environmental health, the development and optimization of methods to quantify this important pathogen in environmental samples is essential. ARS scientists at the Bowling Green, KY, AWMRU determined the limit of detection and the level of resolution for quantification of C. jejuni and compared this with that of the common indicator organism Escherichia coli. The use of selective plate count media for quantification of C. jejuni resulted in a 0.7–1.2 log under-estimation of cell concentrations, compared with qPCR in both water and soils, whereas E. coli concentrations were found to be similar with either technique. Only the qPCR assay accurately measured 2-fold changes in C. jejuni cell concentrations in water samples. These results suggest that currently used plating techniques are inadequate for measuring C. jejuni concentrations and these data further bring into question the adequacy of using E. coli as an indicator of this important pathogen.

2. New Method of Poultry Litter Application to Perennial Pasture, Subsurface Banding. More than 90% of the poultry manure generated annually is land applied by the conventional method of “surface-broadcast” application. Runoff from agricultural lands with surface-applied animal manure is a major source of non-point source pollution, inability to incorporate animal manure into permanent pasture, leads to increased nutrient concentration, such as P, Cu, and Zn near the soil surface. ARS scientists from Bowling Green, Kentucky, and Auburn, Alabama, locations studied the impact of a new litter application method, “sub-surface banding” on bermudagrass and tall fescue, two dominant forages in the southeastern region, in order to investigate forage yield, nutrient uptake, and nutrient losses through runoff water in comparison to the conventional surface-broadcast application of poultry litter. This research provided the first source of scientific information on the use of subsurface banding of poultry manure for permanent pasture to farmers and scientific community. Subsurface banding of poultry manure is not only a feasible practice, but it also offers many agronomic and environmental advantages compared to the traditional surface-broadcast method. Based on information gathered from extension agents in the region and interest shown by farmers during field demonstration, this practice will be adopted by many farmers, pending availability of the new implement.

3. Development of Second Generation Environmentally Superior Technology for the Remediation of Odor from Livestock Waste. Malodors from confined animal feeding operations are a source of complaints and may have adverse environmental impacts. ARS Scientists from Bowling Green, KY, and Florence, SC, studied reductions in malodor effected by a second generation wastewater treatment plant during three cycles of pig rearing. The wastewater treatment system consisted of three modules: solids separation, biological nitrogen removal, and phosphorus recovery/wastewater disinfection. While approximately over 90% of the wastewater solids were removed in the first stage of treatment, little reduction in malodorous compounds occurred, indicating that malodors largely remained with the liquid waste stream. The greatest improvements in wastewater quality occurred in the nitrogen treatment module: there was over 90% reduction in malodors as compared to the raw flushed manure. The system consistently achieved high performance standards, even during the first cycle of livestock production when system performance was being optimized. These findings showed that the combination of two simple processes into a practical treatment system can be very effective in reducing malodors from livestock wastewater.

4. Assessing the Diversity in Cell Properties and Transport Behavior Among 12 Different Environmental Escherichia coli Isolate. Escherichia coli is a commonly used indicator organism for detecting the presence of fecal-borne pathogenic microorganisms in water supplies. The importance of E. coli as an indicator organism has led to numerous studies looking at cell properties and transport behavior of this microorganism. In many of these studies, however, only a single strain of E. coli was used even though research has shown a significant amount of genetic variability exists among different strains of E. coli. If this genetic diversity results in differences in cell properties that affect transport, different strains of E. coli may exhibit different rates of transport in the environment. Research was conducted at the Animal Waste Management Unit (AWMRU), in collaboration with the University of California – Riverside, to assess the variability in surface characteristics and transport behavior of E. coli isolates obtained from six different sources. Results of this study showed a large diversity exists in cell properties and transport behavior for the different E. coli isolates. Because USEPA recommends the use of E. coli as an indicator of fecal contamination in freshwater systems, its fate and transport in the subsurface needs to be better understood. This study shows that generalizations about the transport behavior of E. coli based on results from a single strain of E. coli should be made with caution. In addition, this large variability suggests that the modeling of E. coli in the environment will require a distribution of bacterial attachment rates, even when modeling E. coli movement from a single fecal source.

5. Effect of Litter Acidification on Microorganisms that Produce Ammonia. Microbial mineralization of urea and uric acid in poultry litter results in the production of ammonia, which can lead to decreased poultry performance, malodorous emissions, and loss of poultry litter value as a fertilizer. Despite the fact that this is a microbial process, little is known about how the microbial populations, especially ammonia-producing (ureolytic) organisms, respond to litter amendments such as aluminum sulfate (Al2(SO4)3•14H2O; alum). ARS scientists at the Bowling Green, KY, AWMRU found that nitrogen mineralization in poultry litter occurred in conjunction with increases in bacterial urease producers in normal litter and with increases in fungal urease producers in alum-treated poultry litter. Alum amendment reduced ammonia emissions by both biological (inhibition of ureolytic microorganisms) and chemical means (conversion of NH3 to NH4-N). 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. This information should aid farmers and manufacturers of litter amendments by providing information about the organisms responsible for ammonia production and the affect of litter amendments on the microbial populations.

6. Transport Behavior of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) Through Soil and Aquifer Material. M. paratuberculosis is the causative agent of Johne’s disease, a chronic enteric infection causing diarrhea and wasting in cattle, sheep, and other ruminants which usually results in the death of the animal. Exposure of susceptible animals to M. paratuberculosis contaminated manure in the environment poses a significant risk for disease transmission as Johne’s disease is spread by ingestion of the organism, yet very little is known about the movement of this important pathogenic microorganism in the environment. Research was conducted at the Animal Waste Management Research Unit to better understand the movement of M. paratuberculosis in the soil and aquifer materials. Research found that Map has a strong negative charge and is highly hydrophobic and that the transport of Map through positively-charged Fe-coated sands was reduced compared to transport through negatively-charged clean quartz sand, though Map transport for all treatments was low compared to the transport behavior reported in the literature for other bacteria. The results suggest that the potential for groundwater contamination by M. paratuberculosis is low; however, the organism may remain bound to the soil near the surface where it can be ingested by grazing animals or be released during run off events. This was the first study looking at the surface characteristics and transport behavior of M. paratuberculosis through aquifer materials and, therefore, provides important information for understanding the movement of M. paratuberculosis in the environment.

7. Effect of Stratification and Seasonal Variability on the Profile of an Anaerobic Swine Waste Treatment Lagoon. Anaerobic lagoons are used for the storage and treatment of high strength organic wastewater from concentrated animal feeding operations (CAFOs). They 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. Although these anaerobic lagoon systems are able to reduce organic matters, nutrients, and other heavy metals, their efficiency is dependent on seasonal variability and other environmental conditions. Researchers from ARS unit in Bowling Green, KY, along with scientists from Western Kentucky University conducted experiments to elucidate the effect of stratification and seasonal variability on the profile of these anaerobic swine waste treatment lagoons. Results showed that nutrient (C, N, P, S) concentrations varied according to stratified lagoon layers and season. Trace minerals (Al, Ca, Fe, and Mg), on the other hand, appeared to be affected more by stratification than seasonal variability. Molecular analysis also showed that microbial community structure appeared to be affected by the stratification and seasonal variability. It is important to consider the effect of stratification and seasonal variability in managing these open lagoons when considering alternative treatment systems. The results from these experiments will serve as the guiding principles for delineating potential factors that may affect the emissions from these retaining/treatment receptacles.

8. Development of Improved Regression Methods for Fitting the Langmuir Model to Sorption Data. One of the most commonly used models for describing phosphorus sorption to soils is the Langmuir model. However, several potential problems exist when fitting the Langmuir model to sorption data using traditional methods. An ARS scientist at Animal Waste Management Research Unit examined commonly used regression methods for fitting the Langmuir model to sorption data. The study showed that the common practice of using linear regression needlessly limits the ability to model sorption data with good accuracy. Additional research resulted in a modified equation which addresses additional statistical limitations faced by researchers. For the soils tested in this study, the modified Langmuir equation provided better overall fits and generally more precise parameter estimates than the original Langmuir equation indicating that this approach is a more statistically valid method of obtaining sorption parameters with the Langmuir model. Because sorption parameters are often used to further our understanding of the underlying mechanisms controlling phosphorus sorption and for making management decisions regarding phosphorus application rates, it is imperative that these values are truly representative of the soil tested. This requires that the data be analyzed using proper statistical methods and that the data are fit with the correct sorption model. This research gives users improved regression methods for fitting models to sorption data. As part of this research several easy-to-use Microsoft Excel spreadsheets were developed capable of performing nonlinear regression and providing best-fit parameters and statistics of fit. The spreadsheets have been made available to the public via the World Wide Web and have been downloaded by over 150 researchers from more than 30 countries.

9. Broiler Litter Application Method and Runoff Timing Effects on Nutrient and E. Coli Losses from Tall Fescue Pasture. Runoff from agricultural land, particularly manure-applied lands is a major nonpoint source of nutrients, pathogenic microorganisms, and eroded sediment. ARS scientists from Bowling Green, KY, and Auburn, AL, used rainfall simulations to examine the effect of broiler litter application method and runoff timing on nutrient and E. Coli losses. Total phosphorus (TP), inorganic nitrogen, and E. Coli concentrations in runoff from traditional surface-broadcast litter application were all significantly greater than from subsurface litter banding. About 81% of the runoff TP was in the form of dissolved reactive phosphorus for both litter application methods. Increasing the time between litter application and the first runoff event helped decrease nutrient and E. Coli losses from surface-broadcast litter. It was documented that inorganic N and E. coli concentrations were significantly greater in runoff from plots receiving surface-broadcast applied poultry manure compared to plots where subsurface banding was used to apply the poultry manure. Subsurface litter banding into perennial grassland substantially reduce nutrient and pathogen losses in runoff compared to the traditional surface-broadcast practice.

10. Comparing Manure-derived Phosphorus Sorption with Inorganic Phosphorus Sorption. The application of phosphorus to agricultural fields can lead to pollution of nearby surface waters if the phosphorus is lost from the field and migrates to drinking water supplies. To mitigate this problem, the sorption behavior of phosphorus to soils is needed. In most phosphorus sorption studies phosphorus is added as an inorganic salt to a pre-defined background solution such as CaCl2 or KCl; however, in many regions the application of phosphorus to agricultural fields is in the form of animal manure. Scientists at Animal Waste Management Research Unit compared the sorption behavior of dissolved reactive phosphorus in KH2PO4 amended CaCl2 and KCl solutions with sorption behavior of dissolved reactive phosphorus in three different animal manure extracts. The results demonstrated that the sorption of phosphorus in animal manure solutions differs significantly from that of KH2PO4 amended CaCl2 and KCl solutions. This research shows that manure application rates based on sorption data collected from inorganic P salt experiments may be inaccurate. This is an important finding because the use of inorganic phosphorus salts for assessing sorption behavior of soils is widespread.

6.Technology Transfer

Number of Web Sites Managed1

Review Publications
Bolster, C.H., Sistani, K.R. 2009. Sorption of Phosphorus from Swine, Dairy, and Poultry Manures. Communications in Soil Science and Plant Analysis. 40:1106-1123

Warren, J.G., Phillips, S.B., Mullins, G.L. 2008. Impact of Soil Type and Application Rate on the Availability of P Applied as Alum-Treated Poultry Litter. Communications in Soil Science and Plant Analysis. Volume 39, Numbers 17-18, October 2008 , pp. 2515-2533(19)

Sistani, K.R., Torbert III, H.A., Way, T.R., Bolster, C.H., Pote, D.H., Warren, J.G. 2009. Broiler Litter Application Method and Runoff Timing Effects on Nutrient and Escherichia coli Losses from Tall Fescue Pasture. Journal of Environmental Quality. 38(3):1216-1223

Sistani, K.R., Mclaughlin, M.R., Brink, G.E. 2008. Soil nutrient evaluation from swine effluent application to five forage-system practices. Nutrient Cycling in Agroecosystems. 82:265-271.

Lovanh, N.C., Loughrin, J.H., Cook, K.L., Rothrock Jr, M.J., Sistani, K.R. 2009. The Effect of Stratification and Seasonal Variability on the Profile of an Anaerobic Swine Waste Treatment Lagoon. Bioresource Technology. 100:3706-3712

Loughrin, J.H., Lovanh, N.C., Arturo, Q., Rezaul, M. 2008. Sampling of Malodorous Compounds in Air Using Stir Bar Sorbtive Extraction. Transactions of the ASABE. Vol. 51(5): 1747-1752

Cook,K.L., Rothrock Jr,M.J., Warren,J.G., Sistani,K.R., Moore Jr,P.A. 2008. Effect of Alum Treatment on the Concentration of Total and Ureolytic Microorganisms in Poultry Litter. Journal of Environmental Quality. 37:2360-2367

Warren, J.G., Sistani, K.R., Way, T.R., Mays, D.A., Pote, D.H. 2008. A new method of poultry litter application to perennial pasture: subsurface banding. Soil Science Society of America Journal. 72: 1831-1837.

Tewolde, H., Armstrong, S., Way, T.R., Rowe, D., Sistani, K.R. 2009. Cotton response to poultry litter applied by subsurface banding relative to surface broadcasting. Soil Science Society of America Journal. 73:384-389.

Doerner, K., Mason, B., Kridelbaugh, D., Loughrin, J.H. 2009. Fe(III) stimulates 3-methylindole and 4-methylphenol production in swine lagoon enrichments and Clostridium scatologenes ATCC 25775. Letters in Applied Microbiology. 48 118-124

Mankolo, R.N., Sistani, K.R., Senwo, Z.N., Ranatunga, T.D. 2008. Seasonal changes in phosphorus and phosphatase compositions in soils enriched with poultry litter. Journal of Food Agriculture and Environment. Vol.6 (3&4):415-420 July-October 2008

Bolster, C.H. 2008. Revisiting a Statistical Shortcoming When Fitting the Langmuir Model to Sorption Data. Journal of Environmental Quality. Vol. 37 p. 1986-1992

Bolster, C.H., Haznedaroglu, B.Z., Walker, S.L. 2009. Diversity of Cell Properties and Transport Behavior Among 12 Enviromental Escherichia Coli Isolates. Journal of Environmental Quality. Vol. 38 p. 465-472

Bolster, C.H., Cook, K.L., Haznedaroglu, B.Z., Walker, S.L. 2009. The Transport of Mycobacterium avium subsp. paratuberculosis through Saturated Aquifer Materials. Letters in Applied Microbiology. vol. 48 p 307-312

Rothrock Jr, M.J., Cook, K.L., Warren, J.G., Sistani, K.R. 2008. The effect of alum addition on microbial communities in poultry litter. Poultry Science. 87(8) 1493-1503

Rothrock Jr, M.J., Cook, K.L., Bolster, C.H. 2009. Comparative quantification of Campylobacter jejuni from environmental samples using traditional and molecular biological techniques. Canadian Journal of Microbiology. 55: 633–641

Loughrin, J.H., Lovanh, N.C., Mahmood, R. 2008. Equilibrium Sampling Used to Monitor Malodors in a Swine Waste Lagoon. Journal of Environmental Quality. 37:1-6

Shahsavani, S., Ardalan, M., Sistani, K.R. 2006. Sulphate Adsorption in Soils of North and Northeast Iran.. Communications in Soil Science and Plant Analysis. 37:1587-1596.

Last Modified: 4/23/2014
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