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Influence of Swine Manure Application Method on Concentrations of Methanogens and Denitrifiers in Agricultural Soils
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Influence of Swine Manure Application Method on Concentrations of Methanogens and Denitrifiers in Agricultural Soils


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Soil microbial communities have been proposed as indicators of soil quality due to their importance as drivers of global biogeochemical cycles and their sensitivity to management and climatic conditions. Despite the importance of the soil microbiota to nutrient transformation and chemical cycling, physio-chemical properties rather than biological properties of soils are traditionally used as measures of environmental status. In general, much is unknown regarding the effect of management fluctuations on important functional groups in soils systems (i.e., methanogens, nitrifiers and denitrifiers). It is only recently that it has been possible, through application of sophisticated molecular microbiological methods, to sensitively and specifically target important microbial populations that contribute to nutrient cycling and plant health present at the field-scale and in differentially managed soil systems.

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  • We sampled 2 weeks after application – 4 days before the peak, so the microbial population should be responding to the application in distinguishable ways At the time of sampling Nitrifiers oxidize ammonium to nitrate under aerobic conditions Denitrifying bacteria reduce nitrate to n2o or n2 gas under anaerobic conditions N2O produced from NH2OH (hydroxylamine) or from nitrite reduction Ammonia and methan-oxidizing organisme produce n20 during the oxidation of hydroxylamine to nitrite AOB also reduce NO2- to N2O and N2 under anoxic conditions – through denitrification Nitrate reduction can be performed with three different purposes: the utilization of nitrate as a nitrogen source for growth (nitrate assimilation), the generation of metabolic energy by using nitrate as a terminal electron acceptor (nitrate respiration), and the dissipation of excess reducing power for redox balancing (nitrate dissimilation). IN absense of oxygen nitrifiers (facultative anaerobes) may switch to respiring nitrate
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    • 1. Influence of Swine Manure ApplicationInfluence of Swine Manure ApplicationMethod on Concentrations of MethanogensMethod on Concentrations of Methanogensand Denitrifiers in Agricultural Soilsand Denitrifiers in Agricultural SoilsKimberly Cook & Karamat SistaniKimberly Cook & Karamat SistaniBowling Green, KYBowling Green, KY
    • 2. Microbes & Biogeochemical CyclesMicrobes & Biogeochemical Cycles Indicators of soil qualityIndicators of soil quality Drivers of global biogeochemical cyclesDrivers of global biogeochemical cycles Sensitive to soil managementSensitive to soil management Sensitive to climate conditionsSensitive to climate conditions Physical vs BiologicalPhysical vs Biological Traditional measures focused on physio-chemicalTraditional measures focused on physio-chemical Molecular methods contribute to microbial analysesMolecular methods contribute to microbial analyses Better understand relationship between physical,Better understand relationship between physical,chemical and biological processes in agriculturalchemical and biological processes in agriculturalsystemssystems
    • 3. Microbes & GHGMicrobes & GHG Major Ag associated GHG: COMajor Ag associated GHG: CO22,,CHCH44 & N& N22OO Bacteria are largest producers of methane andBacteria are largest producers of methane andNN22OO Methane productionMethane production Complex anaerobic consortiaComplex anaerobic consortia Fermentative bacteria & methanogenic archaeaFermentative bacteria & methanogenic archaea Livestock and soilsLivestock and soils Anaerobic manure management systemsAnaerobic manure management systems NN22O from denitrification (anaerobic) andO from denitrification (anaerobic) andnitrification (anaerobic)nitrification (anaerobic)
    • 4. Cause & EffectCause & Effect How does management or environment affectHow does management or environment affectGHG production?GHG production? How does management or environment affectHow does management or environment affectbacteria that producebacteria that produce GHG?GHG?
    • 5. Swine Effluent Application StudySwine Effluent Application Study Evaluate the effect of pre-plant swine effluentEvaluate the effect of pre-plant swine effluentapplication method on GHG (COapplication method on GHG (CO22 ,CH,CH44 & N& N22O)O)emissions from soil in a no-till corn grainemissions from soil in a no-till corn grainproduction systemproduction system Sistani, K.R., Warren, J.G., Lovanh, N.C.,Sistani, K.R., Warren, J.G., Lovanh, N.C.,Higgins, S., Shearer, S. 2010. Green House GasHiggins, S., Shearer, S. 2010. Green House GasEmissions from Swine Effluent Applied to SoilEmissions from Swine Effluent Applied to Soilby Different Methods. Soil Sci. America J. 74(2):by Different Methods. Soil Sci. America J. 74(2):429-435.429-435.
    • 6. MethodsMethods Two growing seasons of no-till cornTwo growing seasons of no-till corn Gases measured using vented chambersGases measured using vented chambers Three effluent application methodsThree effluent application methods SurfaceSurface Direct InjectionDirect Injection Plus AerationPlus Aeration Microbiology – one sample point 2 weeks afterMicrobiology – one sample point 2 weeks afterapplication in 2008application in 2008
    • 7. InjectionInjectionSurface ApplicationSurface ApplicationAerationAerationMethods ofMethods ofSwine ManureSwine ManureApplicationApplication
    • 8.  COCO22 production similarproduction similarin all treatmentsin all treatments No influence of swineNo influence of swineeffluent applicationeffluent application
    • 9.  Total bacterial cell numbers similar in all treatmentsTotal bacterial cell numbers similar in all treatments Total fungal cell numbers similar in all treatmentsTotal fungal cell numbers similar in all treatments Averaged 3 orders of magnitude lower than total cellsAveraged 3 orders of magnitude lower than total cells
    • 10.  CHCH44 higher in injectionhigher in injectiontreatmenttreatment Emissions spiked for up toEmissions spiked for up to11 days after application11 days after application
    • 11.  Concentrations of methanogens (Concentrations of methanogens (mcrAmcrA) were up to) were up to7 orders of higher than background7 orders of higher than background Methane oxidizers (Methane oxidizers (pmoApmoA) were up to 6 orders of) were up to 6 orders ofmagnitude highermagnitude higher
    • 12.  NN22O flux highest in injectionO flux highest in injectiontreatmenttreatment Peak in flux at 6 dPeak in flux at 6 d However, aeration & injectionHowever, aeration & injectiontreatments continued totreatments continued toincrease & peaked after 18 dincrease & peaked after 18 d
    • 13. NitrifiersNitrifiers
    • 14. DenitrifiersDenitrifiersNitrate Reducers;Nitrate Reducers; narGnarGNitrite Reducers;Nitrite Reducers; nirKnirKNitrous Oxide Reducers;Nitrous Oxide Reducers; nosZnosZ
    • 15. Nitrate Reducers;Nitrate Reducers; narGnarG
    • 16. ConclusionsConclusions Two groups showed significant response toTwo groups showed significant response toeffluent applicationeffluent application Nitrifying Bacteria (AOB)Nitrifying Bacteria (AOB) Nitrate Reducers (Nitrate Reducers (narGnarG)) Methanogens and methane oxidizers alsoMethanogens and methane oxidizers alsoincreased orders of magnitude but were broughtincreased orders of magnitude but were broughtin with slurryin with slurry Sampling method, targeted genes and analysisSampling method, targeted genes and analysismethod are all significantmethod are all significant
    • 17. Integrating MicrobiologyIntegrating Microbiology Integrate into field analyses; incorporateIntegrate into field analyses; incorporatethinking into management practicesthinking into management practices Biological perspectiveBiological perspective Enzyme dependent pHEnzyme dependent pH Requirement for co-factorsRequirement for co-factors Mitigation of NMitigation of N22OO in atmosphere exclusivelycarried out by nitrous oxide reducers (nosZ)nitrous oxide reducers (nosZ) Requires 12 Copper ionsRequires 12 Copper ions Optimum pH over 7Optimum pH over 7
    • 18. CollaboratorsCollaborators Special thanks to Dr. Jason Warren (OK StateSpecial thanks to Dr. Jason Warren (OK StateUniversity) for collaborative efforts on thisUniversity) for collaborative efforts on thisprojectproject Thanks also to Rohan Parekh and JasonThanks also to Rohan Parekh and JasonSimmons for technical assistanceSimmons for technical assistanceThis research was conducted as part of USDA-This research was conducted as part of USDA-ARS National Program 214: Agricultural andARS National Program 214: Agricultural andIndustrial By-ProductsIndustrial By-Products