Assessing the Impact of Non-Urea Ruminant Urine Nitrogen Compounds on Urine Patch Nitrous Oxide Emissions.

Urea, the dominant form of N in ruminant urine, degrades in soil to produce NO emissions. However, the fate of non-urea urine N compounds (NUNCs) in soil and their contribution to urine patch NO emissions remain unclear. This study evaluated five NUNCs: allantoin (10%), creatinine (3%), creatine (3%), uric acid (1%), and (hypo)xanthine (0.6%), where numbers in parentheses represent the average percentage of total urine N. The fates of NUNCs in a pasture soil were determined using N-labeled NUNCs in a laboratory trial. Two NUNCs, hypoxanthine and creatine, were added to the soil with perennial ryegrass ( L.) present and sampled over time for soil inorganic N, NO emissions, and plant N dynamics. The N enrichments of soil inorganic N and plant N were significantly increased within 24 h of NUNC application, indicating rapid microbial degradation and plant uptake of NUNCs in pasture soil. An autumn field trial was also conducted to evaluate the in situ impact of varying concentrations of NUNCs on urine patch NO emissions. Increasing the proportion of urine N excreted as NUNCs did not alter the urine patch NO emission factor, soil inorganic N concentrations, or plant N uptake. It is concluded that NUNCs rapidly degrade in pasture soil and that an increased ruminant excretion of urine N as NUNCs does not significantly alter the urine patch NO emission factor.

Transformation of Organic Matter and the Emissions of Methane and Ammonia during Storage of Liquid Manure as Affected by Acidification.

Acidification of livestock manure can reduce emission of the greenhouse gases methane (CH) and nitrous oxide (NO), as well as ammonia (NH). We examined the relation between emission of these gases and transformation of organic matter as affected by acidification. Liquid cattle manure was acidified with sulfuric acid to pH 5.5 at a pilot scale (100 L), and we measured effects on CH, NO, CO and NH emissions and on transformation of pH buffer components and organic matter. Acidification reduced NH emissions by 62% (47 d) and emission of CH by 68% (57 d). Emissions of NO were negligible, probably due to the absence of a surface crust. Reductions in NH and CH emission were highest at the start but declined over time concomitantly with a gradual increase in the stored liquid manure pH. Acidification did not significantly affect CO emissions. Emission of CO was high, five- to ten-fold of CH emissions, until Day 16 of storage, after which the CO emission rate declined to around twice the CH emission rate; consequently, the majority of C loss during the early stages of storage was CO. Cumulative emission of C in CO and CH closely matched depletion of dissolved organic carbon (DOC), suggesting that DOC may be a predictor for CH emission from dilute slurries. volatile fatty acid and total ammoniacal nitrogen concentrations in surface layers were substantially higher than at the center of stored liquid manure, perhaps resulting from microbial activity at the surface. This pattern deserves attention when predicting NH emission from stored slurry.

The response of ammonia-oxidizing microorganisms to trace metals and urine in two grassland soils in New Zealand.

An incubation experiment was conducted to investigate the response of ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA), and the nitrification rate to the contamination of Cu, Zn, and Cd in two New Zealand grassland soils. The soils spiked with different concentrations of Cu (20 and 50 mg kg-1), Zn (20 and 50 mg kg-1), and Cd (2 and 10 mg kg-1) were incubated for 14 days and then treated with 500 mg kg-1 urine-N before continuing incubation for a total of 115 days. Soils were sampled at intervals throughout the incubation. The nitrification rate in soils at each sampling period was determined, and the abundance of AOB and AOA was measured by real-time quantification polymerase chain reaction (qPCR) assay of the amoA gene copy numbers. The results revealed that moderate trace metal stress did not significantly affect the abundance of AOB and AOA in the two soils, probably due to the high organic matter content of the soils which would have reduced the toxic effect of the metals. Nitrification rates were much greater and the observable nitrification period was much shorter in the dairy farm (DF) soil, in which the AOB and AOA abundances were greater than those of the mixed cropping farm (MF) soil. AOB were shown to grow under high nitrogen conditions, whereas AOA were shown to grow under low N environments, with different metal concentrations. Therefore, nitrogen status rather than metal applications was the main determining factor for AOB and AOA growth in the two soils studied.

Confirmation of co-denitrification in grazed grassland.

Pasture-based livestock systems are often associated with losses of reactive forms of nitrogen (N) to the environment. Research has focused on losses to air and water due to the health, economic and environmental impacts of reactive N. Di-nitrogen (N2) emissions are still poorly characterized, both in terms of the processes involved and their magnitude, due to financial and methodological constraints. Relatively few studies have focused on quantifying N2 losses in vivo and fewer still have examined the relative contribution of the different N2 emission processes, particularly in grazed pastures. We used a combination of a high (15)N isotopic enrichment of applied N with a high precision of determination of (15)N isotopic enrichment by isotope-ratio mass spectrometry to measure N2 emissions in the field. We report that 55.8 g N m(-2) (95%, CI 38 to 77 g m(-2)) was emitted as N2 by the process of co-denitrification in pastoral soils over 123 days following urine deposition (100 g N m(-2)), compared to only 1.1 g N m(-2) (0.4 to 2.8 g m(-2)) from denitrification. This study provides strong evidence for co-denitrification as a major N2 production pathway, which has significant implications for understanding the N budgets of pastoral ecosystems.

Effects of nitrogen application rate and a nitrification inhibitor dicyandiamide on methanotroph abundance and methane uptake in a grazed pasture soil.

Methane-oxidizing bacteria (methanotrophs) in the soil are a unique group of methylotrophic bacteria that utilize methane (CH4) as their sole source of carbon and energy which limit the flux of methane to the atmosphere from soils and consume atmospheric methane. A field experiment was conducted to determine the effect of nitrogen application rates and the nitrification inhibitor dicyandiamide (DCD) on the abundance of methanotrophs and on methane flux in a grazed pasture soil. Nitrogen (N) was applied at four different rates, with urea applied at 50 and 100 kg N ha(-1) and animal urine at 300 and 600 kg N ha(-1). DCD was applied at 10 kg ha(-1). The results showed that both the DNA and selected mRNA copy numbers of the methanotroph pmoA gene were not affected by the application of urea, urine or DCD. The methanotroph DNA and mRNA pmoA gene copy numbers were low in this soil, below 7.13 × 10(3) g(-1) soil and 3.75 × 10(3) µg(-1) RNA, respectively. Daily CH4 flux varied slightly among different treatments during the experimental period, ranging from -12.89 g CH4 ha(-1) day(-1) to -0.83 g CH4 ha(-1) day(-1), but no significant treatment effect was found. This study suggests that the application of urea fertilizer, animal urine returns and the use of the nitrification inhibitor DCD do not significantly affect soil methanotroph abundance or daily CH4 fluxes in grazed grassland soils.

Effects of nitrogen application rate and a nitrification inhibitor dicyandiamide on ammonia oxidizers and N2O emissions in a grazed pasture soil.

Ammonia oxidizers, including ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA) are important drivers of a key step of the nitrogen cycle - nitrification, which affects the production of the potent greenhouse gas, nitrous oxide (N2O). A field experiment was conducted to determine the effect of nitrogen application rates and the nitrification inhibitor dicyandiamide (DCD) on the abundance of AOB and AOA and on N2O emissions in a grazed pasture soil. Nitrogen (N) was applied at four different rates, with urea applied at 50 and 100 kg N ha(-1) and animal urine at 300 and 600 kg N ha(-1). DCD was applied to some of the N treatments at 10 kg ha(-1). The results showed that the AOB amoA gene copy numbers were greater than those of AOA. The highest ratio of the AOB to AOA amoA gene copy numbers was 106.6 which occurred in the urine-N 600 treatment. The AOB amoA gene copy numbers increased with increasing nitrogen application rates. DCD had a significant impact in reducing the AOB amoA gene copy numbers especially in the high nitrogen application rates. N2O emissions increased with the N application rates. DCD had the most significant effect in reducing the daily and total N2O emissions in the highest nitrogen application rate. The greatest reduction of total N2O emissions by DCD was 69% in the urine-N 600 treatment. The reduction in the N2O emission factor by DCD ranged from 58% to 83%. The N2O flux and NO3(-)-N concentrations were significantly correlated to the growth of AOB, rather than AOA. This study confirms the importance of AOB in nitrification and the effect of DCD in inhibiting AOB growth and in decreasing N2O emissions in grazed pasture soils under field conditions.

Ammonia-oxidizing bacteria and archaea grow under contrasting soil nitrogen conditions.

Nitrification is a key process of the nitrogen (N) cycle in soil with major environmental implications. The recent discovery of ammonia-oxidizing archaea (AOA) questions the traditional assumption of the dominant role of ammonia-oxidizing bacteria (AOB) in nitrification. We investigated AOB and AOA growth and nitrification rate in two different layers of three grassland soils treated with animal urine substrate and a nitrification inhibitor [dicyandiamide (DCD)]. We show that AOB were more abundant in the topsoils than in the subsoils, whereas AOA were more abundant in one of the subsoils. AOB grew substantially when supplied with a high dose of urine substrate, whereas AOA only grew in the Controls without the urine-N substrate. AOB growth and the amoA gene transcription activity were significantly inhibited by DCD. Nitrification rates were much higher in the topsoils than in the subsoils and were significantly related to AOB abundance, but not to AOA abundance. These results suggest that AOB and AOA prefer different soil N conditions to grow: AOB under high ammonia (NH(3)) substrate and AOA under low NH(3) substrate conditions.

Transport and modeling of estrogenic hormones in a dairy farm effluent through undisturbed soil lysimeters.

The presence of endocrine-disrupting chemicals, including estrone (E1) and 17beta-estradiol (E2), in surface waters has been associated with physiological dysfunction in a number of aquatic organisms. One source of surface and groundwater contamination with E1 and E2 is the land application of animal wastes. The processes involved in the transport of these hormones in the soil, when applied with animal wastes, are still unclear. Therefore, a field-transport experiment was carried out, where a dairy farm effluent spiked with E1 and E2 was applied on large (50 cm diameter and 70 cm depth) undisturbed soil lysimeters. The concentrations of E1 and E2 in the leachate were monitored over a 3-month period, during which irrigation was applied. The experimental data suggest that E1 and E2 were transported through preferential/macropore flow pathways. The data from the experiment also show that E1 and E2 are leached earlier than the inert tracer (bromide). This observation can be explained either by the presence of antecedent concentrations in the soil or by an enhanced transport of E1 and E2 through the soil. A state-space mixing-cell model was further developed in order to describe the transport of E1 and E2 by three transport processes in parallel. The inverse modeling of the leaching data did not support the hypothesis that antecedent concentrations of estrogens could be responsible for the observed breakthrough curves but confirmed that estrogens were transported mainly via preferential/macropore flow and also via an enhanced transport. The parameter values that characterized this enhanced transport strongly suggest that this enhanced transport is mediated by colloids. For the first time, the simultaneous transport of E1 and E2 was modeled under transient conditions, taking into account the advection-dispersion, preferential/macropore flow, and colloidal-enhanced transport processes as well as E1 and E2 dissipation in the soil. These findings have major implications in terms of management practices to decrease E1 and E2 transport and water contamination.

Sorption of estrone and estrone-3-sulfate from CaCl2 solution and artificial urine in pastoral soils of New Zealand.

Estrone (E1) and its sulfate conjugate estrone-3-sulfate (E1-3S) are released to the environment in animal wastes in significant amounts, and direct exposure occurs in grazed pasture systems. Both compounds have been shown to potentially contribute to endocrine disruption in wildlife, and knowledge about the sorption behavior of these compounds is necessary for a sound risk assessment. For labile compounds such as E1 and E1-3S, however, the standard protocols might overestimate sorption by not considering metabolite formation or allowing for equilibration that exceeds the commonly reported half-lives of these compounds. We therefore conducted modified batch sorption experiments with 0.005 M calcium chloride (CaCl2) and artificial urine solution to determine the influence of the mediator solution on the sorption of E1 and E1-3S in three pasture soils from New Zealand. Sorption isotherms of both compounds were nonlinear, and the Freundlich equation was found adequate to describe the isotherms. The sorption potential of E1-3S was about one order of magnitude lower than for the free counterpart, and the Kf values significantly changed between the two mediator solutions. The calculation of concentration-dependent effective distribution coefficients (Kdeff) revealed that for a range of realistic exposure concentrations in a grazed dairy system, the common approach of using CaCl2 would deliver incorrect inferences for a sound risk assessment.

Degradation and metabolite formation of 17beta-estradiol-3-sulphate in New Zealand pasture soils.

Estrogens-sulphates such as 17beta-estradiol-3-sulphate and estrone-3-sulphate are excreted by livestock in the urine. These conjugates are precursors to the free counterparts 17beta-estradiol and estrone, which are endocrine disrupting chemicals. In this study microcosm laboratory experiments were conducted in three pasture soils from New Zealand to study the aerobic degradation and metabolite formation kinetics of 17beta-estradiol-3-sulphate at three different incubation temperatures. The degradation of 17beta-estradiol-3-sulphate followed a first-order kinetic and the temperature dependence of the rate constants was sufficiently described by the Arrhenius equation. Degradation was different between the three investigated soils and the rate constants across the soils were significantly correlated to the arylsulphatase activity at 7.5 and 15 degrees C. Estrone-3-sulphate and 17beta-estradiol were identified as primary metabolites and estrone as a secondary metabolite. Results suggest arylsulphatase activity originating from soil microbial biomass is the main driver for the degradation of 17beta-estradiol-3-sulphate.

Modeling degradation and metabolite formation kinetics of estrone-3-sulfate in agricultural soils.

Estrone-3-sulfate (E1-3S), formed in the kidneys of pregnant cattle, can act as a precursor to the free hormone estrone (E1) known for its endocrine disrupting potential in wildlife. Laboratory microcosm studies were conducted to investigate the aerobic degradation of E1-3S in three contrasting pasture soils at 7.5, 15, and 25 degrees C. Deconjugation of E1-3S resulted in the formation of the metabolite E1. Two kinetic models-a single first-order and a biexponential kinetic model-were applied to fit the observed degradation dynamics and to derive degradation end-points (DT50 and DT90) for the parent compound and the metabolite for each condition. Model selection and evaluation of their performance were based on a suit of statistical measures (one-way ANOVA, AIC(c), R2(adj), chi2 error-%, and SRMSE). The results showed rapid initial degradation of E1-3S, followed by a much slower decline with time, and rate of degradation was temperature dependent. The DT50 and DT90 values of E1-3S ranged from a few hours to several days, while the formation of the major metabolite (E1) was concomitant with E1-3S degradation in all nonsterile soils. The parent compound degradation and formation and subsequent dissipation of metabolite were successfully predicted by both models, however, the nonlinear biexponential model improved the goodness-of-fit parameters in most cases.

Fungal inoculum properties: extracellular enzyme expression and pentachlorophenol removal in highly contaminated field soils.

This study was conducted to improve the pentachlorophenol (PCP) bioremediation ability of white-rot fungi in highly contaminated field soils by manipulating bioaugmentation variables. These were the dry weight percentage of fungal inoculum addition (31-175 g kg(-1)), PCP concentration (100-2137 mg kg(-1) PCP), fungal inoculum formulation, and time (1-7 wk). Five fungal isolates were used: the New Zealand isolates Trametes versicolor (L.: Fr.) HR131 and Trametes sp. HR577; the North American isolates Phanerochaete chrysosporium Burds. (two isolates) and Phanerochaete sordida (Karst.) Erikss. & Ryv. Pentachlorophenol removal, manganese peroxidase, and laccase activity, and the formation of chloroanisoles in the contaminated field soils were measured. The majority of PCP removed by the Trametes isolates was in the first week after bioaugmentation. The maximum PCP removal by the fungi varied from 50 to 65% from a 1065 mg kg(-1) PCP contaminated field soil. Pentachlorophenol was preferentially converted to pentachloroanisole (PCA) by the Phanerochaete isolates (>60%), while 2 to 9% of the PCP removed by two Trametes isolates was converted to PCA. A pH increase was measured following bioaugmentation that was dependent on PCP concentration, fungal inoculum addition, and formulation. This, together with rapid initial PCP removal, possibly changed the bioavailability of the remaining PCP to the fungi and significantly decreased the sequestering of PCP in the contaminated field soils. The research supports the conclusion that New Zealand Trametes spp. can rapidly remove PCP in contaminated field soils. Bioavailability and extractability of PCP in the contaminated field soil may significantly increase after bioaugmentation.

Fungal inoculum properties: extracellular enzyme expression and pentachlorophenol removal by New Zealand trametes species in contaminated field soils.

This study was conducted to improve the ability of indigenous New Zealand white-rot fungi to remove pentachlorophenol (PCP) from contaminated field soil. The effects of different bioaugmentation conditions on PCP removal and extracellular enzyme expression were measured in the laboratory. The conditions were fungal growth substrate and co-substrate composition, culture age, and Tween 80 addition to the contaminated soil. The fungi used were Trametes versicolor isolate HR131 and Trametes sp. isolate HR577. Maximum PCP removal was 70% after 7 wk from a 1043 mg kg(-1) PCP-contaminated soil inoculated with an 11-d-old fungal culture of T. versicolor isolate HR131. There was minimal production of undesirable pentachloroanisole by the fungi. Tween 80 addition had no affect on PCP removal. Poplar sawdust was more suitable as a fungal growth substrate and a co-substrate amendment for PCP removal and extracellular enzyme expression than the locally available pine and fir sawdust. Pentachlorophenol removal was not necessarily correlated with extracellular enzyme expression. The research results demonstrate that PCP biodegradation was affected by inoculum culture age, by the presence of a co-substrate amendment, and by growth substrate composition after white-rot fungal bioaugmentation into PCP-contaminated field soils.

Ammonia, methane, and nitrous oxide emission from pig slurry applied to a pasture in New Zealand.

Much animal manure is being applied to small land areas close to animal confinements, resulting in environmental degradation. This paper reports a study on the emissions of ammonia (NH3), methane (CH4), and nitrous oxide (N2O) from a pasture during a 90-d period after pig slurry application (60 m3 ha-1) to the soil surface. The pig slurry contained 6.1 kg total N m-3, 4.2 kg of total ammoniacal nitrogen (TAN = NH3 + NH4) m-3, and 22.1 kg C m-3, and had a pH of 8.14. Ammonia was lost at a fast rate immediately after slurry application (4.7 kg N ha-1 h-1), when the pH and TAN concentration of the surface soil were high, but the loss rate declined quickly thereafter. Total NH3 losses from the treated pasture were 57 kg N ha-1 (22.5% of the TAN applied). Methane emission was highest (39.6 g C ha-1 h-1) immediately after application, as dissolved CH4 was released from the slurry. Emissions then continued at a low rate for approximately 7 d, presumably due to metabolism of volatile fatty acids in the anaerobic slurry-treated soil. The net CH4 emission was 1052 g C ha-1 (0.08% of the carbon applied). Nitrous oxide emission was low for the first 14 d after slurry application, then showed emission peaks of 7.5 g N ha-1 h-1 on Day 25 and 15.8 g N ha-1 h-1 on Day 67, and decline depending on rainfall and nitrate (NO3) concentrations. Emission finally reached background levels after approximately 90 d. Nitrous oxide emission was 7.6 kg N ha-1 (2.1% of the N applied). It is apparent that of the two major greenhouse gases measured in this study, N2O is by far the more important tropospheric pollutant.