An analysis of the spatio-temporal occurrence of anthelmintic veterinary drug residues in groundwater.

Anthelmintics are antiparasitic drugs used to control helminthic parasites such as nematodes and trematodes in animals, particularly those exposed through pasture-based production systems. Even though anthelmintics have been shown to be excreted into the environment in relatively high amounts as unmetabolized drug or transformation products (TPs), there is still only limited information available on their environmental occurrence, particularly in groundwater, which has resulted in them being considered as potential emerging contaminants of concern. A comprehensive study was carried out to investigate the occurrence of 40 anthelmintic residues (including 13 TPs) in groundwaters (and associated surface waters) throughout the Republic of Ireland. The study focused on investigating the occurrence of these contaminants in karst and fractured bedrock aquifers, with a total of 106 sites (88 groundwaters and 18 surface waters) samples during spring 2017. Seventeen anthelmintic compounds consisting of eight parent drugs and nine TPs were detected at 22% of sites at concentrations up to 41 ng L-1. Albendazole and its TPs were most frequently detected residues, found at 8% of groundwater sites and 28% of surface water sites. Multivariate statistical analysis identified several source and pathway factors as being significantly related to the occurrence of anthelmintics in groundwater, however there was an evident localised effect which requires further investigation. An investigation of the temporal variations in occurrence over a 13 month period indicated a higher frequency and concentration of anthelmintics during February/March and again later during August/September 2018, which coincided with periods of increased usage and intensive meteorological events. This work presents the first detections of these contaminants in Irish groundwater and it contributes to broadening our understanding of anthelmintics in the environment. It also provides insight to seasonal trends in occurrence, which is critical for assessing potential future effects and implications of climate change.

An investigation of anticoccidial veterinary drugs as emerging organic contaminants in groundwater.

Intensification of the food production system to meet increased global demand for food has led to veterinary pharmaceuticals becoming a critical component in animal husbandry. Anticoccidials are a group of veterinary products used to control coccidiosis in food-producing animals, with primary prophylactic use in poultry production. Excretion in manure and subsequent land-spreading provides a potential pathway to groundwater. Information on the fate and occurrence of these compounds in groundwater is scant, therefore these substances are potential emerging organic contaminants of concern. A study was carried out to investigate the occurrence of anticoccidial compounds in groundwater throughout the Republic of Ireland. Twenty-six anticoccidials (6 ionophores and 20 synthetic anticoccidials) were analysed at 109 sites (63 boreholes and 46 springs) during November and December 2018. Sites were categorised and selected based on the following source and pathway factors: (a) the presence/absence of poultry activity (b) predominant aquifer category and (c) predominant groundwater vulnerability, within the zone of contribution (ZOC) for each site. Seven anticoccidials, including four ionophores (lasalocid, monensin, narasin and salinomycin) and three synthetic anticoccidials (amprolium, diclazuril and nicarbazin), were detected at 24% of sites at concentrations ranging from 1 to 386 ng L-1. Monensin and amprolium were the two most frequently detected compounds, detected at 15% and 7% of sites, respectively. Multivariate statistical analysis has shown that source factors are the most significant drivers of the occurrence of anticoccidials, with no definitive relationships between occurrence and pathway factors. The study found that the detection of anticoccidial compounds is 6.5 times more likely when poultry activity is present within the ZOC of a sampling point, compared to the absence of poultry activity. This work presents the first detections of these contaminants in Irish groundwater and it contributes to broadening our understanding of the environmental occurrence and fate of anticoccidial veterinary products.

Application of 15N tracing for estimating nitrogen cycle processes in soils of a constructed wetland.

Integrated Constructed Wetlands (ICW) area technology for the attenuation of contaminants such as organic carbon (C), nitrogen (N), phosphorous (P) and sulphur (S) in water coming from point or diffuse sources. Currently there is a lack of knowledge on the rates of gross N transformations in soils of the ICW bed leading to losses of reactive N to the environment. In addition, the kinetics of these processes need to be studied thoroughly for the sustainable use of ICW for removal of excessive N in the treatment of waste waters. Gross N transformation processes were quantified at two soil depths (0-15 and 30-45 cm) in the bed of a surface flow ICW using a 15N tracing approach. The ICW, located in Dunhill village at Waterford in Southeastern Ireland, receives 500 person equivalent waste waters containing large quantities of organic pollutants (ca. mean annual C, N, P and S contents of 240, 60, 5 and 73 mg L-1). Soil was removed from these depths in December 2014 and incubated anaerobically in the laboratory, with either 15N labeled ammonium (NH4+) or nitrate (NO3-), differentially labeled with 14NH415NO3 and 15NH414NO3 in parallel setups, enriched to 50 atm% 15N. Results showed that at both soil depths, NO3- production rates were small, which may have resulted in lower NO3- reduction by either denitrification or dissimilatory NO3- reduction to ammonium (DNRA). However, despite being low, the DNRA rates were greater than denitrification rates. Direct transformation of organic N to NO3-, without mineralization to NH4+, was a prevalent pathway of NO3- production accounting for 28-33% of the total NO3- production. Relative contribution of this process to the total N mineralization was negligible at depth 1 (0.01%) but dominant at depth 2 (99.7%). Total NO3-production to total immobilization of NH4+ and NO3- was very small (<0.50%) suggesting that ICW soils are not a source of NO3-. Despite a large potential of N immobilization existed at both the layers, relative N immobilization to the total N conversion was higher at depth 2 (ca. 2.2) than at depth 1 (ca. 1.5). The NH4+ desorption rate at 30-45 cm was high. However, immobilization in the recalcitrant and labile organic N pools was higher. Mineralization and immobilization of NH4+ processes showed that recalcitrant organic N was the predominant source in ICW soils whereas the labile organic N was comparatively small. Source apportionment of N2O production showed that the majority of the N2O produced through denitrification (ca. 92.5%) followed by heterotrophic nitrification (ca. 5.5%), co-denitrification (ca. 1.90%) and nitrification (0.20%). These results revealed that application of a detailed 15N tracing method can provide insights on the underlying processes of ecosystem based abundances of reactive N. A key finding of this study was that both investigated ICW layers were characterised by large N immobilization which restricts production of NO3- and further gaseous N losses.

Nitrogen fertilisers with urease inhibitors reduce nitrous oxide and ammonia losses, while retaining yield in temperate grassland.

Nitrogen fertilisation, although a cornerstone of modern agricultural production, negatively impacts the environment through gaseous losses of nitrous oxide (N2O), a potent greenhouse gas (GHG), and ammonia (NH3), a known air pollutant. The aim of this work was to assess the feasibility of urea treated with urease inhibitors to reduce gaseous N losses in temperate grassland, while maintaining or improving productivity compared to conventional fertiliser formulations. Urease inhibitors were N-(n-butyl)-thiophosphoric triamide (NBPT) (urea + NBPT) and N-(n-propyl)-thiophosphoric triamide (NPPT) (urea+ NBPT + NPPT), while conventional fertilisers were urea and calcium ammonium nitrate (CAN). N2O emission factors were 0.06%, 0.07%, 0.09% and 0.58% from urea + NBPT, urea, urea + NBPT + NPPT and CAN, respectively, with CAN significantly higher than all the urea formulations, which were not significantly different from each other. Ammonia loss measured over one fertiliser application was significantly larger from urea, at 43%, compared with other formulations at 13.9%, 13.8% and 5.2% from urea + NBPT, urea + NBPT + NPPT and CAN, respectively. Changing fertiliser formulation had no significant impact on grass yield or N uptake in four out of five harvests. In the last harvest urea + NBPT significantly out-yielded urea, but not CAN or urea + NBPT + NPPT. Overall, urea treated with either one or both urease inhibitors significantly reduced emissions of N2O and NH3, while preserving yield quantity and quality. Therefore, changing fertiliser formulation to these products should be encouraged as a strategy to reduce GHG and air pollution from agricultural practices in temperate climate.

A new sensitive method for the simultaneous chromatographic separation and tandem mass spectrometry detection of anticoccidials, including highly polar compounds, in environmental waters.

A sensitive and selective method was developed and validated for the determination of 26 anticoccidial compounds (six ionophores and twenty chemical coccidiostats) in surface and groundwater samples at parts-per-quadrillion (pg L-1) to parts-per-trillion (ng L-1) levels by ultra-high performance liquid chromatography with tandem mass spectrometry detection (UHPLC-MS/MS). A range of different analytical columns and mobile phase compositions were evaluated to enhance selectivity and retention of a number of highly polar and basic anticoccidials along with other non-polar coccidiostats. A combined separation, including these problematic polar compounds, was achieved on a phenyl-hexyl column, by binary gradient elution with water/acetonitrile using ammonium formate and formic acid as additives. The anticoccidial residues were extracted from raw, unfiltered, water samples (250 mL) using polymeric divinylbenzene solid phase extraction (SPE) cartridges, with subsequent elution (methanol:acetonitrile:ethyl acetate, 40:40:20, v/v) and concentration prior to determination. The method recovery (at a concentration representative of realistic expected environmental water concentrations based on literature review) ranged from 81% to 105%. The method was successfully validated for 26 anticoccidials, at four concentration levels, in accordance to Commission Decision 2002/657/EC and SANTE/11813/2017 guidelines. Trueness and precision, under within-laboratory reproducibility conditions, ranged from 88% to 111% and 0.9% to 10.3% respectively.

Scenarios to limit environmental nitrogen losses from dairy expansion.

Increased global demand for dairy produce and the abolition of EU milk quotas have resulted in expansion in dairy production across Europe and particularly in Ireland. Simultaneously, there is increasing pressure to reduce the impact of nitrogen (N) losses to air and groundwater on the environment. In order to develop grassland management strategies for grazing systems that meet environmental targets and are economically sustainable, it is imperative that individual mitigation measures for N efficiency are assessed at farm system level. To this end, we developed an excel-based N flow model simulating an Irish grass-based dairy farm, to evaluate the effect of farm management on N efficiency, N losses, production and economic performance. The model was applied to assess the effect of different strategies to achieve the increased production goals on N utilization, N loss pathways and economic performance at farm level. The three strategies investigated included increased milk production through increased grass production, through increased concentrate feeding and by applying a high profit grass-based system. Additionally, three mitigation measures; low ammonia emission slurry application, the use of urease and nitrification inhibitors and the combination of both were applied to the three strategies. Absolute N emissions were higher for all intensification scenarios (up to 124 kg N ha-1) compared to the baseline (80 kg N ha-1) due to increased animal numbers and higher feed and/or fertiliser inputs. However, some intensification strategies showed the potential to reduce the emissions per ton milk produced for some of the N-loss pathways. The model showed that the assessed mitigation measures can play an important role in ameliorating the increased emissions associated with intensification, but may not be adequate to entirely offset absolute increases. Further improvements in farm N use efficiency and alternatives to mineral fertilisers will be required to decouple production from reactive N emissions.

Feeding dicyandiamide (DCD) to cattle: An effective method to reduce N2O emissions from urine patches in a heavy-textured soil under temperate climatic conditions.

Nitrate (NO3-) leaching and nitrous oxide (N2O) emission from urine patches in grazed pastures are key sources of water and air pollution, respectively. Broadcast spraying of the nitrification inhibitor dicyandiamide (DCD) has been shown to reduce these losses, but it is expensive. As an alternative, it had been demonstrated that feeding DCD to cattle (after manual mixing with supplementary feeds) was a practical, effective and cheaper method to deliver high DCD rates within urine patches. This two-year study carried out on simulated urine patches in three application seasons (spring, summer, autumn) explored the efficacy of DCD feeding to cattle to reduce N losses from grazed pasture soil in a heavy-textured soil under temperate climatic conditions. In each application season, DCD fed to cows, then excreted with urine and applied at a rate of 30kgDCDha-1 (treatment U+DCD30-f) was as effective as powdered DCD mixed with normal urine and applied at the same rate (treatment U+DCD30) at reducing cumulative N2O-N emissions and the N2O-N emission factor (EF3, expressed as % of N applied). Increasing DCD loading within urine patches from 10 to 30kgDCDha-1 improved efficacy by significantly reducing the EF3 from 34% to 64%, which highlights that under local conditions, 10kgDCDha-1 (the recommended rate for commercial use in New Zealand) was not the optimum DCD rate to curb N2O emissions. The modelling of EF3 in this study also suggests that N mitigation should be given more attention when soil moisture is going to be high, which can be predicted with short-term weather forecasting. DCD feeding, for instance in autumn when cows are not lactating and the risk of N losses is high, could then be reduced by focusing mainly on those forecasted wet periods.

Combining stable isotopes with contamination indicators: A method for improved investigation of nitrate sources and dynamics in aquifers with mixed nitrogen inputs.

Excessive nitrate (NO3-) concentration in groundwater raises health and environmental issues that must be addressed by all European Union (EU) member states under the Nitrates Directive and the Water Framework Directive. The identification of NO3- sources is critical to efficiently control or reverse NO3- contamination that affects many aquifers. In that respect, the use of stable isotope ratios 15N/14N and 18O/16O in NO3- (expressed as δ15N-NO3- and δ18O-NO3-, respectively) has long shown its value. However, limitations exist in complex environments where multiple nitrogen (N) sources coexist. This two-year study explores a method for improved NO3- source investigation in a shallow unconfined aquifer with mixed N inputs and a long established NO3- problem. In this tillage-dominated area of free-draining soil and subsoil, suspected NO3- sources were diffuse applications of artificial fertiliser and organic point sources (septic tanks and farmyards). Bearing in mind that artificial diffuse sources were ubiquitous, groundwater samples were first classified according to a combination of two indicators relevant of point source contamination: presence/absence of organic point sources (i.e. septic tank and/or farmyard) near sampling wells and exceedance/non-exceedance of a contamination threshold value for sodium (Na+) in groundwater. This classification identified three contamination groups: agricultural diffuse source but no point source (D+P-), agricultural diffuse and point source (D+P+) and agricultural diffuse but point source occurrence ambiguous (D+P±). Thereafter δ15N-NO3- and δ18O-NO3- data were superimposed on the classification. As δ15N-NO3- was plotted against δ18O-NO3-, comparisons were made between the different contamination groups. Overall, both δ variables were significantly and positively correlated (p < 0.0001, rs = 0.599, slope of 0.5), which was indicative of denitrification. An inspection of the contamination groups revealed that denitrification did not occur in the absence of point source contamination (group D+P-). In fact, strong significant denitrification lines occurred only in the D+P+ and D+P± groups (p < 0.0001, rs > 0.6, 0.53 ≤ slope ≤ 0.76), i.e. where point source contamination was characterised or suspected. These lines originated from the 2-6‰ range for δ15N-NO3-, which suggests that i) NO3- contamination was dominated by an agricultural diffuse N source (most likely the large organic matter pool that has incorporated 15N-depleted nitrogen from artificial fertiliser in agricultural soils and whose nitrification is stimulated by ploughing and fertilisation) rather than point sources and ii) denitrification was possibly favoured by high dissolved organic content (DOC) from point sources. Combining contamination indicators and a large stable isotope dataset collected over a large study area could therefore improve our understanding of the NO3- contamination processes in groundwater for better land use management. We hypothesise that in future research, additional contamination indicators (e.g. pharmaceutical molecules) could also be combined to disentangle NO3- contamination from animal and human wastes.

Groundwater nitrate reduction versus dissolved gas production: A tale of two catchments.

At the catchment scale, a complex mosaic of environmental, hydrogeological and physicochemical characteristics combine to regulate the distribution of groundwater and stream nitrate (NO3-). The efficiency of NO3- removal (via denitrification) versus the ratio of accumulated reaction products, dinitrogen (excess N2) & nitrous oxide (N2O), remains poorly understood. Groundwater was investigated in two well drained agricultural catchments (10km2) in Ireland with contrasting subsurface lithologies (sandstone vs. slate) and landuse. Denitrification capacity was assessed by measuring concentration and distribution patterns of nitrogen (N) species, aquifer hydrogeochemistry, stable isotope signatures and aquifer hydraulic properties. A hierarchy of scale whereby physical factors including agronomy, water table elevation and permeability determined the hydrogeochemical signature of the aquifers was observed. This hydrogeochemical signature acted as the dominant control on denitrification reaction progress. High permeability, aerobic conditions and a lack of bacterial energy sources in the slate catchment resulted in low denitrification reaction progress (0-32%), high NO3- and comparatively low N2O emission factors (EF5g1). In the sandstone catchment denitrification progress ranged from 4 to 94% and was highly dependent on permeability, water table elevation, dissolved oxygen concentration solid phase bacterial energy sources. Denitrification of NO3- to N2 occurred in anaerobic conditions, while at intermediate dissolved oxygen; N2O was the dominant reaction product. EF5g1 (mean: 0.0018) in the denitrifying sandstone catchment was 32% less than the IPCC default. The denitrification observations across catchments were supported by stable isotope signatures. Stream NO3- occurrence was 32% lower in the sandstone catchment even though N loading was substantially higher than the slate catchment.

In situ denitrification and DNRA rates in groundwater beneath an integrated constructed wetland.

Evaluation of the environmental benefits of constructed wetlands (CWs) requires an understanding of their impacts on the groundwater quality under the wetlands. Empirical mass-balance (nitrogen in/nitrogen out) approaches for estimating nitrogen (N) removal in CWs do not characterise the final fate of N; where nitrate (NO3--N) could be reduced to either ammonium (NH4+-N) or N2 with the potential for significant production of N2O. Herein, in situ denitrification and DNRA (dissimilatory nitrate reduction to ammonium) rates were measured in groundwater beneath cells of an earthen lined integrated constructed wetland (ICW, used to remove the nutrients from municipal wastewater) using the 15N-enriched NO3--N push-pull method. Experiments were conducted utilising replicated (n = 3) shallow (1 m depth) and deep (4 m depth) piezometers installed along two control planes. These control planes allowed for the assessment of groundwater underlying high (Cell 2, septic tank waste) and low (Cell 3) load cells of the ICW. Background piezometers were also installed off-site. Results showed that denitrification (N2O-N + N2-N) and DNRA were major NO3--N consumption processes accounting together for 54-79% of the total biochemical consumption of the applied NO3--N. Of which 14-16% and 40-63% were consumed by denitrification and DNRA, respectively. Both processes differed significantly across ICW cells indicating that N transformation depends on nutrient loading rates and were significantly higher in shallow compared to the deep groundwater. In such a reduced environment (low dissolved oxygen and low redox potential), higher DNRA over the denitrification rate can be attributed to the high C concentration and high TC/NO3--N ratio. Low pH (6.5-7.1) in this system might have limited denitrification to some extent to an incomplete state, evidenced by a high N2O-N/(N2O-N+N2-N) ratio (0.35 ± 0.17, SE). A relatively higher N2O-N/(N2O-N+N2-N) ratio and higher DNRA rate over denitrification, suggest that the end products of N transformations are reactive. This N2O can be consumed to N2 and/or emitted to the atmosphere. The DNRA rate and accumulation of NH4+-N indicated that the ICW created a suitable groundwater biogeochemical environment that enhanced NO3--N reduction to NH4+-N. This study showed that CWs significantly influence NO3--N attenuation to reactive forms of N in the groundwater beneath them and that solely focusing on within wetland NO3--N attenuation can underestimate the environmental benefits of wetlands.

Improving and disaggregating N2O emission factors for ruminant excreta on temperate pasture soils.

Cattle excreta deposited on grazed grasslands are a major source of the greenhouse gas (GHG) nitrous oxide (N2O). Currently, many countries use the IPCC default emission factor (EF) of 2% to estimate excreta-derived N2O emissions. However, emissions can vary greatly depending on the type of excreta (dung or urine), soil type and timing of application. Therefore three experiments were conducted to quantify excreta-derived N2O emissions and their associated EFs, and to assess the effect of soil type, season of application and type of excreta on the magnitude of losses. Cattle dung, urine and artificial urine treatments were applied in spring, summer and autumn to three temperate grassland sites with varying soil and weather conditions. Nitrous oxide emissions were measured from the three experiments over 12months to generate annual N2O emission factors. The EFs from urine treated soil was greater (0.30-4.81% for real urine and 0.13-3.82% for synthetic urine) when compared with dung (-0.02-1.48%) treatments. Nitrous oxide emissions were driven by environmental conditions and could be predicted by rainfall and temperature before, and soil moisture deficit after application; highlighting the potential for a decision support tool to reduce N2O emissions by modifying grazing management based on these parameters. Emission factors varied seasonally with the highest EFs in autumn and were also dependent on soil type, with the lowest EFs observed from well-drained and the highest from imperfectly drained soil. The EFs averaged 0.31 and 1.18% for cattle dung and urine, respectively, both of which were considerably lower than the IPCC default value of 2%. These results support both lowering and disaggregating EFs by excreta type.

Reducing nitrous oxide emissions by changing N fertiliser use from calcium ammonium nitrate (CAN) to urea based formulations.

The accelerating use of synthetic nitrogen (N) fertilisers, to meet the world's growing food demand, is the primary driver for increased atmospheric concentrations of nitrous oxide (N2O). The IPCC default emission factor (EF) for N2O from soils is 1% of the N applied, irrespective of its form. However, N2O emissions tend to be higher from nitrate-containing fertilisers e.g. calcium ammonium nitrate (CAN) compared to urea, particularly in regions, which have mild, wet climates and high organic matter soils. Urea can be an inefficient N source due to NH3 volatilisation, but nitrogen stabilisers (urease and nitrification inhibitors) can improve its efficacy. This study evaluated the impact of switching fertiliser formulation from calcium ammonium nitrate (CAN) to urea-based products, as a potential mitigation strategy to reduce N2O emissions at six temperate grassland sites on the island of Ireland. The surface applied formulations included CAN, urea and urea with the urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT) and/or the nitrification inhibitor dicyandiamide (DCD). Results showed that N2O emissions were significantly affected by fertiliser formulation, soil type and climatic conditions. The direct N2O emission factor (EF) from CAN averaged 1.49% overall sites, but was highly variable, ranging from 0.58% to 3.81. Amending urea with NBPT, to reduce ammonia volatilisation, resulted in an average EF of 0.40% (ranging from 0.21 to 0.69%)-compared to an average EF of 0.25% for urea (ranging from 0.1 to 0.49%), with both fertilisers significantly lower and less variable than CAN. Cumulative N2O emissions from urea amended with both NBPT and DCD were not significantly different from background levels. Switching from CAN to stabilised urea formulations was found to be an effective strategy to reduce N2O emissions, particularly in wet, temperate grassland.

The effect of renovation of long-term temperate grassland on N2O emissions and N leaching from contrasting soils.

Renovation of long-term grassland is associated with a peak in soil organic N mineralisation which, coupled with diminished plant N uptake can lead to large gaseous and leaching N losses. This study reports on the effect of ploughing and subsequent N fertilisation on the N2O emissions and DON/NO3(-) leaching, and evaluates the impact of ploughing technique on the magnitude and profile of N losses. This study was carried out on isolated grassland lysimeters of three Irish soils representing contrasting drainage properties (well-drained Clonakilty, moderately-drained Elton and poorly-drained Rathangan). Lysimeters were manually ploughed simulating conventional (CT) and minimum tillage (MT) as two treatments. Renovation of grassland increased N2O flux to a maximum of 0.9kgN2O-Nha(-1) from poorly-drained soil over four days after treatment. Although there was no difference between CT and MT in the post-ploughing period, the treatment influenced subsequent N2O after fertiliser applications. Fertilisation remained the major driver of N losses therefore reducing fertilisation rate post-planting to account for N mineralised through grassland renovation could reduce the losses in medium to longer term. Leaching was a significant loss pathway, with the cumulative drainage volume and N leached highly influenced by soil type. Overall, the total N losses (N2O+N leached) were lowest from poorly and moderately draining soil and highest for the well draining soil, reflecting the dominance of leaching on total N losses and the paramount importance of soil properties.

Carbon amendment and soil depth affect the distribution and abundance of denitrifiers in agricultural soils.

The nitrite reductase (nirS and nirK) and nitrous oxide reductase-encoding (nosZ) genes of denitrifying populations present in an agricultural grassland soil were quantified using real-time polymerase chain reaction (PCR) assays. Samples from three separate pedological depths at the chosen site were investigated: horizon A (0-10 cm), horizon B (45-55 cm), and horizon C (120-130 cm). The effect of carbon addition (treatment 1, control; treatment 2, glucose-C; treatment 3, dissolved organic carbon (DOC)) on denitrifier gene abundance and N2O and N2 fluxes was determined. In general, denitrifier abundance correlated well with flux measurements; nirS was positively correlated with N2O, and nosZ was positively correlated with N2 (P < 0.03). Denitrifier gene copy concentrations per gram of soil (GCC) varied in response to carbon type amendment (P < 0.01). Denitrifier GCCs were high (ca. 10(7)) and the bac:nirK, bac:nirS, bac:nir (T) , and bac:nosZ ratios were low (ca. 10(-1)/10) in horizon A in all three respective treatments. Glucose-C amendment favored partial denitrification, resulting in higher nir abundance and higher N2O fluxes compared to the control. DOC amendment, by contrast, resulted in relatively higher nosZ abundance and N2 emissions, thus favoring complete denitrification. We also noted soil depth directly affected bacterial, archaeal, and denitrifier abundance, possibly due to changes in soil carbon availability with depth.

In situ N2O emissions are not mitigated by hippuric and benzoic acids under denitrifying conditions.

Ruminant urine patches deposited onto pasture are a significant source of greenhouse gas nitrous oxide (N2O) from livestock agriculture. Increasing food demand is predicted to lead to a rise in ruminant numbers globally, which, in turn will result in elevated levels of urine-derived N2O. Therefore mitigation strategies are urgently needed. Urine contains hippuric acid and together with one of its breakdown products, benzoic acid, has previously been linked to mitigating N2O emissions from urine patches in laboratory studies. However, the sole field study to date found no effect of hippuric and benzoic acid concentration on N2O emissions. Therefore the aim of this study was to investigate the in situ effect of these urine constituents on N2O emissions under conditions conducive to denitrification losses. Unadulterated bovine urine (0 mM of hippuric acid, U) was applied, as well as urine amended with either benzoic acid (96 mM, U+BA) or varying rates of hippuric acid (8 and 82 mM, U+HA1, U+HA2). Soil inorganic nitrogen (N) and N2O fluxes were monitored over a 66 day period. Urine application resulted in elevated N2O flux for 44 days. The largest N2O fluxes accounting for between 13% (U) and 26% (U+HA1) of total loss were observed on the day of urine application. Between 0.9 and 1.3% of urine-N was lost as N2O. Cumulative N2O loss from the control was 0.3 kg N2O-Nha(-1) compared with 11, 9, 12, and 10 kg N2O-Nha(-1) for the U, U+HA1, U+HA2, and U+BA treatments, respectively. Incremental increases in urine HA or increase in BA concentrations had no effect on N2O emissions. Although simulation of dietary manipulation to reduce N2O emissions through altering individual urine constituents appears to have no effect, there may be other manipulations such as reducing N content or inclusion of synthetic inhibitory products that warrant further investigation.

Consequences of varied soil hydraulic and meteorological complexity on unsaturated zone time lag estimates.

The true efficacy of a programme of agricultural mitigation measures within a catchment to improve water quality can be determined only after a certain hydrologic time lag period (subsequent to implementation) has elapsed. As the biophysical response to policy is not synchronous, accurate estimates of total time lag (unsaturated and saturated) become critical to manage the expectations of policy makers. The estimation of the vertical unsaturated zone component of time lag is vital as it indicates early trends (initial breakthrough), bulk (centre of mass) and total (Exit) travel times. Typically, estimation of time lag through the unsaturated zone is poor, due to the lack of site specific soil physical data, or by assuming saturated conditions. Numerical models (e.g. Hydrus 1D) enable estimates of time lag with varied levels of input data. The current study examines the consequences of varied soil hydraulic and meteorological complexity on unsaturated zone time lag estimates using simulated and actual soil profiles. Results indicated that: greater temporal resolution (from daily to hourly) of meteorological data was more critical as the saturated hydraulic conductivity of the soil decreased; high clay content soils failed to converge reflecting prevalence of lateral component as a contaminant pathway; elucidation of soil hydraulic properties was influenced by the complexity of soil physical data employed (textural menu, ROSETTA, full and partial soil water characteristic curves), which consequently affected time lag ranges; as the importance of the unsaturated zone increases with respect to total travel times the requirements for high complexity/resolution input data become greater. The methodology presented herein demonstrates that decisions made regarding input data and landscape position will have consequences for the estimated range of vertical travel times. Insufficiencies or inaccuracies regarding such input data can therefore mislead policy makers regarding the achievability of water quality targets.

Mustard catch crop enhances denitrification in shallow groundwater beneath a spring barley field.

Over-winter green cover crops have been reported to increase dissolved organic carbon (DOC) concentrations in groundwater, which can be used as an energy source for denitrifiers. This study investigates the impact of a mustard catch crop on in situ denitrification and nitrous oxide (N2O) emissions from an aquifer overlain by arable land. Denitrification rates and N2O-N/(N2O-N+N2-N) mole fractions were measured in situ with a push-pull method in shallow groundwater under a spring barley system in experimental plots with and without a mustard cover crop. The results suggest that a mustard cover crop could substantially enhance reduction of groundwater nitrate NO3--N via denitrification without significantly increasing N2O emissions. Mean total denitrification (TDN) rates below mustard cover crop and no cover crop were 7.61 and 0.002 µg kg(-1) d(-1), respectively. Estimated N2O-N/(N2O-N+N2-N) ratios, being 0.001 and 1.0 below mustard cover crop and no cover crop respectively, indicate that denitrification below mustard cover crop reduces N2O to N2, unlike the plot with no cover crop. The observed enhanced denitrification under the mustard cover crop may result from the higher groundwater DOC under mustard cover crop (1.53 mg L(-1)) than no cover crop (0.90 mg L(-1)) being added by the root exudates and root masses of mustard. This study gives insights into the missing piece in agricultural nitrogen (N) balance and groundwater derived N2O emissions under arable land and thus helps minimise the uncertainty in agricultural N and N2O-N balances.

Slow delivery of a nitrification inhibitor (dicyandiamide) to soil using a biodegradable hydrogel of chitosan.

Using chemical inhibitors to reduce soil nitrification decreases emissions of environmental damaging nitrate and nitrous oxide and improves nitrogen use efficiency in agricultural systems. The efficacy of nitrification inhibitors such as dicyandiamide (DCD) is limited in soil due to biodegradation. This study investigated if the persistence of DCD could be sustained in soil by slow release from a chitosan hydrogel. DCD was encapsulated in glyoxal-crosslinked chitosan beads where excess glyoxal was (i) partly removed (C beads) or (ii) allowed to dry (CG beads). The beads were tested in water and in soil. The beads contained two fractions of DCD: one which was quickly released in water, and one which was not. A large DCD fraction within C beads was readily available: 84% of total DCD bead content was released after 9h immersion in water, while between 74% and 98% was released after 7d in soil under low to high moisture conditions. A lower percentage of encapsulated DCD was readily released from CG beads: 19% after 9h in water, and 33% after 7d in soil under high rainfall conditions. Kinetic analysis indicated that the release in water occurred by quasi-Fickian diffusion. The results also suggest that DCD release was controlled by bead erosion and the leaching of glyoxal derivatives, predominantly a glyoxal-DCD adduct whose release was positively correlated with that of DCD (R(2)=0.99, p⩽0.0001). Therefore, novel chitosan/glyoxal composite beads show a promising slow-release potential in soil for agrochemicals like DCD.

Denitrification and indirect N2O emissions in groundwater: hydrologic and biogeochemical influences.

Identification of specific landscape areas with high and low groundwater denitrification potential is critical for improved management of agricultural nitrogen (N) export to ground and surface waters and indirect nitrous oxide (N2O) emissions. Denitrification products together with concurrent hydrogeochemical properties were analysed over two years at three depths at two low (L) and two high (H) permeability agricultural sites in Ireland. Mean N2O-N at H sites were significantly higher than L sites, and decreased with depth. Conversely, excess N2-N were significantly higher at L sites than H sites and did not vary with depth. Denitrification was a significant pathway of nitrate (NO3⁻-N) reduction at L sites but not at H sites, reducing 46-77% and 4-8% of delivered N with resulting mean NO3⁻-N concentrations of 1-4 and 12-15 mg N L⁻¹ at L and H sites, respectively. Mean N2O-N emission factors (EF5g) were higher than the most recent Intergovernmental Panel on Climate Change (IPCC, 2006) default value and more similar to the older IPCC (1997) values. Recharge during winter increased N2O but decreased excess dinitrogen (excess N2-N) at both sites, probably due to increased dissolved oxygen (DO) coupled with low groundwater temperatures. Denitrifier functional genes were similar at all sites and depths. Data showed that highly favourable conditions prevailed for denitrification to occur--multiple electron donors, low redox potential (Eh<100 mV), low DO (<2 mg L⁻¹), low permeability (k(s)<0.005 m·d⁻¹) and a shallow unsaturated zone (<2 m). Quantification of excess N2-N in groundwater helps to close N balances at the local, regional and global scales.

Evaluation of headspace equilibration methods for quantifying greenhouse gases in groundwater.

The objective of the study was to evaluate the different headspace equilibration methods for the quantification of dissolved greenhouse gases in groundwater. Groundwater samples were collected from wells with contrasting hydrogeochemical properties and degassed using the headspace equilibration method. One hundred samples from each well were randomly selected, treatments were applied and headspace gases analysed by gas chromatography. Headspace equilibration treatments varied helium (He):water ratio, shaking time and standing time. Mean groundwater N(2)O, CO(2) and CH(4) concentrations were 0.024 mg N L(-1), 13.71 mg C L(-1) and 1.63 µg C L(-1), respectively. All treatments were found to significantly influence dissolved gas concentrations. Considerable differences in the optimal He:water ratio and standing time were observed between the three gases. For N(2)O, CO(2) and CH(4) the optimum operating points for He:water ratio was 4.4:1, 3:1 and 3.4:1; shaking time was 13, 12 and 13 min; and standing time was 63, 17 and 108 min, respectively. The headspace equilibration method needs to be harmonised to ensure comparability between studies. The experiment reveals that He:water ratio 3:1 and shaking time 13 min give better estimation of dissolved gases than any lower or higher ratios and shaking times. The standing time 63, 17 and 108 min should be applied for N(2)O, CO(2) and CH(4), respectively.