A novel 15N tracer approach for the quantification of N2 and N2O emissions from soil incubations in a completely automated laboratory set up

The microbial mediated production of nitrous oxide (N2O) and its reduction to dinitrogen (N2) via denitrification represents a loss of nitrogen (N) from fertilised agro-ecosystems to the atmosphere. Although denitrification has received great interest by biogeochemists in the last decades, the magnitude of N2lossesand related N2:N2O ratios from soils still are largely unknown due to methodical constraints. We present a novel 15N tracer approach, based on a previous developed tracer method to study denitrification in pure bacterial cultures which was modified for the use on soil incubations in a completely automated laboratory set up. The method uses a background air in the incubation vessels that is replaced with a helium-oxygen gas mixture with a 50-fold reduced N2 background (2 % v/v). This method allows for a direct and sensitive quantification of the N2 and N2O emissions from the soil with isotope-ratio mass spectrometry after 15N labelling of denitrification N substrates and minimises the sensitivity to the intrusion of atmospheric N2 at the same time. The incubation set up was used to determine the influence of different soil moisture levels on N2 and N2O emissions from a sub-tropical pasture soil in Queensland/Australia. The soil was labelled with an equivalent of 50 μg-N per gram dry soil by broadcast application of KNO3solution (4 at.% 15N) and incubated for 3 days at 80% and 100% water filled pore space (WFPS), respectively. The headspace of the incubation vessel was sampled automatically over 12hrs each day and 3 samples (0, 6, and 12 hrs after incubation start) of headspace gas analysed for N2 and N2O with an isotope-ratio mass spectrometer (DELTA V Plus, Thermo Fisher Scientific, Bremen, Germany(. In addition, the soil was analysed for 15N NO3- and NH4+ using the 15N diffusion method, which enabled us to obtain a complete N balance. The method proved to be highly sensitive for N2 and N2O emissions detecting N2O emissions ranging from 20 to 627 μN kg-1soil-1hr-1and N2 emissions ranging from 4.2 to 43 μN kg-1soil-1hr-1for the different treatments. The main end-product of denitrification was N2O for both water contents with N2 accounting for 9% and 13% of the total denitrification losses at 80% and 100%WFPS, respectively. Between 95-100% of the added 15N fertiliser could be recovered. Gross nitrification over the 3 days amounted to 8.6 μN g-1 soil-1 and 4.7 μN g-1 soil-1, denitrification to 4.1 μN g-1 soil-1 and 11.8 μN g-1 soil-1at 80% and 100%WFPS, respectively. The results confirm that the tested method allows for a direct and highly sensitive detection of N2 and N2O fluxes from soils and hence offers a sensitive tool to study denitrification and N turnover in terrestrial agro-ecosystems.

Behavioral and EEG effects of GABAergic manipulation of the nigro-tectal pathway in the Wistar audiogenic rat (WAR) strain II: An EEG wavelet analysis and retrograde neuronal tracer approach

The role of the substantia nigra pars reticulata (SNPr) and superior colliculus (SC) network in rat strains susceptible to audiogenic seizures still remain underexplored in epileptology. In a previous study from our laboratory, the GABAergic drugs bicuculline (BIC) and muscimol (MUS) were microinjected into the deep layers of either the anterior SC (aSC) or the posterior SC (pSC) in animals of the Wistar audiogenic rat (WAR) strain submitted to acoustic stimulation, in which simultaneous electroencephalographic (EEG) recording of the aSC, pSC, SNPr and striatum was performed. Only MUS microinjected into the pSC blocked audiogenic seizures. In the present study, we expanded upon these previous results using the retrograde tracer Fluorogold (FG) microinjected into the aSC and pSC in conjunction with quantitative EEG analysis (wavelet transform), in the search for mechanisms associated with the susceptibility of this inbred strain to acoustic stimulation. Our hypothesis was that the WAR strain would have different connectivity between specific subareas of the superior colliculus and the SNPr when compared with resistant Wistar animals and that these connections would lead to altered behavior of this network during audiogenic seizures. Wavelet analysis showed that the only treatment with an anticonvulsant effect was MUS microinjected into the pSC region, and this treatment induced a sustained oscillation in the theta band only in the SNPr and in the pSC. These data suggest that in WAR animals, there are at least two subcortical loops and that the one involved in audiogenic seizure susceptibility appears to be the pSC-SNPr circuit. We also found that WARs presented an increase in the number of FG + projections from the posterior SNPr to both the aSC and pSC (primarily to the pSC), with both acting as proconvulsant nuclei when compared with Wistar rats. We concluded that these two different subcortical loops within the basal ganglia are probably a consequence of the WAR genetic background. (C) 2012 Elsevier Inc. All rights reserved.

Quantifying nitrogen cycling rates in freshwater marshes of the southern Everglades using nitrogen-15 tracer techniques

This dissertation research project addressed the question of how hydrologic restoration of the Everglades is impacting the nutrient dynamics of marsh ecosystems in the southern Everglades. These effects were analyzed by quantifying nitrogen (N) cycle dynamics in the region. I utilized stable isotope tracer techniques to investigate nitrogen uptake and cycling between the major ecosystem components of the freshwater marsh system. I recorded the natural isotopic signatures (δ15N and δ 13C) for major ecosystem components from the three major watersheds of the Everglades: Shark River Slough, Taylor Slough, and C-111 basin. Analysis of δ15 N and δ13C natural abundance data were used to demonstrate the spatial extent to which nitrogen from anthropogenic or naturally enriched sources is entering the marshes of the Everglades. In addition, I measured the fluxes on N between various ecosystem components at both near-canal and estuarine ecotone locations. Lastly, I investigated the effect of three phosphorus load treatments (0.00 mg P m-2, 6.66 mg P m-2, and 66.6 mg P m-2) on the rate and magnitude of ecosystem N-uptake and N-cycling. The δ15N and δ13C natural abundance data supported the hypothesis that ecosystem components from near-canal sites have heavier, more enriched δ 15N isotopic signatures than downstream sites. The natural abundance data also showed that the marshes of the southern Everglades are acting as a sink for isotopically heavier, canal-borne dissolved inorganic nitrogen (DIN) and a source for "new" marsh derived dissolved organic nitrogen (DON). In addition, the 15N mesocosm data showed the rapid assimilation of the 15N tracer by the periphyton component and the delayed N uptake by soil and macrophyte components in the southern Everglades.

Tracing of 15N in a mountain spruce forest: comparison of experimental data with th TRACE model

Despite numerous studies about nitrogen-cycling in forest ecosystems, many uncertainties remain, especially regarding the longer-term nitrogen accumulation. To contribute to filling this gap, the dynamic process-based model TRACE, with the ability to simulate 15N tracer redistribution in forest ecosystems was used to study N cycling processes in a mountain spruce forest of the northern edge of the Alps in Switzerland (Alptal, SZ). Most modeling analyses of N-cycling and C-N interactions have very limited ability to determine whether the process interactions are captured correctly. Because the interactions in such a system are complex, it is possible to get the whole-system C and N cycling right in a model without really knowing if the way the model combines fine-scale interactions to derive whole-system cycling is correct. With the possibility to simulate 15N tracer redistribution in ecosystem compartments, TRACE features a very powerful tool for the validation of fine-scale processes captured by the model. We first adapted the model to the new site (Alptal, Switzerland; long-term low-dose N-amendment experiment) by including a new algorithm for preferential water flow and by parameterizing of differences in drivers such as climate, N deposition and initial site conditions. After the calibration of key rates such as NPP and SOM turnover, we simulated patterns of 15N redistribution to compare against 15N field observations from a large-scale labeling experiment. The comparison of 15N field data with the modeled redistribution of the tracer in the soil horizons and vegetation compartments shows that the majority of fine-scale processes are captured satisfactorily. Particularly, the model is able to reproduce the fact that the largest part of the N deposition is immobilized in the soil. The discrepancies of 15N recovery in the LF and M soil horizon can be explained by the application method of the tracer and by the retention of the applied tracer by the well developed moss layer, which is not considered in the model. Discrepancies in the dynamics of foliage and litterfall 15N recovery were also observed and are related to the longevity of the needles in our mountain forest. As a next step, we will use the final Alptal version of the model to calculate the effects of climate change (temperature, CO2) and N deposition on ecosystem C sequestration in this regionally representative Norway spruce (Picea abies) stand.

Assessing agronomic and environmental implications of different N fertilisation strategies in subtropical grain cropping systems on Oxisols

A multi-season 15N tracer recovery experiment was conducted on an Oxisol cropped with wheat, maize and sorghum to compare crop N recoveries of different fertilisation strategies and determine the main pathways of N losses that limit N recovery in these agroecosystems. In the wheat and maize seasons, 15N-labelled fertiliser was applied as conventional urea (CONV) and urea coated with a nitrification inhibitor (DMPP). In sorghum, the fate of 15N-labelled urea was monitored in this crop following a legume ley pasture (L70) or a grass ley pasture (G100). The fertiliser N applied to sorghum in the legume-cereal rotation was reduced (70 kg N ha−1) compared to the grass-cereal (100 kg N ha−1) to assess the availability of the N residual from the legume ley pasture. Average crop N recoveries were 73 % (CONV) and 77 % (DMPP) in wheat and 50 % (CONV) and 51 % (DMPP) in maize, while in sorghum were 71 % (L70) and 53 % (G100). Data gathered in this study indicate that the intrinsic physical and chemical conditions of Oxisols can be extremely effective in limiting N losses via deep leaching or denitrification. Elevated crop 15N recoveries can be therefore obtained in subtropical Oxisols using conventional urea while in these agroecosystems DMPP urea has no significant scope to increase fertiliser N recovery in the crop. Overall, introducing a legume phase to limit the fertiliser N requirements of the following cereal crop proved to be the most effective strategy to reduce N losses and increase fertiliser N recovery.

Bedeutung der C- und N-Rhizodeposition für das mikrobielle Wachstum und den mikrobiellen Umsatz von C und N in der Rhizosphäre in Abhängigkeit von der Entfernung zur Wurzel

Ziel dieser Arbeit war es, die Bedeutung des C und N mit Herkunft aus der Rhizodeposition für das mikrobielle Wachstum und den C- und N-Umsatz in der Rhizosphäre in Abhängigkeit von der Entfernung zur Wurzel zu untersuchen. Dazu wurde als wesentliche methodische Voraussetzung ein künstliches Rhizosphärensystem entwickelt, um die durch die Rhizodeposite induzierten Prozesse an der Grenzfläche zwischen Wurzeloberfläche und Boden zu untersuchen. Dieses eingesetzte Rhizosphärensystem wurde nach einem Vorbild eines in schon vielfältigen Untersuchungen der Rhizosphäre eingesetzten Systems von GAHOONIA und NIELSEN (1991) konzipiert. Zur Verfolgung der pflanzlichen C- und N-Rhizodeposition im Boden wurden 13C- und 15N-Tracer-Isotopen-Techniken zur Isotopenmarkierung der Testpflanzen eingesetzt. Zur Probenahme des Rhizosphärenbodens in räumlichen Abstand zur künstlichen Rhizoplane wurde eine Schneidvorrichtung nach FITZ et al. (2003) eingesetzt, die es ermöglicht frische Bodenproben in definiertem Abstand zur künstlichen Rhizoplane zu schneiden. Das Rhizosphärensystem wurde in 13C- und 15N-Doppelmarkierungsexperimenten mit Lolium perenne, Triticum aestivum und Avena sativa eingesetzt.Das unterschiedliche Ansprechen der mikrobiellen Gemeinschaft auf den Substrateintrag in unterschiedlicher Entfernung zur Wurzel zeigte komplexe Wechselwirkungen in Bezug auf das mikrobielle Wachstum, den mikrobiellen Umsatz, den Substrateintrag aus der Rhizodeposition und den stimulierten Abbau der nativen organischen Bodensubstanz. Der Eintrag von Rhizodepositen stellt daher eine bedeutende Funktion in der Regulation der mikrobiellen Biomasse und der Prozesse des Umsatzes der organischen Bodensubstanz während und wahrscheinlich bis nach der Vegetationsperiode dar. Die simultane Verfolgung von Rhizodepositions-C und -N im Boden und deren Funktionen innerhalb der Rhizosphäre, gerade im Hinblick auf die mikrobielle Biomasse und den Umsatz der organischen Substanz im Boden, führt zu einem besseren Verständnis der komplexen wechselseitigen Prozesse zwischen Pflanze und Boden.

Investigation of young water inflow in karst aquifers using SF6–CFC–3H/He–85Kr–39Ar and stable isotope components

Karst aquifers are known for their wide distribution of water transfer velocities. From this observation, a multiple geochemical tracer approach seems to be particularly well suited to provide a significant assessment of groundwater flows, but the choice of adapted tracers is essential. In this study, several common tracers in karst aquifers such as physicochemical parameters, major ions, stable isotopes, and d13C to more specific tracers such as dating tracers – 14C, 3H, 3H–3He, CFC-12, SF6 and 85Kr, and 39Ar – were used, in a fractured karstic carbonated aquifer located in Burgundy (France). The information carried by each tracer and the best sampling strategy are compared on the basis of geochemical monitoring done during several recharge events and over longer time periods (months to years). This study’s results demonstrate that at the seasonal and recharge event time scale, the variability of concentrations is low for most tracers due to the broad spectrum of groundwater mixings. The tracers used traditionally for the study of karst aquifers, i.e., physicochemical parameters and major ions, efficiently describe hydrological processes such as the direct and differed recharge, but require being monitored at short time steps during recharge events to be maximized. From stable isotopes, tritium, and Cl� contents, the proportion of the fast direct recharge by the largest porosity was estimated using a binary mixing model. The use of tracers such as CFC-12, SF6, and 85Kr in karst aquifers provides additional information, notably an estimation of apparent age, but they require good preliminary knowledge of the karst system to interpret the results suitably. The CFC-12 and SF6 methods efficiently determine the apparent age of baseflow, but it is preferable to sample the groundwater during the recharge event. Furthermore, these methods are based on different assumptions such as regional enrichment in atmospheric SF6, excess air, and flow models among others. 85Kr and 39Ar concentrations can potentially provide a more direct estimation of groundwater residence time. Conversely, the 3H–3He method is inefficient in the karst aquifer for dating due to 3He degassing.

Geochemical studies of surface sediment samples off North-West Africa

One hundred and twenty-eight surface-sediment samples collected off North-West Africa were studied geochemically to detect the expressions of different meridional climate regimes and zonal productivity gradients in the surface sediments. This geochemical multi-tracer approach, coupled with additional information on the bulk carbonate and TOC contents makes it possible to characterise the sedimentological regime in detail. Typical terrigenous elements like Al, K and Fe mirror the importance of the humid (fluvial) influence in the north of the study area and the dominance of aeolian input in the south. Furthermore, the distributions of Ti and Fe in the surface sediments serve as tracers for the supply of eolian volcanic material from the Canary Islands. The spatial variability of the TOC contents in the surface sediments closely follows the ocean surface productivity patterns, whereas the CaCO3 contents are mainly controlled by dilution with terrigenous matter. The potential productivity proxy Ba is not a reliable tracer for productivity in this region, since it is mainly supplied by terrigenous input (coupled with aluminosilicates).

Identification of the causes of subsoil ammonium accumulations in southeastern Queensland

Unusually high concentrations of exchangeable-NH4+ (up to 270 kg-N/ha) were observed in a Vertisol below 1 m in southeast Queensland. This study aimed to identify the source of this NH4+. Preliminary sampling of native vegetation and cropping areas had found that elevated NH4+was only present under cropped soil, indicating that clearing was linked to the NH4+formation. Mechanisms of NH4+formation that may have occurred in the subsoil after clearing were hypothesised to be a) mineralisation of organic-N; b) NO3- reduction to NH4+; and/or c) the release of fixed-NH4+. In addition it was proposed that nitrification was inhibited in the subsoil, and that this allowed any NH4+formed to accumulate over time. Incubation experiments to examine nitrification rates revealed that nitrification was undetectable, and appeared to be limited by a combination of subsoil acidity and low numbers of nitrifying organisms. Mineralisation studies also revealed that the mineralisation of organic-N was undetectable, and that mineralising organisms were limited by acidity. A small amount of nitrate ammonification could be observed with the aid of a 15N tracer if the soil was waterlogged. However, this NH4+was insufficient to account for the overall NH4+accumulation, and these waterlogged conditions were not observed in the field. Concentrations of fixed- NH4+ measured were also too low to have been responsible for the accumulation of exchangeable-NH4+. It was concluded that none of the proposed hypotheses of NH4+formation could account for the NH4+accumulation observed.

Composition of sediments from the Arabian Sea

Thirty seven deep-sea sediment cores from the Arabian Sea were studied geochemically (49 major and trace elements) for four time slices during the Holocene and the last glacial, and in one high sedimentation rate core (century scale resolution) to detect tracers of past variations in the intensity of the atmospheric monsoon circulation and its hydrographic expression in the ocean surface. This geochemical multi-tracer approach, coupled with additional information on the grain size composition of the clastic fraction, the bulk carbonate and biogenic opal contents makes it possible to characterize the sedimentological regime in detail. Sediments characterized by a specific elemental composition (enrichment) originated from the following sources: river suspensions from the Tapti and Narbada, draining the Indian Deccan traps (Ti, Sr); Indus sediments and dust from Rajasthan and Pakistan (Rb, Cs); dust from Iran and the Persian Gulf (Al, Cr); dust from central Arabia (Mg); dust from East Africa and the Red Sea (Zr/Hf, Ti/Al). Corg, Cd, Zn, Ba, Pb, U, and the HREE are associated with the intensity of upwelling in the western Arabian Sea, but only those patterns that are consistently reproduced by all of these elements can be directly linked with the intensity of the southwest monsoon. Relying on information from a single element can be misleading, as each element is affected by various other processes than upwelling intensity and nutrient content of surface water alone. The application of the geochemical multi-tracer approach indicates that the intensity of the southwest monsoon was low during the LGM, declined to a minimum from 15,000-13,000 14C year BP, intensified slightly at the end of this interval, was almost stable during the Bölling, Alleröd and the Younger Dryas, but then intensified in two abrupt successions at the end of the Younger Dryas (9900 14C year BP) and especially in a second event during the early Holocene (8800 14C year BP). Dust discharge by northwesterly winds from Arabia exhibited a similar evolution, but followed an opposite course: high during the LGM with two primary sources-the central Arabian desert and the dry Persian Gulf region. Dust discharge from both regions reached a pronounced maximum at 15,000-13,000 14C year. At the end of this interval, however, the dust plumes from the Persian Gulf area ceased dramatically, whereas dust discharge from central Arabia decreased only slightly. Dust discharge from East Africa and the Red Sea increased synchronously with the two major events of southwest monsoon intensification as recorded in the nutrient content of surface waters. In addition to the tracers of past dust flux and surface water nutrient content, the geochemical multi-tracer approach provides information on the history of deep sea ventilation (Mo, S), which was much lower during the last glacial maximum than during the Holocene. The multi-tracer approach-i.e. a few sedimentological parameters plus a set of geochemical tracers widely available from various multi-element analysis techniques-is a highly applicable technique for studying the complex sedimentation patterns of an ocean basin, and, specifically in the case of the Arabian Sea, can even reveal the seasonal structure of climate change.

Dispersivity Determination Through a Modeling Approach From a Tracer Test Based on Total Br Concentration in Soil Samples

Determination of reliable solute transport parameters is an essential aspect for the characterization of the mechanisms and processes involved in solute transport (e.g., pesticides, fertilizers, contaminants) through the unsaturated zone. A rapid inexpensive method to estimate the dispersivity parameter at the field scale is presented herein. It is based on the quantification by the X-ray fluorescence solid-state technique of total bromine in soil, along with an inverse numerical modeling approach. The results show that this methodology is a good alternative to the classic Br− determination in soil water by ion chromatography. A good agreement between the observed and simulated total soil Br is reported. The results highlight the potential applicability of both combined techniques to infer readily solute transport parameters under field conditions.