Geochemical and isotopic study of abiotic nitrite reduction coupled to biologically produced Fe(II) oxidation in marine environments.

Estuarine sediments are often characterized by abundant iron oxides, organic matter, and anthropogenic nitrogen compounds (e.g., nitrate and nitrite). Anoxic dissimilatory iron reducing bacteria (e.g., Shewanella loihica) are ubiquitous in these environments where they can catalyze the reduction of Fe(III) (oxyhydr)oxides, thereby releasing aqueous Fe(II). The biologically produced Fe(II) can later reduce nitrite to form nitrous oxide. The effect on nitrite reduction by both biologically produced and artificially amended Fe(II) was examined experimentally. Ferrihydrite was reduced by Shewanella loihica in a batch reaction with an anoxic synthetic sea water medium. Some of the Fe(II) released by S. loihica adsorbed onto ferrihydrite, which was involved in the transformation of ferrihydrite to magnetite. In a second set of experiments with identical medium, no microorganism was present, instead, Fe(II) was amended. The amount of solid-bound Fe(II) in the experiments with bioproduced Fe(II) increased the rate of abiotic NO2- reduction with respect to that with synthetic Fe(II), yielding half-lives of 0.07 and 0.47 d, respectively. The δ18O and δ15N of NO2- was measured through time for both the abiotic and innoculated experiments. The ratio of ε18O/ε15N was 0.6 for the abiotic experiments and 3.1 when NO2- was reduced by S. loihica, thus indicating two different mechanisms for the NO2- reduction. Notably, there is a wide range of the ε18O/ε15N values in the literature for abiotic and biotic NO2- reduction, as such, the use of this ratio to distinguish between reduction mechanisms in natural systems should be taken with caution. Therefore, we suggest an additional constraint to identify the mechanisms (i.e. abiotic/biotic) controlling NO2- reduction in natural settings through the correlation of δ15N-NO2- and the aqueous Fe(II) concentration.

A multi-isotopic approach to investigate the influence of land use on nitrate removal in a highly saline lake-aquifer system.

Endorheic or closed drainage basins in arid and semi-arid regions are vulnerable to pollution. Nonetheless, in the freshwater-saltwater interface of endorheic saline lakes, oxidation-reduction (redox) reactions can attenuate pollutants such as nitrate (NO3-). This study traces the ways of nitrogen (N) removal in the Pétrola lake-aquifer system (central Spain), an endorheic basin contaminated with NO3- (up to 99.2mg/L in groundwater). This basin was declared vulnerable to NO3- pollution in 1998 due to the high anthropogenic pressures (mainly agriculture and wastewaters). Hydrochemical, multi-isotopic (δ18ONO3, δ15NNO3, δ13CDIC, δ18OH2O, and δ2HH2O) and geophysical techniques (electrical resistivity tomography) were applied to identify the main redox processes at the freshwater-saltwater interface. The results showed that the geometry of this interface is influenced by land use, causing spatial variability of nitrogen biogeochemical processes over the basin. In the underlying aquifer, NO3- showed an average concentration of 38.5mg/L (n=73) and was mainly derived from agricultural inputs. Natural attenuation of NO3- was observed in dryland farming areas (up to 72%) and in irrigation areas (up to 66%). In the Pétrola Lake, mineralization and organic matter degradation in lake sediment play an important role in NO3- reduction. Our findings are a major step forward in understanding freshwater-saltwater interfaces as reactive zones for NO3- attenuation. We further emphasize the importance of including a land use perspective when studying water quality-environmental relationships in hydrogeological systems dominated by density-driven circulation.

Tracing the role of endogenous carbon in denitrification using wine industry by-product as an external electron donor: Coupling isotopic tools with mathematical modeling.

Nitrate removal through enhanced biological denitrification (EBD), consisting of the inoculation of an external electron donor, is a feasible solution for the recovery of groundwater quality. In this context, liquid waste from wine industries (wine industry by-products, WIB) may be feasible for use as a reactant to enhance heterotrophic denitrification. To address the feasibility of WIB as electron donor to promote denitrification, as well as to evaluate the role of biomass as a secondary organic C source, a flow-through experiment was carried out. Chemical and isotopic characterization was performed and coupled with mathematical modeling. Complete nitrate attenuation with no nitrite accumulation was successfully achieved after 10 days. Four different C/N molar ratios (7.0, 2.0, 1.0 and 0) were tested. Progressive decrease of the C/N ratio reduced the remaining C in the outflow and favored biomass migration, producing significant changes in dispersivity in the reactor, which favored efficient nitrate degradation. The applied mathematical model described the general trends for nitrate, ethanol, dissolved organic carbon (DOC) and dissolved inorganic carbon (DIC) concentrations. This model shows how the biomass present in the system is degraded to dissolved organic C (DOCen) and becomes the main source of DOC for a C/N ratio between 1.0 and 0. The isotopic model developed for organic and inorganic carbon also describes the general trends of δ13C of ethanol, DOC and DIC in the outflow water. The study of the evolution of the isotopic fractionation of organic C using a Rayleigh distillation model shows the shift in the organic carbon source from the WIB to the biomass and is in agreement with the isotopic fractionation values used to calibrate the model. Isotopic fractionations (ε) of C-ethanol and C-DOCen were -1‰ and -5‰ (model) and -3.3‰ and -4.8‰ (Rayleigh), respectively. In addition, an inverse isotopic fractionation of +10‰ was observed for biomass degradation to DOCen. Overall, WIB can efficiently promote nitrate reduction in EBD treatments. The conceptual model of the organic C cycle and the developed mathematical model accurately described the chemical and isotopic transformations that occur during this induced denitrification.

Denitrification in a hypersaline lake-aquifer system (Pétrola Basin, Central Spain): the role of recent organic matter and Cretaceous organic rich sediments.

Agricultural regions in semi-arid to arid climates with associated saline wetlands are one of the most vulnerable environments to nitrate pollution. The Pétrola Basin was declared vulnerable to NO3(-) pollution by the Regional Government in 1998, and the hypersaline lake was classified as a heavily modified body of water. The study assessed groundwater NO3(-) through the use of multi-isotopic tracers (δ(15)N, δ(34)S, δ(13)C, δ(18)O) coupled to hydrochemistry in the aquifer connected to the eutrophic lake. Hydrogeologically, the basin shows two main flow components: regional groundwater flow from recharge areas (Zone 1) to the lake (Zone 2), and a density-driven flow from surface water to the underlying aquifer (Zone 3). In Zones 1 and 2, δ(15)NNO3 and δ(18)ONO3 suggest that NO3(-) from slightly volatilized ammonium synthetic fertilizers is only partially denitrified. The natural attenuation of NO3(-) can occur by heterotrophic reactions. However, autotrophic reactions cannot be ruled out. In Zone 3, the freshwater-saltwater interface (down to 12-16 m below the ground surface) is a reactive zone for NO3(-) attenuation. Tritium data suggest that the absence of NO3(-) in the deepest zones of the aquifer under the lake can be attributed to a regional groundwater flow with long residence time. In hypersaline lakes the geometry of the density-driven flow can play an important role in the transport of chemical species that can be related to denitrification processes.

Feeding strategies for groundwater enhanced biodenitrification in an alluvial aquifer: chemical, microbial and isotope assessment of a 1D flow-through experiment.

Nitrate-removal through enhanced in situ biodenitrification (EISB) is an existing alternative for the recovery of groundwater quality, and is often suggested for use in exploitation wells pumping at small flow-rates. Innovative approaches focus on wider-scale applications, coupling EISB with water-management practices and new monitoring tools. However, before this approach can be used, some water-quality issues such as the accumulation of denitrification intermediates and/or of reduced compounds from other anaerobic processes must be addressed. With such a goal, a flow-through experiment using 100mg-nitrate/L groundwater was built to simulate an EISB for an alluvial aquifer. Heterotrophic denitrification was induced through the periodic addition of a C source (ethanol), with four different C addition strategies being evaluated to improve the quality of the denitrified water. Chemical, microbial and isotope analyses of the water were performed. Biodenitrification was successfully stimulated by the daily addition of ethanol, easily achieving drinking water standards for both nitrate and nitrite, and showing an expected linear trend for nitrogen and oxygen isotope fractionation, with a εN/εO value of 1.1. Nitrate reduction to ammonium was never detected. Water quality in terms of remaining C, microbial counts, and denitrification intermediates was found to vary with the experimental time, and some secondary microbial respiration processes, mainly manganese reduction, were suspected to occur. Carbon isotope composition from the remaining ethanol also changed, from an initial enrichment in (13)C-ethanol compared to the value of the injected ethanol (-30.6‰), to a later depletion, achieving δ(13)C values well below the initial isotope composition (to a minimum of -46.7‰). This depletion in the heavy C isotope follows the trend of an inverse fractionation. Overall, our results indicated that most undesired effects on water quality may be controlled through the optimization of the C/N ratio determined from the amounts of injected ethanol vs. the amount of nitrate in groundwater, with a smaller C/N ratio causing a lower level of undesired impurities. Furthermore, the authors suggest that the biofilm life-time has a direct effect on microbial population and hence affects biodenitrification performance, influencing the accumulation of nitrite over time.

Nitrate attenuation potential of hypersaline lake sediments in central Spain: flow-through and batch experiments.

Complex lacustrine systems, such as hypersaline lakes located in endorheic basins, are exposed to nitrate (NO3(-)) pollution. An excellent example of these lakes is the hypersaline lake located in the Pétrola basin (central Spain), where the lake acts as a sink for NO3(-) from agricultural activities and from sewage from the surrounding area. To better understand the role of the organic carbon (Corg) deposited in the bottom sediment in promoting denitrification, a four-stage flow-through experiment (FTR) and batch experiments using lake bottom sediment were performed. The chemical, multi-isotopic and kinetic characterization of the outflow showed that the intrinsic NO3(-) attenuation potential of the lake bottom sediment was able to remove 95% of the NO3(-) input over 296days under different flow conditions. The NO3(-) attenuation was mainly linked with denitrification but some dissimilatory nitrate reduction to ammonium was observed at early days favored by the high C/N ratio and salinity. Sulfate reduction could be neither confirmed nor discarded during the experiments because the sediment leaching masked the chemical and isotopic signatures of this reaction. The average nitrogen reduction rate (NRR) obtained was 1.25mmold(-1)kg(-1) and was independent of the flow rate employed. The amount of reactive Corg from the bottom sediment consumed during denitrification was 28.8mmol, representing approximately 10% of the total Corg of the sediment (1.2%). Denitrification was produced coupled with an increase in the isotopic composition of both δ(15)N and δ(18)O. The isotopic fractionations (ε of (15)N-NO3(-) and (18)O-NO3(-)) produced during denitrification were calculated using batch and vertical profile samples. The results were -14.7‰ for εN and -14.5‰ for εO.

Changes of hormones regulating electrolyte metabolism after space flight and hypokinesia.

The changes of hormones in plasma involved in the body fluid regulation were studied in human subjects during and after space flights in relation to redistribution of body fluids in the state of weightlessness. Since hypokinesia was used as a model for simulation of some effects of the stay in microgravity the plasma hormone levels in rats exposed to hypokinesia were also investigated. Plasma aldosterone values showed great individual variations during the first inflight days, the increased levels were observed with prolongation of space flights. The important elevation was found in the recovery period, however it was interesting to note, that in some cosmonauts with repeated exposure to space flight, the postflight plasma aldosterone levels were not elevated. The urine excretion of aldosterone was increased inflight, however in postflight period the decrease or increase were found in the first 1-5 days. The increase of plasma renin activity was observed in flight and postflight period. The rats were exposed to hypokinesia (forced restriction of motor activity) for 1, 7 and 60 days and urine was collected during last 24 hours. The animals were sacrificed and the concentration of electrolytes and of levels of corticosterone, aldosterone (A), ANF and plasma-renin activity (PRA) were determined in plasma. In urine excretion of sodium and potassium were estimated. An important increase of plasma renin activity and aldosterone concentration was found after short-term hypokinesia (1 day). These hormonal values appear to decrease with time (7 days) and are not significantly different from controls after long-term hypokinesia (60 days). A decrease of values ANF in plasma was observed after 1 and 7 days hypokinesia. After prolonged hypokinesia a decrease of sodium plasma concentration was observed. The excretion of sodium in urine was higher in long-term hypokinetic animals. There were no significant changes of plasma potassium levels in rats exposed to hypokinesia, however the urinary excretion of potassium was elevated. In rats exposed to hypokinesia for 7 and 60 days an increase of urine osmolality was observed. The results of hormone and electrolyte determination in plasma of cosmonauts after space flight and in experimental animals after hypokinesia suggested that in evaluation of relations between the changes of hormone levels and electrolyte in plasma and urine other factors like emotional stress working load; altered diurnal cycles should be considered in interpretation of homeostatic response of fluid and electrolyte metabolism to space flight conditions.