Peter Bondo Christensen

Electric currents couple spatially separated biogeochemical processes in marine sediment

Some bacteria are capable of extracellular electron transfer, thereby enabling them to use electron acceptors and donors without direct cell contact. Beyond the micrometre scale, however, no firm evidence has previously existed that spatially segregated biogeochemical processes can be coupled by electric currents in nature. Here we provide evidence that electric currents running through defaunated sediment couple oxygen consumption at the sediment surface to oxidation of hydrogen sulphide and organic carbon deep within the sediment. Altering the oxygen concentration in the sea water overlying the sediment resulted in a rapid (<1-h) change in the hydrogen sulphide concentration within the sediment more than 12 mm below the oxic zone, a change explicable by transmission of electrons but not by diffusion of molecules. Mass balances indicated that more than 40% of total oxygen consumption in the sediment was driven by electrons conducted from the anoxic zone. A distinct pH peak in the oxic zone could be explained by electrochemical oxygen reduction, but not by any conventional sets of aerobic sediment processes. We suggest that the electric current was conducted by bacterial nanowires combined with pyrite, soluble electron shuttles and outer-membrane cytochromes. Electrical communication between distant chemical and biological processes in nature adds a new dimension to our understanding of biogeochemistry and microbial ecology.

Effects of salinity on NH4+ adsorption capacity, nitrification, and denitrification in Danish estuarine sediments

The regulatory effect of salinity on nitrogen dynamics in estuarine sediments was investigated in the Randers Fjord estuary, Denmark, using sediment slurries and intact sediment cores and applying 15N-isotope techniques. Sediment was sampled at three representative stations varying in salinity, and all experiments were run at 0‰, 10‰, 20‰, and 30‰. The sediment NH4+ adsorption capacity decreased markedly at all stations when salinity was increased from 0‰ to 10‰; further increase showed little effect. In situ nitrification and denitrification also decreased with increasing salinities, with the most pronounced reduction of approximately 50% being observed when the salinity was raised from 0‰ to 10‰. The salinity-induced reduction in NH4+ adsorption capacity and stimulation of NH4+ efflux has previously been argued to cause a reduction in nitrification activity since the nitrifying bacteria become limited by NH4+ availability at higher salinities. However, using a potential nitrification assay where NH4+ was added in excess, it was demonstrated that potential nitrification activity also decreased with increasing salinity, indicating that the inhibitory salinity effect may also be a physiological effect on the microorganisms. This hypothesis was supported by the finding that denitrification based on NO3− from the overlying water (Dw), which is independent of the nitrification process, and hence NH4+ availability, also decreased with increasing salinity. We conclude that changes in salinity have a significant effect on nitrogen dynamics in estuarine sediments, which must be considered when nitrogen transformations are measured and evaluated.

LONG‐TERM CHANGES AND IMPACTS OF HYPOXIA IN DANISH COASTAL WATERS

A 38-year record of bottom water dissolved oxygen concentrations in coastal marine ecosystems around Denmark (1965-2003) and a longer partially reconstructed record of total nitrogen (TN) inputs (1900-2003) were assembled to describe long-term patterns in hypoxia and anoxia. Interannual variations in bottom water oxygen concentrations were analyzed in relation to various explanatory variables (bottom temperature, wind speed, advective transport, TN loading). Reconstructed TN loads peaked in the 1980s with a gradual decline to the present, commensurate with a legislated nutrient reduction strategy. Mean bottom water oxygen concentrations during summer have significantly declined in coastal marine ecosystems, decreasing substantially during the 1980s and were extremely variable thereafter. Despite decreasing TN loads, the worst hypoxic event ever recorded in open waters occurred in 2002. For estuaries and coastal areas, bottom water oxygen concentrations were best described by TN input from land and wind speed in July-September, explaining 52% of the interannual variation in concentrations. For open sea areas, bottom water oxygen concentrations were also modulated by TN input from land, however, additional significant variables included advective transport of water and Skagerrak surface water temperature and explained 49% of interannual variations in concentrations. Reductions in benthic species number and alpha diversity were significantly related to the duration of the 2002 hypoxic event. Gradual decreases in diversity measures (species number and alpha diversity) over the first 2-4 weeks show that the benthic community undergoes significant changes before the duration of hypoxia is severe enough to cause the community to collapse. Enhanced sediment-water fluxes of NH4+ and PO43- occur with hypoxia, increasing nutrient concentrations in the water column, and stimulating additional phytoplankton production. Repeated hypoxic events have changed the character of benthic communities and how organic matter is processed in sediments. Our data suggest that repeated hypoxic events lead to an increase in susceptibility of Danish waters to eutrophication and further hypoxia. (Less)

Denitrification measurements in aquatic sediments: A comparison of three methods

Measurements of denitrification using the acetylene inhibition,15N isotope tracer, and N2 flux methods were carried out concurrently using sediment cores from Vilhelmsborg sø, Denmark, in an attempt to clarify some of the limitations of each technique. Three experimental treatments of overlying water were used: control, nitrate enriched, and ammonia enriched water. The N2 flux and15N tracer experiments showed high rates of coupled nitrification/denitrification in the sediments. The acetylene inhibition method did not capture any coupled nitrification/denitrification. This could be explained by acetylene inhibition of nitrification. A combined15N tracer/acetylene inhibition experiment demonstrated that acetylene inhibition of N2O reduction was incomplete and the method, therefore, only measured approximately 50% of the denitrification due to nitrate from the overlying water. Similar rates of denitrification due to nitrate in the overlying water were measured by the N2 flux method and the acetylene inhibition method, after correcting for the 50% efficiency of acetylene inhibition. Rates of denitrification due to nitrate from the overlying water measured by the15N tracer method, however, were only approximately 35% or less of those measured by the acetylene inhibition or N2 flux methods.

Comparison of isotope pairing and N 2 /Ar methods for measuring sediment dentrification - assumptions, modifications and implications

Denitrification has been measured during the last few years using two different methods in particular: isotope pairing measured on a triple-collector isotopic ratio mass spectrometer and N2:Ar ratios measured on a membrane inlet mass spectrometer (MIMS). This study compares these two techniques in short-term batch experiments. Rates obtained using the original N2∶Ar method were up to 3 to 4 times higher than rates obtained using the isotope pairing technique due to O2 reacting with the N2 during MIMS analysis. Oxygen combines with N2 within the mass spectrometer ion source forming NO+ which reduces the N2 concentration. The decrease in N2 is least at lower O2 concentrations and since oxygen is typically consumed during incubations of sediment cores, the result is often a pseudo-increase in N2 concentration being interpreted as denitrification activity. The magnitude of this ocygen effect may be instrument specific. The reaction of O2 with N2 and the subsequent decrease in N2 was only partly correctly using an O2 correction curve for the relationship between N2 and O2 concentrations. The O2 corrected N2∶Ar denitrification rates were lower, but still did not match the isotope pairing rates and the variability between replicates was much higher. Using a copper reduction column heated to 600°C to remove all of the O2 from the sample before MIMS analysis resulted in comparable rates (slightly lower), and comparable variability between replicates, to the isotope pairing technique. The N2:Ar technique determines the net N2 production as the difference between N2 production by denitrification and N2 consumption by N-fixation, while N-fixation has little effect on the isotope pairing technique which determines a rate very close to the gross N2 production. When the two different techniques were applied on the same sediment, the small difference in rates obtained by the two methods seemed to reflect N-fixation as also supported from measurements of ethylene production in acetylene enriched sediment cores. The N2:Ar and isotope pairing techniques may be combined to provide simultaneous measurements of denitrification and N-fixation. Both techniques have several assumptions that must be met to achieve accurate rates; a number of tests are outlined that can be applied to demonstrate that these assumptions are being meet.

Impacts of longline mussel farming on oxygen and nitrogen dynamics and biological communities of coastal sediments

Benthic communities and benthic mineralization were studied in two shallow coastal regions of New Zealand: Tasman Bay, a possible future site for mussel farm development, and Beatrix Bay, which already hosts several longline mussel farms. In Tasman Bay, microphytobenthic (MPB) production added significantly to the total primary production of the bay. The activity of benthic microalgae had a pronounced effect on oxic conditions, solute exchange and denitrification rates. Benthic mineralization, quantified as the dark oxygen uptake, was in the range of 675±11 μmol m−2 h−1. Denitrification rates were high and fueled entirely by nitrate produced by the nitrifying community within the sediment. Competition for inorganic nitrogen between benthic microalgae and nitrifiers/denitrifiers resulted in diel variation in nitrogen cycling and reduced the inorganic nitrogen efflux and denitrification activity in the light. Calculated in electron equivalents, denitrification accounted for 11–20% of the total carbon mineralization—one of the highest numbers reported for coastal sediments. Reduced sediments, containing low MPB biomass and few subsurface macroinvertebrate species, were observed below a mussel farm in Beatrix Bay, presumably due to the intensified sedimentation of organic matter. Oxygen consumption increased in the organic-rich sediments, and ammonium effluxes were up to 14 times higher than those of unaffected sediments 250 m away from the farm. Denitrification rates below the farm were low as the coupled nitrification–denitrification was inhibited by the presence of sulfide. The dissimilative reduction of nitrate to ammonium (DRNA) was, however, stimulated in the reduced sediment. The enhanced benthic mineralization was associated with sulfidic sediments and a lower nitrogen removal rate due to impeded benthic photosynthesis and denitrification activity. The described local conditions associated with mussel farming should be taken into account when new areas are considered for development.

Denitrification and oxygen respiration in biofilms studied with a microsensor for nitrous oxide and oxygen

Depth distributions of O2 respiration and denitrification activity were studied in 1- to 2-mm thick biofilms from nutrient-rich Danish streams. Acetylene was added to block the reduction of N2O, and micro-profiles of O2 and N2O in the biofilm were measured simultaneously with a polarographic microsensor. The specific activities of the two respiratory processes were calculated from the microprofiles using a one-dimensional diffusion-reaction model. Denitrification only occurred in layers where O2 was absent or present at low concentrations (of a fewμM). Introduction of O2 into deeper layers inhibited denitrification, but the process started immediately after anoxic conditions were reestablished. Denitrification activity was present at greater depth in the biofilm when the NO3− concentration in the overlying water was elevated, and the deepest occurrence of denitrification was apparently determined by the depth penetration of NO3−. The denitrification rate within each specific layer was not affected by an increase in NO3− concentration, and the half-saturation concentration (Km) for NO3− therefore considered to be low (<25μM). Addition of 0.2% yeast extract stimulated denitrification only in the uppermost 0.2 mm of the denitrification zone indicating a very efficient utilization of the dissolved organic matter within the upper layers of the biofilm.

Microsensor Analysis of Oxygen in the Rhizosphere of the Aquatic Macrophyte Littorella uniflora (L.) Ascherson

Oxygen released by the roots of submerged plants may oxidize organic compounds from the roots and reduced substances continuously supplied by diffusion from the surrounding anoxic hydrosoil. We provide here the first visualization of this gradient environment obtained by microsensor analysis of oxygen in the rhizosphere of the freshwater plant Littorella uniflora (L.) Ascherson. The plants were rooted in an agar medium, in which amorphous FeS provided the main oxygen sink. The oxygen concentration at the root surface ranged from 20 to 450 [mu]M (atmospheric saturation = 280 [mu]M) between darkness and saturating light, and the oxic shell surrounding the roots varied from about 0.5 to 5 mm in thickness. The oxygen flux from the roots was a saturating function of the incident light intensity on the leaves, and the oxygen released was consumed mainly at the fluctuating oxic/anoxic interface. The oxic zones around individual roots are under dynamic control by light, root morphology, root density, and sediment reducing capacity, and, therefore, oxygen concentrations should be subject to substantial diurnal fluctuations in dense Littorella populations in nutrient-poor sediments.

Impact of Bacterial NO3− Transport on Sediment Biogeochemistry

ABSTRACT Experiments demonstrated that Beggiatoa could induce a H2S-depleted suboxic zone of more than 10 mm in marine sediments and cause a divergence in sediment NO3− reduction from denitrification to dissimilatory NO3− reduction to ammonium. pH, O2, and H2S profiles indicated that the bacteria oxidized H2S with NO3− and transported S0 to the sediment surface for aerobic oxidation.

Patterns of ammonium uptake within dense mats of the filamentous macroalga Chaetomorpha linum

Abstract The effect of macroalgal uptake on the flux of ammonium across the sediment-water interface was tested in laboratory experiments in which dense mats of Chaetomorpha linum were incubated at high and low surface irradiances and were exposed to a high simulated sediment nutrient flux. Depth profiles of NH + 4 concentrations within the 15-cm deep mats and the timing and magnitude of NH + 4 efflux through the mats to the overlying water reflected differences in macroalgal uptake between the two light treatments. Patterns of algal productivity and NH + 4 uptake with depth in the mats were determined from the accumulation of 13 C and 15 N in the algal tissue. Nitrogen-saturated macroalgae incubated at low irradiance exhibited a strong diel periodicity in NH + 4 uptake that was not present in the N-limited macroalgae incubated at high irradiance. Assimilation by the macroalgal mat at high irradiance was approximately 900 NH + 4 μmol m −2 h −1 , and was sufficient to prevent NH + 4 diffusion from the benthic nutrient source into the overlying water during both the light and dark periods. Uptake of NH + 4 in excess of the N growth demand in the lower half of the high-light mat resulted in a spatial separation of nutrient and light resources; NH + 4 did not diffuse into the upper layers and the most photosynthetically-active macroalgae remained N-deficient. Reduced irradiance decreased the total uptake of the mat by more than 50% (400 NH + 4 μmol m −2 h −1 ), and an efflux of NH + 4 into the overlying water occurred in the dark and early part of the light period. Ammonium diffused through the unproductive bottom layers of the low-light mat and was incorporated primarily in the photic zone in the upper 4 cm of the mat where photosynthesis provided the carbon required for N uptake and assimilation. These results support the hypothesis that actively-growing macroalgal mats efficiently sequester benthic nutrient inputs to the overlying water and reduce nutrient availability to a level that may limit pelagic production. Factors that reduce irradiance within the mat, such as self-shading or decreased insolation, limit macroalgal uptake of benthic flux and result in a release of nutrients into the overlying water.