Caitlin H. Frame

Assessment of Nitrogen and Oxygen Isotopic Fractionation During Nitrification and Its Expression in the Marine Environment

Nitrification is a microbially-catalyzed process whereby ammonia (NH(3)) is oxidized to nitrite (NO(2)(-)) and subsequently to nitrate (NO(3)(-)). It is also responsible for production of nitrous oxide (N(2)O), a climatically important greenhouse gas. Because the microbes responsible for nitrification are primarily autotrophic, nitrification provides a unique link between the carbon and nitrogen cycles. Nitrogen and oxygen stable isotope ratios have provided insights into where nitrification contributes to the availability of NO(2)(-) and NO(3)(-), and where it constitutes a significant source of N(2)O. This chapter describes methods for determining kinetic isotope effects involved with ammonia oxidation and nitrite oxidation, the two independent steps in the nitrification process, and their expression in the marine environment. It also outlines some remaining questions and issues related to isotopic fractionation during nitrification.

N2O production in the eastern South Atlantic: Analysis of N2O stable isotopic and concentration data

The stable isotopic composition of dissolved nitrous oxide (N2O) is a tracer for the production, transport, and consumption of this greenhouse gas in the ocean. Here we present dissolved N2O concentration and isotope data from the South Atlantic Ocean, spanning from the western side of the mid-Atlantic Ridge to the upwelling zone off the southern African coast. In the eastern South Atlantic, shallow N2O production by nitrifier denitrification contributed a flux of isotopically depleted N2O to the atmosphere. Along the African coast, N2O fluxes to the atmosphere of up to 46 µmol/m2/d were calculated using satellite-derived QuikSCAT wind speed data, while fluxes at the offshore stations averaged 0.04 µmol/m2/d. Comparison of the isotopic composition of the deeper N2O in the South Atlantic (800 m to 1000 m) to measurements made in other regions suggests that water advected from one or more of the major oxygen deficient zones contributed N2O to the mesopelagic South Atlantic via the Southern Ocean. This deeper N2O was isotopically and isotopomerically enriched (δ15Nbulk − N2O = 8.7 ± 0.1‰, δ18O − N2O = 46.5 ± 0.2‰, and Site Preference = 18.7 ± 0.6‰) relative to the shallow N2O source, indicating that N2O consumption by denitrification influenced its isotopic composition. The N2O concentration maximum was observed between 200 m and 400 m and reached 49 nM near the Angolan coast. The depths of the N2O concentration maximum coincided with those of sedimentary particle resuspension along the coast. The isotopic composition of this N2O (δ15Nbulk − N2O = 5.8 ± 0.1‰, δ18O − N2O = 39.7 ± 0.1‰, and Site Preference = 9.8 ± 1.0‰) was consistent with production by diffusion-limited nitrate (NO3−) reduction to nitrite (NO2−), followed by NO2− reduction to N2O by denitrification and/or nitrifier denitrification, with additional N2O production by NH2OH decomposition during NH3 oxidation. The sediment surface, benthic boundary layer, or particles resuspended from the sediments are likely to have provided the physical and chemical conditions necessary to produce this N2O.

Acidification Enhances Hybrid N2O Production Associated with Aquatic Ammonia-Oxidizing Microorganisms

Ammonia-oxidizing microorganisms are an important source of the greenhouse gas nitrous oxide (N2O) in aquatic environments. Identifying the impact of pH on N2O production by ammonia oxidizers is key to understanding how aquatic greenhouse gas fluxes will respond to naturally occurring pH changes, as well as acidification driven by anthropogenic CO2. We assessed N2O production rates and formation mechanisms by communities of ammonia-oxidizing bacteria (AOB) and archaea (AOA) in a lake and a marine environment, using incubation-based nitrogen (N) stable isotope tracer methods with 15N-labeled ammonium (15NH4+) and nitrite (15NO2−), and also measurements of the natural abundance N and O isotopic composition of dissolved N2O. N2O production during incubations of water from the shallow hypolimnion of Lake Lugano (Switzerland) was significantly higher when the pH was reduced from 7.54 (untreated pH) to 7.20 (reduced pH), while ammonia oxidation rates were similar between treatments. In all incubations, added NH4+ was the source of most of the N incorporated into N2O, suggesting that the main N2O production pathway involved hydroxylamine (NH2OH) and/or NO2− produced by ammonia oxidation during the incubation period. A small but significant amount of N derived from exogenous/added 15NO2− was also incorporated into N2O, but only during the reduced-pH incubations. Mass spectra of this N2O revealed that NH4+ and 15NO2− each contributed N equally to N2O by a “hybrid-N2O” mechanism consistent with a reaction between NH2OH and NO2−, or compounds derived from these two molecules. Nitrifier denitrification was not an important source of N2O. Isotopomeric N2O analyses in Lake Lugano were consistent with incubation results, as 15N enrichment of the internal N vs. external N atoms produced site preferences (25.0–34.4‰) consistent with NH2OH-dependent hybrid-N2O production. Hybrid-N2O formation was also observed during incubations of seawater from coastal Namibia with 15NH4+ and NO2−. However, the site preference of dissolved N2O here was low (4.9‰), indicating that another mechanism, not captured during the incubations, was important. Multiplex sequencing of 16S rRNA revealed distinct ammonia oxidizer communities: AOB dominated numerically in Lake Lugano, and AOA dominated in the seawater. Potential for hybrid N2O formation exists among both communities, and at least in AOB-dominated environments, acidification may accelerate this mechanism.

Powering up the “biogeochemical engine”: the impact of exceptional ventilation of a deep meromictic lake on the lacustrine redox, nutrient, and methane balances

The Lake Lugano North Basin has been meromictic for several decades, with anoxic waters below 100m depth. Two consecutive cold winters in 2005 and 2006 induced exceptional deep mixing, leading to a transient oxygenation of the whole water column. With the ventilation of deep waters and the oxidation of large quantities of reduced solutes, the lakes total redox-balance turned positive, and the overall hypolimnetic oxygen demand of the lake strongly decreased. The disappearance of 150 t dissolved phosphorous (P) during the first ventilation in March 2005 is attributed to the scavenging of water-column-borne P by newly formed metal oxyhydroxides and the temporary transfer to the sediments. The fixed nitrogen (N) inventory was reduced by ~30% (~1000 t). The water-column turnover induced the nitratation of the previously NO3--free deep hypolimnion by oxidation of large amounts of legacy NH4+ and by mixing with NO3--rich subsurface water masses. Sediments with a strong denitrifying potential, but NO3--starved for decades, were brought in contact with NO3--replete waters, invigorating benthic denitrification and rapid fixed N loss from the lake in spite of the overall more oxygenated conditions. Similarly, a large microbial aerobic CH4 oxidation (MOx) potential in the hypolimnion was capitalized with the ventilation of the deep basin. Almost all CH4, which had been built up over more than 40 years (~2800 t), was removed from the water column within 30 days. However, boosted MOx could only partly explain the disappearance of the CH4. The dominant fraction (75%) of the CH4 evaded to the atmosphere, through storage flux upon exposure of anoxic CH4-rich water to the atmosphere. As of today, the North Basin seems far from homeostasis regarding its fixed N and CH4 budgets, and the deep basins CH4 pool is recharging at a net production rate of ~66 t y-1. The size of impending CH4 outbursts will depend on the frequency and intensity of exceptional mixing events in the future.