Description: Upwelling systems are significant sources of atmospheric nitrous oxide (N₂O). The Benguela Upwelling System is one of the most productive regions worldwide and a temporally variable source of N₂O. Strong O₂ depletions above the shelf are favoring periodically OMZ formations. We aimed to assess underlying N₂O production and consumption processes on different temporal and spatial scales during austral winter in the Benguela Upwelling System, when O₂⁻deficiency in the water column is relatively low. The fieldwork took place during the cruise M157 (August 4ᵗʰ – September 16ᵗʰ 2019) onboard the R/V METEOR. This expedition included four close-coastal regions around Walvis Bay at 23°S, which presented the lowest O₂ concentrations near the seafloor and thus may provide hotspots of N₂O production. Seawater was collected in 10 L free-flow bottles by using a rosette system equipped with conductivity-temperature-depth (CTD) sensors (SBE 911plus, Seabird-electronics, USA).Seawater samples were collected from 10 L free-flow bottles bubble-free, filled into 200 mL serum bottles and immediately fixed with saturated mercury chloride (HgCl₂). Concentrations of dissolved N₂O were measured by a purge and trap system using a dynamic headspace (Sabbaghzadeh et al., 2021). The N₂O gas saturation (N₂Oₛₐₜ in %) was calculated from the concentration ratio between the seawater sample and seawater equilibrated with the atmosphere. ∆N₂O (N₂O saturation disequilibrium in nmol L⁻¹) was calculated as the difference between the measured N₂O concentration and the atmospheric equilibrium N₂O concentration using Bunsen solubility coefficient (Weiss and Price, 1980). AOU (apparent oxygen utilization in µmol L⁻¹) expresses the O₂ consumption by microbial respiration and was calculated as the difference between the equilibrated O₂ and observed O₂ concentration with the same physico-chemical properties (Weiss and Price, 1980).

Description: Upwelling systems are significant sources of atmospheric nitrous oxide (N₂O). The Benguela Upwelling System is one of the most productive regions worldwide and a temporally variable source of N₂O. Strong O₂ depletions above the shelf are favoring periodically OMZ formations. We aimed to assess underlying N₂O production and consumption processes on different temporal and spatial scales during austral winter in the Benguela Upwelling System, when O₂-deficiency in the water column is relatively low. The fieldwork took place during the cruise M157 (August 4th – September 16th 2019) onboard the R/V METEOR. This expedition included four close-coastal regions around Walvis Bay at 23°S, which presented the lowest O₂ concentrations near the seafloor and thus may provide hotspots of N₂O production. Seawater was collected in 10 L free-flow bottles by using a rosette system equipped with conductivity-temperature-depth (CTD) sensors (SBE 911plus, Seabird-electronics, USA). Incubation experiments were performed using stable isotope ¹⁵N-tracers. Seawater samples for ¹⁵N-tracer incubations and natural abundance N₂O analysis were collected from 10 L free-flow bottles and filled bubble-free into 125 mL serum bottles. The samples for natural abundance N₂O analysis were immediately fixed with saturated HgCl₂ and stored in the dark. To perform the incubation, we added ¹⁵N-labeled NO₂-, NO₃⁻ and NH₄⁺ to estimate the in-situ N₂O production rates and associated reactions. To determine a single rate, the bottles were sacrificed after tracer addition, and within the time interval of 12 h, 24 h and 48 h by adding HgCl₂. Rates were calculated based on a linear regression over time. Total N₂O and natural abundance isotopologues of N₂O were analyzed by using an isotope ratio mass spectrometer (IRMS, Delta V Plus, Thermo Scientific). NO₂- production was additionally analyzed by transforming ¹⁵NO₂- to ¹⁵N₂O following the azide method after McIlvin & Altabet (2005) and the nitrogen isotope ratio of N₂O was measured by an IRMS. N₂ production was determined via an IRMS (Flash-EA-ConfloIV-DELTA V Advanced, Thermo Scientific) by injecting headspace from exetainers. The N₂O yield per nitrite produced and the N₂O yield during denitrification was calculated. Samples for natural abundance N₂O was sampled and measured in triplicates and is shown as an average with standard deviation (SD). In order to estimate the contribution of different N₂O producing pathways by major biological processes and the extent of N₂O reduction to N₂, the dual-isotope mapping approach was applied to natural abundance isotopologues of N₂O, which uses the relative position of background-subtracted N₂O samples in a δ¹⁵Nˢᴾ-N₂O vs. δ¹⁸O-N₂O diagram (Yu et al., 2020; Lewicka-Szczebak et al., 2020).

Description: Upwelling systems are significant sources of atmospheric nitrous oxide (N₂O). The Benguela Upwelling System is one of the most productive regions worldwide and a temporally variable source of N₂O. Strong O₂ depletions above the shelf are favoring periodically OMZ formations. We aimed to assess underlying N₂O production and consumption processes on different temporal and spatial scales during austral winter in the Benguela Upwelling System, when O₂-deficiency in the water column is relatively low. The fieldwork took place during the cruise M157 (August 4th – September 16th 2019) onboard the R/V METEOR. This expedition included four close-coastal regions around Walvis Bay at 23°S, which presented the lowest O₂ concentrations near the seafloor and thus may provide hotspots of N₂O production. Seawater was collected in 10 L free-flow bottles by using a rosette system equipped with conductivity-temperature-depth (CTD) sensors (SBE 911plus, Seabird-electronics, USA). Incubation experiments were performed using stable isotope ¹⁵N-tracers. Seawater samples for ¹⁵N-tracer incubations and natural abundance N₂O analysis were collected from 10 L free-flow bottles and filled bubble-free into 125 mL serum bottles. The samples for natural abundance N₂O analysis were immediately fixed with saturated HgCl₂ and stored in the dark. To perform the incubation, we added ¹⁵N-labeled NO₂-, NO₃⁻ and NH₄⁺ to estimate the in-situ N₂O production rates and associated reactions. To determine a single rate, the bottles were sacrificed after tracer addition, and within the time interval of 12 h, 24 h and 48 h by adding HgCl₂. Rates were calculated based on a linear regression over time. Total N₂O and natural abundance isotopologues of N₂O were analyzed by using an isotope ratio mass spectrometer (IRMS, Delta V Plus, Thermo Scientific). NO₂- production was additionally analyzed by transforming ¹⁵NO₂- to ¹⁵N₂O following the azide method after McIlvin & Altabet (2005) and the nitrogen isotope ratio of N₂O was measured by an IRMS. N₂ production was determined via an IRMS (Flash-EA-ConfloIV-DELTA V Advanced, Thermo Scientific) by injecting headspace from exetainers. The N₂O yield per nitrite produced and the N₂O yield during denitrification was calculated. Samples for natural abundance N₂O was sampled and measured in triplicates and is shown as an average with standard deviation (SD). In order to estimate the contribution of different N₂O producing pathways by major biological processes and the extent of N₂O reduction to N₂, the dual-isotope mapping approach was applied to natural abundance isotopologues of N₂O, which uses the relative position of background-subtracted N₂O samples in a δ¹⁵Nˢᴾ-N₂O vs. δ¹⁸O-N₂O diagram (Yu et al., 2020; Lewicka-Szczebak et al., 2020).

Description: Upwelling systems are significant sources of atmospheric nitrous oxide (N₂O). The Benguela Upwelling System is one of the most productive regions worldwide and a temporally variable source of N₂O. Strong O₂ depletions above the shelf are favoring periodically OMZ formations. We aimed to assess underlying N₂O production and consumption processes on different temporal and spatial scales during austral winter in the Benguela Upwelling System, when O₂-deficiency in the water column is relatively low. The fieldwork took place during the cruise M157 (August 4ᵗʰ – September 16ᵗʰ 2019) onboard the R/V METEOR. This expedition included four close-coastal regions around Walvis Bay at 23°S, which presented the lowest O₂ concentrations near the seafloor and thus may provide hotspots of N₂O production. Seawater was collected in 10 L free-flow bottles by using a rosette system equipped with conductivity-temperature-depth (CTD) sensors (SBE 911plus, Seabird-electronics, USA). Concentrations of inorganic nutrients (PO₄³⁻, NH₄⁺, NO₃⁻, NO₂⁻, and SiO₂) were measured colorimetrically according to Grasshoff et al. (1999) by means of a continuous segmented flow analyzer (SEAL Analytical, QuAAtro39). To determine the water mass fractions along the sampling transects, vertical profiles were collected using a free-falling microstructure profiler (MSS90L, Sea & Sun Technology). Temperature, dissolved oxygen, and salinity were measured with a CTD system consisting of a SeaBird 911+ probe, mounted on a sampling rosette.

Description: In March/April 2018 during a cruise on R/V Sally Ride, SR1805, 15N-NH4+ incubations in 60mL glass serum bottles were performed to measure ammonium oxidation rates to nitrite and nitrous oxide in different depth at 3 different stations in the oxygen deficient zone (ODZ) of the Eastern Tropical North Pacific off the coast of Mexico. Water samples were collected from 30L Niskin bottles deployed with a conductivity-temperature-depth profiler (CTD, Seabird Electronics). The goal was to get a better understanding on the controls of nitrous oxide (N2O) production. The N2O production rate experiments were performed according to Bourbonnais et al. 2021 (https://doi.org/10.3389/fmars.2021.611937). Furthermore, ammonium (NH4+), nitrite (NO2-) and nitrate (NO3-) as well as N2O concentrations were determined using standard fluorometric (Holmes et al. 1999, https://doi.org/10.1139/f99-128), photometric (Strickland and Parsons 1972, hdl:10013/epic.46454.d001), chemiluminescent (Braman and Hendrix 1989, doi:10.1021/ac00199a007) and mass spectrometric techniques (McIlvin and Casciotti 2010, https://doi.org/10.4319/lom.2010.8.54), respectively. The N2O yield per nitrite produced was calculated. The archaeal ammonia monooxygenase gene subunit A (amoA) copy numbers/mL were determined using qPCR as described previously (Peng et al. 2015, https://doi.org/10.1002/2015GB005278).

Description: The Kryos Basin is a deep-sea hypersaline anoxic basin (DHAB) located in the Eastern Mediterranean Sea (34.98°N 22.04°E). It is filled with brine of re-dissolved Messinian evaporites and is nearly saturated with MgCl2-equivalents, which makes this habitat extremely challenging for life. The strong density difference between the anoxic brine and the overlying oxic Mediterranean seawater impedes mixing, giving rise to a narrow chemocline. Here, we investigate the microbial community structure and activities across the seawater–brine interface using a combined biogeochemical, next-generation sequencing, and lipid biomarker approach. Within the interface, we detected fatty acids that were distinctly 13C-enriched when compared to other fatty acids. These likely originated from sulfide-oxidizing bacteria that fix carbon via the reverse tricarboxylic acid cycle. In the lower part of the interface, we also measured elevated rates of methane oxidation, probably mediated by aerobic methanotrophs under micro-oxic conditions. Sulfate reduction rates increased across the interface and were highest within the brine, providing first evidence that sulfate reducers (likely Desulfovermiculus and Desulfobacula) thrive in the Kryos Basin at a water activity of only ~0.4 Aw. Our results demonstrate that a highly specialized microbial community in the Kryos Basin has adapted to the poly-extreme conditions of a DHAB with nearly saturated MgCl2 brine, extending the known environmental range where microbial life can persist.

Description: Over much of the ocean’s surface, productivity and growth are limited by a scarcity of bioavailable nitrogen. Sedimentary δ15N records spanning the last deglaciation suggest marked shifts in the nitrogen cycle during this time, but the quantification of these changes has been hindered by the complexity of nitrogen isotope cycling. Here we present a database of δ15N in sediments throughout the world’s oceans, including 2,329 modern seafloor samples, and 76 timeseries spanning the past 30,000 years. We show that the δ15N values of modern seafloor sediments are consistent with values predicted by our knowledge of nitrogen cycling in the water column. Despite many local deglacial changes, the globally averaged δ15N values of sinking organic matter were similar during the Last Glacial Maximum and Early Holocene. Considering the global isotopic mass balance, we explain these observations with the following deglacial history of nitrogen inventory processes. During the Last Glacial Maximum, the nitrogen cycle was near steady state. During the deglaciation, denitrification in the pelagic water column accelerated. The flooding of continental shelves subsequently increased denitrification at the seafloor, and denitrification reached near steady-state conditions again in the Early Holocene. We use a recent parameterization of seafloor denitrification to estimate a 30–120% increase in benthic denitrification between 15,000 and 8,000 years ago. Based on the similarity of globally averaged δ15N values during the Last Glacial Maximum and Early Holocene, we infer that pelagic denitrification must have increased by a similar amount between the two steady states.

Description: We present a new nitrogen isotope model incorporated into the three-dimensional ocean component of a global Earth system climate model designed for millennial timescale simulations. The model includes prognostic tracers for the two stable nitrogen isotopes, 14N and 15N, in the nitrate (NO3−), phytoplankton, zooplankton, and detritus variables of the marine ecosystem model. The isotope effects of algal NO3− uptake, nitrogen fixation, water column denitrification, and zooplankton excretion are considered as well as the removal of NO3− by sedimentary denitrification. A global database of δ15NO3− observations is compiled from previous studies and compared to the model results on a regional basis where sufficient observations exist. The model is able to qualitatively and quantitatively reproduce many of the observed patterns such as high subsurface values in water column denitrification zones and the meridional and vertical gradients in the Southern Ocean. The observed pronounced subsurface minimum in the Atlantic is underestimated by the model presumably owing to too little simulated nitrogen fixation there. Sensitivity experiments reveal that algal NO3− uptake, nitrogen fixation, and water column denitrification have the strongest effects on the simulated distribution of nitrogen isotopes, whereas the effect from zooplankton excretion is weaker. Both water column and sedimentary denitrification also have important indirect effects on the nitrogen isotope distribution by reducing the fixed nitrogen inventory, which creates an ecological niche for nitrogen fixers and, thus, stimulates additional N2 fixation in the model. Important model deficiencies are identified, and strategies for future improvement and possibilities for model application are outlined.