While the ocean's photosynthetic production of organic matter rivals that on land, a combination of heterotrophy and sinking prevents significant accumulation of particulate organic matter (POM) in open ocean surface waters. The origins and fates of POM in ocean surface waters are unclear, in part due to the dominance of nonliving, altered material. From the natural nitrogen isotopic composition of chlorophyll and its degradation products, we estimate the fraction of particles from eukaryotic vs. prokaryotic phytoplankton. In subtropical gyres and along the eastern North Pacific margin, the eukaryotic-to-prokaryotic ratio in particles matches that of living phytoplankton. However, in the North Atlantic outside its subtropical gyre, particles have a lower eukaryotic-to-prokaryotic ratio than do the living phytoplankton. This discrepancy at least partly arises from preferential sinking of eukaryotic biomass, consistent with the canonical but disputed paradigm that cyanobacteria disproportionately fulfill the energetic demands of the upper ocean microbial community while eukaryotes drive export production. The prokaryotic bias in surface ocean particles may also result from slow decomposition of specific components of prokaryotic biomass, a possible bottleneck in the ocean's microbial loop. The different fates of organic matter produced by eukaryotic and prokaryotic phytoplankton affect the productivity of the surface ocean, carbon export to the interior, and the signals recorded in deep-sea sediments.

The physical and biogeochemical properties of the western Arctic Ocean are rapidly changing, resulting in cascading shifts to the local ecosystems. The nutrient‐rich Pacific water inflow to the Arctic through the Bering Strait is modified on the Chukchi and East Siberian shelves by brine rejection during sea ice formation, resulting in a strong halocline (called the Upper Halocline Layer (UHL)) that separates the cold and relatively fresh surface layer from the warmer and more saline (and nutrient‐poor) Atlantic‐derived water below. Biogeochemical signals entrained into the UHL result from Pacific Waters modified by sediment and river influence on the shelf. In this synthesis, we bring together data from the 2015 Arctic U.S. GEOTRACES program to implement a multi‐tracer (dissolved and particulate trace elements, radioactive and stable isotopes, macronutrients, and dissolved gas/atmospheric tracers) approach to assess the relative influence of shelf sediments, rivers, and Pacific seawater contribution to the Amerasian Arctic halocline. For each element, we characterized their behavior as mixing dominated (e.g., dCu, dGa), shelf‐influenced (e.g., dFe, dZn), or a combination of both (e.g., dBa, dNi). Leveraging this framework, we assessed sources and sinks contributing to elemental distributions: shelf sediments (e.g., dFe, dZn, dCd, dHg), riverine sources, (e.g., dCu, dBa, dissolved organic carbon), and scavenging by particles originating on the shelf (e.g., dFe, dMn, dV, etc.). Additionally, synthesized results from isotopic and atmospheric tracers yielded tracer age estimates for the Upper Halocline ranging between 1 and 2 decades on a spatial gradient consistent with cyclonic circulation.

The ocean accounts for ∼20%–30% of global nitrous oxide (N 2 O) emissions, with coastal upwelling systems estimated to contribute disproportionately to the sea‐air flux of this potent greenhouse gas. To investigate the mechanisms of and controls on N 2 O production in coastal upwelling systems, we measured the concentration and nitrogen and oxygen isotopic composition of N 2 O (δ 15 N‐N 2 O and δ 18 O‐N 2 O) along a cross‐shelf transect in the Southern Benguela Upwelling System (SBUS). At the shelf bottom, N 2 O concentrations increased from the outer shelf toward the shore (11–32 nM) inversely to dissolved oxygen (182 ± 17 to <1 μM) and in concert with the remineralization tracers, apparent oxygen utilization (108 ± 21 to 221 ± 33 μM) and nitrogen (N)‐deficit (up to 20.4 μM). These observations suggest that both nitrification and denitrification may be involved in N 2 O production on the SBUS shelf. The δ 15 N‐N 2 O implicates both processes as potential N 2 O sources on the shelf, with high δ 18 O‐N 2 O values (≤57.2‰) specifically incriminating sediments as the primary N 2 O source to the water column. Isotopic changes across the shelf delineate three discrete domains with distinct N 2 O sources. Sedimentary nitrification and/or denitrification dominate N 2 O production on the midshelf , while coupled nitrification‐denitrification explains N 2 O production on the inner‐shelf . At the shallow inner‐shelf where oxygen is depleted, both water column and sedimentary denitrification account for the production and partial consumption of N 2 O. This study illuminates the disproportionate contribution of sedimentary N cycling to N 2 O production on the SBUS shelf.

St Helena Bay (SHB), a retentive zone in the productive southern Benguela Upwelling System off western South Africa, experiences seasonal hypoxia and episodic anoxic events that threaten local fisheries. To understand the drivers of oxygen variability in SHB, we queried 25 years of dissolved oxygen (DO) observations alongside high‐resolution wind and hydrographic data, and dynamical data from a high‐resolution model. At 70 m in SHB (mid‐bay), upwelling‐favorable winds in spring drove replenishment of cold, oxygenated water. Hypoxia developed in summer, becoming most severe in autumn. Bottom waters in autumn were replenished with warmer, less oxygenated water than in spring—suggesting a seasonal change in source waters upwelled into the bay. Downwelling and deep mixing in winter ventilated mid‐bay bottom waters, which reverted to hypoxic conditions during wind relaxations and reversals. In the nearshore (20 m), hypoxia occurred specifically during periods of upwelling‐favorable wind stress and was most severe in autumn. Using a statistical model, we extended basic hydrographic observations to nitrate and DO concentrations and developed metrics to identify the accumulation of excess nutrients on the shelf and nitrogen‐loss to denitrification, both of which were most prominent in autumn. A correspondence of the biogeochemical properties of hypoxic waters at 20 m to those at 70 m implicates the latter as the source waters upwelled inshore in autumn. We conclude that wind‐driven upwelling drives the replenishment of respired bottom waters in SHB with oxygenated waters, noting that less‐oxygenated water is imported later in the upwelling season, which exacerbates hypoxia.

We examined the nitrogen (N) biogeochemistry of adjacent cyclonic and anticyclonic eddies near Hawai’i in the North Pacific Subtropical Gyre (NPSG) and explored mechanisms that may sustain productivity in the cyclone after the initial intensification stage. The top of the nutricline was uplifted into the euphotic zone in the cyclone and depressed in the anticyclone. Subsurface nutrient concentrations and apparent oxygen utilization at the cyclone’s inner periphery were higher than expected from isopycnal displacement, suggesting that shallow remineralization of organic material generated excess nutrients in the subsurface. The excess nutrients may provide a supply of subsurface nutrients to sustain productivity in maturing eddies. The shallow remineralization also raises questions regarding the extent to which cyclonic eddies promote deep carbon sequestration in subtropical gyres such as the NPSG. An upward increase in nitrate 15N/14N isotope ratios below the euphotic zone, indicative of partial nitrate assimilation, coincided with negative preformed nutrients – potentially signaling heterotrophic bacterial consumption of carbon-rich (nitrogen-poor) organic material. The 15N/14N of material collected in shallow sediment traps was significantly higher in the cyclone than the anticyclone and showed correspondence to the 15N/14N ratio of the nitrate supply, which is acutely sensitive to sea level anomaly in the region. A number of approaches were applied to estimate the contribution of N2 fixation to export production; results among approaches were inconsistent, which we attribute to non-steady state conditions during our observation period.

Climate change is expected to alter the input of nitrogen (N) sources in the Eastern Canadian Arctic Archipelago (ECAA) and Baffin Bay due to increased discharge from glacial meltwater and permafrost thaw. Since dissolved inorganic N is generally depleted in surface waters, dissolved organic N (DON) could represent a significant N source fueling phytoplankton activity in Arctic ecosystems. Yet, few DON data for this region exist. We measured concentrations and stable isotope ratios (δ15N and δ18O) of DON and nitrate (NO3−) to investigate the sources and cycling of dissolved nitrogen in regional rivers and at the sea surface from samples collected in the ECAA and Baffin Bay during the summer of 2019. The isotopic signatures of NO3- in rivers could be reproduced in a steady state isotopic model by invoking mixing between atmospheric NO3- and nitrified ammonium as well as NO3- assimilation by phytoplankton. DON concentrations were low in most rivers (≤4.9 µmol L−1), whereas the concentrations (0.54–12 µmol L−1) and δ15N of DON (−0.71–9.6 ‰) at the sea surface were variable among stations, suggesting dynamic cycling and/or distinctive sources. In two regions with high chl-a, DON concentrations were inversely correlated with chlorophyll‐an and the d15N of DON, suggesting net DON consumption in localized phytoplankton blooms. We derived an isotope effect of −6.9‰ for DON consumption. Our data helps establish a baseline to assess future change in nutrient regime for this climate sensitive region.

Observations and model studies suggest that front dynamics can enhance phytoplankton productivity. This study tested whether frontal systems also increase the abundance of nitrifying microbes and nitrogen recycling during repeat sampling transects across the Mid-Atlantic Bight shelfbreak in July 2019. We measured ammonium concentrations, nitrate dual isotopes (delta 15N, delta 18O), and ammonia monooxygenase subunit A (amoA) genes of ammonia-oxidizing archaea (AOA) and bacteria (AOB). In subsurface shelf waters, ammonium concentrations exceeded 2 mu mol L-1, due to a temporary imbalance in regeneration from sinking particles and subsequent nitrification. The inverse correlation between nitrate delta 15N values and ammonium concentrations confirmed nitrate was partially or entirely from local nitrification on the shelf. In contrast, the shelfbreak frontal zone and slope sea subsurface waters had much lower ammonium concentrations (0.1-0.2 mu mol L-1) due to tight coupling between ammonium regeneration and nitrification. The deviation of nitrate delta 15N and delta 18O from algal uptake-driven 1 : 1 ratio suggests concurrent nitrification in the euphotic zone. The shelfbreak front acted as an ecological boundary where AOA and AOB amoA gene numbers were partitioned, with AOAs abounding in slope waters and AOBs in shelf waters, likely due to ammonium availability. At certain slope stations, deep-water nutrient inputs via isopycnal lifting induced by Gulf Stream intrusions caused unexpectedly high phytoplankton biomass, which doubled nitrifier abundance and potentially stimulated both ammonium regeneration and nitrification. These findings demonstrate distinct distributions of nitrifying microbes along the salinity gradient from shelf to slope and highlight the significant influence of coastal ocean-western boundary current interactions on nitrogen biogeochemistry.

The nitrogen (N) isotope composition (δ15N) of cold-water corals is a promising proxy for reconstructing past ocean N cycling, as a strong correlation was found between the δ15N of the organic nitrogen preserved in coral skeletons and the δ15N of particulate organic matter exported from the surface ocean. However, a large offset of 8 ‰–9 ‰ between the δ15N recorded by the coral and that of exported particulate organic matter remains unexplained. The 8 ‰–9 ‰ offset may signal a higher trophic level of coral dietary sources, an unusually large trophic isotope effect or a biosynthetic δ15N offset between the coral's soft tissue and skeletal organic matter, or some combinations of these factors. To understand the origin of the offset and further validate the proxy, we investigated the trophic ecology of the asymbiotic scleractinian cold-water coral Balanophyllia elegans, both in a laboratory setting and in its natural habitat. A long-term incubation experiment of B. elegans fed on an isotopically controlled diet yielded a canonical trophic isotope effect of 3.0 ± 0.1 ‰ between coral soft tissue and the Artemia prey. The trophic isotope effect was not detectably influenced by sustained food limitation. A long N turnover of coral soft tissue, expressed as an e-folding time, of 291 ± 15 d in the well-fed incubations indicates that coral skeleton δ15N is not likely to track subannual (e.g., seasonal) variability in diet δ15N. Specimens of B. elegans from the subtidal zone near San Juan Channel (WA, USA) revealed a modest difference of 1.2 ± 0.6 ‰ between soft tissue and skeletal δ15N. The δ15N of the coral soft tissue was 12.0 ± 0.6 ‰, which was ∼6 ‰ higher than that of suspended organic material that was comprised dominantly of phytoplankton – suggesting that phytoplankton is not the primary component of B. elegans' diet. An analysis of size-fractionated net tow material suggests that B. elegans fed predominantly on a size class of zooplankton ≥500 µm, implicating a two-level trophic transfer between phytoplankton material and coral tissue. These results point to a feeding strategy that may result in an influence of the regional food web structure on the cold-water coral δ15N. This factor should be taken into consideration when applying the proxy to paleo-oceanographic studies of ocean N cycling.

The cyclic growth and decay of continental ice sheets can be reconstructed from the history of global sea level. Sea level is relatively well constrained for the Last Glacial Maximum (LGM, 26,500 to 19,000 y ago, 26.5 to 19 ka) and the ensuing deglaciation. However, sea-level estimates for the period of ice-sheet growth before the LGM vary by > 60 m, an uncertainty comparable to the sea-level equivalent of the contemporary Antarctic Ice Sheet. Here, we constrain sea level prior to the LGM by reconstructing the flooding history of the shallow Bering Strait since 46 ka. Using a geochemical proxy of Pacific nutrient input to the Arctic Ocean, we find that the Bering Strait was flooded from the beginning of our records at 46 ka until 35.7 - 2.4 + 3.3 ka. To match this flooding history, our sea-level model requires an ice history in which over 50% of the LGM’s global peak ice volume grew after 46 ka. This finding implies that global ice volume and climate were not linearly coupled during the last ice age, with implications for the controls on each. Moreover, our results shorten the time window between the opening of the Bering Land Bridge and the arrival of humans in the Americas.

Abstract. The nitrogen (N) isotope composition (δ15N) of cold-water corals is a promising proxy for reconstructing past ocean N cycling, as a strong correlation was found between the δ15N of the organic nitrogen preserved in coral skeletons and the δ15N of sinking particulate organic matter exported from the surface ocean. However, a large offset of 8–9 ‰ between the δ15N recorded by the coral and that of export remains unexplained. The 8–9 ‰ offset may signal a potential sensitivity of the proxy to food web structure, an unusual large trophic isotope effect or a biosynthetic δ15N offset between the coral’s soft tissue and skeletal tissues, or some combinations of these factors. To understand the origin of the offset and further validate the proxy, we investigated the trophic ecology of the scleractinan cold water coral Balanophyllia elegans. A long-term incubation experiment of B. elegans fed on an isotopically controlled diet yielded a canonical trophic isotope effect of 3.0 ± 0.1 ‰ between coral soft tissue and the Artemia prey. The trophic isotope effect was not detectably influenced by sustained food limitation. A long N turnover of coral soft tissue of 291 ± 15 days in the well-fed incubations indicates that the coral skeleton is not apt to record seasonal difference in diet δ15N. Specimens of B. elegans from the shallow subtidal zone near San Juan Channel (WA, USA) revealed a modest difference between soft and skeletal tissue δ15N of 1.2 ± 0.6 ‰. The δ15N of the coral soft tissue was 12.0 ± 0.6 ‰, which was ~6 ‰ higher than that of suspended organic material that was comprised dominantly of phytoplankton – suggesting that the latter is not the primary component of B. elegans’ diet. An analysis of size-fractionated net tow material suggests that B. elegans fed predominantly on a size class of zooplankton ≥ 500 µm, implicating a two-level trophic transfer between phytoplankton material and coral tissue. These results portend a sensitivity of cold-water coral δ15N to regional food web structure that must be heeded in paleoceanographic studies of ocean N cycling.

Here we use data from three meridional survey cruises that took place in 1994, 2007/2008 and 2016/2017 in the southeast Pacific Ocean to investigate controls on surface distributions of dissolved organic carbon and nitrogen (DOC and DON, respectively). While DOC and DON production occur simultaneously in the euphotic layer of the gyre margins, budgets that account for horizontal transport, production and consumption are not balanced across the region. Our results show that while DOC concentrations increase inside the gyre, DON remains at similar levels in comparison to margins. There is little interannual variability in DOC and DON trends, except near the equator, explained by ENSO phases and seasonality. While the effect of evaporation (precipitation) did not significantly contribute to variable DOC and DON concentrations, differences in the naturally occurring N-15/N-14 isotope ratios from the most recent cruise suggest net consumption of allochthonous DON inside the gyre. In the northernmost and southernmost stations, where surface [NO3-] was detected, N-15-DON showed low signatures in the upper 300 m of 3.8 +/- 1.5%o and 2.5 +/- 1.6%o, respectively. Inside the gyre where [NO3-] was undetectable, higher values averaging 5.0 +/- 0.9%o were observed. This suggests that a fraction of DON turns over more rapidly than the time scale of horizontal transport, with the isotopic change reflecting the gradient in N-15-DON of newly produced organic matter. Thus, allochthonous DON is likely a significant source of nitrogen to microbial communities inside this nitrogen-limited environment and should be taken into consideration when closing nutrient budgets to estimate productivity in the South Pacific Subtropical Gyre.

The Canadian Arctic Archipelago (CAA) provides a gateway for the flow of nutrients from the North Pacific to the North Atlantic. The transport and biogeochemical cycling of nutrients throughout the CAA, however, are sparsely documented. Here, we report water column nitrogen and oxygen isotope ratios of nitrate (delta N-15(NO3) and delta O-18(NO3)) collected in a throughflow of the CAA into Baffin Bay, providing insights on inherent nutrient dynamics. The nitracline shoaled eastward into the CAA, wherein large subsurface chlorophyll maxima and coincident increases in delta N-15(NO3) and delta O-18(NO3) indicated enhanced nitrate assimilation relative to the oligotrophic Canada Basin. High delta N-15(NO3) values characteristic of Pacific Winter Water (PWW) pervaded the Archipelago and Baffin Bay, decreasing eastward due to mixing with underlying Atlantic water (AW)-from the Canada Basin west of Barrow Strait, and from Baffin Bay east thereof. Nearly 25% of nutrients in the central CAA were of Atlantic origin, a substantially larger fraction than surmised previously. Nutrient properties in the CAA were notably not influenced by benthic denitrification. Those in underlying AW in the western CAA, however, were modified by remineralization of Pacific-sourced nutrients, manifested from increases in delta N-15(NO3) relative to the Canada Basin. Properties in the eastern CAA exposed the complex hydrography of Baffin Bay, characterized by lateral intrusions of nutrient-poor winter waters into PWW and of cold mode waters in underlying AW. High delta N-15(NO3) values in AW revealed the presence of a substantial fraction of N from remineralization, stemming from the high productivity in Baffin Bay fueled by Pacific nutrients.

Atmospheric nitrate originates from the oxidation of nitrogen oxides (NOx=NO+NO2) and impacts both tropospheric chemistry and climate. NOx sources, cycling and NOx to nitrate formation pathways are poorly constrained in remote marine regions, especially the Southern Ocean, where pristine conditions serve as a useful proxy for the pre-industrial atmosphere. Here, we measured the isotopic composition (δ15N and δ18O) of atmospheric nitrate in coarse-mode (>1 µm) aerosols collected in the summertime marine boundary layer of the Atlantic Southern Ocean from 34.5 to 70∘ S and across the northern edge of the Weddell Sea. The δ15N–NO3- decreased with latitude from −2.7 ‰ to −42.9 ‰. The decline in δ15N with latitude is attributed to changes in the dominant NOx sources: lightning at the low latitudes, oceanic alkyl nitrates at the mid-latitudes and photolysis of nitrate in snow at the high latitudes. There is no evidence of any influence from anthropogenic NOx sources or equilibrium isotope fractionation. Using air mass back trajectories and an isotope mixing model, we calculate that oceanic alkyl nitrate emissions have a δ15N signature of -21.8±7.6 ‰. Given that measurements of alkyl nitrate contributions to remote nitrogen budgets are scarce, this may be a useful tracer for detecting their contribution in other oceanic regions. The δ18O–NO3- was always less than 70 ‰, indicating that daytime processes involving OH are the dominant NOx oxidation pathway during summer. Unusually low δ18O–NO3- values (less than 31 ‰) were observed at the western edge of the Weddell Sea. The air mass history of these samples indicates extensive interaction with sea-ice-covered ocean, which is known to enhance peroxy radical production. The observed low δ18O–NO3- is therefore attributed to increased exchange of NO with peroxy radicals, which have a low δ18O, relative to ozone, which has a high δ18O. This study reveals that the mid- and high-latitude surface ocean may serve as a more important NOx source than previously thought and that the ice-covered surface ocean impacts the reactive nitrogen budget as well as the oxidative capacity of the marine boundary layer.

Biological dinitrogen fixation is the major source of new nitrogen to marine systems and thus essential to the ocean’s biological pump. Constraining the distribution and global rate of dinitrogen fixation has proven challenging owing largely to uncertainty surrounding the controls thereon. Existing South Atlantic dinitrogen fixation rate estimates vary five-fold, with models attributing most dinitrogen fixation to the western basin. From hydrographic properties and nitrate isotope ratios, we show that the Angola Gyre in the eastern tropical South Atlantic supports the fixation of 1.4–5.4 Tg N.a −1 , 28-108% of the existing (highly uncertain) estimates for the basin. Our observations contradict model diagnoses, revealing a substantial input of newly-fixed nitrogen to the tropical eastern basin and no dinitrogen fixation west of 7.5˚W. We propose that dinitrogen fixation in the South Atlantic occurs in hotspots controlled by the overlapping biogeography of excess phosphorus relative to nitrogen and bioavailable iron from margin sediments. Similar conditions may promote dinitrogen fixation in analogous ocean regions. Our analysis suggests that local iron availability causes the phosphorus-driven coupling of oceanic dinitrogen fixation to nitrogen loss to vary on a regional basis.

Estuaries receive substantial anthropogenic nitrogen loading and are mainly considered net nitrogen sinks. While several studies have identified diverse diazotrophic communities in estuarine sediments, the role of pelagic diazotrophs in these systems is not well understood. We investigated the links between diazotrophic community composition, nitrogenase (nifH) gene expression, N2 fixation, and environmental conditions in Narragansett Bay (USA). Pelagic N2 fixation rates ranged between 0.02 and 9.41 nmol N L−1 d−1 and correlated significantly with fluctuations in diazotroph community composition. These fluctuations were also correlated with temperature, salinity, and mean sea level. The dominant sequences in our pelagic samples were related to sequences previously detected in the bay's sediments and were dominated by nifH gene Clusters I and III. We interpret this as a coupling between sediment and pelagic diazotroph communities and speculate that resuspension plays an important role for pelagic N2 fixation in shallow estuarine environments such as Narragansett Bay. For instance, the finding of active sulfate reducers in the oxygenated water illustrates that the sediment‐pelagic coupling can impact nutrient cycling in shallow environments. The pelagic N2 fixation measured during our study period showed only a minor contribution (< 1%) to the total estimated nitrogen load to Narragansett Bay. However, with intensifying nitrogen management in estuaries, the need to constrain the rates of pelagic N2 fixation in these systems will be essential for estimating nitrogen fluxes within the bay and to the adjacent coastal ecosystems.

Table S1: The starting and ending date, latitude (°S), and longitude(°E) are presented for each aerosol filter deployment.The wind speed (WS; m s⁻¹), atmospheric temperature (Atm T; °C), relative humidity (RH; %), and the number of daylight hours (hrs) were calculated as an average (Avg) over the duration of each filter deployment.For WS, Atm T and RH the standard deviations (SD) are also shown.Filter deployments are separated into early Summer (ES), Weddell Sea (WS) and late Summer (LS) depending on the location and time of sampling. Cruise

Salinity-driven density stratification of the upper Arctic Ocean isolates sea-ice cover and cold, nutrient-poor surface waters from underlying warmer, nutrient-rich waters. Recently, stratification has strengthened in the western Arctic but has weakened in the eastern Arctic; it is unknown if these trends will continue. Here we present foraminifera-bound nitrogen isotopes from Arctic Ocean sediments since 35,000 years ago to reconstruct past changes in nutrient sources and the degree of nutrient consumption in surface waters, the latter reflecting stratification. During the last ice age and early deglaciation, the Arctic was dominated by Atlantic-sourced nitrate and incomplete nitrate consumption, indicating weaker stratification. Starting at 11,000 years ago in the western Arctic, there is a clear isotopic signal of Pacific-sourced nitrate and complete nitrate consumption associated with the flooding of the Bering Strait. These changes reveal that the strong stratification of the western Arctic relies on low-salinity inflow through the Bering Strait. In the central Arctic, nitrate consumption was complete during the early Holocene, then declined after 5,000 years ago as summer insolation decreased. This sequence suggests that precipitation and riverine freshwater fluxes control the stratification of the central Arctic Ocean. Based on these findings, ongoing warming will cause strong stratification to expand into the central Arctic, slowing the nutrient supply to surface waters and thus limiting future phytoplankton productivity. Stratification of the central Arctic Ocean, important for sea-ice persistence, has been controlled by a balance of sea level and freshwater inputs since the last ice age, according to foraminifera-bound nitrogen isotope records that are indicative of surface-water nutrient levels covering the past 35,000 years.

Coastal waters globally are increasingly impacted due to the anthropogenic loading of nitrogen (N) from the watershed. To assess dominant sources contributing to the eutrophication of the Little Narragansett Bay estuary in New England, we carried out an annual study of N loading from the Pawcatuck River. We conducted weekly monitoring of nutrients and nitrate (NO3-) isotope ratios (15N / 14N, 18O / 16O, and 17O / 16O) at the mouth of the river and from the larger of two wastewater treatment facilities (WWTFs) along the estuary, as well as seasonal along-river surveys. Our observations reveal a direct relationship between N loading and the magnitude of river discharge and a consequent seasonality to N loading into the estuary – rendering loading from the WWTFs and from an industrial site more important at lower river flows during warmer months, comprising ∼ 23 % and ∼ 18 % of N loading, respectively. Riverine nutrients derived predominantly from deeper groundwater and the industrial point source upriver in summer and from shallower groundwater and surface flow during colder months – wherein NO3- associated with deeper groundwater had higher 15N / 14N ratios than shallower groundwater. Corresponding NO3- 18O / 16O ratios were lower during the warm season, due to increased biological cycling in-river. Uncycled atmospheric NO3-, detected from its unique mass-independent NO3- 17O / 16O vs. 18O / 16O fractionation, accounted for < 3 % of riverine NO3-, even at elevated discharge. Along-river, NO3- 15N / 14N ratios showed a correspondence to regional land use, increasing from agricultural and forested catchments to the more urbanized watershed downriver. The evolution of 18O / 16O isotope ratios along-river conformed to the notion of nutrient spiraling, reflecting the input of NO3- from the catchment and from in-river nitrification and its coincident removal by biological consumption. These findings stress the importance of considering seasonality of riverine N sources and loading to mitigate eutrophication in receiving estuaries. Our study further advances a conceptual framework that reconciles with the current theory of riverine nutrient cycling, from which to robustly interpret NO3- isotope ratios to constrain cycling and source partitioning in river systems.

Rationale Analyses of the isotope ratios of nitrogen (N-15/N-14) and oxygen (O-18/O-16) in nitrate (NO3-) with the denitrifier method require relatively high sample volumes at low concentrations (<= 1 mu M) to afford sufficient analyte for mass spectrometry, resulting in isotopic offsets compared to more concentrated samples of the same isotopic composition. Methods To uncover the origins of isotopic offsets, we analyzed the N and O isotope ratios of NO3- reference materials spanning concentrations of 0.5-20 mu M. We substantiated the incidence of volume-dependent isotopic offsets, then investigated whether they resulted from (a) incomplete sample recovery during N2O sparging, (b) blanks - bacterial, atmospheric, or in reference material solutions - and (c) oxygen atom exchange with water during the bacterial conversion of NO3- to N2O. Results Larger sample volumes resulted in modest offsets in delta N-15, but substantial offsets in delta O-18. N2O recovery from sparging was less complete at higher volumes, resulting in decreases in delta N-15 and delta O-18 due to associated isotope fractionation. Blanks increased detectably with volume, whereas oxygen atom exchange with water remained constant within batch analyses, being sensitive to neither sample volume nor salinity. The sizeable offsets in delta O-18 with volume are only partially explained by the factors considered in our analysis. Conclusions Our observations argue for bracketing of NO3- samples with reference materials that emulate sample volumes (concentrations) to achieve improved measurement accuracy and foster inter-comparability.