Benthic Denitrification Rates Research Articles

Weak Hypoxia Enhanced Denitrification in a Dissimilatory Nitrate Reduction to Ammonium (DNRA)-Dominated Shallow and Eutrophic Coastal Waterbody, Jinhae Bay, South Korea

The effects of seasonal hypoxia on sediment-water interface nitrogen (N) transformations in Jinhae Bay were examined from 2015 to 2019. The rates of benthic denitrification, anaerobic ammonium oxidation (anammox), dissimilatory nitrate reduction to ammonium (DNRA), nutrient exchange, and sediment oxygen consumption were measured seasonally. The oxygen (O2) and hydrogen sulfide (H2S) depth profiles were measured using microelectrodes. Neither penetration nor consumption of oxygen decreased during hypoxia. Denitrification, anammox, and DNRA ranged from 0 to 0.73, 0.13, and 1.09 mmol N m-2 day-1, respectively. Denitrification, the dominant N removal pathway, increased by 75% while anammox ceased, which led to an overall increase of 55% in the total N2 gas production during hypoxia relative to that during normoxia. Enhanced denitrification is the result of increased coupled nitrification–denitrification due to the intermittent supply of oxygen during bottom water hypoxia (“weak hypoxia”). In the hypoxic period, DNRA decreased by 62%, and the relative contribution of DNRA to the total nitrogen reduction process decreased from 81 to 58%, but it still outperformed denitrification as the main nitrate reduction pathway. Sediments were strong sources of ammonium for the water column, both under normoxia and hypoxia, whereas they were a sink of nitrate from the water column during hypoxia. Bioturbation may be important for maintaining oxygen penetration and consumption in sediments. The dominance of DNRA was mainly due to the relatively high content of sulfide and organic-rich sediments. The repressed macrofaunal activity and increased coupling of nitrification and denitrification during hypoxia may have contributed to enhanced denitrification. Taken together, the overall dominance of DNRA might contribute to the development and maintenance of eutrophication and seasonal hypoxia in this system. However, in contrast to the previous results, denitrification was enhanced during “weak hypoxia,” which might be helpful in alleviating eutrophication.

High Incidence Hypoxia Increases Benthic Nitrogen Retention but Decreases Nitrogen Removal in Seasonally Hypoxic Areas off the Changjiang Estuary

AbstractHow the partitioning between dissimilatory nitrate reduction to ammonium (DNRA) and N2 production responds to hypoxia in seasonally hypoxic estuaries has not been well understood. In this study, 15N tracer incubations were used to investigate benthic denitrification, anammox and DNRA rates for three years (2016–2018) across seasonally hypoxic areas off the Changjiang Estuary. Dissolved oxygen contents in bottom water were significantly lower in July than in February/March. Concentrations of NH4+ and NOx– in bottom water throughout the sampling sites did not vary between seasons. Rates of benthic denitrification (0.45–6.55 nmol N g−1 h−1), anammox (0.02–0.30 nmol N g−1 h−1) and DNRA (0.06–0.59 nmol N g−1 h−1) varied largely along the estuary. Denitrification, anammox and DNRA rates did not vary significantly between inshore and offshore sites. However, denitrification rates were significantly lower and DNRA rates were significantly higher in consistently than transiently hypoxic sites. Anammox rates were slightly lower in consistently than transiently hypoxic sites. Ratios of DNRA to N2 production were in a range of 0.015–0.53, and were significantly higher in consistently than transiently hypoxic sites, but did not vary significantly between inshore and offshore sites. The lower temperature sensitivity of denitrification and anammox compared to their Q10 values further indicated that hypoxia may inhibit N2 production. These results suggest that high incidence hypoxia can decrease denitrification and anammox rates but increase DNRA rates with important implications on N retention versus removal, thereby exacerbating eutrophication and hypoxia in estuarine and coastal environments.

Seasonal cycle of benthic denitrification and DNRA in the aphotic coastal zone, northern Baltic Sea

Current knowledge on the seasonality of benthic nitrate reduction pathways in the aphotic, density stratified coastal zone of the Baltic Sea is largely based on data from muddy sediments, neglecting the potential contribution of sandy sediments. To gain a more comprehensive understanding of seasonality in this part of the Baltic Sea coast, we measured rates of benthic denitrification, anammox and dissimilatory nitrate reduction to ammonium (DNRA) monthly in the ice-free period of 2016 in both sandy and muddy aphotic sediments, northwestern Gulf of Finland. No anammox was observed. The seasonal cycle of denitrification in both sediment types was related to the hydrography-driven development of bottom water temperature. The seasonal cycle of DNRA was less clear and likely connected to a combination of bottom water temperature, carbon to nitrogen ratio, and substrate competition with denitrification. Denitrification and DNRA rates were 50-80 and 20% lower in the sandy than in the muddy sediment. The share of DNRA in total nitrate reduction, however, was higher in the sandy than in the muddy sediment, being (by ~50%) the highest DNRA share in sandy sediments so far measured. Our data add to the small pool of published studies showing significant DNRA in both cold and/or sandy sediments and suggest that DNRA is currently underestimated in the Baltic coastal nitrogen filter. Our results furthermore emphasize that the various environmental conditions of a coastal habitat (light regime, hydrography, and geomorphology) affect biogeochemical element cycling and thus need to be considered in data interpretation.

Comparison of methods for determining biogeochemical fluxes from a restored oyster reef.

Oyster reef restoration can significantly increase benthic denitrification rates. Methods applied to measure nutrient fluxes and denitrification from oyster reefs in previous studies include incubations of sediment cores collected adjacent to oyster clumps, benthic chambers filled with intact reef segments that have undergone in situ equilibration and ex situ incubation, and cores with single oysters. However, fluxes of nutrients vary by orders of magnitude among oyster reefs and methods. This study compares two methods of measuring nutrient and metabolic fluxes on restored oyster reefs: incubations including intact segments of oyster reef and incubations containing oyster clumps without underlying sediments. Fluxes of oxygen (O2), dissolved inorganic carbon (DIC), ammonium (NH4+), combined nitrate and nitrite (NO2/3-), di-nitrogen (N2), and soluble reactive phosphorus (SRP) were determined in June and August in Harris Creek, a tributary of the Chesapeake Bay, Maryland, USA. Regression of fluxes measured from clumps alone against those measured from intact reef segments showed significant positive relationships for O2, DIC, NH4+, and SRP (R2 = 0.920, 0.61, 0.26, and 0.52, respectively). Regression of clump fluxes against the oyster tissue biomass indicates significant positive relationships for O2 and NH4+, marginally significant and positive relationships for DIC and N2, and no significant relationship for NO2/3- or SRP. Although these results demonstrate that the incubation of oyster clumps without underlying sediments does not accurately represent biogeochemical fluxes measured from the whole oyster and sediment community, this work supports the need to understand the balance between the metabolism of oysters and local sediments to correctly estimate biogeochemical rates.

Do wetlands enhance downstream denitrification in agricultural landscapes?

AbstractWetlands are known to be effective sinks for nitrate. Wetland restoration and construction have gained traction as viable conservation measures to improve water quality in intensively managed agricultural landscapes. In addition to reducing nitrate in situ, wetlands may have impacts on water chemistry and temperature dynamics that extend beyond the confines of the wetlands themselves. Nonsaturating nitrate concentrations (NO3−), enhanced organic carbon effluxes, and altered temperature dynamics in streams downstream of wetlands could all affect denitrification rates within a stream network, potentially extending water quality benefits beyond wetland boundaries. We investigated the effect of wetlands on water chemistry, water temperature, and benthic denitrification rates in downstream agricultural ditches through a field measurement campaign over the open water season. We found that although ditches located downstream of wetlands had lower NO3− and higher DOC, ditch denitrification rate was not significantly altered by the presence of upstream wetlands. Rather, wetlands indirectly affected denitrification within ditches by strongly influencing the stoichiometry of the two limiting resources, NO3− and organic carbon. Peak denitrification rates in ditches were observed when DOC and NO3− supplies were approximately balanced, that is, at DOC: NO3− ratios that were near the microbial requirement for denitrification. NO3− limitation occurred primarily at sites with >3% wetland cover, and in the fall season at all sites, and DOC limitation occurred primarily at sites with <1% wetland cover. Temperature was found to be a secondary control that was important only when NO3− and DOC availabilities were balanced. Our results suggest that wetland restoration and construction targeting nitrate reduction within intensively agriculturally managed basins should be implemented in a way that promotes balanced resource availability throughout fluvial networks. Wetlands are an important regulator of resource availability and thus could be used to create conditions that maximize denitrification in NO3−‐enriched watersheds.

Spatial variation in rates of benthic denitrification and environmental controls in four New Zealand estuaries

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 556:59-77 (2016) - DOI: https://doi.org/10.3354/meps11865 Spatial variation in rates of benthic denitrification and environmental controls in four New Zealand estuaries Catherine Gongol1, Candida Savage1,2,* 1Department of Marine Science, University of Otago, PO Box 56, Dunedin, New Zealand 2Department of Biological Sciences, University of Cape Town, Rondebosch, South Africa *Corresponding author: candida.savage@otago.ac.nz ABSTRACT: Denitrification plays a key role in global biogeochemical cycles as the main nitrogen (N) loss process that limits availability of N to primary producers. This study presents the first measurements of sediment denitrification rates for New Zealand estuaries. Spatially explicit rates of direct denitrification (Dw) and coupled nitrification-denitrification (Dn) were quantified using the isotope pairing method in 4 oligotrophic to moderately nutrient-enriched estuaries on the South Island, New Zealand (Avon-Heathcote, Waikouaiti, Tokomairiro, Tautuku). Coupled nitrification-denitrification dominated (>86%), with most sites having low dissolved inorganic nitrogen (DIN) concentrations (<30 µmol l-1). Rates of total denitrification (Dtot) and Dn were negatively correlated with percent sand and positively related to oxygen penetration depth (OPD). Surface sediment chlorophyll a concentrations and percent carbon (%C) were also identified as significant predictors of Dtot and Dn in the single-factor models. By contrast, Dw was the main nitrogen removal process at moderately nutrient-enriched sites and was positively related to DIN concentrations in the water column and negatively correlated with percent sand. OPD and %C were also identified as significant predictors of Dw. While Dw was positively related to DIN concentrations, overall rates of denitrification were not, demonstrating that predictive relationships between N loading and removal are not always possible in oligotrophic estuaries. Our findings demonstrate the importance of grain size as a driver of denitrification rates in estuaries and imply that an increase in fine-grained sediments from land-use intensification may alter rates of N removal in estuaries through complex direct and indirect processes. KEY WORDS: Denitrification · Estuaries · New Zealand · Nitrogen · Benthic · Sediment grain size · Isotope pairing method · Oxygen flux Full text in pdf format PreviousNextCite this article as: Gongol C, Savage C, (2016) Spatial variation in rates of benthic denitrification and environmental controls in four New Zealand estuaries. Mar Ecol Prog Ser 556:59-77. https://doi.org/10.3354/meps11865 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 556. Online publication date: September 08, 2016 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2016 Inter-Research.

Chronic exposure of river sediments to environmentally relevant levels of tetracycline affects bacterial communities but not denitrification rates

The effects of tetracycline (TC) at chronic sub-inhibitory exposure concentrations on benthic denitrification rates and bacterial communities were explored. River sediments were continuously exposed to different TC concentrations (0.5, 20 and 10,000 μg L(-1)) for 2 weeks in flow-through reactors allowing denitrification and bacterial growth conditions. Bacterial communities were fingerprinted by Denaturing Gradient Gel Electrophoresis of 16S rRNA gene amplification products. Cultivable denitrifiers enriched from the sediment were tested for TC resistance (2-128 mg L(-1)). Denitrification rates were unaffected by exposure to TC, regardless of concentration. In contrast, the bacterial community composition changed significantly from sub-inhibitory (ng-μg L(-1)) to therapeutic (mg L(-1)) exposure concentrations. Furthermore the cultivable denitrifiers showed a high TC sensitivity (<4 mg L(-1)). Maintenance of efficient benthic denitrification rates, even at the highest level of TC exposure most likely originated from an adaptation of the autochthonous bacterial community where dominant species become those that acquire, or already have resistance to antibiotics.

Denitrification in a large river: consideration of geomorphic controls on microbial activity and community structure

Ecological theory argues that the controls over ecosystem processes are structured hierarchically, with broader-scale drivers acting as constraints over the interactions and dynamics at nested levels of organization. In river ecosystems, these interactions may arise from broadscale variation in channel form that directly shapes benthic habitat structure and indirectly constrains resource supply and biological activity within individual reaches. To evaluate these interactions, we identified sediment characteristics, water chemistry, and denitrifier community structure as factors influencing benthic denitrification rates in a sixth-order river that flows through two physiographic provinces and the transitional zone between them, each with distinct geomorphological properties. We found that denitrification rates tracked spatial changes in sediment characteristics and varied seasonally with expected trends in stream primary production. Highest rates were observed during the spring and summer seasons in the physiographic province dominated by fine-grained sediments, illustrating how large-scale changes in river structure can constrain the location of denitrification hotspots. In addition, nirS and nirK community structure each responded differently to variation in channel form, possibly due to changes in dissolved oxygen and organic matter supply. This shift in denitrifier community structure coincident with higher rates of N removal via denitrification suggests that microbial community structure may influence biogeochemical processes.

The role of benthic foraminifera in the benthic nitrogen cycle of the Peruvian oxygen minimum zone

Abstract. The discovery that foraminifera are able to use nitrate instead of oxygen as an electron acceptor for respiration has challenged our understanding of nitrogen cycling in the ocean. It was thought before that only prokaryotes and some fungi are able to denitrify. Rate estimates of foraminiferal denitrification have been very sparse and limited to specific regions in the oceans, not comparing stations along a transect of a certain region. Here, we present estimates of benthic foraminiferal denitrification rates from six stations at intermediate water depths in and below the Peruvian oxygen minimum zone (OMZ). Foraminiferal denitrification rates were calculated from abundance and assemblage composition of the total living fauna in both surface and subsurface sediments, as well as from individual species specific denitrification rates. A comparison with total benthic denitrification rates as inferred by biogeochemical models revealed that benthic foraminifera probably account for the total denitrification in shelf sediments between 80 and 250 m water depth. The estimations also imply that foraminifera are still important denitrifiers in the centre of the OMZ around 320 m (29–50% of the benthic denitrification), but play only a minor role at the lower OMZ boundary and below the OMZ between 465 and 700 m (2–6% of total benthic denitrification). Furthermore, foraminiferal denitrification has been compared to the total benthic nitrate loss measured during benthic chamber experiments. The estimated foraminiferal denitrification rates contribute 2 to 46% to the total nitrate loss across a depth transect from 80 to 700 m, respectively. Flux rate estimates range from 0.01 to 1.3 mmol m−2 d−1. Furthermore we show that the amount of nitrate stored in living benthic foraminifera (3 to 3955 μmol L−1) can be higher by three orders of magnitude as compared to the ambient pore waters in near-surface sediments sustaining an important nitrate reservoir in Peruvian OMZ sediments. The substantial contribution of foraminiferal nitrate respiration to total benthic nitrate loss at the Peruvian margin, which is one of the main nitrate sink regions in the world ocean, underpins the importance of the previously underestimated role of benthic foraminifera in global biogeochemical cycles.

Benthic metabolism and nitrogen dynamics in an urbanised tidal creek: Domination of DNRA over denitrification as a nitrate reduction pathway

Benthic oxygen and nutrient fluxes and nitrate reduction rates were determined seasonally under light and dark conditions at three sites in a micro-tidal creek within an urbanised catchment (Saltwater Creek, Australia). It was hypothesized that stormwater inputs of organic matter and inorganic nitrogen would stimulate rates of benthic metabolism and nutrient recycling and preferentially stimulate dissimilatory nitrate reduction to ammonium (DNRA) over denitrification as a pathway for nitrate reduction. Stormwaters greatly influenced water column dissolved inorganic nitrogen (DIN) and suspended solids concentrations with values following a large rainfall event being 5–20-fold greater than during the preceding dry period. Seasonally, maximum and minimum water column total dissolved nitrogen (TDN) and DIN concentrations occurred in the summer (wet) and winter (dry) seasons. Creek sediments were highly heterotrophic throughout the year, and strong sinks for oxygen, and large sources of dissolved organic and inorganic nitrogen during both light and dark incubations, although micro-phytobenthos (MPB) significantly decreased oxygen consumption and N-effluxes during light incubations due to photosynthetic oxygen production and photoassimilation of nutrients. Benthic denitrification rates ranged from 3.5 to 17.7 μmol N m2 h−1, denitrification efficiencies were low (<1–15%) and denitrification was a minor process compared to DNRA, which accounted for ∼75% of total nitrate reduction.Overall, due to the low denitrification efficiencies and high rates of N-regeneration, Saltwater Creek sediments would tend to increase rather than reduce dissolved nutrient loads to the downstream Gold Coast Broadwater and Moreton Bay systems. This may be especially true during wet periods when increased inputs of particulate organic nitrogen (PON) and suspended solids could respectively enhance rates of N-regeneration and decrease light availability to MPB, reducing their capacity to ameliorate N-effluxes through photoassimilation.

Changes in Benthic Denitrification, Nitrate Ammonification, and Anammox Process Rates and Nitrate and Nitrite Reductase Gene Abundances along an Estuarine Nutrient Gradient (the Colne Estuary, United Kingdom)

Estuarine sediments are the location for significant bacterial removal of anthropogenically derived inorganic nitrogen, in particular nitrate, from the aquatic environment. In this study, rates of benthic denitrification (DN), dissimilatory nitrate reduction to ammonium (DNRA), and anammox (AN) at three sites along a nitrate concentration gradient in the Colne estuary, United Kingdom, were determined, and the numbers of functional genes (narG, napA, nirS, and nrfA) and corresponding transcripts encoding enzymes mediating nitrate reduction were determined by reverse transcription-quantitative PCR. In situ rates of DN and DNRA decreased toward the estuary mouth, with the findings from slurry experiments suggesting that the potential for DNRA increased while the DN potential decreased as nitrate concentrations declined. AN was detected only at the estuary head, accounting for approximately 30% of N2 formation, with 16S rRNA genes from anammox-related bacteria also detected only at this site. Numbers of narG genes declined along the estuary, while napA gene numbers were stable, suggesting that NAP-mediated nitrate reduction remained important at low nitrate concentrations. nirS gene numbers (as indicators of DN) also decreased along the estuary, whereas nrfA (an indicator for DNRA) was detected only at the two uppermost sites. Similarly, nitrate and nitrite reductase gene transcripts were detected only at the top two sites. A regression analysis of log(n + 1) process rate data and log(n + 1) mean gene abundances showed significant relationships between DN and nirS and between DNRA and nrfA. Although these log-log relationships indicate an underlying relationship between the genetic potential for nitrate reduction and the corresponding process activity, fine-scale environmentally induced changes in rates of nitrate reduction are likely to be controlled at cellular and protein levels.

DENITRIFICATION IN NITRATE-RICH STREAMS: APPLICATION OF N2:Ar AND15N-TRACER METHODS IN INTACT CORES

Rates of benthic denitrification were measured using two techniques, membrane inlet mass spectrometry (MIMS) and isotope ratio mass spectrometry (IRMS), applied to sediment cores from two NO3(-)-rich streams draining agricultural land in the upper Mississippi River Basin. Denitrification was estimated simultaneously from measurements of N2:Ar (MIMS) and 15N[N2] (IRMS) after the addition of low-level 15NO3- tracer (15N:N = 0.03-0.08) in stream water overlying intact sediment cores. Denitrification rates ranged from about 0 to 4400 micromol N x m(-2) x h(-1) in Sugar Creek and from 0 to 1300 micromol N x m(-2) x h(-1) in Iroquois River, the latter of which possesses greater streamflow discharge and a more homogeneous streambed and water column. Within the uncertainties of the two techniques, there is good agreement between the MIMS and IRMS results, which indicates that the production of N2 by the coupled process of nitrification/denitrification was relatively unimportant and surface-water NO3- was the dominant source of NO3- for benthic denitrification in these streams. Variation in stream NO3- concentration (from about 20 micromol/L during low discharge to 1000 micromol/L during high discharge) was a significant control of benthic denitrification rates, judging from the more abundant MIMS data. The interpretation that NO3- concentration directly affects denitrification rate was corroborated by increased rates of denitrification in cores amended with NO3-. Denitrification in Sugar Creek removed < or = 11% per day of the instream NO3- in late spring and removed roughly 15-20% in late summer. The fraction of NO3- removed in Iroquois River was less than that of Sugar Creek. Although benthic denitrification rates were relatively high during periods of high stream flow, when NO3 concentrations were also high, the increase in benthic denitrification could not compensate for the much larger increase in stream NO3- fluxes during high flow. Consequently, fractional NO3- losses were relatively low during high flow.

Denitrification in marine sediments: A model study

The rate and factors controlling denitrification in marine sediments have been investigated using a prognostic diagenetic model. The model is forced with observed carbon fluxes, bioturbation and sedimentation rates, and bottom water conditions. It can reproduce rates of aerobic mineralization, denitrification, and fluxes of oxygen, nitrate, and ammonium. The globally integrated rate of denitrification is estimated by this model to be about 230–285 Tg N yr−1, with about 100 Tg N yr−1 occurring in shelf sediments. This estimate is significantly higher than literature estimates (12–89 Tg N yr−1), mainly because of a proposed upward revision of denitrification rates in slope and deep‐sea sediments. Higher sedimentary denitrification estimates require a revision of the marine nitrogen budget and lowering of the oceanic residence time of nitrogen down to about 2×103 years and are consistent with reported low N/P remineralization ratios between 1000 and 3000 m. Rates of benthic denitrification are most sensitive to the flux of labile organic carbon arriving at the sediment‐water interface and bottom water concentrations of nitrate and oxygen. Denitrification always increases when bottom water nitrate increases but may increase or decrease if oxygen in the bottom water increases. Nitrification is by far the most important source of nitrate for denitrification, except for organic‐rich sediments underlying oxygen‐poor and nitrate‐rich water.

Denitrification, nutrient regeneration and carbon mineralization in sediments of Galveston Bay, Texas, USA

Rates of benthic denitrification, oxygen consumption and nutrient regeneration were measured during winter, spring and summer in Galveston Bay (Texas, USA) sediments. Denitrification ranged from 0 to 47 pm01 N2 m-2 h' with maximal rates generally occurring in the summer and the upper estuary. Oxygen consumption rates ranged from 38 pm01 O2 m-2 h' in the winter to 353 pm01 O2 m-' h-' in the summer and were correlated with denitrification rates. Variations in bay water temperature accounted for 52 % of the variability associated with denitrification rates whereas only 28% of the variability could be attributed to organic carbon content and 15% to salinity, indicating a predominance of temporal over spatial factors in controlling estuarine rates of denitrification. In the spring and summer, denitrification was responsible for the majority (73 and 80%, respectively) of the total benthic inorganic nitrogen efflux while in the winter, nitrogen fluxes were dominated (80 %) by ammonium. At salinities less than 6 L , cation exchange interactions may have played an important role in retaining ammonium in the sediment, producing the higher rates of denitrification found in the upper estuary. Dissolved inorganic carbon flux was used as a measure of total organic carbon mineralization. The average molar C:N of the remineralized substrate (5.2) was lower than the average C:N of the sediments (12.6) indicating preferent~al remineralization of nitrogen relative to carbon. Molar C:O ratlos suggested that anaerobic carbon m~neralization and the storage of its reduced end-products is more prevalent in the lower estuary and In the winter. Denitrifiers were responsible for 37 and 13 % of the total benthic carbon mineralization in the upper and lower estuary, respectively. Denitrification appears to be a greater contributor to total carbon mineralization than previously considered. Nearly one-third of the total sediment oxygen consumption was attributed to nitrification. Galveston Bay sediment denitrification and oxygen consumption rates and nutrient fluxes were lower but comparable to those of other Gulf of Mexico estuaries. Differences among the estuaries examined are attributed mainly to sediment organic matter content.

Eutrophication and the rate of denitrification and N20 production in coastal marine sediments

Large (13 m3, 5 m deep) microcosms with coupled pelagic and benthic components were used to measure the effect of nutrient loading and eutrophication in coastal marine ecosystems on the rates of benthic denitrification (N2) and N2O production. After 3 months of daily nutrient addition, average denitrification rates ranged from about 300 µmol N m−2 h−1 in the sediments of the control microcosm to 880 in the most enriched microcosm. which received 65 times the nutrient input of the control. Increases in the production of N2O were more dramatic and increased by a factor of about 100, from 0.56 µmol N m−2 h−1 in the control to 51 in the most enriched microcosm. Although there was a clear increase in the denitrification rate in the more eutrophic systems, the amount of fixed nitrogen removed was a constant or progressively smaller fraction of the nitrogen input. Even in the most enriched microcosm, at least 16% of the N input was removed by denitrification.