Crown-of-thorns starfish complete their larval phase eating only nitrogen-fixing Trichodesmium cyanobacteria.

Cyanobacteria of the genus Trichodesmium form extensive blooms that supply new N to nutrient-poor marine ecosystems. Yet little is known about what eats Trichodesmium. In this laboratory study, we show that one of the greatest threats to coral reefs, predatory crown-of-thorns starfish (CoTS), Acanthaster sp., completes their larval phase feeding solely on Trichodesmium. We observed Trichodesmium erythraeum CMP1985 in the stomachs of larvae using florescence microscopy and traced the assimilation of nitrogen from labeled trichomes into larval tissues using stable isotopes. Some larvae fed T. erythraeum were morphologically ready to become benthic juveniles after 19 days. Given that Trichodesmium can be food for CoTS, reported increases in Trichodesmium could be a driving factor in the heightened frequency of CoTS population irruptions that have devastated coral reefs in past decades. Future studies could test this through investigating the diets of wild larvae and incorporating Trichodesmium abundance into models of CoTS population dynamics.

Characterization of the redox-active extracellular polymeric substances in an anaerobic methanotrophic consortium.

Anaerobic oxidation of methane (AOM) is a microbial process of importance in the global carbon cycle. AOM is predominantly mediated by anaerobic methanotrophic archaea (ANME), the physiology of which is still poorly understood. Here we present a new addition to the current physiological understanding of ANME by examining, for the first time, the biochemical and redox-active properties of the extracellular polymeric substances (EPS) of an ANME enrichment culture. Using a 'Candidatus Methanoperedens nitroreducens'-dominated methanotrophic consortium as the representative, we found it can produce an EPS matrix featuring a high protein-to-polysaccharide ratio of ∼8. Characterization of EPS using FTIR revealed the dominance of protein-associated amide I and amide II bands in the EPS. XPS characterization revealed the functional group of C-(O/N) from proteins accounted for 63.7% of total carbon. Heme-reactive staining and spectroscopic characterization confirmed the distribution of c-type cytochromes in this protein-dominated EPS, which potentially enabled its electroactive characteristic. Redox-active c-type cytochromes in EPS mediated the EET of 'Ca. M. nitroreducens' for the reduction of Ag+ to metallic Ag, which was confirmed by both ex-situ experiments with extracted soluble EPS and in-situ experiments with pristine EPS matrix surrounding cells. The formation of nanoparticles in the EPS matrix during in-situ extracellular Ag + reduction resulted in a relatively lower intracellular Ag distribution fraction, beneficial for alleviating the Ag toxicity to cells. The results of this study provide the first biochemical information on EPS of anaerobic methanotrophic consortia and a new insight into its physiological role in AOM process.

Nitrate-driven anaerobic oxidation of ethane and butane by bacteria.

The short-chain gaseous alkanes (ethane, propane, and butane; SCGAs) are important components of natural gas, yet their fate in environmental systems is poorly understood. Microbially mediated anaerobic oxidation of SCGAs coupled to nitrate reduction has been demonstrated for propane, but is yet to be shown for ethane or butane-despite being energetically feasible. Here we report two independent bacterial enrichments performing anaerobic ethane and butane oxidation, respectively, coupled to nitrate reduction to dinitrogen gas and ammonium. Isotopic 13C- and 15N-labelling experiments, mass and electron balance tests, and metabolite and meta-omics analyses collectively reveal that the recently described propane-oxidizing "Candidatus Alkanivorans nitratireducens" was also responsible for nitrate-dependent anaerobic oxidation of the SCGAs in both these enrichments. The complete genome of this species encodes alkylsuccinate synthase genes for the activation of ethane/butane via fumarate addition. Further substrate range tests confirm that "Ca. A. nitratireducens" is metabolically versatile, being able to degrade ethane, propane, and butane under anoxic conditions. Moreover, our study proves nitrate as an additional electron sink for ethane and butane in anaerobic environments, and for the first time demonstrates the use of the fumarate addition pathway in anaerobic ethane oxidation. These findings contribute to our understanding of microbial metabolism of SCGAs in anaerobic environments.

Decoupling the simultaneous effects of NO2-, pH and free nitrous acid on N2O and NO production from enriched nitrifying activated sludge.

In the pursuit of energy and carbon neutrality, nitrogen removal technologies have been developed featuring nitrite (NO2-) accumulation. However, high NO2- accumulations are often associated with stimulated greenhouse gas (i.e., nitrous oxide, N2O) emissions. Furthermore, the coexistence of free nitrous acid (FNA) formed by NO2- and proton (pH) makes the consequence of NO2- accumulation on N2O emissions complicated. The concurrent three factors, NO2-, pH and FNA may play different roles on N2O and nitric oxide (NO) emissions simultaneously, which has not been systematically studied. This study aims to decouple the effects of NO2- (0-200 mg N/L), pH (6.5-8) and FNA (0-0.15 mg N/L) on the N2O and NO production rates and the production pathways by ammonia oxidizing bacteria (AOB), with the use of a series of precisely executed batch tests and isotope site-preference analysis. Results suggested the dominant factors affecting the N2O production rate were NO2- and FNA concentrations, while pH alone played a relatively insignificant role. The most influential factor shifted from NO2- to FNA as FNA concentrations increased from 0 to 0.15 mg N/L. At concentrations below 0.0045 mg HNO2-N/L, nitrite rather than FNA played a significant role stimulating N2O production at elevated nitrite concentrations. The inhibition effect of FNA emerged with further increase of FNA between 0.0045-0.015 mg HNO2-N/L, weakening the promoting effect of increased nitrite. While at concentrations above 0.015 mg HNO2-N/L, FNA inhibited N2O production especially from nitrifier denitrification pathway with the level of inhibition linearly correlated with the FNA concentration. pH and the nitrite concentration regulated the production pathways, with elevated pH promoting the nitrifier nitrification pathway, while elevated NO2- concentrations promoting the nitrifier denitrification pathway. In contrast to N2O, NO emission was less susceptible to FNA at concentrations up to 0.015 mg N/L but was stimulated by increasing NO2- concentrations. This study, for the first time, distinguished the effects of pH, NO2- and FNA on N2O and NO production, thereby providing support to the design and operation of novel nitrogen removal systems with NO2- accumulation.

Stable isotopes of nitrate (δ15N and δ18O) as functional indicators of nitrogen processing in constructed wetlands.

The effectiveness of nitrogen removal in wetlands relies heavily on the biological processes that control its removal. Here, we used δ15N and δ18O of nitrate (NO3-) to assess the presence and the dominance of transformation processes of nitrogen in two urban water treatment wetlands in Victoria, Australia over two rainfall events. Laboratory incubation experiments were undertaken in both light and dark to measure the isotopic fractionation factor of nitrogen assimilation (by periphyton and algae) and benthic denitrification (using bare sediment). Highest isotopic fractionations were observed for nitrogen assimilation by algae and periphyton in the light, 15ε = -14.6 to -25 ‰ while the 15ε = -1.5 ‰ in bare sediment, consistent with that of benthic denitrification. Transect water samplings of the wetlands showed different rainfall patterns (discrete versus continuous) affect the removal capability of the wetlands. During the discrete event sampling, the observed 15ε of NO3- (an average of 3.0 to 4.3 ‰) within the wetland falls between the experimental 15ε of benthic denitrification and assimilation; coinciding with the decrease in NO3- concentrations, suggesting that both denitrification and assimilation were important removal pathways. Depletion of δ15N-NO3- throughout the whole wetland system also suggested the influence of water column nitrification during this time. In contrast, during continuous rain events, no fractionation effect was observed within the wetland and was consistent with limited NO3- removal. The difference in fractionation factors within the wetland during different sampling conditions suggested that nitrate removal was highly likely limited by changes in overall nutrient inputs, residence time and water temperature which impeded biological uptake or removal. These highlight that consideration of sampling condition is crucial when assessing the efficacy of a wetland in removing nitrogen.

Anaerobic oxidation of propane coupled to nitrate reduction by a lineage within the class Symbiobacteriia.

Anaerobic microorganisms are thought to play a critical role in regulating the flux of short-chain gaseous alkanes (SCGAs; including ethane, propane and butane) from terrestrial and aquatic ecosystems to the atmosphere. Sulfate has been confirmed to act as electron acceptor supporting microbial anaerobic oxidation of SCGAs, yet several other energetically more favourable acceptors co-exist with these gases in anaerobic environments. Here, we show that a bioreactor seeded with biomass from a wastewater treatment facility can perform anaerobic propane oxidation coupled to nitrate reduction to dinitrogen gas and ammonium. The bioreactor was operated for more than 1000 days, and we used 13C- and 15N-labelling experiments, metagenomic, metatranscriptomic, metaproteomic and metabolite analyses to characterize the microbial community and the metabolic processes. The data collectively suggest that a species representing a novel order within the bacterial class Symbiobacteriia is responsible for the observed nitrate-dependent propane oxidation. The closed genome of this organism, which we designate as 'Candidatus Alkanivorans nitratireducens', encodes pathways for oxidation of propane to CO2 via fumarate addition, and for nitrate reduction, with all the key genes expressed during nitrate-dependent propane oxidation. Our results suggest that nitrate is a relevant electron sink for SCGA oxidation in anaerobic environments, constituting a new microbially-mediated link between the carbon and nitrogen cycles.

Crop fertilisation potential of phosphorus in hydrochars produced from sewage sludge.

Sewage sludges are a rich underused source of phosphorus (P) which contributes to environmental degradation, yet if recaptured, could return significant amounts of P to agricultural systems. Hydrothermal carbonisation (HTC) can efficiently recover P, with the added ability to transform P species into potentially more desirable forms for direct application to crops. P dynamics in hydrochars have primarily examined P speciation and chemical extractability as indicators of P bioavailability, but few studies directly evaluate the agronomic effectiveness of hydrochars as P fertilisers. As such, there is a clear need to assess the suitability of hydrochar as a source of bioavailable P in plant systems and the influence of HTC synthesis conditions. Response Surface Modelling of HTC synthesis conditions (pH, temperature and time), revealed initial pH significantly influence P distribution. Mild conditions of 180 °C for 30 min at pH 8.0 maximised P recovery (99%) along with carbon (62%) and nitrogen (43%) in hydrochars. Systematic characterisation of hydrochar P by chemical extraction and P L2,3-edge X-ray absorption near edge spectroscopy revealed H2O, NaHCO3 and NaOH- P fractions were significantly (p < 0.05) reduced in all hydrochars, while HCl-P fraction increased with HTC temperatures at pH 7. In contrast, P L2,3-edge XANES spectra were remarkably similar in raw sludges and corresponding hydrochars, regardless of HTC temperature or pH, revealing P was predominantly present as ferric phosphate with some hydroxyapatite. Multiple linear regression modelling suggested a significant relationship between chemical extractability and P bioavailability to wheat present in the raw sludges and hydrochars. This research provides further insight into the potential to use hydrothermal treatment for recovery and agricultural reuse of P, the importance of operational conditions on P transformation and the relationship between P speciation and bioavailability. The value of sewage sludge in a more sustainable global P cycle is also highlighted.

Microbially facilitated nitrogen cycling in tropical corals.

Tropical scleractinian corals support a diverse assemblage of microbial symbionts. This 'microbiome' possesses the requisite functional diversity to conduct a range of nitrogen (N) transformations including denitrification, nitrification, nitrogen fixation and dissimilatory nitrate reduction to ammonium (DNRA). Very little direct evidence has been presented to date verifying that these processes are active within tropical corals. Here we use a combination of stable isotope techniques, nutrient uptake calculations and captured metagenomics to quantify rates of nitrogen cycling processes in a selection of tropical scleractinian corals. Denitrification activity was detected in all species, albeit with very low rates, signifying limited importance in holobiont N removal. Relatively greater nitrogen fixation activity confirms that corals are net N importers to reef systems. Low net nitrification activity suggests limited N regeneration capacity; however substantial gross nitrification activity may be concealed through nitrate consumption. Based on nrfA gene abundance and measured inorganic N fluxes, we calculated significant DNRA activity in the studied corals, which has important implications for coral reef N cycling and warrants more targeted investigation. Through the quantification and characterisation of all relevant N-cycling processes, this study provides clarity on the subject of tropical coral-associated biogeochemical N-cycling.

The legacy and drivers of groundwater nutrients and pesticides in an agriculturally impacted Quaternary aquifer system.

Nutrient and pesticide pollution are among the major threats to groundwater quality in agriculturally impacted aquifers. Understanding their legacy effects and drivers are important to protect aquifers from exposures to contamination. However, the complexities of groundwater flowpaths make it difficult to predict the time-scales of groundwater flow and contaminant transport. To determine these controls of groundwater nutrient and pesticides in an aquifer system underlying an intensive agricultural area in the Great Barrier Reef catchment, Australia, we sampled tritium (3H) to estimate groundwater-age, nutrient and pesticide concentrations to investigate groundwater contamination, and nitrogen (ẟ15N-NO3-) and oxygen (ẟ18O-NO3-) isotopes to determine groundwater nitrate dynamics. We, then, constructed high-resolution 3D geological and groundwater flow models of the aquifer system to determine the role of the geologic heterogeneity on the observed nutrient and pesticide concentrations. Groundwater 3H derived ages, and nutrient and pesticide concentrations did not follow distinct spatial trends. ẟ15N-NO3- and ẟ18O-NO3- values indicated that nitrification and denitrification processes influenced nitrate dynamics in the aquifer system; however, they were not solely able to explain the entire 3D variability. The 3D geologic modelling identified possible preferential flowpaths and perched systems, which helped to explain the observed groundwater-age, nutrient and pesticide variabilities. Old-groundwater (~100-years) was found in shallow depths (<15 m) where perched systems were identified. In areas with preferential flowpaths, young-groundwater (⁓1-year) with significant nitrate (~12 mg-N/L) and pesticides (up to 315 ng/L) concentrations were detected at deeper depths (>25 m), below perched and locally confined systems. Downward increasing groundwater-age, and decreasing nutrient and pesticide concentrations were detected in the unconfined aquifer, while old-groundwater (~160-years) and lower nitrate (<3 mg-N/L) and pesticides (<2 ng/L) concentrations were detected in the confined systems. This study demonstrates the importance of understanding both the geology and the hydrogeology of an area before deploying monitoring studies and/or making conclusions from tritium, nutrient and pesticide data alone.

Coral skeletons reveal the history of nitrogen cycling in the coastal Great Barrier Reef.

Anthropogenic nutrient discharge to coastal marine environments is commonly associated with excessive algal growth and ecosystem degradation. However in the world's largest coral reef ecosystem, the Great Barrier Reef (GBR), the response to enhanced terrestrial nutrient inputs since European settlement in the 1850's remains unclear. Here we use a 333 year old composite record (1680-2012) of 15N/14N in coral skeleton-bound organic matter to understand how nitrogen cycling in the coastal GBR has responded to increased anthropogenic nutrient inputs. Our major robust finding is that the coral record shows a long-term decline in skeletal 15N/14N towards the present. We argue that this decline is evidence for increased coastal nitrogen fixation rather than a direct reflection of anthropogenic nitrogen inputs. Reducing phosphorus discharge and availability would short-circuit the nitrogen fixation feedback loop and help avoid future acute and chronic eutrophication in the coastal GBR.

Pinto peanut cover crop nitrogen contributions and potential to mitigate nitrous oxide emissions in subtropical coffee plantations.

Many coffee (Coffea arabica L) production systems are characterised by high use of nitrogen (N) fertilisers, which can result in N leaching and emissions of nitrous oxide (N2O). We investigated the potential for legume cover crops grown inter-row to provide N for coffee trees and lower seasonal N2O emissions compared to poultry litter amendment at two subtropical field sites over 12 months, with unfertilised traditional grass groundcover used as a control treatment. Groundcovers (legume and grass treatments) were slashed from the inter-row into the tree line every 2-6 weeks as per normal farming operations. The prostrate ground cover legume Pinto peanut (Arachis pintoi) produced 4-5 t ha-1 biomass at both sites over the 12 month period, and fixed 146 kg N ha-1 year-1 at one site as estimated using the 15N natural abundance method. Background emissions from soil were lower at site 1 (0.38 kg N2O-N ha-1 year-1) than site 2 (2.26 kg N2O-N ha-1 year-1) reflecting differences in soil N and C levels at the sites. The use of Pinto peanut residues as an N amendment didn't result in any N2O flux events beyond those observed in the traditional groundcover control treatment across the season at either site, while the application of poultry litter to match farmer practice at these sites led to a major emission event. Ultimately, the Pinto peanut cover crop treatment led to a lower emission factor than for poultry litter at both sites, and resulted in significantly lower cumulative seasonal emissions for the legume cover crop (0.34 kg N2O-N ha-1 season-1) than poultry litter amendment (0.68 kg N2O-N ha-1 season-1) at site 1 despite similar inputs of N into the system. These findings suggest cover crop legumes could be integrated into coffee plantations to offset a portion of external N inputs, while lowering N2O emissions.

Effects of free nitrous acid treatment conditions on the nitrite pathway performance in mainstream wastewater treatment.

Inline sludge treatment using free nitrous acid (FNA) was recently shown to be effective in establishing the nitrite pathway in a biological nitrogen removal system. However, the effects of FNA treatment conditions on the nitrite pathway performance remained to be investigated. In this study, three different FNA treatment frequencies (daily sludge treatment ratios of 0.22, 0.31 and 0.38, respectively), two FNA concentrations (1.35 mgN/L and 4.23 mgN/L, respectively) and two influent feeding regimes (one- and two-step feeding) were investigated in four laboratory-scale sequencing batch reactors. The nitrite accumulation ratio was positively correlated to the FNA treatment frequency. However, when a high treatment frequency was used e.g., daily sludge treatment ratio of 0.38, a significant reduction in ammonia oxidizing bacteria (AOB) activity occurred, leading to poor ammonium oxidation. AOB were able to acclimatise to FNA concentrations up to of 4.23 mgN/L, whereas nitrite oxidizing bacteria (NOB) were limited by an FNA concentration of 1.35 mgN/L over the duration of the study (up to 120 days). This difference in sensitivity to FNA could be used to further enhance nitrite accumulation, with 90% accumulation achieved at an FNA concentration of 4.23 mgN/L and a daily sludge treatment ratio of 0.31 in this study. However, this high level of nitrite accumulation led to increased N2O emission, with emission factors of up to 3.9% observed. The N2O emission was mitigated (reduced to 1.3%) by applying two-step feeding resulting in a nitrite accumulation ratio of 45.1%. Economic analysis showed that choosing the optimal FNA treatment conditions depends on a combination of the wastewater characteristics, the nitrogen discharge standards, and the operational costs. This study provides important information for the optimisation and practical application of FNA-based sludge treatment technology for achieving the mainstream stable nitrite pathway.

Methane emissions partially offset "blue carbon" burial in mangroves.

Organic matter burial in mangrove forests results in the removal and long-term storage of atmospheric CO2, so-called "blue carbon." However, some of this organic matter is metabolized and returned to the atmosphere as CH4. Because CH4 has a higher global warming potential than the CO2 fixed in the organic matter, it can offset the CO2 removed via carbon burial. We provide the first estimate of the global magnitude of this offset. Our results show that high CH4 evasion rates have the potential to partially offset blue carbon burial rates in mangrove sediments on average by 20% (sensitivity analysis offset range, 18 to 22%) using the 20-year global warming potential. Hence, mangrove sediment and water CH4 emissions should be accounted for in future blue carbon assessments.

Quantifying nitrous oxide production pathways in wastewater treatment systems using isotope technology - A critical review.

Nitrous oxide (N2O) is an important greenhouse gas and an ozone-depleting substance which can be emitted from wastewater treatment systems (WWTS) causing significant environmental impacts. Understanding the N2O production pathways and their contribution to total emissions is the key to effective mitigation. Isotope technology is a promising method that has been applied to WWTS for quantifying the N2O production pathways. Within the scope of WWTS, this article reviews the current status of different isotope approaches, including both natural abundance and labelled isotope approaches, to N2O production pathways quantification. It identifies the limitations and potential problems with these approaches, as well as improvement opportunities. We conclude that, while the capabilities of isotope technology have been largely recognized, the quantification of N2O production pathways with isotope technology in WWTS require further improvement, particularly in relation to its accuracy and reliability.

Nitrous oxide fluxes in estuarine environments: response to global change.

Nitrous oxide is a powerful, long-lived greenhouse gas, but we know little about the role of estuarine areas in the global N2 O budget. This review summarizes 56 studies of N2 O fluxes and associated biogeochemical controlling factors in estuarine open waters, salt marshes, mangroves, and intertidal sediments. The majority of in situ N2 O production occurs as a result of sediment denitrification, although the water column contributes N2 O through nitrification in suspended particles. The most important factors controlling N2 O fluxes seem to be dissolved inorganic nitrogen (DIN) and oxygen availability, which in turn are affected by tidal cycles, groundwater inputs, and macrophyte density. The heterogeneity of coastal environments leads to a high variability in observations, but on average estuarine open water, intertidal and vegetated environments are sites of a small positive N2 O flux to the atmosphere (range 0.15-0.91; median 0.31; Tg N2 O-N yr(-1) ). Global changes in macrophyte distribution and anthropogenic nitrogen loading are expected to increase N2 O emissions from estuaries. We estimate that a doubling of current median NO3 (-) concentrations would increase the global estuary water-air N2 O flux by about 0.45 Tg N2 O-N yr(-1) or about 190%. A loss of 50% of mangrove habitat, being converted to unvegetated intertidal area, would result in a net decrease in N2 O emissions of 0.002 Tg N2 O-N yr(-1) . In contrast, conversion of 50% of salt marsh to unvegetated area would result in a net increase of 0.001 Tg N2 O-N yr(-1) . Decreased oxygen concentrations may inhibit production of N2 O by nitrification; however, sediment denitrification and the associated ratio of N2 O:N2 is expected to increase.

Behaviour of estrogenic endocrine-disrupting chemicals in permeable carbonate sands.

The remediation of four estrogenic endocrine-disrupting compounds (EDCs), estrone (E1), estradiol (E2), ethinylestradiol (EE2) and estriol (E3), was measured in saturated and unsaturated carbonate sand-filled columns dosed with wastewater from a sewage treatment plant. The estrogen equivalency (EEQ) of inlet wastewater was 1.2 ng L(-1) and was remediated to an EEQ of 0.5 ng L(-1) through the unsaturated carbonate sand-filled columns. The high surface area of carbonate sand and associated high microbial activity may have assisted the degradation of these estrogens. The fully saturated sand columns showed an increase in total estrogenic potency with an EEQ of 2.4 ng L(-1), which was double that of the inlet wastewater. There was a significant difference (P < 0.05) in total estrogenic potency between aerobic and anaerobic columns. The breakdown of conjugated estrogens to estrogenic EDCs formed under long residence time and reducing conditions may have been responsible for the increase in the fully saturated columns. This may also be explained by the desorption of previously sorbed estrogenic EDCs. The effect of additional filter materials, such as basalt sediment and coconut fibre, on estrogenic EDC reduction was also tested. None of these amendments provided improvements in estrogen remediation relative to the unamended unsaturated carbonate sand columns. Aerobic carbonate sand filters have good potential to be used as on-site wastewater treatment systems for the reduction of estrogenic EDCs. However, the use of fully saturated sand filters, which are used to promote denitrification, and the loss of nitrogen as N2 were shown to cause an increase in EEQ. The potential for the accumulation of estrogenic EDCs under anaerobic conditions needs to be considered when designing on-site sand filtration systems required to reduce nitrogen. Furthermore, the accumulation of estrogens under anaerobic conditions such as under soil absorption systems or leachate fields has the potential to contaminate groundwater especially when the water table levels fluctuate.

The effect of dissolved oxygen on N2O production by ammonia-oxidizing bacteria in an enriched nitrifying sludge.

Dissolved oxygen (DO) is commonly recognized as an important factor influencing nitrous oxide (N2O) production by ammonia-oxidizing bacteria (AOB). However, it has been difficult to separate the true effect of DO from that of nitrite, as DO variation often affects nitrite accumulation. The effect of DO on N2O production by an enriched nitrifying sludge, consisting of both AOB and nitrite-oxidizing bacteria (NOB), was investigated in this study. Nitrite accumulation was minimised by augmenting nitrite oxidation through the addition of an enriched NOB sludge. It was demonstrated that the specific N2O production rate increased from 0 to 1.9 ± 0.09 (n = 3) mg N2O-N/hr/g VSS with an increase of DO concentration from 0 to 3.0 mg O2/L, whereas N2O emission factor (the ratio between N2O nitrogen emitted and the ammonium nitrogen converted) decreased from 10.6 ± 1.7% (n = 3) at DO = 0.2 mg O2/L to 2.4 ± 0.1% (n = 3) at DO = 3.0 mg O2/L. The site preference measurements indicated that both the AOB denitrification and hydroxylamine (NH2OH) oxidation pathways contributed to N2O production, and DO had an important effect on the relative contributions of the two pathways. This finding is supported by analysis of the process data using an N2O model describing both pathways. As DO increased from 0.2 to 3.0 mg O2/L, the contribution of AOB denitrification decreased from 92% - 95%-66% - 73%, accompanied by a corresponding increase in the contribution by the NH2OH oxidation pathway.

Novel use of cavity ring-down spectroscopy to investigate aquatic carbon cycling from microbial to ecosystem scales.

Development of cavity ring-down spectroscopy (CRDS) has enabled real-time monitoring of carbon stable isotope ratios of carbon dioxide and methane in air. Here we demonstrate that CRDS can be adapted to assess aquatic carbon cycling processes from microbial to ecosystem scales. We first measured in situ isotopologue concentrations of dissolved CO2 ((12)CO2 and (13)CO2) and CH4 ((12)CH4 and (13)CH4) with CRDS via a closed loop gas equilibration device during a survey along an estuary and during a 40 h time series in a mangrove creek (ecosystem scale). A similar system was also connected to an in situ benthic chamber in a seagrass bed (community scale). Finally, a pulse-chase isotope enrichment experiment was conducted by measuring real-time release of (13)CO2 after addition of (13)C enriched phytoplankton to exposed intertidal sediments (microbial scale). Miller-Tans plots revealed complex transformation pathways and distinct isotopic source values of CO2 and CH4. Calculations of δ(13)C-DIC based on CRDS measured δ(13)C-CO2 and published fractionation factors were in excellent agreement with measured δ(13)C-DIC using isotope ratio mass spectroscopy (IRMS). The portable CRDS instrumentation used here can obtain real-time, high precision, continuous greenhouse gas data in lakes, rivers, estuaries and marine waters with less effort than conventional laboratory-based techniques.

Denitrification and anammox in tropical aquaculture settlement ponds: an isotope tracer approach for evaluating N2 production.

Settlement ponds are used to treat aquaculture discharge water by removing nutrients through physical (settling) and biological (microbial transformation) processes. Nutrient removal through settling has been quantified, however, the occurrence of, and potential for microbial nitrogen (N) removal is largely unknown in these systems. Therefore, isotope tracer techniques were used to measure potential rates of denitrification and anaerobic ammonium oxidation (anammox) in the sediment of settlement ponds in tropical aquaculture systems. Dinitrogen gas (N(2)) was produced in all ponds, although potential rates were low (0-7.07 nmol N cm(-3) h(-1)) relative to other aquatic systems. Denitrification was the main driver of N(2) production, with anammox only detected in two of the four ponds. No correlations were detected between the measured sediment variables (total organic carbon, total nitrogen, iron, manganese, sulphur and phosphorous) and denitrification or anammox. Furthermore, denitrification was not carbon limited as the addition of particulate organic matter (paired t-Test; P = 0.350, n = 3) or methanol (paired t-Test; P = 0.744, n = 3) did not stimulate production of N(2). A simple mass balance model showed that only 2.5% of added fixed N was removed in the studied settlement ponds through the denitrification and anammox processes. It is recommended that settlement ponds be used in conjunction with additional technologies (i.e. constructed wetlands or biological reactors) to enhance N(2) production and N removal from aquaculture wastewater.

Observations of nitrogen and phosphorus biogeochemistry in a surface flow constructed wetland.

Free surface water constructed wetlands (CWs) provide a buffer between domestic wastewater treatment plants and natural waterways. Understanding the biogeochemical processes in CWs is crucial to improve their performance. In this study we measured a range of water and sediment parameters, and biogeochemical processes, in an effort to describe the processing of nutrients within two wetland cells in series. As a whole the studied CW effectively absorbed both nitrogen (N) and phosphorus (P) emanating from the waste treatment plant. However the two individual cells showed marked differences related to the availability of oxygen within the water column and the sediments. In one cell we speculated that the prevalence of surface plant species reduced its ability to function as a net nutrient sink. Here we observed a build-up of sediment organic matter, sediment anoxia, a decoupling of nitrification-denitrification, and a flux of N and P out of the sediments to the overlying water. The availability of DO in the surface sediments of the second studied cell led to improved coupling between nitrification-denitrification and a net uptake of both NH4+ and PO4(3-). We hypothesise that the dominance of deeply rooted macrophytes in the second cell was responsible for the improved sediment quality.