High methane concentrations in tidal salt marsh soils: Where does the methane go?

Tidal salt marshes produce and emit CH4 . Therefore, it is critical to understand the biogeochemical controls that regulate CH4 spatial and temporal dynamics in wetlands. The prevailing paradigm assumes that acetoclastic methanogenesis is the dominant pathway for CH4 production, and higher salinity concentrations inhibit CH4 production in salt marshes. Recent evidence shows that CH4 is produced within salt marshes via methylotrophic methanogenesis, a process not inhibited by sulfate reduction. To further explore this conundrum, we performed measurements of soil-atmosphere CH4 and CO2 fluxes coupled with depth profiles of soil CH4 and CO2 pore water gas concentrations, stable and radioisotopes, pore water chemistry, and microbial community composition to assess CH4 production and fate within a temperate tidal salt marsh. We found unexpectedly high CH4 concentrations up to 145,000 µmol mol-1 positively correlated with S2- (salinity range: 6.6-14.5 ppt). Despite large CH4 production within the soil, soil-atmosphere CH4 fluxes were low but with higher emissions and extreme variability during plant senescence (84.3 ± 684.4 nmol m-2 s-1 ). CH4 and CO2 within the soil pore water were produced from young carbon, with most Δ14 C-CH4 and Δ14 C-CO2 values at or above modern. We found evidence that CH4 within soils was produced by methylotrophic and hydrogenotrophic methanogenesis. Several pathways exist after CH4 is produced, including diffusion into the atmosphere, CH4 oxidation, and lateral export to adjacent tidal creeks; the latter being the most likely dominant flux. Our findings demonstrate that CH4 production and fluxes are biogeochemically heterogeneous, with multiple processes and pathways that can co-occur and vary in importance over the year. This study highlights the potential for high CH4 production, the need to understand the underlying biogeochemical controls, and the challenges of evaluating CH4 budgets and blue carbon in salt marshes.

Las marismas salinas producen y emiten CH4 . Por lo tanto, es esencial comprender los controles biogeoquímicos que regulan la dinámica espacial y temporal del CH4 en estos humedales. El paradigma predominante asume que la metanogénesis acetoclástica es la vía dominante para la producción de CH4 y que altas concentraciones de salinidad inhiben la producción de CH4 en estos ecosistemas. Hay evidencia que el CH4 se produce las marismas salinas a través de la metanogénesis metilotrófica, un proceso no inhibido por la reducción del sulfato. Para explorar esta paradoja, realizamos mediciones de los flujos de CH4 y CO2 del suelo a la atmósfera junto con perfiles de concentraciones de CH4 y CO2 en el suelo, isótopos estables y radioisótopos, química del agua y composición de la comunidad microbiana para evaluar la producción y el destino del CH4 en una marisma salina templada. Encontramos concentraciones de CH4 sorprendentemente altas de hasta 145,000 µmol mol−1 correlacionadas positivamente con S2− (rango de salinidad: 6.6 a 14.5 ppt). A pesar de la gran producción de CH4 en el suelo, los flujos de CH4 del suelo a la atmósfera fueron bajos, pero con mayores emisiones y variabilidad extrema durante la época de senescencia de las plantas (84.3 ± 684.4 nmol m−2 s−1 ). El CH4 y el CO2 en el suelo se produjeron a partir de carbono joven, con la mayoría de los valores Δ14 C-CH4 y Δ14 C-CO2 en o por encima de valores modernos. Encontramos evidencia de que el CH4 en los suelos fue producido por metanogénesis metilotrófica e hidrogenotrófica. Existen varias vías que el CH4 producido sigue, incluida la difusión hacia la atmósfera, la oxidación del CH4 y la exportación lateral a arroyos adyacentes a la marisma; siendo este último el flujo dominante más probable. Nuestros hallazgos demuestran que la producción y los flujos de CH4 son biogeoquímicamente heterogéneos, con múltiples procesos y vías que pueden coexistir y variar en importancia a lo largo del año. Este estudio destaca el potencial de alta producción de CH4 , la necesidad de comprender los controles biogeoquímicos de la producción de CH4 y los retos que existen para evaluar las reservas de CH4 y el carbono azul en marismas salinas.

Adsorption and Removal Kinetics of 2,4-Dinitroanisole and Nitrotriazolone in Contrasting Freshwater Sediments: Batch Study.

Environmental release of 2,4-dinitroanisole (DNAN) and 3-nitro-1,2,4-triazol-5-one (NTO) is of great concern due to high migration potential in the environment. In the present study we evaluated the adsorption and microbially-mediated removal kinetics of dissolved DNAN and NTO in contrasting freshwater sediments with different total organic carbon (TOC) content. River sand (low TOC), pond silt (high TOC), clay-rich lake sediment (low TOC), wetland silt (high TOC), carbonate sand (low TOC), and iron-rich clay (low TOC) were evaluated. Separate abiotic and biotic bench-top sediment slurry incubations were carried out at 23, 15, and 4 °C for DNAN and NTO. Experiments were conducted over 3 weeks. Time series aqueous samples and sediment samples collected at the end of the experiment were analyzed for DNAN and NTO concentrations. The DNAN compound equilibrated with sediment within the first 2 h after addition whereas NTO showed no adsorption. 2,4-Dinitroanisole adsorbed more onto fine-grained organic-rich sediments (Kd = 2-40 L kg-1 sed-1 ) than coarse-grained organic-poor sediments (Kd = 0.2-0.6 L kg-1 sed-1 ), and the TOC content and cation exchange capacity of sediment were reliable predictors for abiotic DNAN adsorption. Adsorption rate constants and equilibrium partitioning constants for DNAN were inversely proportional to temperature in all sediment types. The biotic removal half-life of DNAN was faster (t1/2 = 0.1-58 h) than that of NTO (t1/2 = 5-347 h) in all sediment slurries. Biotic removal rates (t1/2 = 0.1-58 h) were higher than abiotic rates (t1/2 = 0.3-107 h) for DNAN at 23 °C. Smaller grain size coupled with higher TOC content enhanced biotic NTO and DNAN removal in freshwater environments. Environ Toxicol Chem 2023;42:46-59. © 2022 SETAC.

A grazing-driven positive nutrient feedback loop and active sexual reproduction underpin widespread Noctiluca green tides.

The mixoplankton green Noctiluca scintillans (gNoctiluca) is known to form extensive green tides in tropical coastal ecosystems prone to eutrophication. In the Arabian Sea, their recent appearance and annual recurrence have upended an ecosystem that was once exclusively dominated by diatoms. Despite evidence of strong links to eutrophication, hypoxia and warming, the mechanisms underlying outbreaks of this mixoplanktonic dinoflagellate remain uncertain. Here we have used eco-physiological measurements and transcriptomic profiling to ascribe gNoctiluca's explosive growth during bloom formation to the form of sexual reproduction that produces numerous gametes. Rapid growth of gNoctiluca coincided with active ammonium and phosphate release from gNoctiluca cells, which exhibited high transcriptional activity of phagocytosis and metabolism generating ammonium. This grazing-driven nutrient flow ostensibly promotes the growth of phytoplankton as prey and offers positive support successively for bloom formation and maintenance. We also provide the first evidence that the host gNoctiluca cell could be manipulating growth of its endosymbiont population in order to exploit their photosynthetic products and meet critical energy needs. These findings illuminate gNoctiluca's little known nutritional and reproductive strategies that facilitate its ability to form intense and expansive gNoctiluca blooms to the detriment of regional water, food and the socio-economic security in several tropical countries.

Ecosystem-based management for military training, biodiversity, carbon storage and climate resiliency on a complex coastal land/water-scape.

The Defense Coastal/Estuarine Research Program (DCERP) was a 10-year multi-investigator project funded by the Department of Defense to improve understanding of ecosystem processes and their interactions with natural and anthropogenic stressors at the Marine Corps Base Camp Lejeune (MCBCL) located in coastal North Carolina. The project was aimed at facilitating ecosystem-based management (EBM) at the MCBCL and other coastal military installations. Because of its scope, interdisciplinary character, and duration, DCERP embodied many of the opportunities and challenges associated with EBM, including the need for explicit goals, system models, long-term perspectives, systems complexity, change inevitability, consideration of humans as ecosystem components, and program adaptability and accountability. We describe key elements of this program, its contributions to coastal EBM, and its relevance as an exemplar of EBM.

Degradation of RDX (Hexahydro-1,3,5-trinitro-1,3,5-triazine) in contrasting coastal marine habitats: Subtidal non-vegetated (sand), subtidal vegetated (silt/eel grass), and intertidal marsh.

Hundreds of explosive-contaminated marine sites exist globally, many of which contain the common munitions constituent hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). Quantitative information about RDX transformation in coastal ecosystems is essential for management of many of these sites. Isotopically labelled RDX containing 15N in all 3 nitro groups was used to track the fate of RDX in three coastal ecosystem types. Flow-through mesocosms representing subtidal vegetated (silt/eel grass), subtidal non-vegetated (sand) and intertidal marsh ecosystems were continuously loaded with isotopically labelled RDX for 16-17 days. Sediment, pore-water and overlying surface water were analyzed to determine the distribution of RDX, nitroso-triazine transformation products (NXs) and nitrogen containing complete mineralization products, including ammonium, nitrate+nitrite, nitrous oxide and nitrogen gas. The marsh, silt, and sand ecotypes transformed 94%, 90% and 76% of supplied RDX, respectively. Total dissolved NXs accounted for 2%-4% of the transformed 15N-RDX. The majority of RDX transformation in the water column was by mineralization to inorganic N (dissolved and evaded; 64%-78% of transformed 15N-RDX). RDX was mineralized primarily to N2O (62-74% of transformed 15N-RDX) and secondarily to N2 (1-2% of transformed 15N-RDX) which exchanged with the atmosphere. Transformation of RDX was favored in carbon-rich lower redox potential sediments of the silt and marsh mesocosms where anaerobic processes of iron and sulfate reduction were most prevalent. RDX was most persistent in the carbon-poor sand mesocosm. Partitioning of 15N derived from RDX onto sediment and suspended particulates was negligible in the overall mass balance of RDX transformation (2%-3% of transformed 15N-RDX). The fraction of 15N derived from RDX that was sorbed or assimilated in sediment was largest in the marsh mesocosm (most organic carbon), and smallest in the sand mesocosm (largest grain size and least organic carbon). Sediment redox conditions and available organic carbon stores affect the fate of RDX in different coastal marine habitats.

The influence of sample matrix on the accuracy of nitrite N and O isotope ratio analyses with the azide method.

RATIONALE: The isotope ratios of nitrogen (15 N/14 N) and oxygen (18 O/16 O) in nitrite (NO2 - ) can be measured by conversion of the nitrite into nitrous oxide (N2 O) with azide, followed by mass spectrometric analysis of N2 O by gas chromatography isotope ratio mass spectrometry (GC/IRMS). While applying this method to brackish samples, we noticed that the N and O isotope ratio measurements of NO2 - are highly sensitive to sample salinity and to the pH at which samples are preserved. METHODS: We investigated the influence of sample salinity and sample preservation pH on the N and O isotope ratios of the N2 O produced from the reaction of NO2 - with azide. The N2 O isotope ratios were measured by GC/IRMS. RESULTS: Under the experimental reaction conditions, the conversion of NO2 - into N2 O was less complete in lower salinity solutions, resulting in respective N and O isotopic offsets of +2.5‰ and -14.0‰ compared with seawater solutions. Differences in salinity were also associated with differences in the fraction of O atoms exchanged between NO2 - and water during the reaction. Similarly, aqueous NO2 - samples preserved at elevated pH values resulted in the incomplete conversion of NO2 - into N2 O by azide, and consequent pH-dependent isotopic offsets, as well as differences in the fraction of O atoms exchanged with water. The addition of sodium chloride to the reaction matrix of samples and standards largely mitigated salinity-dependent isotopic offsets in the N2 O product, and nearly homogenized the fraction of O atom exchange among samples of different salinity. A test of the hypobromite-azide method to measure N isotope ratios of ammonium by conversion into NO2 - then N2 O revealed no influence of sample salinity on the N isotope ratios of the N2 O product. CONCLUSIONS: We outline recommendations to mitigate potential matrix effects among samples and standards, to improve the accuracy of N and O isotope ratios in NO2 - measured with the azide method.

Constraining the Oxygen Isotopic Composition of Nitrate Produced by Nitrification.

Measurements of the stable isotope ratios of nitrogen (15N/14N) and oxygen (18O/16O) in nitrate (NO3-) enable identification of sources, dispersal, and fate of natural and contaminant NO3- in aquatic environments. The 18O/16O of NO3- produced by nitrification is often assumed to reflect the proportional contribution of oxygen atom sources, water, and molecular oxygen, in a 2:1 ratio. Culture and seawater incubations, however, indicate oxygen isotopic equilibration between nitrite (NO2-) and water, and kinetic isotope effects for oxygen atom incorporation, which modulate the NO3- 18O/16O produced during nitrification. To investigate the influence of kinetic and equilibrium effects on the isotopic composition of NO3- produced from the nitrification of ammonia (NH3), we incubated streamwater supplemented with ammonium (NH4+) and increments of 18O-enriched water. Resulting NO3- 18O/16O ratios showed (1) a disproportionate sensitivity to the 18O/16O ratio of water, mediated by isotopic equilibration between water and NO2-, as well as (2) kinetic isotope discrimination during O atom incorporation from molecular oxygen and water. Empirically, the NO3- 18O/16O ratios thus produced fortuitously converge near the 18O/16O ratio of water. More elevated NO3- 18O/16O values commonly reported in soils and oxic groundwater may thus derive from processes additional to nitrification, including NO3- reduction.

Tracing the cycling and fate of the munition, Hexahydro-1,3,5-trinitro-1,3,5-triazine in a simulated sandy coastal marine habitat with a stable isotopic tracer, 15N-[RDX].

Coastal marine habitats become contaminated with the munitions constituent, Hexahydro-1,3,5-trinitro-1,3,5-trazine (RDX), via military training, weapon testing and leakage of unexploded ordnance. This study used 15N labeled RDX in simulated aquarium-scale coastal marine habitat containing seawater, sediment, and biota to track removal pathways from surface water including sorption onto particulates, degradation to nitroso-triazines and mineralization to dissolved inorganic nitrogen (DIN). The two aquaria received continuous RDX inputs to maintain a steady state concentration (0.4 mg L-1) over 21 days. Time series RDX and nitroso-triazine concentrations in dissolved (surface and porewater) and sorbed phases (sediment and suspended particulates) were analyzed. Distributions of DIN species (ammonium, nitrate + nitrite and dissolved N2) in sediments and overlying water were also measured along with geochemical variables in the aquaria. Partitioning of RDX and RDX-derived breakdown products onto surface sediment represented 13% of the total added 15N as RDX (15N-[RDX]) equivalents after 21 days. Measured nitroso-triazines in the aquaria accounted for 6-13% of total added 15N-[RDX]. 15N-labeled DIN was found both in the oxic surface water and hypoxic porewaters, showing that RDX mineralization accounted for 34% of the 15N-[RDX] added to the aquaria over 21 days. Labeled ammonium (15NH4+, found in sediment and overlying water) and nitrate + nitrite (15NOX, found in overlying water only) together represented 10% of the total added 15N-[RDX]. The production of 15N labeled N2 (15N2), accounted for the largest individual sink during the transformation of the total added 15N-[RDX] (25%). Hypoxic sediment was the most favorable zone for production of N2, most of which diffused through porous sediments into the water column and escaped to the atmosphere.

Investigation of a new passive sampler for the detection of munitions compounds in marine and freshwater systems.

Over the last century, unexploded ordnances have been disposed of in marine shelf systems because of a lack of cost-effective alternatives. Underwater unexploded ordnances have the potential to leak 2,4,6-trinitrotoluene (TNT) and 1,3,5-trinitro-1,3,5-triazine (RDX), commonly used chemical munitions, and contaminate local waters, biota, and sediments. The rate at which this contamination occurs in the environment is relatively unknown, and the cost- and time-prohibitive nature of sampling across sites makes mapping difficult. In the present study we assessed the efficacy of ethylene-vinyl acetate (EVA) for sampling relatively soluble munitions compounds over a range of environmental conditions (i.e., changes in temperature and salinity) and optimized the composition of the passive sampling polymer. The EVA sampler was able to successfully detect ambient concentrations of lingering munitions compounds from field sites containing unexploded ordnances. The sampler affinity for the munitions in terms of an EVA-water partition coefficient was greater than the standard octanol water values for each target compound. Partitioning of compounds onto EVA over the natural ranges of salinity did not change significantly, although uptake varied consistently and predictably with temperature. Increasing the vinyl acetate to ethylene ratio of the polymer corresponded to an increase in uptake capacity, consistent with enhanced dipole-dipole interactions between the munitions and the polymer. This sampler provides a cost-effective means to map and track leakage of unexploded ordnances both spatially and temporally. Environ Toxicol Chem 2018;37:1990-1997. © 2018 SETAC.

Corrigendum: Chemical formation of hybrid di-nitrogen calls fungal codenitrification into question.

This corrects the article DOI: 10.1038/srep39077.

Biodegradation and mineralization of isotopically labeled TNT and RDX in anaerobic marine sediments.

The lack of knowledge on the fate of explosive compounds 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), particularly in marine ecosystems, constrains the application of bioremediation techniques in explosive-contaminated coastal sites. The authors present a comparative study on anaerobic biodegradation and mineralization of 15 N-nitro group isotopically labeled TNT and RDX in organic carbon-rich, fine-grained marine sediment with native microbial assemblages. Separate sediment slurry experiments were carried out for TNT and RDX at 23°C for 16 d. Dissolved and sediment-sorbed fractions of parent and transformation products, isotopic compositions of sediment, and mineralization products of the dissolved inorganic N pool (15 NH4+ ,15 NO3- ,15 NO2- , and 15 N2 ) were measured. The rate of TNT removal from the aqueous phase was faster (0.75 h-1 ) than that of RDX (0.37 h-1 ), and 15 N accumulation in sediment was higher in the TNT (13%) than the RDX (2%) microcosms. Mono-amino-dinitrotoluenes were identified as intermediate biodegradation products of TNT. Two percent of the total spiked TNT-N is mineralized to dissolved inorganic N through 2 different pathways: denitration as well as deamination and formation of NH4+ , facilitated by iron and sulfate reducing bacteria in the sediments. The majority of the spiked TNT-N (85%) is in unidentified pools by day 16. Hexahydro-1,3,5-trinitro-1,3,5-triazine (10%) biodegrades to nitroso derivatives, whereas 13% of RDX-N in nitro groups is mineralized to dissolved inorganic N anaerobically by the end of the experiment. The primary identified mineralization end product of RDX (40%) is NH4+ , generated through either deamination or mono-denitration, followed by ring breakdown. A reasonable production of N2 gas (13%) was seen in the RDX system but not in the TNT system. Sixty-eight percent of the total spiked RDX-N is in an unidentified pool by day 16 and may include unquantified mineralization products dissolved in water. Environ Toxicol Chem 2017;36:1170-1180. © 2016 SETAC.

Chemical formation of hybrid di-nitrogen calls fungal codenitrification into question.

Removal of excess nitrogen (N) can best be achieved through denitrification processes that transform N in water and terrestrial ecosystems to di-nitrogen (N2) gas. The greenhouse gas nitrous oxide (N2O) is considered an intermediate or end-product in denitrification pathways. Both abiotic and biotic denitrification processes use a single N source to form N2O. However, N2 can be formed from two distinct N sources (known as hybrid N2) through biologically mediated processes of anammox and codenitrification. We questioned if hybrid N2 produced during fungal incubation at neutral pH could be attributed to abiotic nitrosation and if N2O was consumed during N2 formation. Experiments with gas chromatography indicated N2 was formed in the presence of live and dead fungi and in the absence of fungi, while N2O steadily increased. We used isotope pairing techniques and confirmed abiotic production of hybrid N2 under both anoxic and 20% O2 atmosphere conditions. Our findings question the assumptions that (1) N2O is an intermediate required for N2 formation, (2) production of N2 and N2O requires anaerobiosis, and (3) hybrid N2 is evidence of codenitrification and/or anammox. The N cycle framework should include abiotic production of N2.

Uptake and fate of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in coastal marine biota determined using a stable isotopic tracer, (15)N - [RDX].

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is globally one of the most commonly used military explosives and environmental contaminant. (15)N labeled RDX was added into a mesocosm containing 9 different coastal marine species in a time series experiment to quantify the uptake of RDX and assess the RDX derived (15)N retention into biota tissue. The (15)N attributed to munitions compounds reached steady state concentrations ranging from 0.04 to 0.67 µg (15)N g dw(-1), the bulk (15)N tissue concentration for all species was 1-2 orders of magnitude higher suggesting a common mechanism or pathway of RDX biotransformation and retention of (15)N. A toxicokinetic model was created that described the (15)N uptake, elimination, and transformation rates. While modeled uptake rates were within previous published values, elimination rates were several orders of magnitude smaller than previous studies ranging from 0.05 to 0.7 days(-1). These small elimination rates were offset by high rates of retention of (15)N previously not measured. Bioconcentration factors and related aqueous:organism ratios of compounds and tracer calculated using different tracer and non-tracer methods yielded a broad range of values (0.35-101.6 mL g(-1)) that were largely method dependent. Despite the method-derived variability, all values were generally low and consistent with little bioaccumulation potential. The use of (15)N labeled RDX in this study indicates four possible explanations for the observed distribution of compounds and tracer; each with unique potential implications for possible toxicological impacts in the coastal marine environment.

Impact of chloride on denitrification potential in roadside wetlands.

Developed landscapes are exposed to changes in hydrology and water chemistry that limit their ability to mitigate detrimental impacts to coastal water bodies, particularly those that result from stormwater runoff. The elevated level of impervious cover increases not only runoff but also contaminant loading of nutrients, metals, and road salt used for deicing to water bodies. Here we investigate the impact that road salt has on denitrification in roadside environments. Sediments were collected from a series of forested and roadside wetlands and acclimated with a range of Cl(-) concentrations from 0 to 5000 mg L(-1) for 96 h. Denitrification rates were measured by the isotope pairing technique using (15)N-NO3(-), while denitrifying community structures were compared using terminal restriction fragment length polymorphism (T-RFLP) of nitrous oxide reductase genes (nosZ). Chloride significantly (p < 0.05) inhibited denitrification in forested wetlands at a Cl(-) dosage of 2500 or 5000 mg L(-1), but the decrease in denitrification rates was less and not significant for the roadside wetlands historically exposed to elevated concentrations of Cl(-). The difference could not be attributed to other significant changes in conditions, such as DOC concentrations, N species concentrations, or pH levels. Denitrifying communities, as measured by T-RFs of the nosZ gene, in the roadside wetlands with elevated concentration of Cl(-) were distinctly different and more diverse compared to forested wetlands, and also different in roadside wetlands after 96 h exposures to Cl(-). The shifts in denitrifying communities seem to minimize the decrease in denitrification rates in the wetlands previously exposed to Cl. As development results in more Cl(-) use and exposure to a broad range of natural or manmade wetland structures, an understanding of the seasonal effect of Cl on denitrification processes in these systems would aid in design or mitigation of the effects on N removal rates.

Fungal denitrification: Bipolaris sorokiniana exclusively denitrifies inorganic nitrogen in the presence and absence of oxygen.

Fungi may play an important role in the production of the greenhouse gas nitrous oxide (N2O). Bipolaris sorokiniana is a ubiquitous saprobe found in soils worldwide, yet denitrification by this fungal strain has not previously been reported. We aimed to test if B. sorokiniana would produce N2O and CO2 in the presence of organic and inorganic forms of nitrogen (N) under microaerobic and anaerobic conditions. Nitrogen source (organic-N, inorganic-N, no-N control) significantly affected N2O and CO2 production both in the presence and absence of oxygen, which contrasts with bacterial denitrification. Inorganic N addition increased denitrification of N2O (from 0 to 0.3 µg N20-N h(-1) g(-1) biomass) and reduced respiration of CO2 (from 0.1 to 0.02 mg CO2 h(-1) g(-1) biomass). Isotope analyses indicated that nitrite, rather than ammonium or glutamine, was transformed to N2O. Results suggest the source of N may play a larger role in fungal N2O production than oxygen status.

Sorption kinetics of TNT and RDX in anaerobic freshwater and marine sediments: Batch studies.

Examination of the partitioning of explosives onto sediment in marine environments is critical to predict the toxicological impacts of worldwide explosive-contaminated sites adjacent to estuaries, wetlands, and the coastal ocean. Marine sediments have been identified as sites of enhanced munitions removal, yet most studies addressing these interactions focus on soils and freshwater sediments. The present study measured the kinetics of 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) sorption onto 2 marine sediments of varying grain sizes (silt vs sand) and organic carbon (OC) content. Abiotic sediment sorption tests were performed at 23 °C, 15 °C, and 4 °C by spiking TNT and RDX solutions directly into anaerobic sediment slurries. Marine sediments showed significantly higher compound uptake rates (0.30-0.80 h(-1) ) than freshwater silt (0.0046-0.0065 h(-1) ) for both compounds, probably because of lower compound solubilities and a higher pH in marine systems. Equilibrium partition constants are on the same order of magnitude for marine silt (1.1-2.0 L kg(-1) sediment) and freshwater silt (1.4-3.1 L kg(-1) sediment) but lower for marine sand (0.72-0.92 L kg(-1) sediment). Total organic carbon content in marine sediments varied linearly with equilibrium partition constants for TNT and was moderately linear for RDX. Uptake rates and equilibrium constants of explosives are inversely correlated to temperature regardless of sediment type because of kinetic barriers associated with low temperatures.

Role of Anaerobic Ammonium Oxidation (Anammox) in Nitrogen Removal from a Freshwater Aquifer.

Anaerobic ammonium oxidation (anammox) couples the oxidation of ammonium with the reduction of nitrite, producing N2. The presence and activity of anammox bacteria in groundwater were investigated at multiple locations in an aquifer variably affected by a large, wastewater-derived contaminant plume. Anammox bacteria were detected at all locations tested using 16S rRNA gene sequencing and quantification of hydrazine oxidoreductase (hzo) gene transcripts. Anammox and denitrification activities were quantified by in situ (15)NO2(-) tracer tests along anoxic flow paths in areas of varying ammonium, nitrate, and organic carbon abundances. Rates of denitrification and anammox were determined by quantifying changes in (28)N2, (29)N2, (30)N2, (15)NO3(-), (15)NO2(-), and (15)NH4(+) with groundwater travel time. Anammox was present and active in all areas tested, including where ammonium and dissolved organic carbon concentrations were low, but decreased in proportion to denitrification when acetate was added to increase available electron supply. Anammox contributed 39-90% of potential N2 production in this aquifer, with rates on the order of 10 nmol N2-N L(-1) day(-1). Although rates of both anammox and denitrification during the tracer tests were low, they were sufficient to reduce inorganic nitrogen concentrations substantially during the overall groundwater residence times in the aquifer. These results demonstrate that anammox activity in groundwater can rival that of denitrification and may need to be considered when assessing nitrogen mass transport and permanent loss of fixed nitrogen in aquifers.

Tracing the Cycling and Fate of the Explosive 2,4,6-Trinitrotoluene in Coastal Marine Systems with a Stable Isotopic Tracer, (15)N-[TNT].

2,4,6-Trinitrotoluene (TNT) has been used as a military explosive for over a hundred years. Contamination concerns have arisen as a result of manufacturing and use on a large scale; however, despite decades of work addressing TNT contamination in the environment, its fate in marine ecosystems is not fully resolved. Here we examine the cycling and fate of TNT in the coastal marine systems by spiking a marine mesocosm containing seawater, sediments, and macrobiota with isotopically labeled TNT ((15)N-[TNT]), simultaneously monitoring removal, transformation, mineralization, sorption, and biological uptake over a period of 16 days. TNT degradation was rapid, and we observed accumulation of reduced transformation products dissolved in the water column and in pore waters, sorbed to sediments and suspended particulate matter (SPM), and in the tissues of macrobiota. Bulk δ(15)N analysis of sediments, SPM, and tissues revealed large quantities of (15)N beyond that accounted for in identifiable derivatives. TNT-derived N was also found in the dissolved inorganic N (DIN) pool. Using multivariate statistical analysis and a (15)N mass balance approach, we identify the major transformation pathways of TNT, including the deamination of reduced TNT derivatives, potentially promoted by sorption to SPM and oxic surface sediments.

Mineralization of RDX-derived nitrogen to N2 via denitrification in coastal marine sediments.

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is a common constituent of military explosives. Despite RDX contamination at numerous U.S. military facilities and its mobility to aquatic systems, the fate of RDX in marine systems remains largely unknown. Here, we provide RDX mineralization pathways and rates in seawater and sediments, highlighting for the first time the importance of the denitrification pathway in determining the fate of RDX-derived N. (15)N nitro group labeled RDX ((15)N-[RDX], 50 atom %) was spiked into a mesocosm simulating shallow marine conditions of coastal Long Island Sound, and the (15)N enrichment of N2 (δ(15)N2) was monitored via gas bench isotope ratio mass spectrometry (GB-IRMS) for 21 days. The (15)N tracer data were used to model RDX mineralization within the context of the broader coastal marine N cycle using a multicompartment time-stepping model. Estimates of RDX mineralization rates based on the production and gas transfer of (15)N2O and (15)N2 ranged from 0.8 to 10.3 µmol d(-1). After 22 days, 11% of the added RDX had undergone mineralization, and 29% of the total removed RDX-N was identified as N2. These results demonstrate the important consideration of sediment microbial communities in management strategies addressing cleanup of contaminated coastal sites by military explosives.

Groundwater nitrogen processing in Northern Gulf of Mexico restored marshes.

Groundwater nitrogen processing was examined in a restored black needlerush (Juncus roemerianus) marsh to assess its potential for removing land-derived nitrogen pollution. Two restoration designs, one initially planted at 50% cover (half density plots) and the other one at 100% cover (full density plots), were compared with non-vegetated controls. The introduction via groundwater of a NO3(-) solution with a conservative tracer (Br(-)) and labeled isotopically ((15)N) allowed calculation of nitrogen removal in the plots following two methods. The first method used changes in the ratio [NOx]:[Br(-)] as the groundwater plume traveled through the plot, and the second method relied on balancing (15)N input with (15)N export. Both methods showed ≈97% of the N from the simulated groundwater plume was removed (i.e. not delivered to the open waters of the adjacent estuary) in vegetated plots and ≈86% was removed in non-vegetated controls. The most dominant routes of N removal from the introduced solution were N2 production and assimilation into macrophyte biomass, which were similar in magnitude for the vegetated plots, whereas N2 production dominated in the unvegetated plots. The majority of N removed from the introduced solution occurred in the first 30 cm the solution traveled in the vegetated treatments. In addition, ambient porewater concentrations of dissolved inorganic nitrogen (DIN) were similar between full and half density plots, but lower than the non-vegetated control (≈8.5× and 7.5×), suggesting full and half density plots removed more DIN than non-vegetated plots. These results suggest that restoring marshes by planting 50% of the area may be a more cost-effective restoration design in terms of mitigating land-derived nutrient pollution than planting 100% of the area since it requires less effort and cost while removing similar quantities of N.