Denise A. Bruesewitz

Nitrate removal, communities of denitrifiers and adverse effects in different carbon substrates for use in denitrification beds.

Denitrification beds are containers filled with wood by-products that serve as a carbon and energy source to denitrifiers, which reduce nitrate (NO(3)(-)) from point source discharges into non-reactive dinitrogen (N(2)) gas. This study investigates a range of alternative carbon sources and determines rates, mechanisms and factors controlling NO(3)(-) removal, denitrifying bacterial community, and the adverse effects of these substrates. Experimental barrels (0.2 m(3)) filled with either maize cobs, wheat straw, green waste, sawdust, pine woodchips or eucalyptus woodchips were incubated at 16.8 °C or 27.1 °C (outlet temperature), and received NO(3)(-) enriched water (14.38 mg N L(-1) and 17.15 mg N L(-1)). After 2.5 years of incubation measurements were made of NO(3)(-)-N removal rates, in vitro denitrification rates (DR), factors limiting denitrification (carbon and nitrate availability, dissolved oxygen, temperature, pH, and concentrations of NO(3)(-), nitrite and ammonia), copy number of nitrite reductase (nirS and nirK) and nitrous oxide reductase (nosZ) genes, and greenhouse gas production (dissolved nitrous oxide (N(2)O) and methane), and carbon (TOC) loss. Microbial denitrification was the main mechanism for NO(3)(-)-N removal. Nitrate-N removal rates ranged from 1.3 (pine woodchips) to 6.2 g N m(-3) d(-1) (maize cobs), and were predominantly limited by C availability and temperature (Q(10) = 1.2) when NO(3)(-)-N outlet concentrations remained above 1 mg L(-1). The NO(3)(-)-N removal rate did not depend directly on substrate type, but on the quantity of microbially available carbon, which differed between carbon sources. The abundance of denitrifying genes (nirS, nirK and nosZ) was similar in replicate barrels under cold incubation, but varied substantially under warm incubation, and between substrates. Warm incubation enhanced growth of nirS containing bacteria and bacteria that lacked the nosZ gene, potentially explaining the greater N(2)O emission in warmer environments. Maize cob substrate had the highest NO(3)(-)-N removal rate, but adverse effects include TOC release, dissolved N(2)O release and substantial carbon consumption by non-denitrifiers. Woodchips removed less than half of NO(3)(-) removed by maize cobs, but provided ideal conditions for denitrifying bacteria, and adverse effects were not observed. Therefore we recommend the combination of maize cobs and woodchips to enhance NO(3)(-) removal while minimizing adverse effects in denitrification beds.

A comparison of different approaches for measuring denitrification rates in a nitrate removing bioreactor.

Denitrifying woodchip bioreactors (denitrification beds) are increasingly used to remove excess nitrate (NO₃⁻) from point-sources such as wastewater effluent or subsurface drains from agricultural fields. NO₃⁻ removal in these beds is assumed to be due to microbial denitrification but direct measurements of denitrification are lacking. Our objective was to test four different approaches for measuring denitrification rates in a denitrification bed that treated effluent discharged from a glasshouse. We compared these denitrification rates with the rate of NO₃⁻ removal along the length of the bed. The NO₃⁻ removal rate was 8.73 ± 1.45 g m⁻³ d⁻¹. In vitro acetylene inhibition assays resulted in highly variable denitrification rates (DR(AI)) along the length of the bed and generally 5 times greater than the measured (NO₃⁻-N removal rate. An in situ push-pull test, where enriched ¹⁵N-NO₃⁻ was injected into 2 locations along the bed, resulted in rates of 23.2 ± 1.43 g N m⁻³ d⁻¹ and 8.06 ± 1.64 g N m⁻³ d⁻¹. The denitrification rate calculated from the increase in dissolved N₂ and N₂O concentrations (DR(N₂) along the length of the denitrification bed was 6.7 ± 1.61 g N m⁻³ d⁻¹. Lastly, denitrification rates calculated from changes in natural abundance measurements of δ¹⁵N-N₂ and δ¹⁵N-NO₃⁻ along the length of the bed yielded a denitrification rate (DR(NA)) of 6.39 ± 2.07 g m⁻³ d⁻¹. Based on our experience, DR(N₂) measurements were the easiest and most efficient approach for determining the denitrification rate and N₂O production of a denitrification bed. However, the other approaches were useful for testing other hypotheses such as factors limiting denitrification or may be applied to determine denitrification rates in environmental systems different to our study site. DR(N₂) does require very careful sampling to avoid atmospheric N₂ contamination but could be used to rapidly determine denitrification rates in a variety of aquatic systems with high N₂ production and even water flows. These measurements demonstrated that the majority of NO₃⁻ removal was due to heterotrophic denitrification.

Intra- and inter-annual variability in metabolism in an oligotrophic lake

Lakes are sentinels of change in the landscapes in which they are located. Changes in lake function are reflected in whole-system metabolism, which integrates ecosystem processes across spatial and temporal scales. Recent improvements in high-frequency open-water metabolism modeling techniques have enabled estimation of rates of gross primary production (GPP), respiration (R), and net ecosystem production (NEP) at high temporal resolution. However, few studies have examined metabolic rates over daily to multi-year temporal scales, especially in oligotrophic ecosystems. Here, we modified a metabolism modeling technique to reveal substantial intra- and inter-annual variability in metabolic rates in Lake Sunapee, a temperate, oligotrophic lake in New Hampshire, USA. Annual GPP and R increased each summer, paralleling increases in littoral, but not pelagic, total phosphorus concentrations. Storms temporarily decoupled GPP and R, resulting in greater decreases in GPP than R. Daily rates of GPP and R were positively correlated on warm days that had stable water columns, and metabolism model fits were best on warm, sunny days, indicating the importance of lake physics when evaluating metabolic rates. These metabolism data span a range of temporal scales and together suggest that Lake Sunapee may be moving toward mesotrophy. We suggest that functional, integrative metrics, such as metabolic rates, are useful indicators and sentinels of ecosystem change. We also highlight the challenges and opportunities of using high-frequency measurements to elucidate the drivers and consequences of intra- and inter-annual variability in metabolic rates, especially in oligotrophic lakes.

Seasonal Water Column NH4 + Cycling Along a Semi-arid Sub-tropical River–Estuary Continuum: Responses to Episodic Events and Drought Conditions

AbstractRiver–estuary continuums represent a dynamic range of environmental conditions in aquatic ecosystems, providing an ideal gradient for understanding changes in nitrogen (N) cycling. We measured rates of ammonium (NH4+) cycling, including uptake and regeneration, in the water column of upper river, lower river, and estuary sites. This 1-year study encompassed periods of flood and drought, in a coastal catchment of south Texas. Low NH4+ concentrations and frequently balanced net NH4+ fluxes suggest minimal N cycling, but these measurements alone did not reveal the patterns of water column NH4+ uptake and regeneration in the river and estuarine systems. Rapid turnover of NH4+ supported productive estuaries, particularly during periods of droughts when riverine sources of NH4+ were minimal. However, NH4+ demand declined during storms across the river–estuary continuum, and regeneration rates were high, especially in the rivers. Most research in rivers has focused on benthic or whole-system dynamics, but our data demonstrate that active NH4+ cycling also occurs in isolated river water columns. Lower river sites were hotspots of NH4+ cycling on the landscape. Continued studies across river–estuary continuums are needed to enhance our understanding of aquatic systems and improve our ability to manage nutrients in the face of increased anthropogenic pressures and a changing climate.

Community Biological Ammonium Demand: A Conceptual Model for Cyanobacteria Blooms in Eutrophic Lakes

Cyanobacterial harmful algal blooms (CyanoHABs) are enhanced by anthropogenic pressures, including excessive nutrient (nitrogen, N, and phosphorus, P) inputs and a warming climate. Severe eutrophication in aquatic systems is often manifested as non-N2-fixing CyanoHABs (e.g., Microcystis spp.), but the biogeochemical relationship between N inputs/dynamics and CyanoHABs needs definition. Community biological ammonium (NH4+) demand (CBAD) relates N dynamics to total microbial productivity and NH4+ deprivation in aquatic systems. A mechanistic conceptual model was constructed by combining nutrient cycling and CBAD observations from a spectrum of lakes to assess N cycling interactions with CyanoHABs. Model predictions were supported with CBAD data from a Microcystis bloom in Maumee Bay, Lake Erie, during summer 2015. Nitrogen compounds are transformed to reduced, more bioavailable forms (e.g., NH4+ and urea) favored by CyanoHABs. During blooms, algal biomass increases faster than internal NH4+ regeneration rates, causing high CBAD values. High turnover rates from cell death and remineralization of labile organic matter consume oxygen and enhance denitrification. These processes drive eutrophic systems to NH4+ limitation or colimitation under warm, shallow conditions and support the need for dual nutrient (N and P) control.

Effects of diurnal vertical mixing and stratification on phytoplankton productivity in geothermal Lake Rotowhero, New Zealand

Abstract Mixing processes in lakes are key factors controlling light availability for phytoplankton growth, but understanding the contribution of mixing is often confounded by other factors such as nutrient availability and species dynamics. Our study examined this problem in a low pH, geothermally heated lake dominated by one phytoplankton genus and lacking the complexity of nutrient limitation, phytoplankton species interactions, or grazing pressure. We hypothesized that the continuous strong convectively driven circulation resulting from atmospheric instability and sediment heating would negate any tendency of thermal stratification, entraining phytoplankton and transporting them away from high surface irradiance that could induce photoinhibition. During our study, water temperatures were considerably warmer than air temperatures, with a diurnal maximum surface temperature of 37.5 °C and minimum of 35.5 °C. Surface heating induced stratification, with a temperature difference of 1–2 °C evident during the day, but there was sufficient heat loss and mixing during the night to erode the stratification and create isothermal conditions. The vertical entrainment velocity driven by convective circulation was on the order of 0.1 mm s−1, but when there was strong solar heating, phytoplankton within the top 0.5 m of the water column still showed depressed photosynthetic quantum efficiencies, as determined with a Pulse Amplitude Modulated fluorometer (PHYTOPAM); however, this depression was less than for phytoplankton cells maintained throughout the day in surface waters with bottle incubations. At other times mixing generated by continuous heating and atmospheric instability meant that phytoplankton did not show photoinhibition; therefore, despite the geothermally driven mixing in Rotowhero, the intensity of solar radiation is still the key mechanism determining the stratification response and resultant photoinhibition of the phytoplankton. Lake Rotowhero provides an excellent natural laboratory to examine the relative time scales of mixing and phytoplankton photoinhibition responses because small changes in solar radiation have such marked impacts on the diurnal stratification and radiation experienced by cells located above the diurnal thermocline.

Wastewater influences nitrogen dynamics in a coastal catchment during a prolonged drought

Abstract Ecosystem function measurements can enhance our understanding of nitrogen (N) delivery in coastal catchments across river and estuary ecosystems. Here, we contrast patterns of N cycling and export in two rivers, one heavily influenced by wastewater treatment plants (WWTP), in a coastal catchment of south Texas. We measured N export from both rivers to the estuary over 2 yr that encompass a severe drought, along with detailed mechanisms of N cycling in river, tidal river, and two estuary sites during prolonged drought. WWTP nutrient inputs stimulated uptake of N, but denitrification resulting in permanent N removal accounted for only a small proportion of total uptake. During drought periods, WWTP N was the primary source of exported N to the estuary, minimizing the influence of episodic storm‐derived nutrients from the WWTP‐influenced river to the estuary. In the site without WWTP influence, the river exported very little N during drought, so storm‐derived nutrient pulses were important for delivering N loads to the estuary. Overall, N is processed from river to estuary, but sustained WWTP‐N loads and periodic floods alter the timing of N delivery and N processing. Research that incorporates empirical measurements of N fluxes from river to estuary can inform management needs in the face of multiple anthropogenic stressors such as demand for freshwater and eutrophication.

Contributions of freshwater mussels (Unionidae) to nutrient cycling in an urban river: filtration, recycling, storage, and removal

Consumers contribute to nutrient cycling in aquatic ecosystems by nutrient retention in tissues, metabolic transformations and excretion, and promoting microbial processes that remove nutrients (i.e., nitrogen (N) loss via denitrification). Freshwater mussels (Unionidae) form dense assemblages in rivers, and affect nutrient transformations through feeding, biodeposition, and bioturbation. However, the effects of Unionid mussels on N and phosphorus (P) retention are not commonly measured. We quantified rates of filtration, retention, and biodeposition of carbon (C), N, and P for two Unionid species: Lasmigona complanata and Pyganodon grandis. We used continuous-flow cores with 15N tracers to measure denitrification in sediments alone and with a single individual of each species. We conducted measurements in an urban river near Chicago, IL, USA that is a target for Unionid restoration. Both Unionid species showed high rates of P-specific feeding and retention, but low N-specific rates. This was in agreement with high N:P ratio in the water column. Each species significantly increased denitrification relative to sediment alone. 15N tracers suggested direct denitrification of nitrate increased denitrification, although enhanced coupled nitrification–denitrification likely also contributed to denitrification. Finally, denitrification rates with and without mussels were used to estimate the value of N loss under different scenarios for mussel restoration at the river scale. Overall, restored Unionid populations may enhance P retention in soft tissues and shells and N loss via denitrification. Ecosystem managers may find greater support for restoration of Unionid populations with careful calculations of their ecosystem role in nutrient retention and removal.