Melody J. Bernot

Stream denitrification across biomes and its response to anthropogenic nitrate loading

Anthropogenic addition of bioavailable nitrogen to the biosphere is increasing and terrestrial ecosystems are becoming increasingly nitrogen-saturated, causing more bioavailable nitrogen to enter groundwater and surface waters. Large-scale nitrogen budgets show that an average of about 20–25 per cent of the nitrogen added to the biosphere is exported from rivers to the ocean or inland basins, indicating that substantial sinks for nitrogen must exist in the landscape. Streams and rivers may themselves be important sinks for bioavailable nitrogen owing to their hydrological connections with terrestrial systems, high rates of biological activity, and streambed sediment environments that favour microbial denitrification. Here we present data from nitrogen stable isotope tracer experiments across 72 streams and 8 regions representing several biomes. We show that total biotic uptake and denitrification of nitrate increase with stream nitrate concentration, but that the efficiency of biotic uptake and denitrification declines as concentration increases, reducing the proportion of in-stream nitrate that is removed from transport. Our data suggest that the total uptake of nitrate is related to ecosystem photosynthesis and that denitrification is related to ecosystem respiration. In addition, we use a stream network model to demonstrate that excess nitrate in streams elicits a disproportionate increase in the fraction of nitrate that is exported to receiving waters and reduces the relative role of small versus large streams as nitrate sinks.

Nitrous oxide emission from denitrification in stream and river networks

Nitrous oxide (N2O) is a potent greenhouse gas that contributes to climate change and stratospheric ozone destruction. Anthropogenic nitrogen (N) loading to river networks is a potentially important source of N2O via microbial denitrification that converts N to N2O and dinitrogen (N2). The fraction of denitrified N that escapes as N2O rather than N2 (i.e., the N2O yield) is an important determinant of how much N2O is produced by river networks, but little is known about the N2O yield in flowing waters. Here, we present the results of whole-stream 15N-tracer additions conducted in 72 headwater streams draining multiple land-use types across the United States. We found that stream denitrification produces N2O at rates that increase with stream water nitrate (NO3−) concentrations, but that <1% of denitrified N is converted to N2O. Unlike some previous studies, we found no relationship between the N2O yield and stream water NO3−. We suggest that increased stream NO3− loading stimulates denitrification and concomitant N2O production, but does not increase the N2O yield. In our study, most streams were sources of N2O to the atmosphere and the highest emission rates were observed in streams draining urban basins. Using a global river network model, we estimate that microbial N transformations (e.g., denitrification and nitrification) convert at least 0.68 Tg·y−1 of anthropogenic N inputs to N2O in river networks, equivalent to 10% of the global anthropogenic N2O emission rate. This estimate of stream and river N2O emissions is three times greater than estimated by the Intergovernmental Panel on Climate Change.

Nitrogen retention, removal, and saturation in lotic ecosystems

Increased nitrogen (N) loading to lotic ecosystems may cause fundamental changes in the ability of streams and rivers to retain or remove N due to the potential for N saturation. Lotic ecosystems will saturate with sustained increases in the N load, but it is unclear at what point saturation will occur. Rates of N transformation in lotic ecosystems will vary depending on the total N load and whether it is an acute or chronic N load. Nitrogen saturation may not occur with only pulsed or short-term increases in N. Overall, saturation of microbial uptake will occur prior to saturation of denitrification of N and denitrification will become saturated prior to nitrification, exacerbating increases in nitrate concentrations and in N export downstream. The rate of N export to downstream ecosystems will increase proportionally to the N load once saturation occurs. Long term data sets showed that smaller lotic ecosystems have a greater capacity to remove in-stream N loads, relative to larger systems. Thus, denitrification is likely to become less important as a N loss mechanism as the stream size increases. There is a great need for long-term studies of N additions in lotic ecosystems and clear distinctions need to be made between ecosystem responses to short-term or periodic increases in N loading and alterations in ecosystem functions due to chronic N loading.

Patterns of denitrification associated with land use in 9 midwestern headwater streams

Abstract The effects of land use on the relationships among denitrification, NO3−-N, dissolved organic C (DOC), and other environmental parameters were examined in 9 headwater streams (3 each in forested, agricultural, and urban-dominated subwatersheds) in the Kalamazoo River Watershed, Michigan. Sediment denitrification rates were determined using the chloramphenicol-amended acetylene inhibition technique. Agricultural streams had high concentrations of NO3−-N, DOC, soluble reactive P, and NH4+-N, whereas forested streams had the lowest concentrations of these nutrients, and urban streams generally exhibited intermediate concentrations. Sediment denitrification rates were highest in agricultural streams and lowest in forested streams throughout the study period. Availability of NO3−-N was the dominant environmental predictor of sediment denitrification rates, limiting denitrification when NO3−-N concentrations were below a calculated threshold of 0.4 mg NO3−-N/L. Other potential controlling variables (e.g., DOC, dissolved O2, water temperature, and sediment organic matter content) influenced denitrification rates secondarily. Despite higher denitrification rates in agricultural and urban streams compared to forested streams, sediment denitrification removed a smaller proportion of the stream NO3−-N load in agricultural and urban streams relative to forested streams.

Comparing Denitrification Estimates for a Texas Estuary by Using Acetylene Inhibition and Membrane Inlet Mass Spectrometry

ABSTRACT Characterizing denitrification rates in aquatic ecosystems is essential to understanding how systems may respond to increased nutrient loading. Thus, it is important to ensure the precision and accuracy of the methods employed for measuring denitrification rates. The acetylene (C2H2) inhibition method is a simple technique for estimating denitrification. However, potential problems, such as inhibition of nitrification and incomplete inhibition of nitrous oxide reduction, may influence rate estimates. Recently, membrane inlet mass spectrometry (MIMS) has been used to measure denitrification in aquatic systems. Comparable results were obtained with MIMS and C2H2 inhibition methods when chloramphenicol was added to C2H2 inhibition assay mixtures to inhibit new synthesis of denitrifying enzymes. Dissolved-oxygen profiles indicated that surface layers of sediment cores subjected to the MIMS flowthrough incubation remained oxic whereas cores incubated using the C2H2 inhibition methods did not. Analysis of the microbial assemblages before and after incubations indicated significant changes in the sediment surface populations during the long flowthrough incubation for MIMS analysis but not during the shorter incubation used for the C2H2 inhibition method. However, bacterial community changes were also small in MIMS cores at the oxygen transition zone where denitrification occurs. The C2H2 inhibition method with chloramphenicol addition, conducted over short incubation intervals, provides a cost-effective method for estimating denitrification, and rate estimates are comparable to those obtained by the MIMS method.

Temporal variation of pharmaceuticals in an urban and agriculturally influenced stream

Pharmaceuticals have become ubiquitous in the aquatic environment. Previous studies consistently demonstrate the prevalence of pharmaceuticals in freshwater but we do not yet know how concentrations vary over time within a given system. Two sites in central Indiana with varying land use in the surrounding watershed (suburban and agricultural) were sampled monthly for pharmaceutical concentrations and stream physiochemical parameters. Sediment samples were also collected at each sampling event for measurement of δ(15)N natural abundance and sediment organic content. Across sites and sampling events, twelve pharmaceuticals were detected including acetaminophen, caffeine, carbamazepine, cotinine, N,N-diethyl-meta-toluamide (DEET), gemfibrozil, ibuprofen, sulfadimethoxine, sulfamethazine, sulfamethoxazole, triclosan, and trimethoprim. Sulfathiazole, lincomycin, and tylosin were not detected at either site at any time. The agriculturally-influenced site had comparable pharmaceutical concentrations to the urban-influenced site. In general, pharmaceutical concentrations increased during winter at both sites and decreased during spring and summer. Multiple regression analyses indicated that water column dissolved oxygen, the number of days since precipitation, and solar radiation influenced total pharmaceutical concentration in the urban-influenced site; whereas pH, chlorophyll a concentration, and total amount of rainfall in the previous 10 days influenced total pharmaceutical concentrations in the agriculturally-influenced site. Pharmaceutical concentrations were not correlated with sediment δ(15)N across or within sites. However, sediment in the urban-influenced site had higher mean δ(15)N signatures relative to sediment in the agriculturally-influenced site. These data indicate pharmaceuticals are persistent in aquatic ecosystems influenced by both agricultural and suburban activity. Pharmaceuticals are designed to have a physiological effect; therefore, it is likely that they may also influence aquatic organisms, potentially threatening freshwater ecosystem health.

Detection of pharmaceuticals and personal care products (PPCPs) in near-shore habitats of southern Lake Michigan

Pharmaceuticals and personal care products (PPCPs) have been documented throughout the United States freshwaters but research has focused largely on lotic systems. Because PPCPs are designed to have a physiological effect, it is likely that they may also influence aquatic organisms. Thus, PPCPs may negatively impact aquatic ecosystems. The objectives of this research were to quantify PPCP abundance in near-shore habitats of southern Lake Michigan and identify factors related to PPCP abundance. Stratified sampling was conducted seasonally at four southern Lake Michigan sites. All sites and depths had measurable PPCP concentrations, with mean individual compound concentrations of acetaminophen (5.36 ng/L), caffeine (31.0 ng/L), carbamazepine (2.23 ng/L), cotinine (4.03 ng/L), gemfibrozil (7.03 ng/L), ibuprofen (7.88 ng/L), lincomycin (4.28 ng/L), naproxen (6.32 ng/L), paraxanthine (1,7-dimethylxanthine; 46.2 ng/L), sulfadimethoxine (0.94 ng/L), sulfamerazine (0.92 ng/L), sulfamethazine (0.92 ng/L), sulfamethoxazole (26.0 ng/L), sulfathiazole (0.92 ng/L), triclocarban (5.72 ng/L), trimethoprim (5.15 ng/L), and tylosin (3.75 ng/L). Concentrations of PPCPs varied significantly among sampling times and locations (river mouth vs offshore), with statistical interactions between the main effects of site and time as well as time and location. Concentrations of PPCPs did not differ with site or depth. Temperature, total carbon, total dissolved solids, dissolved oxygen, and ammonium concentrations were related to total pharmaceutical concentrations. These data indicate that PPCPs are ubiquitous and persistent in southern Lake Michigan, potentially posing harmful effects to aquatic organisms.

Delineating the effects of zebra mussels (Dreissena polymorpha) on N transformation rates using laboratory mesocosms

Abstract Zebra mussels might enhance denitrification rates by altering 3 primary controls: O2, NO3–, and labile C availability. We stocked mesocosms with stream sediments and either no zebra mussels (−ZM) or 10,000 mussels/m2 (+ZM) to examine these potential mechanisms. We measured sediment nitrification (nitrapyrin-inhibition technique), denitrification (chloramphenicol-amended acetylene block technique), sediment O2 profiles (microelectrodes), and a suite of water and sediment characteristics weekly for 3 wk. Nitrification (2-way analysis of variance [ANOVA], p < 0.001) and denitrification (2-way ANOVA, p < 0.001) rates were significantly higher in +ZM than in −ZM mesocosms. High-NH4+ waste from zebra mussels increased sediment nitrification rates, which increased NO3– availability for denitrification. Furthermore, coupled nitrification–denitrification was enhanced by a reduction in the sediment depth to anoxia in the presence of zebra mussels, and this reduction allowed both processes to occur in close spatial proximity. Despite increased denitrification rates, we did not measure an increased proportion of N lost via denitrification from the +ZM mesocosms. The presence of zebra mussels can increase NO3– availability through increased nitrification rates, potentially exacerbating eutrophication of invaded waters.

Human and veterinary pharmaceutical abundance and transport in a rural central Indiana stream influenced by confined animal feeding operations (CAFOs)

Previous research has documented the ubiquity of human and veterinary pharmaceuticals and personal care products (PPCPs) in freshwater, though their persistence and transport is relatively unknown. The objective of this study was to quantify the abundance and transport of human and veterinary PPCPs in a rural, central Indiana stream influenced by confined animal feeding operations (CAFOs). Research objectives also aimed to identify mechanisms controlling abundance and transport. PPCP concentrations and stream physicochemical characteristics were measured monthly over one year at multiple sites along a 60 km reach. Overall, human PPCPs were more abundant and measured at higher concentrations than veterinary pharmaceuticals. Veterinary pharmaceutical concentrations (lincomycin, sulfamethazine) were greatest in stream reaches adjacent to CAFOs. No distinct spatial variation was observed for human PPCPs. However, caffeine and paraxanthine had significant temporal variation with higher concentrations in winter. In contrast, DEET had higher concentrations in summer. Pharmaceutical load (μg/s) ranged from<0.005 to 1808 μg/s across sites, sampling events and pharmaceutical compounds with human PPCPs having higher loads relative to veterinary pharmaceuticals. Reach input ranged from net retention (sulfamethazine in August) to 1667 μg/m/d paraxanthine in March. Triclosan had the highest measured mean input into the reach (661 μg/m/d) and sulfamethazine had the lowest mean input (32 μg/m/d). Across measured compounds, input of PPCPs into the reach was two orders of magnitude lower than nitrate-N input (57,000 μg/m/d). Transport metrics indicated acetaminophen and caffeine are transported farther than triclosan though had lower loss velocities (loss relative to abundance). Loss rate of PPCPs was an order of magnitude lower than nitrate-N loss rate. Human PPCPs were more abundant than veterinary pharmaceuticals in this rural watershed influenced by CAFOs. Further, concentrations had significant temporal and spatial variation highlighting differential sources and fates. Thus, mechanisms driving PPCP retention and transport need to be identified to aid management of these emerging contaminants.

The effects of the psychiatric drug carbamazepine on freshwater invertebrate communities and ecosystem dynamics

Freshwater ecosystems are persistently exposed to pharmaceutical pollutants, including carbamazepine. Despite the ubiquity and recalcitrance of carbamazepine, the effects of this pharmaceutical on freshwater ecosystems and communities are unclear. To better understand how carbamazepine influences the invertebrate community and ecosystem dynamics in freshwaters, we conducted a mesocosm experiment utilizing environmentally relevant concentrations of carbamazepine (200 and 2000 ng/L). Mesocosms were populated with four gastropod taxa (Elimia, Physa, Lymnaea and Helisoma), zooplankton, filamentous algae and phytoplankton. After a 31 d experimental duration, structural equation modeling (SEM) was used to relate changes in the community structure and ecosystem dynamics to carbamazepine exposure. Invertebrate diversity increased in the presence of carbamazepine. Additionally, carbamazepine altered the biomass of Helisoma and Elimia, induced a decline in Daphnia pulex abundance and shifted the zooplankton community toward copepod dominance. Lastly, carbamazepine decreased the decomposition of organic matter and indirectly altered primary production and dissolved nutrient concentrations. Changes in the invertebrate community occurred through both direct (i.e., exposure to carbamazepine) and indirect pathways (i.e., changes in food resource availability). These data indicate that carbamazepine may alter freshwater community structure and ecosystem dynamics and could have profound effects on natural systems.