Linking DNRA community structure and activity in a shallow lagoonal estuarine system

Dissimilatory nitrate reduction to ammonium (DNRA) and denitrification are two nitrate respiration pathways in the microbial nitrogen cycle. Diversity and abundance of denitrifying bacteria have been extensively examined in various ecosystems. However, studies on DNRA bacterial diversity are limited, and the linkage between the structure and activity of DNRA communities has yet to be discovered. We examined the composition, diversity, abundance, and activities of DNRA communities at five sites along a salinity gradient in the New River Estuary, North Carolina, USA, a shallow temporal/lagoonal estuarine system. Sediment slurry incubation experiments with N-15-nitrate were conducted to measure potential DNRA rates, while the abundance of DNRA communities was calculated using quantitative PCR of nrfA genes encoding cytochrome C nitrite reductase, commonly found in DNRA bacteria. A pyrosequencing method targeting nrfA genes was developed using an Ion Torrent sequencer to examine the diversity and composition of DNRA communities within the estuarine sediment community. We found higher levels of nrfA gene abundance and DNRA activities in sediments with higher percent organic content. Pyrosequencing analysis of nrfA genes revealed spatial variation of DNRA communities along the salinity gradient of the New River Estuary. Percent abundance of dominant populations was found to have significant influence on overall activities of DNRA communities. Abundance of dominant DNRA bacteria and organic carbon availability are important regulators of DNRA activities in the eutrophic New River Estuary

Dynamic modeling of nitrogen losses in river networks unravels the coupled effects of hydrological and biogeochemical processes

The importance of lotic systems as sinks for nitrogen inputs is well recognized. A fraction of nitrogen in streamflow is removed to the atmosphere via denitrification with the remainder exported in streamflow as nitrogen loads. At the watershed scale, there is a keen interest in understanding the factors that control the fate of nitrogen throughout the stream channel network, with particular attention to the processes that deliver large nitrogen loads to sensitive coastal ecosystems. We use a dynamic stream transport model to assess biogeochemical (nitrate loadings, concentration, temperature) and hydrological (discharge, depth, velocity) effects on reach-scale denitrification and nitrate removal in the river networks of two watersheds having widely differing levels of nitrate enrichment but nearly identical discharges. Stream denitrification is estimated by regression as a nonlinear function of nitrate concentration, streamflow, and temperature, using more than 300 published measurements from a variety of US streams. These relations are used in the stream transport model to characterize nitrate dynamics related to denitrification at a monthly time scale in the stream reaches of the two watersheds. Results indicate that the nitrate removal efficiency of streams, as measured by the percentage of the stream nitrate flux removed via denitrification per unit length of channel, is appreciably reduced during months with high discharge and nitrate flux and increases during months of low-discharge and flux. Biogeochemical factors, including land use, nitrate inputs, and stream concentrations, are a major control on reach-scale denitrification, evidenced by the disproportionately lower nitrate removal efficiency in streams of the highly nitrate-enriched watershed as compared with that in similarly sized streams in the less nitrate-enriched watershed. Sensitivity analyses reveal that these important biogeochemical factors and physical hydrological factors contribute nearly equally to seasonal and stream-size related variations in the percentage of the stream nitrate flux removed in each watershed

Stable isotope monitoring of benthic–planktonic coupling using salt marsh fish

Author Posting. © Inter-Research, 2008. This article is posted here by permission of Inter-Research for personal use, not for redistribution. The definitive version was published in Marine Ecology Progress Series 369 (2008): 193-204, doi:10.3354/meps07644.Salt marshes are important coastal ecosystems whose trophic function can be monitored with stable isotopes of abundant fish biosentinel species such as the mummichog Fundulus heteroclitus and the Atlantic silverside Menidia menidia. We compared movement patterns and feeding biology of these species in the summers of 1999 and 2000 in the Rowley River salt marsh estuary north of Boston, Massachusetts, USA. A 15N tracer addition experiment showed that fish of both species were more resident than transient, with mummichogs resident at scales of 1 km or less. Natural abundance stable isotope C, N, and S distributions showed that mummichogs feed more strongly in the benthic food web while silversides feed more in the planktonic food web, with % benthic feeding respectively averaging 58 ± 5 and 32 ± 3% (mean ± 95% confidence limit, CL). For both species, isotope results indicated considerable individual specialization in foraging behavior, likely related to use of channel habitat versus use of the marsh. Power analysis showed that measuring 3 composite samples each comprising 10 to 15 individual fish should provide relatively low errors of 0.5‰ (95% CL) or less around stable isotope averages. Use of such composite samples in monitoring programs will allow detection of significant temporal and spatial changes in benthic-planktonic coupling for salt marsh ecosystems, as recorded in average fish diets. Analyzing some individual fish also is recommended to obtain more detailed information on fish food sources, feeding specializations, and end-member isotope values used in estimating importance of benthic and planktonic food sources.This work was supported by Louisiana SeaGrant Projects R/CEH-13 and R-EFH-07, NOAA MULTISTRESS award 16OP2670, and NSF award DEB 9815598 and BES SGER award 0553138-001

The importance of dissimilatory nitrate reduction to ammonium (DNRA) in the nitrogen cycle of coastal ecosystems

Until recently, it was believed that biological assimilation and gaseous nitrogen (N) loss through denitrification were the two major fates of nitrate entering or produced within most coastal ecosystems. Denitrification is often viewed as an important ecosystem service that removes reactive N from the ecosystem. However, there is a competing nitrate reduction process, dissimilatory nitrate reduction to ammonium (DNRA), that conserves N within the ecosystem. The recent application of nitrogen stable isotopes as tracers has generated growing evidence that DNRA is a major nitrogen pathway that cannot be ignored. Measurements comparing the importance of denitrification vs. DNRA in 55 coastal sites found that DNRA accounted for more than 30% of the nitrate reduction at 26 sites. DNRA was the dominant pathway at more than one-third of the sites. Understanding what controls the relative importance of denitrification and DNRA, and how the balance changes with increased nitrogen loading, is of critical importance for predicting eutrophication trajectories. Recent improvements in methods for assessing rates of DNRA have helped refine our understanding of the rates and controls of this process, but accurate measurements in vegetated sediment still remain a challenge

Functional polymorphisms in the P2X7 receptor gene are associated with stress fracture injury

Context: Military recruits and elite athletes are susceptible to stress fracture injuries. Genetic predisposition has been postulated to have a role in their development. The P2X7 receptor (P2X7R) gene, a key regulator of bone remodelling, is a genetic candidate that may contribute to stress fracture predisposition. Objective: To evaluate the putative contribution of P2X7R to stress fracture injury in two separate cohorts, military personnel and elite athletes. Methods: In 210 Israeli Defence Forces (IDF) military conscripts, stress fracture injury was diagnosed (n=43) based on symptoms and a positive bone scan. In a separate cohort of 518 elite athletes, self-reported medical imaging scan-certified stress fracture injuries were recorded (n=125). Non-stress fracture controls were identified from these cohorts who had a normal bone scan or no history or symptoms of stress fracture injury. Study participants were genotyped for functional SNPs within the P2X7R gene using proprietary fluorescence-based competitive allele-specific PCR assay. Pearson Chi-square (χ2) tests, corrected for multiple comparisons, were used to assess associations in genotype frequencies. Results: The variant allele of P2X7R SNP rs3751143 (Glu496Ala- loss of function) was associated with stress fracture injury, while the variant allele of rs1718119 (Ala348Thr- gain of function) was associated with a reduced occurrence of stress fracture injury in military conscripts (P<0.05). The association of the variant allele of rs3751143 with stress fractures was replicated in elite athletes (P<0.05), whereas the variant allele of rs1718119 was also associated with reduced multiple stress fracture cases in elite athletes (P<0.05). Conclusions: The association between independent P2X7R polymorphisms with stress fracture prevalence supports the role of a genetic predisposition in the development of stress fracture injury

A Critical Review of the \u3csup\u3e15\u3c/sup\u3eN\u3csub\u3e2\u3c/sub\u3e Tracer Method to Measure Diazotrophic Production in Pelagic Ecosystems

Dinitrogen (N2) fixation is an important source of biologically reactive nitrogen (N) to the global ocean. The magnitude of this flux, however, remains uncertain, in part because N2 fixation rates have been estimated following divergent protocols and because associated levels of uncertainty are seldom reported—confounding comparison and extrapolation of rate measurements. A growing number of reports of relatively low but potentially significant rates of N2 fixation in regions such as oxygen minimum zones, the mesopelagic water column of the tropical and subtropical oceans, and polar waters further highlights the need for standardized methodological protocols for measurements of N2 fixation rates and for calculations of detection limits and propagated error terms. To this end, we examine current protocols of the 15N2 tracer method used for estimating diazotrophic rates, present results of experiments testing the validity of specific practices, and describe established metrics for reporting detection limits. We put forth a set of recommendations for best practices to estimate N2 fixation rates using 15N2 tracer, with the goal of fostering transparency in reporting sources of uncertainty in estimates, and to render N2 fixation rate estimates intercomparable among studies

Hydrogeomorphology of the Hyporheic Zone: Stream Solute and Fine Particle Interactions With a Dynamic Streambed

Hyporheic flow in streams has typically been studied separately from geomorphic processes. We investigated interactions between bed mobility and dynamic hyporheic storage of solutes and fine particles in a sand-bed stream before, during, and after a flood. A conservatively transported solute tracer (bromide) and a fine particles tracer (5 μm latex particles), a surrogate for fine particulate organic matter, were co-injected during base flow. The tracers were differentially stored, with fine particles penetrating more shallowly in hyporheic flow and retained more efficiently due to the high rate of particle filtration in bed sediment compared to solute. Tracer injections lasted 3.5 h after which we released a small flood from an upstream dam one hour later. Due to shallower storage in the bed, fine particles were rapidly entrained during the rising limb of the flood hydrograph. Rather than being flushed by the flood, we observed that solutes were stored longer due to expansion of hyporheic flow paths beneath the temporarily enlarged bedforms. Three important timescales determined the fate of solutes and fine particles: (1) flood duration, (2) relaxation time of flood-enlarged bedforms back to base flow dimensions, and (3) resulting adjustments and lag times of hyporheic flow. Recurrent transitions between these timescales explain why we observed a peak accumulation of natural particulate organic matter between 2 and 4 cm deep in the bed, i.e., below the scour layer of mobile bedforms but above the maximum depth of particle filtration in hyporheic flow paths. Thus, physical interactions between bed mobility and hyporheic transport influence how organic matter is stored in the bed and how long it is retained, which affects decomposition rate and metabolism of this southeastern Coastal Plain stream. In summary we found that dynamic interactions between hyporheic flow, bed mobility, and flow variation had strong but differential influences on base flow retention and flood mobilization of solutes and fine particulates. These hydrogeomorphic relationships have implications for microbial respiration of organic matter, carbon and nutrient cycling, and fate of contaminants in streams