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A comprehensive carbon budget was constructed to quantify carbon flows through the freshwater-marine continuum of a temperate, microtidal estuary. We performed coordinated measurements of dissolved inorganic carbon and total organic carbon fluxes to resolve spatial variability between and along the channel and shoals and diel variability across the entire estuary for 2 yr. Net ecosystem metabolism (NEM) was the most significant control on carbon flow within estuary regions. However, metabolic rates were spatially coupled such that counteracting fluxes across the channel-shoal gradient or along the river-ocean gradient resulted in system-wide NEM that was closely in balance (–3.0 ± 3.3 to 1.1 ± 4.4 molC m−2 yr−1). Similarly, large diel and seasonal variability in air–water CO2 fluxes were observed during 72 spatial surveys, but these short-term variations generally cancelled out when aggregated to annual budget terms. Although atmospheric exchanges were small (–0.2 ± 0.1 to 2.0 ± 0.4 molC m−2 yr−1), they were subject to large errors (± 4 molC m−2 yr−1) if diel variability was neglected. Internal mechanisms that maintained balanced carbon flows were strongly impacted by river discharge and were only apparent by separately quantifying channel and shoal fluxes. Notably, metabolic responses of the shoal to river forcing outweighed the responses of the channel, and the net impact was contrary to prior relationships derived from synthesis of lower-resolution carbon budgets. Our budget demonstrates that resolution of carbon fluxes at appropriate scales, including channel-shoal and diel variability, is critical to characterizing ecosystem function and the fate of carbon within the river-ocean continuum.

Biotic interactions can structure ecological communities and influence ecosystem functioning. As ecosystem engineers and filter feeders, bivalves often have disproportionately large effects on ecosystem functioning. They also utilize numerous morphological and behavioral responses to reduce predation, which can include changes in their filtration rates. To test the response of Crassostrea virginica filtration rates to the presence of predators, juvenile and adult oysters were separately exposed to varying types of predation risk from Callinectes sapidus and Panopeus herbstii in outdoor mesocosms. Water column chlorophyll a concentrations and crab behavior were measured over the duration of the experiment. Predation risk had no effect on oyster reef drawdown of chlorophyll a, which suggests that this important ecosystem function of oyster reefs is not mediated by behaviorally induced predator effects. Therefore, efforts to model how oyster predators influence filtration rates and associated ecosystem services should focus primarily on the factors that influence oyster mortality rather than predator effects on oyster behavior.

We examined seasonal and spatial patterns in dissolved organic carbon (DOC) and chromophoric dissolved organic matter (CDOM) in the Chowan River watershed, North Carolina, a blackwater river which discharges into the second largest estuary in the United States, the Albemarle–Pamlico Estuarine System. From April 2008 to May 2010, DOC concentration did not significantly vary across seasons (range 7.69–30.39 mg L−1); however, CDOM molecular size and aromaticity increased throughout the spring, decreased during the summer and fall, and remained relatively low in the winter. Spectral slope ratios suggested microbial processing of CDOM in the spring and photodegradation of CDOM in the summer and fall. Spatially, DOC and CDOM concentrations were similar in the mainstem and at the mouths of two tributaries, Bennetts Creek and Wiccacon River, but were significantly higher upstream on the tributaries. DOC concentration was positively correlated with CDOM absorbance coefficients at 254 and 350 nm; however, these optical proxies explained only ~60 % of the variance. DOC and CDOM absorption loads to the Albemarle Sound ranged from 2.63 × 1010 g year−1 and 9.84 × 1010 m2 year−1, respectively, in a dry year and 7.9 × 1010 g year−1 and 2.2 × 1011 m2 year−1, respectively, in a wet year, which are comparable to non-blackwater rivers with larger watersheds. Blackwater rivers may therefore represent “hotspots” in coastal carbon chemistry, with seasonal variations in the quality and quantity of DOC and CDOM influencing estuarine food web dynamics and net ecosystem metabolism.

Physical-biological coupling helps structure aquatic communities, yet physical factors can vary widely across large, biogeographic scales. The eastern oyster (Crassostrea virginica) is an ecosystem engineer that creates intertidal reefs, filters water, promotes denitrification, stabilizes shorelines, and provides habitat throughout the inner waters of the U.S. South Atlantic Bight (SAB). We quantified physical variables (temperature, salinity, duration and depth of water inundation), oyster reef properties (slope, vertical relief), and oyster recruitment, density, and biomass over a 1500 km scale across the SAB for one year. All oyster-level and many reef-level variables exhibited unimodal patterns with latitude that peaked in Georgia and South Carolina estuaries. Of the physical variables, salinity and duration of water inundation over reefs were similar across all sites, and temperature declined linearly with increasing latitude, except during summer when it had no relationship with latitude. Depth of water inundation over reefs was the only physical variable with a prominent unimodal distribution that may explain the oyster’s biological responses. Similar durations of water inundation across all reefs coupled with higher water depths in the mid-latitude sites collectively indicate that these sites experience higher flow velocity, energy and net water volume delivery per unit time. The resultant higher accumulation of oyster biomass and heightened reef structure in areas of higher tidal energy emphasize that the physical forcing of the SAB (especially large cross-shelf gradients in tidal amplification) affects the biology of the eastern oyster, including its reef properties, with potential implications for community structure and ecosystem service delivery across a biogeographic scale.

Enhancingpopulations ofsuspension feedingbivalves,particularlythe easternoyster,Crassostreavirginica, has been proposed as a means of mitigating eutrophication in coastal waters. Review of studies evaluating the effects of C. virginica on nitrogen (N) cycling found that oysters can have effects on water quality that vary by orders of magnitude among sites, seasons, and growing condition (e.g., oyster reefs, aquaculture). Nitrogen contained in phytoplankton consumed by oysters may be returned to the water column, assimilatedintooystertissue and shell,buriedin the sediments,orreturnedtothe atmosphereas dinitrogen gas, primarily via denitrification. Accurately quantifying oyster-related N removal requires detailed knowledge oftheseprimaryfatesofNincoastalwaters.Areviewofexistingdatademonstratedthatthecurrentstateof knowledgeisincompleteinmanyrespects.Nitrogenassimilatedintooystertissueandshellpergramofdry weight was generallysimilaracrosssitesand in oysters growingon reefscomparedto aquaculture.Data on long-term burial of N associated with oyster reefs or aquaculture are lacking. When compared to suitable reference sites, denitrification rates were not consistently enhanced. Depending on environmental and oyster growing conditions, changes in denitrification rates varied by orders of magnitude among studies and did not always occur. Oyster aquaculture rarely enhanced denitrification. Unharvested oyster reefs frequently enhanced denitrification rates. Incorporating oysters into nutrient reduction strategies will require filling gaps in existing data to determine the extent to which relationships between N removal and environmental and/or growing conditions can be generalized.

Multiple stressors affect estuarine shorelines including erosion, sea level rise and impacts from human development of adjacent lands. Increasingly common features of coastal development are vertical shoreline stabilization structures such as bulkheads. Bulkheads are designed to prevent land loss and flooding through the construction of a vertical wall anchored to the land. However, they break the connection between land and water and are barriers to upland plant migration. This disconnect can affect hydrology, alter nutrient and sediment supplies, and lead to marsh loss. We measured the effects of bulkheads on sediment nitrogen fluxes, including denitrification (DEN), at three representative estuarine shoreline types: natural marsh (no bulkhead), bulkhead without marsh, and bulkheads with marshes of varying widths. Sediment cores were taken mid-marsh or, 2 m seaward of bulkhead in sites lacking marsh in northern, central and southern coastal regions of North Carolina. Concentrations of N2 and O2 were measured using a membrane inlet mass spectrometer. In addition, sediment organic matter and inorganic nitrogen concentrations were quantified. Average DEN rate was 93.1 ± 7.0 µmol N m−2 h−1 with the highest rates in the summer and lowest rates in the winter. Sediment oxygen demand was positively correlated with DEN rate (R2 = 0.43, p

We hypothesized that mats of a nonnative macroalga, Gracilaria vermiculophylla, which is often found incorporated several centimeters into intertidal mudflat sediments, would increase net denitrification rates relative to bare sediments. At moderate densities (, 40 g dry weight [dry wt] m22), net denitrification rates in June (182.37 6 16.87 mmol N-N2 m22 h21), July (213.19 6 16.30 mmol N-N2 m22 h21), and September (124.82 6 11.17 mmol N-N2 m22 h21) were higher than rates previously documented with macroalgal mats. Compared with rates from bare sediment in June (25.48 6 15.09 mmol N-N2 m22 h21) and September (46.47 6 15.79 mmol N-N2 m22 h21), net denitrification was significantly higher when G. vermiculophylla was present. Rates measured on bare sediment in July (254.81 6 19.86 mmol N-N2 m22 h21) were not significantly different from G. vermiculophylla counterparts, most likely because of highly reduced conditions in G. vermiculophylla cores, which could have limited nitrification. July incubations also demonstrate that at higher densities (, 120 g dry wt m22 G. vermiculophylla), denitrification rates can drop, suggesting a potential biomass threshold for macroalgal enhancement of denitrification.

In shallow photic systems, the benthic filter, including microphytobenthos and denitrifiers, is important in preventing or reducing release of remineralized NH4 + to the water column. Its effectiveness can be impacted by climate-related drivers, including temperature and storminess, which by increasing wind and freshwater delivery can resuspend sediment, reduce salinity and deliver nutrients, total suspended solids, and chromophoric dissolved organic matter (CDOM) to coastal systems. Increases in temperature and freshwater delivery may initiate a cascade of responses affecting benthic metabolism with impacts on sediment properties, which in turn regulate nitrogen cycling processes that either sequester (via microphytobenthos), remove (via denitrification), or increase sediment nitrogen (via remineralization, nitrogen fixation, and dissimilatory nitrate reduction to ammonium). We conducted a seasonal study at shallow stations to assess the effects of freshwater inflow, temperature, wind, light, and CDOM on sediment properties, benthic metabolism, nitrogen cycling processes, and the effectiveness of the benthic filter. We also conducted a depth study to constrain seasonally varying parameters such as temperature to better assess the effects of light availability and water depth on benthic processes. Based on relationships observed between climatic drivers and response variables, we predict a reduction in the effectiveness of the benthic filter over the long term with feedbacks that will increase effluxes of N to the water column with the potential to contribute to system eutrophication. This may push shallow systems past a tipping point where trophic status moves from net autotrophy toward net heterotrophy, with new baselines characterized by degraded water quality.

Assessing nitrogen dynamics in the estuarine landscape is challenging given the unique effects of individual habitats on nitrogen dynamics. We measured net N2 fluxes, sediment oxygen demand, and fluxes of ammonium and nitrate seasonally from five major estuarine habitats: salt marshes, seagrass beds (SAV), oyster reefs, and intertidal and subtidal flats. Net N2 fluxes ranged from 332 ± 116 μmol N-N2 m−2 h−1 from oyster reef sediments in the summer to −67 ± 4 μmol N-N2 m−2 h−1 from SAV in the winter. Oyster reef sediments had the highest rate of N2 production of all habitats. Dissimilatory nitrate reduction to ammonium (DNRA) was measured during the summer and winter. DNRA was low during the winter and ranged from 4.5 ± 3.0 in subtidal flats to 104 ± 34 μmol 15NH 4 + m−2 h−1 in oyster reefs during the summer. Annual denitrification, accounting for seasonal differences in inundation and light, ranged from 161.1 ± 19.2 mmol N-N2 m−2 year−1 for marsh sediments to 509.9 ± 122.7 mmol N-N2 m−2 year−1 for SAV sediments. Given the current habitat distribution in our study system, an estimated 28.3 × 106 mol of N are removed per year or 76 % of estimated watershed nitrogen load. These results indicate that changes in the area and distribution of habitats in the estuarine landscape will impact ecosystem function and services.

Summary 1. Coastal rivers can have long tidally influenced reaches that are affected by tides but do not contain saline water. These tidal freshwater reaches have steep geomorphic gradients where the river transitions from narrow, heavily shaded streams to wide, unshaded channels. The influence of these gradients on river ecosystem production is poorly understood. 2. We characterised gradients in irradiance, geomorphology, water clarity and chlorophyll a along 9- to 16-km tidal freshwater reaches of the Newport and White Oak Rivers in North Carolina, USA, and examined the effect of nutrient enrichment on phytoplankton growth in the Newport River. Underwater irradiance was modelled at 2–4 week intervals along both rivers using measurements of the above-canopy irradiance, canopy cover, water column light attenuation (Kd) and water depth. Suspended material (TSS), dissolved organic carbon (DOC) and chlorophyll a were sampled at 2-week interval at five sites on the Newport River and on four dates at four sites on the White Oak River over the course of one year. 3. Phytoplankton nutrient limitation was assessed at three locations along the tidal gradient. River water was collected during March, April, June and October, and incubated in 10-L plastic outdoor containers under ambient water temperature and sunlight. Additions of inorganic nitrogen and phosphorus served as treatments; growth rate during the 4 days of incubation was calculated from the change in chlorophyll a concentration over time. 4. Canopy cover decreased from more than 90% to

This research details changes in lake algal community structure that occurred during dystrophication. We conducted a paleolimnological investigation of Pungo Lake, a shallow, dystrophic system near the coast of North Carolina, USA. Multiple chemical and biological proxies were measured on a sediment core, including sedimentary photosynthetic pigments, lignin-phenols, nutrients, and δ13C. Data analysis identified three zones of algal community structure corresponding to three regimes of organic matter inputs. Predystrophic conditions represented a period of low organic inputs but substantial algal abundance (diatoms and other algal types). The period of dystrophication preceded European settlement (1850) and showed an increase in organic matter deposition, lignin, and a change in lignin type. Lignin-phenols and δ13C signatures of organic matter indicated that terrestrial organic matter inputs increased during this period, possibly as a result of wetland expansion. Dystrophication also corresponded to an increase in algal groups that favor low light environments (cyanobacteria and cryptophytes).

Tidal freshwater wetlands (TFW) alter nitrogen concentrations in river water, but the role of these processes on a river’s downstream nitrogen delivery is poorly understood. We examined spatial and temporal patterns in denitrification in TFW of four rivers in North Carolina, USA and evaluated the relative importance of denitrification rate and inundation on ecosystem-scale N2 efflux. An empirical model of TFW denitrification was developed to predict N2 efflux using a digital topographic model of the TFW, a time series of water level measurements, and a range of denitrification rates. Additionally, a magnitude-frequency analysis was performed to investigate the relative importance of storm events on decadal patterns in N2 efflux. Spatially, inundation patterns exerted more influence on N2 efflux than did the range of denitrification rate used. Temporal variability in N2 efflux was greatest in the lower half of the tidal rivers (near the saline estuary) where inundation dynamics exerted more influence on N2 efflux than denitrification rate. N2 efflux was highest in the upper half of the rivers following storm runoff, and under these conditions variation in denitrification rate had a larger effect on N2 efflux than variability in inundation. The frequency-magnitude analysis predicted that most N2 efflux occurred during low flow periods when tidal dynamics, not storm events, affected TFW inundation. Tidal hydrology and riparian topography are as important as denitrification rate in calculating nitrogen loss in TFW; we present a simple empirical model that links nitrogen transport in rivers with loss due to denitrification in TFW.

The management, restoration, and conservation of estuarine habitats would benefit from knowledge of habitat-specific functions that reflect important ecosystem services. Secondary production may provide a comprehensive metric of food web support because it synthesizes contributions of local primary production, food subsidies from other habitats, and the protective influences of habitat structure. Despite widespread perceptions of how habitats compare in food web contribution, few methodologically comparable studies on secondary production across multiple estuarine habitats exist. At field sites in North Carolina, USA, annual secondary produc- tion was estimated for macrobenthic infaunal and epifaunal communities in salt marshes, seagrass meadows, oyster reefs, intertidal and subtidal flats, and on shoreline stabilization structures. Habi- tats with hard emergent or biogenic structure generally exhibited higher secondary production than habitats lacking structure. Oyster reef had the highest secondary production, ranging from 467.3 to 853.7 g ash free dry mass (AFDM) m �2 yr �1 , while shoreline stabilization structures ranked high because of dense epifaunal communities. Estimates of secondary production suggest ranking of natural habitats as oyster reef > salt marsh > seagrass > intertidal flat and subtidal flat. Undesir- able impacts of shoreline stabilization structures on adjacent habitats made their inclusion in this ranking of food web support by habitat difficult. The importance of suspension feeders on oyster reefs, shoreline stabilization structures, and in some marshes suggests that secondary production patterns are partly influenced by external subsidies facilitated by support from habitat structure. Consequently, estuarine rehabilitation should include structural habitat elements that will con- tribute to ecosystem production at higher trophic levels. Without such habitat restoration, the fate of estuaries in the USA affected by anthropogenic stressors may be loss of habitat diversity and prevalence of low-trophic-supporting habitats.

Benthic microalgal communities are important components of estuarine food webs and make substantial contributions to coastal materials cycling. Nitrogen is generally the limiting factor for marine primary production; however other factors can limit benthic primary producers because of their access to the additional nutrients found in sediment porewater. Field and laboratory experiments were conducted to test the hypothesis that water column nitrogen supply affects estuarine sandflat benthic microalgal community structure and function. Our field and mesocosm experiments assessed changes at both the population and functional group levels. Simulated water column nitrogen additions increased maximum community photosynthesis in most cases (Pb max from photosynthesis vs. irradiance curves). Additional changes that resulted from nitrogen additions were decreases in porewater phosphate, increases in porewater ammonium, shifts in community composition from N 2 fixing cyanobacteria toward diatoms, and detectable, though not statistically significant increases in biomass (as chlorophyll a ). Results from field and laboratory experiments were quite similar, suggesting that laboratory experiments support accurate predictions of the response of intertidal benthic microalgae to changes in water column nutrient conditions.

SUMMARY 1. Palaeolimnological data were used to investigate drivers of the community of primary producers in Lake Mattamuskeet, North Carolina, U.S.A. This is a large, shallow lake with two basins currently dominated by phytoplankton and macrophytes. The two basins were divided in 1940 by the building of a roadway across the lake, which also corresponded with the divergence in their ecosystem state. 2. Photosynthetic pigments, organic matter and nutrients (P, N, C, S) were analysed in sediment cores from each basin to reconstruct the primary producer community over the past c. 100 years. We sought to answer two questions. First, what changes to the ecosystem resulting from the building of the roadway caused the development of different primary producer communities in the two basins? Second, why have the alternative ecosystem states persisted despite a variety of human perturbations since 1940? 3. K-means cluster analysis and principal component analysis were applied to identify three sediment types based on photosynthetic pigment data: sediments indicating low productivity (low pigment concentrations), sediments associated with macrophytes (chlorophyll a and b) and with phytoplankton (alloxanthin and aphanizophyll). In addition, other palaeolimnological proxies measured, such as loss on ignition, total phosphorus, total organic carbon ⁄ total nitrogen and other nutrients, were different in post1940 sediments within the two basins. 4. These differences suggest characteristics, such as nutrient cycling, water depth and other physical changes resulting from roadway construction, combined to establish and maintain the differing communities of primary producers in the two basins. Furthermore, Fe ⁄ S dynamics and waterfowl herbivory probably contributed to the development of the two ecosystem states.

Lake Mattamuskeet, North Carolina, USA is a large (162 km2) and shallow (mean depth = 1 m) coastal lake, which was significantly modified to support agricultural activities following European settlement in 1850. Paleolimnological proxies measured on a 400-cm sediment core collected from Lake Mattamuskeet reveal shifts in organic matter input and primary producer community structure in response to climatic and human impacts on the lake during the late Holocene. Stratigraphic changes in organic matter content, nutrients, metals, lignin phenols and photosynthetic pigments were used to divide the sediment core into three intervals. Interval I includes sediment deposited between A.D. 360–1584 and indicates a clear-water, sand-bottom state with low algal abundance. In addition, the lake catchment area experienced two significant fires during this interval that were recorded as charcoal layers in the core around A.D. 360 and A.D. 1435 (calibrated 14C AMS dates). Trophic structure changed with the onset of Interval II (A.D. 1584–1860) when total algal abundance increased, and the primary producer community was comprised primarily of diatoms, chrysophytes, cryptophytes and cyanobacteria. During this interval there was also an increase in terrestrial organic material input into the lake as well as a shift in plant type from woody gymnosperms to non-woody angiosperms as determined from lignin data. Sediment deposited in Lake Mattamuskeet following European settlement (Interval III, A.D. 1860-present) suggests a dramatic increase in organic-matter deposition, metals, primary-producer abundance and the onset of cyanobacterial dominance. Sedimentary evidence indicates that shallow-water primary producers can respond rapidly to climate change and human development.

Since the mid 1990s, the Atlantic and Gulf Coast regions have experienced a dramatic increase in the number of hurricane landfalls. In eastern North Carolina alone, eight hurricanes have affected the coast in the past 9 years. These storms have exhibited individualistic hydrologic, nutrient, and sediment loading effects and represent a formidable challenge to nutrient management aimed at reducing eutrophication in the Pamlico Sound and its estuarine tributaries. Different rainfall amounts among hurricanes lead to variable freshwater and nutrient discharge and variable nutrient, organic matter, and sediment enrichment. These enrichments differentially affected physical and chemical properties (salinity, water residence time, transparency, stratification, dissolved oxygen), phytoplankton primary production, and phytoplankton community composition. Contrasting ecological responses were accompanied by changes in nutrient and oxygen cycling, habitat, and higher trophic levels, including different direct effects on fish populations. Floodwaters from the two largest hurricanes, Fran (1996) and Floyd (1999), exerted multi-month to multi-annual effects on hydrology, nutrient loads, productivity, and biotic composition. Relatively low rainfall coastal hurricanes like Isabel (2003) and Ophelia (2005) caused strong vertical mixing and storm surges, but relatively minor hydrologic and nutrient effects. Both hydrologic loading and wind forcing are important drivers and must be integrated with nutrient loading in assessing short-term and long-term ecological effects of these storms. These climatic forcings cannot be managed but should be considered in the development of water quality management strategies for these and other large estuarine ecosystems faced with increasing frequencies and intensities of hurricane activity.

Phytoplankton diagnostic photopigments in near-surface waters (≈0.5 m) were identified and quantified by high-performance liquid chromatography beginning in April 1994 in the Neuse River Estuary and in October 1999 in the Pamlico Sound, North Carolina. Photopigment concentrations were analyzed using ChemTax to determine the class-specific biomass of the dominant phytoplankton groups. Long-term annual and seasonal trends in phytoplankton biomass and composition were characterized along the river-estuarine continuum and compared to river flow rates, which were variable because of droughts, uncharacteristic seasonal rainfall patterns, and, since 1996, an increase in the frequency of tropical storms and hurricanes. We tested the hypothesis that temporal and spatial patterns of phytoplankton biomass and composition were largely controlled by changes in river flow rate through associated changes in salinity and residence time or through physical transport and advection of phytoplankton classes with river flow along the estuary. Significant interannual, seasonal, and spatial variability in phytoplankton biomass and composition was observed and coincided with variability in river flow rates. The five dominant phytoplankton classes (Chlorophyceae, Cryptophyceae, Cyanobacteria, Bacillariophyceae, and Dinophyceae) were physically displaced downstream during periods of elevated river flow. Dinoflagellates were reduced in abundance during high flow conditions, especially in the upper Neuse River Estuary. The abundance of cyanobacteria was also reduced throughout the system during elevated river flow conditions, although chlorophytes were more abundant. Changes in hydrology can be a useful indicator of seasonal phytoplankton distribution and higher level compositional changes along hydrologic gradients in these and similar systems.

Microbial communities growing on the surfaces of standing dead stems of salt marsh grasses contribute to fixation of atmospheric nitrogen (N2) in these N-limited intertidal systems. Salt marshes in North Carolina, USA, have been shown to exhibit N2 fixation throughout the growth sea- son, but the composition of these diazotrophic communities is poorly known. This study investigated the diversity of the epiphytic N2-fixing microbial community on the surfaces of dead stems (shoots) of the cord grass Spartina alterniflora in a salt marsh in North Carolina. Sequencing of the nifH gene and microscopy showed that a diverse diazotrophic community consisting of at least 8 diazotrophic cyanobacterial taxa, as well as an assemblage of α-, γ-, and δ-proteobacteria, was present. Half of the recovered unique nifH sequences fell into the cluster with anaerobes. N2-fixation rates under natural irradiance ranged from 0.001 to 2.58 nmol C2H4 µg chl a -1 h -1 (0.07 to 206 nmol C2H4 cm -2 h -1 ) and decreased from spring to fall. Nitrogen-fixation rates had a positive relationship with chlorophyll a, suggesting phototrophs contributed to N2 fixation. The dominant diazotrophs in this study differed from epiphytic communities described elsewhere in the southeast United States, suggesting regional differences are present in the diversity of these communities.

Uptake rates of ammonium, nitrate and urea were measured during the spring, summer and autumn (2001) in the eutrophic, nitrogen (N) limited Neuse River Estuary (NRE), North Carolina, USA. Ammonium was the dominant form of N taken up during the study, contributing approximately half of the total measured N uptake throughout the estuary. Nitrate uptake declined significantly with distance downstream comprising 33% of the total uptake in the upper estuary but only 11 and 16% in the middle and lower estuary, respectively. Urea uptake contributed least to the total pool in the upper estuary (16%), but increased in importance in the middle and lower estuary, comprising 45 and 37% of the total N taken up, respectively. The importance of regenerated N for fuelling phytoplankton productivity in the mesohaline sections of the NRE is demonstrated. The contribution of urea to the regenerated N pool suggests that internal regeneration of dissolved organic N may support a large proportion of the phytoplankton primary production and biomass accumulation in the middle and lower NRE. These results suggest that N-budgets based on dissolved inorganic N uptake rates alone will seriously under estimate phytoplankton N uptake.

Human activities in the watersheds of the tributaries of Pamlico Sound (PS) in North Carolina have resulted in increased riverine loading of nutrients. Pamlico Sound is a regionally important aquatic resource, and provides crucial foraging and nursery habitats for Southeast Atlantic fisheries. Changes in phytoplankton community composition that may result from increased frequency and quantity of inorganic nutrient inputs could have negative ecological effects on PS. In this study we conducted a series of nutrient bioassays to assess the relationship between increased inorganic nutrient concentration and phytoplankton community structure and function. Experiments were conducted on the native phytoplankton community of the southwest basin of PS. We utilized nutrient addition treatments and all-but-one nutrient treatments in bioassays. This allowed the comparison of the effect of adding one potentially growth-limiting nutrient (e.g. nitrogen) to adding all potentially limiting nutrients except one (e.g. all except nitrogen). Data from these bioassays indicated that the phytoplankton community in PS is primarily nitrogen (N) limited. Dissolved inorganic N concentrations in PS were relatively stable during this study. The biology of its estuarine tributaries, as has been shown for the Neuse River Estuary (NRE), acted as an effective filter for most of the nutrients transported from upstream. We found stoichiometric predictors of phytoplankton community nutrient limitations to be reliable in some instances, but inaccurate in others. Some taxa-specific responses to nutrient additions were observed, however there were no consistent patterns throughout the experiments. Results indicated that changes in the PS phytoplankton community could result from changes in nutrient regime, and changes may not be consistent across phytoplankton taxonomic groups. Unlike the NRE where pulses of riverine N have significant effects on phytoplankton community structure, the PS phytoplankton community did not appear to be subjected to these periodic N enrichments.

This study employed simulated spills of weathered diesel fuel and measured the initial effects on the intertidal sand flat microphytobenthic (MPB) communities. The goals were to examine the impacts of short-term (hours) and longer-term (days) exposure to petroleum on the native sand flat MPB in coastal North Carolina and to assess recovery of the community following the exposure. We assessed changes in biomass (chlorophyll a ), primary productivity ( 14 C bicarbonate incorporation), photophysiology ( P vs. I curves) and species composition (microscopy) and compared diesel exposed samples to unamended controls. We found that short-term impacts of diesel fuel pollution were confined to primary productivity and photophysiology of sand flat MPB. Short-term effects were only detected at relatively high concentrations that are not common outside of a major spill event. In the longer term, diesel fuel was again found to have effects on primary productivity, but at higher concentrations than would be likely to occur in industrialized coastal areas. However, negative impacts on photophysiology were detected at diesel fuel concentrations slightly above typical ambient conditions in coastal waters in industrialized areas. Biomass as measured by chlorophyll a was not affected by any concentration in the longer-term exposure to diesel fuel. Cell counts in the longer-term experiments found cyanobacteria had larger negative impacts from diesel fuel exposure than did diatoms. The recovery portion of this study showed the sand flat MPB communities were fairly resilient following both additions of diesel fuel. However, photophysiology and cell counts did not return to conditions equivalent to the control. Data from this study indicate that the effects of petroleum pollution on the MPB community of tidal sand flats should be considered alongside effects on other coastal microalgae in ecological and damage assessments.

The purpose of this study was to assess the differences in the abundance and activity of the bacterioplankton at sites with varied boating activity, and to determine the response of the communities to additional petroleum pollution. Three sites, including two marinas and a site on Bogue Sound in coastal North Carolina, were selected for monthly experiments. Seasonal patterns of bacterioplanktonic abundance and diesel fuel biodegradation for each site were examined, and possible correlations with intensity of boating activities were explored. Bacterioplanktonic communities at the three sites in this study were more similar in their structure and biodegradation potential than would have been expected from previous studies. We found no differences in the abundance and biodegradation potential of theses communities at the three sites, and only a slightly elevated number of hydrocarbon degraders at one of the marinas. Patterns of biodegradation were more closely related to total bacterial abundance, rather than number of petroleum hydrocarbon degraders. There was a strong seasonal pattern in bacterial abundance and biodegradation at all three sites. The bacterioplanktonic community at all sites responded similarly to additions of diesel fuel. Bacterial abundance and productivity were both elevated to some extent, and number of petroleum hydrocarbon degraders changed relatively little. These data indicate that marinas may not profoundly affect the bacterioplankton and that coastal waters often have resident bacterial communities capable of degrading petroleum hydrocarbon pollution.

A variety of analyses were used to assess the structure (community composition) and function (assimilation number, nitrogen fixation) of phytoplankton in the Neuse River Estuary (NRE), NC under ambient and modified nutrient concentrations. Dilution bioassays were employed to reduce the concentration of nitrogen (N) or both N and phosphorus (P) and thus compare varied DIN:DIP ratios. Experimental manipulations created conditions that may result from mandated N load reductions to the estuary. We hypothesized that unilateral reduction of N loading to the NRE would increase the activity, abundance and diversity of N2 fixing cyanobacteria. Changes in phytoplankton primary productivity, N2 fixation (nitrogenase activity), genetic potential for N2 fixation (presence of nifH), phytoplankton taxonomic composition (diagnostic photopigment concentration) and abundances of N2 fixing cyanobacteria (microscopy) were determined. Decreasing ambient DIN:DIP ratios in NRE samples resulted in increased rates of N2 fixation when seed populations were present and environmental conditions were amenable. Decreasing the DIN:DIP ratio did not lead to an increase in the abundance or diversity of N2 fixing cyanobacteria. Because N2 fixing cyanobacteria were only actively fixing nitrogen during periods of low riverine N discharge (summer and early autumn), lowering nutrient ratios may not have a major impact on the NRE. However, the maximum potential amount of N from N2 fixation was calculated using rates from this study and was found to be approximately 3% of total riverine loading of N to the NRE. Because N2 fixation occurs farther downstream and later in the year than riverine N loading to the NRE, there is potential for N2 fixation to modify N dynamics. Analyses of the phytoplankton community as a whole in these relatively short term experiments indicated that reduced DIN:DIP may not have a major impact on their structure and function.

As sea level rise and human activities erode coastal wetlands, managers rebuild or preserve wetlands that can perform the ecosystem services of a natural system. One increasingly common mitigation activity is the construction of rock sills in the low marsh zone to stabilize marsh elevation. Sills dramatically alter the physical structure of marshes by changing elevation, adding hard substrate and potentially altering the spatial structure of benthic algal communities in and adjacent to the low marsh. We documented differences in benthic algal abundance at the seaward marsh edge in silled and unsilled marshes in North Carolina. We found that sills were associated with reduced standing stocks of benthic algal primary production and reduced macroalgal taxonomic richness, and this difference was driven primarily by differences in macroalgal abundance. We experimentally tested the effect of macroalgal abundance on cordgrass (Spartina alterniflora) growth in the low zone of an unmanipulated marsh, and found that macroalgal removal had no effect on final cordgrass abundance. Our study suggests that salt marsh management through the construction of sills in low marsh zones impacts benthic primary production in the low marsh zone, but that benthic algal production does not affect cordgrass growth over a growing season.