Role of C4 carbon fixation in Ulva prolifera, the macroalga responsible for the world's largest green tides.

Most marine algae preferentially assimilate CO2 via the Calvin-Benson Cycle (C3) and catalyze HCO3- dehydration via carbonic anhydrase (CA) as a CO2-compensatory mechanism, but certain species utilize the Hatch-Slack Cycle (C4) to enhance photosynthesis. The occurrence and importance of the C4 pathway remains uncertain, however. Here, we demonstrate that carbon fixation in Ulva prolifera, a species responsible for massive green tides, involves a combination of C3 and C4 pathways, and a CA-supported HCO3- mechanism. Analysis of CA and key C3 and C4 enzymes, and subsequent analysis of δ13C photosynthetic products showed that the species assimilates CO2 predominately via the C3 pathway, uses HCO3- via the CA mechanism at low CO2 levels, and takes advantage of high irradiance using the C4 pathway. This active and multi-faceted carbon acquisition strategy is advantageous for the formation of massive blooms, as thick floating mats are subject to intense surface irradiance and CO2 limitation.

delta(15)N and delta(13)C in the Mondego estuary food web: seasonal variation in producers and consumers.

Assessments of temporal variation in stable carbon and nitrogen ratios were used to examine seasonal trends of the water column and benthic food webs in the Mondego estuary (Portugal). There was a marked seasonality in weather and water column conditions, including nutrient supply and chlorophyll concentrations. In spite of the pronounced environmental changes, we found little evidence of seasonal variation in delta(13)C and delta(15)N of producers and consumers in the Mondego estuary, with a few notable exceptions. Nitrogen isotope ratios in macrophytes (Zostera noltii, Ulva sp., Enteromorpha sp., and Gracilaria sp.), and in two grazers (Idotea chelipes and Lekanesphaera levii) increased during late summer, with the highest delta(15)N values being measured in July, during a period of elevated temperatures and drought, which may have favored high rates of denitrification and heavier delta(15)N values. The results suggest that stable-isotope values from macrophytes and selected grazers are useful as tracers of seasonal changes in nitrogen inputs into estuaries, and that those of consumers reflect other factors beyond seasonal variations in N and C sources.

Supporting Spartina: Interdisciplinary perspective shows Spartina as a distinct solid genus.

In 2014, a DNA-based phylogenetic study confirming the paraphyly of the grass subtribe Sporobolinae proposed the creation of a large monophyletic genus Sporobolus, including (among others) species previously included in the genera Spartina, Calamovilfa, and Sporobolus. Spartina species have contributed substantially (and continue contributing) to our knowledge in multiple disciplines, including ecology, evolutionary biology, molecular biology, biogeography, experimental ecology, biological invasions, environmental management, restoration ecology, history, economics, and sociology. There is no rationale so compelling to subsume the name Spartina as a subgenus that could rival the striking, global iconic history and use of the name Spartina for over 200 yr. We do not agree with the subjective arguments underlying the proposal to change Spartina to Sporobolus. We understand the importance of both the objective phylogenetic insights and of the subjective formalized nomenclature and hope that by opening this debate we will encourage positive feedback that will strengthen taxonomic decisions with an interdisciplinary perspective. We consider that the strongly distinct, monophyletic clade Spartina should simply and efficiently be treated as the genus Spartina.

Effect of field exposure to 38-year-old residual petroleum hydrocarbons on growth, condition index, and filtration rate of the ribbed mussel, Geukensia demissa.

In September 1969, the Florida barge spilled 700,000 L of No. 2 fuel oil into the salt marsh sediments of Wild Harbor, MA. Today a substantial amount, approximately 100 kg, of moderately degraded petroleum remains within the sediment and along eroding creek banks. The ribbed mussels, Geukensia demissa, which inhabit the salt marsh creek bank, are exposed to the spilled oil. Examination of short-term exposure was done with transplantation of G. demissa from a control site, Great Sippewissett marsh, into Wild Harbor. We also examined the effects of long-term exposure with transplantation of mussels from Wild Harbor into Great Sippewissett. Both the short- and long-term exposure transplants exhibited slower growth rates, shorter mean shell lengths, lower condition indices, and decreased filtration rates. The results add new knowledge about long-term consequences of spilled oil, a dimension that should be included when assessing oil-impacted areas and developing management plans designed to restore, rehabilitate, or replace impacted areas.

Spartina alterniflora δ15N as an indicator of estuarine nitrogen load and sources in Cape Cod estuaries.

δ15N values of coastal biota have been used as indicators of land-derived N-loads and sources to estuarine systems and should respond predictably to differences in nitrogen and be sensitive to changes in nitrogen, preferably at the low end of eutrophication. We evaluated Spartina alterniflora as an indicator species of N-loads and sources of δ15N throughout the growing season, and compared the average δ15N to estuarine nitrogen loads and sources for several estuaries receiving different watershed N-loads. δ15N of S. alterniflora differed among estuaries, and these differences were maintained even as δ15N declined during the end of the growing season. δ15N values increased with increasing nitrogen loads to the subestuaries and with increasing percent wastewater-derived nitrogen load. The response of δ15N of S. alterniflora to increased N loads was greater at low N-loads, and decreased as N-loads increased, suggesting that S. alterniflora is a good indicator of incipient nitrogen load.

Tropical land-sea couplings: Role of watershed deforestation, mangrove estuary processing, and marine inputs on N fluxes in coastal Pacific Panama.

We review data from coastal Pacific Panama and other tropical coasts with two aims. First, we defined inputs and losses of nitrogen (N) mediating connectivity of watersheds, mangrove estuaries, and coastal sea. N entering watersheds-mainly via N fixation (79-86%)-was largely intercepted; N discharges to mangrove estuaries (3-6%), small compared to N inputs to watersheds, nonetheless significantly supplied N to mangrove estuaries. Inputs to mangrove estuaries (including watershed discharges, and marine inputs during flood tides) were matched by losses (mainly denitrification and export during ebb tides). Mangrove estuary subsidies of coastal marine food webs take place by export of forms of N [DON (62.5%), PN (9.1%), and litter N (12.9%)] that provide dissimilative and assimilative subsidies. N fixation, denitrification, and tidal exchanges were major processes, and DON was major form of N involved in connecting fluxes in and out of mangrove estuaries. Second, we assessed effects of watershed forest cover on connectivity. Decreased watershed forest cover lowered N inputs, interception, and discharge into receiving mangrove estuaries. These imprints of forest cover were erased during transit of N through estuaries, owing to internal N cycle transformations, and differences in relative area of watersheds and estuaries. Largest losses of N consisted of water transport of energy-rich compounds, particularly DON. N losses were similar in magnitude to N inputs from sea, calculated without considering contribution by intermittent coastal upwelling, and hence likely under-estimated. Pacific Panama mangrove estuaries are exposed to major inputs of N from land and sea, which emphasizes the high degree of bi-directional connectivity in these coupled ecosystems. Pacific Panama is still lightly affected by human or global changes. Increased deforestation can be expected, as well as changes in ENSO, which will surely raise watershed-derived loads of N, as well as significantly change marine N inputs affecting coastal coupled ecosystems.

Stable isotopic evidence of nitrogen sources and C4 metabolism driving the world's largest macroalgal green tides in the Yellow Sea.

During recent years, rapid seasonal growth of macroalgae covered extensive areas within the Yellow Sea, developing the world's most spatially extensive "green tide". The remarkably fast accumulation of macroalgal biomass is the joint result of high nitrogen supplies in Yellow Sea waters, plus ability of the macroalgae to optionally use C4 photosynthetic pathways that facilitate rapid growth. Stable isotopic evidence shows that the high nitrogen supply is derived from anthropogenic sources, conveyed from watersheds via river discharges, and by direct atmospheric deposition. Wastewater and manures supply about half the nitrogen used by the macroalgae, fertiliser and atmospheric deposition each furnish about a quarter of the nitrogen in macroalgae. The massive green tides affecting the Yellow Sea are likely to increase, with significant current and future environmental and human consequences. Addressing these changing trajectories will demand concerted investment in new basic and applied research as the basis for developing management policies.

Long-term nutrient addition increases respiration and nitrous oxide emissions in a New England salt marsh.

Salt marshes may act either as greenhouse gas (GHG) sources or sinks depending on hydrological conditions, vegetation communities, and nutrient availability. In recent decades, eutrophication has emerged as a major driver of change in salt marsh ecosystems. An ongoing fertilization experiment at the Great Sippewissett Marsh (Cape Cod, USA) allows for observation of the results of over four decades of nutrient addition. Here, nutrient enrichment stimulated changes to vegetation communities that, over time, have resulted in increased elevation of the marsh platform. In this study, we measured fluxes of carbon dioxide (CO 2), methane (CH 4) and nitrous oxide (N2O) in dominant vegetation zones along elevation gradients of chronically fertilized (1,572 kg N ha-1 year-1) and unfertilized (12 kg N ha-1 year-1) experimental plots at Great Sippewissett Marsh. Flux measurements were performed using darkened chambers to focus on community respiration and excluded photosynthetic CO 2 uptake. We hypothesized that N-replete conditions in fertilized plots would result in larger N2O emissions relative to control plots and that higher elevations caused by nutrient enrichment would support increased CO 2 and N2O and decreased CH 4 emissions due to the potential for more oxygen diffusion into sediment. Patterns of GHG emission supported our hypotheses. Fertilized plots were substantially larger sources of N2O and had higher community respiration rates relative to control plots, due to large emissions of these GHGs at higher elevations. While CH 4 emissions displayed a negative relationship with elevation, they were generally small across elevation gradients and nutrient enrichment treatments. Our results demonstrate that at decadal scales, vegetation community shifts and associated elevation changes driven by chronic eutrophication affect GHG emission from salt marshes. Results demonstrate the necessity of long-term fertilization experiments to understand impacts of eutrophication on ecosystem function and have implications for how chronic eutrophication may impact the role that salt marshes play in sequestering C and N.

Transient coastal landscapes: Rising sea level threatens salt marshes.

Salt marshes are important coastal environments that provide key ecological services. As sea level rise has accelerated globally, concerns about the ability of salt marshes to survive submergence are increasing. Previous estimates of likely survival of salt marshes were based on ratios of sea level rise to marsh platform accretion. Here we took advantage of an unusual, long-term (1979-2015), spatially detailed comparison of changes in a representative New England salt marsh to provide an empirical estimate of habitat losses based on actual measurements. We show prominent changes in habitat mosaic within the marsh, consistent and coincident with increased submergence and coastal erosion. Model results suggest that at current rates of sea level rise, marsh platform accretion, habitat loss, and with the limitation of the widespread "coastal squeeze", the entire ecosystem might disappear by the beginning of the next century, a fate that might be likely for many salt marshes elsewhere. Meta-analysis of available data suggests that 40 to 95% of the world's salt marshes will be submerged, depending on whether sea level rise remains at current or reaches anticipated rates for the end of this century.

External and local controls on land-sea coupling assessed by stable isotopic signatures of mangrove producers in estuaries of Pacific Panama.

Foliar stable isotopic signatures of nitrogen, carbon, and sulfur in mangrove vegetation from the Pacific coast of Panama were insensitive to inputs from watersheds with different area of forest land cover, and to seasonal, inter-annual, and global-scale-driven contrasts in rainfall and upwelling. N, C, and S content of mangrove vegetation were not affected by inputs from watersheds with different degrees of deforestation, but showed some influence of down-estuary transformations. While there was substantial variation that remained un-explained, isotopic signatures and nutrient contents were largely determined by species-specific features, and showed substantial small-scale variation reflecting local differences, within-estuary plant-sediment links. The ability of mangrove estuaries to erase effects of deforestation points out that conservation of these wetland ecosystems is important, because, at least in the sites we studied, transformations within mangrove estuaries were strong enough to protect water quality in receiving coastal waters.

The use of sedimentary %C, %N, delta(15)N, and Pb concentrations to assess historical changes in anthropogenic influence on Portuguese estuaries.

Vertical profiles of C, N, delta(15)N and Pb were measured in the Mondego and Mira estuaries as markers that conveyed notions as to the relative influence of anthropogenic influence over the past decades. Recent carbon changes in both estuaries may reflect changes in estuarine productivity, probably as a consequence of sediment reworking and erosion, and also of losses in salt marsh area and dwarf eelgrass beds. delta(15)N values began to diverge considerably before the %C and %N and were higher in Mondego. delta(15)N signatures detected N enrichment at relatively low rates, and indicated that Mondego received more enriched N than Mira. Lead concentrations differed between estuaries, with higher concentrations in Mondego. The secular increase in %N, Pb, and delta(15)N signatures was significantly related to human density in the watersheds of the estuaries and were sensitive indicators of anthropogenic activity.

Long-term biological effects of petroleum residues on fiddler crabs in salt marshes.

In September 1969, the Florida barge spilled 700,000L of No. 2 fuel oil into the salt marsh sediments of Wild Harbor (Buzzards Bay, MA). Today the aboveground environment appears unaffected, but a substantial amount of moderately degraded petroleum still remains 8-20cm below the surface. The salt marsh fiddler crabs, Uca pugnax, burrow into the sediments at depths of 5-25cm, and are chronically exposed to the spilled oil. Behavioral studies conducted with U. pugnax from Wild Harbor and a control site, Great Sippewissett marsh, found that crabs exposed to the oil avoided burrowing into oiled layers, suffered delayed escape responses, lowered feeding rates, and achieved lower densities. The oil residues are therefore biologically active and affect U. pugnax populations. Our results add new knowledge about long-term consequences of spilled oil, a dimension that should be included when assessing oil-impacted areas and developing management plans designed to restore, rehabilitate, or replace impacted areas.

Nitrogen loads to estuaries from waste water plumes: modeling and isotopic approaches.

We developed, and applied in two sites, novel methods to measure ground water-borne nitrogen loads to receiving estuaries from plumes resulting from land disposal of waste water treatment plant (WWTP) effluent. In addition, we quantified nitrogen losses from WWTP effluent during transport through watersheds. WWTP load to receiving water was estimated as the difference between total measured ground water-transported nitrogen load and modeled load from major nitrogen sources other than the WWTP. To test estimated WWTP loads, we applied two additional methods. First, we quantified total annual waste water nitrogen load from watersheds based on nitrogen stable isotopic signatures of primary producers in receiving water. Second, we used published data on ground water nitrogen concentrations in an array of wells to estimate dimensions of the plume and quantify the annual mass of nitrogen transported within the plume. Loss of nitrogen during transport through the watershed was estimated as the difference between the annual mass of nitrogen applied to watersheds as treatment plant effluent and the estimated nitrogen load reaching receiving water. In one plume, we corroborated our estimated nitrogen loss in watersheds using data from multiple-level sampling wells to calculate the loss of nitrogen relative to a conservative tracer. The results suggest that nitrogen from the plumes is discharging to the estuaries but that substantial nitrogen loss occurs during transport through the watersheds. The measured vs. modeled and stable isotopic approaches, in comparison to the plume mapping approach, may more reliably quantify ground water-transported WWTP loads to estuaries.

Eutrophication of Cape Cod estuaries: Effect of decadal changes in global-driven atmospheric and local-scale wastewater nutrient loads.

Nitrogen (N) supply by atmospheric deposition, wastewater, and fertilizers controls estuarine eutrophication. In New England, atmospheric N loads recently decreased by 50% and land-derived contributions rose about 80%, owing to national-scale emission controls and local urban development. The decrease in atmospheric deposition was large enough to balance increases in land-derived N loads, so total N loads to Waquoit Bay estuaries in Cape Cod did not change significantly between 1990 and 2014. Unchanged N regimes were corroborated by finding no differences in estuarine nutrient concentrations and macrophyte biomass between pre-2005 and in 2015. Coastal zones, subject to reasonably rapid changes in global and local driver variables, will require that assessment and management of eutrophication include adaptive strategies that capture effects of changing baselines. Management initiatives will be constrained by spatial scale of driver variables: local efforts may address wastewater and fertilizer N sources, but atmospheric sources require national or international attention.

Isotopic studies in Pacific Panama mangrove estuaries reveal lack of effect of watershed deforestation on food webs.

Stable isotopic N, C, and S in food webs of 8 mangrove estuaries on the Pacific coast of Panama were measured to 1) determine whether the degree of deforestation of tropical forests on the contributing watersheds was detectable within the estuarine food web, and 2) define external sources of the food webs within the mangrove estuaries. Even though terrestrial rain forest cover on the contributing watersheds differed between 23 and 92%, the effect of deforestation was not detectable on stable isotopic values in food webs present at the mouth of the receiving estuaries. We used stable isotopic measures to identify producers or organic sources that supported the estuarine food web. N isotopic values of consumers spanned a broad range, from about 2.7 to 12.3‰. Mean δ(15)N of primary producers and organic matter varied from 3.3 for macroalgae to 4.7‰ for suspended particulate matter and large particulate matter. The δ(13)C consumer data varied between -26 and -9‰, but isotopic values of the major apparent producers or organic matter sampled could not account for this range variability. The structure of the food web was clarified when we added literature isotopic values of microphytobenthos and coralline algae, suggesting that these, or other producers with similar isotopic signature, may be part of the food webs.

Disturbance of salt marsh vegetation by wrack mats in Great Sippewissett Marsh.

To quantify disturbance to salt marsh vegetation, and to test the notion that disturbance and species richness are related, we studied disturbance of vegetation by 195 wrack mats that had become stranded over Great Sippewissett Marsh. The mats varied in area, thickness, residence time, and elevation of stranding. Mats that were large, that had residence times of 3-4 months, and were stranded at higher elevations produced the most damage, ranging from total eradication to a decrease in shoot density of underlying vegatation. Thickness had no effect. About 70% of mats caused no damage, and of those that did, damage was usually 50% or less of the area beneath the mat. Plant species were differentially distributed along the tidal excursion within the marsh, and there were more species (8-9) high in the intertidal range than lower (4-6 species). Vertical distribution of species, species richness, and rate of annual disturbance were not well correlated, although highest and lowest disturbance rates corresponded to highest and lowest species richness. Most of the increases in species richness, however, occurred over a very small range of disturbance, so narrow that it seemed unlikely that disturbance affected richness. It seems more likely that factors other than disturbance rate control vertical position of species in salt marshes. It also appears that the effect of disturbance by wrack is less important than has been claimed.

The role of phenolic compounds and other plant constituents in feeding by Canada geese in a coastal marsh.

We have assessed the relative importance of phenolic compounds, other secondary metabolites, and gross nutrient levels as feeding cues to Canada geese. Phenolic content was the most significant constituent influencing feeding selection by geese. Nutrient content had little or no effect on feeding selection.Correlative data showing the negative influence of plant phenolics on food choices by wild geese were supported by feeding preference tests. Extracts of unpalatable plants inhibited feeding by captive geese relative to extracts of palatable plants. In high phenolic plants, the phenolic containing methanol extract was more inhibitory than extractions made with petroleum ether. In a relatively low phenolic, unpalatable plant, an inhibitory factor was extractable in petroleum ether, indicating that for this species, another class of deterrents was involved. Preference tests with individual secondary metabolites showed that tannic acid and quebracho tannin were very effective in inhibiting feeding by geese and phenolic acids were slightly inhibitory, but a sesquiterpene lactone was not deterrent. These results point out the primacy of some secondary metabolites in determining food choices by geese.

Nitrogen, phenolic acids, and other feeding cues for salt marsh detritivores.

A number of feeding cues determine the palatability of detritus to detritivorous invertebrates. In salt marsh detritus the feeding cues include flavor provided by phenolics such as ferulic acid, pH, salinity, and nitrogen content. We examined the feeding responses to each cue by using experimental manipulations where we changed concentrations of these chemical cues. The palatability of detritus of the salt marsh grass Spartina alterniflora to the detritivorous snail Melampus bidentatus is reduced by increases in ferulic acid in the detritus. The acidity of the acid is partly responsible for inhibition of feeding, but other flavors of the ferulic acid are the major factor. Changes in salinity makes detritus more or less palatable to different species of detritivores. Available nitrogen confers greater palatability to detritus.In the field the feeding cues are all present simultaneously, and detritivores feed based on a hierarchy of cues. For M. bidentatus the presence of sufficient available nitrogen overwhelms the response to ferulic acid, the second most important cue. Salinity and pH, although used as discriminatory cues by themselves, are located lower in the hierarchy than nitrogen and ferulic acid.As detritus ages in the marsh, nitrogen content changes. Similarly, detritus from eutrophied environments shows enhanced nitrogen content. The differences in palatability of new and old detritus, and from eutrophic and non-eutrophic environments, suggests that detritivores respond not to total nitrogen but rather to the availabel nitrogen fractions. The increase in lignin found in old and eutrophic detritus may increase the binding of available nitrogen, and hence reduce the palatability of detritus.

Nitrous oxide sources and sinks in coastal aquifers and coupled estuarine receiving waters.

Sources and sinks of the atmospherically reactive gas nitrous oxide (N(2)O) were determined in the heavily nutrient loaded Childs River in Cape Cod, MA. Surface waters were supersaturated and bottom waters were depleted with N(2)O throughout the system. In apparent septic effluent plumes, N(2)O concentrations reached 3 orders of magnitude above atmospheric equilibrium. Because nitrate and N(2)O concentrations correlated in groundwater entering the estuary, septic tank effluent appeared responsible for the supersaturated concentrations of N(2)O in surface waters. A hyperbolic function fit nitrate and N(2)O concentrations in the water column of the estuary with a maximum supersaturation of approximately 60 nM. From surface water supersaturation we predicted a release of 480 nmol N(2)O m(-2) h(-1) to the atmosphere in the summer. Property plots of salinity vs. bottom-water N(2)O suggested a benthic sink of N(2)O. Consistent with this trend, sediments consumed rather than released N(2)O in most flux measurements. Nutrient loading did not directly alter benthic N(2)O flux, potentially because stratification limited exposure of sediments to nitrate-rich surface waters, but macroalgal cover increased benthic N(2)O consumption. Sediment N(2)O consumption averaged 111 nmol N(2)O m(-2) h(-1) and correlated with oxygen uptake. Losses from the system to the atmosphere and sediments exceeded inputs of N(2)O contaminated groundwater, which suggests missing N(2)O sources.

Assessment of a delta15N isotopic method to indicate anthropogenic eutrophication in aquatic ecosystems.

Increased anthropogenic delivery of nutrients to water bodies, both freshwater and estuarine, has caused detrimental changes in habitat, food web structure, and nutrient cycling. Nitrogen-stable isotopes may be suitable indicators of such increased nutrient delivery. In this study, we looked at the differences in response of macrophyte delta15N values to anthropogenic N across different taxonomic groups and geographic regions to test a stable isotopic method for detecting anthropogenic impacts. Macrophyte delta15N values increased with wastewater input and water-column dissolved inorganic nitrogen (DIN) concentration. When macrophytes were divided into macroalgae and plants, they responded similarly to increases in wastewater N, although macroalgae was a more reliable indicator of both wastewater inputs and water-column DIN concentrations. Smooth cordgrass (Spartina alterniflora Loisel.) Delta15N increased uniformly with wastewater inputs across a geographic range. We used the relationship derived between S. alterniflora and relative wastewater load to predict wastewater loads in locations lacking quantitative land use data. The predictions matched well with known qualitative information, proving the use of a stable isotopic method for predicting wastewater input.