Developing performance measures of mangrove wetlands using simulation models of hydrology, nutrient biogeochemistry, and community dynamics

The goal of mangrove restoration projects should be to improve community structure and ecosystem function of degraded coastal landscapes. This requires the ability to forecast how mangrove structure and function will respond to prescribed changes in site conditions including hydrology, topography, and geophysical energies. There are global, regional, and local factors that can explain gradients of regulators (e.g., salinity, sulfides), resources (nutrients, light, water), and hydroperiod (frequency, duration of flooding) that collectively account for stressors that result in diverse patterns of mangrove properties across a variety of environmental settings. Simulation models of hydrology, nutrient biogeochemistry, and vegetation dynamics have been developed to forecast patterns in mangroves in the Florida Coastal Everglades. These models provide insight to mangrove response to specific restoration alternatives, testing causal mechanisms of system degradation. We propose that these models can also assist in selecting performance measures for monitoring programs that evaluate project effectiveness. This selection process in turn improves model development and calibration for forecasting mangrove response to restoration alternatives. Hydrologic performance measures include soil regulators, particularly soil salinity, surface topography of mangrove landscape, and hydroperiod, including both the frequency and duration of flooding. Estuarine performance measures should include salinity of the bay, tidal amplitude, and conditions of fresh water discharge (included in the salinity value). The most important performance measures from the mangrove biogeochemistry model should include soil resources (bulk density, total nitrogen, and phosphorus) and soil accretion. Mangrove ecology performance measures should include forest dimension analysis (transects and/or plots), sapling recruitment, leaf area index, and faunal relationships. Estuarine ecology performance measures should include the habitat function of mangroves, which can be evaluated with growth rate of key species, habitat suitability analysis, isotope abundance of indicator species, and bird census. The list of performance measures can be modified according to the model output that is used to define the scientific goals during the restoration planning process that reflect specific goals of the project.

Dynamics of ecosystem metabolism and flocculent detritus transport in estuarine Taylor River

Estuaries and estuarine wetlands are ecologically and societally important systems, exhibiting high rates of primary production that fuel offshore secondary production. Hydrological processes play a central role in shaping estuarine ecosystem structure and function by controlling nutrient loading and the relative contributions of marine and terrestrial influences on the estuary. The Comprehensive Everglades Restoration Plan includes plans to restore freshwater delivery to Taylor Slough, a shallow drainage basin in the southern Everglades, ultimately resulting in increased freshwater flow to the downstream Taylor River estuary. The existing seasonal and inter-annual variability of water flow and source in Taylor River affords the opportunity to investigate relationships between ecosystem function and hydrologic forcing. Estimates of aquatic ecosystem metabolism, derived from free-water, diel changes in dissolved oxygen, were combined with assessments of wetland flocculent detritus quality and transport within the context of seasonal changes in Everglades hydrology. Variation in ecosystem gross primary production and respiration were linked to seasonal changes in estuarine water quality using multiple autoregression models. Furthermore, Taylor River was observed to be net heterotrophic, indicating that an allochthonous source of carbon maintained ecosystem respiration in excess of autochthonous primary production. Wetland-derived detritus appears to be an important vector of energy and nutrients across the Everglades landscape; and in Taylor River, is seasonally flushed into ponded segments of the river where it is then respired. Lastly, seasonal water delivery appears to govern feedbacks regulating water column phosphorus availability in the Taylor River estuary.

Organic matter dynamics in the Florida coastal Everglades: A molecular marker and isotopic approach

Everglades National Park (ENP) is about to undergo the world's largest wetland restoration with the aim of improving the quality, timing and distribution of water flow. The changes in water flow are hypothesized to alter the nutrient fluxes and organic matter (OM) dynamics within ENP, especially in the estuarine areas. This study used a multi-proxy approach of molecular markers and stable δ 13C isotope measurements, to determine the present day OM dynamics in ENP. ^ OM dynamics in wetland soils/sediments have proved to be difficult to understand using traditional geochemical approaches. These are often inadequate to describe the multitude of OM sources (e.g. higher land plant, emergent vegetation, submerged vegetation) to the soils/sediments and the complex diagenetic processes that can alter the OM characteristics. A multi-proxy approach, however, that incorporates both molecular level and bulk parameter information is ideal to comprehend complex OM dynamics in aquatic environments. Therefore, biomass-specific molecular markers or proxies can be useful in tracing the sources and processing of OM. This approach was used to examine the OM dynamics in the two major drainage basins, Shark River Slough and Taylor River Slough, of ENP. Freshwater to marine transects were sampled in both systems for soils/sediments and suspended particulate organic matter (SPOM) to be characterized through bulk OM analyses, lipid biomarker determinations (e.g. sterols, fatty acids, hydrocarbons and triterpenoids) and compound-specific stable carbon isotope (δ 13C) determinations. ^ One key accomplishment of the research was the assessment of a molecular marker proxy (Paq) to distinguish between emergent/higher plant vegetation from submerged vegetation within ENP. This proxy proved to be quite useful at tracing OM inputs to the soils/sediments of ENP. A second key accomplishment was the development of a 3-way model using vegetation specific molecular markers. This novel, descriptive model was successfully applied to the estuarine areas of Taylor and Shark River sloughs, providing clear evidence of mixing of freshwater, estuarine and marine derived OM in these areas. In addition, diagenetic transformations of OM in these estuaries were found to be quite different between Taylor and Shark Rivers, and are likely a result of OM quality and hydrological differences. ^

Quantifying nitrogen cycling rates in freshwater marshes of the southern Everglades using nitrogen-15 tracer techniques

This dissertation research project addressed the question of how hydrologic restoration of the Everglades is impacting the nutrient dynamics of marsh ecosystems in the southern Everglades. These effects were analyzed by quantifying nitrogen (N) cycle dynamics in the region. I utilized stable isotope tracer techniques to investigate nitrogen uptake and cycling between the major ecosystem components of the freshwater marsh system. I recorded the natural isotopic signatures (δ15N and δ 13C) for major ecosystem components from the three major watersheds of the Everglades: Shark River Slough, Taylor Slough, and C-111 basin. Analysis of δ15 N and δ13C natural abundance data were used to demonstrate the spatial extent to which nitrogen from anthropogenic or naturally enriched sources is entering the marshes of the Everglades. In addition, I measured the fluxes on N between various ecosystem components at both near-canal and estuarine ecotone locations. Lastly, I investigated the effect of three phosphorus load treatments (0.00 mg P m-2, 6.66 mg P m-2, and 66.6 mg P m-2) on the rate and magnitude of ecosystem N-uptake and N-cycling. The δ15N and δ13C natural abundance data supported the hypothesis that ecosystem components from near-canal sites have heavier, more enriched δ 15N isotopic signatures than downstream sites. The natural abundance data also showed that the marshes of the southern Everglades are acting as a sink for isotopically heavier, canal-borne dissolved inorganic nitrogen (DIN) and a source for "new" marsh derived dissolved organic nitrogen (DON). In addition, the 15N mesocosm data showed the rapid assimilation of the 15N tracer by the periphyton component and the delayed N uptake by soil and macrophyte components in the southern Everglades.

Relative significance of pelagic, benthic and allochthonous organic matter to microbial carbon cycling in a phosphorus-depleted subtropical estuary (Florida Bay)

Heterotrophic bacteria are important decomposers and transformers of primary production and provide an important link between detritus and the aquatic food web. In seagrass ecosystems, much of seagrass primary production is unavailable through direct grazing and must undergo microbial reworking before seagrass production can enter the aquatic food web. The goal of my dissertation research is to understand better the role heterotrophic bacteria play in carbon cycling in seagrass estuaries. My dissertation research focuses on Florida Bay, a seagrass estuary that has experienced recent changes in carbon source availability, which may have altered ecosystem function. My dissertation research investigates the importance of seagrass, algal and/or cyanobacterial, and allochthonous-derived organic matter to heterotrophic bacteria in Florida Bay and helps establish the carbon base of the estuarine food web. ^ A three tiered approach to the study of heterotrophic bacterial carbon cycling and trophic influences in Florida Bay was used: (1) Spatiotemporal observations of environmental parameters (hydrology, nutrients, extracellular enzymes, and microbial abundance, biomass, and production); (2) Microbial grazing experiments under different levels of top-down and bottom-up influence; and (3) Bulk and compound-specific (bacteria-biomarker fatty acid analysis) stable carbon isotope analysis. ^ In Florida Bay, spatiotemporal patterns in microbial extracellular enzyme (also called ectoenzyme) activities indicate that microorganisms hydrolyzed selectively fractions of the estuarine organic matter pool. The microbial community hydrolyzed organic acids, peptides, and phosphate esters and did not use storage and structural carbohydrates. Organic matter use by heterotrophic bacterioplankton in Florida Bay was co-regulated by bottom-up (resource availability) and top-down (grazer mediated) processes. A bacterial carbon budget based on bacterial, epiphytic, and seagrass production indicates that heterotrophic bacterial carbon cycles are supported primarily through epiphytic production with mixing from seagrass production. Stable carbon isotope analysis of bacteria biomarkers and carbon sources in Florida Bay corroborate the results of the bacterial carbon budget. These results support previous studies of aquatic consumers in Florida Bay, indicating that epiphytic/benthic algal and/or cyanobacterial production with mixing from seagrass-derived organic matter is the carbon base of the seagrass estuarine food web. ^

Hydrological dynamics between a coastal aquifer and the adjacent estuarine system, Biscayne Bay, South Florida

Geochemical and geophysical approaches have been used to investigate the freshwater and saltwater dynamics in the coastal Biscayne Aquifer and Biscayne Bay. Stable isotopes of oxygen and hydrogen, and concentrations of Sr2+ and Ca2+ were combined in two geochemical mixing models to provide estimates of the various freshwater inputs (precipitation, canal water, and groundwater) to Biscayne Bay and the coastal canal system in South Florida. Shallow geophysical electromagnetic and direct current resistivity surveys were used to image the geometry and stratification of the saltwater mixing zone in the near coastal (less than 1km inland) Biscayne Aquifer. The combined stable isotope and trace metal models suggest a ratio of canal input-precipitation-groundwater of 38%–52%–10% in the wet season and 37%–58%–5% in the dry season with an error of 25%, where most (20%) of the error was attributed to the isotope regression model, while the remaining 5% error was attributed to the Sr2+/Ca2+ mixing model. These models suggest rainfall is the dominate source of freshwater to Biscayne Bay. For a bay-wide water budget that includes saltwater and freshwater mixing, fresh groundwater accounts for less than 2% of the total input. A similar Sr 2+/Ca2+ tracer model indicates precipitation is the dominate source in 9 out of 10 canals that discharge into Biscayne Bay. The two-component mixing model converged for 100% of the freshwater canal samples in this study with 63% of the water contributed to the canals coming from precipitation and 37% from groundwater inputs ±4%. There was a seasonal shift from 63% precipitation input in the dry season to 55% precipitation input in the wet season. The three end-member mixing model converged for only 60% of the saline canal samples possibly due to non-conservative behavior of Sr2+ and Ca2+ in saline groundwater discharging into the canal system. Electromagnetic and Direct Current resistivity surveys were successful at locating and estimating the geometry and depth of the freshwater/saltwater interface in the Biscayne Aquifer at two near coastal sites. A saltwater interface that deepened as the survey moved inland was detected with a maximum interpreted depth to the interface of 15 meters, approximately 0.33 km inland from the shoreline. ^

Physiological performance measures and tolerance limits of estuarine indicator species in south Florida

Current water management practices in South Florida have negatively impacted many species inhabiting Florida Bay. Variable and high salinity has been identified as a key stressor in these estuaries. The Comprehensive Everglades Restoration Plan (CERP) includes water redistribution projects that will restore natural freshwater flows to northeastern Florida Bay. My studies focused on the following central theme and hypotheses: Biological performance measures (i.e., growth, reproduction, survival), behavior (i.e., habitat preference and locomotor behavior) and diversity of estuarine fish will be controlled by changes in salinity and water quality that will occur as a result of the restoration of freshwater flow to the bay. A series of acute and subchronic physiological toxicity studies were conducted to determine the effects of salinity changes on the life stages (embryo/larval, juvenile, adult) and fecundity of four native estuarine fish (Cyprinodon variegatus, Floridichthys carpio, Poecilia latipinna, and Gambusia holbrooki). Fish were exposed to a range of salinity concentrations (freshwater to hypersaline) based on salinity profiles in the study areas. Growth (length, weight) and survival were measured. Salinity trials included both rapid and gradual change events. Results show negative effects of acute, abrupt salinity changes on fish survival, development and reproductive success as a result of salinity stress. Other studies targeted reproduction and critical embryo-larval/neonate development as key areas for detecting long-term population effects of salinity change in Florida Bay. Adults of C. variegates and P. latipinna were also examined for behavioral responses to pulsed salinity changes. These responses include changes in swimming performance, locomotor behavior and zone preference. Finally, an ecological risk assessment was conducted for adverse salinity conditions in northeastern Florida Bay. Using the U.S. EPA's framework, the risk to estuarine fish species diversity was assessed against regional salinity profiles from a 17-year database. Based on the risk assessment, target salinity profiles for these areas are recommended for managers.^

Local biomass control on the composition and reactivity of particulate organic matter in aquatic environments

Freshwater ecosystems have been recognized as important components of the global carbon cycle, and the flux of organic matter (OM) from freshwater to marine environments can significantly affect estuarine and coastal productivity. The focus of this study was the assessment of carbon dynamics in two aquatic environments, namely the Florida Everglades and small prairie streams in Kansas, with the aim of characterizing the biogeochemistry of OM. In the Everglades, particulate OM (POM) is mostly found as a layer of flocculent material (floc). While floc is believed to be the main energy source driving trophic dynamics in this oligotrophic wetland, not much is known about its biogeochemistry. The objective of this study was to determine the origin/sources of OM in floc using biomarkers and pigment-based chemotaxonomy to assess specific biomass contributions to this material, on a spatial (freshwater marshes vs. mangrove fringe) and seasonal (wet vs. dry) scales. It was found that floc OM is derived from the local vegetation (mainly algal components and macrophyte litter) and its composition is controlled by seasonal drivers of hydrology and local biomass productivity. Photo-reactivity experiments showed that light exposure on floc resulted in photo-dissolution of POC with the generation of significant amounts of both dissolved OM (DOM) and nutrients (N & P), potentially influencing nutrient dynamics in this ecosystem. The bio-reactivity experiments determined as the amount and rate of CO2 evolution during incubation were found to vary on seasonal and spatial scales and were highly influenced by phosphorus limitation. Not much is known on OM dynamics in small headwater streams. The objective of this study was to determine carbon dynamics in sediments from intermittent prairie streams, characterized by different vegetation cover for their watershed (C4 grasses) vs. riparian zone (C3 plants). In this study sedimentary OM was characterized using a biomarker and compound specific carbon stable isotope approach. It was found that the biomarker composition of these sediments is dominated by higher plant inputs from the riparian zone, although inputs from adjacent prairie grasses were also apparent. Conflicting to some extent with the River Continuum Concept, sediments of the upper reaches contained more degraded OM, while the lower reaches were enriched in fresh material deriving from higher plants and plankton sources as a result of hydrological regimes and particle sorting.

Spatial and temporal patterns of aboveground net primary productivity (ANPP) along two freshwater-estuarine transects in the Florida Coastal Everglades

We present here a 4-year dataset (2001–2004) on the spatial and temporal patterns of aboveground net primary production (ANPP) by dominant primary producers (sawgrass, periphyton, mangroves, and seagrasses) along two transects in the oligotrophic Florida Everglades coastal landscape. The 17 sites of the Florida Coastal Everglades Long Term Ecological Research (FCE LTER) program are located along fresh-estuarine gradients in Shark River Slough (SRS) and Taylor River/C-111/Florida Bay (TS/Ph) basins that drain the western and southern Everglades, respectively. Within the SRS basin, sawgrass and periphyton ANPP did not differ significantly among sites but mangrove ANPP was highest at the site nearest the Gulf of Mexico. In the southern Everglades transect, there was a productivity peak in sawgrass and periphyton at the upper estuarine ecotone within Taylor River but no trends were observed in the C-111 Basin for either primary producer. Over the 4 years, average sawgrass ANPP in both basins ranged from 255 to 606 g m−2 year−1. Average periphyton productivity at SRS and TS/Ph was 17–68 g C m−2 year−1 and 342–10371 g C m−2 year−1, respectively. Mangrove productivity ranged from 340 g m−2 year−1 at Taylor River to 2208 g m−2 year−1 at the lower estuarine Shark River site. Average Thalassia testudinum productivity ranged from 91 to 396 g m−2 year−1 and was 4-fold greater at the site nearest the Gulf of Mexico than in eastern Florida Bay. There were no differences in periphyton productivity at Florida Bay. Interannual comparisons revealed no significant differences within each primary producer at either SRS or TS/Ph with the exception of sawgrass at SRS and the C−111 Basin. Future research will address difficulties in assessing and comparing ANPP of different primary producers along gradients as well as the significance of belowground production to the total productivity of this ecosystem.

Benthic Macrophyte Distribution and Abundance in Estuarine Mangrove Lakes and Estuaries: Relationships to Environmental Variables

Annual mean salinity, light availability, and sediment depth to bedrock structured the submerged aquatic vegetation (SAV) communities in subtropical mangrove-lined estuaries. Three distinct SAV communities (i.e., Chara group, Halodule group, and Low SAV coverage group) were identified along the Everglades–Florida Bay ecotone and related to water quality using a discriminant function model that predicted the type of plant community at a given site from salinity, light availability, and sediment depth to bedrock. Mean salinity alone was able to correctly classify 78% of the sites and reliably separated the Chara group from the Halodule group. The addition of light availability and sediment depth to bedrock increased model accuracy to 90% and further distinguished the Chara group from the Halodule group. Light availability was uniquely valuable in separating the Chara group from the Low SAV coverage group. Regression analyses identified significant relationships between phosphorus concentration, phytoplankton abundance, and light availability and suggest that a decline in water transparency, associated with increasing salinity, may have also contributed to the historical decline of Chara communities in the region. This investigation applies relationships between environmental variables and SAV distribution and provides a case study into the application of these general principals to ecosystem management.

Spatial patterns of fish communities along two estuarine gradients in southern Florida

In tropical and subtropical estuaries, gradients of primary productivity and salinity are generally invoked to explain patterns in community structure and standing crops of fishes. We documented spatial and temporal patterns in fish community structure and standing crops along salinity and nutrient gradients in two subtropical drainages of Everglades National Park, USA. The Shark River drains into the Gulf of Mexico and experiences diurnal tides carrying relatively nutrient enriched waters, while Taylor River is more hydrologically isolated by the oligohaline Florida Bay and experiences no discernable lunar tides. We hypothesized that the more nutrient enriched system would support higher standing crops of fishes in its mangrove zone. We collected 50 species of fish from January 2000 to April 2004 at six sampling sites spanning fresh to brackish salinities in both the Shark and Taylor River drainages. Contrary to expectations, we observed lower standing crops and density of fishes in the more nutrient rich tidal mangrove forest of the Shark River than in the less nutrient rich mangrove habitats bordering the Taylor River. Tidal mangrove habitats in the Shark River were dominated by salt-tolerant fish and displayed lower species richness than mangrove communities in the Taylor River, which included more freshwater taxa and yielded relatively higher richness. These differences were maintained even after controlling for salinity at the time of sampling. Small-scale topographic relief differs between these two systems, possibly created by tidal action in the Shark River. We propose that this difference in topography limits movement of fishes from upstream marshes into the fringing mangrove forest in the Shark River system, but not the Taylor River system. Understanding the influence of habitat structure, including connectivity, on aquatic communities is important to anticipate effects of construction and operational alternatives associated with restoration of the Everglades ecosystem.

Biogeochemistry of Nitrogen Across the Everglades Landscape

Compared to phosphorus (P), nitrogen (N) has received little attention across the Everglades landscape. Despite this lack of attention, N plays important roles in many Everglades systems, including being a significant pollutant in Florida Bay and the Gulf of Mexico, the limiting nutrient in highly P-impacted areas, and an important substrate for microbial metabolism. Storage and transport of N throughout the Everglades is dominated by organic forms, including peat soils and dissolved organic N in the water column. In general, N sources are highest in the northern areas; however, atmospheric deposition and active N2 fixation by the periphyton components are a significant N source throughout most systems. Many of the processes involved in the wetland N cycle remain unmeasured for most of the Everglades systems. In particular, the lack of in situ rates for N2 fixation and denitrification prevent the construction of system-level budgets, especially for the Southern mangrove systems where N export into Florida Bay is critical. There is also the potential for several novel N processes (e.g., Anammox) with an as yet undetermined importance for nitrogen cycling and function of the Everglades ecosystem. Phosphorus loading alters the N cycle by stimulating organic N mineralization with resulting flux of ammonium and DON, and at elevated P concentrations, by increasing rates of N2 fixation and N assimilation. Restoration of hydrology has a potential for significantly impacting N cycling in the Everglades both in terms of affecting N transport, but also by altering aerobic-anaerobic transitions at the soil-water interface or in areas with seasonal drawdowns (e.g., marl prairies). Based on the authors’ understanding of N processes, much more research is necessary to adequately predict potential impacts from hydrologic restoration, as well as the function of Everglades systems as sinks, sources, and transformers of N in the South Florida landscape.

Chemical characteristics of dissolved organic matter in an oligotrophic subtropical wetland/estuarine ecosystem

Fluorescence properties of whole water samples and molecular characteristics of ultrafiltrated dissolved organic matter (UDOM > 1,000 D) such as lignin phenol and neutral sugar compositions and 13C nuclear magnetic resonance (NMR) spectra were determined along a freshwater to marine gradient in Everglades National Park. Furthermore, UDOM samples were categorized by hierarchical cluster analysis based on their pyrolysis gas chromatography/mass spectrometry products. Fluorescence properties suggest that autochthonous DOM leached/exuded from biomass is quantitatively important in this system. 13C NMR spectra showed that UDOM from the oligotrophic Taylor Slough (TS) and Florida Bay (FB) ecosystems has low aromatic C (13% ± 3% for TS; 2% ± 2% for FB) and very high O-alkyl C (54% ± 4% for TS; 75% ± 4% for FB) concentrations. High O-alkyl C concentrations in FB suggest seagrass/phytoplankton communities as dominant sources of UDOM. The amount of neutral sugars was not appreciably different between the TS and FB sites (115 ± 12 mg C g C-1 UDOM) but their concentrations suggest a low level of diagenesis and high production rates of this material in this oligotrophic environment. Total yield of lignin phenols (vanillyl + syringyl phenols) in TS was low (0.20–0.39 mg 100 mg C-1 UDOM) compared with other riverine environments and even lower in FB (0.04–0.07 mg 100 mg C-1 UDOM) and could be a result of photodegradation and/or dilution by other utochthonous DOM. The high O-alkyl and low aromatic nature of this UDOM suggests significant biogenic inputs (as compared with soils) and limited bioavailability in this ecosystem.

Chemical characterization of dissolved organic nitrogen in an oligotrophic subtropical coastal ecosystem

Water samples were collected from rivers and estuarine environments within the Florida Coastal Everglades (FCE) ecosystem, USA, and ultrafiltered dissolved organic matter (UDOM; 1 kDa) was isolated for characterization of its source, bioavailability and diagenetic state. A combination of techniques, including 15N cross-polarization magic angle spinning nuclear magnetic resonance (15N CPMAS NMR) and X-ray photoelectron spectroscopy (XPS), were used to analyze the N components of UDOM. The concentrations and compositions of total hydrolysable amino acids (HAAs) were analyzed to estimate UDOM bioavailability and diagenetic state. Optical properties (UV–visible and fluorescence) and the stable isotope ratios of C and N were measured to assess the source and dynamics of UDOM. Spectroscopic analyses consistently showed that the major N species of UDOM are in amide form, but significant contributions of aromatic-N were also observed. XPS showed a very high pyridinic-N concentration in the FCE–UDOM (21.7 ± 2.7%) compared with those in other environments. The sources of this aromatic-N are unclear, but could include soot and charred materials from wild fires. Relatively high total HAA concentrations (4 ± 2% UDOC or 27 ± 4% UDON) are indicative of bioavailable components, and HAA compositions suggest FCE–UDOM has not undergone extensive diagenetic processing. These observations can be attributed to the low microbial activity and a continuous supply of fresh UDOM in this oligotrophic ecosystem. Marsh plants appear to be the dominant source of UDOM in freshwater regions of the FCE, whereas seagrasses and algae are the dominant sources of UDOM in Florida Bay. This study demonstrates the utility of a multi-technique and multi-proxy approach to advance our understanding of DON biogeochemistry.

Composition of a protein-like fluorophore of dissolved organic matter in coastal wetland and estuarine ecosystems

This study demonstrates the compositional heterogeneity of a protein-like fluorescence emission signal (T-peak; excitation/emission maximum at 280/325 nm) of dissolved organic matter (DOM) samples collected from subtropical river and estuarine environments. Natural water samples were collected from the Florida Coastal Everglades ecosystem. The samples were ultrafiltered and excitation–emission fluorescence matrices were obtained. The T-peak intensity correlated positively with N concentration of the ultrafiltered DOM solution (UDON), although, the low correlation coefficient (r2=0.140, p<0.05) suggested the coexistence of proteins with other classes of compounds in the T-peak. As such, the T-peak was unbundled on size exclusion chromatography. The elution curves showed that the T-peak was composed of two compounds with distinct molecular weights (MW) with nominal MWs of about >5×104 (T1) and ∼7.6×103 (T2) and with varying relative abundance among samples. The T1-peak intensity correlated strongly with [UDON] (r2=0.516, p<0.001), while T2-peak did not, which suggested that the T-peak is composed of a mixture of compounds with different chemical structures and ecological roles, namely proteinaceous materials and presumably phenolic moieties in humic-like substances. Natural source of the latter may include polyphenols leached from senescent plant materials, which are important precursors of humic substances. This idea is supported by the fact that polyphenols, such as gallic acid, an important constituent of hydrolysable tannins, and condensed tannins extracted from red mangrove (Rhizophora mangle) leaves exhibited the fluorescence peak in the close vicinity of the T-peak (260/346 and 275/313 nm, respectively). Based on this study the application of the T-peak as a proxy for [DON] in natural waters may have limitations in coastal zones with significant terrestrial DOM input.