Elizabeth River Research Articles

Enrichment, Isolation, and Phylogenetic Identification of Polycyclic Aromatic Hydrocarbon-Degrading Bacteria from Elizabeth River Sediments

The diversity of indigenous bacteria in sediments from several sites in the Elizabeth River (Virginia) able to degrade multiple polycyclic aromatic hydrocarbons (PAHs) was investigated by the use of classical selective enrichment and molecular analyses. Enrichment cultures containing naphthalene, phenanthrene, fluoranthene, or pyrene as a sole carbon and energy source were monitored by denaturing gradient gel electrophoresis (DGGE) to detect changes in the bacterial-community profile during enrichment and to determine whether the representative strains present were successfully cultured. The DGGE profiles of the final enrichments grown solely on naphthalene and pyrene showed no clear relationship with the site from which the inoculum was obtained. The enrichments grown solely on pyrene for two sample sites had >80% similarity, which suggests that common pyrene-degrading strains may be present in these sediments. The final enrichments grown on fluoranthene and phenanthrene remained diverse by site, suggesting that these strains may be influenced by environmental conditions. One hundred and one isolates were obtained, comprising representatives of the actinomycetes and alpha-, beta-, and gammaproteobacteria, including seven novel isolates with 16S rRNA gene sequences less than 98% similar to known strains. The ability to degrade multiple PAHs was demonstrated by mineralization of 14C-labeled substrate and growth in pure culture. This supports our hypothesis that a high diversity of bacterial strains with the ability to degrade multiple PAHs can be confirmed by the combined use of classical selective enrichment and molecular analyses. This large collection of diverse PAH-degrading strains provides a valuable resource for studies on mechanisms of PAH degradation and bioremediation.

Working on the Elizabeth River

A fragile and contaminated ecosystem, intense industrial activity, and human inhabitation struggle to coexist along the Elizabeth River. In three intertwined projects, we have developed translatable urban and architectural strategies interdependent with ecological regeneration, while demonstrating the didactic value of design in public environmental education. This design research model operates at multiple scales, reaches out to diverse communities, plans for varying, overlapping periods of time, and makes positive change in the world.

Concentrations of Metals in Blood and Feathers of Nestling Ospreys (Pandion haliaetus) in Chesapeake and Delaware Bays

In 2000, 2001, and 2002, blood and feather samples were collected from 40-45-day-old nestling ospreys (Pandion haliaetus) from Chesapeake Bay and Delaware Bay and River. Concentrations of 18 metals, metalloids, and other elements were determined in these samples by inductively coupled plasma-mass spectroscopy, and Hg concentrations were measured by cold vapor atomic absorption spectroscopy. When compared to concurrent reference areas (South, West, and Rhode Rivers), mean As and Hg concentrations in blood were greater (p < 0.05) in two of three Chesapeake Bay regions of concern (Baltimore Harbor [As: 1.18 vs. 0.548 microg/g dw], Anacostia River [Hg: 0.305 vs. 0.178 microg/g dw], and Elizabeth River [As: 0.876 vs. 0.663 microg/g dw; Hg: 0.260 vs. 0.180 microg/g dw]). Lead was detected more frequently in blood of nestlings from the highly industrialized Elizabeth River compared to the rural reference area. When compared to the concurrent reference area, mean Al, Ba, Hg, Mn, and Pb concentrations in feathers were substantially greater (p < 0.05) in one or more Chesapeake regions of concern (Anacostia River [Al: 206 vs. 62.1 microg/g dw; Ba: 3.31 vs. 0.823 microg/g dw; Mn: 65.4 vs. 22.9 microg/g dw] and Elizabeth River [Al: 165 vs. 63.5 microg/g dw; Hg: 1.24 vs. 0.599 microg/g dw; Pb 1.47 vs. 0.543 microg/g dw]). When compared to the coastal Inland Bays reference area, feathers of nestlings from northern Delaware Bay and River had greater concentrations (p < 0.05) of Ba (1.90 vs. 0.660 microg/g dw), Fe (258 vs. 109 microg/g dw), Mn (18.5 vs. 4.66 microg/g dw), Mo (0.130 vs. 0.040 microg/g dw), Pb (1.96 vs. 0.624 microg/g dw), and V (0.671 vs. 0.325 microg/g dw), presumably due to extensive metal-working and petroleum refinery activities. Concentrations of Hg in nestling feathers from Delaware were frequently greater than in the Chesapeake. The present findings and those of related reproductive studies suggest that concentrations of several heavy metals (e.g., Cd, Hg, Pb) in nestling blood and feathers from Chesapeake and Delaware Bays were below toxicity thresholds and do not seem to be affecting chick survival during the nestling period.

Bioindicators of Immune Function in Creosote-adapted Estuarine Killifish, Fundulus heteroclitus

Several populations of the estuarine killifish, Fundulus heteroclitus, also known as the mummichog, exhibit characteristics of adaptation to priority pollutants. One such population of mummichog inhabits the Elizabeth River in Virginia at the Atlantic Wood site (AW), a U.S. Environmental Protection Agency (EPA) Superfund site heavily contaminated with creosote containing a mixture of polyaromatic hydrocarbons (PAHs). Although PAHs are known to be immunotoxic in experimental animals, resident AW mummichogs seem to thrive. Mummichogs from the AW site and a reference site were subsequently examined over a 2-yr period for total immunoglobin (IgM), as well as circulating antibody levels against 5 ubiquitous marine bacteria. Expression profiles of circulating and lymphoid lysozyme and lymphoid cyclooxygenase-2 (COX-2) were also examined. Compared to relatively high total IgM and specific antibody responses in reference fish, AW mummichogs had lower circulating IgM and lower specific antibody levels against all bacteria examined, however they had higher levels of circulating lysozyme. Lymphoid cells in the AW mummichogs also expressed higher levels of lysozyme, as well as COX-2, which may indicate a state of macrophage activation. Elevated COX-2 levels may be associated with enhanced metabolism of PAHs through cooxidation-peroxidase pathways. Additional studies attempted to immunize AW mummichogs reared in uncontaminated water to compare their antibody responses to that of reference fish. AW mummichogs did not survive 40 d post culture, while reference fish thrived. Our findings suggest that the chemical environment at the AW site may be vicariously enhancing components of innate immunity, probably through oxidative stress pathways, in resident mummichogs, while actively suppressing humoral immune responses.

Depth-related patterns in benthic community condition along an estuarine gradient in Chesapeake Bay, USA

Abstract We tested whether macrobenthic community condition varies significantly with water depth in a variety of regions of Chesapeake Bay, USA. Benthic community condition was characterized using the Benthic Index of Biotic Integrity (B-IBI) previously developed for the Bay. We applied two water depth thresholds intended to emphasize the ecological importance and/or anthropogenic impacts upon shallow-water regions. The first threshold of 2 m emphasizes restoring and supporting submerged aquatic vegetation while the second threshold of 4 m emphasizes the zone of maximum anthropogenic impact upon natural ecosystem functions. An a priori expectation is that benthic community condition may worsen with increasing depth, specifically in regions (1) where water column stratification at depth results in prolonged low dissolved oxygen levels or (2) where net deposition at depth results in higher levels of hydrophobic, sediment-bound contaminants. Samples collected from a major tributary of Chesapeake Bay, the York River estuary, spanned the entire salinity range from tidal freshwater to polyhaline. We also tested the shallow-water depth thresholds using data from the Virginia Mainstem of Chesapeake Bay and the Southern Branch of the Elizabeth River. These two polyhaline regions are characterized as having the best and worst benthic community condition in Chesapeake Bay. At the scale of the entire tidal York River system, there were no significant differences in benthic community condition with water depth. However, two salinity regions, low mesohaline and polyhaline, had significant depth effects with the shallowest water depth zone significantly different from the other two depth regions. For the low mesohaline region benthic community condition was worse at the shallowest depth and for the polyhaline region the shallowest depth was better comparing the three depth regions. No depth-related differences in the B-IBI were found for the two additional Chesapeake Bay strata, the Virginia Mainstem characterized with the lowest levels of benthic community degradation and for the Southern branch of the Elizabeth River, characterized by the highest levels of benthic community degradation. We conclude that the ecological state of Chesapeake Bay subtidal benthic communities is adequately characterized by randomly sampling all depths without further stratification into shallow and deeper regions.

The effect of maternal exposure to contaminated sediment on the growth and condition of larval Fundulus heteroclitus

We employed a factorial laboratory experiment to determine the single and combined effect of maternal and larval exposure to contaminated sediment from Elizabeth River, Virginia, a site contaminated with high concentrations of multiple pollutants. Females were exposed to either reference or contaminated sediment and the larvae from both groups of mothers were in turn transferred to either reference or contaminated sediment. We found a strong maternal influence on yolk area, length and RNA:DNA ratio at hatch. Further, the maternal exposure significantly influenced growth rate and RNA:DNA ratios of larvae 14 days after hatch and was a more important factor in determining these endpoints than larval exposure. We found that after 14 days larvae were larger and had higher survivorship when the maternal and larval exposures were the same. There also was no statistical difference with respect to growth and condition between larvae that had hatched from exposed mothers and remained in contaminated water and larvae that had hatched from reference mothers and were placed in either reference or contaminated sediment. However, larvae that hatched from exposed mothers and then were switched to reference sediment had significantly lower growth, lower RNA:DNA ratios, and were smaller despite being large at hatch size, indicating that there are fitness trade-offs in exchange for apparent resistance to contaminants which are provided by the mother. Maternal effects add complexity to ecotoxicological research and should be incorporated into studies to predict population level responses more realistically.

Assessment of the historical trace metal contamination of sediments in the Elizabeth River, Virginia

Two sediment cores (Southern Branch, PC-1, and Western Branch, WB-2) were taken from the highly industrialized Elizabeth River, Virginia. The concentrations of trace metals cadmium, cobalt, chromium, copper, nickel, lead and zinc, major elements iron, manganese and aluminum, organic carbon content and the specific surface area of the sediments were determined in each of the cores. Down-core variations in metals varied significantly in each core with maximum contamination events occurring at different times in different portions of the river. In PC-1, maximum metal concentrations were seen after the appearance of 137Cs. In contrast, the highest levels in WB-2 occurred well before the appearance of 137Cs. Although stricter environmental regulations have caused a decrease in metal concentrations since the 1980s, the concentrations in the surface sediments of many trace metals were elevated to levels 2–5 times higher than the levels at the bottom of the cores in both the Southern and Western Branches of the river.

Seasonal survey of copper-complexing ligands and thiol compounds in a heavily utilized, urban estuary: Elizabeth River, Virginia

Concentrations of thiol compounds, copper-complexing ligands, and total dissolved copper were followed over the course of 1 year (October 2002 until September 2003) in the Elizabeth River, Virginia to evaluate seasonality. Copper-complexing ligand concentrations were determined by competitive ligand equilibration-adsorptive cathodic stripping voltammetry (CLE/ACSV). Thiol detection was carried out by high performance liquid chromatography (HPLC) and calibration using a suite of nine thiol compounds (cysteine, glutathione, mercaptoacetic acid, 2-mercaptoethanesulfonic acid, 2-mercaptoethanol, 2-mercaptopropionic acid, 3-mercaptopropionic acid, mercaptosuccinic acid, and monothioglycerol). Total dissolved copper concentrations reached a January low of 13.1 nM to a June high of 24.7 nM and were found to vary seasonally with higher concentrations occurring from June to September. With a low of 26 nM during April to a high of 56 nM in October, copper-complexing ligand (average log K′ CuL of 12.0 ± 0.2) concentrations displayed a similar seasonal pattern to that of total dissolved copper. Free cupric ion concentrations remained below 1.5 pM for a majority of the year except during March, April, and December when values reached pM levels greater than 1.5. Six of the nine thiol compounds surveyed were detected in the Elizabeth River samples and ranged in concentration from below detectable concentrations (< 5 nM) to individual highs ranging from 25.3 to168.5 nM. The thiol compound concentrations displayed a clear seasonality fluctuating at below detection limits during November to February then increasing with increasing surface water temperatures from March to July. CLE/ACSV was used to assess whether or not the suite of thiol compounds detected by HPLC could contribute to the copper-complexing ligand pool. Conditional stability constants for each one of six thiol standards (average log K′ CuL ∼ 12.1 ± 0.5) were found to be statistically equivalent to the naturally occurring copper-complexing ligands (average log K′ CuL ∼ 12.0 ± 0.2). This suggests that these thiol compounds could act as copper-complexing ligands in natural samples and could contribute to the copper-complexing ligand pool detected by CLE/ACSV. This study involving seasonality of copper-complexing ligands and thiols in an industrialized, urban estuary underscored several points that have to be substantiated in future research efforts including copper-complexing ligands sources and the role that thiol compounds as well as other unidentified organic compounds play in the copper-complexing ligand pool.

Assessment of polycyclic aromatic hydrocarbon input to urban wetlands in relation to adjacent land use

The relationship between polycyclic aromatic hydrocarbons (PAHs) in wetland surface sediments and adjacent land use was assessed in the Elizabeth River, VA, an urbanized sub-estuary of the Chesapeake Bay. Significant differences ( p < 0.05) in surface sediment PAH concentration between sites indicated adjacent land use had a substantial influence on PAH concentration in wetland sediments. Wetlands adjacent to parking lots and petroleum industrial sites exhibited the highest PAH concentrations of all wetlands examined. Overall, commercial land uses had the highest PAH concentrations and automotive sources dominated (52–69%) PAH input to wetland surface sediments irrespective of adjacent land use.

Analysis of CpG methylation in the killifish CYP1A promoter

Fundulus heteroclitus (Atlantic killifish or mummichog) inhabiting a creosote-contaminated Superfund site on the Elizabeth River (VA, USA), exhibit a lack of induction of cytochrome P4501A (CYP1A) mRNA, immunodetectable protein, and catalytic activity after exposure to typical inducers. This “refractory CYP1A phenotype” is not explained by alterations in mRNA expression of known CYP1A transcription factors. Furthermore, the refractory phenotype is lost progressively during development in laboratory-reared F 1 generation fish. Thus, while heritable, the refractory CYP1A phenotype does not appear to be genetically based. To test the hypothesis that cytosine methylation at CpG sites in the promoter region of CYP1A underlies the refractory CYP1A phenotype, we employed bisulfite sequencing to compare the methylation status of CpG sites in the CYP1A promoter region of DNA from killifish from the Elizabeth River and a reference site. We examined genomic DNA both from livers of wild-caught adult killifish and from pools of F 1 generation embryos raised in the laboratory. In fish from both the contaminated and the reference site, cytosine methylation was not detectable at any of the 34 CpG sites examined, including 3 that are part of putative xenobiotic response elements.

A PCR-based specific assay reveals a population of bacteria within the Chloroflexi associated with the reductive dehalogenation of polychlorinated biphenyls

Polychlorinated biphenyls (PCBs) accumulate and persist in sediments posing a risk to human health and the environment. Highly chlorinated PCBs are reductively dechlorinated in anaerobic sediments and two bacteria, designated o-17 and DF-1, from a novel phylogenetic group that reductively dechlorinate PCBs have recently been identified. However, there is a paucity of knowledge about the distribution, diversity and ecology of PCB-dechlorinating bacteria due to difficulty in obtaining pure cultures and the lack of detection by universal PCR 16S rRNA gene primer sets in sediments. A specific PCR primer was developed and optimized for detection of o-17/DF-1 and other closely related bacteria in the environment. Using this primer set it was determined that bacteria of this group were enriched in sediment microcosms from Baltimore Harbour concurrent with active dechlorination of 2,2',3,4,4',5'-hexachlorobiphenyl. Additional 16S rRNA gene sequences that had high levels of similarity to described PCB dechlorinators were detected in sediments from the Elizabeth River tributary of Chesapeake Bay, which had confirmed PCB-dechlorinating activities. Phylogenetic comparison of these detected 16S rRNA gene sequences revealed a relatively diverse group of organisms within the dehalogenating Chloroflexi that are distinct from the Dehalococcoides spp. Results from this study indicate that reductive PCB dechlorination activity may be catalysed by a previously undescribed group of micro-organisms that appear to be prevalent in PCB-impacted sites.

Differential display of hepatic mRNA from killifish ( Fundulus heteroclitus) inhabiting a Superfund estuary

Fundulus heteroclitus (Atlantic killifish, mummichog) from a highly contaminated site on the Elizabeth River (VA, USA) are resistant to the toxicity of sediment from the site. However, the mechanistic changes that confer resistance to the toxicity are not yet well understood. We utilized differential display to identify mRNAs that are differentially expressed in hepatic tissue of male and female killifish from the Elizabeth River environment, compared to killifish from a non-contaminated reference site, King's Creek (VA, USA). Seventy-four differentially expressed mRNAs were initially identified (including sex and population-specific differences), and 65 of these were isolated and sequenced. A reverse northern blot array constructed of these cDNAs (plus an additional 15 previously sequenced mRNAs of interest) was used to confirm and quantify expression differences. High interindividual variability was observed in mRNA expression, but multiple differentially expressed mRNAs were identified, including 11 population-specific differences occurring in both sexes, 24 population-specific differences occurring in only one sex, and 22 sex-specific differences. Many of these differentially expressed mRNAs were novel, or not previously hypothesized to play a role in response to contaminant exposure. In addition, the results indicate that the effect of contaminated sediment exposure on the expression of a large proportion of the differentially expressed mRNAs was dependent on the sex of the fish.

Molecular and isotopic identification of PAH sources in a highly industrialized urban estuary

Sources of polycyclic aromatic hydrocarbons (PAHs) in sediments in the Elizabeth River, VA, USA, a highly industrialized urban estuary, were examined using multiple source identification techniques. Large-scale historical creosote releases from former wood-treatment facilities had long been considered the dominant source of PAHs to the estuary. Employing principal component analysis (PCA) along with identification of source-specific isomer ratios, contributions from two former wood-treatment facilities were differentiated. Additionally, a significant coal contribution was determined by also incorporating compound-specific carbon isotope ratio analysis (CSIA). Use of CSIA also successfully isolated and separated the coal signature from a coal gasification signature, which could not be distinguished using only PCA and isomer ratios. The major contributors to sediment PAH contamination in the river were successfully identified, which included a former wood-treatment facility and historical and/or current coal transport and use, a source not considered in previous investigations.

Sources and sinks of dissolved phytochelatin in natural seawater

We modeled dissolved phytochelatin concentrations by studying its exudation from phytoplankton and its removal from seawater. Exudation rates were obtained from experiments in which a marine diatom, Thalassiosira pseudonana, was grown in Cd-containing medium. Dissolved phytochelatin in the culture medium was measured as a function of time, and a first-order model was fit to the data to derive exudation rate constants. A synthetic phytochelatin standard was used to study phytochelatin removal from natural seawater. The addition of Cd or Cu, as well as decreasing temperature, significantly decreased phytochelatin removal rates. An acclimation period followed by rapid removal was observed; subsequent incubations showed no acclimation. Using rate constants derived from these experiments and some assumptions regarding the particulate phytochelatin pool, we used a model to calculate the steady state concentration of dissolved phytochelatin in the field and obtained values that are close to field measurements of dissolved phytochelatin in the Elizabeth River estuary, Norfolk, Virginia, where metal concentrations are high.

Polycyclic aromatic hydrocarbons in a highly industrialized urban estuary: Inventories and trends

The abundance and composition of polycyclic aromatic hydrocarbons (PAHs) in sediments of the main stem and southern branch of the Elizabeth River (VA, USA), a highly industrialized urban estuary in the Chesapeake Bay (USA) watershed, were examined relative to historical and toxic effects levels. Total PAH concentrations in Elizabeth River sediments exceeded those observed in Baltimore Harbor and the Anacostia River, two other regions of concern in the Chesapeake Bay. The sigmaPAH concentrations from samples collected in the vicinity of two former wood-treatment facilities in the Elizabeth River had the highest sigmaPAH when compared to coastal and estuarine systems around the world. Using a linearized diffusion model equation, as much as 69% of the variability in channel sediment sigmaPAH distribution could be ascribed to inputs associated with former wood-treatment facilities along the southern branch of the Elizabeth River. Comparison of PAH levels measured in channel samples to data collected during the early 1980s demonstrated a general trend toward reduction in contaminant concentrations for most regions of the Elizabeth River channel; however, steady-state and increased sediment PAH concentrations in the vicinity of the former wood-treatment facilities were observed. Based on examination of the contaminant levels in Elizabeth River sediments using established sediment-quality criteria, the southern branch of the river remains a clear hazard to benthic and pelagic organisms.

“Cellular” vs. “detrital” POM: a preliminary study using fluorescent stains, flow cytometry, and mass spectrometry

The mass of nonliving particulate organic matter (POM) in marine waters can be significantly greater than that in living cells, yet very little is known about the composition and ultimate fate of this “detrital” POM. In recent marine chemistry studies, flow cytometry has been used to provide further insights into the composition, source, and fate of chlorophyll-fluorescing POM (which includes living algal cells) vs. detrital POM within natural waters. In this study, we extend such morphological characterization of POM beyond the use of chlorophyll autofluorescence and particle size or light scatter. Instead, nucleic acid staining with SYTO-13 and flow cytometry sorting were used to identify and isolate “cellular” and “detrital” subclasses of POM from samples collected in the Lafayette River and the Elizabeth River (subestuaries in the lower Chesapeake Bay). The resulting subclasses, as well as aliquots of unsorted POM sample, were then analyzed by direct temperature-resolved mass spectrometry. We show that “cellular” and “detrital” subclasses differ significantly from each other, and that detrital POM appears chemically distinct from the bulk POM. We then suggest further applications for flow cytometry approaches in aquatic organic biogeochemistry.

Contaminant exposure and reproductive success of ospreys (Pandion haliaetus) nesting in Chesapeake Bay regions of concern.

The Chesapeake Bay osprey population has more than doubled in size since restrictions were placed on the production and use of DDT and other toxic organochlorine contaminants in the 1970s. Ospreys are now nesting in the most highly polluted portions of the Bay. In 2000 and 2001, contaminant exposure and reproduction were monitored in ospreys nesting in regions of concern, including Baltimore Harbor and the Patapsco River, the Anacostia and middle Potomac rivers, and the Elizabeth River, and a presumed reference site consisting of the South, West, and Rhode rivers. A "sample egg" from each study nest was collected for contaminant analysis, and the fate of eggs remaining in each nest (n = 14-16/site) was monitored at 7- to 10-day intervals from egg incubation through fledging of young. Ospreys fledged young in regions of concern (observed success: 0.88-1.53 fledglings/active nest), although productivity was marginal for sustaining local populations in Baltimore Harbor and the Patapsco River and in the Anacostia and middle Potomac rivers. Concentrations of p,p'-DDE and many other organochlorine pesticides or metabolites, total PCBs, some arylhydrocarbon receptor-active PCB congeners and polybrominated diphenyl ether congeners, and perfluorooctanesulfonate were often greater in sample eggs from regions of concern compared to the reference site. Nonetheless, logistic regression analyses did not provide evidence linking marginal productivity to p,p'-DDE, total PCBs, or arylhydrocarbon receptor-active PCB congener exposure in regions of concern. In view of the moderate concentrations of total PCBs in eggs from the reference site, concerns related to new and emerging toxicants, and the absence of ecotoxicological data for terrestrial vertebrates in many Bay tributaries, a more thorough spatial evaluation of contaminant exposure in ospreys throughout the Chesapeake may be warranted.

Interactive regulation of dissolved copper toxicity by an estuarine microbial community

Cultured marine microorganisms under copper stress produce extracellular compounds having a high affinity for copper (copper‐complexing ligands). These ligands are similar in binding strength to those found in natural waters, but few studies have examined the relationship between copper, copper‐complexing ligand concentrations, and natural microbial populations. A series of in situ experiments in the Elizabeth River, Virginia, revealed that an intact estuarine microbial community responded to copper stress by production of extracellular, high‐affinity copper complexing ligands. The rate of ligand production was dependent on copper concentration and resulted in a reduction of the concentration of free cupric ions, Cu2+, by more than three orders of magnitude during a 2‐week period in one experiment. We believe that this interactive response to copper stress represents a feedback system through which microbial communities can potentially buffer dissolved Cu2+ ion concentrations, thereby regulating copper bioavailability and toxicity.

Spatial survey of trace metal contaminants in the sediments of the Elizabeth River, Virginia

The Elizabeth River is a sub-estuary of the James River, the most southern tributary of the Chesapeake Bay. It is a highly industrialized area, and has been designated a “toxic hot spot” due to the heavy loads of contaminant metals and organic compounds in its bed sediments. Fifty surface sediment samples were taken along the channels and shoals of the Mainstem and the Southern Branch portions of the river. The samples were analyzed for trace metals (Cd, Cr, Cu, Pb and Zn), Fe and particle properties (% sand, % silt, % clay and specific surface area) to discern the spatial distribution of contaminant metals in this system. Enrichment factors were calculated to assess the overall level of contamination relative to other surrounding waterways as well as historic contamination levels within the river itself. The highest levels of metals were found to be in close proximity to industrial sources of contamination. The overall level of contamination was 3–10 times higher than baseline levels within the river, and 2–3 times higher than contaminant levels in other area waters.

Submarine groundwater discharge of nutrients and copper to an urban subestuary of Chesapeake Bay (Elizabeth River)

We investigated the role submarine groundwater discharge (SGD) plays in the delivery of nutrients and copper to the Elizabeth River (Virginia) estuary, a major subestuary of lower Chesapeake Bay. Using an approach based on radium isotopes, we concluded that two distinct sources of groundwater were equally impacting the estuary: a surface (marsh) aquifer and deep aquifer source each with a unique 228Ra/226Ra activity ratio. Considering each of these sources, we calculated an SGD flux of 1 x 106 m3 d−1 (±10%), which represented ~6% of the SGD flux for the entire Chesapeake Bay and ~5% of the James River, a major source of freshwater to lower Chesapeake Bay. SGD‐derived dissolved inorganic nitrogen (DIN) and dissolved inorganic phosphorus (DIP) fluxes averaged 4.5 (±4.6) and 0.16 (±0.17) mmol m−2 d−1, respectively, and compared well with area‐normalized fluxes to Chesapeake Bay. In contrast, SGD‐derived Cu input of 730 (±390) kg yr−1 was a relatively small source of Cu (~3%) to the Elizabeth estuary given that surface water inputs, such as antifouling paints associated with naval operations, are a major component of the Cu budget for this system. These findings were in general agreement with prior studies of SGD for this region.