Thomas J. Boyd

Dissolved oxygen saturation controls PAH biodegradation in freshwater estuary sediments.

Polycyclic aromatic hydrocarbons (PAHs) are common contaminants in terrestrial and aquatic environments and can represent a significant constituent of the carbon pool in coastal sediments. We report here the results of an 18-month seasonal study of PAH biodegradation and heterotrophic bacterial production and their controlling biogeochemical factors from 186 sediment samples taken in a tidally influenced freshwater estuary. For each sampling event, measurements were averaged from 25–45 stations covering ∼250 km2. There was a clear relationship between bacterial production and ambient temperature, but none between production and bottom water dissolved oxygen (DO) % saturation or PAH concentrations. In contrast with other studies, we found no effect of temperature on the biodegradation of naphthalene, phenanthrene, or fluoranthene. PAH mineralization correlated with bottom water DO saturation above 70% (r2 > 0.99). These results suggest that the proportional utilization of PAH carbon to natural organic carbon is as much as three orders of magnitude higher during cooler months, when water temperatures are lower and DO % saturation is higher. Infusion of cooler, well-oxygenated water to the water column overlying contaminated sediments during the summer months may stimulate PAH metabolism preferentially over non-PAH organic matter.

Transport, Deposition and Biodegradation of Particle Bound Polycyclic Aromatic Hydrocarbons in a Tidal Basin of an Industrial Watershed

Polycylic aromatic hydrocarbons (PAHs) are commoncontaminants in industrial watersheds. Their origin,transport and fate are important to scientists,environmental managers and citizens. The Philadelphia NavalReserve Basin (RB) is a small semi-enclosed embayment nearthe confluence of the Schuylkill and Delaware Rivers inPennsylvania (USA). We conducted a study at this site todetermine the tidal flux of particles and particle-boundcontaminants associated with the RB. Particle traps wereplaced at the mouth and inside the RB and in the Schuylkilland Delaware Rivers. There was net particle deposition intothe RB, which was determined for three seasons. Spring andfall depositions were highest (1740 and 1230 kg ofparticles, respectively) while winter deposition wasinsignificant. PAH concentrations on settling particlesindicated a net deposition of 12.7 g PAH in fall and 2.1 gPAH in spring over one tidal cycle. There was nosignificant PAH deposition in the winter. Biodegradationrates, calculated from 14C-labeled PAH substratemineralization, could attenuate only about 0.25% of the PAHdeposited during a tidal cycle in fall. However, in thespring, biodegradation could be responsible for degrading50% of the settling PAHs. The RB appears to be a sink forPAHs in this watershed.

Optical Proxies for Terrestrial Dissolved Organic Matter in Estuaries and Coastal Waters

Optical proxies, especially DOM fluorescence, were used to track terrestrial DOM fluxes through estuaries and coastal waters by comparing models developed for several coastal ecosystems. Key to using optical properties is validating and calibrating them with chemical measurements, such as lignin-derived phenols - a proxy to quantify terrestrial DOM. Utilizing parallel factor analysis (PARAFAC), and comparing models statistically using the OpenFluor database (http://www.openfluor.org) we have found common, ubiquitous fluorescing components which correlate most strongly with lignin phenol concentrations in several estuarine and coastal environments. Optical proxies for lignin were computed for the following regions: Mackenzie River Estuary, Atchafalaya River Estuary, Charleston Harbor, Chesapeake Bay, and Neuse River Estuary. The slope of linear regression models relating CDOM absorption at 350 nm (a350) to DOC and to lignin, varied 5 to 10 fold among systems. Where seasonal observations were available from a region, there were distinct seasonal differences in equation parameters for these optical proxies. Despite variability, overall models using single linear regression were developed that related dissolved organic carbon (DOC) concentration to CDOM (DOC = 40×a350+138; R2 = 0.77; N = 130) and lignin (Σ8) to CDOM (Σ8 = 2.03×a350-0.5; R2 = 0.87; N = 130). This wide variability suggested that local or regional optical models should be developed for predicting terrestrial DOM flux into coastal oceans and taken into account when upscaling to remote sensing observations and calibrations.

2,4,6-Trinitrotoluene mineralization and bacterial production rates of natural microbial assemblages from coastal sediments.

The nitrogenous energetic constituent, 2,4,6-Trinitrotoluene (TNT), is widely reported to be resistant to bacterial mineralization (conversion to CO(2)); however, these studies primarily involve bacterial isolates from freshwater where bacterial production is typically limited by phosphorus. This study involved six surveys of coastal waters adjacent to three biome types: temperate broadleaf, northern coniferous, and tropical. Capacity to catabolize and mineralize TNT ring carbon to CO(2) was a common feature of natural sediment assemblages from these coastal environments (ranging to 270+/-38 μg C kg(-1) d(-1)). More importantly, these mineralization rates comprised a significant proportion of total heterotrophic production. The finding that most natural assemblages surveyed from these ecosystems can mineralize TNT ring carbon to CO(2) is consistent with recent reports that assemblage components can incorporate TNT ring carbon into bacterial biomass. These data counter the widely held contention that TNT is recalcitrant to bacterial catabolism of the ring carbon in natural environments.

Effects of Dissolved and Complexed Copper on Heterotrophic Bacterial Production in San Diego Bay

Bacterial abundance and production, free (uncomplexed) copper ion concentration, total dissolved copper concentration, dissolved organic carbon (DOC), total suspended solids (TSS), and chlorophyll a were measured over the course of 1 year in a series of 27 sample “Boxes” established within San Diego Bay. Water was collected through a trace metal-clean system so that each Box’s sample was a composite of all the surface water in that Box. Bacterial production, chlorophyll a, TSS, DOC, and dissolved copper all generally increased from Box 1 at the mouth of the Bay to Box 27 in the South or back Bay. Free copper ion concentration generally decreased from Box 1 to Box 27 presumably due to increasing complexation capacity within natural waters. Based on correlations between TSS, chlorophyll a, bacterial production or DOC and the ratio of dissolved to free Cu ion, both DOC and particulate (bacteria and algae) fractions were potentially responsible for copper complexation, each at different times of the year. CuCl2 was added to bacterial production assays from 0 to 10 μg L−1 to assess acute copper toxicity to the natural microbial assemblage. Interestingly, copper toxicity appeared to increase with decreases in free copper from the mouth of the Bay to the back Bay. This contrasts the free-ion activity model in which higher complexation capacity should afford greater copper protection. When cell-specific growth rates were calculated, faster growing bacteria (i.e. toward the back Bay) appeared to be more susceptible to free copper toxicity. The protecting effect of natural dissolved organic material (DOM) concentrated by tangential flow ultrafiltration (>1 kDa), illite and kaolinite minerals, and glutathione (a metal chelator excreted by algae under copper stress) was assessed in bacterial production assays. Only DOM concentrate offered any significant protection to bacterial production under increased copper concentrations. Although the potential copper protecting agents were allowed to interact with added copper before natural bacteria were added to production assays, there may be a temporal dose–response relationship that accounts for higher toxicity in short production assays. Regardless, it appears that effective natural complexation of copper in the back portions of San Diego Bay limits exposure of native bacterial assemblages to free copper ion, resulting in higher bacterial production.

PAH mineralization and bacterial organotolerance in surface sediments of the Charleston Harbor estuary

Semi-volatile organic compounds (SVOCs) in estuarine waters can adversely affect biota but watershed sources can be difficult to identify because these compounds are transient. Natural bacterial assemblages may respond to chronic, episodic exposure to SVOCs through selection of more organotolerant bacterial communities. We measured bacterial production, organotolerance and polycyclic aromatic hydrocarbon (PAH) mineralization in Charleston Harbor and compared surface sediment from stations near a known, permitted SVOC outfall (pulp mill effluent) to that from more pristine stations. Naphthalene additions inhibited an average of 77% of bacterial metabolism in sediments from the more pristine site (Wando River). Production in sediments nearest the outfall was only inhibited an average of 9% and in some cases, was actually stimulated. In general, the stations with the highest rates of bacterial production also were among those with the highest rates of PAH mineralization. This suggests that the capacity to mineralize PAH carbon is a common feature amongst the bacterial assemblage in these estuarine sediments and could account for an average of 5.6% of bacterial carbon demand (in terms of production) in the summer, 3.3% in the spring (April) and only 1.2% in winter (December).

Radiocarbon and Stable Carbon Isotope Analysis to Confirm Petroleum Natural Attenuation in the Vadose Zone

CO2 and CH4 radiocarbon and stable carbon isotope ratios were used to assess natural attenuation at a fuel-contaminated soil site at the Norfolk Navy Base, Norfolk, VA (USA). Soil gas samples were collected spatially over a monitoring network in October 2002 and in March 2003. CO2 and CH4 from regions with high petroleum concentrations were 14C-depleted relative to uncontaminated areas. 14C-depleted methane suggested methanogenic hydrocarbon degradation. The difference in CO2 age between background and plume-influenced areas indicated that approximately 90% of the CO2 at the latter was petroleum derived, making contaminant the primary source of carbon for the microbial assemblage.

Effects of oxygenation on hydrocarbon biodegradation in a hypoxic environment

In 1984, an underground storage tank leaked approximately 41,000 L of gasoline into the ground water at the Naval Construction Battalion Command in Port Hueneme, CA (USA). Benzene, toluene, ethylbenzene, and xylenes (BTEX) contamination stimulated remedial action. In 1995, a ground water circulation well (GCW) and network of surrounding monitoring wells were installed. After year of operation, dissolved oxygen and nitrate concentrations remained low in all monitoring wells. Benzene utilization (the sum of respiration, uptake, and conversion to polar compounds) ranged from 0.03 to 4.6 µg L−1 h−1, and toluene utilization ranged from 0.01 to 5.2 µg L−1 h−1. Heterotrophic bacterial productivity (total carbon assimilation) increased dramatically in the GCW, although benzene and toluene utilization decreased markedly relative to surrounding wells. Benzene and toluene uptake accounted for a significant proportion (mean=22%) of the heterotrophic bacterial productivity except within the GCW, indicating other fuel contaminant or indigenous organic carbon and not BTEX compounds served as primary carbon source. The GCW effectively air-stripped BTEX compounds, but failed to stimulate benzene and toluene biodegradation and thus would not be effective for stimulating BTEX bioremediation under current deployment parameters. Air stripping was three orders of magnitude more effective than biodegradation for removing benzene and toluene in the GCW.

Use of stable carbon isotopes and multivariate statistics to source-apportion fuel hydrocarbons

Differences in hydrocarbon samples were assessed using compound-specific isotope analysis (CSIA). Samples from on-water spills and fuel tanks from possible sources (moored ships) were taken from the Norfolk Navy Base in Norfolk, VA (USA) and volatile organic compound (VOC) samples were taken from a multiple leaking underground fuel tank site in Hazleton, PA (USA). Individual compound stable carbon isotope ratios were measured and the data subjected to several multivariate statistical tests to determine similarity between sample hydrocarbon source(s). Multiple analysis of variance, principal components analysis and hierarchical clustering showed similarities between discreet samples at both sites and in the case of the Norfolk samples confirmed a shipboard leak. CSIA coupled with multivariate statistics was found to be a useful tool for source-apportioning hydrocarbons.

Radiocarbon-depleted CO2 evidence for fuel biodegradation at the Naval Air Station North Island (USA) fuel farm site

Dissolved CO(2) radiocarbon and stable carbon isotope ratios were measured in groundwater from a fuel contaminated site at the North Island Naval Air Station in San Diego, CA (USA). A background groundwater sampling well and 16 wells in the underground fuel contamination zone were evaluated. For each sample, a two end-member isotopic mixing model was used to determine the fraction of CO(2) derived from fossil fuel. The CO(2) fraction from fossil sources ranged from 8 to 93% at the fuel contaminated site, while stable carbon isotope values ranged from -14 to +5‰VPDB. Wells associated with highest historical and contemporary fuel contamination showed the highest fraction of CO(2) derived from petroleum (fossil) sources. Stable carbon isotope ratios indicated sub-regions on-site with recycled CO(2) (δ(13)CO(2) as high as +5‰VPDB) - most likely resulting from methanogenesis. Ancillary measurements (pH and cations) were used to determine that no fossil CaCO(3), for instance limestone, biased the analytical conclusions. Radiocarbon analysis is verified as a viable and definitive technique for confirming fossil hydrocarbon conversion to CO(2) (complete oxidation) at hydrocarbon-contaminated groundwater sites. The technique should also be very useful for assessing the efficacy of engineered remediation efforts and by using CO(2) production rates, contaminant mass conversion over time and per unit volume.