Nancy E. Kinner

Role of physical heterogeneity in the interpretation of small-scale laboratory and field observations of bacteria, microbial-sized microsphere, and bromide transport through aquifer sediments

The effect of physical variability upon the relative transport behavior of microbial-sized microspheres, indigenous bacteria, and bromide was examined in field and flow-through column studies for a layered, but relatively well sorted, sandy glaciofluvial aquifer. These investigations involved repacked, sieved, and undisturbed aquifer sediments. In the field, peak abundance of labeled bacteria traveling laterally with groundwater flow 6 m downgradient from point of injection was coincident with the retarded peak of carboxylated microspheres (retardation factor, RF = 1.7) at the 8.8 m depth, but preceded the bromide peak and the retarded microsphere peak (RF = 1.5) at the 9.0 m depth. At the 9.5 m depth, the bacterial peak was coincident with both the bromide and the microsphere peaks. Although sorption appeared to be a predominant mechanism responsible for immobilization of microbial-sized microspheres in the aquifer, straining appeared to be primarily responsible for their removal in 0.6-m-long columns of repacked, unsieved aquifer sediments. The manner in which the columns were packed also affected optimal size for microsphere transport, which in one experiment was near the size of the small (∼2 μm) groundwater protozoa (flagellates). These data suggest that variability in aquifer sediment structure can be important in interpretation of both small-scale field and laboratory experiments examining microbial transport behavior.

Protists from a sewage‐contaminated aquifer on cape cod, Massachusetts

Several species of flagellates (genera Bodo, Cercomonas, Cryptaulax, Cyathomonas, Goniomonas, Spumella) have been identified in cultures from a plume of organic contamination (treated sewage effluent) within an aquifer on Cape Cod, Massachusetts. Amoebae and numerous unidentifiable 2‐ to 3‐μm flagellates have also been observed. As a rule, flagellates were associated with solid surfaces, or were capable of temporary surface attachment, corroborating earlier observations from in situ and column transport experiments suggesting that protists in the Massachusetts aquifer have a high propensity for association with sediment grain surfaces. Based on the fact that cultures from the uncontaminated part of the aquifer yielded only a few species of protists, it is hypothesized that the greater abundance and variety of food sources in the contaminant plume is capable of supporting a greater number of protistan species.

Unexpected Sink for Deepwater Horizon Oil May Influence Future Spill Response

A town hall meeting was organized by the Marine Oil Snow Sedimentation and Flocculent Accumulation (MOSSFA) inter-consortia Gulf of Mexico Research Initiative (GoMRI) working group and the Center for Spills in the Environment in conjunction with the Gulf of Mexico Oil Spill and Ecosystem Science Conference. The meeting had the goal of evaluating sedimentation to the seafloor as a significant pathway and fate of oil after the Deepwater Horizon (DwH) well blowout in 2010. About 78,000 cubic meters of crude oil were released into the Gulf of Mexico from a depth of 1500 meters for 86 days, spreading over a large area. Natural and chemically enhanced dispersion, evaporation, dissolution, burning, surface skimming, and direct capture at the wellhead accounted for a significant proportion of the released oil, but the fate of at least 30% of the oil remains unknown. Scientists from different research consortia studying sediments and marine snow in the Gulf began to observe signs of increased sedimentation and hydrocarbon deposition. Sediment mass accumulation rates for the northern Gulf of Mexico increased sixfold to eightfold in 2010, directly following the DwH blowout.

Effect of Growth Conditions and Staining Procedure upon the Subsurface Transport and Attachment Behaviors of a Groundwater Protist

ABSTRACT The transport and attachment behaviors of Spumella guttula (Kent), a nanoflagellate (protist) found in contaminated and uncontaminated aquifer sediments in Cape Cod, Mass., were assessed in flowthrough and static columns and in a field injection-and-recovery transport experiment involving an array of multilevel samplers. Transport of S. guttula harvested from low-nutrient (10 mg of dissolved organic carbon per liter), slightly acidic, granular (porous) growth media was compared to earlier observations involving nanoflagellates grown in a traditional high-nutrient liquid broth. In contrast to the highly retarded (retardation factor of ∼3) subsurface transport previously reported for S. guttula, the peak concentration of porous-medium-grown S. guttula traveled concomitantly with that of a conservative (bromide) tracer. About one-third of the porous-medium-grown nanoflagellates added to the aquifer were transported at least 2.8 m downgradient, compared to only ∼2% of the broth-grown nanoflagellates. Flowthrough column studies revealed that a vital (hydroethidine [HE]) staining procedure resulted in considerably less attachment (more transport) of S. guttula in aquifer sediments than did a staining-and-fixation procedure involving 4′,6′-diamidino-2-phenylindole (DAPI) and glutaraldehyde. The calculated collision efficiency (∼10−2 for porous-medium-grown, DAPI-stained nanoflagellates) was comparable to that observed earlier for the indigenous community of unattached groundwater bacteria that serve as prey. The attachment of HE-labeled S. guttula onto aquifer sediment grains was independent of pH (over the range from pH 3 to 9) suggesting a primary attachment mechanism that may be fundamentally different from that of their prey bacteria, which exhibit sharp decreases in fractional attachment with increasing pH. The high degree of mobility of S. guttula in the aquifer sediments has important ecological implications for the protistan community within the temporally changing plume of organic contaminants in the Cape Cod aquifer.

Protistan predation affects trichloroethene biodegradation in a bedrock aquifer.

ABSTRACT Despite extensive research on the bottom-up force of resource availability (e.g., electron donors and acceptors), slow biodegradation rates and stalling at cis-dichloroethene (cDCE) and vinyl chloride continue to be observed in aquifers contaminated with trichloroethene (TCE). The objective of this research was to gauge the impact of the top-down force of protistan predation on TCE biodegradation in laboratory microcosms. When indigenous bacteria from an electron donor-limited TCE-contaminated bedrock aquifer were present, the indigenous protists inhibited reductive dechlorination altogether. The presence of protists during organic carbon-amended conditions caused the bacteria to elongate (length:width, ≥10:1), but reductive dechlorination was still inhibited. When a commercially available dechlorinating bacterial culture and an organic carbon amendment were added in he presence of protists, the elongated bacteria predominated and reductive dechlorination stalled at cDCE. When protists were removed under organic carbon-amended conditions, reductive dechlorination stalled at cDCE, whereas in the presence organic carbon and bacterial amendments, the total chlorinated ethene concentration decreased, indicating TCE was converted to ethene and/or CO2. The data suggested that indigenous protists grazed dechlorinators to extremely low levels, inhibiting dechlorination altogether. Hence, in situ bioremediation/bioaugmentation may not be successful in mineralizing TCE unless the top-down force of protistan predation is inhibited.

Microbial Processes in Fractured Rock Environments

Little is known about the types and activities of microbes in fractured rock environments, but recent studies in a variety of bedrock formations have documented the presence of a diverse array of prokaryotes (Eubacteria and Archaea) and some protists. The prokaryotes appear to live in both diffusion-dominated microfractures and larger, more conductive open fractures. Some of the prokaryotes are associated with the surfaces of the host rock and mineral precipitates, while other planktonic forms are floating/moving in the groundwater filling the fractures. Studies indicate that the surface-associated and planktonic communities are distinct, and their importance in microbially mediated processes occurring in the bedrock environment may vary, depending on the availability of electron donors/acceptors and nutrients needed by the cells. In general, abundances of microbes are low compared with other environments, because of the paucity of these substances that are transported into the deeper subsurface where most bedrock occurs, unless there is significant pollution with an electron donor. To obtain a complete picture of the microbes present and their metabolic activity, it is usually necessary to sample formation water from specific fractures (versus open boreholes), and fracture surfaces (i.e., cores). Transport of the microbes through the major fracture pathways can be rapid, but may be quite limited in the microfractures. Very low abundances of small (2-3 μm) flagellated protists, which appear to prey upon planktonic bacteria, have been found in a bedrock aquifer. Much more research is needed to expand the understanding of all microbial processes in fractured rock environments.

The Distribution of Protozoa in an Organically Contaminated Aquifer

Protozoa are present in numbers ranging from 104−105/gram dry weight (gdw) in a sandy aquifer contaminated by wastewater effluent. The protozoa are primarily 2-3 urn flagellates. The ratio of free-living bacteria (FLB): protozoa ranges from 100−102. There is no strong relationship between the FLB and protozoa. Studies are being conducted to determine the food source(s) for the protozoa as a means of understanding their role in the biochemical processes occurring within the plume.

Rock Fragments in Trichloroethene Microcosms for Bedrock Aquifers

ABSTRACT The purpose of this study was to evaluate microcosm methods that can be used to predict the in situ anaerobic reductive dechlorination of trichloroethene (TCE) in bedrock aquifers with and without amendments. Microcosm conditions (e.g., surface area:volume ratio, initial TCE concentration [C 0], incubation temperature) were selected to simulate in situ conditions at the site. The effect of rock media in the microcosms as a source of surface area and nutrients was also assessed. Comparison was made between microcosms constructed with sterile rock media and rock media submerged for 45 days in a groundwater well within a TCE plume for field colonization. Statistically significant biotic degradation of TCE was observed in microcosms with both groundwater only and non-field-colonized rock media, when amended with lactate. Measured half-lives were similar to half-lives observed in field conditions. Normalizing biotic results to abiotic results provides a method that deducts the abiotic losses and allows complete comparison between sample rounds, assuming samples were randomly selected for analysis within the sample round to avoid systematic errors or variation.

Microfracture Surface Geochemistry and Adherent Microbial Population Metabolism in TCE-Contaminated Competent Bedrock

A TCE-contaminated competent bedrock site in Portsmouth, NH was used to determine if a relation existed between microfracture surface geochemistry and the ecology and metabolic activity of attached microbes relative to terminal electron accepting processes (TEAPs) and TCE biodegradation. The bedrock is a metasandstone and metashale of the Silurian Kittery Formation. Eleven microfractures (MF 01-11) were extracted from cores of competent rock from 2 boreholes (BBC5 and BBC6) at depths > 21.3 m below ground. The host rock had 3 nominal pore width sizes (131.1, 1.136, and 0.109 μ m), a porosity of 0.8%, and a permeability of < 1 μ d. Microfracture surface precipitates were polycrystalline with grain sizes ranging from 10 to 100 μ m. Petrography and XRD revealed that carbonates and quartz were the dominant microfracture surface precipitates. Mineral distribution was heterogeneous at the 10 μ m scale. Oxidized and reduced iron species were identified with XPS on the microfracture precipitate surfaces. Carbon functional groups characteristic of NOM were also identified. SIMS mass fragment fingerprints suggested that TCE, PCE and/or VC were possibly adsorbed to NOM on the microfracture surfaces. Packer waters were alkaline (131–190 mg/L as CaCO3, pH 8.8 to 9.6), mildly reducing (Eh of −208 to 160 mV, DO of 0.4 to 2.5 mg/L), with low NPDOC values (0.8–1.7 mg/L), and measurable Fe (II) (0.1 mg/L) and Fe (III) (0.02 to 0.3 mg/L). Sulfate was the dominant anion in the packer sample water (110–120 mg/L). No sulfide was detected. H 2 was present in a number of the BBC wells at the site (2.2–7.3 nM). Amplification with specific primer sets of seven microfractures from BBC5 showed the presence of bacteria, Archaea, anaerobic dehalorespirers (Dehalococcoides sp.), sulfate reducing bacteria, and iron reducing bacteria (Geobacteraceae). Redox zonation may exist relative to spatial distance from within the microfracture network to the open fracture system. The microfracture surface precipitates, frequently spatially complex and comprised of a variety of C-, Fe- and S-containing minerals, may be another region for redox zonation. Fe was the dominant microfracture surface element and active Fe cycling is suspected. However, the primer data suggest that the microfracture network may have been more reducing than the open fracture system. In this case, the microfracture network may constitute a zone where more reductive metabolic processes occur, making this system similar to biogeochemical redox zones found in other environments.

RESEARCH NEEDS IN OIL SPILL RESPONSE

ABSTRACT As funding for spill research and development (R&D) has declined in recent years, partnerships among relevant federal and state agencies, industry and academia have increased in importance...