Brian J. Mailloux

Hydrogeological characterization of the south oyster bacterial transport site using geophysical data

A multidisciplinary research team has conducted a field-scale bacterial transport study within an uncontaminated sandy Pleistocene aquifer near Oyster, Virginia. The overall goal of the project was to evaluate the importance of heterogeneities in controlling the field-scale transport of bacteria that are injected into the ground for remediation purposes. Geochemical, hydrological, geological, and geophysical data were collected to characterize the site prior to conducting chemical and bacterial injection experiments. In this paper we focus on results of a hydrogeological characterization effort using geophysical data collected across a range of spatial scales. The geophysical data employed include surface ground-penetrating radar, radar cross-hole tomography, seismic cross-hole tomography, cone penetrometer, and borehole electromagnetic flowmeter. These data were used to interpret the subregional and local stratigraphy, to provide high-resolution hydraulic conductivity estimates, and to provide information about the log conductivity spatial correlation function. The information from geophysical data was used to guide and assist the field operations and to constrain the numerical bacterial transport model. Although more field work of this nature is necessary to validate the usefulness and cost-effectiveness of including geophysical data in the characterization effort, qualitative and quantitative comparisons between tomographically obtained flow and transport parameter estimates with hydraulic well bore and bromide breakthrough measurements suggest that geophysical data can provide valuable, high-resolution information. This information, traditionally only partially obtainable by performing extensive and intrusive well bore sampling, may help to reduce the ambiguity associated with hydrogeological heterogeneity that is often encountered when interpreting field-scale bacterial transport data.

Development of a vital fluorescent staining method for monitoring bacterial transport in subsurface environments.

ABSTRACT Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate,Comamonas sp. strain DA001. Potential compounds were first screened to determine staining efficiencies and adverse side effects. 5-(And 6-)-carboxyfluorescein diacetate, succinimidyl ester (CFDA/SE) efficiently stained DA001 without causing undesirable effects on cell adhesion or viability. Members of many other gram-negative and gram-positive bacterial genera were also effectively stained with CFDA/SE. More than 95% of CFDA/SE-stained Comamonas sp. strain DA001 cells incubated in artificial groundwater (under no-growth conditions) remained fluorescent for at least 28 days as determined by epifluorescent microscopy and flow cytometry. No differences in the survival and culturability of CFDA/SE-stained and unstained DA001 cells in groundwater or saturated sediment microcosms were detected. The bright, yellow-green cells were readily distinguished from autofluorescing sediment particles by epifluorescence microscopy. A high throughput method using microplate spectrofluorometry was developed, which had a detection limit of mid-105CFDA-stained cells/ml; the detection limit for flow cytometry was on the order of 1,000 cells/ml. The results of laboratory-scale bacterial transport experiments performed with intact sediment cores and nondividing DA001 cells revealed good agreement between the aqueous cell concentrations determined by the microplate assay and those determined by other enumeration methods. This research indicates that CFDA/SE is very efficient for labeling cells for bacterial transport experiments and that it may be useful for other microbial ecology research as well.

Theoretical prediction of collision efficiency between adhesion-deficient bacteria and sediment grain surface

Abstract Our earlier results concerning bacterial transport of an adhesion-deficient strain Comamonas sp. (DA001) in intact sediment cores from near South Oyster, VA demonstrated that grain size is the principle factor controlling bacterial retention, and that Fe and Al hydroxide mineral coatings are of secondary importance. The experimentally determined collision efficiency ( α ) was in the range of 0.003–0.026 and did not correlate with the Fe and Al concentration. This study attempts to theoretically predict α , and identifies factors responsible for the observed low α . The modified Derjaguin–Landau–Verwey–Overbeek (DLVO) theory was used to calculate the total intersurface potential energy as a function of separation distance between bacterial and sediment surfaces and to provide insights into the relative importance of bacterial and sediment grain surface properties in controlling magnitude of α . Different models for calculating theoretical α were developed and compared. By comparing theoretical α values from different models with previously published experimental α values, it is possible to identify a suitable model for predicting α . When DA001 bacteria interact with quartz surfaces, the theoretical α best predicts experimental α when DA001 cells are reversibly attached to the secondary minimum of the energy interaction curve and α depends on the probability of escape from that energy well. No energy barrier opposes bacterial attachment to clean iron oxide surface of positive charge at sub-neutral pH, thus the model predicts α of unity. When the iron oxide is equilibrated with natural groundwater containing 5–10 ppm of dissolved organic carbon (DOC), its surface charge reverses, and the model predicts α to be on the order of 0.2. The theoretical α for DA001 in the natural sediments from South Oyster, VA was estimated by representing the surface potential of the sediment as a patch-wise binary mixture of negatively charged quartz ( ζ =−60 mV) and organic carbon coated Fe–Al hydroxides ( ζ =−2 mV). Such a binary mixing approach generates α that closely matches the experimental α . This study demonstrates that it is possible to predict α from known bacterial and grain surface properties.

Extended tailing of bacteria following breakthrough at the Narrow Channel Focus Area, Oyster, Virginia

Extended tailing of low bacterial concentrations following breakthrough at the Narrow Channel focus area was observed for 4 months. Bacterial attachment and detachment kinetics associated with breakthrough and extended tailing were determined by fitting a one-dimensional transport model to the field breakthrough-tailing data. Spatial variations in attachment rate coefficient (kf) were observed under forced gradient conditions (i.e., k f decreased as travel distance increased), possibly because of decreased bacterial adhesion with increased transport distance. When pore water velocity decreased by an order of magnitude at 9 days following injection, apparent bacterial attachment rate coefficients did not decrease with velocity as expected from filtration theory, but, instead, increased greatly for most of the wells. The coincidence of the increase in apparent attachment rate coefficient with the occurrence of protist blooms suggested that the loss of bacteria from the aqueous phase during the protist blooms was not governed by filtration but rather was governed by predation. Simulations were performed to examine the transport distances achieved with and without detachment, using attachment and detachment rate coefficients similar to those obtained in this field study. Simulations that included detachment showed that transport distances of bacteria may significantly increase because of detachment under the conditions examined.

Microbial mineral weathering for nutrient acquisition releases arsenic.

ABSTRACT Tens of millions of people in Southeast Asia drink groundwater contaminated with naturally occurring arsenic. How arsenic is released from the sediment into the water remains poorly understood. Here, we show in laboratory experiments that phosphate-limited cells of Burkholderia fungorum mobilize ancillary arsenic from apatite. We hypothesize that arsenic mobilization is a by-product of mineral weathering for nutrient acquisition. The released arsenic does not undergo a redox transformation but appears to be solubilized from the apatite mineral lattice during weathering. Analysis of apatite from the source area in the Himalayan basin indicates the presence of elevated levels of arsenic, with an average concentration of 210 mg/kg. The rate of arsenic release is independent of the initial dissolved arsenic concentration and occurs at phosphate levels observed in Bangladesh aquifers. We also demonstrate the presence of the microbial phenotype that releases arsenic from apatite in Bangladesh aquifer sediments and groundwater. These results suggest that microbial mineral weathering for nutrient acquisition could be an important mechanism for arsenic mobilization.

Comparison of fecal indicators with pathogenic bacteria and rotavirus in groundwater

Groundwater is routinely analyzed for fecal indicators but direct comparisons of fecal indicators to the presence of bacterial and viral pathogens are rare. This study was conducted in rural Bangladesh where the human population density is high, sanitation is poor, and groundwater pumped from shallow tubewells is often contaminated with fecal bacteria. Five indicator microorganisms (E. coli, total coliform, F+RNA coliphage, Bacteroides and human-associated Bacteroides) and various environmental parameters were compared to the direct detection of waterborne pathogens by quantitative PCR in groundwater pumped from 50 tubewells. Rotavirus was detected in groundwater filtrate from the largest proportion of tubewells (40%), followed by Shigella (10%), Vibrio (10%), and pathogenic E. coli (8%). Spearman rank correlations and sensitivity-specificity calculations indicate that some, but not all, combinations of indicators and environmental parameters can predict the presence of pathogens. Culture-dependent fecal indicator bacteria measured on a single date did not predict total bacterial pathogens, but annually averaged monthly measurements of culturable E. coli did improve prediction for total bacterial pathogens. A qPCR-based E. coli assay was the best indicator for the bacterial pathogens. F+RNA coliphage were neither correlated nor sufficiently sensitive towards rotavirus, but were predictive of bacterial pathogens. Since groundwater cannot be excluded as a significant source of diarrheal disease in Bangladesh and neighboring countries with similar characteristics, the need to develop more effective methods for screening tubewells with respect to microbial contamination is necessary.

Fecal Contamination of Shallow Tubewells in Bangladesh Inversely Related to Arsenic

The health risks of As exposure due to the installation of millions of shallow tubewells in the Bengal Basin are known, but fecal contamination of shallow aquifers has not systematically been examined. This could be a source of concern in densely populated areas with poor sanitation because the hydraulic travel time from surface water bodies to shallow wells that are low in As was previously shown to be considerably shorter than for shallow wells that are high in As. In this study, 125 tubewells 6−36 m deep were sampled in duplicate for 18 months to quantify the presence of the fecal indicator Escherichia coli. On any given month, E. coli was detected at levels exceeding 1 most probable number per 100 mL in 19−64% of all shallow tubewells, with a higher proportion typically following periods of heavy rainfall. The frequency of E. coli detection averaged over a year was found to increase with population surrounding a well and decrease with the As content of a well, most likely because of downward transport of E. coli associated with local recharge. The health implications of higher fecal contamination of shallow tubewells, to which millions of households in Bangladesh have switched in order to reduce their exposure to As, need to be evaluated.

Advection of surface-derived organic carbon fuels microbial reduction in Bangladesh groundwater

Chronic exposure to arsenic (As) by drinking shallow groundwater causes widespread disease in Bangladesh and neighboring countries. The release of As naturally present in sediment to groundwater has been linked to the reductive dissolution of iron oxides coupled to the microbial respiration of organic carbon (OC). The source of OC driving this microbial reduction—carbon deposited with the sediments or exogenous carbon transported by groundwater—is still debated despite its importance in regulating aquifer redox status and groundwater As levels. Here, we used the radiocarbon (14C) signature of microbial DNA isolated from groundwater samples to determine the relative importance of surface and sediment-derived OC. Three DNA samples collected from the shallow, high-As aquifer and one sample from the underlying, low-As aquifer were consistently younger than the total sediment carbon, by as much as several thousand years. This difference and the dominance of heterotrophic microorganisms implies that younger, surface-derived OC is advected within the aquifer, albeit more slowly than groundwater, and represents a critical pool of OC for aquifer microbial communities. The vertical profile shows that downward transport of dissolved OC is occurring on anthropogenic timescales, but bomb 14C-labeled dissolved OC has not yet accumulated in DNA and is not fueling reduction. These results indicate that advected OC controls aquifer redox status and confirm that As release is a natural process that predates human perturbations to groundwater flow. Anthropogenic perturbations, however, could affect groundwater redox conditions and As levels in the future.

Tectonic controls on the hydrogeology of the Rio Grande Rift, New Mexico

Mathematical modeling is used in this study to assess how tectonic movement of fault blocks and fault permeability influence the present-day and paleohydrogeology of the Rio Grande Rift near Socorro, New Mexico. Our analysis focuses on active and ancient groundwater flow patterns and hot spring development within the southern La Jencia and Socorro subbasins. The best agreement between model results and present-day and paleoheat flow data was achieved by representing faults as passive surfaces and incorporating 2 km of moderately permeable (10−14.0 m2) fractured crystalline rocks into the hydrogeologic model. Quantitative results indicate that changes in groundwater flow patterns across the basin are primarily generated by the truncation/reconnection of aquifers and confining units. Calculated flow patterns help to explain the annealing of apatite fission tracks within Eocene Baca Formation clasts to the east of Socorro, potassium metasomatism mass balance constraints within Oligocene volcanics and overlying Santa Fe Group deposits, and the timing of barite/fluorite ore mineralization within the Gonzales prospect on the eastern edge of the Rio Grande Rift. We estimate that about 5% of mountain front recharge penetrates to a depth of 2.8 km below the sedimentary pile. This may have implications for water resource planners who have historically focused on groundwater resource development within the shallow alluvial deposits along the Rio Grande Rift valley.

Redox trapping of arsenic during groundwater discharge in sediments from the Meghna riverbank in Bangladesh

Groundwater arsenic (As) is elevated in the shallow Holocene aquifers of Bangladesh. In the dry season, the shallow groundwater discharges to major rivers. This process may influence the chemistry of the river and the hyporheic zone sediment. To assess the fate of As during discharge, surface (0–5 cm) and subsurface (1–3 m) sediment samples were collected at 9 sites from the bank of the Meghna River along a transect from its northern source (25° N) to the Bay of Bengal (22.5° N). Bulk As concentrations of surface sediment averaged 16 ± 7 mg/kg (n = 9). Subsurface sediment contained higher mean concentrations of As of 4,000 mg/kg (n = 14), ranging from 1 to 23,000 mg/kg As, with >100 mg/kg As measured at 8 sites. X-ray absorption near-edge structure spectroscopy indicated that As was mainly arsenate and arsenite, not As-bearing sulfides. We hypothesize that the elevated sediment As concentrations form as As-rich groundwater discharges to the river, and enters a more oxidizing environment. A significant portion of dissolved As sorbs to iron-bearing minerals, which form a natural reactive barrier. Recycling of this sediment-bound As to the Ganges-Brahmaputra-Meghna Delta aquifer provides a potential source of As to further contaminate groundwater. Furthermore, chemical fluxes from groundwater discharge from the Ganges-Brahmaputra-Meghna Delta may be less than previous estimates because this barrier can immobilize many elements.