Abstract. 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 highresolution 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.

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.

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 ( 14 C) 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 14 C-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.A quifer redox status is a major factor affecting groundwater composition and its suitability for human consumption. The most egregious example is the dire health impact of elevated levels of arsenic (As) in groundwater drawn with inexpensive handpumped tube wells by more than 100 million villagers across South, Southeast, and East Asia (1, 2). Aquifer redox status is largely controlled by microbial respiration of organic carbon (OC) coupled to the utilization of terminal electron acceptors. Reduction of iron (Fe) oxides containing As coupled with the oxidation of OC is the dominant process causing the accumulation of As in groundwater (1), but there is no consensus on the source of OC that fuels these transformations. Such information is critical to understanding subsurface carbon cycling, redox reactions, and the vulnerability of groundwater aquifers to perturbation.Human perturbations have long been suggested to exacerbate the problem of elevated As in groundwater in several ways. Given the connectivity between contaminated surface waters and aquifers, widespread pumping could redistribute As from the surface to depth (3, 4) or between aquifers (5). This pumping could potentially also deliver reactive OC from ponds and latrines to depth, causing microbial reduction and As release (3, 4). Fluorescent spectra of dissolved organic carbon (DOC), correlations between sedimentary ...

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/SEstained 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-10 5 CFDA-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.

Whereas serious health consequences of widespread consumption of groundwater elevated in As have been documented in several South Asian countries, the mechanisms responsible for As mobilization in reducing aquifers remain poorly understood. We document here a previously unrecognized and consistent relationship between dissolved As concentrations in reducing groundwater and the phosphate-mobilizable As content of aquifer sediment for a set of precisely depth-matched samples from across Bangladesh. The relationship holds across nearly 3 orders of magnitude in As concentrations and suggests that regional as well as local patterns of dissolved As in shallow groundwater are set by the solid phase according to a remarkably constant ratio of ∼250 μg/L dissolved As per 1 mg/kg P-mobilizable As. We use this relationship in a simple model of groundwater recharge to propose that the distribution of groundwater As in shallow aquifers of the Bengal Basin could primarily reflect the different flushing histories of sand formations deposited in the region over the past several thousand years.