David W. Metge

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.

What Makes a Natural Clay Antibacterial

Natural clays have been used in ancient and modern medicine, but the mechanism(s) that make certain clays lethal against bacterial pathogens has not been identified. We have compared the depositional environments, mineralogies, and chemistries of clays that exhibit antibacterial effects on a broad spectrum of human pathogens including antibiotic resistant strains. Natural antibacterial clays contain nanoscale (<200 nm), illite-smectite and reduced iron phases. The role of clay minerals in the bactericidal process is to buffer the aqueous pH and oxidation state to conditions that promote Fe(2+) solubility. Chemical analyses of E. coli killed by aqueous leachates of an antibacterial clay show that intracellular concentrations of Fe and P are elevated relative to controls. Phosphorus uptake by the cells supports a regulatory role of polyphosphate or phospholipids in controlling Fe(2+). Fenton reaction products can degrade critical cell components, but we deduce that extracellular processes do not cause cell death. Rather, Fe(2+) overwhelms outer membrane regulatory proteins and is oxidized when it enters the cell, precipitating Fe(3+) and producing lethal hydroxyl radicals.

Influence of organic matter on the transport of Cryptosporidium parvum oocysts in a ferric oxyhydroxide-coated quartz sand saturated porous medium

To assess the effect of organic matter on the transport of Cryptosporidium parvum oocysts in a geochemically heterogeneous saturated porous medium, we measured the breakthrough and collision efficiencies of oocysts as a function of dissolved organic matter concentration in a flow-through column containing ferric oxyhydroxide-coated sand. We characterized the surface properties of the oocysts and ferric oxyhydroxide-coated sand using microelectrophoresis and streaming potential, respectively, and the amount of organic matter adsorbed on the ferric oxyhydroxide-coated sand as a function of the concentration of dissolved organic matter (a fulvic acid isolated from Florida Everglades water). The dissolved organic matter had no significant effect on the zeta potential of the oocysts. Low concentrations of dissolved organic matter were responsible for reversing the charge of the ferric oxyhydroxide-coated sand surface from positive to negative. The charge reversal and accumulation of negative charge on the ferric oxyhydroxide-coated sand led to increases in oocyst breakthrough and decreases in oocyst collision efficiency with increasing dissolved organic matter concentration. The increase in dissolved organic matter concentration from 0 to 20mg L(-1) resulted in a two-fold decrease in the collision efficiency.

Assessing the vulnerability of a municipal well field to contamination in a karst aquifer

Proposed expansion of extractive lime-rock mines near the Miami–Dade County Northwest well field and Everglades wetland areas has garnered intense scrutiny by government, public, environmental stakeholders, and the media because of concern that mining will increase the risk of pathogen contamination. Rock mines are excavated to the same depth as the well fields primary producing zone. The underlying karst Biscayne aquifer is a triple-porosity system characterized by (1) a matrix of interparticle porosity and separate vug porosity; (2) touching-vug porosity that forms preferred, stratiform passageways; and, less commonly, (3) conduit porosity formed by thin solution pipes, bedding-plane vugs, and cavernous vugs. Existing ground-water flow and particle tracking models do not provide adequate information regarding the ability of the aquifer to limit the advective movement of pathogens and other contaminants. Chemical transport and colloidal mobility properties have been delineated using conservative and microsphere-surrogate tracers for Cryptosporidium parvum . Forced-gradient tests were executed by introducing conservative tracers into injection wells located 100 m (328 ft) from a municipal-supply well. Apparent mean advective velocity between the wells is one to two orders of magnitude greater than previously measured. Touching-vug, stratiform flow zones are efficient pathways for tracer movement at the well field. The effective porosity for a continuum model between the point of injection and tracer recovery ranges from 2 to 4 percent and is an order of magnitude smaller than previously assumed. Existing well-field protection zones were established using porosity estimates based on specific yield. The effective, or kinematic, porosity of a Biscayne aquifer continuum model is lower than the total porosity, because high velocities occur along preferential flow paths that result in faster times of travel than can be represented with the ground-water flow equation. Tracer tests indicate that the relative ease of contaminant movement to municipal supply wells is much greater than previously considered.

Pathogen and chemical transport in the karst limestone of the Biscayne aquifer: 1. Revised conceptualization of groundwater flow

[1] The Biscayne aquifer is a highly transmissive karst limestone that serves as the sole source of drinking water to over two million residents in south Florida. The aquifer is characterized by eogenetic karst, where the most transmissive void space can be an interconnected, touching-vug, biogenically influenced porosity of biogenic origin. Public supply wells in the aquifer are in close proximity to lakes established by surface mining. The mining of the limestone has occurred to the same depths as the production wells, which has raised concerns about pathogen and chemical transport from these surface water bodies. Hydraulic and forced gradient tracer tests were conducted to augment geologic and geophysical studies and to develop a hydrogeologic conceptual model of groundwater flow and chemical transport in the Biscayne aquifer. Geologic and geophysical data indicate multiple, areally extensive subhorizontal preferential flow zones of vuggy limestone separated by rock with a matrix pore system. The hydraulic response from an aquifer test suggests that the Biscayne aquifer behaves as a dual-porosity medium; however, the results of the tracer test showed rapid transport similar to other types of karst. The tracer test and concurrent temperature logging revealed that only one of the touching-vug flow zones dominates transport near the production wells. On the basis of the rising limb of the breakthrough curve, the dispersivity is estimated to be less than 3% of the tracer travel distance, which suggests that the fastest flow paths in the formation are likely to yield limited dilution of chemical constituents.

Effects on Groundwater Microbial Communities of an Engineered 30-Day In Situ Exposure to the Antibiotic Sulfamethoxazole

Effects upon microbial communities from environmental exposure to concentrations of antibiotics in the μg L(-1) range remain poorly understood. Microbial communities from an oligotrophic aquifer (estimated doubling rates of only once per week) that were previously acclimated (AC) or unacclimated (UAC) to historical sulfamethoxazole (SMX) contamination, and a laboratory-grown Pseudomonas stutzeri strain, were exposed to 240-520 μg L(-1) SMX for 30 days in situ using filter chambers allowing exposure to ambient groundwater, but not to ambient microorganisms. SMX-exposed UAC bacterial communities displayed the greatest mortality and impairment (viable stain assays), the greatest change in sensitivity to SMX (dose-response assays), and the greatest change in community composition (Terminal Restriction Fragment Length Polymorphism; T-RFLP). The sul1 gene, encoding resistance to SMX at clinically relevant levels, and an element of Class I integrons, was not detected in any community. Changes in microbial community structure and SMX resistance over a short experimental period in previously nonexposed, slow-growing aquifer communities suggest concentrations of antibiotics 2-3 orders of magnitude less than those used in clinical applications may influence ecological function through changes in community composition, and could promote antibiotic resistance through selection of naturally resistant bacteria.

Influence of organic carbon loading, sediment associated metal oxide content and sediment grain size distributions upon Cryptosporidium parvum removal during riverbank filtration operations, Sonoma County, CA.

This study assessed the efficacy for removing Cryptosporidium parvum oocysts of poorly sorted, Fe- and Al-rich, subsurface sediments collected from 0.9 to 4.9 and 1.7-13.9 m below land surface at an operating riverbank filtration (RBF) site (Russian River, Sonoma County, CA). Both formaldehyde-killed oocysts and oocyst-sized (3 microm) microspheres were employed in sediment-packed flow-through and static columns. The degree of surface coverage of metal oxides on sediment grain surfaces correlated strongly with the degrees of oocyst and microsphere removals. In contrast, average grain size (D(50)) was not a good indicator of either microsphere or oocyst removal, suggesting that the primary mechanism of immobilization within these sediments is sorptive filtration rather than physical straining. A low specific UV absorbance (SUVA) for organic matter isolated from the Russian River, suggested that the modest concentration of the SUVA component (0.8 mg L(-1)) of the 2.2 mg L(-1) dissolved organic carbon (DOC) is relatively unreactive. Nevertheless, an amendment of 2.2 mg L(-1) of isolated river DOC to column sediments resulted in up to a 35.7% decrease in sorption of oocysts and (or) oocyst-sized microspheres. Amendments (3.2 microM) of the anionic surfactant, sodium dodecyl benzene sulfonate (SDBS) also caused substantive decreases (up to 31.9 times) in colloid filtration. Although the grain-surface metal oxides were found to have a high colloid-removal capacity, our study suggested that any major changes within the watershed that would result in long-term alterations in either the quantity and (or) the character of the rivers DOC could alter the effectiveness of pathogen removal during RBF operations.

Effects of altered groundwater chemistry upon the pH-dependency and magnitude of bacterial attachment during transport within an organically contaminated sandy aquifer

The effects of a dilute (ionic strength=5x10(-3)M) plume of treated sewage, with elevated levels (3.9 mg/L) of dissolved organic carbon (DOC), upon the pH-dependency and magnitude of bacterial transport through an iron-laden, quartz sand aquifer (Cape Cod, MA) were evaluated using sets of replicate, static minicolumns. Compared with uncontaminated groundwater, the plume chemistry diminished bacterial attachment under mildly acidic (pH 5.0-6.5) in-situ conditions, in spite of the 5-fold increase in ionic strength and substantively enhanced attachment under more alkaline conditions. The effects of the hydrophobic neutral and total fractions of the plume DOC; modest concentrations of fulvic and humic acids (1.5 mg/L); linear alkyl benzene sulfonate (LAS) (25 mg/L); Imbentin (200 microg/L), a model nonionic surfactant; sulfate (28 mg/L); and calcium (20 mg/L) varied sharply in response to relatively small changes in pH, although the plume constituents collectively decreased the pH-dependency of bacterial attachment. LAS and other hydrophobic neutrals (collectively representing only approximately 3% of the plume DOC) had a disproportionately large effect upon bacterial attachment, as did the elevated concentrations of sulfate within the plume. The findings further suggest that the roles of organic plume constituents in transport or bacteria through acidic aquifer sediments can be very different than would be predicted from column studies performed at circumneutral pH and that the inorganic constituents within the plume cannot be ignored.

Differential effects of dissolved organic carbon upon re-entrainment and surface properties of groundwater bacteria and bacteria-sized microspheres during transport through a contaminated, sandy aquifer.

Injection-and-recovery studies involving a contaminated, sandy aquifer (Cape Cod, Massachusetts) were conducted to assess the relative susceptibility for in situ re-entrainment of attached groundwater bacteria (Pseudomonas stuzeri ML2, and uncultured, native bacteria) and carboxylate-modified microspheres (0.2 and 1.0 μm diameters). Different patterns of re-entrainment were evident for the two colloids in response to subsequent injections of groundwater (hydrodynamic perturbation), deionized water (ionic strength alteration), 77 μM linear alkylbenzene sulfonates (LAS, anionic surfactant), and 76 μM Tween 80 (polyoxyethylene sorbitan monooleate, a very hydrophobic nonionic surfactant). An injection of deionized water was more effective in causing detachment of micrsopheres than were either of the surfactants, consistent with the more electrostatic nature of microspheres attachment, their extreme hydrophilicity (hydrophilicity index, HI, of 0.99), and negative charge (zeta potentials, ζ, of -44 to -49 mv). In contrast, Tween 80 was considerably more effective in re-entraining the more-hydrophobic native bacteria. Both the hydrophilicities and zeta potentials of the native bacteria were highly sensitive to and linearly correlated with levels of groundwater dissolved organic carbon (DOC), which varied modestly from 0.6 to 1.3 mg L(-1). The most hydrophilic (0.52 HI) and negatively charged (ζ -38.1 mv) indigenous bacteria were associated with the lowest DOC. FTIR spectra indicated the latter community had the highest average density of surface carboxyl groups. In contrast, differences in groundwater (DOC) had no measurable effect on hydrophilicity of the bacteria-sized microspheres and only a minor effect on their ζ. These findings suggest that microspheres may not be very good surrogates for bacteria in field-scale transport studies and that adaptive (biological) changes in bacterial surface characteristics may need to be considered where there is longer-term exposure to contaminant DOC.

Comparison of transport and attachment behaviors of Cryptosporidium parvum oocysts and oocyst-sized microspheres being advected through three minerologically different granular porous media

In order to gain more information about the fate of Cryptosporidium parvum oocysts in tropical volcanic soils, the transport and attachment behaviors of oocysts and oocyst-sized polystyrene microspheres were studied in the presence of two soils. These soils were chosen because of their differing chemical and physical properties, i.e., an organic-rich (43-46% by mass) volcanic ash-derived soil from the island of Hawaii, and a red, iron (22-29% by mass), aluminum (29-45% by mass), and clay-rich (68-76% by mass) volcanic soil from the island of Oahu. A third agricultural soil, an organic- (13% by mass) and quartz-rich (40% by mass) soil from Illinois, was included for reference. In 10-cm long flow-through columns, oocysts and microspheres advecting through the red volcanic soil were almost completely (98% and 99%) immobilized. The modest breakthrough resulted from preferential flow-path structure inadvertently created by soil-particle aggregation during the re-wetting process. Although a high (99%) removal of oocysts and microsphere within the volcanic ash soil occurred initially, further examination revealed that transport was merely retarded because of highly reversible interactions with grain surfaces. Judging from the slope of the substantive and protracted tail of the breakthrough curve for the 1.8-μm microspheres, almost all (>99%) predictably would be recovered within ∼4000 pore volumes. This suggests that once contaminated, the volcanic ash soil could serve as a reservoir for subsequent contamination of groundwater, at least for pathogens of similar size or smaller. Because of the highly reversible nature of organic colloid immobilization in this soil type, C. parvum could contaminate surface water should overland flow during heavy precipitation events pick up near-surface grains to which they are attached. Surprisingly, oocyst and microsphere attachment to the reference soil from Illinois appeared to be at least as sensitive to changes in pH as was observed for the red, metal-oxide rich soil from Oahu. In contrast, colloidal attachment in the organic-rich, volcanic ash soil was relatively insensitive to changes in pH in spite of the high iron content. Given the fundamental differences in transport behavior of oocyst-sized colloids within the two volcanic soils of similar origin, agricultural practices modified to lessen C. parvum contamination of ground or surface water would necessitate taking the individual soil properties into account.