A microbial community analysis using 16S rRNA gene sequencing was performed on borehole water and a granite rock core from Henderson Mine, a >1,000-meter-deep molybdenum mine near Empire, CO. Chemical analysis of borehole water at two separate depths (1,044 m and 1,004 m below the mine entrance) suggests that a sharp chemical gradient exists, likely from the mixing of two distinct subsurface fluids, one metal rich and one relatively dilute; this has created unique niches for microorganisms. The microbial community analyzed from filtered, oxic borehole water indicated an abundance of sequences from iron-oxidizing bacteria (Gallionella spp.) and was compared to the community from the same borehole after 2 weeks of being plugged with an expandable packer. Statistical analyses with UniFrac revealed a significant shift in community structure following the addition of the packer. Phospholipid fatty acid (PLFA) analysis suggested that Nitrosomonadales dominated the oxic borehole, while PLFAs indicative of anaerobic bacteria were most abundant in the samples from the plugged borehole. Microbial sequences were represented primarily by Firmicutes, Proteobacteria, and a lineage of sequences which did not group with any identified bacterial division; phylogenetic analyses confirmed the presence of a novel candidate division. This "Henderson candidate division" dominated the clone libraries from the dilute anoxic fluids. Sequences obtained from the granitic rock core (1,740 m below the surface) were represented by the divisions Proteobacteria (primarily the family Ralstoniaceae) and Firmicutes. Sequences grouping within Ralstoniaceae were also found in the clone libraries from metal-rich fluids yet were absent in more dilute fluids. Lineage-specific comparisons, combined with phylogenetic statistical analyses, show that geochemical variance has an important effect on microbial community structure in deep, subsurface systems.

Simplified injection, sampling, and analytical procedures using dissolved gases as groundwater tracers are presented for use in saturated conditions at both the laboratory and field scales. The injection of gases into the groundwater is accomplished by allowing the gas to diffuse through semipermeable tubing, minimizing the formation of bubbles that could modify the hydraulic properties around the well. We have simplified the collection of dissolved gases by developing a passive in situ headspace sampler the employs a semipermeable membrane and copper tubing equipped with a schrader valve. The headspace within the sampler equilibrates with the dissolved gases in the groundwater in around 24 hours, and no groundwater is collected, which is of great advantage for use in contaminated sites. The design parameters and the time to equilibrium of the headspace sampler can be adjusted for investigation requirements using the analytical equation presented. The analysis of the gases for tracer content is performed by using a common gas Chromatograph fitted with a thermal conductivity detector. Examples of the use of these methods at both the laboratory and field scales are presented.

A significant limitation in defining remediation needs at contaminated sites often results from an insufficient understanding of the transport processes that control contaminant migration. The objectives of this research were to help resolve this dilemma by providing an improved understanding of contaminant transport processes in highly structured, heterogeneous subsurface environments that are complicated by fracture flow and matrix diffusion. Our approach involved a unique long‐term, steady state natural gradient injection of multiple tracers with different diffusion coefficients (Br, He, Ne) into a fracture zone of a contaminated shale bedrock. The spatial and temporal distribution of the tracers was monitored for 550 days using an array of groundwater sampling wells instrumented within a fast flowing fracture regime and a slow flowing matrix regime. The tracers were transported preferentially along strike‐parallel fractures, with a significant portion of the tracer plumes migrating slowly into the bedrock matrix. Movement into the matrix was controlled by concentration gradients established between preferential flow paths and the adjacent rock matrix. Observed differences in tracer mobility into the matrix were found to be a function of their free‐water molecular diffusion coefficients. The multiple tracer technique confirmed that matrix diffusion was a significant process that contributed to the overall physical nonequilibrium that controlled contaminant transport in the shale bedrock. The experimental observations were consistent with numerical simulations of the multitracer breakthrough curves using a simple fracture flow model. The simulated results also demonstrated the significance of contaminant diffusion into the bedrock matrix. The multiple tracer technique and ability to monitor the fracture and matrix regimes provided the necessary experimental constraints for the accurate numerical quantification of the diffusive mass transfer process. The experimental and numerical results of the tracer study were also consistent with indigenous contaminant discharge concentrations within the fracture and matrix regimes of the field site. These findings suggest that the secondary source contribution of the bedrock matrix to the total off‐site transport of contaminants is relatively large and potentially long‐lived.

A field‐scale tracer experiment carried out under natural gradient ground water flow conditions showed that colloids can be highly mobile in a fractured and highly weathered shale saprolite. Four colloidal tracers (0.100 μm fluorescent latex microspheres, bacteriophage strains PRD‐1 and MS‐2, and INA, a dead strain of Pseudemonas syringae), were introduced to a 6.4 m deep well, and concentrations of the tracers were monitored in the source well and in downgradient monitoring wells at distances of 2 to 35 m. All of the colloidal tracers were detected to distances of at least 13.5 m and two of the tracers (microspheres and INA) were detected in all of the downgradient wells. In most wells the colloidal tracers appeared as a “pulse”, with rapid first arrival (corresponding to 5 to 200 m/d transport velocity), one to six days of high concentrations, and then a rapid decline to below the detection limit. The colloids were transported at velocities of up to 500 times faster than solute tracers (He, Ne, and rhodamine‐WT) from previous tests at the site. This is believed to be largely due to greater diffusion of the solutes into the relatively immobile pore water of the fine‐grained matrix between fractures. Peak colloid tracer concentrations in the monitoring wells varied substantially, with the microspheres exhibiting the highest relative concentrations and hence the least retention. Rates of concentration decline with distance also varied, indicating that retention is not a uniform process in this heterogeneous material. Two of the tracers, INA and PRD‐1, reappeared in several monitoring wells one to five months after the initial pulse had passed, and the reappearance generally corresponds with increased seasonal precipitation. This is consistent with subsequent laboratory experiments that showed that colloid retention in these materials is sensitive to factors such as flow rate and ionic strength, both of which are expected to vary with the amount of precipitation.

The Ridge and Valley Province of eastern Tennessee is characterized by (1) substantial topographic relief, (2) folded and highly fractured rocks of various lithologies that have low primary permeability and porosity, and (3) a shallow residuum of medium permeability and high total porosity. Conceptual models of shallow groundwater flow and solute transport in this system have been developed but are difficult to evaluate using physical characterization or short‐term tracer methods due to extreme spatial variability in hydraulic properties. In this paper we describe how chlorofluorocarbon 12, 3H, and 3He were used to infer groundwater flow and solute transport in saprolite and fractured rock near Oak Ridge, Tennessee. In the shallow residuum, fracture spacings are <0.05 m, suggesting that concentrations of these tracers in fractures and in the matrix have time to diffusionally equilibrate. The relatively smooth nature of tracer concentrations with depth in the residuum is consistent with this model and quantitatively suggests recharge fluxes of 0.2 to 0.4 m yr−1. In contrast, groundwater flow within the unweathered rock appears to be controlled by fractures with spacings of the order of 2 to 5 m, and diffusional equilibration of fractures and matrix has not occurred. For this reason, vertical fluid fluxes in the unweathered rock cannot be estimated from the tracer data.