Bacteriophage tracer experiments in groundwater

Three tracer experiments employing three different bacteriophage were performed at one groundwater site near Beverley, Humberside. In two of the experiments the bacteriophage were injected into the aquifer by a borehole at a distance of 366 m from the pumping borehole. In the other experiment they were injected at a distance of 122 m. Regular samples were taken of water abstracted at the pumping boreholes as well as from the injection boreholes. The objectives were to: (1) investigate the pattern of bacteriophage recovery from the aquifer; (2) calculate the total number of bacteriophage recovered and the rate of their migration; and (3) detect any differences in bacteriophage behaviour which could be directly related to the morphology of the three bacteriophage. In all experiments the pattern of recovery was similar, exhibiting a peak of high numbers reaching the pumping borehole soon after injection. The highest percentage of original inoculum recovered was 1.9%. In the majority of cases, however, recovery was usually one log₁₀ lower than this. The fastest migration rates were very rapid, reaching 2.8 cm/s in one experiment. No variation in percentage recovery or transit time could be directly attributed to morphology of bacteriophage. The most important factor governing the pattern of migration was undoubtedly the hydrogeological conditions.     

Bacteriophage tracer experiments in groundwater

Three tracer experiments employing three different bacteriophage were performed at one groundwater site near Beverley, Humberside. In two of the experiments the bacteriophage were injected into the aquifer by a borehole at a distance of 366 m from the pumping borehole. In the other experiment they were injected at a distance of 122 m. Regular samples were taken of water abstracted at the pumping boreholes as well as from the injection boreholes. The objectives were to: (1) investigate the pattern of bacteriophage recovery from the aquifer; (2) calculate the total number of bacteriophage recovered and the rate of their migration; and (3) detect any differences in bacteriophage behaviour which could be directly related to the morphology of the three bacteriophage. In all experiments the pattern of recovery was similar, exhibiting a peak of high numbers reaching the pumping borehole soon after injection. The highest percentage of original inoculum recovered was 1.9%. In the majority of cases, however, recovery was usually one log10 lower than this. The fastest migration rates were very rapid, reaching 2.8 cm/s in one experiment. No variation in percentage recovery or transit time could be directly attributed to morphology of bacteriophage. The most important factor governing the pattern of migration was undoubtedly the hydrogeological conditions.     

The use of bacteriophage as tracers of aerosols liberated by sludge suction appliances

Three bacteriophage tracers were added to 600-1 containers of water and simulated latrine sludge to provide high titres of tracer in aqueous and semi-aqueous media. After a period of mixing and stabilization, media were removed from the containers with suction hoses coupled to the vacuum pump of one of two sludge suction tankers. Exhaust air from the appliances was sampled by cyclone sampler and assayed for the presence of tracer organisms carried over during the emptying process. In the experiments the appliances were operated at different vacuum pump speeds, drawing both aqueous and semi-aqueous (simulated sludge) media. Air around one tanker was also sampled during the emptying, under pressure, of the vacuum vessel. The degree of aerosolization and expulsion of tracer bacteriophage by the vacuum appliances was consistently low, regardless of medium and pump air flow. In contrast, the proportion of tracer retained within the appliances was very high, exceeding the proportion expelled by a minimum of 8 log10 orders of magnitude and a maximum of greater than 11 log10 orders. The highest total of tracer bacteriophage was recovered during the pressure emptying of the vacuum vessel of one tanker. The results may be used for assessing and comparing potential public health hazards associated with the handling of wastewater sludge by vacuum appliances.     

The use of bacteriophage as tracers of aerosols liberated by sludge suction appliances

Three bacteriophage tracers were added to 600–1 containers of water and simulated latrine sludge to provide high titres of tracer in aqueous and semi-aqueous media. After a period of mixing and stabilization, media were removed from the containers with suction hoses coupled to the vacuum pump of one of two sludge suction tankers. Exhaust air from the appliances was sampled by cyclone sampler and assayed for the presence of tracer organisms carried over during the emptying process. In the experiments the appliances were operated at different vacuum pump speeds, drawing both aqueous and semi-aqueous (simulated sludge) media. Air around one tanker was also sampled during the emptying, under pressure, of the vacuum vessel. The degree of aerosolization and expulsion of tracer bacteriophage by the vacuum appliances was consistently low, regardless of medium and pump air flow. In contrast, the proportion of tracer retained within the appliances was very high, exceeding the proportion expelled by a minimum of 8log₁₀ orders of magnitude and a maximum of greater than 11log₁₀ orders. The highest total of tracer bacteriophage was recovered during the pressure emptying of the vacuum vessel of one tanker. The results may be used for assessing and comparing potential public health hazards associated with the handling of wastewater sludge by vacuum appliances.     

Degradation of 2,4,6-tribromophenol by bacterial cells attached to chalk collected from a contaminated aquifer

AIM: To investigate the factors governing the adhesion and activity of the 2,4,6-tribromophenol (TBP) degrading bacterium Achromobacter piechaudii TBPZ-N61 on chalk from a contaminated aquifer. METHODS AND RESULTS: Adhesion kinetics of TBPZ-N61 to grey and white chalk from a polluted fractured chalk aquifer was tested in a batch system. Both grey and white chalk contain ca 80% CaCO3, while grey chalk contains more organic matter (2.4%) than the white chalk (0.3%) and also contains Dolmite and Clinoptilolite. Adhesion of the bacterial cells to the chalk particles (<0.2 mm) occurred rapidly (96% of the cells within 15 min). Langmuir-fitted adhesion isotherms suggest that cells in the stationary phase, which are more hydrophobic, adhere to both grey and white chalk more efficiently than cells in the logarithmic growth phase. Increasing the pH (from 6.7 to 8.1) caused a significant reduction in cell adhesion to the chalk. Activity of attached cells was evaluated in both batch and column experiments. Logarithmic cells adhering to white and grey chalk were more active in TBP degradation than cells in suspension. In column experiments, significant TBP degradation was retained up to 30 days after a single injection of TBPZ cells. Thereafter, activity was fully recovered by amendment of yeast extract. Chalk surfaces that were incubated in situ in contaminated groundwater for 20 days still allowed the adhesion and activity of TBPZ cells. CONCLUSIONS: Taken together, our results show that bacteria adhere efficiently to specific sites on the chalk surfaces, and that sustained bacterial activity of the attached cells can be achieved by adding a carbon source such as yeast extract which also overcome toxic constituents that may occur in some chalk types. SIGNIFICANCE AND IMPACT OF THE STUDY: Bioremediation of TBP-contaminated chalk aquifers is made possible by the injection of bacterial cultures.     

A Multi-Tracer Push-Pull Diagnostic Test for In Situ Air Sparging Systems

This document describes the development and initial application of a multi-tracer push-pull test designed to provide near real-time point-specific measures of contaminant volatilization and aerobic biodegrada-tion rates during in situ air sparging (IAS) operation. Measured biodegradation and volatilization rates are specific to the tracers used, so the results provide relative measures useful for identifying spatial differences in treatment performance and changes in performance with changes in system operation and design. The diagnostic test involves injecting a solution containing multiple tracer compounds into the target treatment zone through a monitoring well, piezometer, or drive point. The injected solution is initially deoxygenated and can contain: (a) a nondegradable, non-volatile conservative tracer, (b) one or more nondegradable, volatile chemicals, (c) an aerobically biodegradable, nonvolatile compound, and (d) a visible dye. After some predetermined hold time, an excess quantity of groundwater is extracted from the same injection point and the change in the concentrations of the tracer compounds is measured. Volatilization and oxygen utilization rates are then estimated from mass balances on the tracer components. The development of this diagnostic tool was conducted in a controlled physical model study and then initial field tests were conducted at the U.S. Navy Hydrocarbon National Test Site (HNTS) in Port Hueneme, California. Spatial variations in oxygenation and volatilization rates were observed, with oxygenation rates varying from 0 to 51 mg-O₂/L-water/d, and tracer volatilization rates varying from 0 to 47%/d. Acetate and sulfur hexafluoride (SF₆) were used as tracers in the initial testing, and it was discovered that these are not ideal choices due to the potential for anaerobic acetate biodegradation and SF₆ partitioning into trapped gas in the aquifer.     

Composition and diversity of microbial communities recovered from surrogate minerals incubated in an acidic uranium-contaminated aquifer

Our understanding of subsurface microbiology is hindered by the inaccessibility of this environment, particularly when the hydrogeologic medium is contaminated with toxic substances. In this study, surrogate geological media contained in a porous receptacle were incubated in a well within the saturated zone of a pristine region of an aquifer to capture populations from the extant communities. After an 8-week incubation, the media were recovered, and the microbial community that developed on each medium was compared to the community recovered from groundwater and native sediments from the same region of the aquifer, using 16S DNA coding for rRNA (rDNA)-based terminal restriction fragment length polymorphism (T-RFLP). The groundwater and sediment communities were highly distinct from one another, and the communities that developed on the various media were more similar to groundwater communities than to sediment communities. 16S rDNA clone libraries of communities that developed on particles of a specular hematite medium incubated in the same well as the media used for T-RFLP analysis were compared with those obtained from an acidic, uranium-contaminated region of the same aquifer. The hematite-associated community formed in the pristine area was highly diverse at the species level, with 25 distinct phylotypes identified, the majority of which (73%) were affiliated with the beta-Proteobacteria. Similarly, the hematite-associated community formed in the contaminated area was populated in large part by beta-Proteobacteria (62%); however, only 13 distinct phylotypes were apparent. The three numerically dominant clones from the hematite-associated community from the contaminated site were affiliated with metal- and radionuclide-tolerant or acidophilic taxa, consistent with the environmental conditions. Only two populations were common to both sites.     

Using Boreholes as Windows into Groundwater Ecosystems

Groundwater ecosystems remain poorly understood yet may provide ecosystem services, make a unique contribution to biodiversity and contain useful bio-indicators of water quality. Little is known about ecosystem variability, the distribution of invertebrates within aquifers, or how representative boreholes are of aquifers. We addressed these issues using borehole imaging and single borehole dilution tests to identify three potential aquifer habitats (fractures, fissures or conduits) intercepted by two Chalk boreholes at different depths beneath the surface (34 to 98 m). These habitats were characterised by sampling the invertebrates, microbiology and hydrochemistry using a packer system to isolate them. Samples were taken with progressively increasing pumped volume to assess differences between borehole and aquifer communities. The study provides a new conceptual framework to infer the origin of water, invertebrates and microbes sampled from boreholes. It demonstrates that pumping 5 m³ at 0.4–1.8 l/sec was sufficient to entrain invertebrates from five to tens of metres into the aquifer during these packer tests. Invertebrates and bacteria were more abundant in the boreholes than in the aquifer, with associated water chemistry variations indicating that boreholes act as sites of enhanced biogeochemical cycling. There was some variability in invertebrate abundance and bacterial community structure between habitats, indicating ecological heterogeneity within the aquifer. However, invertebrates were captured in all aquifer samples, and bacterial abundance, major ion chemistry and dissolved oxygen remained similar. Therefore the study demonstrates that in the Chalk, ecosystems comprising bacteria and invertebrates extend from around the water table to 70 m below it. Hydrogeological techniques provide excellent scope for tackling outstanding questions in groundwater ecology, provided an appropriate conceptual hydrogeological understanding is applied.     

Transport Behavior of Groundwater Protozoa and Protozoan-Sized Microspheres in Sandy Aquifer Sediments

Transport behaviors of unidentified flagellated protozoa (flagellates) and flagellate-sized carboxylated microspheres in sandy, organically contaminated aquifer sediments were investigated in a small-scale (1 to 4-m travel distance) natural-gradient tracer test on Cape Cod and in flow-through columns packed with sieved (0.5-to 1.0-mm grain size) aquifer sediments. The minute (average in situ cell size, 2 to 3 (mu)m) flagellates, which are relatively abundant in the Cape Cod aquifer, were isolated from core samples, grown in a grass extract medium, labeled with hydroethidine (a vital eukaryotic stain), and coinjected into aquifer sediments along with bromide, a conservative tracer. The 2-(mu)m flagellates appeared to be near the optimal size for transport, judging from flowthrough column experiments involving a polydispersed (0.7 to 6.2 (mu)m in diameter) suspension of carboxylated microspheres. However, immobilization within the aquifer sediments accounted for a log unit reduction over the first meter of travel compared with a log unit reduction over the first 10 m of travel for indigenous, free-living groundwater bacteria in earlier tests. High rates of flagellate immobilization in the presence of aquifer sediments also was observed in the laboratory. However, immobilization rates for the laboratory-grown flagellates (initially 4 to 5 (mu)m) injected into the aquifer were not constant and decreased noticeably with increasing time and distance of travel. The decrease in propensity for grain surfaces was accompanied by a decrease in cell size, as the flagellates presumably readapted to aquifer conditions. Retardation and apparent dispersion were generally at least twofold greater than those observed earlier for indigenous groundwater bacteria but were much closer to those observed for highly surface active carboxylated latex microspheres. Field and laboratory results suggest that 2-(mu)m carboxylated microspheres may be useful as analogs in investigating several abiotic aspects of flagellate transport behavior in groundwater.     

Impact of groundwater abstraction on a Banksia woodland, Swan Coastal Plain, Western Australia

Summary The Gnangara Groundwater Mound, centred 38 km north of Perth, Western Australia, is a large, shallow unconfined aquifer that is currently under abstraction as part of the public metropolitan water supply. To investigate the impact of lowering groundwater levels on a Banksia woodland on the Mound, vegetation monitoring near a groundwater abstraction bore (known as P50) began 1 year before becoming operational. In February 1991, 2 years after abstraction commenced, extensive death of the Banksia overstorey was observed within close proximity of the bore, following a short period of high summer temperatures. The site was subsequently revisited and the understorey floristic composition, abundance and vigour of overstorey species resurveyed, and compared with data collected from a site under long‐term monitoring and not currently influenced by abstraction. A lowering of groundwater level by 2.2 m at P50 between the summers of 1990 and 1991, resulting from the cumulative effects of abstraction and below average annual rainfall (low groundwater recharge), coincided with a loss of between 20 and 80% of adults of overstorey species and up to 64% of adults of understorey species within 200 m of the bore. Over a similar time period no significant decreases in the abundance of overstorey or understorey species were recorded in the monitored site not influenced by groundwater abstraction. Of the overstorey species, Holly‐leaf Banksia (Banksia ilicifolia) displayed the greatest susceptibility and lowest net recovery following the abstraction event at P50. The negative impact of groundwater drawdown on Holly‐leaf Banksia populations makes this overstorey species an important indicator of decreasing groundwater levels on the Gnangara Groundwater Mound. Water stress may have been the primary cause of vegetation death in close proximity to the P50 bore, although this would have been exacerbated by extreme summer temperatures (> 45°C) recorded during February 1991. The P50 scenario represents a localized response to an acute drawdown event, in association with other environmental factors, and provides invaluable information on the assessment of groundwater abstraction and poor groundwater recharge events on a Banksia woodland community. However, there are limitations in using the community response at P50 to manage the impact of drawdown events on other plant communities occurring on sandy, shallow aquifers.     

In situ measurement of methane oxidation in groundwater by using natural-gradient tracer tests

Methane oxidation was measured in an unconfined sand and gravel aquifer (Cape Cod, Mass.) by using in situ natural-gradient tracer tests at both a pristine, oxygenated site and an anoxic, sewage-contaminated site. The tracer sites were equipped with multilevel sampling devices to create target grids of sampling points; the injectate was prepared with groundwater from the tracer site to maintain the same geochemical conditions. Methane oxidation was calculated from breakthrough curves of methane relative to halide and inert gas (hexafluroethane) tracers and was confirmed by the appearance of 13C-enriched carbon dioxide in experiments in which 13C-enriched methane was used as the tracer. A Vmax for methane oxidation could be calculated when the methane concentration was sufficiently high to result in zero-order kinetics throughout the entire transport interval. Methane breakthrough curves could be simulated by modifying a one-dimensional adevection-dispersion transport model to include a Michaelis-Menten-based consumption term for methane oxidation. The Km values for methane oxidation that gave the best match for the breakthrough curve peaks were 6.0 and 9.0 microM for the uncontaminated and contaminated sites, respectively. Natural-gradient tracer tests are a promising approach for assessing microbial processes and for testing in situ bioremediation potential in groundwater systems.     

In situ measurement of methane oxidation in groundwater by using natural-gradient tracer tests.

Methane oxidation was measured in an unconfined sand and gravel aquifer (Cape Cod, Mass.) by using in situ natural-gradient tracer tests at both a pristine, oxygenated site and an anoxic, sewage-contaminated site. The tracer sites were equipped with multilevel sampling devices to create target grids of sampling points; the injectate was prepared with groundwater from the tracer site to maintain the same geochemical conditions. Methane oxidation was calculated from breakthrough curves of methane relative to halide and inert gas (hexafluroethane) tracers and was confirmed by the appearance of 13C-enriched carbon dioxide in experiments in which 13C-enriched methane was used as the tracer. A Vmax for methane oxidation could be calculated when the methane concentration was sufficiently high to result in zero-order kinetics throughout the entire transport interval. Methane breakthrough curves could be simulated by modifying a one-dimensional adevection-dispersion transport model to include a Michaelis-Menten-based consumption term for methane oxidation. The Km values for methane oxidation that gave the best match for the breakthrough curve peaks were 6.0 and 9.0 microM for the uncontaminated and contaminated sites, respectively. Natural-gradient tracer tests are a promising approach for assessing microbial processes and for testing in situ bioremediation potential in groundwater systems.     

Composition and Diversity of Microbial Communities Recovered from Surrogate Minerals Incubated in an Acidic Uranium-Contaminated Aquifer

Our understanding of subsurface microbiology is hindered by the inaccessibility of this environment, particularly when the hydrogeologic medium is contaminated with toxic substances. In this study, surrogate geological media contained in a porous receptacle were incubated in a well within the saturated zone of a pristine region of an aquifer to capture populations from the extant communities. After an 8-week incubation, the media were recovered, and the microbial community that developed on each medium was compared to the community recovered from groundwater and native sediments from the same region of the aquifer, using 16S DNA coding for rRNA (rDNA)-based terminal restriction fragment length polymorphism (T-RFLP). The groundwater and sediment communities were highly distinct from one another, and the communities that developed on the various media were more similar to groundwater communities than to sediment communities. 16S rDNA clone libraries of communities that developed on particles of a specular hematite medium incubated in the same well as the media used for T-RFLP analysis were compared with those obtained from an acidic, uranium-contaminated region of the same aquifer. The hematite-associated community formed in the pristine area was highly diverse at the species level, with 25 distinct phylotypes identified, the majority of which (73%) were affiliated with the β-Proteobacteria. Similarly, the hematite-associated community formed in the contaminated area was populated in large part by β-Proteobacteria (62%); however, only 13 distinct phylotypes were apparent. The three numerically dominant clones from the hematite-associated community from the contaminated site were affiliated with metal- and radionuclide-tolerant or acidophilic taxa, consistent with the environmental conditions. Only two populations were common to both sites.     

Related assemblages of sulphate-reducing bacteria associated with ultradeep gold mines of South Africa and deep basalt aquifers of Washington State

We characterized the diversity of sulphate-reducing bacteria (SRB) associated with South African gold mine boreholes and deep aquifer systems in Washington State, USA. Sterile cartridges filled with crushed country rock were installed on two hydrologically isolated and chemically distinct sites at depths of 3.2 and 2.7 km below the land surface (kmbls) to allow development of biofilms. Enrichments of sulphate-reducing chemolithotrophic (H2) and organotrophic (lactate) bacteria were established from each site under both meso- and thermophilic conditions. Dissimilatory sulphite reductase (Dsr) and 16S ribosomal RNA (rRNA) genes amplified from DNA extracted from the cartridges were most closely related to the Gram-positive species Desulfotomaculum thermosapovorans and Desulfotomaculum geothermicum, or affiliated with a novel deeply branching clade. The dsr sequences recovered from the Washington State deep aquifer systems affiliated closely with the South African sequences, suggesting that Gram-positive sulphate-reducing bacteria are widely distributed in the deep subsurface.     

Denitrification in a Sand and Gravel Aquifer

Denitrification was assayed by the acetylene blockage technique in slurried core material obtained from a freshwater sand and gravel aquifer. The aquifer, which has been contaminated with treated sewage for more than 50 years, had a contaminant plume greater than 3.5-km long. Near the contaminant source, groundwater nitrate concentrations were greater than 1 mM, whereas 0.25 km downgradient the central portion of the contaminant plume was anoxic and contained no detectable nitrate. Samples were obtained along the longitudinal axis of the plume (0 to 0.25 km) at several depths from four sites. Denitrification was evident at in situ nitrate concentrations at all sites tested; rates ranged from 2.3 to 260 pmol of N₂O produced (g of wet sediment)⁻¹ h⁻¹. Rates were highest nearest the contaminant source and decreased with increasing distance downgradient. Denitrification was the predominant nitrate-reducing activity; no evidence was found for nitrate reduction to ammonium at any site. Denitrifying activity was carbon limited and not nitrate limited, except when the ambient nitrate level was less than the detection limit, in which case, even when amended with high concentrations of glucose and nitrate, the capacity to denitrify on a short-term basis was lacking. These results demonstrate that denitrification can occur in groundwater systems and, thereby, serve as a mechanism for nitrate removal from groundwater.     

Virus and Bacteria Removal from Wastewater by Rapid Infiltration Through Soil

A rapid infiltration land wastewater application site, composed of unconsolidated silty sand and gravel, which has been in continuous operation for over 30 years was examined for the accumulation and/or migration of a tracer virus (coliphage f2), indigenous enteroviruses, and enteric indicator bacteria in the soils and underlying groundwater. Tracer f2 penetrated into groundwater together with the front of percolating primary effluent and was not observed to concentrate on the upper soil layers. The tracer virus concentration in a 60-foot (about 18.3-m)-deep observation well directly beneath the wastewater application area began to increase within 48 h after application to the soil. The tracer level in this well stabilized after 72 h at a level of approximately 47% of the average applied concentration. Indigenous enteroviruses and tracer f2 were sporadically detected in the groundwater at horizontal distances of 600 feet (about 183 m) from the application zone. Laboratory soil adsorption studies confirmed the poor virus adsorption observed at the site. This was especially true on surface soils when contained in wastewater. Enteric indicator bacteria were readily concentrated on the soil surface by filtration on the soil surface mat. However, during tracer f2 virus tests, comparison studies with fecal Streptococcus revealed that bacteria capable of penetrating the surface were able to migrate into the groundwater. They were detected at the same locations as tracer and enteric viruses.     

Denitrification in Aquifer Soil and Nearshore Marine Sediments Influenced by Groundwater Nitrate

We estimated rates of denitrification at various depths in sediments known to be affected by submarine discharge of groundwater, and also in the parent aquifer. Surface denitrification was only measured in the autumn; at 40-cm depth, where groundwater-imported nitrate has been measured, denitrification occurred consistently throughout the year, at rates from 0.14 to 2.8 ng-atom of N g⁻¹ day⁻¹. Denitrification consistently occurred below the zone of sulfate reduction and was sometimes comparable to it in magnitude. Denitrification occurred deep (14 to 40 cm) in the sediments along 30 km of shoreline, with highest rates occurring where groundwater input was greatest. Denitrification rates decreased with distance offshore, as does groundwater influx. Added glucose greatly stimulated denitrification at depth, but added nitrate did not. High rates of denitrification were measured in the aquifer (17 ng-atom of N g⁻¹ day⁻¹), and added nitrate did stimulate denitrification there. The denitrification measured was enough to remove 46% of the nitrate decrease observed between 40- and 14-cm depth in the sediment.     

On the possibility for generic modeling of submarine groundwater discharge

We simulate large-scale dynamics of submarine groundwater discharge (SGD) in three different coastal aquifers on the Mediterranean Sea. We subject these aquifers to a wide range of different groundwater management conditions, leading to widely different net groundwater drainage from land to sea. The resulting SGD at steady-state is quantifiable and predictable by simple linearity in the net land-determined groundwater drainage, defined as total fresh water drainage minus groundwater extraction in the coastal aquifer system. This linearity appears to be general and independent of site-specific, variable and complex details of hydrogeology, aquifer hydraulics, streamlines and salinity transition zones in different coastal systems. Also independently of site-specifics, low SGD implies high seawater content due to seawater intruding into the aquifer and mixing with fresh groundwater within a wide salinity transition zone in the aquifer. Increasing SGD implies decreasing seawater content, decreased mixing between seawater and fresh groundwater and narrowing of the salinity transition zone of brackish groundwater in the aquifer.     

The Distribution of Microbial Communities in Anaerobic and Aerobic Zones of a Shallow Coastal Plain Aquifer

Abstract Randomly amplified polymorphic DNA (RAPD) fingerprinting was used to determine the genetic similarity of whole-community DNA extracts from unattached microorganisms in several groundwater wells. The study site was a shallow coastal plain aquifer on the Eastern Shore of Virginia that contains distinct regions of anaerobic and aerobic groundwater. Several wells in each region were sampled, and principal component and cluster analyses showed a clear separation of the microbial communities from the two chemical zones of the aquifer. Within these zones, there was no relationship between the genetic relatedness of a pair of communities and their spatial separation. Two additional sets of samples were taken at later times, and the same clear separation between communities in the different zones of the aquifer was observed. The specific relationships between wells within each zone changed over time, however, and the magnitude and direction of these changes corresponded to concurrent changes in the groundwater chemistry at each well. Together, these results suggest that local variation in groundwater chemistry can support genetically distinct microbial communities, and that the composition of the microbial communities can follow seasonal fluctuations in groundwater chemistry. Received: 25 May 1999; Accepted: 4 August 1999; Online Publication: 9 December 1999     

Measurement of lung water in dog lobes using inhaled C15O2 and injected H215O

Indicator-dilution analysis was used in a recirculation-free isolated dog lobe preparation to compare an inhaled water tracer (C15O2) and an injected water tracer (H215O) with direct weighing as a measure of total lung water. Residue detection (counting over the lung) was compared with outflow detection (counting over the venous effluent). With outflow detection, inhaled C15O2 measured 74% and injected H215O 90% of the gravimetric lung water. In hemodynamic edema, the increase in lung water measured by residue detection of both tracers correlated well with increases in weight (r = 0.92, slope = 1.03). However, outflow detection of both tracers underestimated the lung water increase by 53% in edema (r = 0.88, slope = 0.47). Thus, in edema, equilibration of both tracers within the lung water volume is rapid, but clearance from the lung is delayed because slowly clearing water pools develop. The errors caused by inhomogeneity of perfusion distribution were investigated after pulmonary arterial injection of 34-, 50-, and 175-micrometers spheres. For the same lung weight, C15O2 transit was delayed and H215O transit accelerated greatly by the 175-micrometers spheres and slightly by the 50-micrometers spheres.