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Groundwater-quality and select quality-control data from the National Water-Quality Assessment Project, January 2017 through December 2019

Groundwater-quality environmental data were collected from 983 wells as part of the National Water-Quality Assessment Project of the U.S. Geological Survey National Water Quality Program and are included in this report. The data were collected from six types of well networks: principal aquifer study networks, which are used to assess the quality of groundwater used for public water supply; land-use study networks, which are used to assess land-use effects on shallow groundwater quality; major aquifer study networks, which are used to assess the quality of groundwater used for domestic supply; enhanced trends networks, which are used to evaluate the time scales during which groundwater quality changes; vertical flow-path study networks, which are used to evaluate changes in groundwater quality from shallow to deeper depths; and modeling support studies, which are used to provide data to support groundwater modeling. Groundwater samples were analyzed for many water-quality indicators and constituents, including major ions, nutrients, trace elements, volatile organic compounds, pesticides, radionuclides, microbiological indicators, and some constituents of special interest (arsenic speciation, hexavalent chromium [chromium (VI)], and perchlorate). These groundwater-quality data, along with data from quality-control samples, are tabulated in this report and in an associated data release. Data for microbiological indicators for samples collected in 2016 are included in the companion data release.

Data Series

Hydrogeology and water quality of the Leetown area, West Virginia

The U.S. Geological Survey’s Leetown Science Center and the co-located U.S. Department of Agriculture’s National Center for Cool and Cold Water Aquaculture both depend on large volumes of cold clean ground water to support research operations at their facilities. Currently, ground-water demands are provided by three springs and two standby production wells used to augment supplies during periods of low spring flow. Future expansion of research operations at the Leetown Science Center is dependent on assessing the availability and quality of water to the facilities and in locating prospective sites for additional wells to augment existing water supplies. The hydrogeology of the Leetown area, West Virginia, is a structurally complex karst aquifer. Although the aquifer is a karst system, it is not typical of most highly cavernous karst systems, but is dominated by broad areas of fractured rock drained by a relatively small number of solution conduits. Characterization of the aquifer by use of fluorometric tracer tests, a common approach in most karst terranes, therefore only partly defines the hydrogeologic setting of the area. In order to fully assess the hydrogeology and water quality in the vicinity of Leetown, a multi-disciplinary approach that included both fractured rock and karst research components was needed. The U.S. Geological Survey developed this multi-disciplinary research effort to include geologic, hydrologic, geophysical, geographic, water-quality, and microbiological investigations in order to fully characterize the hydrogeology and water quality of the Leetown area, West Virginia. Detailed geologic and karst mapping provided the framework on which hydrologic investigations were based. Fracture trace and lineament analysis helped locate potential water-bearing fractures and guided installation of monitoring wells. Monitoring wells were drilled for borehole geophysical surveys, water-quality sampling, water-level measurements, and aquifer tests to characterize the quality of water and the hydraulic properties of the aquifer. Surface geophysical surveys provided a 3-dimensional view of bedrock resistivity in order to assess geologic and lithologic controls on ground-water flow. Borehole geophysical surveys were conducted in monitoring wells to assess the storage and movement of water in subsurface fractures. Numerous single-well, multi-well, and straddle packer aquifer tests and step-drawdown tests were conducted to define the hydraulic properties of the aquifer and to assess the role of bedrock fractures and solution conduits in the flow of ground water. Water samples collected from wells and springs were analyzed to assess the current quality of ground water and provide a baseline for future assessment. Microbiological sampling of wells for indicator bacteria and human and animal DNA provided an analysis of agricultural and suburban development impacts on ground-water quality. Light detection and ranging (LiDAR) data were analyzed to develop digital elevation models (DEMs) for assessing sinkhole distribution, to provide elevation data for development of a ground-water flow model, and to assess the distribution of major fractures and faults in the Leetown area. The flow of ground water in the study area is controlled by lithology and geologic structure. Bedrock, especially low permeability units such as the shale Martinsburg Formation and the Conococheague Limestone, act as barriers to water flowing down gradient and across bedding. This retardation of cross-strike flow is especially pronounced in the Leetown area, where bedding typically dips at steep angles. Highly permeable fault and fracture zones that disrupt the rocks in cross-strike directions provide avenues through which ground water can flow laterally across or through strata of low primary permeability. Significant strike parallel thrust faults and cross-strike faults typically coincide with larger solution conduits and act as drains for the more pervasive network of interconnected diffuse fractures. Results of borehole geophysical surveys indicate that although numerous fractures may intersect a borehole, only one or two of the fractures typically transmit most of the water to a well. The diffuse-flow dominated network of fractures that provides the majority of storage occupies only a small proportion of the total aquifer volume but constitutes the majority of porosity within the aquifer. Solution conduits, while occupying a relatively small volume of the overall aquifer, are especially important because they serve as primary drains for the ground-water flow system. Surface resistivity maps and cross-sectionsshow anomalous areas of low resistivities coincident with the prevailing geologic strike at N. 20º E., with major cross-strike faults, and with major springs in the region. Transmissivity derived from straddle packer tests was highly variable, and ranged over three orders of magnitude (1.8 x 10 -6 to 5.9 x 10 -3 ft 2 /d) in diffuse-flow fractures. A similar large variability in transmissivity was documented by single- and multi-well aquifer tests conducted in conduit-flow dominated portions of the aquifer (2.0 x 10 3 to 1.4 x 10 4 ft 2 /d) in lowland areas immediately adjacent to the Leetown Science Center. A stream-gaging station installed on Hopewell Run near the point where the stream exits the Leetown watershed indicates average daily streamflow for the Hopewell Run of approximately 11.2 ft 3 /s, and ranged from a minimum of 1.80 ft 3 /s on September 28, 2005, to a maximum of 73.0 ft 3 /s on December 11, 2003. Base-flow (ground-water) discharge surveys identified numerous small seeps adjacent to streams in the area. Hydrographs of the stage of Balch Spring show rapid response to individual storms. Strong correlation of the flow of Hopewell Run and Balch Spring indicates the nearby losing stream reach is partly responsible for higher fluctuations in the stage of Balch Spring. A water budget for the study period (2003-2005), based on measured precipitation and hydrograph analyses, is expressed as Precipitation (38.60 in/yr) = Surface Runoff (1.36 in/yr) + Ground-Water Discharge (17.73 in/yr) + Evapotranspiration (24.23 in/yr) – Change in storage (4.72 in/yr). Flow of ground water through the epikarst, a shallow zone of intensely weathered rock and regolith, can be rapid (on the order of days or weeks) as flow is concentrated in solution conduits. Flow within the intermediate and deeper zones is typically much slower. Eight dye-tracer tests conducted in the Leetown area found ground-water flow patterns to be divergent, with velocities ranging from about 12.5 to 610 ft/day and a median velocity of 50 ft/day. Estimates of ground-water age in carbonate rocks in the region are on the order of 15 years in the shallower portions of the aquifer to 50 years or older for deeper portions of the aquifer. Shallow springs can have a significant component of fairly young water (< 5 years in age). Ground-water samples collected from 16 sites (12 wells and 4 springs) in the Leetown area were analyzed for more than 340 constituents. Only turbidity, indicator bacteria, and radon were typically present in concentrations exceeding U.S. Environmental Protection Agency (USEPA) drinking-water or aquatic life standards.

West Virginia

Microbial and dissolved organic carbon characterization of stormflow in the Santa Ana River at Imperial Highway, Southern California, 1999-2002

The Santa Ana River drains about 2,670 square miles of densely populated coastal southern California, near Los Angeles. Almost all the flow in the river, more than 200,000 acre-feet annually, is diverted to ponds where it infiltrates and recharges underlying aquifers pumped to supply water for more than 2 million people. Base flow in the river is almost entirely treated municipal wastewater discharged from upstream treatment plants and, in the past, stormflow was considered a source of high-quality water suitable for use as a source of ground-water recharge that would dilute poorer quality water recharged during base flow. Stormflow in the Santa Ana River at the Imperial Highway diversion contains total coliform bacteria concentrations as high as 3,400,000 colonies per 100 mL (milliliters). Fecal indicator bacteria concentrations, including fecal coliforms, Escherichia coli, and enterococci, were as high as 310,000, 84,000, and 102,000 colonies per 100 mL, respectively. Although concentrations were high owing to urban runoff during the first stormflow of the rainy season, the highest concentrations occurred during the recessional flows of the first stormflow of the rainy season after streamflow returned to pre-storm conditions. Molecular indicators of microbiological organisms in stormflow, including phospholipid fatty acid (PLFA) and genetic data, show that the diversity of the total microbial population decreases during stormflow while fecal indicator bacteria concentrations increase. This suggests that the source of the bacteria must be poorly diverse and dominated by only a few types of bacteria. Although direct runoff of fecal indicator bacteria from urban areas occurs, this process cannot explain the very high concentrations of fecal indicator bacteria in runoff from upstream parts of the basin characterized by urban, agricultural (including more than 300,000 head of dairy cattle), and other land uses. Although other explanations are possible, fecal indicator bacteria concentrations and molecular microbiological data indicate accumulation and extended survival of bacteria in streambed sediments, and subsequent mobilization of those sediments and associated bacteria during stormflow. Both PLFA and genetic data indicate that water from dairy-waste storage ponds was not present during sampled stormflows. This is consistent with the relatively dry conditions and the absence of large stormflows during the study. Dissolved organic carbon (DOC) concentrations in stormflow ranged from 3 to 15.3 mg/L. In general, concentrations increased during stormflow and were distributed across the stormflow hydrograph in a manner similar to that of fecal indicator bacteria. DOC concentrations typically remained high for several days after flow returned to pre-storm conditions. Ultraviolet absorbance, excitation emission spectroscopy, and sequential fractionation of DOC using XAD-8 and XAD-4 resins showed that the composition of DOC changed rapidly during stormflow. Hydrophobic and hydrophilic acids were the largest fraction of DOC composing between 27 and 45 percent and between 24 and 37 percent of the DOC, respectively. The fraction of DOC composed of hydrophobic acids decreased due to urban runoff and increased during the recession of the first stormflow of the rainy season; the hydrophilic-acid fraction generally decreased throughout the stormflow hydrograph; the transhydrophilic-acid fraction did not vary greatly during stormflow; and the hydrophobic-neutral fraction increased from low values in base flow to almost 30 percent of the DOC after more soluble and more mobile hydrophobic and hydrophilic acids were washed from urban areas. Comparison of ultraviolet absorbance data with data collected during previous studies shows that the optical properties and, presumably, the composition of the DOC were different in this study than DOC collected during wetter periods. Samples of shallow ground water collec

California

The effects of artificial recharge on groundwater levels and water quality in the west hydrogeologic unit of the Warren subbasin, San Bernardino County, California

Between the late 1940s and 1994, groundwater levels in the Warren subbasin, California, declined by as much as 300 feet because pumping exceeded sparse natural recharge. In response, the local water district, Hi-Desert Water District, implemented an artificial-recharge program in early 1995 using imported water from the California State Water Project. Subsequently, the water table rose by as much as 250 feet; however, a study done by the U.S. Geological Survey found that the rising water table entrained high-nitrate septic effluent, which caused nitrate (as nitrogen) concentrations in some wells to increase to more than the U.S. Environmental Protection Agency maximum contaminant level of 10 milligrams per liter.. A new artificial-recharge site (site 3) was constructed in 2006 and this study, which started in 2004, was done to address concerns about the possible migration of nitrates in the unsaturated zone. The objectives of this study were to: (1) characterize the hydraulic, chemical, and microbiological properties of the unsaturated zone; (2) monitor changes in water levels and water quality in response to the artificial-recharge program at site 3; (3) determine if nitrates from septic effluent infiltrated through the unsaturated zone to the water table; (4) determine the potential for nitrates within the unsaturated zone to mobilize and contaminate the groundwater as the water table rises in response to artificial recharge; and (5) determine the presence and amount of dissolved organic carbon because of its potential to react with disinfection byproducts during the treatment of water for public use. Two monitoring sites were installed and instrumented with heat-dissipation probes, advanced tensiometers, suction-cup lysimeters, and wells so that the arrival and effects of recharging water from the State Water Project through the 250 to 425 foot-thick unsaturated zone and groundwater system could be closely observed. Monitoring site YVUZ-1 was located between two recharge ponds in the middle of site 3, and YVUZ-2 was located approximately 1,200 feet down-gradient and to the southeast in an area where septic systems have been in use since about 1960. Site YVUZ-3 only went to a depth of 42 feet and was used to sample the upper part of the unsaturated zone near a golf course. Prior to the start of artificial recharge at site 3, nitrate concentrations reported as nitrogen from the soil leachate below YVUZ-1 did not exceed 1.58 milligrams per kilogram. Nitrate-reducing bacteria concentrations of 4,300 most probable number were found at about 220 feet below land surface and at the top of the water table at YVUZ-1. Nitrate concentrations at YVUZ-2 reached a maximum concentration of about 25 milligrams per kilogram between about 100 and 121 feet below land surface; concentrations of nitrate-reducing or denitrifying bacteria were as high as 21,000 most probable number at 36 feet below land surface but did not exceed 40 most probable number below about 150 feet below land surface. Between June 2006 and September 2009, more than 9,800 acre feet of water from the State Water Project was released to site 3 ponds. The infiltration of the recharge water was predominantly vertical with limited lateral spreading to a depth of about 200 feet below land surface at YVUZ-1. Lateral spreading of the recharge water with depth was caused by geologic heterogeneities within the unsaturated zone, and resulted in varied arrival times of the recharge water to the instruments and slower rates of vertical movement with depth. No abrupt changes in soil moisture were observed at YVUZ-2, indicating that the recharge water had not reached that site by September 2009. Water levels from the monitoring wells at both sites and from five production wells nearby showed that the water table rose at a mean rate of about 0.08 feet per day between June 2006 and January 2009. The arrival of the water from the State Water Project caused relatively rapid changes in the stable-isotopic ratios from the lysimeters at YVUZ-1. The estimated average rate of infiltration of the recharge water through the unsaturated zone ranged from 3.7 to 25 feet per day. The recharge water arrived at the monitoring well below the recharge ponds between August 2007 and March 2008; the rate of vertical movement to the monitoring well was between 0.6 and 0.9 feet per day. By September 2008, a production well located 375 feet west of site 3 was producing almost 100 percent infiltrated recharge water. By contrast, the stable-isotope data from the lysimeters at YVUZ-2 showed that the recharge water had not reached this site by September 2009, but that septic effluent in the unsaturated zone likely had mixed with the native pore water to at least 154 feet below land surface. Assuming vertical infiltration, the minimum rate of infiltration of septic effluent since 1960 was about 3 feet per year. The isotopic data from the lysimeters at YVUZ-3 indicated two different sources of water to the upper 43 feet–irrigation-return flow and precipitation. Nitrate concentrations of the water from the State Water Project did not exceed 1 milligram per liter. Prior to artificial recharge, nitrate concentrations of the pore water at YVUZ-1 ranged between 6 to 18.2 milligrams per liter. After the arrival of the recharge water, the nitrate concentrations from the lysimeters and well at YVUZ-1 decreased to less than 1 milligram per liter, with the exception of samples collected at 205.5 feet, which did not exceed 4.12 milligrams per liter. The decrease in nitrate concentrations after artificial recharge indicated that the rising water table did not result in an increase of nitrates below YVUZ-1. At YVUZ-2, nitrate concentrations ranged between 12 to 479 milligrams per liter. The highest nitrate concentrations were at 92 feet below land surface and were almost seven times that of samples collected from a nearby septic tank. Nitrate concentrations from the lysimeter at 273 feet below land surface increased from 6 to almost 58 milligrams per liter after it was saturated by the rising water table in December 2007. These increases could be the result of the mobilization of high-nitrate water from regional sources of septic effluent after saturation, or the result of high-nitrate water present at the top of the water table that may be diluted deeper in the aquifer. Nitrate concentrations in groundwater from five nearby production wells and from both monitoring wells were less than 5 milligrams per liter before artificial recharge started. Nitrate concentrations decreased to less than 3 milligrams per liter in three of the production wells and the monitoring well below the recharge ponds after artificial recharge. Dissolved organic carbon concentrations were measured in the recharge water and groundwater because of the potential for dissolved organic carbon to react with chlorine to form trihalomethanes during the water-treatment process. The dissolved organic carbon concentrations of the recharge water were 3.1 milligrams per liter or less, and dissolved organic carbon concentrations of the groundwater were less than 1 milligram per liter. Even though recharge water was present in some of the wells by September 2008, the concentrations of both dissolved organic carbon and trihalomethane formation potential in the groundwater did not increase. Interpretation of these data suggests that the dissolved organic carbon from the recharge water is altered or metabolized in the unsaturated zone, either by absorption to the grain particles in the soil or by microbiological processes.

California

Geochemical control of microbial Fe(III) reduction potential in wetlands: Comparison of the rhizosphere to non-rhizosphere soil

We compared the reactivity and microbial reduction potential of Fe(III) minerals in the rhizosphere and non-rhizosphere soil to test the hypothesis that rapid Fe(III) reduction rates in wetland soils are explained by rhizosphere processes. The rhizosphere was defined as the area immediately adjacent to a root encrusted with Fe(III)-oxides or Fe plaque, and non-rhizosphere soil was 0.5 cm from the root surface. The rhizosphere had a significantly higher percentage of poorly crystalline Fe (66??7%) than non-rhizosphere soil (23??7%); conversely, non-rhizosphere soil had a significantly higher proportion of crystalline Fe (50??7%) than the rhizosphere (18??7%, P<0.05 in all cases). The percentage of poorly crystalline Fe(III) was significantly correlated with the percentage of FeRB (r=0.76), reflecting the fact that poorly crystalline Fe(III) minerals are labile with respect to microbial reduction. Abiotic reductive dissolution consumed about 75% of the rhizosphere Fe(III)-oxide pool in 4 h compared to 23% of the soil Fe(III)-oxide pool. Similarly, microbial reduction consumed 75-80% of the rhizosphere pool in 10 days compared to 30-40% of the non-rhizosphere soil pool. Differences between the two pools persisted when samples were amended with an electron-shuttling compound (AQDS), an Fe(III)-reducing bacterium (Geobacter metallireducens), and organic carbon. Thus, Fe(III)-oxide mineralogy contributed strongly to differences in the Fe(III) reduction potential of the two pools. Higher amounts of poorly crystalline Fe(III) and possibly humic substances, and a higher Fe(III) reduction potential in the rhizosphere compared to the non-rhizosphere soil, suggested the rhizosphere is a site of unusually active microbial Fe cycling. The results were consistent with previous speculation that rapid Fe cycling in wetlands is due to the activity of wetland plant roots. ?? 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

FEMS Microbiology Ecology

Optimization of a sample processing protocol for recovery of Bacillus anthracis spores from soil

Following a release of Bacillus anthracis spores into the environment, there is a potential for lasting environmental contamination in soils. There is a need for detection protocols for B. anthracis in environmental matrices. However, identification of B. anthracis within a soil is a difficult task. Processing soil samples helps to remove debris, chemical components, and biological impurities that can interfere with microbiological detection. This study aimed to optimize a previously used indirect processing protocol, which included a series of washing and centrifugation steps. Optimization of the protocol included: identifying an ideal extraction diluent, variation in the number of wash steps, variation in the initial centrifugation speed, sonication and shaking mechanisms. The optimized protocol was demonstrated at two laboratories in order to evaluate the recovery of spores from loamy and sandy soils. The new protocol demonstrated an improved limit of detection for loamy and sandy soils over the non-optimized protocol with an approximate matrix limit of detection at 14 spores/g of soil. There were no significant differences overall between the two laboratories for either soil type, suggesting that the processing protocol will be robust enough to use at multiple laboratories while achieving comparable recoveries.

Journal of Microbiological Methods

Arsenic, microbes and contaminated aquifers

The health of tens of millions of people world-wide is at risk from drinking arsenic-contaminated well water. In most cases this arsenic occurs naturally within the sub-surface aquifers, rather than being derived from identifiable point sources of pollution. The mobilization of arsenic into the aqueous phase is the first crucial step in a process that eventually leads to human arsenicosis. Increasing evidence suggests that this is a microbiological phenomenon.

Trends in Microbiology

Bacterial Cycling of Methyl Halides

This chapter focuses on the monohalogenated methanes methyl chloride (MeCl) and methyl bromide (MeBr), their natural and anthropogenic sources, and their degradation by microorganisms, specifically by aerobic bacteria that can use MeBr and MeCl as sole source of carbon and energy. The biogeochemical cycle of methyl halides and the microbiology, biochemistry, genetics, and biotechnological potential of methyl halide-degrading microorganisms are discussed in the chapter. Methyl halides are the dominant halocarbons in the atmosphere. They play an important role in regulating stratospheric ozone concentrations and global warming as well, two factors governing planetary habitability. The monohalomethanes—methyl chloride (MeCl), methyl bromide (MeBr), and methyl iodide (MeI)—are trace gases in the atmosphere with average tropospheric-mixing ratios of 600, 10, and 2 parts per trillion (ppt), respectively. However, methyl halides are radiatively active and hence contribute to global warming by absorbing radiation in the infrared region. This is evident in their elevated global warming potential (GWP), a value calculated to quantify each compound's warming effect (on a mass basis) relative to the same mass of CO 2 . Compounds with high GWP including those like methyl halides with low concentrations may have considerable impact on atmospheric warming when compared with other “greenhouse” gases with low GWP.

Advances in Applied Microbiology

Microbial community composition of a hydrocarbon reservoir 40 years after a CO2 enhanced oil recovery flood

Injecting CO 2 into depleted oil reservoirs to extract additional crude oil is a common enhanced oil recovery (CO 2 -EOR) technique. However, little is known about how in situ microbial communities may be impacted by CO 2 flooding, or if any permanent microbiological changes occur after flooding has ceased. Formation water was collected from an oil field that was flooded for CO 2 -EOR in the 1980s, including samples from areas affected by or outside of the flood region, to determine the impacts of CO 2 -EOR on reservoir microbial communities. Archaea, specifically methanogens, were more abundant than bacteria in all samples, while identified bacteria exhibited much greater diversity than the archaea. Microbial communities in CO 2 -impacted and non-impacted samples did not significantly differ (ANOSIM: Statistic R = -0.2597, significance = 0.769). However, several low abundance bacteria were found to be significantly associated with the CO 2 -affected group; very few of these species are known to metabolize CO 2 or are associated with CO 2 -rich habitats. Although this study had limitations, on a broad scale, either the CO 2 flood did not impact the microbial community composition of the target formation, or microbial communities in affected wells may have reverted back to pre-injection conditions over the ca. 40 years since the CO 2 -EOR.

Louisiana

Pilot- and bench-scale testing of faecal indicator bacteria survival in marine beach sand near point sources

Aim: Factors affecting faecal indicator bacteria (FIB) and pathogen survival/persistence in sand remain largely unstudied. This work elucidates how biological and physical factors affect die-off in beach sand following sewage spills. Methods and Results: Solar disinfection with mechanical mixing was pilot-tested as a disinfection procedure after a large sewage spill in Los Angeles. Effects of solar exposure, mechanical mixing, predation and/or competition, season, and moisture were tested at bench scale. First-order decay constants for Escherichia coli ranged between -0??23 and -1??02 per day, and for enterococci between -0??5 and -1??0 per day. Desiccation was a dominant factor for E. coli but not enterococci inactivation. Effects of season were investigated through a comparison of experimental results from winter, spring, and fall. Conclusions: Moisture was the dominant factor controlling E. coli inactivation kinetics. Initial microbial community and sand temperature were also important factors. Mechanical mixing, common in beach grooming, did not consistently reduce bacterial levels. Significance and Impact of the Study: Inactivation rates are mainly dependent on moisture and high sand temperature. Chlorination was an effective disinfection treatment in sand microcosms inoculated with raw influent. ?? 2009 The Society for Applied Microbiology.

Journal of Applied Microbiology

Comparison of traditional and molecular analytical methods for detecting biological agents in raw and drinking water following ultrafiltration

Aims: To compare the performance of traditional methods to quantitative polymerase chain reaction (qPCR) for detecting five biological agents in large-volume drinking-water samples concentrated by ultrafiltration (UF). Methods and Results: Drinking-water samples (100 l) were seeded with Bacillus anthracis, Cryptospordium parvum, Francisella tularensis, Salmonella Typhi, and Vibrio cholerae and concentrated by UF. Recoveries by traditional methods were variable between samples and between some replicates; recoveries were not determined by qPCR. Francisella tularensis and V. cholerae were detected in all 14 samples after UF, B. anthracis was detected in 13, and C. parvum was detected in 9 out of 14 samples. Numbers found by qPCR after UF were significantly or nearly related to those found by traditional methods for all organisms except for C. parvum. A qPCR assay for S. Typhi was not available. Conclusions: qPCR can be used to rapidly detect biological agents after UF as well as traditional methods, but additional work is needed to improve qPCR assays for several biological agents, determine recoveries by qPCR, and expand the study to other areas. Significance and Impact of the Study: To our knowledge, this is the first study to compare the use of traditional and qPCR methods to detect biological agents in large-volume drinking-water samples. ?? 2009 The Society for Applied Microbiology.

Journal of Applied Microbiology

Faecal indicator bacteria enumeration in beach sand: A comparison study of extraction methods in medium to coarse sands

Aims: The absence of standardized methods for quantifying faecal indicator bacteria (FIB) in sand hinders comparison of results across studies. The purpose of the study was to compare methods for extraction of faecal bacteria from sands and recommend a standardized extraction technique. Methods and Results: Twenty-two methods of extracting enterococci and Escherichia coli from sand were evaluated, including multiple permutations of hand shaking, mechanical shaking, blending, sonication, number of rinses, settling time, eluant-to-sand ratio, eluant composition, prefiltration and type of decantation. Tests were performed on sands from California, Florida and Lake Michigan. Most extraction parameters did not significantly affect bacterial enumeration. anova revealed significant effects of eluant composition and blending; with both sodium metaphosphate buffer and blending producing reduced counts. Conclusions: The simplest extraction method that produced the highest FIB recoveries consisted of 2 min of hand shaking in phosphate-buffered saline or deionized water, a 30-s settling time, one-rinse step and a 10 : 1 eluant volume to sand weight ratio. This result was consistent across the sand compositions tested in this study but could vary for other sand types. Significance and Impact of the Study: Method standardization will improve the understanding of how sands affect surface water quality. ?? 2009 The Society for Applied Microbiology.

Journal of Applied Microbiology

Population structure, persistence, and seasonality of autochthonous Escherichia coli in temperate, coastal forest soil from a Great Lakes watershed

The common occurrence of Escherichia coli in temperate soils has previously been reported, however, there are few studies to date to characterize its source, distribution, persistent capability and genetic diversity. In this study, undisturbed, forest soils within six randomly selected 0.5 m2 exclosure plots (covered by netting of 2.3 mm2 mesh size) were monitored from March to October 2003 for E. coli in order to describe its numerical and population characteristics. Culturable E. coli occurred in 88% of the samples collected, with overall mean counts of 16 MPN g-1, ranging from <1 to 1657 (n = 66). Escherichia coli counts did not correlate with substrate moisture content, air, or soil temperatures, suggesting that seasonality were not a strong factor in population density control. Mean E. coli counts in soil samples (n = 60) were significantly higher inside than immediately outside the exclosures; E. coli distribution within the exclosures was patchy. Repetitive extragenic palindromic polymerase chain reaction (Rep-PCR) demonstrated genetic heterogeneity of E. coli within and among exclosure sites, and the soil strains were genetically distinct from animal (E. coli) strains tested (i.e. gulls, terns, deer and most geese). These results suggest that E. coli can occur and persist for extended periods in undisturbed temperate forest soils independent of recent allochthonous input and season, and that the soil E. coli populations formed a cohesive phylogenetic group in comparison to the set of fecal strains with which they were compared. Thus, in assessing E. coli sources within a stream, it is important to differentiate background soil loadings from inputs derived from animal and human fecal contamination. ?? 2005 Society for Applied Microbiology and Blackwell Publishing Ltd.

Environmental Microbiology

Assessment of fecal pollution sources in a small northern-plains watershed using PCR and phylogenetic analyses of Bacteroidetes 16S rRNA gene

We evaluated the efficacy, sensitivity, host-specificity, and spatial/temporal dynamics of human- and ruminant-specific 16S rRNA gene Bacteroidetes markers used to assess the sources of fecal pollution in a fecally impacted watershed. Phylogenetic analyses of 1271 fecal and environmental 16S rRNA gene clones were also performed to study the diversity of Bacteroidetes in this watershed. The host-specific assays indicated that ruminant feces were present in 28-54% of the water samples and in all sampling seasons, with increasing frequency in downstream sites. The human-targeted assays indicated that only 3-5% of the water samples were positive for human fecal signals, although a higher percentage of human-associated signals (19-24%) were detected in sediment samples. Phylogenetic analysis indicated that 57% of all water clones clustered with yet-to-be-cultured Bacteroidetes species associated with sequences obtained from ruminant feces, further supporting the prevalence of ruminant contamination in this watershed. However, since several clusters contained sequences from multiple sources, future studies need to consider the potential cosmopolitan nature of these bacterial populations when assessing fecal pollution sources using Bacteroidetes markers. Moreover, additional data is needed in order to understand the distribution of Bacteroidetes host-specific markers and their relationship to water quality regulatory standards. ?? 2006 Federation of European Microbiological Societies.

FEMS Microbiology Ecology

Three distinct clades of cultured heterocystous cyanobacteria constitute the dominant N2-fixing members of biological soil crusts of the Colorado Plateau, USA

The identity of the numerically dominant N2-fixing bacteria in biological soil crusts of the Colorado Plateau region and two outlying areas was determined using multiple approaches, to link the environmental diversity of nifH gene sequences to cultured bacterial isolates from the regions. Of the nifH sequence-types detected in soil crusts of the Colorado Plateau, 89% (421/473) were most closely related to nifH signature sequences from cyanobacteria of the order Nostocales. N2-fixing cyanobacterial strains were cultured from crusts and their morphotypes, 16S rRNA gene and nifH gene sequences were characterized. The numerically dominant diazotrophs in the Colorado Plateau crusts fell within three clades of heterocystous cyanobacteria. Two clades are well-represented by phylogenetically and morphologically coherent strains, corresponding to the descriptions of Nostoc commune and Scytonema hyalinum, which are widely recognized as important N2-fixing components of soil crusts. A third, previously-overlooked clade was represented by a phylogenetically coherent but morphologically diverse group of strains that encompass the morphogenera Tolypothrix and Spirirestis. Many of the strains in each of these groups contained at least two nifH copies that represent different clusters in the nifH environmental survey. ?? 2007 Federation of European Microbiological Societies.

FEMS Microbiology Ecology

Acquisition of nonspecific Bartonella strains by the northern grasshopper mouse (Onychomys leucogaster)

Rodent-associated Bartonella species are generally host-specific parasites in North America. Here evidence that Bartonella species can 'jump' between host species is presented. Northern grasshopper mice and other rodents were trapped in the western USA. A study of Bartonella infection in grasshopper mice demonstrated a high prevalence that varied from 25% to 90% by location. Bartonella infection was detected in other rodent species with a high prevalence as well. Sequence analyses of gltA identified 29 Bartonella variants in rodents, 10 of which were obtained from grasshopper mice. Among these 10, only six variants were specific to grasshopper mice, whereas four were identical to variants specific to deer mice or 13-lined ground squirrels. Fourteen of 90 sequenced isolates obtained from grasshopper mice were strains found more commonly in other rodent species and were apparently acquired from these animals. The ecological behavior of grasshopper mice may explain the occurrence of Bartonella strains in occasional hosts. The observed rate at which Bartonella jumps from a donor host species to the grasshopper mouse was directly proportional to a metric of donor host density and to the prevalence of Bartonella in the donor host, and inversely proportional to the same parameters for the grasshopper mouse. ?? 2007 Federation of European Microbiological Societies.

FEMS Microbiology Ecology

Microbial consortia of gorgonian corals from the Aleutian islands

Gorgonians make up the majority of corals in the Aleutian archipelago and provide critical fish habitat in areas of economically important fisheries. The microbial ecology of the deep-sea gorgonian corals Paragorgea arborea, Plumarella superba , and Cryogorgia koolsae was examined with culture-based and 16S rRNA gene-based techniques. Six coral colonies (two per species) were collected. Samples from all corals were cultured, and clone libraries were constructed from P. superba and C. koolsae . Cultured bacteria were dominated by the Gammaproteobacteria , especially Vibrionaceae , with other phyla comprising <6% of the isolates. The clone libraries showed dramatically different bacterial communities between corals of the same species collected at different sites, with no clear pattern of conserved bacterial consortia. Two of the clone libraries (one from each coral species) were dominated by Tenericutes , with Alphaproteobacteria dominating the remaining sequences. The other libraries were more diverse and had a more even distribution of bacterial phyla, showing more similarity between genera than within coral species. Here we report the first microbiological characterization of P. arborea, P. superba , and C. koolsae .

Alaska

Bacterial and archaeal phylogenetic diversity of a cold sulfur-rich spring on the shoreline of Lake Erie, Michigan

Studies of sulfidic springs have provided new insights into microbial metabolism, groundwater biogeochemistry, and geologic processes. We investigated Great Sulphur Spring on the western shore of Lake Erie and evaluated the phylogenetic affiliations of 189 bacterial and 77 archaeal 16S rRNA gene sequences from three habitats: the spring origin (11-m depth), bacterial-algal mats on the spring pond surface, and whitish filamentous materials from the spring drain. Water from the spring origin water was cold, pH 6.3, and anoxic (H 2 , 5.4 nM; CH 4 , 2.70 ??M) with concentrations of S 2- (0.03 mM), SO 4 2- (14.8 mM), Ca 2+ (15.7 mM), and HCO 3 - (4.1 mM) similar to those in groundwater from the local aquifer. No archaeal and few bacterial sequences were >95% similar to sequences of cultivated organisms. Bacterial sequences were largely affiliated with sulfur-metabolizing or chemolithotrophic taxa in Beta-, Gamma-, Delta-, and Epsilonproteobacteria. Epsilonproteobacteria sequences similar to those obtained from other sulfidic environments and a new clade of Cyanobacteria sequences were particularly abundant (16% and 40%, respectively) in the spring origin clone library. Crenarchaeota sequences associated with archaeal-bacterial consortia in whitish filaments at a German sulfidic spring were detected only in a similar habitat at Great Sulphur Spring. This study expands the geographic distribution of many uncultured Archaea and Bacteria sequences to the Laurentian Great Lakes, indicates possible roles for epsilonproteobacteria in local aquifer chemistry and karst formation, documents new oscillatorioid Cyanobacteria lineages, and shows that uncultured, cold-adapted Crenarchaeota sequences may comprise a significant part of the microbial community of some sulfidic environments. Copyright ?? 2009, American Society for Microbiology. All Rights Reserved.

Applied and Environmental Microbiology