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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

Effects of aquifer storage and recovery activities on water quality in the Little Arkansas River and Equus Beds Aquifer, south-central Kansas, 2011–14

The Equus Beds aquifer in south-central Kansas is aprimary water source for the city of Wichita. The Equus Beds aquifer storage and recovery (ASR) project was developed to help the city of Wichita meet increasing current (2016) and future water demands. The Equus Beds ASR project pumps water out of the Little Arkansas River during above-base flow conditions, treats it using drinking-water quality standards as a guideline, and recharges it into the Equus Beds aquifer for later use. Phase II of the Equus Beds ASR project currently (2016) includes a river intake facility and a surface-water treatment facility with a 30 million gallon per day capacity. Water diverted from the Little Arkansas River is delivered to an adjacent presedimentation basin for solids removal. Subsequently, waste from the surface-water treatment facility and the presedimentation basin is returned to the Little Arkansas River through a residuals return line. The U.S. Geological Survey, in cooperation with the city of Wichita, developed and implemented a hydrobiological monitoring program as part of the ASR project to characterize and quantify the effects of aquifer storage and recovery activities on the Little Arkansas River and Equus Beds aquifer water quality. Data were collected from 2 surface-water sites (one upstream and one downstream from the residuals return line), 1 residuals return line site, and 2 groundwater well sites (each having a shallow and deep part): the Little Arkansas River upstream from the ASR facility near Sedgwick, Kansas (upstream surface-water site 375350097262800), about 0.03 mile (mi) upstream from the residuals return line site; the Little Arkansas River near Sedgwick, Kans. (downstream surface-water site 07144100), about 1.68 mi downstream from the residuals return line site; discharge from the Little Arkansas River ASR facility near Sedgwick, Kansas (residuals return line site 375348097262800); 25S 01 W 07BCCC01 SMW–S11 near CW36 (MW–7 shallow groundwater well site 375327097285401); 25S01 W 07BCCC02 DMW–S10 near CW36 (MW–7 deep groundwater well site 375327097285402); 25S 01W 07BCCA01 SMW–S13 near CW36 (MW–8 shallow groundwater well site 375332097284801); and 25S 01W 07BCCA02 DMW–S14 near CW36 (MW–8 deep groundwater well site 375332097284802). The U.S. Geological Survey, in cooperation with the city of Wichita, assessed the effects of the ASR Phase II facility residuals return line discharges on stream quality of the Little Arkansas River by measuring continuous physicochemical properties and collecting discrete water-quality and sediment samples for about 2 years pre- (January 2011 through April 2013) and post-ASR (May 2013 through December 2014) Phase II facility operation upstream and downstream from the ASR Phase II facility. Additionally, habitat variables were quantified and macroinvertebrate and fish communities were sampled upstream and downstream from the ASR Phase II facility during the study period. To assess the effects of aquifer recharge on Equus Beds groundwater quality, continuous physicochemical properties were measured and discrete water-quality samples were collected before and during the onset of Phase II aquifer recharge in two (shallow and deep) groundwater wells. Little Arkansas River streamflow was about 10 times larger after the facility began operating because of greater rainfall. Residuals return line release volumes were a very minimal proportion (0.06 percent) of downstream streamflow volume during the months the ASR facility was operating. Upstream and downstream continuously measured water temperature and dissolved oxygen median differences were smaller post-ASR than pre-ASR. Turbidity generally was smaller at the downstream site throughout the study period and decreased at both sites after the ASR Phase II facility began discharging despite a median residuals return line turbidity that was about an order of magnitude larger than the median turbidity at the downstream site. Upstream and downstream continuously measured turbidity median differences were larger post-ASR than pre-ASR. Median post-ASR continuously measured nitrite plus nitrate and continuously computed total suspended solids and suspended-sediment concentrations were smaller than pre-ASR likely because of higher streamflows and dilution; whereas, median continuously computed dissolved and total organic carbon concentrations were larger likely because of higher streamflows and runoff conditions. None of the discretely measured water-quality constituents (dissolved and suspended solids, primary ions, suspended sediment, nutrients, carbon, trace elements, viral and bacterial indicators, and pesticides) in surface water were significantly different between the upstream and downstream sites after the ASR Phase II facility began discharging; however, pre-ASR calcium, sodium, hardness, manganese, and arsenate concentrations were significantly larger at the upstream site, which indicates that some water-quality conditions at the upstream and downstream sites were more similar post-ASR. Most of the primary constituents that make up dissolved solids decreased at both sites after the ASR Phase II facility began operation. Discretely collected total suspended solids concentrations were similar between the upstream and downstream sites before the facility began operating but were about 27 percent smaller at the downstream site after the facility began operating, despite the total suspended solids concentrations in the residuals return line being 15 times larger than the downstream site. Overall habitat scores were indicative of suboptimal conditions upstream and downstream from the ASR Phase II facility throughout the study period. Substrate fouling and sediment deposition mean scores indicated marginal conditions at the upstream and downstream sites during the study period, demonstrating that sediment deposition was evident pre- and post-ASR and no substantial changes in these habitat characteristics were noted after the ASR Phase II facility began discharging. Macroinvertebrate community composition (evaluated using functional feeding, behavioral, and tolerance metrics) generally was similar between sites during the study period. Fewer macroinvertebrate metrics were significant between the upstream and downstream sites post-ASR (6) than pre-ASR (14), which suggests that macroinvertebate communities were more similar after the ASR facility began discharging. Upstream-downstream comparisons in macroinvertebrate aquatic-life-support metrics had no significant differences for the post-ASR time period and neither site was fully supporting for any of the Kansas Department of Health and Environment aquatic-life-support metrics (Macroinvertebrate Biotic Index; Kansas Biotic Index with tolerances for nutrients and oxygen-demanding substances; Ephemeroptera, Plecoptera, and Trichoptera [EPT] richness; and percentage of EPT species). Overall, using macroinvertebrate aquatic life-support criteria from the Kansas Department of Health and Environment, upstream and downstream sites were classified as partially supporting before and after the onset of ASR facility operations. Fish community trophic status and tolerance groups generally were similar among sites during the study period. Fish community Little Arkansas River Basin Index of Biotic Integrity scores at the upstream and downstream sites were indicative of fair-to-good conditions before the facility began operating and decreased to fair conditions after the facility began operating. Groundwater physicochemical changes concurrent with the beginning of recharge operations at the Sedgwick basin were more pronounced in shallow groundwater. No constituent concentrations in the pre-recharge period in comparison to the post-recharge period increased to concentrations exceeding drinking water regulations; however, nitrate decreased significantly from a pre-recharge exceedance of the U.S. Environmental Protection Agency maximum contaminant level to a post recharge nonexceedance. Shallow groundwater chemical concentrations or rates of detection increased after artificial recharge began for the ions potassium, chloride, and fluoride; phosphorus and organic carbon species; trace elements barium, manganese, nickel, arsenate, arsenic, and boron; agricultural pesticides atrazine, metolachlor, metribuzin, and simazine; organic disinfection byproducts bromodichloromethane and trichloromethane; and gross beta levels. Additionally, water temperature, and pH were larger after recharge began; and total solids and slime-forming bacteria concentrations and densities were smaller. Total solids, nitrate, and selenium significantly decreased; and potassium, chloride, nickel, arsenic, fluoride, phosphorus and carbon species, and gross beta levels significantly increased in shallow groundwater after artificial recharge. Results of biological activity reaction tests indicated that water quality microbiology was different before and after artificial recharge began; at times, these differences may lead to changes in dominant bacterial populations that, in turn, may lead to formation and expansion in populations that may cause bioplugging and other unwanted effects. Calcite, iron (II) hydroxide, hydroxyapatite, and similar minerals, had shifts in saturation indices that generally were from undersaturation toward equilibrium and, in some cases, toward oversaturation. These shifts toward neutral saturation indices might suggest reduced weathering of the minerals present in the Equus Beds aquifer. Chemical weathering in the shallow parts of the aquifer may be accelerated because of the increased water temperatures and the system is more vulnerable to clogged pores and mineral dissolution as the equilibrium state is affected by recharge and withdrawal. When oversaturation is indicated for iron minerals, plugging of aquifer materials may happen.

Kansas

Exchange of carbapenem-resistant Escherichia coli Sequence Type 38 intercontinentally and among wild bird, human, and environmental niches

Carbapenem-resistant Enterobacteriaceae (CRE) are a global threat to human health and are increasingly being isolated from nonclinical settings. OXA-48-producing Escherichia coli sequence type 38 (ST38) is the most frequently reported CRE type in wild birds and has been detected in gulls or storks in North America, Europe, Asia, and Africa. The epidemiology and evolution of CRE in wildlife and human niches, however, remains unclear. We compared wild bird origin E. coli ST38 genome sequences generated by our research group and publicly available genomic data derived from other hosts and environments to (i) understand the frequency of intercontinental dispersal of E. coli ST38 clones isolated from wild birds, (ii) more thoroughly measure the genomic relatedness of carbapenem-resistant isolates from gulls sampled in Turkey and Alaska, USA, using long-read whole-genome sequencing and assess the spatial dissemination of this clone among different hosts, and (iii) determine whether ST38 isolates from humans, environmental water, and wild birds have different core or accessory genomes (e.g., antimicrobial resistance genes, virulence genes, plasmids) which might elucidate bacterial or gene exchange among niches. Our results suggest that E. coli ST38 strains, including those resistant to carbapenems, are exchanged between humans and wild birds, rather than separately maintained populations within each niche. Furthermore, despite close genetic similarity among OXA-48-producing E. coli ST38 clones from gulls in Alaska and Turkey, intercontinental dispersal of ST38 clones among wild birds is uncommon. Interventions to mitigate the dissemination of antimicrobial resistance throughout the environment (e.g., as exemplified by the acquisition of carbapenem resistance by birds) may be warranted.

Applied and Environmental Microbiology

Effect of growth conditions and staining procedure upon the subsurface transport and attachment behaviors of a groundwater protist

The transport and attachment behaviors of Spumella guttula (Kent), a nanoflagellate (protist) found in contaminated and uncontaminated aquifer sediments in Cape Cod, Mass., were assessed in flowthrough and static columns and in a field injection-and-recovery transport experiment involving an array of multilevel samplers. Transport of S. guttula harvested from low-nutrient (10 mg of dissolved organic carbon per liter), slightly acidic, granular (porous) growth media was compared to earlier observations involving nanoflagellates grown in a traditional high-nutrient liquid broth. In contrast to the highly retarded (retardation factor of ∼3) subsurface transport previously reported for S. guttula , the peak concentration of porous-medium-grown S. guttula traveled concomitantly with that of a conservative (bromide) tracer. About one-third of the porous-medium-grown nanoflagellates added to the aquifer were transported at least 2.8 m downgradient, compared to only ∼2% of the broth-grown nanoflagellates. Flowthrough column studies revealed that a vital (hydroethidine [HE]) staining procedure resulted in considerably less attachment (more transport) of S. guttula in aquifer sediments than did a staining-and-fixation procedure involving 4′,6′-diamidino-2-phenylindole (DAPI) and glutaraldehyde. The calculated collision efficiency (∼10 −2 for porous-medium-grown, DAPI-stained nanoflagellates) was comparable to that observed earlier for the indigenous community of unattached groundwater bacteria that serve as prey. The attachment of HE-labeled S. guttula onto aquifer sediment grains was independent of pH (over the range from pH 3 to 9) suggesting a primary attachment mechanism that may be fundamentally different from that of their prey bacteria, which exhibit sharp decreases in fractional attachment with increasing pH. The high degree of mobility of S. guttula in the aquifer sediments has important ecological implications for the protistan community within the temporally changing plume of organic contaminants in the Cape Cod aquifer.

Applied and Environmental Microbiology

Characteristics of water quality and streamflow, Passaic River basin above Little Falls, New Jersey

The findings of a problem-oriented river-system investigation of the water-quality and streamflow characteristics of the Passaic River above Little Falls, N.J. (drainage area 762 sq mi) are described. Information on streamflow duration, time-of-travel measurements, and analyses of chemical, biochemical, and physical water quality are summarized. This information is used to define relations between water quality, streamflow, geology, and environmental development in the basin's hydrologic system. The existence, nature, and magnitude of long-term trends in stream quality--as measured by dissolved solids, chloride, dissolved oxygen, biochemical oxygen demand, ammonia, nitrate, and turbidity--and in streamflow toward either improvement or deterioration are appraised at selected sites within the river system. The quality of streams in the upper Passaic River basin in northeastern New Jersey is shown to be deteriorating with time. For example, biochemical oxygen demand, an indirect measure of organic matter in a stream, is increasing at most stream-quality sampling sites. Similarly, the dissolved-solids content, a measure of inorganic matter, also is increasing. These observations suggest that the Passaic River system is being used more and more as a medium for the disposal of industrial and municipal waste waters. Dissolved oxygen, an essential ingredient for the natural purification of streams receiving waste discharges, is undersaturated (that is, below theoretical solubility levels) at all sampling sites and is decreasing with time at most sites. This is another indication of the general deterioration of stream quality in the upper basin. It also indicates that the ability of the river system to receive, transport, and assimilate wastes, although exceeded now only for short periods during the summer months, may be exceeded more continually in the future if present trends hold. Decreasing ratios of ammonia to nitrate in a downstream direction on the main stem Passaic River suggests that nitrification (the biochemical conversion of ammonia to nitrate) as well as microbiological decomposition of organic matter (waste waters) is contributing to the continued and increasing undersaturation of dissolved oxygen in the river system. Passaic River streams are grouped into five general regions of isochemical quality on the basis of predominant constituents and dissolved-solids content during low flows. The predominant cations in all but one region are calcium and magnesium (exceeding 50 percent of total cations) ; in that region, where man's activities probably have altered the natural stream waters, the percentage of sodium and potassium equals that of calcium and magnesium. In two of the five regions, the predominant anion is bicarbonate; a combination of sulfate, chloride, and nitrate is predominant in the other three regions. Dissolved-solids content during low flows generally ranges from 100 to 600 milligrams per liter. Several time-of-travel measurements within the basin are reported. These data provide reasonable estimates of the time required for soluble contaminants to pass through particular parts of the river system. For example, the peak concentration of a contaminant injected into the river system at Chatham during extreme low flow would be expected to travel to Little Falls, about 31 miles, in about 13 days; but at medium flow, in about 5 days.

Water Supply Paper

Bacterial genomics reveal the complex epidemiology of an emerging pathogen in arctic and boreal ungulates

Northern ecosystems are currently experiencing unprecedented ecological change, largely driven by a rapidly changing climate. Pathogen range expansion, and emergence and altered patterns of infectious disease, are increasingly reported in wildlife at high latitudes. Understanding the causes and consequences of shifting pathogen diversity and host-pathogen interactions in these ecosystems is important for wildlife conservation, and for indigenous populations that depend on wildlife. Among the key questions are whether disease events are associated with endemic or recently introduced pathogens, and whether emerging strains are spreading throughout the region. In this study, we used a phylogenomic approach to address these questions of pathogen endemicity and spread for Erysipelothrix rhusiopathiae , an opportunistic multi-host bacterial pathogen associated with recent mortalities in arctic and boreal ungulate populations in North America. We isolated E. rhusiopathiae from carcasses associated with large-scale die-offs of muskoxen in the Canadian Arctic Archipelago, and from contemporaneous mortality events and/or population declines among muskoxen in northwestern Alaska and caribou and moose in western Canada. Bacterial genomic diversity differed markedly among these locations; minimal divergence was present among isolates from muskoxen in the Canadian Arctic, while in caribou and moose populations, strains from highly divergent clades were isolated from the same location, or even from within a single carcass. These results indicate that mortalities among northern ungulates are not associated with a single emerging strain of E. rhusiopathiae , and that alternate hypotheses need to be explored. Our study illustrates the value and limitations of bacterial genomic data for discriminating between ecological hypotheses of disease emergence, and highlights the importance of studying emerging pathogens within the broader context of environmental and host factors.

Frontiers in Microbiology

Occurrence and distribution of microbiological contamination and enteric viruses in shallow ground water in Baltimore and Harford counties, Maryland

The U.S. Geological Survey, in cooperation with the Maryland Department of the Environment and the Wisconsin State Laboratory of Hygiene, conducted a study to characterize the occurrence and distribution of viral contamination in small (withdrawing less than 10,000 gallons per day) public water-supply wells screened in the shallow aquifer in the Piedmont Physiographic Province in Baltimore and Harford Counties, Maryland. Two hundred sixty-three small public water-supply wells were in operation in these counties during the spring of 2000. Ninety-one of these sites were selected for sampling using a methodology that distributed the samples evenly over the population and the spatial extent of the study area. Each site, and its potential susceptibility to microbiological contamination, was evaluated with regard to hole depth, casing interval, and open interval. Each site was evaluated using characteristics such as on-site geology and on-site land use.Samples were collected by pumping between 200 and 400 gallons of untreated well water through an electropositive cartridge filter. Water concentrates were subjected to cell-culture assay for the detection of culturable viruses and reverse-transcription polymerase chain reaction/gene probe assays to detect viral ribonucleic acid; grab samples were analyzed for somatic and male-specific coliphages, Bacteroides fragilis, Clostridium perfringens, enterococci, Escherichia coli, total coliforms, total oxidized nitrogen, nitrite, organic nitrogen, total phosphate, ortho-phosphate, calcium, magnesium, sodium, potas-sium, chloride, sulfate, iron, acid-neutralizing capacity, pH, specific conductance, temperature, and dissolved oxygen.One sample tested positive for the presence of the ribonucleic acid of rotavirus through poly-merase chain-reaction analysis. Twenty-nine per-cent of the samples (26 of 90) had bacterial con-tamination. About 7 percent of the samples (6 of 90) were contaminated with either male-specific coliphage, somatic coliphage, or bacteriophages of Bacteroides fragilis. About 3 percent of the sam-ples (3 of 87) had oxidized nitrogen concentra-tions that exceeded the U.S. Environmental Protection Agency?s Maximum Contaminant Level of 10.0 milligrams per liter. A statistical analysis showed that no significant relation exists between the presence of bacteria or coliphage and all variables, except the mean temperature of the water sample as measured in the field. Additionally, the concentration of total coliform bacteria had a statistically significant, moderately strong cor-relation with the concentration of sulfate and sample pH as measured at the U.S. Geological Survey National Water-Quality Laboratory in Denver, Colorado.

Water-Resources Investigations Report

Ground-water quality and discharge to Chincoteague and Sinepuxent Bays adjacent to Assateague Island National Seashore, Maryland

The U.S. Geological Survey, in cooperation with the Maryland Department of the Environment and the Wisconsin State Laboratory of Hygiene, conducted a study to characterize the occurrence and distribution of viral contamination in small (withdrawing less than 10,000 gallons per day) public water-supply wells screened in the shallow aquifer in the Piedmont Physiographic Province in Baltimore and Harford Counties, Maryland. Two hundred sixty-three small public water-supply wells were in operation in these counties during the spring of 2000. Ninety-one of these sites were selected for sampling using a methodology that distributed the samples evenly over the population and the spatial extent of the study area. Each site, and its potential susceptibility to microbiological contamination, was evaluated with regard to hole depth, casing interval, and open interval. Each site was evaluated using characteristics such as on-site geology and on-site land use.Samples were collected by pumping between 200 and 400 gallons of untreated well water through an electropositive cartridge filter. Water concentrates were subjected to cell-culture assay for the detection of culturable viruses and reverse-transcription polymerase chain reaction/gene probe assays to detect viral ribonucleic acid; grab samples were analyzed for somatic and male-specific coliphages, Bacteroides fragilis, Clostridium perfringens, enterococci, Escherichia coli, total coliforms, total oxidized nitrogen, nitrite, organic nitrogen, total phosphate, ortho-phosphate, calcium, magnesium, sodium, potas-sium, chloride, sulfate, iron, acid-neutralizing capacity, pH, specific conductance, temperature, and dissolved oxygen.One sample tested positive for the presence of the ribonucleic acid of rotavirus through poly-merase chain-reaction analysis. Twenty-nine per-cent of the samples (26 of 90) had bacterial con-tamination. About 7 percent of the samples (6 of 90) were contaminated with either male-specific coliphage, somatic coliphage, or bacteriophages of Bacteroides fragilis. About 3 percent of the sam-ples (3 of 87) had oxidized nitrogen concentra-tions that exceeded the U.S. Environmental Protection Agency?s Maximum Contaminant Level of 10.0 milligrams per liter. A statistical analysis showed that no significant relation exists between the presence of bacteria or coliphage and all variables, except the mean temperature of the water sample as measured in the field. Additionally, the concentration of total coliform bacteria had a statistically significant, moderately strong cor-relation with the concentration of sulfate and sample pH as measured at the U.S. Geological Survey National Water-Quality Laboratory in Denver, Colorado.

Water-Resources Investigations Report

Epidemiology of a Salmonella enterica subsp. Enterica serovar Typhimurium strain associated with a songbird outbreak.

Salmonella enterica subsp. enterica serovar Typhimurium is responsible for the majority of salmonellosis cases worldwide. This Salmonella serovar is also responsible for die-offs in songbird populations. In 2009, there was an S. Typhimurium epizootic reported in pine siskins in the eastern United States. At the time, there was also a human outbreak with this serovar that was associated with contaminated peanuts. As peanuts are also used in wild-bird food, it was hypothesized that the pine siskin epizootic was related to this human outbreak. A comparison of songbird and human S. Typhimurium pulsed-field gel electrophoresis (PFGE) patterns revealed that the epizootic was attributed not to the peanut-associated strain but, rather, to a songbird strain first characterized from an American goldfinch in 1998. This same S. Typhimurium strain (PFGE type A3) was also identified in the PulseNet USA database, accounting for 137 of 77,941 total S. Typhimurium PFGE entries. A second molecular typing method, multiple-locus variable-number tandem-repeat analysis (MLVA), confirmed that the same strain was responsible for the pine siskin epizootic in the eastern United States but was distinct from a genetically related strain isolated from pine siskins in Minnesota. The pine siskin A3 strain was first encountered in May 2008 in an American goldfinch and later in a northern cardinal at the start of the pine siskin epizootic. MLVA also confirmed the clonal nature of S. Typhimurium in songbirds and established that the pine siskin epizootic strain was unique to the finch family. For 2009, the distribution of PFGE type A3 in passerines and humans mirrored the highest population density of pine siskins for the East Coast.

Applied and Environmental Microbiology

Chemical and microbiological water quality of subsurface agricultural drains during a field trial of liquid dairy manure effluent application rate and varying tillage practices, Upper Tiffin Watershed, southeastern Michigan

A field trial was done in the Upper Tiffin River Watershed, in southeastern Michigan, to determine the influence of liquid dairy manure effluent (LDME) management practices on the quality of agricultural subsurface-drain water. Samples from subsurface drains were analyzed for nutrients, fecal-coliform and Escherichia coli ( E. coli ) bacteria, antibiotics, chemicals typically detected in wastewater, and the occurrence of genes indicating the presence of shiga-toxin-producing E. coli , or of bovine-specific Bacteroidetes bacteria. Samples were collected from November 2, 2006, to March 20, 2007, from eight subsurface drains under field plots that received no LDME and no tillage (controls) or received 4,000 or 8,000 gallons per acre (gal/acre) of LDME and either no tillage or two different types of tillage. The two types of tillage tested were (1) ground-driven, rotary, subsurface cultivation and (2) rolling-tine aeration. Samples were collected before LDME application and at 4 hours, and 1, 2, 6, 7, and 14 days post-application. Nutrient concentrations were high in subsurface-drain water throughout the field-trial period and could not be attributed to the field-trial LDME application. Of the 59 drain-water samples, including those collected before LDME application and control samples for each date, 56 had concentrations greater than the U.S. Environmental Protection Agency (USEPA), Ecoregion VI recommended surface-water criterion for total phosphorus, and all samples had concentrations greater than the recommended total nitrogen criterion. Nitrate + nitrite nitrogen concentration exceeded 20 milligrams per liter for every sample and contributed most to the total nitrogen concentrations. Substantial increases in drain-water concentrations of organic and ammonia nitrogen and total phosphorus were found for all treatments, including controls, at 14 days post-application after 0.84 inch of rainfall over 2 days. E. coli concentrations exceeded the USEPA recreational-water-quality single-sample criterion of 235 colony forming units per 100 milliliters in only 3 of 56 samples. Of these three samples, two were collected within 1 day post-LDME application from the treatment receiving 8,000 gal/acre LDME with no tillage (NT8000). The third sample was from the rolling-tine aerator treatment with 4,000 gal/acre LDME application rate after the first significant rainfall. Two wastewater chemicals and two bacterial genes ( eae A and stx 1) detected in the LDME, but absent in field blank or pre-application samples, were detected in the 4-hour or 1-day postapplication NT8000 samples. No LDME-associated chemicals were detected in later samples from the NT8000 treatment, and none were detected in samples from other treatments after the first significant rainfall. Results of this field trial were somewhat equivocal with respect to the influence of LDME concentration and tillage practices on subsurface-drain water quality, both immediately after LDME application and in the longer term, after significant rainfall. Interpretation of study findings is limited by the fact that treatments were not replicated, and flow rate or discharge from the subsurface drains was not measured. Nevertheless, study results provide useful information about nutrient and bacteria concentrations in subsurface drains during the non-growing season. In addition, study results demonstrate some potential for the use of chemical and microbiological indicators of LDME transport to subsurface drains.

Michigan

Factors affecting groundwater quality in the Valley and Ridge aquifers, eastern United States, 1993-2002

Chemical and microbiological analyses of water from 230 wells and 35 springs in the Valley and Ridge Physiographic Province, sampled between 1993 and 2002, indicated that bedrock type (carbonate or siliciclastic rock) and land use were dominant factors influencing groundwater quality across a region extending from northwestern Georgia to New Jersey. The analyses included naturally occurring compounds (major mineral ions and radon) and anthropogenic contaminants [pesticides and volatile organic compounds (VOCs)], and contaminants, such as nitrate and bacteria, which commonly increase as a result of human activities. Natural factors, such as topographic position and the mineral composition of underlying geology, act to produce basic physical and geochemical conditions in groundwater that are reflected in physical properties, such as pH, temperature, specific conductance, and alkalinity, and in chemical concentrations of dissolved oxygen, radon, and major mineral ions. Anthropogenic contaminants were most commonly found in water from wells and springs in carbonate-rock aquifers. Nitrate concentrations exceeded U.S. Environmental Protection Agency maximum contaminant levels in 12 percent of samples, most of which were from carbonate-rock aquifers. Escherichia coli (E. coli), pesticide, and VOC detection frequencies were significantly higher in samples from sites in carbonate-rock aquifers. Naturally occurring elements, such as radon, iron, and manganese, were found in higher concentrations in siliciclastic-rock aquifers. Radon levels exceeded the proposed maximum contaminant level of 300 picocuries per liter in 74 percent of the samples, which were evenly distributed between carbonate- and siliciclastic-rock aquifers. The land use in areas surrounding wells and springs was another significant explanatory variable for the occurrence of anthropogenic compounds. Nitrate and pesticide concentrations were highest in samples collected from sites in agricultural areas and lowest in samples collected from sites in undeveloped areas. Volatile organic compounds were detected most frequently and in highest concentrations in samples from sites in urban areas, and least frequently in agricultural and undeveloped areas. No volatile organic compound concentrations and concentrations from only one pesticide, dieldrin, exceeded human-health benchmarks.

Alabama;Georgia;Tennessee;North Carolina;Virginia;

Possible sources of nitrate in ground water at swine licensed-managed feeding operations in Oklahoma, 2001

Samples collected and analyzed by the Oklahoma Department of Agriculture, Food, and Forestry from 1999 to 2001 determined that nitrate exceeded the U.S. Environmental Protection Agency maximum contaminant level for public drinking-water supplies of 10 milligrams per liter as nitrogen in 79 monitoring wells at 35 swine licensed-managed feeding operations (LMFO) in Oklahoma. The LMFOs are located in rural agricultural settings where long-term agriculture has potentially affected the ground-water quality in some areas. Land use prior to the construction of the LMFOs was assessed to evaluate the types of agricultural land use within a 500-meter radius of the sampled wells. Chemical and microbiological techniques were used to determine the possible sources of nitrate in water sampled from 10 wastewater lagoons and 79 wells. Samples were analyzed for dissolved major ions, dissolved trace elements, dissolved nutrients, nitrogen isotope ratios of nitrate and ammonia, wastewater organic compounds, and fecal coliform bacteria. Bacteria ribotyping analysis was done on selected samples to identify possible specific animal sources. A decision process was developed to identify the possible sources of nitrate. First, nitrogen isotope ratios were used to define sources as animal, mixed animal and fertilizer, or fertilizer. Second, wastewater organic compound detections, nitrogen-isotope ratios, fecal coliform bacteria detections, and ribotyping were used to refine the identification of possible sources as LFMO waste, fertilizer, or unidentified animal or mixtures of these sources. Additional evidence provided by ribotyping and wastewater organic compound data can, in some cases, specifically indicate the animal source. Detections of three or more wastewater organic compounds that are indicators of animal sources and detections of fecal coliform bacteria provided additional evidence of an animal source. LMFO waste was designated as a possible source of nitrate in water from 10 wells. The source of waste in water from five of those wells was determined through ribotyping, and the source of waste in water from the remaining five wells was determined by detections of three or more animal-waste compounds in the well samples. LMFO waste in the water from wells with unidentified animal source of nitrate does not indicate that LMFO waste was not the source, but indicated that multiple animal sources, including LMFO waste, may be the source of the nitrate.

Water-Resources Investigations Report

Understanding contaminants associated with mineral deposits

Interdisciplinary studies by the U.S. Geological Survey (USGS) have resulted in substantial progress in understanding the processes that control the release of metals and acidic water from inactive mines and mineralized areas, the transport of metals and acidic water to streams, and the fate and effect of metals and acidity on downstream ecosystems. The potential environmental effects associated with abandoned and inactive mines, resulting from the complex interaction of a variety of chemical and physical processes, is an area of study that is important to the USGS Mineral Resources Program. Understanding the processes contributing to the environmental effects of abandoned and inactive mines is also of interest to a wide range of stakeholders, including both those responsible for managing lands with historically mined areas and those responsible for anticipating environmental consequences of future mining operations. The recently completed (2007) USGS project entitled 'Process Studies of Contaminants Associated with Mineral Deposits' focused on abandoned and inactive mines and mineralized areas in the Rocky Mountains of Montana, Colorado, New Mexico, Utah, and Arizona, where there are thousands of abandoned mines. Results from these studies provide new information that advances our understanding of the physical and biogeochemical processes causing the mobilization, transport, reaction, and fate of potentially toxic elements (including aluminum, arsenic, cadmium, copper, iron, lead, and zinc) in mineralized near-surface systems and their effects on aquatic and riparian habitat. These interdisciplinary studies provide the basis for scientific decisionmaking and remedial action by local, State, and Federal agencies charged with minimizing the effects of potentially toxic elements on the environment. Current (2007) USGS research highlights the need to understand (1) the geologic sources of metals and acidity and the geochemical reactions that release them from their sources, (2) the pathways that facilitate transport from those sources, and (3) the processes that control the fate of the elements once released from the sources. Experts in the fields of economic geology, structural geology, mineralogy, geophysics, geochemistry, hydrology, ground-water modeling, microbiology, and toxicology came together for a series of studies that address these relationships on scales ranging from the microscopic to the watershed. This Circular presents results and highlights from the detailed, interdisciplinary studies that include investigations in both mining-affected areas and mineralized but unmined areas. The first section of the Circular describes laboratory and site-scale field investigations that primarily focus on mineralogic and biologic controls on the source and release of metals and acidity from mine-waste rock and hydrothermally altered areas. The second section describes a set of basin- to watershed-scale studies that not only investigate the source and release of metals and acidity but also the transport of these constituents away from the source areas. The third section is a summary of results from postremediation ecosystem monitoring. For more information on these and other project-related studies, please visit the project Web site at http://minerals.cr.usgs.gov/projects/contaminants/index.html. The Web site includes a complete bibliography and detailed descriptions of each interdisciplinary study.

Circular

Herbicides and their transformation products in source-water aquifers tapped by public-supply wells in Illinois, 2001-02

During 2001-02, ground-water samples were collected from 117 public-supply wells distributed throughout Illinois to evaluate the occurrence of herbicides and their transformation products in the State?s source-water aquifers. Wells were selected using a stratified-random method to ensure representation of the major types of source-water aquifers in the State. Samples were analyzed for 18 herbicides and 18 transformation products, including 3 triazine and 14 chloroacetanilide products. Herbicide compounds (field-applied parent herbicides and their transformation products) were detected in 34 percent of samples. A subset of samples was collected unfiltered to determine if analytical results for herbicides in unfiltered samples are similar to those in paired filtered samples and, thus, can be considered equally representative of herbicide concentrations in ground water supplied to the public. The study by the U.S. Geological Survey was done in cooperation with the Illinois Environmental Protection Agency. Parent herbicides were detected in only 4 percent of all samples. The six most frequently detected herbicide compounds (from 5 to 28 percent of samples) were chloroacetanilide transformation products. The frequent occurrence of transformation products and their higher concentrations relative to those of most parent herbicides confirm the importance of obtaining information on transformation products to understand the mobility and fate of herbicides in ground-water systems. No sample concentrations determined during this study exceeded current (2003) Federal or State drinking-water standards; however, standards are established for only seven parent herbicides. Factors related to the occurrence of herbicide compounds in the State?s source-water aquifers include unconsolidated and unconfined conditions, various hydrogeologic characteristics and well-construction aspects at shallow depths, and proximity to streams. Generally, the closer an aquifer (or well location) is to a recharge area and (or) the stronger the hydraulic connection between an aquifer and a recharge area, the younger the ground water and the more vulnerable the aquifer will be to contamination by herbicide compounds. The weak relation between current (2001) statewide application rates of herbicides and current (2001-02) occurrence of herbicide compounds in source-water aquifers indicates that additional factors must be considered when relating herbicide-application rates to occurrence. These factors include historical application rates and the mobility and persistence of the various herbicide compounds in ground-water systems. Frequency of detection and concentrations of herbicides compounds in the State?s source-water aquifers are indicated to be highest during the spring, when crops are planted and herbicides primarily are applied. Excess nitrate (concentrations of nitrate, as nitrogen, higher than 3 milligrams per liter) in ground water strongly indicates the co-occurrence of herbicide compounds. However, nitrate concentrations are not a reliable indicator of herbicide-compound concentrations. The inverse relation found between current use of land for corn and soybean production and current occurrence of herbicide compounds in underlying aquifers indicates that various factors, along with current agricultural land use, contribute to herbicide occurrence. These factors include, among others, land-use history, ground-water age, ground-water-flow patterns, geology, soil microbiology, and chemistry and persistence of the herbicide compounds. Detection of agriculture-specific herbicide compounds in 71 percent of samples from urban areas with no current or recent agricultural land use near the sampled wells indicates that recharge to certain high-capacity supply wells may originate at considerable distances (up to about 10 miles) from the wells. Essentially no difference was found between the analytical results for herbicides in paired unfiltered and filtered samples,

Water-Resources Investigations Report

Ground-Water Quality of the Northern High Plains Aquifer, 1997, 2002-04

An assessment of ground-water quality in the northern High Plains aquifer was completed during 1997 and 2002-04. Ground-water samples were collected at 192 low-capacity, primarily domestic wells in four major hydrogeologic units of the northern High Plains aquifer-Ogallala Formation, Eastern Nebraska, Sand Hills, and Platte River Valley. Each well was sampled once, and water samples were analyzed for physical properties and concentrations of nitrogen and phosphorus compounds, pesticides and pesticide degradates, dissolved solids, major ions, trace elements, dissolved organic carbon (DOC), radon, and volatile organic compounds (VOCs). Tritium and microbiology were analyzed at selected sites. The results of this assessment were used to determine the current water-quality conditions in this subregion of the High Plains aquifer and to relate ground-water quality to natural and human factors affecting water quality. Water-quality analyses indicated that water samples rarely exceeded established U.S. Environmental Protection Agency public drinking-water standards for those constituents sampled; 13 of the constituents measured or analyzed exceeded their respective standards in at least one sample. The constituents that most often failed to meet drinking-water standards were dissolved solids (13 percent of samples exceeded the U.S. Environmental Protection Agency Secondary Drinking-Water Regulation) and arsenic (8 percent of samples exceeded the U.S. Environmental Protection Agency Maximum Contaminant Level). Nitrate, uranium, iron, and manganese concentrations were larger than drinking-water standards in 6 percent of the samples. Ground-water chemistry varied among hydrogeologic units. Wells sampled in the Platte River Valley and Eastern Nebraska units exceeded water-quality standards more often than the Ogallala Formation and Sand Hills units. Thirty-one percent of the samples collected in the Platte River Valley unit had nitrate concentrations greater than the standard, 22 percent exceeded the manganese standard, 19 percent exceeded the sulfate standard, 26 percent exceeded the uranium standard, and 38 percent exceeded the dissolved-solids standard. In addition, 78 percent of samples had at least one detectable pesticide and 22 percent of samples had at least one detectable VOC. In the Eastern Nebraska unit, 30 percent of the samples collected had dissolved-solids concentrations larger than the standard, 23 percent exceeded the iron standard, 13 percent exceeded the manganese standard, 10 percent exceeded the arsenic standard, 7 percent exceeded the sulfate standard, 7 percent exceeded the uranium standard, and 7 percent exceeded the selenium standard. No samples exceeded the nitrate standard. Thirty percent of samples had at least one detectable pesticide compound and 10 percent of samples had at least one detectable VOC. In contrast, the Sand Hills and Ogallala Formation units had fewer detections of anthropogenic compounds and drinking-water exceedances. In the Sand Hills unit, 15 percent of the samples exceeded the arsenic standard, 4 percent exceeded the nitrate standard, 4 percent exceeded the uranium standard, 4 percent exceeded the iron standard, and 4 percent exceeded the dissolved-solids standard. Fifteen percent of samples had at least one pesticide compound detected and 4 percent had at least one VOC detected. In the Ogallala Formation unit, 6 percent of water samples exceeded the arsenic standard, 4 percent exceeded the dissolved-solids standard, 3 percent exceeded the nitrate standard, 2 percent exceeded the manganese standard, 1 percent exceeded the iron standard, 1 percent exceeded the sulfate standard, and 1 percent exceeded the uranium standard. Eight percent of samples collected in the Ogallala Formation unit had at least one pesticide detected and 6 percent had at least one VOC detected. Differences in ground-water chemistry among the hydrogeologic units were attributed to variable depth to water, depth of the well screen below the water table, reduction-oxidation conditions, ground-water residence time, interactions with surface water, composition of aquifer sediments, extent of cropland, extent of irrigated land, and fertilizer application rates.

Scientific Investigations Report

Environmental factors and chemical and microbiological water-quality constituents related to the presence of enteric viruses in ground water from small public water supplies in southeastern Michigan

A study of small public ground-water-supply wells that produce water from discontinuous sand and gravel aquifers was done from July 1999 through July 2001 in southeastern Michigan. Samples were collected to determine the occurrence of viral pathogens and microbiological indicators of fecal contamination (“indicators”), determine whether indicators are adequate predictors of the presence of enteric viruses, and determine the factors that affect the presence of enteric viruses. Small systems are those that serve less than 3,300 people. Samples were analyzed for specific enteric viruses by reverse transcriptase-polymerase chain reaction (RT-PCR), for culturable viruses by cell culture, and for the indicators total coliforms, Escherichia coli (E. coli) , enterococci, and F-specific and somatic coliphage. Ancillary environmental and water-quality data were collected or compiled. A total of 169 regular samples and 32 replicate pairs were collected from 38 wells. Replicate pairs were samples collected at the same well on the same date. One well was sampled 6 times, 30 wells were sampled five times, 6 wells were sampled twice, and 1 well was sampled once. By use of RT-PCR, enterovirus was found in four wells (10.5 percent) and hepatitis A virus (HAV) in five wells (13.2 percent). In two of these wells, investigators found both enterovirus and HAV, but on different sampling dates. Culturable viruses were found one time in two wells (5.9 percent), and neither of these wells was positive for viruses by use of RT-PCR on any sampling date. If results for all viruses are combined, 9 of the 38 small public-supply wells were positive for enteric viruses (23.7 percent) by either cell culture or RT-PCR. One or more indicators were found in 18 of 38 wells. Total coliforms, E. coli , enterococci, and F-specific and somatic coliphage were found in 34.2, 10.5, 15.8, 5.9, and 5.9 percent, respectively, of the wells tested. In only 3 out of 18 wells were samples positive for an indicator on more than one date at the same well. The co-occurrence of enteric viruses and any indicator was 55.6 percent; five out of the nine virus-positive wells were also found to be positive for an indicator. Two wells with detections of viruses had a detection of total coliforms, one well had a detection of E. coli , one of enterococci, and one of F-specific coliphage. On a per sample basis, of 11 samples that were positive for enteric viruses, indicator bacteria co-occurred in only 2 samples, and coliphage were not present in any. More virus-positive samples were found at sites served by septic systems than those served by sewerlines. Sampling condition (ground water or a mixture of tank and ground water), distance to septic system, type of and distance to nearest surface-water body, well characteristics, or land use were not related to the presence of viruses or indicators. Among continuous water-quality variables, statistically significant relations were found between total coliforms and dissolved organic carbon and between total coliforms and iron. There was a statistically significant relation between chloride concentrations >20 mg/L and detections of total coliforms. Presence of nitrate and nitrite was related to the presence of other indicators ( E. coli , enterococci, and F-specific and somatic coliphage) or enteric viruses, but not to total coliforms. The data indicated that chloride-to-bromide (C1:Br) ratios may be useful as a screening tool for total coliforms and enteric viruses but not for E. coli , enterococci, and F-specific and somatic coliphage. This study provides evidence for fecal contamination of ground water from small public-supply wells, at least on an intermittent basis. Collecting data on multiple lines of evidence would be needed to reliably predict the presence of enteric viruses and protect public health. Future data collection toward this end could include repeat sampling several times a year for different indicators, measuring dissolved-organic carbon, nitrate plus nitrite, and (or) chloride concentrations, or determining C1:Br ratios. The presence of a site served by a septic system is an indication that the well may be more vulnerable to contamination than a site served by a sewerline.

Michigan

Water Quality in the Mahoning River and Selected Tributaries in Youngstown, Ohio

The lower reaches of the Mahoning River in Youngstown, Ohio, have been characterized by the Ohio Environmental Protection Agency (OEPA) as historically having poor water quality. Most wastewater-treatment plants (WWTPs) in the watershed did not provide secondary sewage treatment until the late 1980s. By the late 1990s, the Mahoning River still received sewer-overflow discharges from 101 locations within the city of Youngstown, Ohio. The Mahoning River in Youngstown and Mill Creek, a principal tributary to the Mahoning River in Youngstown, have not met biotic index criteria since the earliest published assessment by OEPA in 1980. Youngstown and the OEPA are working together toward the goal of meeting water-quality standards in the Mahoning River. The U.S. Geological Survey collected information to help both parties assess water quality in the area of Youngstown and to estimate bacteria and inorganic nitrogen contributions from sewer-overflow discharges to the Mahoning River. Two monitoring networks were established in the lower Mahoning River: the first to evaluate hydrology and microbiological and chemical water quality and the second to assess indices of fish and aquatic-macroinvertebrate-community health. Water samples and water-quality data were collected from May through October 1999 and 2000 to evaluate where, when, and for how long water quality was affected by sewer-overflow discharges. Water samples were collected during dry- and wet-weather flow, and biotic indices were assessed during the first year (1999). The second year of sample collection (2000) was directed toward evaluating changes in water quality during wet-weather flow, and specifically toward assessing the effect of sewer-overflow discharges on water quality in the monitoring network. Water-quality standards for Escherichia coli ( E. coli ) concentration and draft criteria for nitrate plus nitrite and total phosphorus were the regulations most commonly exceeded in the Mahoning River and Mill Creek sampling networks. E. coli concentrations increased during wet-weather flow and remained higher than dry-weather concentrations for 48 hours after peak flow. E. coli concentration criteria were more commonly exceeded during wet-weather flow than during dry-weather flow. Exceedances of nutrient-concentration criteria were not substantially more common during wet-weather flow. The fish and aquatic macroinvertebrate network included Mill Creek and its tributaries but did not include the main stem of the Mahoning River. Persistent exceedances of chemical water-quality standards in Mill Creek and the presence of nutrient concentrations in excess of draft criteria may have contributed to biotic index scores that on only one occasion met State criteria throughout the fish and aquatic macroinvertebrate sampling network. Monitored tributary streams did not contribute concentrations of E. coli , nitrate plus nitrite, or total phosphorus to the Mahoning River and Mill Creek that were higher than main-stem concentrations, but monitored WWTP and sewer-overflow discharges did contribute. Twenty-four hour load estimates of sewer-overflow discharge contributions during wet-weather flow indicated that sewer-overflow discharges contributed large loads of bacteria and inorganic nitrogen to the Mahoning River relative to the instream load. The sewer-overflow loads appeared to move as a slug of highly enriched water that passed through Youngstown on the rising limb of the storm hydrograph. The median estimated sewer-overflow load contribution of bacteria was greater than the estimated instream load by a factor of five or more; however, the median estimated sewer-overflow load of inorganic nitrogen was less than half of the estimated instream load. Sewer-overflow discharges contributed loads of E. coli and nutrients to the Mahoning River and Mill Creek at a point where the streams already did not meet State water-quality regulations. Improvement of water quality of the Mahoning River, Mill Creek, and tributaries at Youngstown would be facilitated by reducing loads from sewer-overflow discharges within Youngstown, by identifying and reducing other sources of E. coli and nutrients within Young-stown, and by reducing discharges of E. coli , nitrate plus nitrite, and total phosphorus to the Mahoning River and Mill Creek upstream from Youngstown.

Water-Resources Investigations Report

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