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A supplement to "Methods for collection and analysis of aquatic biological and microbiological samples"

The report contains methods used by the U.S. Geological Survey to collect, preserve, and analyze waters to determine their biological and microbiological properties. It supplements, "Methods for Collection and Analysis of Aquatic Biological and Microbiological Samples" (TWRI, Book 5, Chapter A4, 1977, edited by P. E. Greeson, T. A. Ehlke, G. A. Irwin, B. W. Lium, and K. V. Slack). Included in the supplement are 5 new methods, a new section of selected taxonomic references for Ostracoda, and 6 revised methods.

Techniques of Water-Resources Investigations↗

Microbiological oxidation of antimony(III) with oxygen or nitrate by bacteria isolated from contaminated mine sediments

Bacterial oxidation of arsenite [As(III)] is a well-studied and important biogeochemical pathway that directly influences the mobility and toxicity of arsenic in the environment. In contrast, little is known about microbiological oxidation of the chemically similar anion antimonite [Sb(III)]. In this study, two bacterial strains, designated IDSBO-1 and IDSBO-4, which grow on tartrate compounds and oxidize Sb(III) using either oxygen or nitrate, respectively, as a terminal electron acceptor, were isolated from contaminated mine sediments. Both isolates belonged to the Comamonadaceae family and were 99% similar to previously described species. We identify these novel strains as Hydrogenophaga taeniospiralis strain IDSBO-1 and Variovorax paradoxus strain IDSBO-4. Both strains possess a gene with homology to the aioA gene, which encodes an As(III)-oxidase, and both oxidize As(III) aerobically, but only IDSBO-4 oxidized Sb(III) in the presence of air, while strain IDSBO-1 could achieve this via nitrate respiration. Our results suggest that expression of aioA is not induced by Sb(III) but may be involved in Sb(III) oxidation along with an Sb(III)-specific pathway. Phylogenetic analysis of proteins encoded by the aioA genes revealed a close sequence similarity (90%) among the two isolates and other known As(III)-oxidizing bacteria, particularly Acidovorax sp. strain NO1. Both isolates were capable of chemolithoautotrophic growth using As(III) as a primary electron donor, and strain IDSBO-4 exhibited incorporation of radiolabeled [ 14 C]bicarbonate while oxidizing Sb(III) from Sb(III)-tartrate, suggesting possible Sb(III)-dependent autotrophy. Enrichment cultures produced the Sb(V) oxide mineral mopungite and lesser amounts of Sb(III)-bearing senarmontite as precipitates.

Idaho↗

Using chemical and microbiological indicators to track the impacts from the land application of treated municipal wastewater and other sources on groundwater quality in a karstic springs basin

Multiple chemical constituents (nutrients; N, O, H, C stable isotopes; 64 organic wastewater compounds, 16 pharmaceutical compounds) and microbiological indicators were used to assess the impact on groundwater quality from the land application of approximately 9.5 million liters per day of treated municipal sewage effluent to a sprayfield in the 960-km2 Ichetucknee Springs basin, northern Florida. Enriched stable isotope signatures (?? 18O and ??2H) were found in water from the effluent reservoir and a sprayfield monitoring well (MW-7) due to evaporation; however, groundwater samples downgradient from the sprayfield have ??18O and ??2H concentrations that represented recharge of meteoric water. Boron and chloride concentrations also were elevated in water from the sprayfield effluent reservoir and MW-7, but concentrations in groundwater decreased substantially with distance downgradient to background levels in the springs (about 12 km) and indicated at least a tenfold dilution factor. Nitrate-nitrogen isotope (??15N-NO3) values above 10 ??? in most water samples were indicative of organic nitrogen sources except Blue Hole Spring (??15N-NO3 = 4.6-4.9 ???), which indicated an inorganic source of nitrogen (fertilizers). The detection of low concentrations the insect repellent N,N-diethyl-metatoluamide (DEET), and other organic compounds associated with domestic wastewater in Devil's Eye Spring indicated that leakage from a nearby septic tank drainfield likely has occurred. Elevated levels of fecal coliforms and enterococci were found in Blue Hole Spring during higher flow conditions, which likely resulted from hydraulic connections to upgradient sinkholes and are consistent with previoius dye-trace studies. Enteroviruses were not detected in the sprayfield effluent reservoir, but were found in low concentrations in water samples from a downgradient well and Blue Hole Spring during high-flow conditions indicating a human wastewater source. The Upper Floridan aquifer in the Ichetucknee Springs basin is highly vulnerable to contamination from multiple anthropogenic sources throughout the springs basin. ?? 2007 Springer-Verlag.

Environmental Geology↗

Organic and inorganic composition and microbiology of produced waters from Pennsylvania shale gas wells

Hydraulically fractured shales are becoming an increasingly important source of natural gas production in the United States. This process has been known to create up to 420 gallons of produced water (PW) per day, but the volume varies depending on the formation, and the characteristics of individual hydraulic fracture. PW from hydraulic fracturing of shales are comprised of injected fracturing fluids and natural formation waters in proportions that change over time. Across the state of Pennsylvania, shale gas production is booming; therefore, it is important to assess the variability in PW chemistry and microbiology across this geographical span. We quantified the inorganic and organic chemical composition and microbial communities in PW samples from 13 shale gas wells in north central Pennsylvania. Microbial abundance was generally low (66–9400 cells/mL). Non-volatile dissolved organic carbon (NVDOC) was high (7–31 mg/L) relative to typical shallow groundwater, and the presence of organic acid anions (e.g., acetate, formate, and pyruvate) indicated microbial activity. Volatile organic compounds (VOCs) were detected in four samples (∼1 to 11.7 μg/L): benzene and toluene in the Burket sample, toluene in two Marcellus samples, and tetrachloroethylene (PCE) in one Marcellus sample. VOCs can be either naturally occurring or from industrial activity, making the source of VOCs unclear. Despite the addition of biocides during hydraulic fracturing, H 2 S-producing, fermenting, and methanogenic bacteria were cultured from PW samples. The presence of culturable bacteria was not associated with salinity or location; although organic compound concentrations and time in production were correlated with microbial activity. Interestingly, we found that unlike the inorganic chemistry, PW organic chemistry and microbial viability were highly variable across the 13 wells sampled, which can have important implications for the reuse and handling of these fluids

Pennsylvania↗

Strong nonlinear photonic responses from microbiologically synthesized tellurium nanocomposites

A new class of nanomaterials, namely microbiologically-formed nanorods composed of elemental tellurium [Te(0)] that forms unusual nanocomposites when combined with poly(m-phenylenevinylene-co-2,5-dioctoxy-phenylenevinylene) (PmPV) is described. These bio-nanocomposites exhibit excellent broadband optical limiting at 532 and 1064 nm. Nonlinear scattering, originating from the laser induced solvent bubbles and microplasmas, is responsible for this nonlinear behavior. The use of bacterially-formed Te(0) when combined with an organic chemical host (e.g., PmPV) is a new green method of nanoparticle syntheses. This opens the possibilities of using unique, biologically synthesized materials to advance future nanoelectronic and nanophotonic applications.

Chemical Physics Letters↗

Perspectives on the future of host-microbe biology from the Council on Microbial Sciences of the American Society for Microbiology

Host-microbe biology (HMB) stands on the cusp of redefinition, challenging conventional paradigms to instead embrace a more holistic understanding of the microbial sciences. The American Society for Microbiology (ASM) Council on Microbial Sciences hosted a virtual retreat in 2023 to identify the future of the HMB field and innovations needed to advance the microbial sciences. The retreat presentations and discussions collectively emphasized the interconnectedness of microbes and their profound influence on humans, animals, and environmental health, as well as the need to broaden perspectives to fully embrace the complexity of these interactions. To advance HMB research, microbial scientists would benefit from enhancing interdisciplinary and transdisciplinary research to utilize expertise in diverse fields, integrate different disciplines, and promote equity and accessibility within HMB. Data integration will be pivotal in shaping the future of HMB research by bringing together varied scientific perspectives, new and innovative techniques, and ’omics approaches. ASM can empower under-resourced groups with the goal of ensuring that the benefits of cutting-edge research reach every corner of the scientific community. Thus, ASM will be poised to steer HMB toward a future that champions inclusivity, innovation, and accessible scientific progress.

mSphere↗

Contamination assessment in microbiological sampling of the Eyreville core, Chesapeake Bay impact structure

Knowledge of the deep subsurface biosphere is limited due to difficulties in recovering materials. Deep drilling projects provide access to the subsurface; however, contamination introduced during drilling poses a major obstacle in obtaining clean samples. To monitor contamination during the 2005 International Continental Scientific Drilling Program (ICDP)-U.S. Geological Survey (USGS) deep drilling of the Chesapeake Bay impact structure, four methods were utilized. Fluorescent microspheres were used to mimic the ability of contaminant cells to enter samples through fractures in the core material during retrieval. Drilling mud was infused with a chemical tracer (Halon 1211) in order to monitor penetration of mud into cores. Pore water from samples was examined using excitation-emission matrix (EEM) fl uorescence spectroscopy to characterize dissolved organic carbon (DOC) present at various depths. DOC signatures at depth were compared to signatures from drilling mud in order to identify potential contamination. Finally, microbial contaminants present in drilling mud were identified through 16S ribosomal deoxyribonucleic acid (rDNA) clone libraries and compared to species cultured from core samples. Together, these methods allowed us to categorize the recovered core samples according to the likelihood of contamination. Twenty-two of the 47 subcores that were retrieved were free of contamination by all the methods used and were subsequently used for microbiological culture and culture-independent analysis. Our approach provides a comprehensive assessment of both particulate and dissolved contaminants that could be applied to any environment with low biomass. ?? 2009 The Geological Society of America.

Special Paper of the Geological Society of America↗

Biological and microbiological assessment of the upper Chattahoochee River basin, Georgia

Biological and microbiological studies were conducted by the U.S. Geological Survey as a part of the Intensive River-Quality Assessment studies of the upper Chattahoochee River basin, Georgia. Phytoplankton concentrations in cells per milliliter (cells/mL) were generally higher downstream from Atlanta than upstream. The highest concentrations, mostly blue-green algae, occurred in West Point Lake with an average of 90,000 cells/mL for the sampling period. The lowest concentrations, 1,000 cells/mL, occurred upstream of Lake Sidney Lanier. Dissolved orthophosphate and nitrite plus nitrate concentrations were highest in the river reaches and upper reaches of the two lakes and were lowest at the dam pools of both lakes. The high nitrite plus nitrate concentrations downstream from Atlanta were primarily a result of nitrification by Nitrosomonas and Nitrobacter bacteria. Algal growth potential was highest downstream from Atlanta, 25 milligrams per liter (mg/L) at Whitesburg, and was the lowest in the headwaters and at the dam pools of Lake Sidney Lanier and West Point Lake. The rate of nitrification in the Atlanta to Franklin reach of the river was comparatively low, 0.02 mg/L per hour. Nitrification was an important cause of dissolved-oxygen consumption in a 45-mi reach of the river downstream from the Atlanta wastewater treatment facilities. Dissolved-oxygen consumption as a result of nitrification may be greatest during low flow. (Woodard-USGS)

Alabama, Georgia↗

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↗

Chemical and microbiological monitoring of a sole-source aquifer intended for artificial recharge, Nassau County, New York

In late 1980, approximately 4 million gallons per day of highly treated wastewater will be used to recharge the groundwater reservoir in central Nassau County through a system of 10 recharge basins and 5 shallow injection wells. To evaluate the impact of large-scale recharge with reclaimed water on groundwater quality, the U.S. Geological Survey has collected hydrologic and water-quality data from a 1-square-mile area around the recharge site to provide a basis for future comparison. Extensive chemical and microbiological analyses are being made on samples from 48 wells screened in the upper glacial (water-table) aquifer and the upper part of the underlying Magothy (public-supply) aquifer. Preliminary results indicate that water from the upper glacial aquifer contains significant concentrations of nitrate and low-molecular-weight chlorinated hydrocarbons and detectable concentrations of organochlorine insecticides and polychlorinated biphenyls. At present, no fecal contamination is evident in either aquifer in the area studied. In the few samples containing fecal indicator bacteria, the numbers were low. Nonpoint sources provide significant loads of organic and inorganic compounds; major sources include cesspool and septic-tank effluent, cesspool and septic-tank cleaners and other over-the-counter domestic organic solvents, fertilizers, insecticides for termite and other pest control, and stormwater runoff to recharge basins. The water-table aquifer is composed mainly of stratified, well-sorted sand and gravel and, as a result, is highly permeable. In the 1-square-mile area studied, some contaminants seem to have traveled 200 feet downward to the bottom of the water-table aquifer and into the upper part of the public-supply aquifer. (USGS)

Open-File 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↗

Assessment of possible sources of microbiological contamination in the water column and streambed sediment of the Jacks Fork, Ozark National Scenic Riverways, Missouri — Phase III

In 1998, a 5 river-mile reach of the Jacks Fork was included on Missouri's list of impaired waters as required by Section 303(d) of the Federal Clean Water Act. The identified pollutant on the Jacks Fork was fecal coliform bacteria. The length of the impaired reach was changed to 7 miles on the Missouri 2002 303(d) list because of data indicating the fecal coliform bacteria problem existed over a broader area. The U.S. Geological Survey, in cooperation with the National Park Service, conducted a study to better understand the extent and sources of microbiological contamination within the Jacks Fork from Alley Spring to the mouth, which includes the 7-mile 303(d) reach. Ten sites were sampled from June 2003 through October 2003 and from June 2004 through October 2004. Water-column and streambed sediment samples were collected from main-stem and tributary sites mostly during base-flow conditions during a variety of recreational season river uses and analyzed for fecal coliform and Escherichia coli bacteria. Isolates of Escherichia coli obtained from water samples collected at five sites were submitted for rep-PCR analysis to identify presumptive sources of fecal indicator bacteria in the Jacks Fork. Results indicate that recreational users (including boaters and swimmers) are not the primary source of fecal coliform bacteria in the Jacks Fork; rather, the presence of fecal coliform bacteria is associated with other animals, of which horses are the primary source. Increases in fecal coliform bacteria densities in the Jacks Fork are associated with cross-country horseback trail-riding events.

Missouri↗

Compositional analysis of formation water geochemistry and microbiology of commercial and carbon dioxide-rich wells in the southwestern United States

Studies of naturally occurring subsurface carbon dioxide (CO 2 ) accumulations can provide useful information for potential CO 2 injection projects; however, the microbial communities and formation water geochemistry of most reservoirs are understudied. Formation water and microbial biomass were sampled at four CO 2 -rich reservoir sites: two within Bravo Dome, a commercial CO 2 field in New Mexico; one northwest of Bravo Dome in Colorado (Oakdale Field); and one southwest of Bravo Dome in New Mexico (Rafter “K” Ranch). Aside from the Rafter “K” Ranch site, minor differences were observed in the geochemistry of formation water collected from these sites compared to historical data. No organisms were significantly associated with Oakdale Field compared to the other three sites, nor were any hydrogeochemical or gas geochemical parameters (for example, CO 2 concentration) found to have significant associations with the microbial ecology of these four sites. Microorganisms from these sites were metabolically diverse and had the potential to (1) generate methane, (2) produce corrosive hydrogen sulfide (H 2 S), and (3) rapidly biofoul and (or) clog pore spaces by shifting microbial communities with changes in salinity or nutrient supply. This study demonstrates that high concentrations of CO 2 in subsurface reservoirs apparently have not imparted a distinct geochemical or microbiological signature on the associated formation waters and that the microorganisms in these reservoirs are metabolically diverse and could adapt to geochemical changes in the subsurface.

Colorado, New Mexico, Texas, Oklahoma↗

Methods for collection and analysis of aquatic biological and microbiological samples

Chapter A4 contains methods used by the U.S. Geological Survey to collect, preserve, and analyze waters to determine their biological and microbiological properties. Part 1 discusses biological sampling and sampling statistics. The statistical procedures are accompanied by examples. Part 2 consists of detailed descriptions of more than 45 individual methods, including those for bacteria, phytoplankton, zooplankton, seston, periphyton, macrophytes, benthic invertebrates, fish and other vertebrates, cellular contents, productivity, and bioassays. Each method is summarized, and the application, interferences, apparatus, reagents, collection, analysis, calculations, reporting of results, precision and references are given. Part 3 consists of a glossary. Part 4 is a list of taxonomic references.

Techniques of Water-Resources Investigations↗

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, orthophosphate, calcium, magnesium, sodium, potassium, 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 polymerase chain-reaction analysis. Twenty-nine percent of the samples (26 of 90) had bacterial contamination. 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 samples (3 of 87) had oxidized nitrogen concentrations 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 correlation with the concentration of sulfate and sample pH as measured at the U.S. Geological Survey National Water-Quality Laboratory in Denver, Colorado.

Maryland↗

Microbiological quality of water from noncommunity supply wells in carbonate and crystalline aquifers of Pennsylvania

Samples were collected from 59 noncommunity water supplies in the Commonwealth of Pennsylvania from September 2000 to January 2001 and analyzed for pathogens and microbiological indicator organisms. The pathogens sampled were culturable viruses and Helicobacter pylori (H. pylori). The indicator organisms sampled were total coliform, Escherichia coli ( E. coli ), Clostridium perfringens ( C. perfringens ), somatic coliphage, male- specific coliphage, and enterococcus. The two primary areas sampled for the project completed by the U.S. Geological Survey, in cooperation with the Pennsylvania Department of Environmental Protection (PaDEP), were carbonate aquifers and crystalline aquifers. The results of all sampling showed culturable viruses were detected in 8 percent of the wells, H. pyloriin 7 percent of the wells, E. coli in 12 percent of the wells, total coliform in 46 percent of the wells, C. perfringens in 15 percent of the wells, somatic coliphage in 8 percent of the wells, male-specific coliphage in 5 percent of the wells, and enterococcus in 14 percent of the wells. Carbonate aquifers tended to have higher detection rates for the pathogens and indicators sampled than the crystalline aquifers. Detections of the pathogens and indicator organisms were not related statistically to the amounts of urban, agricultural, or forested area in a 1,500-foot radius around the sampled well. Somatic and male-specific coliphage showed the best relation to occurrence of culturable viruses. Culturable viruses and H. pylori were detected in wells in which no indicator organisms were present; therefore, none of the indicator organisms sampled provide complete assurance of pathogenfree water. The best predictive tool for virus screening was a combination of indicator organisms.

Water-Resources Investigations Report↗

Assessment of possible sources of microbiological contamination and water-quality characteristics of the Jacks Fork, Ozark National Scenic Riverways, Missouri — Phase II

In 1998, an 8-mile reach of the Jacks Fork was included on Missouri's list of impaired waters as required by Section 303(d) of the Federal Clean Water Act. The identified pollutant on the Jacks Fork was fecal coliform bacteria. Potential sources of fecal contamination to the Jacks Fork include a wastewater treatment plant; campground pit-toilet or septic-system effluent; a large commercial, cross-country horseback trail riding facility; canoeists, boaters, and tubers; and cows. The U.S. Geological Survey, in cooperation with the National Park Service, conducted a study to better understand the extent and sources of microbiological contamination within the Jacks Fork from Alley Spring to the mouth, which includes the 8-mile 303(d) reach. Identification of the sources would provide the National Park Service and the State of Missouri with the information needed to craft a solution of abatement, regulation, prevention, and mitigation with the end result being the removal of the Jacks Fork from the 303(d) list. Fifteen sites were sampled from November 1999 through December 2000. An additional site was sampled one time. Samples were collected mostly during base-flow conditions during a variety of nonrecreational and recreational season river uses. Samples were analyzed for selected fecal indicator bacteria, physical properties, nutrients, and wastewater organic compounds. During the sampling period, the whole-body-contact recreation standard for fecal coliform (200 colonies per 100 milliliters of sample) was exceeded at three sites on August 10, 2000, and also at one site on May 11, June 7, and October 3, 2000. Fecal coliform densities and instantaneous loads generally increased from background concentrations at the Eminence site, peaked about 2 river miles downstream, and then decreased until the most downstream site sampled. Generally, the largest densities and loads at sites downstream from Eminence not related to wet-weather flow were observed during a trail ride held August 6 to 12, 2000. A 24-hour sample collection effort was conducted the weekend of July 15 and 16, 2000, to investigate the effect that large numbers of swimmers, canoeists, and tubers had on fecal coliform densities in the Jacks Fork. Five or six samples were collected at six sites between Saturday morning and the following Sunday afternoon. No fecal coliform density at any of the sites sampled exceeded the whole-body-contact recreation standard. Because bacteria survive longer in stream-bed sediments than in water, a source of bacteria in the water column could be from resuspension of accumulated bacteria from streambed sediments. Water and streambed-sediment samples were collected at three sites on August 3, 2000, 1 week before a trail ride and again at three sites on August 8, 2000, during a trail ride. Sixty-five Escherichia coli isolates obtained from water samples collected at 9 sites and 23 Escherichia coli isolates obtained from stream-bed-sediment samples collected at 5 sites were submitted for ribotyping analysis. Samples were collected in 2000 during a variety of nonrecreational and recreational season river uses, including trail rides, canoeing, tubing, and swimming. Of the 65 isolates from water samples, 40 percent were identified as originating from sewage, 29 percent from horse, 11 percent from cow, and 20 percent from an unknown source. Of the 23 isolates from streambed-sediment samples, 39 percent were identified as originating from sewage, 35 percent from horse, 13 percent from cow, and 13 percent from unknown sources. Analysis of physical property (dissolved oxygen, pH, specific conductance, and temperature) and nutrient (dissolved nitrite plus nitrate and total phosphorus) data indicated that overall few statistically significant differences occurred among the main stem sites of the Jacks Fork. A significant increase in total phosphorus concentrations did occur at site 75 immediately downstream from the Eminence Wastewater Treatment Plant, but the effect diminished quickly downstream. Unlike fecal coliform bacteria, most variations in physical property values or nutrient concentrations were related to seasonal changes, time of day the sample was collected, or hydrologic conditions and not to certain recreational activities. Trace quantities of wastewater organic compounds were detected in all waters sampled for these constituents. Two of the compounds were detected in associated laboratory blanks, and other detected compounds have sources other than sewage effluent. The best indicators of municipal or domestic sewage effluent were the non-ionic detergent metabolites (nonylphenol monoethoxylate, octylphenol monoethoxylate, and para-nonylphenol), phenol, and caffeine; but possible sources of these compounds, which were detected in one or more of the samples, could be the numerous campers, swimmers, and canoeists that were present when the samples were collected.

Missouri↗

Microbiological and chemical quality of ground water used as a source of public supply in southern Missouri — Phase II, April–July, 1998

The protection of public health through quality public ground-water systems is the responsibility of the U. S. Environmental Protection Agency and the State of Missouri, through the Missouri Department of Natural Resources, Public Drinking Water Program. Approximately 95 percent of the public-water supplies in Missouri use ground water as their source of drinking water through more than 3,700 public wells. Karst terrain, intensive agricultural operations, extensive numbers of on-site sewage systems, and poor well construction can lead to chemical and microbiological contamination of the contributing aquifers. Sitespecific studies and routine regulatory monitoring have produced information on the overall quality and potability of the State's public-drinking-water supplies, but little is known about the presence of viruses. The U.S. Geological Survey, in cooperation with the Missouri Department of Natural Resources, sampled 109 public-water supplies to characterize the physical, chemical, bacterial, and viral conditions in southern Missouri. During April to July 1998, these wells were sampled for nutrients, total organic carbon, optical brighteners, indicator bacteria, enteric viruses, and ribonucleic acid and somatic coli phages. These constituents indicate possible surface contamination of the sampled aquifer. Selection of the wells to be sampled depended on the age of the well (pre-1970), land use, geohydrology, and well construction. None of the physical or chemical constituents measured or analyzed exceeded Missouri's Drinking Water Standards set by the Public Drinking Water Program of the Missouri Department of Natural Resources. The majority of ammonia plus organic nitrogen, nitrite, and phosphorus concentrations were below the laboratory's minimum reporting levels. There were a greater number of detects above the minimum reporting level with respect to the nitrite plus nitrate, ammonia, orthophosphate, and total organic carbon concentrations. Analyses included comparing and contrasting the data by grouping according to well age and construction, karst type, geohydrology, soil type, and land use. There was little variation in well construction between selected wells. The results indicated several groupings of similar and dissimilar concentrations, most expected because of hydrological, physical, or land use differences. Dissolved oxygen values indicated distinct variation in the different groupings. There were significant differences in dissolved oxygen values between the secondary and non-karst areas, the Ozark confined and Ozark unconfined geohydrologic groups, and between agricultural and other land uses. In groupings by soil and geohydrology, the Missouri bootheel region differed with respect to ammonia, total organic carbon, and phosphorus when compared with the other groups. Less than 10 percent of the wells sampled tested positive for bacterial contamination. E. coli was the most frequently detected bacterium. The public wells at Monett and West Plains, Missouri, had plates with colonies too numerous to count for all three indicator bacteria. Further analyses by rRNA (ribosomal RiboNucleic Acid) hybridization techniques detennined that much of the bacteria present were from ruminant and human sources. No enteric viruses were detected in the 109 samples. Both ribonucleic acid and somatic coliphage were detected at two wells. One additional well had ribonucleic acid coliphage and another had somatic coliphage for a total of four wells with coliphage selects.

Missouri↗