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Proceedings of the U.S. Geological Survey Interdisciplinary Microbiology Workshop, Estes Park, Colorado, October 15-17, 2008

Preface A U.S. Geological Survey Interdisciplinary Microbiology Workshop was held in Estes Park, Colorado, on October 15-17, 2008. Participants came from all USGS regions and disciplines. This report contains abstracts from 36 presentations and 35 poster sessions and notes from 5 breakout sessions. The seven presentation topics follow: Ecology of wildlife and fish disease Mechanisms of fish and wildlife disease Microbial ecology Geographic patterns/visualization Public health and water quality Geomicrobiology Ecosystem function The six poster session topics follow: Wildlife disease Disease detection methods Water quality Microbial ecology Metabolic processes Tools and techniques Five working groups met in breakout sessions on October 16, 2008. The highlights for each working group are summarized in this report, and their goals are listed below: Working Group I: to plan a Fact Sheet on interdisciplinary microbiology in the USGS Working Group II: to plan a USGS interdisciplinary microbiology Web site Working Group III: to suggest ways to broadcast and publicize the types of microbiology conducted at the USGS Working Group IV: to identify emerging issues in USGS interdisciplinary microbiology research Working Group V: to identify potential opportunities for interdisciplinary microbiology work at the USGS After the workshop, the USGS interdisciplinary microbiology Web site was activated in June 2009 at http://microbiology.usgs.gov/.

Scientific Investigations Report

Microbiological monitoring for the U.S. Geological Survey National Water-Quality Assessment Program

Data to characterize the microbiological quality of the Nation's fresh, marine, and estuarine waters are usually collected for local purposes, most often to judge compliance with standards for protection of public health in swimmable or drinkable waters. Methods and procedures vary with the objectives and practices of the parties collecting data and are continuously being developed or modified. Therefore, it is difficult to provide a nationally consistent picture of the microbial quality of the Nation's waters. Study objectives and guidelines for a national microbiological monitoring program are outlined in this report, using the framework of the U.S. Geological Survey (USGS) National Water-Quality Assessment (NAWQA) program. A national program is designed to provide long-term data on the presence of microbiological pathogens and indicators in ground water and surface water to support effective water policy and management. Three major groups of waterborne pathogens affect the public health acceptability of waters in the United States—bacteria, protozoa, and viruses. Microbiological monitoring in NAWQA would be designed to assess the occurrence, distribution, and trends of pathogenic organisms and indicators in surface waters and ground waters; relate the patterns discerned to factors that help explain them; and improve our understanding of the processes that control microbiological water quality.

Water-Resources Investigations Report

Perspectives on the future of ecology, evolution, and biodiversity from the Council on Microbial Sciences of the American Society for Microbiology

The field of microbial ecology, evolution, and biodiversity (EEB) is at the leading edge of understanding how microbes shape our biosphere and influence the well-being of humankind. To that end, EEB is developing new tools to analyze the massive, complex, transdisciplinary datasets that result from such studies. The American Society for Microbiology’s Council on Microbial Sciences hosted a virtual retreat in 2023 to identify the trajectory of EEB both within the Society and microbiology at large. Retreat presentations and discussions collectively emphasized the interconnectedness of microbes and their outsized global influence on environmental and host health. The retreat repeatedly emphasized that the future of science is through transdisciplinary collaborations that promote integration of diverse datasets and advanced technologies, while also genuinely fostering inclusivity, global collaborations, and public engagement. The American Society for Microbiology and EEB stand poised to lead a paradigm shift that will result in a new era of collaboration, innovation, and societal relevance for microbiology.

mSphere

Geochemistry and microbiology of groundwater and solids from extraction and monitoring wells and their relation to well efficiency at a Federally operated confined disposal facility, East Chicago, Indiana

In cooperation with the U.S. Army Corps of Engineers, Chicago District, the U.S. Geological Survey investigated the processes affecting water quality, geochemistry, and microbiology in representative extraction and monitoring wells at a confined disposal facility (CDF) in East Chicago, Indiana. The CDF is a 140-acre Federally-managed facility that was the former location of an oil refinery and is now used for the long-term disposal and storage of dredge material from the Indiana Harbor and Indiana Harbor Canal. Residual petroleum hydrocarbons and leachate from the CDF are contained within the facility by use of a groundwater cutoff wall. The wall consists of a soil-bentonite slurry and a gradient control system made up of an automated network of 96 extraction wells, 42 monitoring wells, and 2 ultrasonic sensors that maintain an inward hydraulic gradient at the site. The pumps in the extraction wells require vigilant maintenance and must be replaced when unable to withdraw water at a rate sufficient to maintain the required inward gradient. The wells are screened in the Calumet aquifer, a coarse-grained sand and gravel unit that extends approximately 35 feet below the land surface and is not utilized for drinking-water supply at the CDF or in the surrounding area. This study was initiated to identify the cause of decreased pump discharges and to identify potential mitigation strategies. For this study, the U.S. Geological Survey collected groundwater and solids from monitoring and extraction wells. Groundwater samples were collected during June 2014 for precautionary health screening and on four occasions during September 2014 through November 2014. Groundwater samples collected from two extraction wells during June 2014 were analyzed for concentrations of anthropogenic organic constituents. During September through November 2014, groundwater samples were collected from one additional extraction well, and samples from three monitoring wells were analyzed for concentrations of inorganic and organic constituents, dissolved gases, and bacterial abundance and diversity. Solid samples were collected during April 2014, during September 2014 through November 2014, and during November 2016. Solid samples were collected from the exterior of extraction-well pumps and as flocculent from water samples. Solid samples were collected from 10 wells, including 1 extraction well and 3 monitoring wells sampled for water quality. Solid samples were analyzed for mineralogy, solid-phase habit, geochemistry, and organic composition. The following is a list of observations that were made during this study: (1) the water quality is substantially variable among the six well locations sampled as part of this study—lower (more negative) redox values and higher concentrations of many constituents (including calcium, magnesium, sodium, and sulfate) and properties (including dissolved solids, hardness, and turbidity) were detected in sampled wells located near the extraction wells with the highest frequency of failure; (2) water-level drawdown is variable between extraction wells—wells with the greatest drawdown may pull deeper groundwater into the borehole; (3) dissolved gas results indicate reducing oxidation-reduction processes in the aquifer material that can feasibly contribute iron, carbon dioxide, and other byproducts from hydrocarbon degradation to precipitates and solids that accumulate on and impair pump operation; (4) crystalline and amorphous solid-phase minerals are precipitating in the borehole; (5) several types of bacteria are present in water pumped from extraction wells and are likely responsible for bonding mineral and microbiologic matter to the pump (and other well components); and (6) bacteria may create microenvironments that facilitate precipitation of solids or inhibit dissolution of unstable minerals once the bacteria adhere to biofilm attached to the pump. Results of the study indicate that bacteria may be accumulating and entrapping solid material on the exterior of pumps. This accumulation reduces heat transfer and water discharge from the pump and may lead to decreased efficiency or mechanical failure. Observations could not be made on the well screen, gravel pack, or surrounding geologic formation; therefore, mitigating measures in the borehole may not solve well-productivity issues. Remedies for the pump fouling problems were derived from the review and interpretation of data collected during this study and from information documented in other sources about groundwater well fouling. Potential remedies to problems associated with pump fouling at the CDF may include the following: (1) reducing attractiveness of the extraction wells for microbiological growth by modifying the chemical or physical environment of the well, (2) modifying the pump exterior to decrease microbiological adherence, (3) changing the pumping regime to control the chemistry of water entering the well from the surrounding aquifer material, (4) modifying the pumps to be less physically and thermally attractive, and (5) removing hydrocarbons from groundwater and the aquifer material surrounding the wells or adding surfactants to make them more mobile. Pilot scale testing may be necessary to identify the most effective treatment or combination of treatments.

Indiana

Methods for collection and analysis of aquatic biological and microbiological samples

The series of chapters on techniques describes methods used by the U.S. Geological Survey for planning and conducting water-resources investigations. The material is arranged under major subject headings called books and is further subdivided into sections and chapters. Book 5 is on laboratory analysis. Section A is on water. The unit of publication, the chapter, is limited to a narrow field of subject matter. "Methods for Collection and Analysis of Aquatic Biological and Microbiological Samples" is the fourth chapter to be published under Section A of Book 5. The chapter number includes the letter of the section. This chapter was prepared by several aquatic biologists and microbiologists of the U.S. Geological Survey to provide accurate and precise methods for the collection and analysis of aquatic biological and microbiological samples. Use of brand, firm, and trade names in this chapter is for identification purposes only and does not constitute endorsement by the U.S. Geological Survey. This chapter supersedes "Methods for Collection and Analysis of Aquatic Biological and Microbiological Samples" edited by P.E. Greeson, T.A. Ehlke, G.A. Irwin, B.W. Lium, and K.V. Slack (U.S. Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter A4, 1977) and also supersedes "A Supplement to-Methods for Collection and Analysis of Aquatic Biological and Microbiological Samples" by P.E. Greeson (U.S. Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter A4), Open-File Report 79-1279, 1979.

Techniques of Water-Resources Investigations

Natural attenuation of chlorinated-hydrocarbon contamination at Fort Wainwright, Alaska: A hydrogeochemical and microbiological investigation workplan

Natural attenuation processes include biological degradation, by which microorganisms break down contaminants into simpler product compounds; adsorption of contaminants to soil particles, which decreases the mass of contaminants dissolved in ground water; and dispersion, which decreases dissolved contaminant concentrations through dilution. The primary objectives of this study are to (1) assess the degree to which such natural processes are attenuating chlorinated-hydrocarbon contamination in ground water, and (2) evaluate the effects of ground-water/surface-water interactions on natural-attenuation processes in the area of the former East and West Quartermasters Fueling Systems for Fort Wainwright, Alaska. The study will include investigations of the hydrologic, geochemical, and microbiological processes occurring at this site that influence the transport and fate of chlorinated hydrocarbons in ground water. To accomplish these objectives, a data-collection program has been initiated that includes measurements of water-table elevations and the stage of the Chena River; measurements of vertical temperature profiles within the subsurface; characterization of moisture distribution and movement in the unsaturated zone; collection of ground-water samples for determination of both organic and inorganic chemical constituents; and collection of ground-water samples for enumeration of microorganisms and determination of their potential to mineralize contaminants. We will use results from the data-collection program described above to refine our conceptual model of hydrology and contaminant attenuation at this site. Measurements of water-table elevations and river stage will help us to understand the magnitude and direction of ground-water flow and how changes in the stage of the Chena River affect ground-water flow. Because ambient ground water and surface water typically have different temperature characteristics, temperature monitoring will likely provide further insight into ground-water/surface-water interactions in the subsurface. Characterization of the unsaturated zone will improve our understanding of interactions among ground water, the unsaturated zone, and the atmosphere. The interactions likely of importance to this study include the migration of water, dissolved contaminants, nutrients, and gases (oxygen, carbon dioxide, and methane) between the saturated and unsaturated zones. We will use the results of ground-water chemical analyses to determine the spatial and temporal distribution of (1) chlorinated-hydrocarbon contaminants and their degradation products, (2) oxidation-reduction indicators, (3) nutrients, and (4) major ground-water ions. These water-quality data will provide insight into ground-water flow directions, interactions between ground water and surface water, attenuation of contaminant concentrations caused by dispersion, and intrinsic microbiological processes. Microbiological analyses will indicate whether microorganisms at the site are capable of degrading the contaminants of interest, and will allow us to estimate their potential to attenuate existing contamination. Physical and chemical data interpreted as part of the analysis of ground water and surface water mixing will improve our understanding of the relationship between water quality and contaminant source mixing.

Alaska

Microbiological Water Quality in Relation to Water-Contact Recreation, Cuyahoga River, Cuyahoga Valley National Park, Ohio, 2000 and 2002

The microbiological water quality of a 23-mile segment of the Cuyahoga River within the Cuyahoga Valley National Park was examined in this study. This segment of the river receives discharges of contaminated water from stormwater, combined-sewer overflows, and incompletely disinfected wastewater. Frequent exceedances of Ohio microbiological water-quality standards result in a health risk to the public who use the river for water-contact recreation. Water samples were collected during the recreational season of May through October at four sites on the Cuyahoga River in 2000, at three sites on the river in 2002, and from the effluent of the Akron Water Pollution Control Station (WPCS) both years. The samples were collected over a similar range in streamflow in 2000 and 2002. Samples were analyzed for physical and chemical constituents, as well as the following microbiological indicators and pathogenic organisms: Escherichia coli (E. coli), Salmonella, F-specific and somatic coliphage, enterovirus, infectious enterovirus, hepatitis A virus, Clostridium perfringens (C. perfringens), Cryptosporidium, and Giardia. The relations of the microorganisms to each other and to selected water-quality measures were examined. All microorganisms analyzed for, except Cryptosporidium, were detected at least once at each sampling site. Concentrations of E. coli exceeded the Ohio primary-contact recreational standard (298 colonies per 100 milliliters) in approximately 87 percent of the river samples and generally were higher in the river samples than in the effluent samples. C. perfringens concentrations were positively and significantly correlated with E. coli concentrations in the river samples and generally were higher in the effluent samples than in the river samples. Several of the river samples that met the Ohio E. coli secondary-contact recreational standard (576 colonies per 100 milliliters) had detections of enterovirus, infectious enterovirus, hepatitis A virus, and Salmonella, indicating that there are still risks even when the E. coli standard is not exceeded. River samples in which the secondary-contact recreational standard for E. coli was exceeded showed a higher percentage of the co-occurrence of pathogenic organisms than samples that met the standard. This indicates that in this study area, E. coli is a useful indicator of human health risk. Detections of hepatitis A virus tended to be associated with higher median concentrations of somatic coliphage, F-specific coliphage, and infectious enterovirus. In addition, geometric mean C. perfringens concentrations tended to be higher in samples where hepatitis A virus was present than in samples where hepatitis A virus was absent. Hepatitis A virus was not detected in samples collected upstream from the Akron WPCS; all downstream detections had coincident detections in the Akron WPCS effluent, suggesting that Akron WPCS was a principal source of hepatitis A virus at the downstream sites. Geometric mean concentrations of E. coli were calculated on the basis of analytical results from at least five samples collected at each river site during May, July, and September of 2000. In each case, the Ohio geometric-mean primary-contact recreational standard of 126 col/100 mL was exceeded. E. coli concentrations were significantly correlated with streamflow and increased with streamflow at sites upstream and downstream from the Akron WPCS. This indicates that E. coli loads from sources upstream from the Akron WPCS have the potential to appreciably influence the frequency of attainment of recreational water-quality standards at downstream locations.

Water-Resources Investigations Report

Ground-water microbiology and geochemistry

Up-to-date coverage and a unique, multidisciplinary approach The ongoing effort to protect our valuable ground-water resources necessarily involves scientists and engineers from many disciplines. Ground-Water Microbiology and Geochemistry, Second Edition is designed to bridge the historical lack of communication among these disciplines by detailing-in language that cuts across specialties-the impact of microorganisms and microbial processes on ground-water systems. Carefully revised to reflect the many recent discoveries that have been made in the field, the Second Edition begins with an overview of microbiology, ideal for hydrologists and others who may lack formal training in the field. These initial chapters systematically cover the kinds of microorganisms found in subsurface environments, focusing on their growth, metabolism, genetics, and ecology. The second part of the book offers a hydrologic perspective on how microbial processes affect ground-water geochemistry in pristine systems. It also introduces the different classes of ground-water systems, and gives an overview of techniques for sampling subsurface environments. Readers gain an understanding of biogeochemical cycling in ground-water systems-in coverage unique to this book-and how ground-water chemistry can be used to study microbial processes in aquifer systems. The final section of the book deals with the biodegradation of human-introduced contaminants in ground-water systems, with an up-to-date review of the physiology, biochemistry, and redox conditions that favor biodegradation processes. Ground-Water Microbiology and Geochemistry, Second Edition is important reading for geoscientists, hydrologists, and environmental engineers, as well as for water planners and lawyers involved in environmental issues. It also serves as a compelling text for upper-level undergraduate and graduate courses in ground-water chemistry.

Book

Microbiological reduction of Sb(V) in anoxic freshwater sediments

Microbiological reduction of millimolar concentrations of Sb(V) to Sb(III) was observed in anoxic sediments from two freshwater settings: (1) a Sb- and As-contaminated mine site (Stibnite Mine) in central Idaho and 2) an uncontaminated suburban lake (Searsville Lake) in the San Francisco Bay Area. Rates of Sb(V) reduction in anoxic sediment microcosms and enrichment cultures were enhanced by amendment with lactate or acetate as electron donors but not by H2, and no reduction occurred in sterilized controls. Addition of 2- 14 C-acetate to Stibnite Mine microcosms resulted in the production of 14 CO 2 coupled to Sb(V) reduction, suggesting that this process proceeds by a dissimilatory respiratory pathway in those sediments. Antimony(V) reduction in Searsville Lake sediments was not coupled to acetate mineralization and may be associated with Sb-resistance. The microcosms and enrichment cultures also reduced sulfate, and the precipitation of insoluble Sb(III)-sulfide complexes was a major sink for reduced Sb. The reduction of Sb(V) by Stibnite Mine sediments was inhibited by As(V), suggesting that As(V) is a preferred electron acceptor for the indigenous community. These findings indicate a novel pathway for anaerobic microbiological respiration and suggest that communities capable of reducing high concentrations of Sb(V) commonly occur naturally in the environment.

Environmental Science & Technology

Advances in the hydrogeochemistry and microbiology of acid mine waters

The last decade has witnessed a plethora of research related to the hydrogeochemistry and microbiology of acid mine waters and associated tailings and waste-rock waters. Numerous books, reviews, technical papers, and proceedings have been published that examine the complex bio-geochemical process of sulfide mineral oxidation, develop and apply geochemical models to site characterization, and characterize the microbial ecology of these environments. This review summarizes many of these recent works, and provides references for those investigating this field. Comparisons of measured versus calculated Eh and measured versus calculated pH for water samples from several field sites demonstrate the reliability of some current geochemical models for aqueous speciation and mass balances. Geochemical models are not, however, used to predict accurately time-dependent processes but to improve our understanding of these systems and to constrain possible processes that contribute to actual or potential water quality issues. Microbiological studies are demonstrating that there is much we have yet to learn about the types of different microorganisms and their function and ecology in mine-waste environments. A broad diversity of green algae, bacteria, archaea, yeasts, and fungi are encountered in acid mine waters, and a better understanding of their ecology and function may potentially enhance remediation possibilities as well as our understanding of the evolution of life.

International Geology Review

Overview of issues in subsurface and landfill microbiology

To date, the majority of evidence indicates that most subsurface environments possess climax ecological communities that are well adapted to the environment in which they live. Like their counterparts on the surface, subsurface ecosystems are characterized by a high degree of microbiological diversity, they possess trophic structure, and they exhibit material cycling and energy transfer. Members of such ecosystems typically possess structural, physiological, or reproductive adaptations that allow them to disperse and survive in such habitats. Current studies argue that microbial activities influence the geochemical processes in both landfills and subsurface environments and that the altered geochemistry, in turn, influences the selection of microorganisms proliferating in the habitat. However, while microbial activity may often be limited by the availability of electron donors in uncontaminated aquifer systems, this is rarely the case with landfills. Sections in this chapter provide investigators with a greater understanding of the experimental approaches needed to study the microbiology of the terrestrial subsurface and an appreciation of interpretational limits imposed by the existing methodologies. The hope is that further study of subsurface microorganisms will provide insights into the process of microbial evolution and possibly into the origins of life itself.

Book chapter

Occurrence and distribution of microbiological indicators in groundwater and stream water

A total of 136 stream water and 143 groundwater samples collected in five important hydrologic systems of the United States were analyzed for microbiological indicators to test monitoring concepts in a nationally consistent program. Total coliforms were found in 99%, Escherichia coli in 97%, and Clostridium perfringens in 73% of stream water samples analyzed for each bacterium. Total coliforms were found in 20%, E. coli in less than 1%, and C. perfringens in none of the groundwater samples analyzed for each bacterium. Although coliphage analyses were performed on many of the samples, contamination in the laboratory and problems discerning discrete plaques precluded quantification. Land use was found to have the most significant effect on concentrations of bacterial indicators in stream water. Presence of septic systems on the property near the sampling site and well depth were found to be related to detection of coliforms in groundwater, although these relationships were not statistically significant. A greater diversity of sites, more detailed information about some factors, and a larger dataset may provide further insight to factors that affect microbiological indicators.

Water Environment Research

Initiation of long-term coupled microbiological, geochemical, and hydrological experimentation within the seafloor at North Pond, western flank of the Mid-Atlantic Ridge

Integrated Ocean Drilling Program (IODP) Expedition 336 successfully initiated subseafloor observatory science at a young mid-ocean-ridge flank setting. All of the drilled sites are located in the North Pond region of the Atlantic Ocean (22°45′N, 46°05′W) in 4414–4483 m water depth. This area is known from previous ocean drilling and site survey investigations as a site of particularly vigorous circulation of seawater in permeable 8 Ma basaltic basement underlying a <300 m thick sedimentary pile. Understanding how this seawater circulation affects microbial and geochemical processes in the uppermost basement was the primary science objective of Expedition 336. Basement was cored and wireline-logged in Holes U1382A and U1383C. Upper oceanic crust in Hole U1382A, which is only 50 m west of Deep Sea Drilling Project (DSDP) Hole 395A, recovered 32 m of core between 110 and 210 meters below seafloor (mbsf). Core recovery in basement was 32%, yielding a number of volcanic flow units with distinct geochemical and petrographic characteristics. A unit of sedimentary breccia containing clasts of basalt, gabbroic rocks, and mantle peridotite was found intercalated between two volcanic flow units and was interpreted as a rock slide deposit. From Hole U1383C we recovered 50.3 m of core between 69.5 and 331.5 mbsf (19%). The basalts are aphyric to highly plagioclase-olivine-phyric tholeiites that fall on a liquid line of descent controlled by olivine fractionation. They are fresh to moderately altered, with clay minerals (saponite, nontronite, and celadonite), Fe oxyhydroxide, carbonate, and zeolite as secondary phases replacing glass and olivine to variable extents. In addition to traditional downhole logs, we also used a new logging tool for detecting in situ microbial life in ocean floor boreholes—the Deep Exploration Biosphere Investigative tool (DEBI-t). Sediment thickness was ~90 m at Sites U1382 and U1384 and varied between 38 and 53 m at Site U1383. The sediments are predominantly nannofossil ooze with layers of coarse foraminiferal sand and occasional pebble-size clasts of basalt, serpentinite, gabbroic rocks, and bivalve debris. The bottommost meters of sections cored with the advanced piston corer feature brown clay. Extended core barrel coring at the sediment/ basement interface recovered <1 m of brecciated basalt with micritic limestone. Sediments were intensely sampled for geochemical pore water analyses and microbiological work. In addition, high-resolution measurements of dissolved oxygen concentration were performed on the whole-round sediment cores Major strides in ridge-flank studies have been made with subseafloor borehole observatories (CORKs) because they facilitate combined hydrological, geochemical, and microbiological studies and controlled experimentation in the subseafloor. During Expedition 336, two fully functional observatories were installed in two newly drilled holes (U1382A and U1383C) and an instrument and sampling string were placed in an existing hole (395A). Although the CORK wellhead in Hole 395A broke off and Hole U1383B was abandoned after a bit failure, these holes and installations are intended for future observatory science targets. The CORK observatory in Hole U1382A has a packer seal in the bottom of the casing and monitors/samples a single zone in uppermost oceanic crust extending from 90 to 210 mbsf. Hole U1383C was equipped with a three-level CORK observatory that spans a zone of thin basalt flows with intercalated limestone (~70–146 mbsf), a zone of glassy, thin basaltic flows and hyaloclastites (146–200 mbsf), and a lowermost zone (~200–331.5 mbsf) of more massive pillow flows with occasional hyaloclastites in the upper part.

Integrated Ocean Drilling Program: Preliminary Rep

The U.S. Geological Survey Ohio Water Microbiology Laboratory

The U.S. Geological Survey Ohio Water Microbiology Laboratory is a part of the Ohio-Kentucky-Indiana Water Science Center. The mission of the laboratory is to provide microbiological data of public health significance from surface waters, groundwaters, and sediments for a variety of study objectives. The laboratory conducts internal projects, works with external cooperators, and assists U.S. Geological Survey offices and National programs. The laboratory offers guidance, study design, and data interpretation expertise to collaborators, all following rigorous quality control and quality assurance procedures.

Fact Sheet

A supplement to "Methods for collection and analysis of aquatic biological and microbiological samples" (U.S. Geological Survey Techniques of Water-Resources Investigations, Book 5, Chapter A4)

The manual 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 are 5 new methods, a new section of selected taxonomic references for Ostracoda, and 6 revised methods.

Open-File Report

Methods for collection and analysis of aquatic biological and microbiological samples

Chapter A4, methods for collection and analyses of aquatic biological and microbiological samples, contains methods used by the U.S. Geological Survey to collect, preserve, and analyze waters to determine their biological and microbiological properties. Part 1 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 bioassay. Each method is summarized, and the applications, interferences, apparatus, reagents, analyses, calculations, reporting of results, precisions, and references are given. Part 2 consists of a glossary. Part 3 is a list of taxonomic references. (USGS)

Open-File Report

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