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USGS2004

Microbial precipitation of dolomite in methanogenic groundwater

We report low-temperature microbial precipitation of dolomite in dilute natural waters from both field and laboratory experiments. In a freshwater aquifer, microorganisms colonize basalt and nucleate nonstoichiometric dolomite on cell walls. In the laboratory, ordered dolomite formed at near-equilibrium conditions from groundwater with molar Mg:Ca ratios of <1; dolomite was absent in sterile experiments. Geochemical and microbiological data suggest that methanogens are the dominant metabolic guild in this system and are integral to dolomite precipitation. We hypothesize that the attached microbial consortium reacts with the basalt surface, releasing Mg and Ca into solution, which drives dolomite precipitation via nucleation on the cell wall. These findings provide insight into the long-standing dolomite problem and suggest a fundamental role for microbial processes in the formation of dolomite across a wide range of environmental conditions.

Geology↗

USGS2004

Evaluation of Water Quality for Two St. Johns River Tributaries Receiving Septic Tank Effluent, Duval County, Florida

Tributary streamflow to the St. Johns River in Duval County is thought to be affected by septic tank leachate from residential areas adjacent to these tributaries. Water managers and the city of Jacksonville have committed to infrastructure improvements as part of a management plan to address the impairment of tributary water quality. In order to provide data to evaluate the effects of future remedial activities in selected tributaries, major ion and nutrient concentrations, fecal coliform concentrations, detection of wastewater compounds, and tracking of bacterial sources were used to document septic tank influences on the water quality of selected tributaries. The tributaries Fishing Creek and South Big Fishweir Creek were selected because they drain subdivisions identified as high priority locations for septic tank phase-out projects: the Pernecia and Murray Hill B subdivisions, respectively. Population, housing (number of residences), and septic tank densities for the Murray Hill B subdivision are greater than those for the Pernecia subdivision. Water-quality samples collected in the study basins indicate influences from ground water and septic tanks. Estimated concentrations of total nitrogen ranged from 0.33 to 2.86 milligrams per liter (mg/L), and ranged from less than laboratory reporting limit (0.02 mg/L) to 0.64 mg/L for total phosphorus. Major ion concentrations met the State of Florida Class III surface-water standards; total nitrogen and total phosphorus concentrations exceeded the U.S. Environmental Protection Agency Ecoregion XII nutrient criteria for rivers and streams 49 and 96 percent of the time, respectively. Organic wastewater compounds detected at study sites were categorized as detergents, antioxidants and flame retardants, manufactured polycarbonate resins, industrial solvents, and mosquito repellent. The most commonly detected compound was para-nonylphenol, a breakdown product of detergent. Results of wastewater sampling give evidence that stream water in the study basins is affected by septic tank effluent. Fecal coliform bacteria concentrations were measured on a monthly basis; of 115 samples, 63 percent exceeded the State of Florida fecal coliform bacteria standard for Class III surface waters of 800 colonies per 100 milliliters of water on any 1 day. Fecal coliform bacteria concentrations ranged from less than 20 colonies per 100 milliliters of sample to greater than or equal to 160,000 colonies per 100 milliliters of sample. Antibiotic resistance patterns of fecal coliform bacteria were used to identify the sources of fecal coliform bacteria. Significant sources of fecal coliform bacteria included wild animals, dogs, and humans. A majority of the fecal coliform bacteria were classified to be from human sources. Because the primary source of fecal coliform bacteria is from human sources, and most likely septic tank effluent, management of human sources may substantially improve microbiological water quality in both the Fishing Creek and South Branch Big Fishweir Creek basins.

Water-Resources Investigations Report↗

USGS2017

Environmental Escherichia coli : Ecology and public health implications - A review

Escherichia coli is classified as a rod-shaped, Gram-negative bacterium in the family Enterobacteriaceae . The bacterium mainly inhabits the lower intestinal tract of warm-blooded animals, including humans, and is often discharged into the environment through feces or wastewater effluent. The presence of E. coli in environmental waters has long been considered as an indicator of recent fecal pollution. However, numerous recent studies have reported that some specific strains of E. coli can survive for long periods of time, and potentially reproduce, in extra-intestinal environments. This indicates that E. coli can be integrated into indigenous microbial communities in the environment. This naturalization phenomenon calls into question the reliability of E. coli as a fecal indicator bacterium (FIB). Recently, many studies reported that E. coli populations in the environment are affected by ambient environmental conditions affecting their long-term survival. Large-scale studies of population genetics provide the diversity and complexity of E. coli strains in various environments, affected by multiple environmental factors. This review examines the current knowledge on the ecology of E. coli strains in various environments in regards to its role as a FIB and as a naturalized member of indigenous microbial communities. Special emphasis is given on the growth of pathogenic E. coli in the environment, and the population genetics of environmental members of the genus Escherichia . The impact of environmental E. coli on water quality and public health is also discussed.

Journal of Applied Microbiology↗

USGS2001

Green fluorescent protein is lighting up fungal biology

Prasher ( 42 ) cloned a cDNA for the green fluorescent protein (GFP) gene from the jellyfish Aequorea victoria in 1992. Shortly thereafter, to the amazement of many investigators, this gene or derivatives thereof were successfully expressed and conferred fluorescence to bacteria and Caenorhabditis elegans cells in culture ( 10 , 31 ), followed by yeast ( 24 , 39 ), mammals ( 40 ), Drosophila ( 66 ), Dictyostelium ( 23 , 30 ), plants ( 28 , 49 ), and filamentous fungi ( 54 ). The tremendous success of GFP as a reporter can be attributed to unique qualities of this 238-amino-acid, 27-kDa protein which absorbs light at maxima of 395 and 475 nm and emits light at a maximum of 508 nm. The fluorescence of GFP requires only UV or blue light and oxygen, and therefore, unlike the case with other reporters (β-glucuronidase, β-galacturonidase, chloramphenicol acetyltransferase, and firefly luciferase) that rely on cofactors or substrates for activity, in vivo observation of gfp expression is possible with individual cells, with cell populations, or in whole organisms interacting with symbionts or environments in real time. Complications caused by destructive sampling, cell permeablization for substrates, or leakage of products do not occur. Furthermore, the GFP protein is extremely stable in vivo and has been fused to the C or N terminus of many cellular and extracellular proteins without a loss of activity, thereby permitting the tagging of proteins for gene regulation analysis, protein localization, or specific organelle labeling. The mature protein resists many proteases and is stable up to 65°C and at pH 5 to 11, in 1% sodium dodecyl sulfate or 6 M guanidinium chloride (reviewed in references 17 and 67 ), and in tissue fixed with formaldehyde, methanol, or glutaraldehyde. However, GFP loses fluorescence in methanol-acetic acid (3:1) and can be masked by autofluorescent aldehyde groups in tissue fixed with glutaraldehyde. Fluorescence is optimal at pH 7.2 to 8.0 ( 67 ). Limitations on GFP as a reporter for some applications are its low turnover rate, 2-h lag time for autoactivation of its chromophore, improper folding at high temperatures (37°C), which results in nonfluorescent and insoluble forms of the protein, and requirement for oxygen, which is not present in equal concentrations in all subcellular locations or cell types (reviewed in references 17 and 67 ). These characteristics of GFP, however, have not posed a problem for many applications, and mutant forms of GFP that have an ability to fold properly at high temperatures, increased solubility and fluorescence, reduced photobleaching ( 16 , 17 , 51 ), and reduced half-lives ( 1 ) have been developed. Coupled with fluorescence-activated cell sorting, confocal microscopy or quantitative image analysis techniques, GFP technology can be used to isolate transformed cells or specific cell types from populations of cells ( 14 ), to quantify gene expression of individual cells within whole organisms ( 8 ), or to assess the dispersal and biomass of organisms in complex environments, such as in animal or plant hosts ( 38 , 59 ), in biofilms ( 55 ), in fermentors ( 41 ), on leaf surfaces ( 53 , 61 ), or in soils ( 2 ). The vast majority of studies utilizing GFP expression in fungi have been with yeast (reviewed in reference 13 ). Ustilago maydis was the first filamentous fungus for which successful expression of gfp was reported ( 54 ), followed closely by Aspergillus nidulans ( 22 , 57 ) and Aureobasidium pullulans ( 61 ). Presently, gfp expression has been reported for 16 species comprising 12 genera of filamentous fungi, including Colletotrichum ( 21 , 44 ), Mycosphaerella ( 52 ), Magnaporthe ( 32 , 35 ), Cochliobolus ( 38 ), Trichoderma ( 2 , 70 ), Podospora ( 5 ), Sclerotinia ( 63 ), Schizophyllum ( 37 ), Aspergillus ( 20 , 47 , 50 ) and Phytophthora ( 7 , 62 ). In this review we draw on published reports, with the goal of providing an overview of GFP technology as it applies to the biology of filamentous fungi. These reports are not exhaustive of potential applications of GFP technology, as examples of genomic approaches to utilizing GFP in bacterial and yeast systems attest ( 4 , 46 , 60 , 65 ). Expression of gfp in filamentous fungi requires a gfp variant that is efficiently translated in fungi, a transformation system, and a fungal promoter that satisfies the requirements of a given experimental objective. Transformation of fungi has recently been reviewed by Gold et al. ( 26 ). Robinson and Sharon ( 44 ) suggest that GFP can actually be used to optimize transformation protocols. In addition to reporting the construction of a new fungal transformation vector that expresses SGFP under the control of the ToxA gene promoter from Pyrenophora tritici-repentis ( 12 ) and demonstrating its use in plant pathogens belonging to eight different genera of filamentous fungi ( Fusarium, Botrytis, Pyrenophora, Alternaria, Cochliobolus, Sclerotinia, Colletotrichum , and Verticillium ), in this review we also enumerate and describe a comprehensive list of vectors for expressing GFP in fungi.

Applied and Environmental Microbiology↗

USGS2021

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

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

Data Series↗

USGS2005

Bacteriological water quality in the Lake Pontchartrain basin, Louisiana, following Hurricanes Katrina and Rita, September 2005

The U.S. Geological Survey (USGS), in collaboration with the Louisiana Department of Environmental Quality, monitored bacteriological quality of water at 22 sites in and around Lake Pontchartrain, La., for three consecutive weeks beginning September 13, 2005, following hurricanes Katrina and Rita and the associated flooding. Samples were collected and analyzed by USGS personnel from the USGS Louisiana Water Science Center and the USGS Ohio Water Microbiology Laboratory. Fecal-indicator bacteria (Escherichia coli, enterococci, and fecal coliform) concentrations ranged from the detection limit to 36,000 colony-forming units per 100 milliliters. Data are presented in tabular form and as plots of data in the context of available historical data and water-quality standards and criteria for each site sampled. Quality-control data were reviewed to ensure that methods performed as expected in a mobile laboratory setting.

Louisiana↗

USGS1984

Water quality of a stream-aquifer system, southern Franklin County, Ohio

The chemical quality of the water resources in the Scioto River valley south of Columbus, Ohio, was evaluated on the basis of data collected from 74 wells and 4 surface-water sites. A very hard calcium bicarbonate water that is high in dissolved solids is characteristic of the aquifer. Mean pH of the ground water is 7.3, and bicaerbonate concentrations range from 238 to 530 milligrams per liter. Concentrations of dissolved iron in water samples (0.01 to 3.9 milligrams per liter) frequently exceed the drinking water standard of 0.3 milligrams per liter established by the Ohio Environmental Protection Agency. The chemical quality of local streams closely resembles the ground-water quality, except for higher concentrations of sodium, chloride, and sulfate in the surface water. Microbiological testing of ground water for fecal bacteria indicated concentrations ranging from 1 to 2,400 colonies per 100 milliliters. The higher concentrations were observed in one well on the Scioto River flood plain after a flood event. Mass-balance calculations of the chemical data indicate that pH is buffered primarily by the carbonate system. Data for pH, calculated Eh, and concentrations of ferrous iron and reduced sulfur show that areas of different chemical environments exist within the aquifer. A reaction model of an induced-infiltration radial collector system indicates the amount of mass transfer (chemical reactions that occur as ground water and surface water mix) is small, and that the mixing ratio between ground and surface waters is four to one.

Water-Resources Investigations Report↗

USGS2008

Water-quality effects and characterization of indicators of onsite wastewater disposal systems in the east-central Black Hills area, South Dakota, 2006-08

Onsite wastewater disposal systems (OWDS) are used extensively in the Black Hills of South Dakota where many of the watersheds and aquifers are characterized by fractured or solution-enhanced bedrock with thin soil cover. A study was conducted during 2006-08 to characterize water-quality effects and indicators of OWDS. Water samples were collected and analyzed for potential indicators of OWDS, including chloride, bromide, boron, nitrite plus nitrate (NO2+NO3), ammonia, major ions, nutrients, selected trace elements, isotopes of nitrate, microbiological indicators, and organic wastewater compounds (OWCs). The microbiological indicators were fecal coliforms, Escherichia coli (E. coli), enterococci, Clostridium perfringens (C. perfringens), and coliphages. Sixty ground-water sampling sites were located either downgradient from areas of dense OWDS or in background areas and included 25 monitoring wells, 34 private wells, and 1 spring. Nine surface-water sampling sites were located on selected streams and tributaries either downstream or upstream from residential development within the Precambrian setting. Sampling results were grouped by their hydrogeologic setting: alluvial, Spearfish, Minnekahta, and Precambrian. Mean downgradient dissolved NO2+NO3 concentrations in ground water for the alluvial, Spearfish, Minnekahta, and Precambrian settings were 0.734, 7.90, 8.62, and 2.25 milligrams per liter (mg/L), respectively. Mean downgradient dissolved chloride concentrations in ground water for these settings were 324, 89.6, 498, and 33.2 mg/L, respectively. Mean downgradient dissolved boron concentrations in ground water for these settings were 736, 53, 64, and 43 micrograms per liter (ug/L), respectively. Mean dissolved surface-water concentrations for NO2+NO3, chloride, and boron for downstream sites were 0.222 mg/L, 32.1 mg/L, and 28 ug/L, respectively. Mean values of delta-15N and delta-18O (isotope ratios of 14N to 15N and 18O to 16O relative to standard ratios) for nitrate in ground-water samples were 10.4 and -2.0 per mil (0/100), respectively, indicating a relatively small contribution from synthetic fertilizer and probably a substantial contribution from OWDS. The surface-water sample with the highest dissolved NO2+NO3 concentration of 1.6 mg/L had a delta-15N value of 12.36 0/100, which indicates warm-blooded animals (including humans) as the nitrate source. Fecal coliforms were detected in downgradient ground water most frequently in the Spearfish (19 percent) and Minnekahta (9.7 percent) settings. E. coli was detected most frequently in the Minnekahta (29 percent) and Spearfish (13 percent) settings. Enterococci were detected more frequently than other microbiological indicators in all four settings. Fecal coliforms and E. coli were detected in 73 percent and 95 percent of all surface-water samples, respectively. Enterococci, coliphages (somatic), and C. perfringens were detected in 50, 70, and 50 percent of surface-water samples, respectively. Of the 62 OWC analytes, 12 were detected only in environmental samples, 10 were detected in at least one environmental and one blank sample (not necessarily companion pairs), 2 were detected only in blank samples, and 38 were not detected in any blank, environmental, or replicate sample from either ground or surface water. Eleven different organic compounds were detected in ground-water samples at eight different sites. The most frequently occurring compound was DEET, which was found in 32 percent of the environmental samples, followed by tetrachloroethene, which was detected in 20 percent of the samples. For surface-water samples, 16 organic compounds were detected in 9 of the 10 total samples. The compound with the highest occurrence in surface-water samples was camphor, which was detected in 50 percent of samples. The alluvial setting was characterized by relatively low dissolved NO2+NO3 concentrations, detection of ammonia nitrogen, and relatively high concentr

South Dakota↗

USGS2000

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↗

USGS2017

Potential for water borne and invertebrate transmission of West Nile virus in the Great Salt Lake, Utah

In November and December of 2013, a large mortality event involving 15,000 - 20,000 eared grebes ( Podiceps nigricollis ) occurred at the Great Salt Lake (GSL), UT. The onset of the outbreak in grebes was followed by a mortality event in > 86 bald eagles ( Haliaeetus leucocephalus ). During the die-off, West Nile virus (WNV) was detected by RT-PCR or viral culture in carcasses of grebes and eagles submitted to the National Wildlife Health Center. However, no mosquito activity, the primary vector of WNV, was detected by the State of Utah's WNV monitoring program. Transmission of WNV has rarely been reported during the winter in North America in the absence of known mosquito activity; however, the size of this die-off, the habitat in which it occurred, and the species involved are unique. We experimentally investigated whether WNV could survive in water with a high saline content, as found at the GSL, and whether brine shrimp, the primary food of migrating eared grebes on the GSL, could have played a role in transmission of WNV to feeding birds. We found that WNV can survive up to 72 h at 4°C in water containing 30 — 150 ppt NaCl and brine shrimp, incubated with WNV in 30 ppt NaCl, may adsorb WNV to their cuticle and, through feeding, may infect epithelial cells of their gut. Both mechanisms may have potentiated the WNV die-off in migrating eared grebes on the GSL.

Utah↗

USGS2025

STREAMS guidelines: Standards for technical reporting in environmental and host-associated microbiome studies

The interdisciplinary nature of microbiome research, coupled with the generation of complex multi-omics data, makes knowledge sharing challenging. The Strengthening the Organization and Reporting of Microbiome Studies (STORMS) guidelines provide a checklist for the reporting of study information, experimental design and analytical methods within a scientific manuscript on human microbiome research. Here, in this Consensus Statement, we present the standards for technical reporting in environmental and host-associated microbiome studies (STREAMS) guidelines. The guidelines expand on STORMS and include 67 items to support the reporting and review of environmental (for example, terrestrial, aquatic, atmospheric and engineered), synthetic and non-human host-associated microbiome studies in a standardized and machine-actionable manner. Based on input from 248 researchers spanning 28 countries, we provide detailed guidance, including comparisons with STORMS, and case studies that demonstrate the usage of the STREAMS guidelines. STREAMS, like STORMS, will be a living community resource updated by the Consortium with consensus-building input of the broader community.

Nature Microbiology↗

USGS2013

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

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

California↗

USGS2010

Coupled arsenotrophy in a hot spring photosynthetic biofilm at Mono Lake, California

Red-pigmented biofilms grow on rock and cobble surfaces present in anoxic hot springs located on Paoha Island in Mono Lake. The bacterial community was dominated (∼ 85% of 16S rRNA gene clones) by sequences from the photosynthetic Ectothiorhodospira genus. Scraped biofilm materials incubated under anoxic conditions rapidly oxidized As(III) to As(V) in the light via anoxygenic photosynthesis but could also readily reduce As(V) to As(III) in the dark at comparable rates. Back-labeling experiments with 73 As(V) demonstrated that reduction to 73 As(III) also occurred in the light, thereby illustrating the cooccurrence of these two anaerobic processes as an example of closely coupled arsenotrophy. Oxic biofilms also oxidized As(III) to As(V). Biofilms incubated with [ 14 C]acetate oxidized the radiolabel to 14 CO 2 in the light but not the dark, indicating a capacity for photoheterotrophy but not chemoheterotrophy. Anoxic, dark-incubated samples demonstrated As(V) reduction linked to additions of hydrogen or sulfide but not acetate. Chemoautotrophy linked to As(V) as measured by dark fixation of [ 14 C]bicarbonate into cell material was stimulated by either H 2 or HS − . Functional genes for the arsenate respiratory reductase ( arrA ) and arsenic resistance ( arsB ) were detected in sequenced amplicons of extracted DNA, with about half of the arrA sequences closely related (∼98% translated amino acid identity) to those from the family Ectothiorhodospiraceae . Surprisingly, no authentic PCR products for arsenite oxidase ( aoxB ) were obtained, despite observing aerobic arsenite oxidation activity. Collectively, these results demonstrate close linkages of these arsenic redox processes occurring within these biofilms. Oxyanions of the group 15 element arsenic, arsenate [As(V)] and arsenite [As(III)], have been known for millennia to be potent poisons. Despite its well-established toxicity to life, the phenomenon of arsenic resistance was discovered whereby some microorganisms maintain an otherwise “normal” existence in the presence of high concentrations of As(V) or As(III) (17, 29, 31). More recently it has become recognized that certain representatives from the bacterial and archaeal domains can actually exploit the electrochemical potential of the As(V)/As(III) redox couple (+130 mV) to gain energy for growth. This can be achieved either by employing As(III) as an autotrophic electron donor or by using As(V) as a respiratory electron acceptor (18, 21, 34). The latter phenomenon, although most commonly associated with chemoheterotrophy, can also employ inorganic substances like sulfide or H 2 . Indeed, As(V)-respiring anaerobes displaying a capacity for chemoautotrophy with these electron donors have been isolated and described (5, 7, 16). We recently reported that photoautotrophy is supported by As(III) in anoxic biofilms located in hot springs on Paoha Island in Mono Lake, CA (15). This process represented a novel means of As(III) oxidation achieved via anoxygenic photosynthesis occurring in certain photosynthetic bacteria (i.e., Ectothiorhodospira ) and possibly within some cyanobacteria as well (e.g., “ Oscillatoria ”). Whether or not a microbial habitat is overtly oxic or anoxic, or temporally shifts between these two states over a diel cycle, critical energy linkages between aerobes and anaerobes have long been known for the biogeochemical cycles of key elements, such as sulfur, iron, and nitrogen. Most prominently studied is the case of nitrogen, whereby an ecological coupling exists between the processes of nitrification and denitrification (9, 10, 28). The former process provides energy to aerobic nitrifiers, while the latter process consumes the nitrate produced by this reaction, thereby meeting the energy needs of the denitrifiers. For arsenic, the detection of both As(III) oxidation and As(V) reduction in oxic and anoxic incubations of freshly collected periphyton suggested that an analogous coupled process may also occur for this element (12). Similarly, several uncontaminated soils in Japan displayed a capacity for either As(V) reduction or As(III) oxidation upon arsenic oxyanion amendment and whether they were incubated under oxic or anoxic conditions (39). A defined coculture consisting of an aerobic As(III) oxidizer (strain OL1) and an anaerobic As(V) respirer (strain Y5) was shown to function in this fashion under manipulated laboratory conditions of oxygen tension (26). We pursued the phenomenon of coupled arsenic metabolism further by using materials collected from the hot spring biofilms in Mono Lake, but we focused on examination of the cycling of arsenic under anoxic conditions. In this paper we report results obtained by manipulated incubations of red-pigmented biofilms found in the hot springs of Paoha Island. Preliminary community characterizations of these biofilms show that they are dominated by B acteria from the genus Ectothiorhodospira but also harbor an assemblage of Archaea related to the Halobacteriacaea . Incubation results have demonstrated the presence of the following arsenic metabolic activities: respiratory As(V) reduction, photosynthetic anaerobic As(III) oxidation, and aerobic As(III) oxidation, along with the ecophysiological conditions under which they occur. Surprisingly, we were unable to obtain authentic PCR products for arsenite oxidase genes ( aoxB ), despite observing aerobic As(III) oxidation activity. These biofilms serve as a model system for how anaerobic cycling of arsenic can be sustained with oxidation of As(III) by anoxygenic photosynthesis coupled to regeneration of this electron donor via dissimilatory As(V) reduction. The significance that such a light-driven anaerobic ecosystem may have played in the Archean Earth is discussed.

California↗

USGS2016

Developing integrated methods to address complex resource and environmental issues

Introduction This circular provides an overview of selected activities that were conducted within the U.S. Geological Survey (USGS) Integrated Methods Development Project, an interdisciplinary project designed to develop new tools and conduct innovative research requiring integration of geologic, geophysical, geochemical, and remote-sensing expertise. The project was supported by the USGS Mineral Resources Program, and its products and acquired capabilities have broad applications to missions throughout the USGS and beyond. In addressing challenges associated with understanding the location, quantity, and quality of mineral resources, and in investigating the potential environmental consequences of resource development, a number of field and laboratory capabilities and interpretative methodologies evolved from the project that have applications to traditional resource studies as well as to studies related to ecosystem health, human health, disaster and hazard assessment, and planetary science. New or improved tools and research findings developed within the project have been applied to other projects and activities. Specifically, geophysical equipment and techniques have been applied to a variety of traditional and nontraditional mineral- and energy-resource studies, military applications, environmental investigations, and applied research activities that involve climate change, mapping techniques, and monitoring capabilities. Diverse applied geochemistry activities provide a process-level understanding of the mobility, chemical speciation, and bioavailability of elements, particularly metals and metalloids, in a variety of environmental settings. Imaging spectroscopy capabilities maintained and developed within the project have been applied to traditional resource studies as well as to studies related to ecosystem health, human health, disaster assessment, and planetary science. Brief descriptions of capabilities and laboratory facilities and summaries of some applications of project products and research findings are included in this circular. The work helped support the USGS mission to “provide reliable scientific information to describe and understand the Earth; minimize loss of life and property from natural disasters; manage water, biological, energy, and mineral resources; and enhance and protect our quality of life.” Activities within the project include the following: Spanned scales from microscopic to planetary; Demonstrated broad applications across disciplines; Included life-cycle studies of mineral resources; Incorporated specialized areas of expertise in applied geochemistry including mineralogy, hydrogeology, analytical chemistry, aqueous geochemistry, biogeochemistry, microbiology, aquatic toxicology, and public health; and Incorporated specialized areas of expertise in geophysics including magnetics, gravity, radiometrics, electromagnetics, seismic, ground-penetrating radar, borehole radar, and imaging spectroscopy. This circular consists of eight sections that contain summaries of various activities under the project. The eight sections are listed below: Laboratory Facilities and Capabilities, which includes brief descriptions of the various types of laboratories and capabilities used for the project; Method and Software Development, which includes summaries of remote-sensing, geophysical, and mineralogical methods developed or enhanced by the project; Instrument Development, which includes descriptions of geophysical instruments developed under the project; Minerals, Energy, and Climate, which includes summaries of research that applies to mineral or energy resources, environmental processes and monitoring, and carbon sequestration by earth materials; Element Cycling, Toxicity, and Health, which includes summaries of several process-oriented geochemical and biogeochemical studies and health-related research activities; Hydrogeology and Water Quality, which includes descriptions of innovative geophysical, remote-sensing, and geochemical research pertaining to hydrogeology and water-quality applications; Hazards and Disaster Assessment, which includes summaries of research and method development that were applied to natural hazards, human-caused hazards, and disaster assessments; and Databases and Framework Studies, which includes descriptions of fundamental applications of geophysical studies and of the importance of archived data.

Circular↗

USGS2016

Functional metagenomic selection of RubisCOs from uncultivated bacteria

Ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) is a critical yet severely inefficient enzyme that catalyses the fixation of virtually all of the carbon found on Earth. Here, we report a functional metagenomic selection that recovers physiologically active RubisCO molecules directly from uncultivated and largely unknown members of natural microbial communities. Selection is based on CO 2 -dependent growth in a host strain capable of expressing environmental deoxyribonucleic acid (DNA), precluding the need for pure cultures or screening of recombinant clones for enzymatic activity. Seventeen functional RubisCO-encoded sequences were selected using DNA extracted from soil and river autotrophic enrichments, a photosynthetic biofilm and a subsurface groundwater aquifer. Notably, three related form II RubisCOs were recovered which share high sequence similarity with metagenomic scaffolds from uncultivated members of the G allionellaceae family. One of the G allionellaceae RubisCOs was purified and shown to possess CO 2 /O 2 specificity typical of form II enzymes. X-ray crystallography determined that this enzyme is a hexamer, only the second form II multimer ever solved and the first RubisCO structure obtained from an uncultivated bacterium. Functional metagenomic selection leverages natural biological diversity and billions of years of evolution inherent in environmental communities, providing a new window into the discovery of CO 2 -fixing enzymes not previously characterized.

Environmental Microbiology↗

USGS2023

A cross inoculation experiment reveals Ophidiomyces ophiodiicola and Nannizziopsis guarroi can each infect both snakes and lizards

Host range and specificity are key concepts in the study of infectious diseases. However, both concepts remain largely undefined for many influential pathogens, including many fungi within the Onygenales order. This order encompasses reptile-infecting genera ( Nannizziopsis , Ophidiomyces , and Paranannizziopsis ) formerly classified as the Chrysosporium anamorph of Nannizziopsis vriesii (CANV). The reported hosts of many of these fungi represent a narrow range of phylogenetically related animals, suggesting that many of these disease-causing fungi are host specific, but the true number of species affected by these pathogens is unknown. For example, to date, Nannizziopsis guarroi (the causative agent of yellow fungus disease) and Ophidiomyces ophiodiicola (the causative agent of snake fungal disease) have been documented only in lizards and snakes, respectively. In a 52-day reciprocal-infection experiment, we tested the ability of these two pathogens to infect currently unreported hosts, inoculating central bearded dragons ( Pogona vitticeps ) with O. ophiodiicola and corn snakes ( Pantherophis guttatus ) with N. guarroi . We confirmed infection by documenting both clinical signs and histopathological evidence of fungal infection. Our reciprocity experiment resulted in 100% of corn snakes and 60% of bearded dragons developing infections with N. guarroi and O. ophiodiicola , respectively, demonstrating that these fungal pathogens have a broader host range than previously thought and that hosts with cryptic infections may play a role in pathogen translocation and transmission.

Applied and Environmental Microbiology↗

USGS2018

Understanding how microbiomes influence the systems they inhabit

Translating the ever-increasing wealth of information on microbiomes (environment, host, or built environment) to advance the understanding of system-level processes is proving to be an exceptional research challenge. One reason for this challenge is that relationships between characteristics of microbiomes and the system-level processes they influence are often evaluated in the absence of a robust conceptual framework and reported without elucidating the underlying causal mechanisms. The reliance on correlative approaches limits the potential to expand the inference of a single relationship to additional systems and advance the field. We propose that research focused on how microbiomes influence the systems they inhabit should work within a common framework and target known microbial processes that contribute to the system-level processes of interest. Here we identify three distinct categories of microbiome characteristics (microbial processes, microbial community properties, and microbial membership) and propose a framework to empirically link each of these categories to each other and the broader system level processes they affect. We posit that it is particularly important to distinguish microbial community properties that can be predicted from constituent taxa (community aggregated traits) from and those properties that are currently unable to be predicted from constituent taxa (emergent properties). Existing methods in microbial ecology can be applied to more explicitly elucidate properties within each of these categories and connect these three categories of microbial characteristics with each other. We view this proposed framework, gleaned from a breadth of research on environmental microbiomes and ecosystem processes, as a promising pathway with the potential to advance discovery and understanding across a broad range of microbiome science.

Nature Microbiology↗

USGS2002

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↗