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Acetylenotrophic and diazotrophic Bradyrhizobium sp. strain I71 from TCE-contaminated soils

Abstract Acetylene (C 2 H 2 ) is a molecule rarely found in nature, with very few known natural sources, but acetylenotrophic microorganisms can use acetylene as their primary carbon and energy source. As of 2018 there were 15 known strains of aerobic and anaerobic acetylenotrophs; however, we hypothesize there may yet be unrecognized diversity of acetylenotrophs in nature. This study expands the known diversity of acetylenotrophs by isolating the aerobic acetylenotroph, Bradyrhizobium sp. strain I71, from trichloroethylene (TCE)-contaminated soils. Strain I71 is a member of the class Alphaproteobacteria and exhibits acetylenotrophic and diazotrophic activities, the only two enzymatic reactions known to transform acetylene. This unique capability in the isolated strain may increase the genus’ economic impact beyond agriculture as acetylenotrophy is closely linked to bioremediation of chlorinated contaminants. Computational analyses indicate that the Bradyrhizobium sp. strain I71 genome contains 522 unique genes compared to close relatives. Moreover, applying a novel hidden Markov model of known acetylene hydratase (AH) enzymes identified a putative AH enzyme. Protein annotation with I-TASSER software predicted the AH from the microbe Syntrophotalea acetylenica as the closest structural and functional analog. Furthermore, the putative AH was flanked by horizontal gene transfer (HGT) elements, like that of AH in anaerobic acetylenotrophs, suggesting an unknown source of acetylene or acetylenic substrate in the environment that is selecting for the presence of AH. Importance The isolation of Bradyrhizobium strain I71 expands the distribution of acetylene-consuming microbes to include a group of economically important microorganisms. Members of Bradyrhizobium are well studied for their abilities to improve plant health and increase crop yields by providing bioavailable nitrogen. Additionally, acetylene-consuming microbes have been shown to work in tandem with other microbes to degrade soil contaminants. Based on genome, cultivation, and protein prediction analysis, the ability to consume acetylene is likely not widespread within the genus Bradyrhizobium . These findings suggest that the suite of phenotypic capabilities of strain I71 may be unique and make it a good candidate for further study in several research avenues.

California↗

Nitrate is a preferred electron acceptor for growth of freshwater selenate-respiring bacteria

An anaerobic, freshwater enrichment grew with either nitrate or selenate as an electron acceptor. With both ions present, nitrate reduction preceded selenate reduction. An isolate from the enrichment grew on either ion, but the presence of nitrate precluded the reduction of selenate. Stock cultures of denitrifiers grew anaerobically on nitrate but not on selenate.

Applied and Environmental Microbiology↗

Krumholzibacteriota and Deltaproteobacteria contain rare genetic potential to liberate carbon from monoaromatic compounds in subsurface coal seams

Biogenic methane in subsurface coal seam environments is produced by diverse consortia of microbes. Although this methane is useful for global energy security, it remains unclear which microbes can liberate carbon from the coal. Most of this carbon is relatively resistant to biodegradation, as it is contained within aromatic rings. Thus, to explore for coal-degrading taxa in the subsurface, this study reconstructed relevant metagenome-assembled genomes (MAGs) from coal seams by using a key genomic marker for the anaerobic degradation of monoaromatic compounds as a guide: the benzoyl-CoA reductase gene ( bcrABCD ). Three MAGs were identified with this genetic potential. The first represented a novel taxon from the Krumholzibacteriota phylum, which this study is the first to describe. This Krumholzibacteriota MAG contained a full set of genes for benzoyl-CoA dearomatization, in addition to other genes for anaerobic catabolism of monoaromatics. Analysis of Krumholzibacteriota MAGs from other environments revealed that this genetic potential may be common, and thus, Krumholzibacteriota may be important organisms for the liberation of recalcitrant carbon in a broad range of environments. Moreover, the assembly and characterization of two Syntrophorhabdus aromaticivorans MAGs from different continents and a Syntrophaceae sp. MAG implicate the Deltaproteobacteria class in coal seam monoaromatic degradation. Each of these taxa are potential rate-limiting organisms for subsurface coal-to-methane biodegradation. Their description here provides some understanding of their function within the coal seam microbiome and will help inform future efforts in coal bed methane stimulation, anoxic bioremediation of organic pollutants, and assessments of anoxic, subsurface carbon cycling and emissions.

mBio↗

Green River oil shale—concept of origin extended: An interdisciplinary problem being attacked from both ends

A much fuller understanding of the Green River oil shale and its organic chemistry will emerge when the geologists, paleontologists, organic chemists, biologists, paleolimnologists, and biogeochemists, who are now working on it, integrate their findings with those of the others. We know from the geology, paleontology, and paleolimnology that the biologic progenitors of the organic substance in the Green River oil shale could only have been microscopic algae, and other micro-organisms, that grew and accumulated in the central parts of large, shallow lakes that existed under a subtropical climate. The only nonlacustrine organic components were wind-blown, or water borne, pollens and waxy spores. These, however, made up a large and important part of the organic-rich sediment. The geology of the Green River Formation shows that as the algal and pollen-rich sediment was buried deeper and deeper, progressively more of its pore water and dissolved constituents were expressed. Static pressures may have reached as much as 210 kg cm −2 , and the ambient temperature rose, with depth, to somewhere within the range between 90° and 125° C. Beneath the ancient lakes a tectonically quiescent environment persisted for tens of millions of years after their organic sediments had been deeply buried. The organic material of the Green River oil shale can be divided into three fractions—a small bitumen fraction that is extractable with common organic solvents, a major fraction called koerogen that consists of insoluble pyrobitumens, and a somewhat smaller inert fraction that is neither soluble nor does it yield oil on pyrolysis. As all three fractions originated in the same algal, pollen-rich sediment, an explanation for their marked differences must be sought in their geochemical history or from a study of the modern analogues of their progenitors. The components of the bitumen fraction consisted of “biological markers” that were inherited from the Eocene plants and animals in which they originally formed. Diagenesis has changed these hydrogen-rich compounds, but not enough to obscure their provenance. Kerogen presumably became insoluble because its hydrogen-rich components polymerized. My speculation is that the inert fraction was derived from a polyphenolic substance produced in the original algal ooze by “non-enzymatic browning.” Only three Classes of non-marine algae need be considered as progenitors of the Green River oil shale; the Xanthophyceae, the Chlorophyceae, and the Cyanophyceae. Only the Cyanophyceae (the blue-green algae) meet the biologic and paleontologic requirements to have served as the dominant precursors of the Green River oil shale. Several other oil shales clearly were derived from the Xanthophyceae, specifically Botryococcus. The blue-green algal ooze now forming, and accumulating, in Mud Lake, Florida, has been studied biologically and chemically as a possible present-day analogue of the Green River oil shale precursor. In this small lake we have established the fact that a bacterial inhibitor is produced, which inhibits decay of the algae and thereby permits the accumulation of energy-rich organic compounds. We infer that a similar indigenous inhibitor must have acted in the Eocene lakes to permit them to become the huge energy sinks they were. Studies of the organic chemistry of living blue-green algae show that they contain appreciable percentages of fatty acids, hydrocarbons, and very large percentages of proteins. These promising, energy-rich compounds could serve as source materials for potential conversion into oil shale in the geologic future. Certain marine anaerobic bacteria convert fatty acids into aliphatic hydrocarbons. Fresh-water obligate anaerobes should be investigated to see if they also convert fatty acids into hydrocarbons. The part played by aquatic animals that live in, or on, freshwater algal ooze in synthesizing hydrocarbons has not been investigated, but deserves attention. Pollen grains, of course, must be considered an important precursor of hydrocarbons produced on pyrolysis. They contain far higher percentages of long chain hydrocarbons and alcohols than most plant materials. The major problem ahead is to account for the progressive hydrogenation and subsequent polymerization of the relatively oxygen-rich constituents of algae such as the polysaccharides, amino acids, ammo sugars, and fatty acids into the insoluble pyrobitumens that constitute, particularly, the kerogen fraction of the Green River oil shale.

Wyoming↗

Methanogenic calcite, 13C-depleted bivalve shells, and gas hydrate from a mud volcano offshore southern California

Methane and hydrogen sulfide vent from a cold seep above a shallowly buried methane hydrate in a mud volcano located 24 km offshore southern California in?? 800 m of water. Bivalves, authigenic calcite, and methane hydrate were recovered in a 2.1 m piston core. Aragonite shells of two bivalve species are unusually depleted in 13C (to -91??? ??13C), the most 13C-depleted shells of marine macrofauna yet discovered. Carbon isotopes for both living and dead specimens indicate that they used, in part, carbon derived from anaerobically oxidized methane to construct their shells. The ??13C values are highly variable, but most are within the range -12??? to -91???. This variability may be diagnostic for identifying cold-seep-hydrate systems in the geologic record. Authigenic calcite is abundant in the cores down to ???1.5 m subbottom, the top of the methane hydrate. The calcite is depleted in 13C (??13C = -46??? to -58???), indicating that carbon produced by anaerobically oxidized methane is the main source of the calcite. Methane sources include a geologic hydrocarbon reservoir from Miocene source rocks, and biogenic and thermogenic degradation of organic matter in basin sediments. Oxygen isotopes indicate that most calcite formed out of isotopic equilibrium with ambient bottom water, under the influence of gas hydrate dissociation and strong methane flux. High metal content in the mud volcano sediment indicates leaching of basement rocks by fluid circulating along an underlying fault, which also allows for a high flux of fossil methane. ?? 2006 Geological Society of America.

Geology↗

Arsenic and selenium in microbial metabolism

Arsenic and selenium are readily metabolized by prokaryotes, participating in a full range of metabolic functions including assimilation, methylation, detoxification, and anaerobic respiration. Arsenic speciation and mobility is affected by microbes through oxidation/reduction reactions as part of resistance and respiratory processes. A robust arsenic cycle has been demonstrated in diverse environments. Respiratory arsenate reductases, arsenic methyltransferases, and new components in arsenic resistance have been recently described. The requirement for selenium stems primarily from its incorporation into selenocysteine and its function in selenoenzymes. Selenium oxyanions can serve as an electron acceptor in anaerobic respiration, forming distinct nanoparticles of elemental selenium that may be enriched in (76)Se. The biogenesis of selenoproteins has been elucidated, and selenium methyltransferases and a respiratory selenate reductase have also been described. This review highlights recent advances in ecology, biochemistry, and molecular biology and provides a prelude to the impact of genomics studies.

Annual Review of Microbiology↗

Microbial sulfate reduction and metal attenuation in pH 4 acid mine water

Sediments recovered from the flooded mine workings of the Penn Mine, a Cu-Zn mine abandoned since the early 1960s, were cultured for anaerobic bacteria over a range of pH (4.0 to 7.5). The molecular biology of sediments and cultures was studied to determine whether sulfate-reducing bacteria (SRB) were active in moderately acidic conditions present in the underground mine workings. Here we document multiple, independent analyses and show evidence that sulfate reduction and associated metal attenuation are occurring in the pH-4 mine environment. Water-chemistry analyses of the mine water reveal: (1) preferential complexation and precipitation by H2S of Cu and Cd, relative to Zn; (2) stable isotope ratios of 34S/32S and 18O/16O in dissolved SO4 that are 2-3 ??? heavier in the mine water, relative to those in surface waters; (3) reduction/oxidation conditions and dissolved gas concentrations consistent with conditions to support anaerobic processes such as sulfate reduction. Scanning electron microscope (SEM) analyses of sediment show 1.5-micrometer, spherical ZnS precipitates. Phospholipid fatty acid (PLFA) and denaturing gradient gel electrophoresis (DGGE) analyses of Penn Mine sediment show a high biomass level with a moderately diverse community structure composed primarily of iron- and sulfate-reducing bacteria. Cultures of sediment from the mine produced dissolved sulfide at pH values near 7 and near 4, forming precipitates of either iron sulfide or elemental sulfur. DGGE coupled with sequence and phylogenetic analysis of 16S rDNA gene segments showed populations of Desulfosporosinus and Desulfitobacterium in Penn Mine sediment and laboratory cultures. ?? 2007 Church et al; licensee BioMed Central Ltd.

Geochemical Transactions↗

Paleoenvironment of the New Albany Shale Group ( Devonian- Mississippian) of Illinois

The distribution of lithofacies in the New Albany Shale Group of Illinois was determined by wave energy, bottom oxygenation, and bottom topography in a deep water stratified anoxic basin. A transect from the margin to the center of the Illinois Basin reveals a complete transition from high energy, aerobic, shallow-water environments to quiet, anaerobic, deep-water environments. Shallow areas at the margin of the basin are characterized by rapid facies transitions over short distances. High energy, very shallow conditions are recorded by oolitic-skeletal grainstones and packstones with abundant brachiopods, crinoids, trilobites, and other calcified marine invertebrates. Bioturbation did not destroy primary sedimentary structures in these facies. Offshore, less agitated areas are represented by highly bioturbated carbonate wackestones, argillaceous quartz siltstones, and greenish-gray mudstones. Calcified invertebrates are generally rare in these facies, indicating deposition in dysaerobic conditions. Basinward, slope areas are characterized by olive-gray to black, weakly bioturbated shales commonly interbedded with thickly laminated black shales. Trace fossils, including Zoophycos, Chondrites , and Planolites , are abundant along the bases of the olive-gray beds. In areas where the anaerobic/dysaerobic boundary intersected the bottom slope, slight fluctuations of the position of the boundary resulted in thin interbedding of olive-gray and black shales and laterally persistent interfingering of the two lithologies. Anaerobic conditions prevailed during most of New Albany time in the deepest areas of the basin, and finely laminated, undisturbed, pelagic black shales were deposited.

Journal of Sedimentary Petrology↗

The ecology of methane in streams and rivers: Patterns, controls, and global significance

Streams and rivers can substantially modify organic carbon (OC) inputs from terrestrial landscapes, and much of this processing is the result of microbial respiration. While carbon dioxide (CO 2 ) is the major end-product of ecosystem respiration, methane (CH 4 ) is also present in many fluvial environments even though methanogenesis typically requires anoxic conditions that may be scarce in these systems. Given recent recognition of the pervasiveness of this greenhouse gas in streams and rivers, we synthesized existing research and data to identify patterns and drivers of CH 4 , knowledge gaps, and research opportunities. This included examining the history of lotic CH 4 research, creating a database of concentrations and fluxes (MethDB) to generate a global-scale estimate of fluvial CH 4 efflux, and developing a conceptual framework and using this framework to consider how human activities may modify fluvial CH 4 dynamics. Current understanding of CH 4 in streams and rivers has been strongly influenced by goals of understanding OC processing and quantifying the contribution of CH 4 to ecosystem C fluxes. Less effort has been directed towards investigating processes that dictate in situ CH 4 production and loss. CH 4 makes a meager contribution to watershed or landscape C budgets, but streams and rivers are often significant CH 4 sources to the atmosphere across these same spatial extents. Most fluvial systems are supersaturated with CH 4 and we estimate an annual global emission of 26.8 Tg CH 4 , equivalent to ~15-40% of wetland and lake effluxes, respectively. Less clear is the role of CH 4 oxidation, methanogenesis, and total anaerobic respiration to whole ecosystem production and respiration. Controls on CH 4 generation and persistence can be viewed in terms of proximate controls that influence methanogenesis (organic matter, temperature, alternative electron acceptors, nutrients) and distal geomorphic and hydrologic drivers. Multiple controls combined with its extreme redox status and low solubility result in high spatial and temporal variance of CH 4 in fluvial environments, which presents a substantial challenge for understanding its larger-scale dynamics. Further understanding of CH 4 production and consumption, anaerobic metabolism, and ecosystem energetics in streams and rivers can be achieved through more directed studies and comparison with knowledge from terrestrial, wetland, and aquatic disciplines.

Ecological Monographs↗

Relating hyporheic fluxes, residence times, and redox-sensitive biogeochemical processes upstream of beaver dams

Abstract. Small dams enhance the development of patchy microenvironments along stream corridors by trapping sediment and creating complex streambed morphologies. This patchiness drives intricate hyporheic flux patterns that govern the exchange of O 2 and redox-sensitive solutes between the water column and the stream bed. We used multiple tracer techniques, naturally occurring and injected, to evaluate hyporheic flow dynamics and associated biogeochemical cycling and microbial reactivity around 2 beaver dams in Wyoming (USA). High-resolution fiber-optic distributed temperature sensing was used to collect temperature data over 9 vertical streambed profiles and to generate comprehensive vertical flux maps using 1-dimensional (1-D) heat-transport modeling. Coincident with these locations, vertical profiles of hyporheic water were collected every week and analyzed for dissolved O 2 , pH, dissolved organic C, and several conservative and redox-sensitive solutes. In addition, hyporheic and net stream aerobic microbial reactivity were analyzed with a constant-rate injection of the biologically sensitive resazurin (Raz) smart tracer. The combined results revealed a heterogeneous system with rates of downwelling hyporheic flow organized by morphologic unit and tightly coupled to the redox conditions of the subsurface. Principal component analysis was used to summarize the variability of all redox-sensitive species, and results indicated that hyporheic water varied from oxic-stream-like to anoxic-reduced in direct response to the hydrodynamic conditions and associated residence times. The anaerobic transition threshold predicted by the mean O 2 Damko ¨hler number seemed to overestimate the actual transition as indicated by multiple secondary electron acceptors, illustrating the gradient nature of anaerobic transition. Temporal flux variability in low-flux morphologies generated a much greater range in hyporheic redox conditions compared to high-flux zones, and chemical responses to changing flux rates were consistent with those predicted from the empirical relationship between redox condition and residence time. The Raz tracer revealed that hyporheic flow paths have strong net aerobic respiration, particularly at higher residence time, but this reactive exchange did not affect the net stream signal at the reach scale.

Freshwater Science↗

Sulfur and oxygen isotopes in barite deposits of the western Brooks Range, Alaska, and implications for the origin of the Red Dog massive sulfide deposits

Sulfur and oxygen isotope analyses have been obtained for barite samples from the giant stratiform sulfide barite deposits at Red Dog in the western Brooks Range of Alaska, from stratiform barite deposits elsewhere in the Red Dog district, and from stratiform and vein and breccia barite occurrences in the central Brooks Range. Twelve of the 15 deposits studied lie within middle to Upper Mississippian black shale and chert units. The data reveal two different patterns on ?? 34S versus ??18O plots. The first, which is best illustrated by the barite deposit at Anarraaq, shows linear trends with slopes that vary with barite texture. For most samples, ??34S and ??18O values are both higher than the values characteristic of Mississippian marine sulfate. The second pattern, which is evident at the Red Dog deposits, shows no correlation between ??34S and ??18. In most samples, ??18O is below the value for Mississippian marine sulfate. Comparisons with sulfate in modern marine environments suggest a possible model for the mineralizing process. Anarraaq-type barite formed at sea-floor vents where ascending fluids carrying barium and methane encountered sulfate-bearing pore waters or bottom waters. Barite deposition was accompanied by the reduction of sulfate to H2S by means of microbially mediated anaerobic methane oxidation. Red Dog-type barite was formed in a manner similar to Anarraaq-type barite but was over-printed by a massive sulfide-forming event. Red Dog sulfides precipitated where metal-bearing hydrothermal fluids encountered pore waters that had been charged with H2S by anaerobic methane oxidation. Textural and isotopic evidence indicates that the sulfide bodies grew by consuming the available H2S and then by reductively dissolving barite. Dissolution of barite caused barium to be released to higher stratigraphic levels where it was reprecipitated on encountering sulfate. Isotopic evidence is pre sented for a link between methane venting and barite formation and raises the possibility that the coexistence of barite and sulfide at Red Dog, and the occurrence elsewhere in the district of barite-only and sulfide-only deposits, can be explained by a spectrum of vent types in the Mississippian basin analogous to the spectrum that is observed today along the modern continental margins. Authigenic barite formed at some but not all methane seeps, perhaps owing to differences in the barium content of vent fluids, differences in the relative proportion of aqueous fluid and gas emanating from vents, or differences in sulfate availability in local bottom waters. Some barite-forming seeps were later replaced by sulfides (Red Dog deposits) whereas others were not (e.g., Anarraaq barite horizon, Gull Creek, Moil). At sulfide occurrences where there is little evidence of preexisting barite (e.g., Anarraaq, Wulik, Suds), methane venting is indicated by fossils suggestive of chemosynthetic fauna. Mammiform sedimentary structures that are widespread in black chert at the top of the Kuna Formation may represent seeps that supported neither authigenic mineral formation nor chemosynthetic megafauna. ?? 2004 by Economic Geology.

Economic Geology↗

Survival of bacterial indicators and the functional diversity of native microbial communities in the Floridan aquifer system, south Florida

The Upper Floridan aquifer in the southern region of Florida is a multi-use, regional scale aquifer that is used as a potable water source and as a repository for passively recharged untreated surface waters, and injected treated surface water and wastewater, industrial wastes, including those which contain greenhouse gases (for example, carbon dioxide). The presence of confined zones within the Floridan aquifer that range in salinity from fresh to brackish allow regulatory agencies to permit the injection of these different types of product waters into specific zones without detrimental effects to humans and terrestrial and aquatic ecosystems. The type of recharge that has received the most regulatory attention in south Florida is aquifer storage and recovery (ASR). The treated water, prior to injection and during recovery, must meet primary and secondary drinking water standards. The primary microbiology drinking water standard is total coliforms, which have been shown to be difficult to inactivate below the regulatory standard during the treatment process at some ASR facilities. The inefficient inactivation of this group of indicator bacteria permits their direct injection into the storage zones of the Floridan aquifer. Prior to this study, the inactivation rates for any member of the total coliform group during exposure to native geochemical conditions in groundwater from any zone of the Floridan aquifer had not been derived. Aboveground flow through mesocosms and diffusion chambers were used to quantify the inactivation rates of two bacterial indicators, Escherichia coli and Pseudomonas aeruginosa, during exposure to groundwater from six wells. These wells collect water from two ASR storage zones: the Upper Floridan aquifer (UFA) and Avon Park Permeable Zone (APPZ). Both bacterial strains followed a biphasic inactivation model. The E. coli populations had slower inactivation rates in the UFA (range: 0.217–0.628 per hour (h -1 )) during the first phase of the model than when exposed to groundwater from the APPZ (range: 0.540–0.684 h -1 ). The inactivation rates for the first phase of the models for P. aeruginosa were not significantly different between the UFA (range: 0.144–0.770 h -1 ) and APPZ (range: 0.159–0.772 h -1 ) aquifer zones. The inactivation rates for the second phase of the model for this P. aeruginosa were also similar between UFA (range: 0.003–0.008 h -1 ) and APPZ (0.004–0.005 h -1 ) zones, although significantly slower than the model’s first phase rates for this bacterial species. Geochemical data were used to determine which dissimilatory biogeochemical reactions were most likely to occur under the native conditions in the UFA and APPZ zones using thermodynamics principles to calculate free energy yields and other cell-related energetics data. The biogeochemical processes of acetotrophic and hydrogenotrophic sulfate reduction, methanogenesis and anaerobic oxidation of methane dominated in all six groundwater sites. A high throughput DNA microarray sequencing technology was used to characterize the diversity in the native aquifer bacterial communities (bacteria and archaea) and assign putative physiological capabilities to the members of those communities. The bacterial communities in both zones of the aquifer were shown to possess the capabilities for primary and secondary fermentation, acetogenesis, methanogenesis, anaerobic methane oxidation, syntrophy with methanogens, ammonification, and sulfate reduction. The data from this study provide the first determination of bacterial indicator survival during exposure to native geochemical conditions of the Floridan aquifer in south Florida. Additionally, the energetics and functional bacterial diversity characterizations are the first descriptions of native bacterial communities in this region of the Floridan aquifer and reveal how these communities persist under such extreme conditions. Collectively, these types of data can be used to develop and refine groundwater models.

Florida↗

Surface-Water Hydrology and Quality at the Pike Hill Superfund Site, Corinth, Vermont, October 2004 to December 2005

The hydrology and quality of surface water in and around the Pike Hill Brook watershed, in Corinth, Vermont, was studied from October 2004 to December 2005 by the U.S. Geological Survey in cooperation with the U.S. Environmental Protection Agency (USEPA). Pike Hill was mined intermittently for copper from 1847 to 1919 and the site is known to be contributing trace elements and acidity to Pike Hill Brook and an unnamed tributary to Cookville Brook. The site has been listed as a Superfund site since 2004. Streamflow, specific conductance, pH, and water temperature were measured continuously and monthly at three sites on Pike Hill Brook to determine the variation in these parameters over an annual cycle. Synoptic water-quality sampling was done at 10 stream sites in October 2004, April 2005, and June 2005 and at 13 stream sites in August 2005 to characterize the quality of surface water in the watershed on a seasonal and spatial basis, as well as to assess the effects of wetlands on water quality. Samples for analysis of benthic macroinvertebrate populations were collected at 11 stream sites in August 2005. Water samples were analyzed for 5 major ions and 32 trace elements. Concentrations of trace elements at sites in the Pike Hill Brook watershed exceeded USEPA National Recommended Water Quality Criteria acute and chronic toxicity standards for aluminum, iron, cadmium, copper, and zinc. Concentrations of copper exceeded the chronic criteria in an unnamed tributary to Cookville Brook in one sample. Concentrations of sulfate, calcium, aluminum, iron, cadmium, copper, and zinc decreased with distance from a site directly downstream from the mine (site 1), as a result of dilution and through sorption and precipitation of the trace elements. Maximum concentrations of aluminum, iron, cadmium, copper, and zinc were observed during spring snowmelt. Concentrations of sulfate, calcium, cadmium, copper, and zinc, and instantaneous loads of calcium and aluminum were statistically different (p<0.05) among the three continuously monitored sites (sites 1, 4, and 5). Instantaneous loads of aluminum, iron, and copper decreased by one to three orders of magnitude from site 1 to a site 1.1 mi downstream (site 4). Instantaneous loads of sulfate were similar between sites 1, 4, and at a site 3 mi downstream (site 5). Instantaneous loads of cadmium and zinc were similar between sites 1 and 4, and loads of iron and copper were similar between sites 4 and 5. Loads of chemical constituents were compared at site 1 (closest to the mine waste piles) and site 5 (near the mouth of Pike Hill Brook and below a majority of the wetlands). Annually, the loads of dissolved cadmium and zinc at site 1 were about five times greater than loads at site 5, and the load of dissolved copper at site 1 was about 17 times greater than at site 5. The ratio of loads for dissolved cadmium, copper, and zinc to total cadmium, copper, and zinc at site 1 was about 1. Samples collected in Pike Hill Brook upstream and downstream from the wetlands during low flows in August 2005 showed that oxidation of ferrous iron and precipitation of iron-hydroxides were probably not affecting trace metals in the wetlands through sorption; however, a significant portion of the iron entering the wetlands was in particulate form and may have transported sorbed copper and other trace metals. Thus, aerobic activity in the wetlands was probably not affecting metal cycling in the watershed. Concentrations and loads of sulfate may be unlikely to define unequivocally the role of the wetlands with regard to anaerobic bacterial sulfate reduction; however, bacterial sulfate removal may have affected loads of sulfate. Loads of copper increased downstream from the wetlands and may reflect the reductive dissolution of ferric hydroxide particulates in anaerobic parts of the wetlands.Concentrations of dissolved iron increased downstream from the wetlands. The most apparent effects on the macroinvertebr

Scientific Investigations Report↗

Fate and transport of petroleum hydrocarbons in the subsurface near Cass Lake, Minnesota

The U.S. Geological Survey (USGS) investigated the natural attenuation of subsurface petroleum hydrocarbons leaked over an unknown number of years from an oil pipeline under the Enbridge Energy Limited Partnership South Cass Lake Pumping Station, in Cass Lake, Minnesota. Three weeks of field work conducted between May 2007 and July 2008 delineated a dissolved plume of aromatic hydrocarbons and characterized the biodegradation processes of the petroleum. Field activities included installing monitoring wells, collecting sediment cores, sampling water from wells, and measuring water-table elevations. Geochemical measurements included concentrations of constituents in both spilled and pipeline oil, dissolved alkylbenzenes and redox constituents, sediment bioavailable iron, and aquifer microbial populations. Groundwater in this area flows east-southeast at approximately 26 meters per year. Results from the oil analyses indicate a high degree of biodegradation, characterized by nearly complete absence of n-alkanes. Cass Lake oil samples were more degraded than two oil samples collected in 2008 from the similarly contaminated USGS Bemidji, Minnesota, research site 40 kilometers away. Based on 19 ratios developed for comparing oil sources, the conclusion is that the oils at the two sites appear to be from the same hydrocarbon source. In the Cass Lake groundwater plume, benzene concentrations decrease by three orders of magnitude within 150 meters (m) downgradient from the oil body floating on the water table (between well MW-10 and USGS-4 well nest). The depths of the highest benzene concentrations increase with distance downgradient from the oil, a condition typical of plumes in shallow, unconfined aquifers. Background groundwater, which is nearly saturated with oxygen, becomes almost entirely anaerobic in the plume. As at the Bemidji site, the most important biodegradation processes are anaerobic and dominated by iron reduction. The similarity between the Cass Lake and Bemidji benzene degradation rates, redox conditions, and aquifer material all support a hypothesis that the Cass Lake plume, like the Bemidji plume, is decades old. As concentrations of alkylbenzenes in the oil decrease over time, the benzene concentrations in the groundwater plume will also decrease and the plume is expected to shrink. The Fox Creek wetland, about 250 m south of the Cass Lake site, is the nearest receptor to the south.

Scientific Investigations Report↗

Limnology of Taylor Creek impoundment with reference to other water bodies in Upper St. Johns River Basin, Florida

Taylor Creek Impoundment was constructed on the western side of the upper St. Johns River basin as part of a plan for flood control and water regulation. The impoundment, which has a surface area of about 4,000 acres, was initially filled late in 1969. Water of relatively poor quality was observed in the impoundment during its first three years of its existence (1970-72). The depth of the impoundment is sufficient to allow thermal stratification, and a thermocline usually develops at depths of 8 to 10 feet. During 1970-72 the hypolimnion remained anaerobic for more than half the year. The hypolimnion also accumulated high concentrations of phosphorus, ammonia-nitrogen, carbon dioxide, ferrous iron, hydrogen sulfide and other substances. The poor water quality is attributed to the decomposition of flooded vegetation, decomposition of soil organic matter, and heavy growths of phytoplankton and duckweed stimulated by an abundant supply of nutrients. Flushing of the impoundment and depletion of leachable nutrients and soil organic matter have led to an improvement in water quality since 1972. During 1973 and 1974 the depth to the top of the anaerobic zone increased to more than 12 feet and by 1974 the period of anaerobiosis decreased to less than 2 months out of 12. Phosphorus concentration after the fall overturn decreased more than 50 percent between 1970 and 1974. The reduction in biochemical oxygen demand during this period suggests a decrease in primary productivity. Water released from the impoundment during the period 1969-75 was similar in quality to nearby Wolf Creek and Jane Green Creek. Of 21 physical, organic, and inorganic constituents, only ammonia-nitrogen was significantly higher in releases from the impoundment than it was in the natural streams. Dissolved oxygen was higher in water released from the impoundment than in the natural streams and dissolved solids concentration were lower. Large releases from the impoundment may, under certain conditions, produce velocities great enough to resuspend bottom sediments several miles downstream at a point where Taylor Creek flows into Lake Poinsett.

Florida↗

Tests for injecting, storing, and recovering freshwater in a saline artesian aquifer, Lee County, Florida

An investigation was made of the suitability of a saline, artesian limestone aquifer for the injection, storage, and recovery of freshwater from the Caloosahatchee River. The tests were conducted on a well tapping a leaky artesian system that has a transmissivity of 800 square feet per day, a storage of 1 x 10-4, and a leakance of 0.01 per day. The specific capacity of the injection well was increased through acidizing and was decreased as a result of well clogging during injection. Three injection tests were made wherein the amounts of freshwater injected, the storage duration, and the quality of water injected varied. Analysis of the test data showed that freshwater recoverability ranged from 9.7 to 38.7 percent of the total injected. Differences were attributed principally to differences in the quality of water injected and storage duration. Repeated injection-recovery cycles probably would result in greater recoverability. Head buildup, nearly 200 feet in one test, was a prime problem related chiefly to clogging from suspended material in the injected water and to bacterial growth at the wellbore-limestone interface. Regular backflushing was required. Total head buildup decreased as a result of acidizing the injection well. No coliforms or fecal streptococcus were noted in the recovered water. Growth of anaerobic bacteria occurred. Changes in the quality of the recovered water included decreases in concentration of dissolved organic carbon by as much as 15 mg/L (milligrams per liter), organic nitrogen by as much as 0.80 mg/L, and nitrate by as much as 0.50 mg/L. Increases were noted in ammonia by 0.40 mg/L, and iron by as much as 0.60 mg/L. These changes are consistent with the presence of an anaerobic bacterial ecosystem.

Florida↗

Effects of two stormwater management methods on the quality of water in the upper Biscayne aquifer at two commercial areas in Dade County, Florida

This study is part of a continued effort to assess the effects of urban stormwater recharge on the water quality of the Biscayne aquifer in southeast Florida. In this report, the water-quality effects on shallow ground water resulting from stormwater disposal by exfiltration trench and grassy swale were investigated at two small commercial areas in Dade County, Florida. One study area (airport) was located near the Miami International Airport and had a drainage area of about 10 acres overlying a sandy soil; the other study area (free zone) was located at the Miami International Free Trade Zone and had a drainage area of about 20 acres overlying limestone. The monitoring design for each study area consisted of seven sites and included water-quality sampling of the stormwater in the catch basin of the exfiltration trench, ground water from two wells 1 foot from the trench (trench wells), two wells 20 feet from the trench, and ground water from two wells at the swale from April 1985 through May 1986. Eleven water-quality variables (target variables) commonly found in high levels in urban stormwater runoff were used as tracers to estimate possible changes in ground-water quality that may have been caused by stormwater recharge. Comparison of the distribution of target variables indicated that the concentrations tended to be greater in the stormwater in the exfiltration trench than in water from the two wells 1 foot from the trench at both study areas. The concentration difference for several target variables was statistically significant at the 5-percent level. Lead, for example, had median concentrations of 23 and 4 micrograms per liter, respectively, in stormwater and water from the two trench wells at the airport study area, and 38 and 2 micrograms per liter, respectively, in stormwater and groundwater at the free zone. Similar reductions in concentrations between stormwater and water from the two trench wells were indicated for zinc at both study areas and also for nitrogen, phosphorus, and organic content at the free zone. This trend suggested that the exfiltration trench at both study areas may function as a partial trap for some chemical substances present in stormwater. A comparison of the distribution of the 11 target variables and major ionic composition in water from the two trench wells and the two wells 20 feet from the trench did not indicate a notable horizontal stratification at either study area. A vertical difference between 10 and 15 feet, however, was indicated at the free zone with major ions in greater concentrations at 15 feet. The vertical variability in groundwater near the trench at the free zone may have been the result of stormwater dilution in the upper (10-foot) zone. The ground-water quality at the swale was quite dissimilar to that near the exfiltration trench at both the airport and free zone study areas. Data indicated that the groundwater environment at both sales was anaerobic as evidenced by abundant ammonia nitrogen and iron and trace levels of sulfate. Anaerobic conditions at the swale may have been the result of poor drainage and high organic content of soils. Significant biochemical cycling in the ground water at the swales precluded any assessment of quality effects that may result from storm-water infiltration.

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Hydrology, vegetation, and soils of four north Florida River flood plains with an evaluation of state and federal wetland determinations

A study of hydrologic conditions, vegetation, and soils was made in wetland forests of four north Florida streams from 1987 to 1990. The study was conducted by the U.S. Geological Survey in cooperation with the Florida Department of Environmental Regulation to support State and Federal efforts to improve wetland delineation methodology in flood plains. Plant communities and soils were described and related to topographic position and long-term hydrologic conditions at 10 study plots located on 4 streams. Detailed appendixes give average duration, frequency, and depth of flooding; canopy, subcanopy, and ground-cover vegetation; and taxonomic classification, series, and profile descriptions of soils for each plot. Topographic relief, range in stage, and depth of flooding were greatest on the alluvial flood plain of the Ochlockonee River, the largest of the four streams. Soils were silty in the lower elevations of the flood plain, and tree communities were distinctly different in each topographic zone. The Aucilla River flood plain was dominated by levees and terraces with very few depressions or low backwater areas. Oaks dominated the canopy of both lower and upper terraces of the Aucilla flood plain. Telogia Creek is a blackwater stream that is a major tributary of the Ochlockonee River. Its low, wet flood plain was dominated by Wyssa ogeche (Ogeechee tupelo) trees, had soils with mucky horizons, and was inundated by frequent floods of very short duration. The St. Marks River, a spring-fed stream with high base flow, had the least topographic relief and lowest range in stage of the four streams. St. Marks soils had a higher clay content than the other streams, and limestone bedrock was relatively close to the surface. Wetland determinations of the study plots based on State and Federal regulatory criteria were evaluated. Most State and Federal wetland determinations are based primarily on vegetation and soil characteristics because hydrologic records are usually not available. In this study, plots were located near long-term gaging stations, thus wetland determinations based on plant and soil characteristics could be evaluated at sites where long-term hydrologic conditions were known. Inconsistencies among hydrology, vegetation, and soil determinations were greatest on levee communities of the Ochlockonee and Aucilla River flood plains. Duration of average annual longest flood was almost 2 weeks for both plots. The wetland species list currently used (1991) by the State lacks many ground-cover species common to forested flood plains of north Florida rivers. There were 102 ground-cover species considered upland plants by the State that were present on the nine annually flooded plots of this study. Among them were 34 species that grew in areas continuously flooded for an average of 5 weeks or more each year. Common flood-plain species considered upland plants by the State were: Hypoxis leptocarpa (yellow star-grass), and two woody vines, Brunnichia ovata (ladies' eardrops) and Campsis radican s (trumpet-creeper), which were common in areas flooded continuously for 6 to 9 weeks a year; Sebastiania fruticosa (Sebastian-bush), Chasmanthium laxum (spikegrass), and Panicum dichotomum (panic grass), which typically grew in areas flooded an average of 2 to 3 weeks or more per year; Vitis rotundifolia (muscadine) and Toxicodendron radicans (poison-ivy), usually occurring in areas flooded an average of 1 to 2 weeks a year; and Quercus virginiana (live oak) present most often in areas flooded approximately 1 week a year. Federal wetland regulations (1989) limited wetland jurisdiction to only those areas that are inundated or saturated during the growing season. However, year-round hydrologic records were chosen in this report to describe the influence of hydrology on vegetation, because saturation, inundation, or flowing water can have a variety of both beneficial and adverse effects on flood-plain vegetation at any time of the year. These effects can occur because: (1) Soil temperatures in north Florida are probably high enough in winter for anaerobic conditions to develop in saturated soils. (2) Many plants in the flood plains of north Florida are active in the nongrowing season and might be adapted to anaerobic conditions in winter. (3) Other effects of standing water, such as decreased light penetration, prevention of seed germination, and protection of seeds from herbivores; and effects of flowing water, such as scouring and deposition of sediments, seed dispersal, and mechanical injury, can occur any time of year.

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