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368 records · Page 21Linked to original sources

Microbial diversity in a hydrocarbon- and chlorinated-solvent- contaminated aquifer undergoing intrinsic bioremediation

A culture-independent molecular phylogenetic approach was used to survey constituents of microbial communities associated with an aquifer contaminated with hydrocarbons (mainly jet fuel) and chlorinated solvents undergoing intrinsic bioremediation. Samples were obtained from three redox zones: methanogenic, methanogenic-sulfate reducing, and iron or sulfate reducing. Small-subunit rRNA genes were amplified directly from aquifer material DNA by PCR with universally conserved or Bacteria - or Archaea -specific primers and were cloned. A total of 812 clones were screened by restriction fragment length polymorphisms (RFLP), approximately 50% of which were unique. All RFLP types that occurred more than once in the libraries, as well as many of the unique types, were sequenced. A total of 104 (94 bacterial and 10 archaeal) sequence types were determined. Of the 94 bacterial sequence types, 10 have no phylogenetic association with known taxonomic divisions and are phylogenetically grouped in six novel division level groups (candidate divisions WS1 to WS6); 21 belong to four recently described candidate divisions with no cultivated representatives (OP5, OP8, OP10, and OP11); and 63 are phylogenetically associated with 10 well-recognized divisions. The physiology of two particularly abundant sequence types obtained from the methanogenic zone could be inferred from their phylogenetic association with groups of microorganisms with a consistent phenotype. One of these sequence types is associated with the genus Syntrophus ; Syntrophus spp. produce energy from the anaerobic oxidation of organic acids, with the production of acetate and hydrogen. The organism represented by the other sequence type is closely related to Methanosaeta spp., which are known to be capable of energy generation only through aceticlastic methanogenesis. We hypothesize, therefore, that the terminal step of hydrocarbon degradation in the methanogenic zone of the aquifer is aceticlastic methanogenesis and that the microorganisms represented by these two sequence types occur in syntrophic association.

Applied and Environmental Microbiology

Elucidation of taste- and odor-producing bacteria and toxigenic cyanobacteria in a Midwestern drinking water supply reservoir by shotgun metagenomics analysis

While commonplace in clinical settings, DNA-based assays for identification or enumeration of drinking water pathogens and other biological contaminants remain widely unadopted by the monitoring community. In this study, shotgun metagenomics was used to identify taste-and-odor producers and toxin-producing cyanobacteria over a 2-year period in a drinking water reservoir. The sequencing data implicated several cyanobacteria, including Anabaena spp ., Microcystis spp ., and an unresolved member of the order Oscillatoriales as the likely principal producers of geosmin, microcystin, and 2-methylisoborneol (MIB), respectively. To further demonstrate this, quantitative PCR (qPCR) assays targeting geosmin-producing Anabaena and microcystin-producing Microcystis were utilized, and these data were fitted using generalized linear models and compared with routine monitoring data, including microscopic cell counts, sonde-based physicochemical analyses, and assays of all inorganic and organic nitrogen and phosphorus forms and fractions. The qPCR assays explained the greatest variation in observed geosmin (adjusted R 2 = 0.71) and microcystin (adjusted R 2 = 0.84) concentrations over the study period, highlighting their potential for routine monitoring applications. The origin of the monoterpene cyclase required for MIB biosynthesis was putatively linked to a periphytic cyanobacterial mat attached to the concrete drinking water inflow structure. We conclude that shotgun metagenomics can be used to identify microbial agents involved in water quality deterioration and to guide PCR assay selection or design for routine monitoring purposes. Finally, we offer estimates of microbial diversity and metagenomic coverage of our data sets for reference to others wishing to apply shotgun metagenomics to other lacustrine systems.

Kansas

Relationship between oxidative stress, pathology, and behavioral signs of lead poisoning in mallards

Some of the adverse effects of lead (Pb) may be associated with oxidative damage of lipids, proteins, or DNA. In a previous study a linkage was observed between the susceptibilities of waterfowl species to Pb poisoning with oxidative stress . To investigate this relationship among the individuals of a single species, for 3 wk 4 groups of 72 mallards were fed diets containing high or low levels of vitamin E (20 or 220 Ul/kg) and high or low levels of Pb (0 or 2 g/kg). During the first week of Pb exposure, mallards developed hemolytic anemia, and during the second week, signs of neurological impairment. Histological findings in the Pb-exposed mallards were hemosiderosis, demyelinization of sciatic and brachial nerves, and tumefaction of renal tubular epithelium with the presence of intranuclear inclusion bodies. Lipid peroxidation increased with Pb exposure in blood, liver, bile, and brain, but decreased in nerves. Glutathione (GSH) increased with Pb exposure in liver and bile, and its oxidized/reduced ratio only increased in bile. Pb exposure inhibited GSH peroxidase activity (GPX) in plasma, liver, and brain, and decreased protein thiols (PSH) in blood and liver. Vitamin E resulted in significantly lower lipid peroxidation in nerves of control birds relative to unsupplemented controls, but did not alleviate any sign of lead posioning. Pb-induced pathological changes associated with hepatic and nervous functions were significantly correlated with lower GPX activity and PSH concentrations in these tissues rather than lipid peroxidation. Data suggest that inhibition of antioxidant enzymes and interaction with sulfhydryl groups of proteins may play a more important role in Pb poisoning of waterfowl than lipid peroxidation.

Journal of Toxicology and Environmental Health, Pa

Genomics of Arctic cod

The Arctic cod (Boreogadus saida) is an abundant marine fish that plays a vital role in the marine food web. To better understand the population genetic structure and the role of natural selection acting on the maternally-inherited mitochondrial genome (mitogenome), a molecule often associated with adaptations to temperature, we analyzed genetic data collected from 11 biparentally-inherited nuclear microsatellite DNA loci and nucleotide sequence data from from the mitochondrial DNA (mtDNA) cytochrome b (cytb) gene and, for a subset of individuals, the entire mitogenome. In addition, due to potential of species misidentification with morphologically similar Polar cod (Arctogadus glacialis), we used ddRAD-Seq data to determine the level of divergence between species and identify species-specific markers. Based on the findings presented here, Arctic cod across the Pacific Arctic (Bering, Chukchi, and Beaufort Seas) comprise a single panmictic population with high genetic diversity compared to other gadids. High genetic diversity was indicated across all 13 protein-coding genes in the mitogenome. In addition, we found moderate levels of genetic diversity in the nuclear microsatellite loci, with highest diversity found in the Chukchi Sea. Our analyses of markers from both marker classes (nuclear microsatellite fragment data and mtDNA cytb sequence data) failed to uncover a signal of microgeographic genetic structure within Arctic cod across the three regions, within the Alaskan Beaufort Sea, or between near-shore or offshore habitats. Further, data from a subset of mitogenomes revealed no genetic differentiation between Bering, Chukchi, and Beaufort seas populations for Arctic cod, Saffron cod (Eleginus gracilis), or Walleye pollock (Gadus chalcogrammus). However, we uncovered significant differences in the distribution of microsatellite alleles between the southern Chukchi and central and eastern Beaufort Sea samples of Arctic cod. Finally, using ddRAD-Seq data, we identified species-specific markers and in conjunction with mitogenome data, identified an Arctic cod x Polar cod hybrid in western Canadian Beaufort Sea. Overall, the lack of genetic structure among Arctic cod within the Bering, Chukchi and Beaufort seas of Alaska is concordant with the absence of geographic barriers to dispersal and typical among marine fishes. Arctic cod may exhibit a genetic pattern of isolation-by-distance, whereby populations in closer geographic proximity are more genetically similar than more distant populations. As this signal is only found between our two fartherest localities, data from populations elsewhere in the species’ global range are needed to determine if this is a general characteristic. Further, tests for selection suggested a limited role for natural selection acting on the mitochondrial genome of Arctic cod, but do not exclude the possibility of selection on genes involved in nuclear-mitogenome interactions. Unlike previous genetic assessment of Arctic cod sampled from the Chukchi Sea, the high levels of genetic diversity found in Arctic cod assayed in this study, across regions, suggests that the species in the Beaufort and Chukchi seas does not suffer from low levels of genetic variation, at least at neutral genetic markers. The large census size of Arctic cod may allow this species to retain high levels of genetic diversity. In addition, we discovered the presence of hybridization between Arctic and Polar cod (although low in frequency). Hybridization is expected to occur when environmental changes modify species distributions that result in contact between species that were previously separated. In such cases, hybridization may be an evolutionary mechanism that promotes an increase in genetic diversity that may provide species occupying changing environments with locally-adapted genotypes and, therefore, phenotypes. Natural selection can only act on the standing genetic variation present within a population. Therefore, given its higher levels of genetic diversity in combination with a large population size, Arctic cod may be resilient to current and future environmental change, as high genetic diversity is expected to increase opportunities for positive selection to act on genetic variants beneficial in different environments, regardless of the source of that genetic variation.

OCS Study

Mineral commodity profiles: nitrogen

Overview -- Nitrogen (N) is an essential element of life and a part of all animal and plant proteins. As a part of the DNA and RNA molecules, nitrogen is an essential constituent of each individual's genetic blueprint. As an essential element in the chlorophyll molecule, nitrogen is vital to a plant's ability to photosynthesize. Some crop plants, such as alfalfa, peas, peanuts, and soybeans, can convert atmospheric nitrogen into a usable form by a process referred to as 'fixation.' Most of the nitrogen that is available for crop production, however, comes from decomposing animal and plant waste or from commercially produced fertilizers. Commercial fertilizers contain nitrogen in the form of ammonium and/or nitrate or in a form that is quickly converted to the ammonium or nitrate form once the fertilizer is applied to the soil. Ammonia is generally the source of nitrogen in fertilizers. Anhydrous ammonia is commercially produced by reacting nitrogen with hydrogen under high temperatures and pressures. The source of nitrogen is the atmosphere, which is almost 80 percent nitrogen. Hydrogen is derived from a variety of raw materials, which include water, and crude oil, coal, and natural gas hydrocarbons. Nitrogen-based fertilizers are produced from ammonia feedstocks through a variety of chemical processes. Small quantities of nitrates are produced from mineral resources principally in Chile. In 2002, anhydrous ammonia and other nitrogen materials were produced in more than 70 countries. Global ammonia production was 108 million metric tons (Mt) of contained nitrogen. With 28 percent of this total, China was the largest producer of ammonia. Asia contributed 46 percent of total world ammonia production, and countries of the former U.S.S.R. represented 13 percent. North America also produced 13 percent of the total; Western Europe, 9 percent; the Middle East, 7 percent; Central America and South America, 5 percent; Eastern Europe, 3 percent; and Africa and Oceania contributed the remaining 4 percent (International Fertilizer Industry Association, 2003b, p. 1-4). In 2002, world ammonia exports were 13.1 Mt of contained nitrogen. Trinidad and Tobago (22 percent), Russia (18 percent), Ukraine (10 percent), and Indonesia (7 percent) accounted for 57 percent of the world total. The largest importing regions were North America with 36 percent of the total followed by Western Europe with 23 percent and Asia with 22 percent (International Fertilizer Industry Association, 2003b, p. 5L-11). In 2002, world urea production was 51.4 Mt of contained nitrogen, and exports were 12.0 Mt of contained nitrogen. China and India, which were the two largest producing countries, accounted for 48 percent of world production. The United States and Canada produced about 10 percent of the total. Russia and Ukraine together accounted for 28 percent of total urea exports; Central America and South America, 27 percent; and Asia, North America, and Western Europe, 10 percent each. North America accounted for 36 percent of the total urea imports; Western Europe, 23 percent; and Asia, 22 percent (International Fertilizer Industry Association, 2003f, p. 1-15). Ammonia production capacity in North America and Western Europe is projected to decline through 2004, and capacity in other world regions is projected to increase. Fluctuating natural gas prices are mainly responsible for the capacity decline in North America. Ammonia production capacity is continuing to shift to world regions that have abundant sources of natural gas, and away from those where costs (raw material, labor, environmental compliance) are higher.

Open-File Report

Hydrogeology and water quality of the Leetown area, West Virginia

The U.S. Geological Survey’s Leetown Science Center and the co-located U.S. Department of Agriculture’s National Center for Cool and Cold Water Aquaculture both depend on large volumes of cold clean ground water to support research operations at their facilities. Currently, ground-water demands are provided by three springs and two standby production wells used to augment supplies during periods of low spring flow. Future expansion of research operations at the Leetown Science Center is dependent on assessing the availability and quality of water to the facilities and in locating prospective sites for additional wells to augment existing water supplies. The hydrogeology of the Leetown area, West Virginia, is a structurally complex karst aquifer. Although the aquifer is a karst system, it is not typical of most highly cavernous karst systems, but is dominated by broad areas of fractured rock drained by a relatively small number of solution conduits. Characterization of the aquifer by use of fluorometric tracer tests, a common approach in most karst terranes, therefore only partly defines the hydrogeologic setting of the area. In order to fully assess the hydrogeology and water quality in the vicinity of Leetown, a multi-disciplinary approach that included both fractured rock and karst research components was needed. The U.S. Geological Survey developed this multi-disciplinary research effort to include geologic, hydrologic, geophysical, geographic, water-quality, and microbiological investigations in order to fully characterize the hydrogeology and water quality of the Leetown area, West Virginia. Detailed geologic and karst mapping provided the framework on which hydrologic investigations were based. Fracture trace and lineament analysis helped locate potential water-bearing fractures and guided installation of monitoring wells. Monitoring wells were drilled for borehole geophysical surveys, water-quality sampling, water-level measurements, and aquifer tests to characterize the quality of water and the hydraulic properties of the aquifer. Surface geophysical surveys provided a 3-dimensional view of bedrock resistivity in order to assess geologic and lithologic controls on ground-water flow. Borehole geophysical surveys were conducted in monitoring wells to assess the storage and movement of water in subsurface fractures. Numerous single-well, multi-well, and straddle packer aquifer tests and step-drawdown tests were conducted to define the hydraulic properties of the aquifer and to assess the role of bedrock fractures and solution conduits in the flow of ground water. Water samples collected from wells and springs were analyzed to assess the current quality of ground water and provide a baseline for future assessment. Microbiological sampling of wells for indicator bacteria and human and animal DNA provided an analysis of agricultural and suburban development impacts on ground-water quality. Light detection and ranging (LiDAR) data were analyzed to develop digital elevation models (DEMs) for assessing sinkhole distribution, to provide elevation data for development of a ground-water flow model, and to assess the distribution of major fractures and faults in the Leetown area. The flow of ground water in the study area is controlled by lithology and geologic structure. Bedrock, especially low permeability units such as the shale Martinsburg Formation and the Conococheague Limestone, act as barriers to water flowing down gradient and across bedding. This retardation of cross-strike flow is especially pronounced in the Leetown area, where bedding typically dips at steep angles. Highly permeable fault and fracture zones that disrupt the rocks in cross-strike directions provide avenues through which ground water can flow laterally across or through strata of low primary permeability. Significant strike parallel thrust faults and cross-strike faults typically coincide with larger solution conduits and act as drains for the more pervasive network of interconnected diffuse fractures. Results of borehole geophysical surveys indicate that although numerous fractures may intersect a borehole, only one or two of the fractures typically transmit most of the water to a well. The diffuse-flow dominated network of fractures that provides the majority of storage occupies only a small proportion of the total aquifer volume but constitutes the majority of porosity within the aquifer. Solution conduits, while occupying a relatively small volume of the overall aquifer, are especially important because they serve as primary drains for the ground-water flow system. Surface resistivity maps and cross-sectionsshow anomalous areas of low resistivities coincident with the prevailing geologic strike at N. 20º E., with major cross-strike faults, and with major springs in the region. Transmissivity derived from straddle packer tests was highly variable, and ranged over three orders of magnitude (1.8 x 10 -6 to 5.9 x 10 -3 ft 2 /d) in diffuse-flow fractures. A similar large variability in transmissivity was documented by single- and multi-well aquifer tests conducted in conduit-flow dominated portions of the aquifer (2.0 x 10 3 to 1.4 x 10 4 ft 2 /d) in lowland areas immediately adjacent to the Leetown Science Center. A stream-gaging station installed on Hopewell Run near the point where the stream exits the Leetown watershed indicates average daily streamflow for the Hopewell Run of approximately 11.2 ft 3 /s, and ranged from a minimum of 1.80 ft 3 /s on September 28, 2005, to a maximum of 73.0 ft 3 /s on December 11, 2003. Base-flow (ground-water) discharge surveys identified numerous small seeps adjacent to streams in the area. Hydrographs of the stage of Balch Spring show rapid response to individual storms. Strong correlation of the flow of Hopewell Run and Balch Spring indicates the nearby losing stream reach is partly responsible for higher fluctuations in the stage of Balch Spring. A water budget for the study period (2003-2005), based on measured precipitation and hydrograph analyses, is expressed as Precipitation (38.60 in/yr) = Surface Runoff (1.36 in/yr) + Ground-Water Discharge (17.73 in/yr) + Evapotranspiration (24.23 in/yr) – Change in storage (4.72 in/yr). Flow of ground water through the epikarst, a shallow zone of intensely weathered rock and regolith, can be rapid (on the order of days or weeks) as flow is concentrated in solution conduits. Flow within the intermediate and deeper zones is typically much slower. Eight dye-tracer tests conducted in the Leetown area found ground-water flow patterns to be divergent, with velocities ranging from about 12.5 to 610 ft/day and a median velocity of 50 ft/day. Estimates of ground-water age in carbonate rocks in the region are on the order of 15 years in the shallower portions of the aquifer to 50 years or older for deeper portions of the aquifer. Shallow springs can have a significant component of fairly young water (< 5 years in age). Ground-water samples collected from 16 sites (12 wells and 4 springs) in the Leetown area were analyzed for more than 340 constituents. Only turbidity, indicator bacteria, and radon were typically present in concentrations exceeding U.S. Environmental Protection Agency (USEPA) drinking-water or aquatic life standards.

West Virginia

Genetic integrity, population status, and long-term viability of isolated populations of shoal bass in the upper Chattahoochee River basin, Georgia

This report characterizes the status of multiple isolated Shoal Bass ( Micropterus cataractae ) populations in the upper Chattahoochee River basin (UCRB), Georgia. The Shoal Bass, a sport fish endemic to the Apalachicola-Chattahoochee-Flint River (ACF) basin, is a fluvial-specialist species considered vulnerable to local extirpations and extinction due to habitat fragmentation and introgression with non-native congeners. Perhaps one of the most isolated populations of Shoal Bass exists in a 2-km reach of Big Creek, a tributary of the Chattahoochee River located near Roswell, Georgia. Big Creek is partially contained within the Chattahoochee River National Recreation Area, although the Big Creek watershed is riddled with urban land cover. Roswell Mill Dam limits the upstream extent of the Shoal Bass population at Big Creek, and the downstream extent is presumably limited to the confluence of Big Creek and the Chattahoochee River. This reach of the Chattahoochee River is thermally depressed because of coldwater releases from Lake Lanier, and is considered unsuitable for Shoal Bass. Herein, we examine the genetic integrity, population status, and long-term viability of the Shoal Bass population in Big Creek. We also examine two additional Shoal Bass populations that occur in the UCRB, specifically the Chestatee River and the upper Chattahoochee River, both of which are impounded at Lake Lanier. Together, the Shoal Bass inhabiting these three stream systems comprise a distinct genetic stock of Shoal Bass (Taylor 2017), underscoring the importance of conserving these populations towards maintaining the overall diversity and adaptive potential of the species. We assessed genetic diversity and estimated effective population sizes within these three rivers by genotyping fish with 16 microsatellite DNA markers. Results demonstrated that the Shoal Bass population in Big Creek has experienced high rates of introgression with non-native Smallmouth Bass (M. dolomieu), purportedly introduced into the Chattahoochee River in the past 10-15 years. Alarmingly, only 24% (15 of 62) of putative Shoal Bass collected from Big Creek were genetically pure Shoal Bass, whereas the majority of fish were first-filial (F1) generation hybrids and unidirectional backcrosses towards Shoal Bass. Fleeting opportunity may remain to conserve the native genome of the Shoal Bass population in Big Creek. High hybridization rates prevented genetic diversity analysis for the Big Creek population. Shoal Bass populations in the Chestatee and Chattahoochee rivers displayed levels of genetic diversity similar to populations that persist in other rivers in the ACF basin, namely the Flint and Chipola rivers. Effective population sizes of 93.8– 197.4 for the Chestatee and Chattahoochee rivers (combined) suggest that the conservation status of these populations is stable for the short-term, but may be at risk of losing genetic diversity and adaptive potential in the long-term. To estimate age and mortality of the three populations, we used fish scales and capture-markrecapture (CMR) as complementary, non-lethal methods for age estimation. Estimated ages of phenotypic Shoal Bass ranged from 1-12 years in all three populations, demonstrating increased longevity compared to populations elsewhere within the native range. Catch-curve estimates of annual mortality ranged from 18.4-23.7%, which are markedly lower than those observed in other Shoal Bass populations in the ACF basin. These differences in life-history characteristics underscore the need for the development of population-specific management and conservation strategies for Shoal Bass in the UCRB. The lowest recruitment variability (i.e., the variation in year-class strength) was observed in the Chestatee River, a forested watershed, whereas the highest variability was observed in Big Creek, an urbanized watershed. Recruitment strength in Big Creek was negatively influenced by discharge variability in the summer months, suggesting that flashy, sediment-laden flows hinder survival of recently hatched young. Other statistically significant models from Big Creek and the Chattahoochee River indicated that over-winter survival could be an important pinch-point for recruitment in UCRB populations. A multi-agency sampling effort was conducted from May 2013-May 2016 to estimate the population size of Shoal Bass occupying the 1-km of wadeable shoal habitats in Big Creek. Using CMR models, we estimated that approximately 219-348 Shoal Bass (≥ 70 mm total length) occupied the area throughout the duration of our study. These estimates largely reflect abundance of individuals aged 0-2 years, as only 9% (36 of 408) tagged fish were aged ≥ 7 years. Local abundance appeared similar to that reported for a population that inhabited Little Uchee Creek, a similar-sized tributary of the Chattahoochee River, prior to its recent functional extirpation. The low abundance of large, adult Shoal Bass further suggests the long-term viability of the Big Creek population may be in jeopardy. Perhaps most importantly, CMR estimates reflect abundance of phenotypic Shoal Bass – genetic analyses suggest the abundance of pure Shoal Bass could be an order of magnitude smaller. To evaluate the potential for adult Shoal Bass to emigrate from Big Creek into the mainstem Chattahoochee River, we tagged eight adults with acoustic telemetry tags and assessed their seasonal residency at two stationary receiver locations located in increasing proximity to the confluence with the Chattahoochee River. Fish took up residency near the confluence during the fall and winter months, during which time water temperatures in Big Creek were periodically colder than the Chattahoochee River. Although we were unable to document emigration, we conclude that the potential for emigration is highest during the winter months when the Chattahoochee River may be warmer than Big Creek. Two of the tagged fish were caught by anglers near the confluence, suggesting that angling pressure at Big Creek may be higher than previously suspected. Overall, this study observed unique life-history characteristics and characterized the population status of multiple Shoal Bass populations in the UCRB. Populations in the Chestatee and Chattahoochee rivers appear stable at present and likely represent the last remaining strongholds for pure Shoal Bass in the UCRB. Efforts to preserve forested watershed conditions, natural hydrology, and shoal habitats would contribute to the long-term persistence of Shoal Bass populations in these two rivers. Additionally, the detection of non-native Alabama Bass and their associated hybrids in both rivers is cause for concern. Diligent monitoring of hybridization dynamics between Alabama Bass and Shoal Bass is warranted, along with an assessment of Alabama Bass invasion extent upstream of Lake Lanier. The Shoal Bass population in Big Creek is threatened by elevated levels of introgression with nonnative Smallmouth Bass, recruitment variability, low abundance of adults, and isolation from other populations. Conservation intervention is urgently needed to restore and preserve this genetically distinct population, which would contribute to preservation of range wide genetic diversity and adaptability of the species. Additionally, an urban sport fishery for Shoal Bass at Big Creek has the potential to serve as a tool for increasing public awareness, engagement, and support of Shoal Bass conservation efforts in the UCRB. We suggest strategies for conservation of the remnant shoal habitats and Shoal Bass population in Big Creek, including potential development of a supplemental stocking program, selective removal of non-native congeners, and delivery of environmental education programs that could bolster awareness and appreciation.

Georgia

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