Tracking the sources of nitrate in groundwater using coupled nitrogen and boron isotopes: a synthesis

Nitrate (NO3) is one of the world's major pollutants of drinking water resources. Although recent European Directives have reduced input from intensive agriculture, NO3 levels in groundwater are approaching the drinking water limit of 50 mg L(-1) almost everywhere. Determining the sources of groundwater contamination is an important first step toward improving its quality by emission control. It is with this aim that we review here the benefit of using a coupled isotopic approach (delta15N and delta11B), in addition to conventional hydrogeological analyses, to trace the origin of NO3 in water. The studied watersheds include both fractured bedrock and alluvial (subsurface and deep) hydrogeological contexts. The joint use of nitrogen and boron isotope systematics in each context deciphers the origin of NO3 in the groundwater and allows a semi-quantification of the contributions of the respective pollution sources (mineral fertilizers, wastewater, and animal manure).     

Identifying the sources of subsurface contamination at the Hanford Site in Washington using high-precision uranium isotopic measurements

In the mid-1990s, a groundwater plume of uranium (U) was detected in monitoring wells in the B-BX-BY Waste Management Area at the Hanford Site in Washington. This area has been used since the late 1940s to store high-level radioactive waste and other products of U fuel-rod processing. Using multiple-collector ICP source magnetic sector mass spectrometry, high-precision uranium isotopic analyses were conducted of samples of vadose zone contamination and of groundwater. The isotope ratios 236U/238U, 234U/238U, and 238U/235U are used to distinguish contaminant sources. On the basis of the isotopic data, the source of the groundwater contamination appears to be related to a 1951 overflow event at tank BX-102 that spilled high-level U waste into the vadose zone. The U isotopic variation of the groundwater plume is a result of mixing between contaminant U from this spill and natural background U. Vadose zone U contamination at tank B-110 likely predates the recorded tank leak and can be ruled out as a significant source of groundwater contamination, based on the U isotopic composition. The locus of vadose zone contamination is displaced from the initial locus of groundwater contamination, indicating that lateral migration in the vadose zone was at least 8 times greater than vertical migration. The time evolution of the groundwater plume suggests an average U migration rate of approximately 0.7-0.8 m/day showing slight retardation relative to a groundwater flow of approximately 1 m/day.     

Spectroscopic and diffraction study of uranium speciation in contaminated vadose zone sediments from the Hanford site, Washington state

Contamination of vadose zone sediments under tank BX-102 at the Hanford site, Washington, resulted from the accidental release of 7-8 metric tons of uranium dissolved in caustic aqueous sludge in 1951. We have applied synchrotron-based X-ray spectroscopic and diffraction techniques to characterize the speciation of uranium in samples of these contaminated sediments. UIII-edge X-ray absorption fine structure (XAFS) spectroscopic studies demonstrate that uranium occurs predominantly as a uranium(VI) silicate from the uranophane group of minerals. XAFS cannot distinguish between the members of this mineral group due to the near identical local coordination environments of uranium in these phases. However, these phases differ crystallographically, and can be distinguished using X-ray diffraction (XRD) methods. As the concentration of uranium was too low for conventional XRD to detect these phases, X-ray microdiffraction (microXRD) was used to collect diffraction patterns on approximately 20 microm diameter areas of localized high uranium concentration found using microscanning X-ray fluorescence (microSXRF). Only sodium boltwoodite, Na(UO2)(SiO3OH) x 1.5H20, was observed; no other uranophane group minerals were present. Sodium boltwoodite formation has effectively sequestered uranium in these sediments under the current geochemical and hydrologic conditions. Attempts to remediate the uranium contamination will likely face significant difficulties because of the speciation and distribution of uranium in the sediments.     

Distribution and retention of 137Cs in sediments at the Hanford Site, Washington

137Cesium and other contaminants have leaked from single-shell storage tanks (SSTs) into coarse-textured, relatively unweathered unconsolidated sediments. Contaminated sediments were retrieved from beneath a leaky SST to investigate the distribution of adsorbed 137Cs+ across different sediment size fractions. All fractions contained mica (biotite, muscovite, vermiculatized biotite), quartz, and plagioclase along with smectite and kaolinite in the clay-size fraction. A phosphor-plate autoradiograph method was used to identify particular sediment particles responsible for retaining 137Cs+. The Cs-bearing particles were found to be individual mica flakes or agglomerated smectite, mica, quartz, and plagioclase. Of these, only the micaceous component was capable of sorbing Cs+ strongly. Sorbed 137Cs+ could not be significantly removed from sediments by leaching with dithionite citrate buffer or KOH, but a fraction of the sorbed 137Cs+ (5-22%) was desorbable with solutions containing an excess of Rb+. The small amount of 137Cs+ that might be mobilized by migrating fluids in the future would likely sorb to nearby micaceous clasts in downgradient sediments.     

Evaluating regional patterns in nitrate sources to watersheds in National Parks of the Rocky Mountains using nitrate isotopes

In the Rocky Mountains, there is uncertainty about the source areas and emission types that contribute to nitrate (NO3) deposition, which can adversely affect sensitive aquatic habitats of high-elevation watersheds. Regional patterns in NO3 deposition sources were evaluated using NO3 isotopes in five National Parks, including 37 lakes and 7 precipitation sites. Results indicate that lake NO3 ranged from detection limit to 38 microeq/L, delta18O (NO3) ranged from -5.7 to +21.3% per thousand, and delta15N (NO3) ranged from -6.6 to +4.6 per thousand. delta18O (NO3) in precipitation ranged from +71 to +78% per thousand. delta15N (NO3) in precipitation and lakes overlap; however, delta15N (NO3) in precipitation is more depleted than delta15N (NO3) in lakes, ranging from -5.5 to -2.0 per thousand. delta15N (NO3) values are significantly related (p < 0.05) to wet deposition of inorganic N, sulfate, and acidity, suggesting that spatial variability of delta15N (NO3) over the Rocky Mountains may be related to source areas of these solutes. Regional patterns show that NO3 and delta15N (NO3) are more enriched in lakes and precipitation from the southern Rockies and at higher elevations compared to the northern Rockies. The correspondence of high NO3 and enriched delta15N (NO3) in precipitation with high NO3 and enriched delta15N (NO3) in lakes, suggests that deposition of inorganic N in wetfall may affect the amount of NO3 in lakes through a combination of direct and indirect processes such as enhanced nitrification.     

Impacts of land use change on nitrogen cycling archived in semiarid unsaturated zone nitrate profiles, southern High Plains, Texas

Nitrate (NO3) profiles in semiarid unsaturated zones archive land use change (LUC) impacts on nitrogen (N) cycling with implications for agricultural N management and groundwater quality. This study quantified LUC impacts on NO3 inventories and fluxes by measuring NO3 profiles beneath natural and rainfed (nonirrigated) agricultural ecosystems in the southern High Plains (SHP). Inventories of NO3-N under natural ecosystems in the SHP normalized by profile depth are extremely low (2-10 kg NO3-N/ha/m), in contrast to those in many semiarid regions in the southwestern U.S. Many profiles beneath cropland (9 of 19 profiles) have inventories at depth that range from 28-580 kg NO3--N/ha/m (median 135 kg/ha/m) that correspond to initial cultivation, dated using soil water Cl. These inventories represent 74% (median) of the total inventories in these profiles. This NO3 most likely originated from cultivation causing mineralization and nitrification of soil organic nitrogen (SON) in old soil water (precultivation) and is attributed to enhanced microbial activity caused by increased soil wetness beneath cropland (median matric potential -42 m) relative to that beneath natural ecosystems (median -211 m). The SON source is supported by isotopes of NO3 (delta15N: +5.3 to +11.6; delta18O: +3.6 to +12.1). Limited data in South Australia suggest similar processes beneath cropland. Mobilization of the total inventories in these profiles caused by increased drainage/ recharge related to cultivation in the SHP could increase current NO3-N levels in the underlying Ogallala aquifer by an additional 2-26 mg/L (median 17 mg/L).     

Monitoring oxidation of chlorinated ethenes by permanganate in groundwater using stable isotopes: laboratory and field studies

Permanganate injection is increasingly applied for in situ destruction of chlorinated ethenes in groundwater. This laboratory and field study demonstrates the roles that carbon isotope analysis can play in the assessment of oxidation of trichloroethene (TCE) by permanganate. In laboratory experiments a strong carbon isotope fractionation was observed during oxidation of TCE with similar isotopic enrichment factors (-25.1 to -26.8 per thousand) for initial KMnO4 concentrations between 67 and 1,250 mg/L. At the field site, a single permanganate injection episode was conducted in a sandy aquifer contaminated with TCE as dense nonaqueous liquid (DNAPL). After injection, enriched delta13C values of up to +204% and elevated Cl- concentrations were observed at distances of up to 4 m from the injection point. Farther away, the Cl- increased without any change in delta13C of TCE suggesting that Cl- was not produced locally but migrated to the sampling point Except for the closest sampling location to the injection point, the delta13C rebounded to the initial 613C again, likely due to dissolution of DNAPL Isotope mass balance calculations made it possible to identify zones where TCE oxidation continued to occur during the rebound phase. The study indicates that delta13C values can be used to assess the dynamics between TCE oxidation and dissolution and to locate zones of oxidation of chlorinated ethenes that cannot be identified from the Cl- distribution alone.     

Origin assessment of domestic and imported beef sold in the Korean markets using stable carbon and oxygen isotopes

In this study, stable carbon and oxygen isotope data were used to determine the country of origin of imported beef circulating in the Korean market. Using carbon isotopes closely related to the animal diets, Korean beef could easily be distinguished from United States (US) and New Zealand beef. However, oxygen isotopes were more effective in distinguishing the Korean from Australian beef. The oxygen isotopes have specific characteristics reflecting the latitude where cattle feed. Statistical analyses indicate that analyzing both carbon and oxygen isotopes is very useful to assess the country of origin of beef sold in the Korean market. Additionally, we compared the carbon isotopic compositions of defatted and non-defatted beef to evaluate the effect of fat content on carbon isotope values. Our results suggest that by using muscles or tissues that have small amounts of fat, carbon isotope analysis of beef samples may be performed without the inconvenient conventional pretreatment procedure, allowing for easier and faster isotopic analysis.     

Chlorine and carbon isotopes fractionation during volatilization and diffusive transport of trichloroethene in the unsaturated zone

To apply compound-specific isotope methods to the evaluation of the origin and fate of organic contaminants in the unsaturated subsurface, the effect of physicochemical processes on isotope ratios needs to be known. The main objective of this study is to quantify chlorine and carbon isotope fractionation during NAPL-vapor equilibration, air-water partitioning, and diffusion of trichloroethene (TCE) and combinations of these effects during vaporization in porous media. Isotope fractionation is larger during NAPL-vapor equilibration than air-water partitioning. During NAPL-vapor equilibration, carbon, and chlorine isotope ratios evolve in opposite directions although both elements are present in the same bond, with a normal isotope effect for chlorine (ε(Cl) = -0.39 ± 0.03‰) and an inverse effect for carbon (ε(C) = +0.75 ± 0.04‰). During diffusion-controlled vaporization in a sand column, no significant carbon isotope fractionation is observed (ε(C) = +0.10 ± 0.05‰), whereas fairly strong chlorine isotope fractionation occurs (ε(Cl) = -1.39 ± 0.06‰) considering the molecular weight of TCE. In case of carbon, the inverse isotope fractionation associated with NAPL-vapor equilibration and normal diffusion isotope fractionation cancel, whereas for chlorine both processes are accompanied by normal isotope fractionation and hence they cumulate. A source of contamination that aged might thus show a shift toward heavier chlorine isotope ratios.     

西藏青稞碳、氢、氧、氮同位素在不同生态域中差异性研究

目的研究青稞中稳定同位素在不同生态域的差异性。方法 2016年在西藏不同生态域种植不同基因型的青稞品种,利用同位素质谱仪测定不同生态域的青稞样品的稳定碳、氢、氧、氮同位素,结合Kolmogorov-Smirnov和Levene统计量分别检验所有数据的正态性和方差同质性,满足方差齐性时采用LSD多重比较,不满足方差齐性时采用Games-Howell多重比较法进行分析,解析不同生态域青稞样品中稳定碳、氢、氧、氮同位素。结果不同区域种植的青稞δ~(13)C值、δ~(15)N值、δ~(18)O值和δD值差异均极其显著(P0.01)。结论稳定碳、氢、氧、氮同位素指纹可用于青稞的产地溯源,可以进一步研究青稞中多元素组成特征及其用于青稞产地溯源的可行性。     

Tracing of industrial aerosol sources in an urban environment using Pb, Sr, and Nd isotopes

A comprehensive Pb-Sr-Nd isotope tracer study of atmospheric trace metal pollution has been performed in the urban environment of Strasbourg-Kehl. Filter dust of the principal pollutant sources (waste incinerators, thermal power plant and steel plant) and soot of car and ship exhausts have been analyzed. In addition tree barks (as biomonitors) and PM10 have been analyzed to trace and determine the distribution of the pollution in the environment. The industrial sources have highly variable epsilonNd values (-9.7 and -12.5 for incinerators and -17.5 for steel plant). Much higher epsilonNd values have been found for soot of car exhausts (-6 and -6.9). These high values make the Nd isotope system a powerful tool for the discrimination of traffic emissions but especially for the identification of diesel derived particles in the urban environment. The 206Pb/207Pb isotope ratios of gasoline are low (1.089) compared to diesel soot (1.159). The 26Pb/207Pb ratios of 1.151-1.152 for the steel plant and 1.152 for the solid waste incinerator are close to the Pb isotope ratio of diesel. The 87Sr/ 8Sr isotope ratios of the principal industrial sources vary significantly: 0.7095 for the domestic solid waste incinerator, 0.709 for the steel plant, and 0.7087 for car exhaust soot. PM10 aerosols collected in the urban center of Strasbourg show the influence of the pollutant sources at 3-7 km distance from the center. Most of the aerosols Pb isotopic compositions suggest Pb admixtures from at least three sources: a natural background and in function of the wind direction the domestic waste incinerator (S-wind) or the steel plant and the chemical waste incinerator (NE-wind). The traffic contribution can only be estimated with help of Nd isotopes. Therefore the clear identification of different pollutant sources in the urban environment is only possible by combining the three different isotope systems and is based on the fact that significant differences exist between the Pb, Sr, and Nd isotope ratios of the natural atmospheric background and pollutants containing Pb, Sr, and Nd of industrial origin with similar variable 206Pb/207Pb, 87Sr/ 86Sr, and 143Nd/144Nd.     

Sources and migration of plutonium in groundwater at the Savannah River site

The isotopic composition, size distribution, and redox speciation of plutonium (Pu) in the groundwater in the vicinity of the F-area seepage basins at the U.S. Department of Energy Savannah River Site (SRS) were examined. A low 240Pu/239Pu ratio in the upstream control well signifies a Pu source otherthan global fallout and indicates reactor-produced Pu. Elevated 240Pu/239Pu atom ratios downstream from the seepage basins are due to the decay of transplutonium isotopes, mainly 244Cm to 240Pu, which were generated at the SRS. Evidence suggests that the migration of basin-released Pu isotopes is minor. Rather, it is the transplutonium isotopes that migrate preferentially downstream and in the process decay to yield progeny Pu isotopes. Size fractionation studies with cross-flow ultrafiltration show that <4% of the 239Pu or 240Pu is found in the colloidal fraction, a finding that is consistent with the higher Pu oxidation states observed in the SRS groundwater. The observation of a low abundance of colloid-associated Pu in SRS groundwater cannot be extrapolated to all sites, but is in contrast to the conclusions of prior groundwater Pu studies at the SRS and elsewhere. This work is unique in its application of a novel combination of sampling and processing protocols as well as its use of thermal ionization mass spectrometry for the detection of Pu isotopes. This allows quantification of the Pu source terms and better determination of the ambient Pu size and redox speciation representative of in situ conditions.     

Applications, considerations, and sources of uncertainty when using stable isotope analysis in ecotoxicology

Stable isotope analysis (SIA) has become a powerful tool for ecotoxicologists to study dietary exposure and biomagnification of contaminants in wild animal populations. The use of SIA in ecotoxicology continues to expand and, while much more is known about the mechanisms driving patterns of isotopic ratios in consumers, there remain several considerations or sources of uncertainty that can influence interpretation of data from field studies. We outline current uses of SIA in ecotoxicology, including estimating the importance of dietary sources of carbon and their application in biomagnification studies, and we present six main considerations or sources of uncertainty associated with the approach: (1) unequal diet-tissue stable isotope fractionation among species, (2) variable diet-tissue stable isotope fractionation within a given species, (3) different stable isotope ratios in different tissues of the animal, (4) fluctuating baseline stable isotope ratios across systems, (5) the presence of true omnivores, and (6) movement of animals and nutrients between food webs. Since these considerations or sources of uncertainty are difficult to assess in field studies, we advocate that researchers consider the following in designing ecotoxicological research and interpreting results: assess and utilize variation in stable isotope diet-tissue fractionation among animal groups available in the literature; determine stable isotope ratios in multiple tissues to provide a temporal assessment of feeding; adequately characterize baseline isotope ratios; utilize stomach contents when possible; and assess and integrate life history of study animals in a system.     

新、旧世界国家葡萄酒氧稳定同位素时空分布特征研究进展

氧稳定同位素是葡萄酒真实性检测的重要指标之一,其比值在自然因素(葡萄品种、地理环境、气候条件)和人为因素(栽培管理、酿酒工艺)影响下,在不同产地显示出不同时空分布特征。欧盟等旧世界国家已率先建立葡萄酒水中氧稳定同位素的测定方法及数据库,并将其应用于葡萄酒真实性检测中。然而,国内外在葡萄酒氧稳定同位素时空分布特征方面的研究较少,尚未形成统一数据平台,难以实现数据共享。本文总结了新世界(中国、美国、澳大利亚、智利等)和旧世界(西班牙、法国、意大利、奥地利等)国家葡萄酒中水的氧稳定同位素数据分布及鉴别应用,概述了影响该比值的因素,并阐述了目前存在问题和今后发展方向,目的在于明确氧稳定同位素比值在世界主要酿酒国家的分布特征,为我国建立葡萄酒氧同位素数据库及真伪鉴别提供数据支持。     

Pyrogenic inputs of anthropogenic Pb and Hg to sediments of the Hood Canal, Washington, in the 20th century: source evidence from stable Pb isotopes and PAH signatures

Combustion-derived PAHs and stable Pb isotopic signatures ((206)Pb/(207)Pb) in sedimentary records assisted in reconstructing the sources of atmospheric inputs of anthropogenic Pb and Hg to the Hood Canal, Washington. The sediment-focusing corrected peak fluxes of total Pb and Hg (1960-70s) demonstrate that the watershed of Hood Canal has received greater atmospheric inputs of these metals than its mostly rural land use would predict. The tight relationships between the Pb, Hg, and organic markers in the cores indicate that these metals are derived from industrial combustion emissions. Multiple lines of evidence point to the Asarco smelter, located in the Main Basin of Puget Sound, as the major emission source of these metals to the watershed of the Hood Canal. The evidence includes (1) similar PAH isomer ratios in sediment cores from the two basins, (2) the correlations between Pb, Hg, and Cu in sediments and previously studied environmental samples including particulate matter emitted from the Asarco smelter's main stack at the peak of production, and (3) Pb isotope ratios. The natural rate of recovery in Hood Canal since the 1970s, back to preindustrial metal concentrations, was linear and contrasts with recovery rates reported for the Main Basin which slowed post late 1980s.     

稳定同位素结合粗脂肪、粗蛋白含量判别核桃的产地来源

为了判别核桃的产地来源,利用稳定同位素质谱仪测定来自河南、新疆、河北、甘肃、陕西、四川、湖北、云南、广西等省的344个核桃样品的C、H、O、N同位素比值,用元素分析仪测定N元素含量并转化为粗蛋白含量,用理化分析方法测定粗脂肪含量。结果表明,核桃中的δ~2H、δ~(18)O值随纬度的降低而减小;河南省亚地区核桃的δ~(13)C值随海拔的增加而增加。利用线性判别发现,C、H、O、N同位素比值对不同省份、河南省亚地区核桃产地的正确判别率为71. 1%和57. 9%;结合粗脂肪、粗蛋白含量后,正确判别率总体分别提升至79. 6%和68. 6%。说明稳定同位素结合有机成分能提高核桃产地的判别效果,且不同省份间的判别率明显高于亚地区的判别率。     

Characterizing dissolved Cu and Cd uptake in terms of the biotic ligand and biodynamics using enriched stable isotopes

The biotic ligand model considers the biological and geochemical complexities that affect metal exposure. It relates toxicity to the fraction of physiological active sites impacted by reactive metal species. The biodynamic model is a complementary construct that predicts bioaccumulation and assumes that toxicity occurs when influx rates exceed rates of loss and detoxification. In this paper we presume that metal influx rates are mechanistically the resulting processes that characterize transmembrane transport. We use enriched stable isotopes to characterize, both in terms of the biotic ligand and biodynamics, dissolved metal uptake by a freshwater snail at water hardness varying up to 180-fold. Upon 24 h exposure, metal uptake was linear over a range encompassing most environmental concentrations; although saturation kinetics were observed at higher concentrations. Cadmium influx rates correlate with changes in the affinity of the biotic ligand, whereas those of Cu correlate with changes in both site affinity and capacity. A relationship between metal influx rate and ligand character asks whether toxicity is the result of accumulation at the biotic ligand or the rate at which metal is transported by that ligand.     

Influence of different sugar sources,nitrogen sources and inocula on the quality characteristics of apple tea wine

An infusion of tea leaves was prepared using 4 g tea/100 mL of the apple juice, ameliorated with different sugar sources, supplemented with different nitrogen sources and inoculated with 5% incola (Saccharomyces cerevisiae var. ellipsoideus , consortia 1 and consortia 2). The highest rate of fermentation and ethanol were found in the apple tea wine having sucrose as a sugar source, while apple juice concentrate as a sugar source gave the highest total phenols, caffeine content and overall acceptability. Higher alcohols, volatile acidity and antioxidant activity were the highest in honey as a sugar source. Apple tea wine having DAHP as a nitrogen source documented the highest ethanol, caffeine content and overall acceptability, lowest higher alcohols, whereas peptone as a nitrogen source had the highest total phenols. Among the different types of inocula, for most of the parameters there were non‐significant differences. However, must inoculated with S. cerevisiae var. ellipsoideus for the preparation of apple tea wine resulted in the highest fermentation efficiency, ethanol and overall acceptability but the lowest volatile acidity. Apple tea wine prepared by ameliorating the apple tea must with apple juice concentrate, DAHP and inoculated with S. cerevisiae var. ellipsoideus was rated as the best. Copyright © 2017 The Institute of Brewing & Distilling     

Hydroxylated metabolites of beta- and delta-hexachlorocyclohexane: bacterial formation, stereochemical configuration, and occurrence in groundwater at a former production site

Although the use of hexachlorocyclohexane (HCH), one of the most popular insecticides after the Second World War, has been discontinued in many countries, problems remain from former production and waste sites. Despite the widespread occurrence of HCHs, the environmental fate of these compounds is not fully understood. In particular, environmental metabolites of the more persistent beta-HCH and delta-HCH have not been fully identified. Such knowledge, however, is important to follow degradation and environmental fate of the HCHs. In the present study, several hydroxy metabolites that formed during incubation of beta- and delta-HCH with the common soil microorganism Sphingobium indicum B90A were isolated, characterized, and stereochemically identified by gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (NMR). The metabolites were identified as isomeric pentachlorocyclohexanols (B1, D1) and tetrachlorocyclohexane-1,4-diols (B2, D2); delta-HCH additionally formed a tetrachloro-2-cyclohexen-1-ol (D3) and a trichloro-2-cyclohexene-1,4-diol (D4), most likely by hydroxylation of delta-pentachlorocyclohexene (delta-PCCH), initially formed by dehydrochlorination. The dehydrochlorinase LinA was responsible for conversion of delta-HCH into delta-PCCH, and the haloalkane dehalogenase LinB was responsible for the transformation of beta-HCH and delta-HCH into B1 and D1, respectively, and subsequently into B2 and D2, respectively. LinB was also responsible for transforming delta-PCCH into D3 and subsequently into D4. These hydroxylations proceeded in accordance with SN2 type reactions with initial substitution of equatorial Cls and formation of axially hydroxylated stereoisomers. The apparently high reactivity of equatorial Cls in beta- and delta-HCH toward initial hydroxylation by LinB of Sphingobium indicum B90A is remarkable when considering the otherwise usually higher reactivity of axial Cls. Several of these metabolites were detected in groundwater from a former HCH production site in Switzerland. Their presence indicates that these reactions proceed under natural environmental conditions and that the metabolites are of environmental relevance.     

Assessment of indigenous reductive dechlorinating potential at a TCE-contaminated site using microcosms, polymerase chain reaction analysis, and site data

A combination of microcosm studies, polymerase chain reaction (PCR) analysis, and site data was used to assess the indigenous reductive dechlorinating potential in a trichloroethene (TCE)-contaminated aquifer at Cape Canaveral Air Station, Florida. Sediment and groundwater were obtained from two distinct locations approximately 10 m apart. Microcosm studies were performed to assess dechlorinating activity under a variety of nutrient and electron donor amendment conditions. Most live microcosms constructed using material from the first location, near well 9 (W09), were negative for dechlorination. All live microcosms constructed using material from the second location (W06) exhibited dechlorination of TCE to vinyl chloride (VC) and ethene (ETH). DNA encoding 16S ribosomal RNA (rDNA) with a sequence nearly identical with that from Dehalococcoides ethenogenes strain 195 was detected in the active microcosms and in the sediment from W06 with polymerase chain reaction (PCR) using primers targeted to unique regions of Dehalococcoides 16S rDNA. Dehalococcoides was not detected in the autoclaved microcosms from W06, nor in sediment and most microcosms from W09. The results of the microcosm studies and PCR analysis were supported by field data, which indicated significant accumulation of cis-1,2-dichloroethene (cisDCE) and VC at W06, but not at W09. The different microcosm results obtained for the two locations and the spatial variation of positive PCR results indicates heterogeneous distribution of dechlorinating activity and a specific dechlorinating organism, Dehalococcoides, at the site. As both Dehalococcoides and dechlorination activity were similarly, heterogeneously distributed, this suggests that molecular-probing (which could and should be extended in the future to include virtually all known dechlorinators and/or dehalogenases) can provide a relatively quick and facile method for investigating spatial distributions of dechlorinators on-site.