Evaluation of petroleum hydrocarbon biodegradation in shallow groundwater by hydrogeochemical indicators and C, S-isotopes

Biodegradation is one of the main natural attenuation processes in groundwater contaminated with petroleum hydrocarbons. In this work, preliminary studies have been carried out by analyzing the concentrations of total petroleum hydrocarbons (TPH), dissolved inorganic carbon (DIC), dominant terminal electron accepters or donors, as well as δ ¹³C_DIC and δ ³⁴S_SO4, to reveal the biodegradation mechanism of petroleum hydrocarbons in a contaminated site. The results show that along groundwater flow in the central line of the plume, the concentrations of electron acceptors, pH, and E _h increased but TPH and DIC decreased. The δ ¹³C_DIC values of the contaminated groundwater were in the range of ?14.02 to ?22.28 ‰_PDB and ?7.71 to 8.36 ‰_PDB, which reflected a significant depletion and enrichment of ¹³C, respectively. The increase of DIC is believed to result from the non-methanogenic and methanogenic biodegradation of petroleum hydrocarbon in groundwater. Meanwhile, from the contaminated source to the downgradient of the plume, the ³⁴S in the contaminated groundwater became more depleted. The Rayleigh model calculation confirmed the occurrence of bacterial sulfate reduction as a biodegradation pathway of the petroleum hydrocarbon in the contaminated aquifers. It was concluded that stable isotope measurements, combined with other biogeochemical measurements, can be a useful tool to prove the occurrence of the biodegradation process and to identify the dominant terminal electron-accepting process in contaminated aquifers.     

Enhancement of in situ biodegradation of organic compounds in groundwater by targeted pump and treat intervention

This study demonstrates the value of targeted pump and treatment (PAT) to enhance the in situ biodegradation of organic contaminants in groundwater for improved restoration. The approach is illustrated for a plume of phenolic compounds in a sandstone aquifer, where PAT is used for hydraulic containment and removal of dissolved phase contaminants from specific depth intervals. Time-series analysis of the plume hydrochemistry and stable isotope composition of dissolved species (δ³⁴S-SO₄, δ¹³C-CH₄, δ¹³C-TDIC (TDIC = Total Dissolved Inorganic Carbon)) in groundwater samples from high-resolution multilevel samplers were used to deduce changes in the relative significance of biodegradation processes and microbial activity in the plume, induced by the PAT system over 3 years. The PAT system has reduced the maximum contaminant concentrations (up to 6800 mg L⁻¹ total phenols) in the plume by 50% to ∼70% at different locations. This intervention has (i) stimulated in situ biodegradation in general, with an approximate doubling of contaminant turnover based on TDIC concentration, which has increased from <200 mg L⁻¹ to >350 mg L⁻¹, (ii) enhanced the activity of SO₄-reducing microorganisms (marked by a declining SO₄ concentration with corresponding increase in SO₄-δ³⁴S to values >7–14‰_V-CDT relative to background values of 1.9–6.5‰_V-CDT), and (iii) where the TDIC increase is greatest, has changed TDIC-δ¹³C from values of −10 to −15‰_V-PDB to ∼−20‰_V-PDB. This indicates an increase in the relative importance of respiration processes (including denitrification and anaerobic methane oxidation, AMO) that yield ¹³C-depleted TDIC over fermentation and acetoclastic methanogenesis that yield ¹³C-enriched TDIC in the plume, leading to higher contaminant turnover. The plume fringe was found to be a zone of enhanced biodegradation by SO₄-reduction and methanogenesis. Isotopically heavy methane compositions (up to −47.8‰_V-PDB) and trends between δ¹³C-TDIC and δ¹³C-CH₄ suggest that AMO occurs at the plume fringe where the contaminant concentrations have been reduced by the PAT system. Mass and isotope balances for inorganic carbon in the plume confirm the shift in spatial dominance of different biodegradation processes and significant increase in contribution of anaerobic respiration for contaminant biodegradation in zones targeted by the PAT system. The enhanced in situ biodegradation results from a reduction in organic contaminant concentrations in the plume to levels below those that formerly suppressed microbial activity, combined with increased supply of soluble electron acceptors (e.g. nitrate) into the plume by dispersion. An interruption of the PAT system and recovery of the dissolved organic contaminant concentrations towards former values highlights the dynamic nature of this enhancement on restoration and relatively rapid response of the aquifer microorganisms to changing conditions induced by the PAT system. In situ restoration using this combined engineered and passive approach has the potential to manage plumes of biodegradable contaminants over shorter timescales than would be possible using these methods independently. The application of PAT in this way strongly depends on the ability to ensure an adequate flux of dissolved electron acceptors into the plume by advection and dispersion, particularly in heterogeneous aquifers.     

Concentration trends and water-level fluctuations at underground storage tank sites

Concentration trends of monitor wells utilized in monitored natural attenuation at petroleum underground storage tank sites can be used to predict achievement of regulatory standards if the data approximate a first-order decline trend. However, declining concentration trends often display seasonal and other fluctuations that complicate trend interpretation. Seasonal correlations between concentration and water-level elevation, including in-phase and inverse relationships, constitute one of the most common types of variation. The in-phase fluctuations are most common for monitor wells located in or near the source area of the release. This relationship may be the result of increased contact with the smear zone in the source area during periods of high water table. Conversely, inverse trends of water-level elevation and concentration are most common in downgradient wells beyond the limit of the source area. In a year long study of short-term fluctuations in BTEX and other parameters in a downgradient monitor well, the data suggest that the winter/spring recharge event significantly controls the concentration trends of BTEX as well as inorganic compounds in the well. Recharge and associated water table rise began in late fall and were soon followed by a slug of inorganic ions strongly influenced by road salt application. This slug of recharge diluted the concentrations of petroleum compounds and alkalinity (bicarbonate). Electron acceptors including oxygen, nitrate, and sulfate, which is a component of road salt, are also contributed to the water table during recharge. Oxygen and nitrate were not detected in the monitor well samples and were most likely consumed quickly in biodegradation reactions at the top of the contaminant plume. Sulfate peaked during winter/spring recharge and then slowly declined during the summer and fall, along with redox potential. Alkalinity (bicarbonate) increased during this period, which may represent the coupled oxidation of organic carbon to CO₂ with sulfate as the electron acceptor. BTEX concentrations peaked in the fall probably due to the lack of diluting recharge. The slow changes in concentration over the summer and fall months, interpreted to be caused by biodegradation, contrast with the rapid changes associated with dilution during the recharge event.     

地下水苯系物微生物降解及其碳同位素标记

微生物降解是地下水中有机物自然衰减评估的关键,单体稳定同位素是一种有效的评估方法。在对某油罐泄露场地地下水流场识别的基础上,刻画不同地下水中污染物、微生物及电子受体特征,发现随着与污染源水力联系的减弱,污染物浓度明显减小,微生物群落结构和电子受体氧化还原作用类型与源相似的程度也逐渐减弱,呈现出“污染源-下游源区-下游污染羽-上游源区-侧翼污染羽”的空间变化规律。甲苯、间/对二甲苯碳的同位素标记结果发现,降解程度“侧翼污染羽﹥下游污染羽﹥下游源区”,与电子受体表征降解量的排序相反;该场地微生物降解符合一般化学反应“勒沙特列原理”:污染物浓度越高,降解量越大,但降解程度相对减小。     

Diel behavior of stable isotopes of dissolved oxygen and dissolved inorganic carbon in rivers over a range of trophic conditions,and in a mesocosm experiment

Rates of diel (24-h) biogeochemical processes in rivers and their effect on daily changes in the concentration of metals and metalloids have been well documented in the literature over the last 20 years. Investigations into the effects of these processes on aquatic systems and the underlying mechanisms that control the processes can significantly improve our understanding of how natural aquatic environments function and will respond to changing environmental conditions and anthropogenic impacts. Daily changes in the rates of biogeochemical processes have, more recently, been shown to influence the stable isotope composition of dissolved oxygen and dissolved inorganic carbon in natural waters. Here we present a comprehensive picture of the persistence and reproducibility of diel cycles of the ¹⁸O composition of dissolved molecular oxygen (δ¹⁸O-DO) and the ¹³C composition of dissolved inorganic carbon (δ¹³C-DIC) across five Montana, USA rivers investigated over a 4-year period. A mesocosm experiment showed the same behavior in δ¹⁸O-DO and δ¹³C-DIC as seen in riverine settings across light and dark periods.A cross plot of δ¹⁸O-DO and δ¹³C-DIC from each stream exhibits a clockwise elliptical pattern which is attributed to the daily changes in the balance of metabolic rates as well as air–water gas exchange. The amplitude of the change in the isotope composition is shown to be directly related to the trophic state of the river and a relationship between net productivity and diel changes in δ¹⁸O-DO and δ¹³C-DIC is presented. This relationship between trophic status with δ¹⁸O-DO, δ¹³C-DIC and production emphasizes the significance of how rates of biogeochemical processes in natural systems can influence the daily changes in the composition of surface waters.     

Evaluating the sulfur isotopic composition of biodegraded petroleum: The case of the Western Canada Sedimentary Basin

The sulfur isotopic composition (δ³⁴S) of petroleum is believed to be affected mainly by sulfur incorporation reactions into the sedimentary organic matter during the early diagenesis. However, secondary processes could affect the original δ³⁴S of oil under the effect of thermal maturity or of the microbial activity of biodegraded reservoirs. In this study, the different processes that may affect the δ³⁴S of in-reservoir oils were assessed based on the sulfur content and isotopes of a series of oil and core samples coming from various reservoirs of the Lower Cretaceous Mannville Group, Western Canada Sedimentary Basin (WCSB). Based on the molecular study, these samples appear to have reached various levels of maturity and biodegradation, ranging from 0 to 6.5 on the biodegradation scale of Peters and Moldowan. In addition, mixing of organic matter coming from different source rocks was identified based on the comparison with extensive correlation studies performed in the WCSB.Investigation of the δ³⁴S shows a trend that seems a priori correlated to the level of biodegradation. However, a careful interpretation of molecular and sulfur isotope data leads to the conclusion that the observed δ³⁴S variations have rather to be ascribed to contributions of oils generated by various source rocks. Alternatively, variations of δ³⁴S could neither be related to maturity differences nor to kinetic effects during organic sulfur compounds biodegradation. In the case of some specific core samples showing a common origin based on biomarker study, δ³⁴S variations might not be related to different sources but to secondary sulfur incorporation/exchange processes occurring within the reservoir. These processes would involve reduced sulfur species from bacterial sulfate reduction formed in situ or migrated into Mannville reservoirs. This hypothesis is supported by laboratory experiments showing sulfur exchange/incorporation under plausible conditions for shallow reservoirs.     

The effect of aquifer heterogeneity on natural attenuation rate of BTEX

Monitored natural attenuation can be a viable option for remediation of groundwater contamination by BTEX compounds. Under the field conditions, the rate of contaminant mass attenuation through natural processes, such as biodegradation, to a large extent affected by the groundwater flow regime, which is primarily controlled by the aquifer heterogeneity. Numerical simulation techniques were used to describe quantitatively the relationship between biodegradation rate of BTEX and aquifer heterogeneity. Different levels of aquifer heterogeneity were described by random hydraulic conductivity fields (K) having different statistical parameters, the coefficient of variation (CV) and the correlation length (h). The Turning Bands Algorithm was used to generate such K fields. Visual MODFLOW/RT3D was used to simulate the fate and transport of dissolved BTEX plume within heterogeneous aquifers. The multispecies reactive transport approach described BTEX degradation using multiple terminal electron-accepting processes. First-order biodegradation rate constants were calculated from simulated BTEX plumes in heterogeneous flow fields. The results showed that aquifer heterogeneity significantly affected biodegradation rate; it decreased with increasing CV when h was in the range of up to 12 m, whereas it increased with increasing CV when h was greater than about 12 m. For well characterized aquifers, this finding could be of great value in assessing the effectiveness of natural attenuation during feasibility studies at BTEX contaminated sites.     

Field investigation of the natural attenuation and intrinsic biodegradation rates at an underground storage tank site

 Contamination of groundwater by petroleum-hydrocarbons is a widespread environmental problem. Natural attenuation is a passive remedial approach to degrade and dissipate contaminants in soil and groundwater. In this study, a mass flux approach was used to calculate the contaminant mass reduction and field-scale decay rate at a gasoline spill site. The mass flux technique is accomplished using the differences in total contaminant mass flux across two cross sections of the contaminant plume. The mass flux calculation shows that up to 88% of the dissolved BTEX (benzene, toluene, ethylbenzene, and xylene isomers) removal was observed by natural attenuation processes. The efficiency of intrinsic biodegradation was evaluated by the in situ tracer method. A first-order decay model was applied for the natural attenuation and intrinsic biodegradation rate calculation. Results reveal that intrinsic biodegradation process was the major cause of the BTEX reduction among the natural attenuation mechanisms, and iron reduction was the dominant biodegradation pattern within the plume. Approximately 87% of the BTEX removal was caused by intrinsic biodegradation processes. The calculated BTEX natural attenuation and intrinsic biodegradation rates were 0.24 and 0.16% l/day, respectively. Results suggest that natural attenuation mechanisms can effectively contain the plume, and the mass flux method is useful in assessing the efficiency of the natural attenuation. Received: 6 December 1999 · Accepted: 11 July 2000     

Assessment of MTBE biodegradation in contaminated groundwater using C and C analysis: Field and laboratory microcosm studies

Radiolabelled assays and compound-specific stable isotope analysis (CSIA) were used to assess methyl tert-butyl ether (MTBE) biodegradation in an unleaded fuel plume in a UK chalk aquifer, both in the field and in laboratory microcosm experiments. The ¹⁴C-MTBE radiorespirometry studies demonstrated widespread potential for aerobic and anaerobic MTBE biodegradation in the aquifer. However, δ¹³C compositions of MTBE in groundwater samples from the plume showed no significant ¹³C enrichment that would indicate MTBE biodegradation at the field scale. Carbon isotope enrichment during MTBE biodegradation was assessed in the microcosms when dissolved O₂ was not limiting, compared with low in situ concentrations (2 mg/L) in the aquifer, and in the absence of O₂. The microcosm experiments showed ubiquitous potential for aerobic MTBE biodegradation in the aquifer within hundreds of days. Aerobic MTBE biodegradation in the microcosms produced an enrichment of 7‰ in the MTBE δ¹³C composition and an isotope enrichment factor (ε) of −1.53‰ when dissolved O₂ was not limiting. However, for the low dissolved O₂ concentration of up to 2 mg/L that characterizes most of the MTBE plume fringe, aerobic MTBE biodegradation produced an enrichment of 0.5-0.7‰, corresponding to an ε value of −0.22‰ to −0.24‰. No anaerobic MTBE biodegradation occurred under these experimental conditions. These results suggest the existence of a complex MTBE-biodegrading community in the aquifer, which may consist of different aerobic species competing for MTBE and dissolved O₂. Under low O₂ conditions, the lower fractionating species have been shown to govern overall MTBE C-isotope fractionation during biodegradation, confirming the results of previous laboratory experiments mixing pure cultures. This implies that significant aerobic MTBE biodegradation could occur under the low dissolved O₂ concentration that typifies the reactive fringe zone of MTBE plumes, without producing detectable changes in the MTBE δ¹³C composition. This observed insensitivity of C isotope enrichment to MTBE biodegradation could lead to significant underestimation of aerobic MTBE biodegradation at field scale, with an unnecessarily pessimistic performance assessment for natural attenuation. Site-specific C isotope enrichment factors are, therefore, required to reliably quantify MTBE biodegradation, which may limit CSIA as a tool for the in situ assessment of MTBE biodegradation in groundwater using only C isotopes.     

Rates of microbial hydrogen oxidation and sulfate reduction in Opalinus Clay rock

Hydrogen gas (H₂) may be produced by the anoxic corrosion of steel components in underground structures, such as geological repositories for radioactive waste. In such environments, hydrogen was shown to serve as an electron donor for autotrophic bacteria. High gas overpressures are to be avoided in radioactive waste repositories and, thus, microbial consumption of H₂ is generally viewed as beneficial. However, to fully consider this biological process in models of repository evolution over time, it is crucial to determine the in situ rates of microbial hydrogen oxidation and sulfate reduction. These rates were estimated through two distinct in situ experiments, using several measurement and calculation methods. Volumetric consumption rates were calculated to be between 1.13 and 1.93 μmol cm⁻³ day⁻¹ for H₂, and 0.14 and 0.20 μmol cm⁻³ day⁻¹ for sulfate. Based on the stoichiometry of the reaction, there is an excess of H₂ consumed, suggesting that it serves as an electron donor to reduce electron acceptors other than sulfate, and/or that some H₂ is lost via diffusion. These rate estimates are critical to evaluate whether biological H₂ consumption can negate H₂ production in repositories, and to determine whether sulfate reduction can consume sulfate faster than it is replenished by diffusion, which could lead to methanogenic conditions.     

Intrinsic bioremediation of MTBE-contaminated groundwater at a petroleum-hydrocarbon spill site

An oil-refining plant site located in southern Taiwan has been identified as a petroleum-hydrocarbon [mainly methyl tert-butyl ether (MTBE) and benzene, toluene, ethylbenzene, and xylenes (BTEX)] spill site. In this study, groundwater samples collected from the site were analyzed to assess the occurrence of intrinsic MTBE biodegradation. Microcosm experiments were conducted to evaluate the feasibility of biodegrading MTBE by indigenous microorganisms under aerobic, cometabolic, iron reducing, and methanogenic conditions. Results from the field investigation and microbial enumeration indicate that the intrinsic biodegradation of MTBE and BTEX is occurring and causing the decrease in MTBE and BTEX concentrations. Microcosm results show that the indigenous microorganisms were able to biodegrade MTBE under aerobic conditions using MTBE as the sole primary substrate. The detected biodegradation byproduct, tri-butyl alcohol (TBA), can also be biodegraded by the indigenous microorganisms. In addition, microcosms with site groundwater as the medium solution show higher MTBE biodegradation rate. This indicates that the site groundwater might contain some trace minerals or organics, which could enhance the MTBE biodegradation. Results show that the addition of BTEX at low levels could also enhance the MTBE removal. No MTBE removal was detected in iron reducing and methanogenic microcosms. This might be due to the effects of low dissolved oxygen (approximately 0.3 mg/L) within the plume. The low iron reducers and methanogens (<1.8×10³ cell/g of soil) observed in the aquifer also indicate that the iron reduction and methanogenesis are not the dominant biodegradation patterns in the contaminant plume. Results from the microcosm study reveal that preliminary laboratory study is required to determine the appropriate substrates and oxidation-reduction conditions to enhance the biodegradation of MTBE. Results suggest that in situ or on-site aerobic bioremediation using indigenous microorganisms would be a feasible technology to clean up this MTBE-contaminated site.     

Correlation of 13C12C and 34S32S secular variations

Statistical evaluation of 3056 δ¹³C measurements in carbonate rocks and fossils shows that they record a 2‰ ¹³C depletion from the late Proterozoic to the early Paleozoic, a 2.5‰ enrichment to the Permian, and a 1.5‰ depletion to the Cenozoic. These variations, not controlled primarily by facies or alteration phenomena, correlate negatively with the δ³⁴S sulfate secular trend, as confirmed by collation of 1083 δ³⁴S measurements. The correlation suggests that the biologically mediated redox fluxes of the C and S cycles have been approximately balanced through this long span of geological time, generally levelling available oxygen. Such a redox system is consistent with the controlling mechanism proposed by Garrels and Perry (1974). Consequently, the sedimentary reservoirs of C_organic as well as S_{bacteriological'}have varied through geological time.     

Delta Chromium-53/52 isotopic composition of native and contaminated groundwater,Mojave Desert,USA

Chromium(VI) concentrations in groundwater sampled from three contaminant plumes in aquifers in the Mojave Desert near Hinkley, Topock and El Mirage, California, USA, were as high as 2600, 5800 and 330 μg/L, respectively. δ^53/52Cr compositions from more than 50 samples collected within these plumes ranged from near 0‰ to almost 4‰ near the plume margins. Assuming only reductive fractionation of Cr(VI) to Cr(III) within the plume, apparent fractionation factors for δ^53/52Cr isotopes ranged from ε_app = 0.3 to 0.4 within the Hinkley and Topock plumes, respectively, and only the El Mirage plume had a fractionation factor similar to the laboratory derived value of ε = 3.5. One possible explanation for the difference between field and laboratory fractionation factors at the Hinkley and Topock sites is localized reductive fractionation of Cr(VI) to Cr(III), with subsequent advective mixing of native and contaminated water near the plume margin. Chromium(VI) concentrations and δ^53/52Cr isotopic compositions did not uniquely define the source of Cr near the plume margin, or the extent of reductive fractionation within the plume. However, Cr(VI) and δ^53/52Cr data contribute to understanding of the interaction between reductive and mixing processes that occur within and near the margins of Cr contamination plumes. Reductive fractionation of Cr(VI) predominates in plumes having higher ε_app, these plumes may be suitable for monitored natural attenuation. In contrast, advective mixing predominates in plumes having lower ε_app, the highly dispersed margins of these plumes may be difficult to define and manage.     

Geochemical heterogeneity and isotope geochemistry of natural attenuation processes in a gasoline-contaminated aquifer at the Hnevice site,Czech Republic

The geochemical processes, water–rock interactions and stable isotopes distribution (δ¹³C of DIC and δ¹⁸O and δ³⁴S of \({\text{SO}}^{{{\text{2 - }}}}_{{\text{4}}} \)) were investigated in the gasoline-contaminated aquifer at the Hnevice site, 50  km northwest of Prague, Czech Republic. Diesel, gasoline and oil leaks originate from a large fuel storage area causing heavy contamination of the saturated and unsaturated zones in an area of about 0.7  km². Groundwater investigations were conducted using five multilevel sampler wells with emphasis on redox parameters and degradation by-products and a solid-phase study focused on iron speciation and determination of principal and secondary minerals. Based on the study of groundwater and solid-phase geochemistry, four different geochemical zones were described. Zone I is thought to be background consisting of an aerobic aquifer and the absence of reduced species in significant concentrations. Zone II is situated in the plume core with methanogenic, sulphate and iron-reducing conditions accompanied by ankerite and kutnahorite precipitates and significant depletion of the oxidation capacity of the aquifer. Zone III is a mixing (corona) zone, situated at the fringe of the plume with high biodegradation rates and Fe(III)-precipitants. In zone IV, reoxidation of Fe(II) minerals (with e.g. the occurrence of psilomelane and cornelite) is typical.     

石灰岩含水介质中苯系物（BTEX） 生物可降解性判识

    岩溶含水系统遭受石油烃污染的环境问题十分普遍。相对于多孔含水介质,石油烃BTEX在石灰岩含水介质中的生 物可降解性还不确定。为此,本研究开展了BTEX在石灰石和岩溶地下水介质中的静态微元体实验。经过77天的实验检测 分析,结果表明：（1） BTEX化合物在可利用电子受体溶解氧或硝酸盐存在条件下具有生物可降解性;（2） 向系统中补充 电子受体硝酸盐,具有促进生物降解的作用,其对BTEX的去除率可高达94%;（3） 未发现补充硫酸盐能够促进BTEX生物 可降解性;（4） 甲苯和二甲苯容易被生物降解,但苯的去除具有一定的难度。     

Limitations in the use of compound-specific stable isotope analysis to understand the behaviour of a complex BTEX groundwater contamination near Brussels (Belgium)

The application of compound-specific stable isotope analysis (CSIA) was evaluated to characterise a complex groundwater contamination. For this purpose, δ¹³C and δ²H analysis of benzenes and alkylated derivatives were used to interpret both the impact of different sources on a contaminant plume and the presence of degradation processes. The different contaminant sources could be distinguished based on their combined δ¹³C–δ²H signature of the benzene, toluene, ethylbenzene and xylenes (BTEX) dissolved in the groundwater. Despite this source differentiation, plume characterisation was not possible due to the complex mixing of the respective contaminant plumes. Furthermore, the original isotope signatures of the sources were not preserved across these plumes. To estimate the level of in situ biodegradation independently from concentration data, the Rayleigh equation was used. Although current literature identifies the application of CSIA as very promising in the frame of characterising organic groundwater pollution, this study has indicated that this approach can be limited with respect to successfully distinguish the different plumes and their relation to the known source zones.     

Calcium isotopes in a proglacial weathering environment: Damma glacier, Switzerland

The biogeochemical cycling and isotopic fractionation of calcium during the initial stages of weathering were investigated in an alpine soil chronosequence (Damma glacier, Switzerland). This site has a homogeneous silicate lithology and minimal biological impacts due to sparse vegetation cover. Calcium isotopic compositions, obtained by TIMS using a ⁴³Ca-⁴⁶Ca double spike, were measured in the main Ca pools. During this very early stage of weathering, the young soils which have formed (δ^44/42Ca=+0.44‰) were indistinguishable to the rocks from which they were derived (δ^44/42Ca=+0.44‰) and stream water (δ^44/42Ca=+0.48‰) was also within error of the average rock. This lack of variation indicates that the dissolution of the bulk silicate rock does not strongly fractionate Ca isotopes. The only Ca pool which was strongly fractionated from bulk rock was vegetation, which exhibited an enrichment of light Ca isotopes. Significant Ca isotope fractionation between bulk rock and the dissolved flux of Ca is likely to only occur where the Ca biogeochemical cycle is dominated by secondary processes such as biological cycling, adsorption and secondary mineral precipitation.     

Influencing factors and significance of organic and inorganic nitrogen isotopic compositions in lacustrine sedimentary rocks

Comprehensive nitrogen biogeochemical cycle has been reconstructed for representative lacustrine organic-rich sedimentary rock in China, namely the Triassic Yanchang Formation (YF, 199–230 Ma) in Ordos and the Cretaceous Qingshankou Formation (QF, 86–92 Ma) in Songliao basins, by evaluating the organic and inorganic nitrogen isotopic compositions rather than only organic or bulk nitrogen isotopic compositions. The results indicate that the nitrogen isotope values of bulk rock (δ¹⁵N_bulk) in the non-metamorphic stage are significantly different from that of kerogen, which challenge the conceptual framework of sedimentary nitrogen isotope interpretation. The δ¹⁵N_bulk from the YF and QF were lower than their respective the nitrogen isotope values of kerogen (δ¹⁵N_ker), with offsets up to ～5.1‰, which have the inverse relationship for the metamorphosed rock. Thermal evolution did not significantly modify the δ¹⁵N of bulk rock and kerogen. The δ¹⁵N of sediments from the YF (δ¹⁵N_bulk, 1.6‰–5.6‰) were lower than that of rock from the QF (δ¹⁵N_bulk, 10.2‰–15.3‰). The nitrogen isotope values of silicate incorporated nitrogen (δ¹⁵N_sil) were slightly lower than those of the δ¹⁵N_ker in the YF and obviously lower for the QF. The fact that different nitrogen cycles occur in the YF and QF due to the different depositional redox conditions leads to different isotopic results. The YF water environment dominated by oxic conditions is not conducive to the occurrence of denitrification and anammox, and no abundant N₂ loss leads to the relatively light δ¹⁵N_bulk. In the stratified water for the QF, redox transition zone promotes denitrification and anammox, resulting in the heavy δ¹⁵N_bulk of rock and promotes the DNRA, resulting in heavy δ¹⁵N_ker and low δ¹⁵N_sil.     

Effect of contaminant concentration on in situ bacterial sulfate reduction and methanogenesis in phenol-contaminated groundwater

The availability of dissolved O₂ can limit biodegradation of organic compounds in aquifers. Where O₂ is depleted, biodegradation proceeds via anaerobic processes, including NO₃-, Mn(IV)-, Fe(III)- and SO₄-reduction and fermentation/methanogenesis. The environmental controls on these anaerobic processes must be understood to support implementation of management strategies such as monitored natural attenuation (MNA). In this study stable isotope analysis is used to show that the relative significance of two key anaerobic biodegradation processes (bacterial SO₄ reduction (BSR) and methanogenesis) in a phenol-contaminated sandstone aquifer is sensitive to spatial and temporal changes in total dissolved phenols concentration (TPC) (= phenol + cresols + dimethylphenols) over a 5-a period. In general, ³⁴SO₄-enrichment (characteristic of bacterial SO₄ reduction) is restricted spatially to locations where TPC < 2000 mg L⁻¹. In contrast, ¹³C-depleted CH₄ and ¹³C-enriched CO₂ isotope compositions (characteristic of methanogenesis) were measured at TPC up to 8000 mg L⁻¹. This is consistent with previous studies that demonstrate suppression of BSR at TPC of >500 mg L⁻¹, and suggests that methanogenic microorganisms may have a higher tolerance for TPC in this contaminant plume. It is concluded that isotopic enrichment trends can be used to identify conditions under which in situ biodegradation may be limited by the properties of the biodegradation substrate (in this case TPC). Such data may be used to deduce the performance of MNA for contaminated groundwater in similar settings.     

Thermal analysis (TG–DTA) and isotopic characterization (13C–15N) of humic acids from different origins

Thermal analyses (TG–DTA), elemental composition and isotope analyses (¹³C and ¹⁵N) were performed on humic acids (HA) from peats, leonardites and lignites, in order to investigate their structure and the changes taking place during the humification process. Thermal analyses showed structural differences between HA samples in relation to their coalification rank. In particular the lignite HA were characterized by a more stable chemical composition at high temperatures.The δ¹³C and δ¹⁵N values can provide information on the biogeochemical processes involved in HA formation. In particular, peat HA were linked to anoxic environments that enable plant residues to persist in their structure. In contrast, leonardite and lignite HA formation seems to be governed by different biogeochemical processes from those responsible for peat diagenesis. However, the isotopic analyses did not provide any distinction between leonardite and lignite HA. On the basis of the data presented in this study, it may be concluded that TG–DTA and isotope ratio measurements are powerful tools for investigating the formation pathway of humic substances from coals.