Practical evaluation of carbon sources of forest soils in Slovenia from stable and radio-carbon isotope measurements

Carbon sources were estimated by measuring carbon isotope ratios (??¹³C and ??¹⁴C) with accelerator mass spectrometry (AMS) in forest soils of different lithology. Six locations were selected in temperate deciduous and coniferous stands in Slovenia (?irovski vrh, Idrija, Ko?evski Rog, Pohorje, Gori?nica, and Rakitna), where carbonate rocks consisting of limestone and dolomite are abundant as underlying bedrock. Carbon isotope fractionation would not have occurred in two carbonaceous soils, since the values of both ??¹³C and ??¹⁴C changed consistently in these soils after thermal (550°C, 2?h) or chemical (1?M HCl, 24?h) treatments. Organic components were found to be predominant carbon sources (70?C100%) in the uppermost portions (0?C2?cm in depth). In deeper portions at a depth of about 30?C35?cm, soil carbon may be derived completely from underlying carbonate minerals in Idria, western part of Slovenia. The Combination of heat and chemical treatments with AMS provides practical information on soil carbon sources in carbonaceous soils.     

Concentrations and possible sources of PAHs in sediments from Bohai Bay and adjacent shelf

Concentrations, spatial distribution and sources of 16 polycyclic aromatic hydrocarbons (PAHs) listed by the United States Environmental Protection Agency as priority pollutants were investigated in surface sediments of Bohai Bay, North China. Total concentrations of PAHs were in the range of 140.6–300.7 ng/g (dry wt), with an average of 188.0 ng/g. The three predominant PAHs were phenanthrene, acenaphthene and naphthalene. Sedimentary PAH concentrations of the north and central Bohai Bay were higher than those of the southern side of this bay. PAHs source analysis suggested that PAHs in most of the sediments were mainly from grass, wood and coal incomplete combustion. At other stations near the estuaries (Luanhe River Estuary and Chaohe River Estuary) or the oil drilling platform, both petrogenic and pyrogenic inputs were significant. The pyrogenic PAHs close to the oil drilling platform were mainly from petroleum combustion.     

Quantification of organic pollutant degradation in contaminated aquifers using compound specific stable isotope analysis - Review of recent developments

Compound specific stable isotope analysis (CSIA) has been established as a viable tool for proving, characterizing and assessing degradation of organic pollutants within contaminated aquifers. The fractionation of stable isotopes during contaminant degradation leads to observable shifts in stable isotope ratios which can serve as an indicator for in situ pollutant degradation and allow for a quantitative assessment by means of the so-called Rayleigh (distillation) equation.This review highlights the recent developments of the Rayleigh equation approach for quantifying in situ degradation of organic pollutants in contaminated aquifers. The advantages and limitations of the Rayleigh equation approach are discussed and suggestions for improvements are given. Concepts are provided to estimate the uncertainty due to errors or variability of input parameters and how to deal with such uncertainty. Moreover, the applicability of the Rayleigh equation approach is evaluated regarding the heterogeneity and complexity of groundwater systems. For such systems, the review discusses the relevance of non-destructive processes, which affect the concentration (e.g., dispersive mixing) and potentially also the stable isotope ratio of contaminants (e.g., sorption, volatilization), and the resulting implications for the Rayleigh equation approach.     

A theoretical model of the stable sulfur isotope distribution in marine sediments

A diffusion-diagenesis model of the sulfur cycle is developed to calculate theoretical distributions of stable sulfur isotopes in marine sediments. The model describes the depth variation in δ³⁴S of dissolved sulfate and H₂S. and of pyrite. The effects of sulfate reduction, sulfate and H₂S diffusion. and of sedimentation are considered as well as the bacterial isotope fractionation and the degree of pyrite formation. Under open system conditions of sulfur diagenesis the isotopic difference, ΔSO^2?₄ — H₂S, tends to increase with depth being smaller than the bacterial fractionation factor near the sediment surface and larger in deeper layers. The two isotopes in SO^2?₄ or in H₂S do not diffuse in the same proportion as they occur in the porewater. This explains why sulfur, which is incorporated from seawater sulfate by diffusion and precipitation as pyrite, can be enriched in ³²S relative to the seawater sulfate. The model calculations demonstrate the importance of taking the whole dynamic sulfur cycle into account before drawing conclusions about sulfur diagenesis from the stable isotope distribution.     

Compositional differences in enstatite chondrites based on carbon and nitrogen stable isotope measurements

Carbon and nitrogen abundance and isotopic compositions, from four EH4, one EH5, five EL6 chondrites and one aubrite, were determined by using stepped pyrolysis (N only) and combustion (N and C) extractions in attempts to distinguish the components present. Carbon contents range from 0.15 to 0.70 wt%, with no systematic relationship between carbon content and meteorite group or petrologic type. Whole-rock δ¹³C values range from −28.5 to −4.1 %., Most C occurs as graphite and when temperature steps above 700°C are considered, there is a difference between EH4,5 (δ¹³C = −9.1 to -5.8%.) and EL6 chondrites (δ¹³C = −6.7 to +4.2%.). Carbon in Bustee aubrite is isotopically lighter (δ¹³C = −24%.) than in any enstatite chondrite.

Nitrogen occurs as osbornite, sinoite and in isostructural substitution for oxygen in silicate lattice sites. Nitrogen abundances and isotopic compositions are more variable than C, due to the heterogenous distribution of N-bearing minerals. Three EL6's containing osbornite have higher N concentrations than other type 6 enstatite chondrites. Sinoite, where present, is depleted in ¹⁵N relative to osbornite. Nitrogen in the Bustee aubrite has a similar abundance and δ¹⁵N value to those of EL6's, again dominated by the presence of osbornite.

In addition to the refractory C-and N-bearing minerals there is also organic material (largely terrestrial contamination) and evidence for at least two “exotic” components. The first is a host for Xe (HL) and is characterized by δ¹³C <-−47%. and δ¹⁵N ≤−73%., whereas the second is less well-defined, but is marked by δ¹⁵N = +269%.     

Estimation of lead sources in a Japanese cedar ecosystem using stable isotope analysis

Anthropogenic Pb affects the environment worldwide. To understand its effect on forest ecosystem, Pb isotope ratios were determined in precipitation, various components of vegetation, the forest floor, soil and parent material in a Japanese cedar (Cryptomeria japonica D. Don) forest stand. The average ²⁰⁶Pb/²⁰⁷Pb ratio in bulk precipitation was 1.14 ± 0.01 (mean ± SD), whereas that in the subsoil (20–130 cm) was 1.18 ± 0.01. Intermediate ratios ranging from 1.15 to 1.16 were observed in the vegetation, the forest floor, and the surface soil (0–10 cm). Using the ²⁰⁶Pb/²⁰⁷Pb ratios, the contribution of anthropogenic sources to Pb accumulated in the forest were estimated by the simple binary mixing model. Sixty-two percent of the Pb in the forest floor, 71% in the vegetation, and 55% in the surface soil (0–10 cm) originated from anthropogenic sources, but only 16% in the sub-surface soil (10–20 cm) was anthropogenic. These results suggest that internal Pb cycling occurs mainly between surface soil and vegetation in a Japanese cedar ecosystem, and that anthropogenic Pb strongly influences Pb cycling. Although the Japanese cedar ecosystem has a shallow forest floor, very little atmospherically derived Pb migrated downward over 10 cm in depth.     

Tracing diffuse anthropogenic Pb sources in rural soils by means of Pb isotope analysis

Knowledge of the cause and source of Pb pollution is important to abate environmental Pb pollution by taking source-related actions. Lead isotope analysis is a potentially powerful tool to identify anthropogenic Pb and its sources in the environment. Spatial information on the variation of anthropogenic Pb content and anthropogenic Pb sources in rural topsoils is remarkably limited. This study presents results of a survey of approximately 350 topsoil samples from rural locations covering the entire Netherlands, for which the bulk geochemical and Pb isotope compositions were determined. The specific aim of this study is to determine the anthropogenic Pb sources in the topsoils from rural areas in The Netherlands. The spatial distribution of anthropogenic Pb in soils in The Netherlands will be explained in terms of land use and pollution sources.Nearly all studied topsoils display Pb contents that exceed the amount expected based on the soil lithology. The range in Pb isotope ratios of the additional Pb fraction in rural Dutch topsoils is established at 1.056–1.199, 2.336–2.486 and 0.452–0.490 for ²⁰⁶Pb/²⁰⁷Pb, ²⁰⁷Pb/²⁰⁸Pb and ²⁰⁶Pb/²⁰⁸Pb, respectively. Five land use types are distinguished (forest, open nature, moor, arable land and grassland) with distinct isotopic compositions for added Pb. Additional Pb in soils of natural areas (forest, open nature and moor) has on average lower ²⁰⁶Pb/²⁰⁷Pb, ²⁰⁸Pb/²⁰⁷Pb and ²⁰⁶Pb/²⁰⁸Pb ratios than the agricultural soils (arable land and grassland). Additional Pb in both natural area soils and agricultural soils is interpreted to be of anthropogenic origin: most likely a mixture of coal/galena, incinerator ashes and gasoline Pb. The dominant sources of additional Pb in the topsoil of open nature areas are most likely incinerator ash and gasoline Pb. In contrast, the on average higher ²⁰⁶Pb/²⁰⁷Pb, ²⁰⁸Pb/²⁰⁷Pb and ²⁰⁶Pb/²⁰⁸Pb ratios of additional Pb in agricultural soils are most likely caused by the presence of animal manure and N–P fertilizers.Several areas are observed with notably high additional Pb contents (26–211 mg/kg on an organic matter-free basis) in the topsoil. The largest area is the Randstad area, which has the highest population and traffic density, and hosts a considerable fraction of the Dutch chemical industry. Two other areas with high additional Pb contents in the topsoil are located near the Dutch borders and are most likely influenced by German and Belgian chemical industries. The topsoils in the coastal dunes and southern, central and northern forests are characterized by relatively low additional Pb contents (<10 mg/kg on an organic matter-free basis). The population, traffic and chemical industry density is low in these areas and no fertilizers are applied.     

Chemical characteristics and pollution sources of petroleum hydrocarbons and PAHs in sediments from the Beiluohe River,Northern China

To determine the degree of hydrocarbon contamination and the contribution of local petroleum industries to contaminant loadings in sediments from the Beiluohe River, China, 12 surface sediment samples were collected for geochemical analysis in 2005. Sediment samples were extracted by organic solvents, separated by silica gel column chromatography and the profiles of n-alkanes, biomarkers and polycyclic aromatic hydrocarbons (PAHs) in sediments were analyzed by gas chromatography with flame ionization detector and gas chromatography/mass spectroscopy. Concentrations of total hydrocarbons in the sediments varied from 12.1 to 3,761.5 μg g⁻¹ dry wt, indicating that most sediments in Beiluohe River was only slightly to moderately contaminated by hydrocarbons. Concentrations of PAHs for six samples (sum of 16 isomers) varied from 17.7 to 407.7 ng g⁻¹ dry wt and at present low levels of PAHs did not cause adverse biological effects in Beiluohe River sedimentary environment. PAH compositions, n-alkanes and biomarker profiles all suggested that there were different sources of contaminations in studied areas. n-Alkanes reflect two distinct sources: a fossil n-alkane series from crude oil at sites S40, S43, S87 and plantwax n-alkanes at sites S39 and S45. Judged by their PAH ratios, the sediments at site S15 were pyrolytic, sediments at S17 and S43 were petrogenic, and sediments at S39, S40 and S64 had a mixture source of pyrolytic and petrogenic.     

The relationship between total dissolved carbon dioxide and its stable carbon isotope ratio in aquatic sediments

A model is constructed to predict the stable carbon isotope ratio of the total dissolved CO₂ in aquatic sediments and laboratory reactors. The major parameters of the model are the fractionation between CO₂ and CH₄ as well as the intra-molecular fractionation of acetate, the relative production of CH₄ from CO₂ reduction versus acetate fermentation, the net production ratio of CO₂ to CH₄ and the stable carbon isotope ratio of the source organic carbon. The model is fitted to published data and to date from the littoral sediments of Lake Memphremagog, Quebec, Canada. The inclusion of the intra-molecular fractionation factor of acetate in the model provides a good fit to the data; without this factor, the values of the other parameters necessary for a good fit appear unreasonable.     

应用稳定同位素判别治岭头金银矿床的成矿流体性质和来源

浙江治岭头金银矿床中两期含金石英脉的氢、氧、碳及硫的稳定同位素研究结果表明各自具有明显不同的同位素组成特征,两期矿化作用的成矿热液流体具不同成因类型。早期是与基底陈蔡群变质水有关的热液流体;而晚期则是与盖层火山岩浆水和古大气降水有关的混合热液流体。因此治岭头矿床是一个具有多源成矿热液流体相叠加的复合型矿床。     

Stable carbon isotope evidence for two sources of organic matter in coastal sediments: seagrasses and plankton

Organic carbon from sediments collected in Texas seagrass meadows was enriched in ¹³C by an average of 6.6% relative to organic carbon from offshore sediments. Within the South Texas hay system examined. δ¹³C values became increasingly more typical of offshore sediments with increasing distance from seagrass meadows. This permits the use of carbon isotope data as a measure of the relative contributions of seagrasses and plankton to sedimentary organic matter.     

Vertical patterns of stable carbon isotope in soils and particle-size fractions of karst areas, Southwest China

Taking limestone soil and yellow soil, the two major soil types in karst areas as examples, analyzing stable carbon isotope composition (δ¹³C value) of soil organic matter (SOM) in bulk soils and particle-size fractions of four soil profiles under three vegetable forms, the following results are reached: in the limestone soil profile, soil organic carbon contents are all above 1.0%, the highest value is 7.1% in the surface soil; however, they are between 0.3% and 4.6% in the three yellow soil profiles. From the surface to the bottom of the soil profiles, the variation of δ¹³C value of soil organic carbon for limestone soil profile is only between −24.1‰ and −23.0‰, however, it’s between −24.8‰ and −21.1‰ for yellow soil profiles. The variation range of δ¹³C value of soil organic carbon associated with particle-size separates is slight for limestone soil but is considerable for yellow soil. The contrast research indicates that the changes between the contents and the δ¹³C value of soil organic carbon with depth are complex. The vertical patterns of stable carbon isotope in soil organic matter have a distinct regional characteristic in karst areas.     

Degradation of methyl bromide and methyl chloride in soil microcosms: Use of stable C isotope fractionation and stable isotope probing to identify reactions and the responsible microorganisms

Bacteria in soil microcosm experiments oxidized elevated levels of methyl chloride (MeCl) and methyl bromide (MeBr), the former compound more rapidly than the latter. MeBr was also removed by chemical reactions while MeCl was not. Chemical degradation dominated the early removal of MeBr and accounted for more than half of its total loss. Fractionation of stable carbon isotopes during chemical degradation of MeBr resulted in a kinetic isotope effect (KIE) of 59 ± 7‰. Soil bacterial oxidation dominated the later removal of MeBr and MeCl and was characterized by different KIEs for each compound. The KIE for MeBr oxidation was 69 ± 9‰ and the KIE for MeCl oxidation was 49 ± 3‰. Stable isotope probing revealed that different populations of soil bacteria assimilated added ¹³C-labeled MeBr and MeCl. The identity of the active MeBr and MeCl degrading bacteria in soil was determined by analysis of 16S rRNA gene sequences amplified from ¹³C-DNA fractions, which identified a number of sequences from organisms not previously thought to be involved in methyl halide degradation. These included Burkholderia, the major clone type in the ¹³C-MeBr fraction, and Rhodobacter, Lysobacter and Nocardioides the major clone types in the ¹³C-MeCl fraction. None of the 16S rRNA gene sequences for methyl halide oxidizing bacteria currently in culture (including Aminobacter strain IMB-1 isolated from fumigated soil) were identified. Functional gene clone types closely related to Aminobacter spp. were identified in libraries containing the sequences for the cmuA gene, which codes for the enzyme known to catalyze the initial step in the oxidation of MeBr and MeCl. The cmuA gene was limited to members of the alpha-Proteobacteria whereas the greater diversity demonstrated by the 16S rRNA gene may indicate that other enzymes catalyze methyl halide oxidation in different groups of bacteria.     

滇西北红山—红牛铜矿成矿物源:矿物学和稳定同位素约束

本文研究了中甸弧南段红山—红牛铜矿主要夕卡岩矿物石榴石和辉石成因矿物学特征,显示矿区石榴石为钙铁-钙铝石榴石系列,早期多为钙铁榴石,晚期为钙铝榴石。矿区部分石榴石在背散射光下可以观察到明显的环带结构,钙铝榴石和钙铁榴石含量差别越大环带越明显。矿区辉石主要为次透辉石和少量的透辉石,表明夕卡岩早期成矿流体为高温、酸性和高氧逸度环境。矿区黄铁矿强富铁亏硫且Co/Ni1,黄铜矿S、Fe和Cu含量较稳定只含有少量的C、Ni和Se,均指示为高温岩浆热液成因。金属硫化物34S众值出现在3.8‰~5.6‰,同时代的石英二长斑岩34S在4.7‰~7.8‰,表明硫主要来自石英二长斑岩。金属矿物和石英二长斑岩铅同位素特征表明,铅主要来自上地壳。石榴石δ~(18)O_(V-SMOW)=6‰~8.8‰,成矿流体δ~(18) OH_2O=4.6‰~7.8‰,反映成矿流体主要来自岩浆热液,且成矿流体存在明显的沸腾作用。夕卡岩矿体中的方解石和围岩方解石C-O同位素研究,指示块状硫化物中的方解石来自岩浆流体,含有硫化物细脉的围岩大理岩中的方解石来自围岩的再结晶作用。     

The application of C isotope measurements to the identification of the sources of C in diamonds: a review

In the past decade, the isotopic compositions of C in > 600 inclusion-bearing diamonds have been determined. Such analyses have revealed the following isotopic characteristics: (1) peridotitic diamonds, which typically contain garnet, chromite, olivine and/or orthopyroxene inclusions with refractory compositions (high Mg, Cr), have δ¹³C values predominantly between −10 and −1‰, with a sharp peak in the distribution near −5‰; (2) eclogitic diamonds, which commonly contain inclusions of omphacitic clinopyroxene, Cr-poor pyrope, and/or eclogitic accessory minerals such as rutile, kyanite, coesite or sanidine, have δ¹³C values between −34 and +3‰, with a smaller peak near −5‰; (3) the isotopic compositions obtained for suites of diamonds from individual occurrences are, in general, unique and do not resemble the range and distribution obtained by amalgamating the diamond isotope data from a number of localities; (4) isotopic zoning patterns and heterogeneities are found in some diamonds; cores of coated diamonds tend to be depleted in¹³C relative to the rims, and within single octahedral diamonds δ¹³C variations of nearly 6‰ have been reported.Because expected C isotope fractionations at mantle temperatures are small, attempts to model the full range of diamond isotope values through fractionating a homogeneous mantle C source have been unsuccessful. Nevertheless, fractionation is probably responsible for some of the observed variation in δ¹³C values. Two other models have also been proposed to account for the diamond characteristics outlined above. The “primordial model” suggests that the range and distribution of C isotope compositions are inherited from primordial C in the mantle which has an inhomogeneous isotopic composition, such as that found in meteorites. The “subduction model” suggests that subducted, crustal C is the source of C in diamonds, as organic and inorganic C compounds in the crust exhibit a range of δ¹³C values similar to that observed in diamonds. This paper reviews the C isotope characteristics of diamonds and compares the models which have been proposed to explain the origins of these characteristics.     

Levels and sources of polycyclic aromatic hydrocarbons (PAHs) in selected irrigated urban agricultural soils in Accra, Ghana

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous organic pollutants in urban environments including urban soils. Elevated concentrations of PAHs in urban soils are caused by incomplete combustion of petroleum and coal. This study assesses 16 individual PAH compounds in a total of 112 surficial soil samples. The objective was to assess and compare the levels of contamination as well as examine the main sources of PAHs in four urban agricultural soils using molecular ratios of some specific hydrocarbons. The study showed that PAH levels in soil ranged from 1.23 ng/kg in soil collected from Dzorwulu to 2.95 ng/kg in soil collected from Ghana Broadcasting Cooperation (GBC) vegetable irrigation site. Of the total PAHs, the more water soluble PAHs (2–4 rings), which tend to be concentrated in the vapour phase were found to dominate the soils. The percentage dominance were Dzorwulu (52.8 %), Marine Drive (62.5 %), CSIR (53.2 %) and GBC (49.2 %). However, there were significant levels of the more carcinogenic PAHs (5–6 rings) present with percentages as 47.1, 37.5 46.8 and 50.8 % for Dzorwulu, Marine Drive, CSIR and GBC vegetable irrigation sites, respectively, and therefore, may impact negatively on public health. Based on the classification by the Institute of Soil Science and Plant Cultivation in Pulawy, Poland, urban soils in Accra could be classified as contaminated to different levels. Molecular ratios of Flu/pyr and PA/Ant were calculated to determine the main sources of PAHs. Results showed that PAHs could originate mainly from incomplete combustion of petroleum products, especially from atmospheric fallout from automobile exhausts. The study further showed that B(a)P concentration of 0.05 ng/kg in soil from GBC urban vegetable irrigation site requires immediate clean-up exercise and monitoring to mitigate human health impact.     

The significance of isotope specific diffusion coefficients for reaction-transport models of sulfate reduction in marine sediments

Modeling isotopic signatures in systems affected by diffusion, advection, and a reaction which modifies the isotopic abundance of a given species, is a discipline in its infancy. Traditionally, much emphasis has been placed on kinetic isotope effects during biochemical reactions, while isotope effects caused by isotope specific diffusion coefficients have been neglected. A recent study by Donahue et al. (2008) suggested that transport related isotope effects may be of similar magnitude as microbially mediated isotope effects. Although it was later shown that the assumed differences in the isotope specific diffusion coefficients were probably overstated by one or two orders of magnitude (Bourg, 2008), this study raises several important issues: (1) Is it possible to directly calculate isotopic enrichment factors from measured concentration data without modeling the respective system? (2) Do changes in porosity and advection velocity modulate the influence of isotope specific diffusion coefficients on the fractionation factor α? (3) If one has no a priori knowledge whether diffusion coefficients are isotope specific or not, what is the nature and magnitude of the error introduced by either assumption? Here we argue (A) That the direct substitution of measured data into a differential equation is problematic and cannot be used as a replacement for a reaction-transport model; (B) That the transport related fractionation scales linearly with the difference between the respective diffusion coefficients of a given isotope system, but depends in a complex non-linear way on the interplay between advection velocity, and downcore changes of temperature and porosity. Last but not least, we argue that the influence of isotope specific diffusion coefficients on microbially mediated sulfate reduction in typical marine sediments is considerably smaller than the error associated with the determination of the fractionation factor.     

Validation of a rapid and simple method for the preparation of aqueous organic compounds prior to compound specific isotope analysis

Landfill leachate comprises a complex mixture of contaminants many of which may have multiple sources in the environment confounding conventional techniques for apportioning sources. Compound specific isotope ratio mass spectrometry offers the potential to “fingerprint” compounds enabling discrimination to be made between different sources. This paper presents a rapid and highly reproducible method to prepare landfill leachate samples for compound specific isotope analysis. A suitable solid phase extraction (SPE) method was developed using artificial contaminant solutions, natural waters spiked with artificial contaminants (eucalyptol, dodecane, benzothiazole, dibutyl phthalate and naphthalene), and samples of landfill leachate. The elution of adsorbed compounds from the SPE cartridge was found to be the major cause of hydrogen and carbon isotope fractionation. Three different SPE cartridge types were tested: C18, Strata-X and ENV+. Fractionation of between 1‰ and 245‰ for hydrogen isotopes and 0–1.83‰ for carbon isotopes was observed. Part of the fractionation could be attributed to the different SPE cartridges but the major contribution was from the choice of the eluting solvent. Our results indicated hydroxylated styrene-divinylbenzene cartridges eluted with dichloromethane resulted in negligible hydrogen or carbon isotope fractionation for any of the tested organic compounds.The isotopic fractionation of hydrogen and/or carbon of most artificial contaminants were proportional to the efficiency with which they were extracted from water. Only naphthalene exhibited isotopic fractionation unrelated to its extraction efficiency but this fractionation was negligible.The comparative extraction of landfill leachate using SPE and LLE indicated SPE generally extracts slightly more of each compound from leachate than LLE. This relationship is stronger than for polar compounds. In addition, the δD composition of each compound prepared using SPE was with two standard deviations of the sample compound prepared using LLE, and the δ¹³C composition of each compound prepare using either method was mostly within two standard deviations.     

Quantification of carbon flow from stable isotope fractionation in rice field soils with different organic matter content

Rice fields are an important source for the greenhouse gas methane produced by acetoclastic and hydrogenotrophic methanogenesis. Fractionation of ¹³C/¹²C can in principle be used to quantify the relative contribution of these pathways, but our knowledge of isotopic fractionation during reduction of CO₂ and turnover of acetate in different methanogenic environments is still scarce. We therefore measured δ¹³C signatures in two types of anoxic Italian rice field soils, one with high and one with low degradable organic matter (OM) content. Both soils were incubated in the presence and absence of methyl fluoride, a specific inhibitor of acetoclastic methanogenesis. Optimization of methyl fluoride concentration resulted in complete inhibition of acetoclastic methanogenesis. CH₄ was then exclusively produced by hydrogenotrophic methanogenesis, allowing determination of the isotopic signatures and fractionation factors specific for this methanogenic pathway. Acetate, which was then no longer consumed, accumulated and was used for determination of the isotopic signature of the fermentatively produced acetate (both total acetate and methyl carbon of acetate). Hence, all isotopic signatures, including fractionation factors were determined for the methanogenic soil. These data, were then used for computation of the relative contribution of the two methanogenic pathways. In the high OM soil, the contribution of acetoclastic methanogenesis to total CH₄ production increased simultaneously with decreasing acetate concentration. In the low OM soil, methanogenesis from H₂/CO₂ was clearly greater than theoretically expected. Furthermore, isotope fractionation of hydrogenotrophic methanogenesis indicated that the in situ energy status of methanogens strongly depended on the availability of organic carbon in the rice field soil system. Collectively, our data show that the study of isotopic fractionation in methanogenic environments allows a deeper insight into the ongoing processes, which may be quite different in the same ecosystem with different content of degradable OM.