Stable isotope fractionation of toluene under dynamic phase exchange was studied aiming at ascertaining the effects of gas-liquid partitioning and biodegradation of toluene stable isotope composition in liquid-air phase exchange reactors (Laper). The liquid phase consisted of a mixture of aqueous minimal media, a known amount of a mixture of deuterated (toluene-d) and non-deuterated toluene (toluene-h), and bacteria of toluene degrading strain Pseudomonas putida KT2442. During biodegradation experiments, the liquid and air-phase concentrations of both toluene isotopologues were monitored to determine the observable stable isotope fractionation in each phase. The results show a strong fractionation in both phases with apparent enrichment factors beyond −800‰. An offset was observed between enrichment factors in the liquid and the gas phase with gas-phase values showing a stronger fractionation in the gas than in the liquid phase. Numerical simulation and parameter fitting routine was used to challenge hypotheses to explain the unexpected experimental data. The numerical results showed that either a very strong, yet unlikely, fractionation of the phase exchange process or a – so far unreported – direct consumption of gas phase compounds by aqueous phase microorganisms could explain the observed fractionation effects. The observed effect can be of relevance for the analysis of volatile contaminant biodegradation using stable isotope analysis in unsaturated subsurface compartments or other environmental compartment containing a gas and a liquid phase.

Laboratory experiments and point observations, for instance in wetlands, have shown evidence that microbial sulfate reduction (MSR) can lower sulfate and toxic metal concentrations in acid mine drainage (AMD). We here hypothesize that MSR can impact the fate of AMD in entire catchments. To test this, we developed a sulfur isotope fractionation and mass-balance method, and applied it at multiple locations in the catchment of an abandoned copper mine (Nautanen, northern Sweden). Results showed that MSR caused considerable, catchment-scale immobilization of sulfur corresponding to a retention of 27 ± 15% under unfrozen conditions in the summer season, with local values ranging between 13 ± 10% and 53 ± 18%. Present evidence of extensive MSR in Nautanen, together with previous evidence of local MSR occurring under many different conditions, suggest that field-scale MSR is most likely important also at other AMD sites, where retention of AMD may be enhanced through nature-based solutions. More generally, the developed isotope fractionation analysis scheme provides a relatively simple tool for quantification of spatio-temporal trends in MSR, answering to the emerging need of pollution control from cumulative anthropogenic pressures in the landscape, where strategies taking advantage of MSR can provide viable options.

Ammonia (NH₃) volatilized from soils plays an important role in N cycle and air pollution, thus it is important to trace the emission source and predict source contributions to development strategies mitigating the environmental harmful of soil NH₃ volatilization. The measurements of ¹⁵N natural abundance (δ¹⁵N) could be used as a complementary tool for apportioning emissions sources to resolve the contribution of multiple NH₃ emission sources to air NH₃ pollution. However, information of the changes of δ¹⁵N–NH₃ values during the whole volatilization process under different N application rates are currently lacking. Hence, to fill this gap, we conducted a 15-day incubation experiment included different urea-N application rates to determine δ¹⁵N values of NH₃ during volatilization process. Results showed that volatilization process depleted ¹⁵N in NH₃. The average δ¹⁵N value of NH₃ volatilized from the 0, 20, 180, and 360 kg N ha⁻¹ treatment was −16.2 ± 7.3‰, −26.0 ± 5.4‰, −34.8 ± 4.8‰, and −40.6 ± 5.7‰. Overall, δ¹⁵N–NH₃ values ranged from −46.0‰ to −4.7‰ during the whole volatilization process, with lower in higher urea-N application treatments than those in control. δ¹⁵N–NH₃ values during the NH₃ volatilization process were much lower than those of the primary sources, soil (−3.4 ± 0.1‰) and urea (−3.6 ± 0.1‰). Therefore, large isotopic fractionation may occur during soil volatilization process. Moreover, negative relationships between soil NH₄⁺-N and NH₃ volatilization rate and δ¹⁵N–NH₃ values were observed in this study. Our results could be used as evidences of NH₃ source apportionments and N cycle.

Initial Cadmium (Cd) isotope fractionation studies in cereals ascribed the retention of Cd and its light isotopes to the binding of Cd to sulfur (S). To better understand the relation of Cd binding to S and Cd isotope fractionation in soils and plants, we combined isotope and XAS speciation analyses in soil-rice systems that were rich in Cd and S. The systems included distinct water management (flooded vs. non-flooded) and rice accessions with (excluder) and without (non-excluder) functional membrane transporter OsHMA3 that transports Cd into root vacuoles. Initially, 13% of Cd in the soil was bound to S. Through soil flooding, the proportion of Cd bound to S increased to 100%. Soil flooding enriched the rice plants towards heavy isotopes (δ¹¹⁴/¹¹⁰Cd = −0.37 to −0.39%) compared to the plants that grew on non-flooded soils (δ¹¹⁴/¹¹⁰Cd = −0.45 to −0.56%) suggesting that preferentially light Cd isotopes precipitated into Cd sulfides. Isotope compositions in CaCl₂ root extracts indicated that the root surface contributed to the isotope shift between soil and plant during soil flooding. In rice roots, Cd was fully bound to S in all treatments. The roots in the excluder rice strongly retained Cd and its lights isotopes while heavy isotopes were transported to the shoots (Δ¹¹⁴/¹¹⁰Cdₛₕₒₒₜ₋ᵣₒₒₜ 0.16–0.19‰). The non-excluder rice accumulated Cd in shoots and the apparent difference in isotope composition between roots and shoots was smaller than that of the excluder rice (Δ¹¹⁴/¹¹⁰Cdₛₕₒₒₜ₋ᵣₒₒₜ −0.02 to 0.08‰). We ascribe the retention of light Cd isotopes in the roots of the excluder rice to the membrane transport of Cd by OsHMA3 and/or chelating Cd–S complexes in the vacuole. Cd–S was the major binding form in flooded soils and rice roots and partly contributed to the immobilization of Cd and its light isotopes in soil-rice systems.

The determination of both stable nitrogen (δ¹⁵N–NO₃⁻) and stable oxygen (δ¹⁸O–NO₃⁻) isotopic signatures of nitrate in PM₂.₅ has shown potential for an approach of assessing the sources and oxidation pathways of atmospheric NOx (NO+NO₂). In the present study, daily PM₂.₅ samples were collected in the megacity of Beijing, China during the winter of 2017–2018, and this new approach was used to reveal the origin and oxidation pathways of atmospheric NOx. Specifically, the potential of field δ¹⁵N–NO₃⁻ signatures for determining the NOx oxidation chemistry was explored. Positive correlations between δ¹⁸O–NO₃⁻ and δ¹⁵N–NO₃⁻ were observed (with R² between 0.51 and 0.66, p < 0.01), and the underlying environmental significance was discussed. The results showed that the pathway-specific contributions to NO₃⁻ formation were approximately 45.3% from the OH pathway, 46.5% from N₂O₅ hydrolysis, and 8.2% from the NO₃+HC channel based on the δ¹⁸O-δ¹⁵N space of NO₃⁻. The overall nitrogen isotopic fractionation factor (εN) from NOx to NO₃⁻ on a daily scale, under winter conditions, was approximately +16.1‰±1.8‰ (consistent with previous reports). Two independent approaches were used to simulate the daily and monthly ambient NOx mixtures (δ¹⁵N-NOx), respectively. Results indicated that the monthly mean values of δ¹⁵N-NOx compared well based on the two approaches, with values of −5.5‰ ± 2.6‰, −2.7‰ ± 1.9‰, and −3.2‰ ± 2.2‰ for November, December, and January (2017–2018), respectively. The uncertainty was in the order of 5%, 5‰ and 5.2‰ for the pathway-specific contributions, the εN, and δ¹⁵N-NOx, respectively. Results also indicated that vehicular exhaust was the key contributor to the wintertime atmospheric NOx in Beijing (2017–2018). Our advanced isotopic perspective will support the future assessment of the origin and oxidation of urban atmospheric NOx.

Metal release into the environment from anthropogenic activities may endanger ecosystems and human health. However, identifying and quantifying anthropogenic metal bioaccumulation in organisms remain a challenging task. In this work, we assess Cu isotopes in Pacific oysters (C. gigas) as a new tool for monitoring anthropogenic Cu bioaccumulation into marine environments. Arcachon Bay was taken as a natural laboratory due to its increasing contamination by Cu, and its relevance as a prominent shellfish production area. Here, we transplanted 18-month old oysters reared in an oceanic neighbor area into two Arcachon Bay mariculture sites under different exposure levels to continental Cu inputs. At the end of their 12-month long transplantation period, the oysters’ Cu body burdens had increased, and was shifted toward more positive δ⁶⁵Cu values. The gradient of Cu isotope compositions observed for oysters sampling stations was consistent with relative geographic distance and exposure intensities to unknown continental Cu sources. A binary isotope mixing model based on experimental data allowed to estimate the Cu continental fraction bioaccumulated in the transplanted oysters. The positive δ⁶⁵Cu values and high bioaccumulated levels of Cu in transplanted oysters support that continental emissions are dominantly anthropogenic. However, identifying specific pollutant coastal source remained unelucidated mostly due to their broader and overlapping isotope signatures and potential post-depositional Cu isotope fractionation processes. Further investigations on isotope fractionation of Cu-based compounds in an aqueous medium may improve Cu source discrimination. Thus, using Cu as an example, this work combines for the first time a well-known caged bivalve approach with metal stable isotope techniques for monitoring and quantifying the bioaccumulation of anthropogenic metal into marine environments. Also, it states the main challenges to pinpoint specific coastal anthropogenic sources utilizing this approach and provides the perspectives for further studies to overcome them.

Hg accumulation in marine organisms depends strongly on in situ water or sediment biogeochemistry and levels of Hg pollution. To predict the rates of Hg exposure in human communities, it is important to understand Hg assimilation and processing within commercially harvested marine fish, like the European seabass Dicentrarchus labrax. Previously, values of Δ¹⁹⁹Hg and δ²⁰²Hg in muscle tissue successfully discriminated between seven populations of European seabass. In the present study, a multi-tissue approach was developed to assess the underlying processes behind such discrimination.We determined total Hg content (THg), the proportion of monomethyl-Hg (%MeHg), and Hg isotopic composition (e.g. Δ¹⁹⁹Hg and δ²⁰²Hg) in seabass liver. We compared this to the previously published data on muscle tissue and local anthropogenic Hg inputs.The first important finding of this study showed an increase of both %MeHg and δ²⁰²Hg values in muscle compared to liver in all populations, suggesting the occurrence of internal MeHg demethylation in seabass. This is the first evidence of such a process occurring in this species. Values for mass-dependent (MDF, δ²⁰²Hg) and mass-independent (MIF, Δ¹⁹⁹Hg) isotopic fractionation in liver and muscle accorded with data observed in estuarine fish (MDF, 0–1‰ and MIF, 0–0.7‰). Black Sea seabass stood out from other regions, presenting higher MIF values (≈1.5‰) in muscle and very low MDF (≈-1‰) in liver. This second finding suggests that under low Hg bioaccumulation, Hg isotopic composition may allow the detection of a shift in the habitat use of juvenile fish, such as for first-year Black Sea seabass.Our study supports the multi-tissue approach as a valid tool for refining the analysis of Hg sourcing and metabolism in a marine fish. The study’s major outcome indicates that Hg levels of pollution and fish foraging location are the main factors influencing Hg species accumulation and isotopic fractionation in the organisms.

Trace elements and Hg isotopic composition were investigated in mineralized rocks, barren rocks, and mineral soils in the Xianfeng prospect, a shallow buried epithermal gold deposit in northeastern China, to understand whether this deposit has left a diagnostic geochemical fingerprint to its weathered horizon. All the rocks and soils display congruent patterns for immobile elements (large ion lithophile elements, high field strength elements, and rare earth elements), which reflect the subduction-related tectonic setting. Both mineralized rocks and soils showed common enrichment of elemental suite As–Ag–Sb–Hg, suggesting that the Xianfeng gold deposit has released these elements into its weathered horizon. Similar mercury isotopic composition was observed between mineralized rocks (δ²⁰²Hg: −0.21 ± 0.70‰; Δ¹⁹⁹Hg: −0.02 ± 0.12‰; 2SD) and barren rocks (δ²⁰²Hg: −0.46 ± 0.48‰; Δ¹⁹⁹Hg: 0.00 ± 0.10‰; 2SD), suggesting that mercury in the Xianfeng deposit is mainly derived from the magmatic rocks. Mineralized soils (δ²⁰²Hg: −0.44 ± 0.60‰; −0.03 ± 0.14‰; 2SD) and barren soils (δ²⁰²Hg: −0.54 ± 0.68‰; Δ¹⁹⁹Hg: −0.05 ± 0.14‰; 2SD) displayed congruent Hg isotopic signals to the underlying rocks, suggesting limited Hg isotope fractionation during the release of Hg from ore deposit to soils via weathering. This study reveals evidence of a simple and direct geochemical link between this shallow buried hydrothermal deposit and its weathered horizon, and highlights that the weathering of shallow-buried hydrothermal gold deposits can release a substantial amount of heavy metals (e.g. Hg, As and Sb) to surface soil.

Tributyl phosphate (TBP) is recognised as a global environmental contaminant because of its wide use in floatation reagents, nuclear fuel reprocessing and plasticisers. This contaminant is hardly degraded by hydrolysis in the environment due to its special physicochemical properties. In this study, one TBP-degrading strain was isolated from TBP-contaminated abandoned mine tailings, and 16S rRNA identification revealed that the strain belonged to the genus Sphingomonas. Results validated that the strain could utilise TBP as the sole carbon source, and vitamin was not the essential factor for its growth. Liquid chromatography time-of-flight mass spectrometry analysis identified di-n-butyl phosphate (DnBP) and mono-n-butyl phosphate (MnBP) as the intermediate metabolites for TBP biodegradation. No obvious change in carbon and hydrogen isotope composition was observed in biodegradation processes (cell suspension and crude extract degradation), which indicated that the first irreversible bond cleavage did not involve carbon or hydrogen. Hence, the TBP degradation scheme by Sphingomonas sp. proposed that the first irreversible step of TBP transferred to DnBP would lead to PO bond cleavage. This study combined the identification of products and isotope fractionation in substrates to investigate the transformation mechanism, thereby providing an eco-friendly and cost-effective way for the in situ bioremediation of TBP-contaminated sites by the isolated TBP degradation strain.

The use of Ni and Cu isotopes for tracing contamination sources in the environment remains a challenging task due to the limited information about the influence of various biogeochemical processes influencing stable isotope fractionation. This work focuses on a relatively simple system in north-east Norway with two possible endmembers (smelter-bedrock) and various environmental samples (snow, soil, lichens, PM10). In general, the whole area is enriched in heavy Ni and Cu isotopes highlighting the impact of the smelting activity. However, the environmental samples exhibit a large range of δ⁶⁰Ni (−0.01 ± 0.03‰ to 1.71 ± 0.02‰) and δ⁶⁵Cu (−0.06 ± 0.06‰ to −3.94 ± 0.3‰) values which exceeds the range of δ⁶⁰Ni and δ⁶⁵Cu values determined in the smelter, i.e. in feeding material and slag (δ⁶⁰Ni from 0.56 ± 0.06‰ to 1.00 ± 0.06‰ and δ⁶⁵Cu from −1.67 ± 0.04‰ to −1.68 ± 0.15‰). The shift toward heavier Ni and Cu δ values was the most significant in organic rich topsoil samples in the case of Ni (δ⁶⁰Ni up to 1.71 ± 0.02‰) and in lichens and snow in the case of Cu (δ⁶⁵Cu up to −0.06 ± 0.06‰ and −0.24 ± 0.04‰, respectively). These data suggest an important biological and biochemical fractionation (microorganisms and/or metal uptake by higher plants, organo-complexation etc.) of Ni and Cu isotopes, which should be quantified separately for each process and taken into account when using the stable isotopes for tracing contamination in the environment.