PROCESOS DE ATENUACION DE CONTAMINANTES CONVENCIONALES Y EMERGENTES PROCEDENTES DE FUENTES AGRICOLAS E INDUSTRIALES EN AGUAS
CGL2017-87216-C4-1-R
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Nombre agencia financiadora Agencia Estatal de Investigación
Acrónimo agencia financiadora AEI
Programa Programa Estatal de I+D+i Orientada a los Retos de la Sociedad
Subprograma Programa Estatal de I+D+i Orientada a los Retos de la Sociedad
Convocatoria Retos Investigación: Proyectos I+D+i
Año convocatoria 2017
Unidad de gestión Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016
Centro beneficiario UNIVERSIDAD DE BARCELONA
Identificador persistente http://dx.doi.org/10.13039/501100011033
Publicaciones
Resultados totales (Incluyendo duplicados): 10
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Multi-method assessment of the intrinsic biodegradation potential of an aquifer contaminated with chlorinated ethenes at an industrial area in Barcelona (Spain)
Dipòsit Digital de Documents de la UAB
- Blázquez Pallí, Natàlia|||0000-0001-5242-5818
- Rosell, Mònica|||0000-0003-1563-8595
- Varias, Joan
- Bosch, Marçal|||0000-0001-5829-7829
- Soler, Albert
- Vicent i Huguet, Teresa|||0000-0002-6547-7358
- Marco Urrea, Ernest|||0000-0002-8033-6553
The bioremediation potential of an aquifer contaminated with tetrachloroethene (PCE) was assessed by combining hydrogeochemical data of the site, microcosm studies, metabolites concentrations, compound specific-stable carbon isotope analysis and the identification of selected reductive dechlorination biomarker genes. The characterization of the site through 10 monitoring wells evidenced that leaked PCE was transformed to TCE and cis-DCE via hydrogenolysis. Carbon isotopic mass balance of chlorinated ethenes pointed to two distinct sources of contamination and discarded relevant alternate degradation pathways in the aquifer. Application of specific-genus primers targeting Dehalococcoides mccartyi species and the vinyl chloride-to-ethene reductive dehalogenase vcrA indicated the presence of autochthonous bacteria capable of the complete dechlorination of PCE. The observed cis-DCE stall was consistent with the aquifer geochemistry (positive redox potentials; presence of dissolved oxygen, nitrate, and sulphate; absence of ferrous iron), which was thermodynamically favourable to dechlorinate highly chlorinated ethenes but required lower redox potentials to evolve beyond cis-DCE to the innocuous end product ethene. Accordingly, the addition of lactate or a mixture of ethanol plus methanol as electron donor sources in parallel field-derived anoxic microcosms accelerated dechlorination of PCE and passed cis-DCE up to ethene, unlike the controls (without amendments, representative of field natural attenuation). Lactate fermentation produced acetate at near-stoichiometric amounts. The array of techniques used in this study provided complementary lines of evidence to suggest that enhanced anaerobic bioremediation using lactate as electron donor source is a feasible strategy to successfully decontaminate this site.
Dual carbon-chlorine isotope fractionation during dichloroelimination od 1,1,2-trichloroethane by an enrichment culture containing Dehalogenimonas sp
Dipòsit Digital de Documents de la UAB
- Rosell, Mònica|||0000-0003-1563-8595
- Palau Capdevila, Jordi|||0000-0001-9492-7306
- Mortan, Siti Hatijah|||0000-0003-4968-1508
- Caminal i Saperas, Glòria|||0000-0001-9646-6099
- Soler, Albert
- Shouakar Stash, Orfan
- Marco Urrea, Ernest|||0000-0002-8033-6553
Chlorinated ethanes are frequent groundwater contaminants but compound specific isotope analysis (CSIA) has been scarcely applied to investigate their degradation pathways. In this study, dual carbon and chlorine isotope fractionation was used to investigate for the first time the anoxic biodegradation of 1,1,2-trichloroethane (1,1,2-TCA) using a Dehalogenimonas-containing culture. The isotopic fractionation values obtained for the biodegradation of 1,1,2-TCA were ɛC = −6.9 ± 0.4‰ and ɛCl = −2.7 ± 0.3‰. The detection of vinyl chloride (VC) as unique byproduct and a closed carbon isotopic mass balance corroborated that dichloroelimination was the degradation pathway used by this strain. Combining the values of δ13C and δ37Cl resulted in a dual element C-Cl isotope slope of Λ = 2.5 ± 0.2‰. Investigation of the apparent kinetic isotope effects (AKIEs) expected for cleavage of a CCl bond showed an important masking of the intrinsic isotope fractionation. Theoretical calculation of Λ suggested that dichloroelimination of 1,1,2-TCA was taking place via simultaneous cleavage of two CCl bonds (concerted reaction mechanism). The isotope data obtained in this study can be useful to monitor natural attenuation of 1,1,2-TCA via dichloroelimination and provide insights into the source and fate of VC in contaminated groundwaters.
Use of dual element isotope analysis and microcosm studies to determine the origin and potential anaerobic biodegradation of dichloromethane in two multi-contaminated aquifers
Dipòsit Digital de Documents de la UAB
- Blázquez Pallí, Natàlia|||0000-0001-5242-5818
- Shouakar Stash, Orfan
- Palau Capdevila, Jordi|||0000-0001-9492-7306
- Trueba-Santiso, Alba|||0000-0001-9730-7321
- Varias, Joan
- Bosch, Marçal|||0000-0001-5829-7829
- Soler, Albert
- Vicent i Huguet, Teresa|||0000-0002-6547-7358
- Marco Urrea, Ernest|||0000-0002-8033-6553
- Rosell, Mònica|||0000-0003-1563-8595
Many aquifers around the world are impacted by toxic chlorinated methanes derived from industrial processes due to accidental spills. Frequently, these contaminants co-occur with chlorinated ethenes and/or chlorinated benzenes in groundwater, forming complex mixtures that become very difficult to remediate. In this study, a multi-method approach was used to provide lines of evidence of natural attenuation processes and potential setbacks in the implementation of bioremediation strategies in multi-contaminated aquifers. First, this study determined i) the carbon and chlorine isotopic compositions (δ¹³C, δ³⁷Cl) of several commercial pure phase chlorinated compounds, and ii) the chlorine isotopic fractionation (εCl = −5.2 ± 0.6‰) and the dual CCl isotope correlation (ΛC/Cl = 5.9 ± 0.3) during dichloromethane (DCM) degradation by a Dehalobacterium-containing culture. Such data provide valuable information for practitioners to support the interpretation of stable isotope analyses derived from polluted sites. Second, the bioremediation potential of two industrial sites contaminated with a mixture of organic pollutants (mainly DCM, chloroform (CF), trichloroethene (TCE), and mono-chlorobenzene (MCB)) was evaluated. Hydrochemistry, dual (CCl) isotope analyses, laboratory microcosms, and microbiological data were used to investigate the origin, fate and biodegradation potential of chlorinated methanes. At Site 1, δ¹³C and δ³⁷Cl compositions from field samples were consistent with laboratory microcosms, which showed complete degradation of CF, DCM and TCE, while MCB remained. Identification of Dehalobacter sp. in CF-enriched microcosms further supported the biodegradation capability of the aquifer to remediate chlorinated methanes. At Site 2, hydrochemistry and δ¹³C and δ³⁷Cl compositions from field samples suggested little DCM, CF and TCE transformation; however, laboratory microcosms evidenced that their degradation was severely inhibited, probably by co-contamination. A dual CCl isotopic assessment using results from this study and reference values from the literature allowed to determine the extent of degradation and elucidated the origin of chlorinated methanes.
Integrative isotopic and molecular approach for the diagnosis and implementation of an efficient in-situ enhanced biological reductive dechlorination of chlorinated ethenes
Dipòsit Digital de Documents de la UAB
- Blázquez Pallí, Natàlia|||0000-0001-5242-5818
- Rosell, Mònica|||0000-0003-1563-8595
- Varias, Joan
- Boscn, Marcal
- Soler, Albert
- Vicent i Huguet, Teresa|||0000-0002-6547-7358
- Marco Urrea, Ernest|||0000-0002-8033-6553
Based on the previously observed intrinsic bioremediation potential of a site originally contaminated with perchloroethene (PCE), field-derived lactate-amended microcosms were performed to test different lactate isomers and concentrations, and find clearer isotopic and molecular parameters proving the feasibility of an in-situ enhanced reductive dechlorination (ERD) from PCE-to-ethene (ETH). According to these laboratory results, which confirmed the presence of Dehalococcoides sp. and the vcrA gene, an in-situ ERD pilot test consisting of a single injection of lactate in a monitoring well was performed and monitored for 190 days. The parameters used to follow the performance of the ERD comprised the analysis of i) hydrochemistry, including redox potential (Eh), and the concentrations of redox sensitive species, chlorinated ethenes (CEs), lactate, and acetate; ii) stable isotope composition of carbon of CEs, and sulphur and oxygen of sulphate; and iii) 16S rRNA gene sequencing from groundwater samples. Thus, it was proved that the injection of lactate promoted sulphate-reducing conditions, with the subsequent decrease in Eh, which allowed for the full reductive dechlorination of PCE to ETH in the injection well. The biodegradation of CEs was also confirmed by the enrichment in 13C and carbon isotopic mass balances. The metagenomic results evidenced the shift in the composition of the microbial population towards the predominance of fermentative bacteria. Given the success of the in-situ pilot test, a full-scale ERD with lactate was then implemented at the site. After one year of treatment, PCE and trichloroethene were mostly depleted, whereas vinyl chloride (VC) and ETH were the predominant metabolites. Most importantly, the shift of the carbon isotopic mass balances towards more positive values confirmed the complete reductive dechlorination, including the VC-to-ETH reaction step. The combination of techniques used here provides complementary lines of evidence for the diagnosis of the intrinsic biodegradation potential of a polluted site, but also to monitor the progress, identify potential difficulties, and evaluate the success of ERD at the field scale.
Trichloromethane dechlorination by a novel Dehalobacter sp. strain 8M reveals a third contrasting C and Cl isotope fractionation pattern within this genus
Dipòsit Digital de Documents de la UAB
- Soder-Walz, Jesica M|||0000-0002-5253-9898
- Torrentó, Clara|||0000-0003-1480-2744
- Algora, Camelia
- Wasmund, Kenneth|||0000-0001-6706-7291
- Cortés Garmendia, M. Pilar|||0000-0002-5924-0653
- Soler, Albert
- Vicent i Huguet, Teresa|||0000-0002-6547-7358
- Rosell, Mònica|||0000-0003-1563-8595
- Marco Urrea, Ernest|||0000-0002-8033-6553
Trichloromethane (TCM) is a pollutant frequently detected in contaminated aquifers, and only four bacterial strains are known to respire it. Here, we obtained a novel Dehalobacter strain capable of transforming TCM to dichloromethane, which was denominated Dehalobacter sp. strain 8M. Besides TCM, strain 8M also completely transformed 1,1,2-trichloroethane to vinyl chloride and 1,2-dichloroethane. Quantitative PCR analysis for the 16S rRNA genes confirmed growth of Dehalobacter with TCM and 1,1,2-trichloroethane as electron acceptors. Carbon and chlorine isotope fractionation during TCM transformation was studied in cultured cells and in enzymatic assays with cell suspensions and crude protein extracts. TCM transformation in the three studied systems resulted in small but significant carbon (εC = −2.7 ± 0.1‰ for respiring cells, −3.1 ± 0.1‰ for cell suspensions, and − 4.1 ± 0.5‰ for crude protein extracts) and chlorine (εCl = −0.9 ± 0.1‰, −1.1 ± 0.1‰, and − 1.2 ± 0.2‰, respectively) isotope fractionation. A characteristic and consistent dual Csingle bondCl isotope fractionation pattern was observed for the three systems (combined ΛC/Cl = 2.8 ± 0.3). This ΛC/Cl differed significantly from previously reported values for anaerobic dechlorination of TCM by the corrinoid cofactor vitamin B12 and other Dehalobacter strains. These findings widen our knowledge on the existence of different enzyme binding mechanisms underlying TCM-dechlorination within the genus Dehalobacter and demonstrates that dual isotope analysis could be a feasible tool to differentiate TCM degraders at field studies.
Dual C-Cl isotope fractionation offers potential to assess biodegradation of 1,2-dichloropropane and 1,2,3-trichloropropane by Dehalogenimonas cultures
Dipòsit Digital de Documents de la UAB
- Trueba-Santiso, Alba|||0000-0001-9730-7321
- Torrentó, Clara|||0000-0003-1480-2744
- Soder-Walz, Jesica M.|||0000-0002-5253-9898
- Fernandez Verdejo, David Juan|||0000-0002-3234-4006
- Rosell, Mònica|||0000-0003-1563-8595
- Marco Urrea, Ernest|||0000-0002-8033-6553
1,2-dichloropropane (1,2-DCP) and 1,2,3-trichloropropane (1,2,3-TCP) are hazardous chemicals frequently detected in groundwater near agricultural zones due to their historical use in chlorinated fumigant formulations. In this study, we show that the organohalide-respiring bacterium Dehalogenimonas alkenigignens strain BRE15 M can grow during the dihaloelimination of 1,2-DCP and 1,2,3-TCP to propene and allyl chloride, respectively. Our work also provides the first application of dual isotope approach to investigate the anaerobic reductive dechlorination of 1,2-DCP and 1,2,3-TCP. Stable carbon and chlorine isotope fractionation values for 1,2-DCP (ƐC = −13.6 ± 1.4 ‰ and ƐCl = −27.4 ± 5.2 ‰) and 1,2,3-TCP (ƐC = −3.8 ± 0.6 ‰ and ƐCl = −0.8 ± 0.5 ‰) were obtained resulting in distinct dual isotope slopes (Λ12DCP = 0.5 ± 0.1, Λ123TCP = 4 ± 2). However direct comparison of ΛC-Cl among different substrates is not possible and investigation of the C and Cl apparent kinetic isotope effects lead to the hypothesis that concerted dichloroelimination mechanism is more likely for both compounds. In fact, whole cell activity assays using cells suspensions of the Dehalogenimonas-containing culture grown with 1,2-DCP and methyl viologen as electron donor suggest that the same set of reductive dehalogenases was involved in the transformation of 1,2-DCP and 1,2,3-TCP. This study opens the door to the application of isotope techniques for evaluating biodegradation of 1,2-DCP and 1,2,3-TCP, which often co-occur in groundwaters near agricultural fields.
DOI: https://ddd.uab.cat/record/291860, https://dx.doi.org/urn:doi:10.1016/j.chemosphere.2024.142170
A multi-isotopic approach to investigate the influence of land use on nitrate removal in a highly saline lake-aquifer system
Digital.CSIC. Repositorio Institucional del CSIC
- Valiente, Nicolas
- Carrey, Raúl
- Otero, Neus
- Soler, Albert
- Sanz, David
- Muñoz-Martín, A.
- Jirsa, Franz
- Wanek, W.
- Gómez-Alday, Juan José
Endorheic or closed drainage basins in arid and semi-arid regions are vulnerable to pollution. Nonetheless, in the freshwater-saltwater interface of endorheic saline lakes, oxidation-reduction (redox) reactions can attenuate pollutants such as nitrate (NO3−). This study traces the ways of nitrogen (N) removal in the Pétrola lake-aquifer system (central Spain), an endorheic basin contaminated with NO3− (up to 99.2 mg/L in groundwater). This basin was declared vulnerable to NO3− pollution in 1998 due to the high anthropogenic pressures (mainly agriculture and wastewaters). Hydrochemical, multi-isotopic (δ18ONO3, δ15NNO3, δ13CDIC, δ18OH2O, and δ2HH2O) and geophysical techniques (electrical resistivity tomography) were applied to identify the main redox processes at the freshwater-saltwater interface. The results showed that the geometry of this interface is influenced by land use, causing spatial variability of nitrogen biogeochemical processes over the basin. In the underlying aquifer, NO3− showed an average concentration of 38.5 mg/L (n = 73) and was mainly derived from agricultural inputs. Natural attenuation of NO3− was observed in dryland farming areas (up to 72%) and in irrigation areas (up to 66%). In the Pétrola Lake, mineralization and organic matter degradation in lake sediment play an important role in NO3− reduction. Our findings are a major step forward in understanding freshwater-saltwater interfaces as reactive zones for NO3− attenuation. We further emphasize the importance of including a land use perspective when studying water quality-environmental relationships in hydrogeological systems dominated by density-driven circulation., his work was financed by a PhD grant (BES-2012-052256) from the Spanish government, the PEIC-2014-004-P project from the Castilla–La Mancha regional government, the projects CICYT CGL2014-57215-C4-1-R, CGL2017-87216-C4-1-R and CGL2017-87216-C4-2-R from the Spanish Ministry of Economy and AEU/FEDER UE, and the project 2017SGR1733 from the Generalitat de Catalunya., Peer reviewed
Geochemical and isotopic study of abiotic nitrite reduction coupled to biologically produced Fe(II) oxidation in marine environments
Digital.CSIC. Repositorio Institucional del CSIC
- Benaiges-Fernández, Robert
- Offeddu, Francesco G.
- Margalef-Martí, R.
- Palau, Jordi
- Urmeneta, Jordi
- Carrey, Raúl
- Otero, Neus
- Cama, Jordi
Estuarine sediments are often characterized by abundant iron oxides, organic matter, and anthropogenic nitrogen compounds (e.g., nitrate and nitrite). Anoxic dissimilatory iron reducing bacteria (e.g., Shewanella loihica) are ubiquitous in these environments where they can catalyze the reduction of Fe(III) (oxyhydr)oxides, thereby releasing aqueous Fe(II). The biologically produced Fe(II) can later reduce nitrite to form nitrous oxide. The effect on nitrite reduction by both biologically produced and artificially amended Fe(II) was examined experimentally. Ferrihydrite was reduced by Shewanella loihica in a batch reaction with an anoxic synthetic sea water medium. Some of the Fe(II) released by S. loihica adsorbed onto ferrihydrite, which was involved in the transformation of ferrihydrite to magnetite. In a second set of experiments with identical medium, no microorganism was present, instead, Fe(II) was amended. The amount of solid-bound Fe(II) in the experiments with bioproduced Fe(II) increased the rate of abiotic NO2− reduction with respect to that with synthetic Fe(II), yielding half-lives of 0.07 and 0.47 d, respectively. The δ18O and δ15N of NO2− was measured through time for both the abiotic and innoculated experiments. The ratio of ε18O/ε15N was 0.6 for the abiotic experiments and 3.1 when NO2− was reduced by S. loihica, thus indicating two different mechanisms for the NO2− reduction. Notably, there is a wide range of the ε18O/ε15N values in the literature for abiotic and biotic NO2− reduction, as such, the use of this ratio to distinguish between reduction mechanisms in natural systems should be taken with caution. Therefore, we suggest an additional constraint to identify the mechanisms (i.e. abiotic/biotic) controlling NO2− reduction in natural settings through the correlation of δ15N-NO2- and the aqueous Fe(II) concentration., This study was supported by projects CGL2017-87216-C4-1-R, CGL2017-82331-R and CEX2018-000794-S funded by the Spanish Ministry of Science and Innovation and AEI/FEDER funded by the European Union, and by MAG (2017 SGR 1733) financed by the Catalan Government. R. Margalef-Marti wishes to thank the Spanish Government for the Ph.D. grant BES-2015-072882. The authors are indebted to Jordi Bellés (IDAEA-CSIC), Natàlia Moreno (IDAEA-CSIC) and Xavier Alcové (SCTT-Barcelona University) for laboratory assistance and XRD analyses, respectively. The isotopic analyses were prepared at the MAiMA-UB research group laboratory and analyzed at the scientific and technical services of Barcelona University (CCiT-UB). We acknowledge Max Giannetta for his scientific discussions during the manuscript elaboration. We also wish to thank the Editor and three anonymous reviewers for their constructive comments that have improved the quality of the paper., Peer reviewed
Enhancing Nitrate Removal With Industrial Wine Residue: Insights From Laboratory Batch and Column Experiments Using Chemical, Isotopic and Numerical Modeling Tools
Digital.CSIC. Repositorio Institucional del CSIC
- Abu, Alex
- Carrey, Raúl
- Navarro-Ciurana, Didac
- Margalef-Martí, R.
- Soler, Albert
- Otero, Neus
- Causapé Valenzuela, Jesús Antonio
- Domènech, Cristina
[EN] Agricultural run-off exposes recipient water bodies to nitrate (NO3−) pollution. Biological denitrification is a suitable method for removing NO3− in water resources that can be induced by the use of industrial organic liquid waste as an electron donor source. In light of this, batch and column laboratory experiments were performed to assess the potential of two industrial wine residues (lías and vínico) to induce biological denitrification of NO3− contaminated water from a constructed wetland and to evaluate the efficiency of these treatments using chemical and isotopic tools. In batch experiments (performed at a C/N ratio of 1.25), vínico was not efficient enough in removing N species, attenuating only 35% NO3− and was not used in column experiments. In similar experimental conditions, lías completely removed N species from water in both batch and column experiments. The calculated isotope fractionation (ε15NNO3 and ε18ONO3) was the same in both batch and column experiments biostimulated with lías and differed from those for vínico. The isotopic data confirmed that denitrification was the principal NO3− attenuation pathway in all the experiments. The isotopic fractionation can be later applied to field studies to quantify the efficiency of biologically enhanced denitrification. A numerical geochemical model that accounts for the changes in nitrate, nitrite concentration and isotopic composition due to the degradation of lías and vínico, including transport in the case of the column experiment, was performed to simulate the experimental results and can be up-scaled in field treatments., This publication is part of the Grant PACE-ISOTEC (CGL2017-87216-C4-1-R) funded by MICIU/AEI/10.13039/501100011033 and by “ERDF A way of making Europe,” the Grant REMEDIATE (TED2021-131005B-C31) funded by MICIU/AEI/10.13039/501100011033 and by “European Union NextGenerationEU/PRTR,” the Grant NPP-SOL (PCI2023-143359) funded by MICIU/AEI/10.13039/501100011033 and co-funded by the European Union under the “PRIMA2022 Programme-Section2” and the project AgroSOS (PID2019-108057RB-I00) funded by MICIU/AEI/10.13039/501100011033 and by “ERDF/EU”. It has also been funded by Generalitat de Catalunya through the Consolidated Research Group MAGH (2021-SGR-00308). The authors are thankful to the CCiT of the Universitat de Barcelona for the analytical support. Abu Alex would also like to thank the Agency for Management of University and Research Grants of the Generalitat de Catalunya and Fundació Bosch i Gimpera de la Universitat de Barcelona for the PhD Grant (2019 FI_B 01059) and the L’Institut de Recerca de l’Aigua (IdRA)-Universitat de Barcelona for their support., Peer reviewed
DOI: http://hdl.handle.net/10261/374190, https://api.elsevier.com/content/abstract/scopus_id/85193042603
Feasibility of using rural waste products to increase the denitrification efficiency in a surface flow constructed wetland
Zaguán. Repositorio Digital de la Universidad de Zaragoza
- Margalef-Marti, R.
- Carrey, R.
- Merchán, D.
- Soler, A.
- Causapé, J.
- Otero, N.
A surface flow constructed wetland (CW) was set in the Lerma gully to decrease nitrate (NO3 -) pollution from agricultural runoff water. The water flow rate and NO3 - concentration were monitored at the inlet and the outlet, and sampling campaigns were performed which consisted of collecting six water samples along the CW flow line. After two years of operation, the NO3 - attenuation was limited at a flow rate of ~2.5 L/s and became negligible at ~5.5 L/s. The present work aimed to assess the feasibility of using rural waste products (wheat hay, corn stubble, and animal compost) to induce denitrification in the CW, to assess the effect of temperature on this process, and to trace the efficiency of the treatment by using isotopic tools. In the first stage, microcosm experiments were performed. Afterwards, the selected waste material was applied in the CW, and the treatment efficiency was evaluated by means of a chemical and isotopic characterization and using the isotopic fractionation (e) values calculated from laboratory experiments to avoid field-scale interference. The microcosms results showed that the stubble was the most appropriate material for application in the CW, but the denitrification rate was found to decrease with temperature. In the CW, biostimulation in autumn-winter promoted NO3 - attenuation between two weeks and one month (a reduction in NO3 - between 1.2 and 1.5 mM was achieved). After the biostimulation in spring-summer, the attenuation was maintained for approximately three months (NO3 - reduction between 0.1 and 1.5 mM). The e15NNO3/N2 and e18ONO3/N2 values obtained from the laboratory experiments allowed to estimate the induced denitrification percentage. At an approximate average flow rate of 16 L/s, at least 60% of NO3 - attenuation was achieved in the CW. The field samples exhibited a slope of 1.0 for d18O-NO3 - versus d15N-NO3 -, similar to those of the laboratory experiments (0.9–1.2). Plant uptake seemed to play a minor role in NO3 - attenuation in the CW. Hence, the application of stubble in the CW allowed the removal of large amounts of NO3 - from the Lerma gully, especially when applied during the warm months, but its efficacy was limited to a short time period (up to three months). © 2019 Elsevier B.V.