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damia barcelo culleres 

Coordinator: Damià Barceló Culleres


Projects  Partner and Institution:
Damià Barceló (CSIC)
Peter Schröder (HMGU)
Serge Chiron (UM-HSM)
Trine Egen (NIBIO)
Fabrice Martin-Laurent (INRA

Key words: WWTP, emerging contaminants, (bio)sensors, electrochemical and biological treatment processes, ecotoxicological assessment, sustainable water reuse and safety, LCA


With growing demand for good quality water in agriculture, the use of reclaimed water has become an extended practice in water-shortage regions. The use of treated wastewater in agriculture can complement natural fresh water resources if needed for crop irrigation while contributing to nutrient recycling. However, it represents a primary source of water-borne contaminants escaping removal during to wastewater treatment processes. Furthermore, in agriculture a major concern is the direct application of pesticides which manifests in groundwater contamination, surface run-off and plant uptake. Concerns about water resources are intensified by differing levels of quality and particularly by environmental soundness. The usage patterns of water among European countries differ strongly. Northern European countries mostly rely on natural precipitation while many South European countries are facing difficulties in meeting the water demand of crops. In some European countries the use of treated wastewater has been implemented for irrigation when natural freshwater sources are scarce. Hence, the AWARE project aims to investigate the fate of pesticides applied directly and wastewater-borne contaminants from irrigation with treated wastewater in soil/plants evaluating their presence, biodegradation/metabolism and environmental risks in agricultural fields.

These goals have been addressed by:

1) evaluating the uptake and metabolism of 14 relevant wastewater-derived pollutants in cultivated lettuce and radish under controlled conditions and under real conditions (CSIC, Spain; HMGU, Germany; UM, France). First, we developed analytical methods for determining the 14 compounds in crop samples and second the plants have been cultivated under controlled conditions and in field studies. Next step will be to apply these methods to analyze the selected compounds in the cultivated crops.

2) We also investigated the degradation of 14 compounds and five pesticides in soil. The capacity of the ascomycete fungus genus Trichoderma (Trichoderma species are highly present in nearly all soil types where they grow and live in symbiotic relationship with plants) for 14 compounds biodegradation has been investigated in UM (France) and they have shown high capacity in degrading selected compounds.  Pesticide removal was investigated in a microcosm horizontal subsurface.

3) Estimating the ecotoxicological impact of 14 contaminants on soil microbial diversity using mainly DNA based technologies which allowed the estimation of the abundance diversity and activity of the microbial activity as well as the use of earthworm’s carboxylesterases for the assessment of the 14 compounds exposure (CSIC, Spain; INRA, France). The samples are now being processed for their chemical analysis.

Finally, we will integrate all the results to develop a decision-support system that will be co-developed with stakeholders from different countries reflecting different socioenvironmental settings from the northern and southern European countries. Our training and knowledge exchange activities with farmers, growers, and agronomists, which will build the capacity of these groups to apply novel irrigation strategies more effectively.

Project structure:

Implementation: Overall coherence and effectiveness of the work plan. The methodological approach of the current proposal is divided into the following WPs (see table 1 and gant chart). WP1. Task 1. Selection of soil/filter-system to be investigate in constructed wetlands (CWs) and in an advanced riparian strip zone will be performed after a small scale laboratory pre-evaluation of sorption capacity of cost-and-practical-relevant filter materials (M1.2). The most promising soil/filter system will be investigated in controlled experiment systems located at UFZ Leipzig (Collaborator); a constructed wetland (CW) facility with three Planted Fixed Bed Reactors (PFR, system for investigating rhizosphere processes) which can be taken out of the CW and transferred to a phytotechnicum or studying processes in detail (M1.1). Parent compounds and important metabolites will be analyzed for, and experiments will be designed in order to separate retention and degradation, and also the effect of reductive zones in a CW. Improved sorption and stimulation of degradation pesticides by including filter material with high affinity to hydrophilic compounds would be an innovative improvement even it still will be a low-technology solution. Task 2. For quantifying turnover and NER-formation of imidacloprid, we will use similar methodology as UFZ Leipzig apply; stable isotope labelled pesticide, abiotic and biotic incubation following OECD guidelines 307 and 308 (M1.4). Task 3. The degradation of selected compounds (sulfamethazole, diclofenac, carbamazepine, ofloxacin and iopromide) will be carried out in aqueous phase under aerobic conditions using T. harzianum strain T22 (D1.1). Kinetic results, the formation of potential transformation products and transformation pathways will be compared with those previously obtained with Trametes versicolor, a well-studied lignolytic fungus for pharmaceutical degradation. BTPs will be identified by LC-HRMS. Relevant BTPs will be searched in real soil/plant systems (see WP4) to confirm the implication of Trichoderma species in pharmaceutical attenuation processes in rizhosphere. Task 4. Determination of enantiomer ratios of climbazole and metoprolol on the basis of the chiral analytical methods developed for wastewater and sludge (D1.2). Data will be processed using the Rayleigh equation to investigate the relationship between the extend of climbazole and metoprolol biodegradation and their enantiomeric ratios and for their quantitative biodegradation assessment in soil. WP2. In this WP the capabilities of several plant species to metabolize different relevant drugs will be investigated. Plants have been selected for their widespread across the EU and their fast germination, growth and biomass production. The application of suspect screening on a HR-MS platform will demonstrate the power of this emerging analytical approach. By performing the sample analysis in a completely unbiased mode, the need for method development is minimized and the time spent for sample preparation is reduced. The acquisition of mass spectral data is software-guided in order to maximize the amount of information gathered in each run (M2.1). Together with the metabolism and kinetic data, a comprehensive picture of the fate of compounds under consideration will be obtained (M2.4). Molecular approaches will be used to pinpoint detoxification enzymes of special importance for the degradation of the selected drugs, and ways to enhance their activity will be explored. Critical concentrations of drugs leading to oxidative stress reactions in plants will be considered and passed to WP4 for completion of the risk assessment (D2.2). In order to obtain best accuracy, chemical analysis and enzymatic assays will at least be once done from the same samples prepared by one of the participating labs. Results will be of special importance for WP1 and 3 and will be transferred to these partners in due course. The use of bioassays together with advanced analytics will enable AWARE to give clear recommendations about actions to be taken with respect to suspect drugs, and biomass fractions (M2.3, D2.4). WP 3. For crop cultivation, rate of irrigation, frequency of irrigation, duration of irrigation, plant species, pre-harvest waiting periods will be compiled and tabulated (M3.1). Pharmaceuticals residues will be analyzed in irrigation water, soil, plant tissue and soil leachates by suspect screening approaches using HR-MS platforms (M3.4). For health risks, the estimated daily intake (EDI) value for each pharmaceutical and plant species combination will be compared to acceptable daily intake (ADI) values to establish a hazard quotient. In presence of a mixture of pharmaceuticals and their TPs in the edible tissue of crops, a conservative approach will be applied by summing all the calculated hazard quotients. For risk assessment of pharmaceutical accumulation in soil or leaching to groundwater, pharmaceuticals will be recovered following the soil profiles to investigate their movement through the soil horizons and amounts in leachates will be determined to investigate which compounds or TPs are likely to pose a risk to groundwater. These data obtained at environmentally concentrations will be used to calibrate existing reactive transfer models. For TWW reuse, environmental and health risk assessment will be assessed under field conditions (D3.3). Crop irrigation activities with treated wastewater will be performed at the experimental site of Murviel Lès Montpellier site which will be plotted with radish (Raphanus sativus), lettuce (Lactuca sativa) and zucchini (Curcubita pepo) because these crops have extensive root development at early growth stages, facilitating studies of plant uptake and studies of effects on plant crop. Rate of irrigation, frequency of irrigation, duration of irrigation, plant species, pre-harvest waiting periods will be compiled and tabulated because all these parameters are relevant for determining exposure scenarios. Pore water, soil and plant sample collections to be prepared and sent to all partners according to their analytical capacity and skills. UM will be in charge to collect and compile all the analytical results. For this purpose, NIBIO will benefit from The National Monitoring Program and these facilities and will also have access to other established monitoring systems with high relevance for studying pesticide retention in an advanced riparian strip zone (D3.2). WP 4. Main objectives of this WP are to assess the impact of pesticide and pharmaceuticals residues coming from wastewater irrigation on soil and plant. To reach this objective, the first step will consist in characterizing agronomically relevant scenario of exposure to pesticides and pharmaceutical residues for soil living organisms and plant (M4.1). With this aim two approaches will be applied. The first will rely on a literature review (rate-, duration- and frequency- of irrigation, type and amount of residues found,…) of exposure data under environmentally and agronomically relevant conditions and realistic exposure scenarios. The second will rely on the assessment of the fate of pesticide and pharmaceutical residues (including search for their main metabolites) in soil columns irrigated with wastewater to characterize their distribution between liquid and solid phases of soil. Having this data in hand, one could choose a range of model compounds (up to three compounds each) that could be applied solely at agronomically realistic concentrations to soil microcosms, incubated under control conditions, to assess their impact on soil-living organisms (earthworms and soil microorganisms) and plant (M4.2). These ecotoxicological studies will all include a negative control (amendment of soil with sterile water) and a positive control (amendment with 10 or 100 times the agronomical concentration). Impact on earthworms will be monitored by measuring the impact of chemical residues on a set of enzymatic activities as physiological traits of their health and functions (D4.4). Impact on soil microorganisms will be addressed by monitoring the abundance of different microbial groups by qPCR and the diversity by next generation sequencing of rRNA amplicons generated from soil DNA extracts. It will also be done by assessing the impact on biodegradation ability of soil microbial activity on a range of chemicals. Moreover, adaptation of soil microorganisms to pesticide and pharmaceutical residues will be searched by setting up repeated enrichment experiments and monitoring microbial degrading capabilities by radiorespirometric measurements (D4.2). Impact on plant will be assessed by monitoring production of plant hormones (auxin, cytokinins, jasmonates, abscisic acid) as a proxy of phyotoxicity caused by pesticides and pharmaceutical residues (D4.3). WP5. In WP5 the data from WPs 1 and 4 will be integrated (M5.3) and disseminated widely through the scientific community (papers, books chapters and communications in scientific meetings(D5.2, D5.3)), socio-economic sectors and regulatory bodies involved in water management and ultimately to the end-users.

D1.1 Report on efficiency of T. harzianum to remove pharmaceuticals 1 UM 12
D1.2 Report on enantiomeric fractionation tool 1 UM 18
D1.3 Report on attenuation of pesticide/pharmaceutical in soil 1 NIBIO/UM 36
D2.1 Vademecum of plant extraction methods 2 HMGU 12
D2.2 Kinetic data and stress parameters transferred to WP4 2 HMGU 24
D2.3 Report on metabolic pathways and products in plants 2 HMGU 30.
D2.4 Assessment of PPCP uptake, and evaluation of bioassays 2 CSIC 36
D3.1 Report on analytical results 3 UM 34
D3.2 Report on riperian strip zones to prevent pesticide runoff 3 NIBIO 35
D3.3 Report on risk assessment 3 UM 36
D4.1 Assessment of the impact of model pesticides and drugs on the diversity, abundance and activity of soil microorganisms 4 INRA 32
D4.2 Evaluation of the adaptation of soil microorganisms to biodegradation and repeated exposure to pesticide and drugs 4 INRA 34
D4.3 Estimation of the phytoxicity by monitoring the production of plant hormones in response pesticide and drugs residues 4 UM 34
D4.4 Assessment of L. terrestris role in PPCP degradation 4 CSIC 36
D4.5 Response of carboxylesterases to PPCP exposure 4 CSIC 36
D5.1 Recruitment advertisements 5 all 1
D5.2 Organization of a session in SETAC meeting (Rome 2018) 5 all 13
D5.3 Scientific book (manuscripts ready to be submitted) 5 CSIC 33
D5.4 Final meeting 5 CSIC 34

References coordinator and  leaders of  each WP:
WP1 - T. Eggen
WP2 - P. Schröder
WP3 - S. Chiron
WP4 - F. Martin-Laurent
WP5 - D. Barceló

Main outputs

  • Butkovskyi, A., Yuying, Y., Bergheim, H., Lazar, D., Gulyaeva, K., Odenmark, S.R., Norli, H.R., Nowak, K.M., Miltner, A., Kästner, M., Eggen, T. Retention and distribution of pesticides in planted filter microcosms designed for treatment of agricultural surface runoff. Science of the Total Environment (accepted 2021). (open access)
  • Solé, N. Montemurro, S. Pérez. Biomarker responses and metabolism in Lumbricus terrestris exposed to drugs of environmental concern, an in vivo and in vitro approach. Chemosphere.
  • Gallego, D. Nos, N. Montemurro, J.C. Sanchez Hernandez, S. Perez, M. Solè, F. Martin-Laurent. Ecotoxicological impact of the antihypertensive valsartan on earthworms, extracellular enzymes and soil bacterial communities. Environmental Pollution (2021) 275 – 116647.
  • Bigott, S. P. Chowdhury, S. Pérez, N. Montemurro, R. Manasfi, P. Schröder. Effect of the pharmaceuticals diclofenac and lamotrigine on stress responses and stress gene expression in lettuce (Lactuca sativa) at environmentally relevant concentrations. Journal of Hazardous Materials (2020).

More results on the project: Data and resources

Contact Point for  Communication/Dissemination activities: D. Barceló

Contact Point for Open Data/Open Access activities: D. Barceló

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published on 2017/03/23 11:00:00 GMT+2 last modified 2022-05-10T15:12:27+02:00