Project presentation

damia barcelo culleres 

Damià Barceló Culleres

Executive Coordinator 

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

Two main issues have a high impact in the agricultural food production: the lack of water resources in some European countries and pollution with pesticides. The use of pesticides for pest control is a common practice. However, the usage patterns of water among European countries differ, e.g. Scandinavian countries 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. By means of specific regional water management practices, there is an introduction of different organic pollutants to the soil crops. Accordingly, the AWARE project aims to investigate the fate and potential reduction of pesticides and wastewater-borne contaminants in soil/plants from agricultural crops. Moreover, in the AWARE project we will evaluate environmental risks in agricultural fields due to the use of pesticides and the irrigation practices involving reused wastewater. Both pollution sources may have some inherent risks associated to food production. These goals will be addressed by: i) assessing the microbial degradation of pharmaceuticals in the rhizosphere-dynamic of antibiotic resistance genes as well as by studying the kinetics, metabolism, and fate including formation/sorption of non-extractable residues (NER) (OECD 307, 308) of selected pesticides (NIBIO, UFZ, UM), ii) examining uptake and metabolism of selected contaminants in batchreactor-grown plants under two scenarios using either spiked wastewater (worst-case scenario) or real wastewater (CSIC, HMGU); iii) estimating the ecotoxicological impact of targeted contaminants on soil microbial diversity and functions (INRA); iv) evaluating the effects of wastewater reuse in soil and plants on soil-dwelling earthworms, for their involvement in chemical degradation and for the role of earthworm (carboxyl)esterases as biomarkers of pharmaceutical and pesticide exposures (CSIC), v) investigating the effect of riparian zone and advanced designed wetlands to prevent runoff of high-mobile pesticides (NIBIO, UFZ), and vi) assessing at real scale the risks of using wastewater for irrigating crops (UM, HMGU). These challenges and objectives will be addressed through a multidisciplinary pan-european team including experts in environmental chemistry, analytical chemistry, plant physiology, toxicology, risk assessment, remediation and environmental technology processes. Therefore, the added-value of the partnership is related to the different expertise involved and the perfect combination between Universities and Research Institutions. Especially, the AWARE team members have a track record of working with local stakeholders informing them about novel developments. Finally, 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:  


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

Contact Point for  Communication/Dissemination activities:

D. Barceló

Contact Point for Open Data/Open Access activities:

D. Barceló

Picture of the research team:

 S. Pérez, A. Ginebreda, P. Schröder , C. Huber, T. Eggen, S. Chiron, E. Gomez, M. Kästner,

J. Kværner, G.Tveit, C.Coutris, S.R., Odensmarck, M.Almdal, F. Martin-Laurent, M. Sole, S.Chiron, H. Fenet