News

ENDETECH on the H2020 website!

Posted on February 9, 2015

The European Commission published an article about ENDETECH on the H2020 website. You can find it here: http://ec.europa.eu/programmes/horizon2020/en/news/enzymes-%E2%80%93-new-weapon-war-drugs-water

Two new papers published by CNRS-IEM

Posted on January 12, 2015

CNRS-IEM has recently published two papers on the research conducted in the context of the ENDETECH program.
Characterization of laccase-grafted ceramic membranes for pharmaceuticals degradation.
    M. de Cazes, M.-P. Belleville, M. Mougel, H. Kellner, J. Sanchez-Marcano.   
    J. Membrane Sci. 2015, 476, 384-393.
http://www.sciencedirect.com/science/article/pii/S0376738814008886
 
Large Scale Enzymatic Membrane Reactors for Tetracycline Degradation in WWWTP Effluents.
    R. Abejon, M. de Cazes, M.-P. Belleville, J. Sanchez-Marcano.   
    Water. Res. 2015, in press.

Happy New Year from ENDETECH team

Posted on January 5, 2015

The ENDETECH Team wishes you all a beautiful Holiday Season and a New Year of good health, happiness and prosperity!

GLOBAQUA-CYTOTHREAT-ENDETECH-SCARCE – Workshop Pharmaceuticals in Waters

Posted on December 1, 2014

The EU projects GLOBAQUA-CYTOTHREAT-ENDETECH funded under the European Union’s Seventh Framework Programm together with SCARCE from the Spanish Consolider Programm (Ministry of Economy and Competitiveness), have joined efforts to prepare a unique workshop in which pharmaceuticals will be considered from a multidisciplinary perspective. The workshop Pharmaceuticals in Wastewaters and Surface Waters under Multistressors Situation: Fate, Adverse effects, Risks and Removal Technologies will be held on 2-3 December 2014 in Barcelona, Spain.
You can find more information in the workshop flyer.
 

ENDETECH participates to the Adebiotech “Enzymes Innovation Industries” Workshop

Posted on September 25, 2014

The European Membrane Institute (IEM) will attend the Adebiotech Enzymes workshop scheduled from the 27th to the 28th of October in Paris, France to represent the ENDETECH program and share their recent findings on tetracycline-degradation in wastewaters with an enzymatic membrane reactor. A short presentation and a poster will be presented to the audience. The workshop program is available here.
 
 
Abstract of the presentation and poster (in French):
 
Poster #12 Session 6 - Conception et étude d’un bioréacteur enzymatiques à membrane pour le traitement d’effluents contenant des micropolluants réfractaires d’origine pharmaceutique
L’objectif de ce travail consiste à développer et optimiser un réacteur enzymatique à membrane permettant d’étudier l’élimination des polluants pharmaceutiques réfractaires présents dans les eaux usées. Pour ce faire, la tétracycline a été choisie comme molécule modèle afin d’étudier la transformation enzymatique des antibiotiques avec une membrane greffée avec la laccase de Trametes versicolor. Les résultats expérimentaux de dégradation de la tétracycline dans l’eau et à température ambiante (25°C et pH = 6) avec une quantité équivalente d’enzymes libres ou immobilisées, démontrent que les enzymes immobilisées sur un support membranaire en céramique présentent une meilleure réactivité et stabilité. L’étude des paramètres opératoires du procédé de dégradation a permis d’obtenir les conditions optimales de dégradation. Ce projet de recherche est développé dans le cadre du projet européen ENDETECH du programme Eco-innovation du FP7.

ENDETECH at the 13th Mediterranean Congress of Chemical Engineering (MCCE 2014)

Posted on September 5, 2014

The Mediterranean Congress of Chemical Engineering is a meeting which showcases a large number of the developments and contributions that chemistry has generated in order to increase and improve the health and welfare of humankind. The congress is a key event because it presented all the breakthroughs produced by Chemical Engineering in areas such as sustainable development in Environment, Food Technology and Biochemistry as well as innovations in traditional fields including Separation Technologies and Chemical Reactors. This congress will take place in Barcelona, Spain, from the 30th of September to the 3rd of October 2014. The European Membrane Institute will attend and present last results on Enzymatic membrane reactors for antibiotic degradation in wastewaters: tetracycline as case study.
 
Abstract of the presentation:
Antibiotics are not completely depleted by classical wastewater treatments, so conventional and MBR integrating wastewater treatment plants must incorporate specific advanced treatments for the elimination of this type of refractory micropollutants. Neither membrane technologies such as nanofiltration and reverse osmosis, nor adsorption on activated carbon (both options have been deeply taken into consideration for antibiotics removal) are able to transform these pharmaceuticals into less hazardous chemicals. By contrast, advanced oxidation processes under certain conditions can degrade antibiotics to biologically inactive compounds, hence they should be recommended as more effective treatment options. Oxidative enzymes have to be taken into account within the framework of advanced oxidation processes and they can be very useful for wastewater treatments and might be more interesting compared to other chemical or physical oxidative processes regarding toxicity of TPs generated after degradation of biologically active molecules, such as antibiotics.
Enzymatic membrane reactors are based on a semipermeable membrane which promotes the separation of the enzyme from the reaction mixture including substrates and/or products. Several enzymatic membrane reactor configurations have been tested, but more investigation is still required to advance in the design of this type of reactors and their direct application to real wastewater treatment processes for antibiotics degradation and other industrial processes (Ben Ameur et al., 2014).
The present work is related to the development of mathematical models in order to simulate the performance of enzymatic membrane reactors. They were applied to investigate the potentiality of laccase immobilized over ceramic membranes for the degradation of tetracycline, a very common antibiotic appearing as micropollutant in aquatic bodies, including wastewaters, even effluents from wastewater treatment plants (de Cazes et al., 2014). The technical viability of the proposed designs based on these simulated enzymatic membranes reactors connected in series has been demonstrated for the treatment of the different types of effluents (municipal, hospital and industrial pretreated wastewaters). For all the investigated cases, enzymatic membrane reactors appeared as realistic alternatives to other suggested treatments and they could be coupled to existing plants without suffering from severe space limitations due to disproportionate equipment dimensions (Abejón et al., 2014).

ENDETECH at the 4th International Conference on Industrial and Hazardous Waste Management

Posted on August 25, 2014

The International Conference on Industrial and Hazardous Waste Management will be held on September 2nd – 5th, 2014 in Chania (Crete, Greece) for the fourth time. The Conference will focus on innovative aspects of Industrial and Hazardous Waste Management (including Organics, Non-Halogenated and Halogenated Solvents, Hydrocarbons, Pesticides, Explosives, PCBs, PCDDs/Fs, Heavy Metals, Asbestos, Nuclear Wastes, etc), presenting new technologies, describing the state of the art and related case studies, discussing the main controversial subjects, sharing experience among different countries, valuating social and financial balances. The Catalan Institute for Water Research (ICRA) will attend and will present the last joint results of the ENDETECH consortium on Enzymatic decontamination processes for micropollutants: degradation studies and transformation products.
Abstract of the presentation:
The conventional treatment technologies for waters are very efficient on a large number of compounds. However, some of the chemical contaminants present in waters are not removed by these treatments since they are emitted to the media at higher amounts and are high stable (i.e. antibiotic residuals and endocrine disrupting chemicals among others). The chronic exposure to these compounds is a topic of threat as the long term effects are so far unpredictable and undocumented. For example, the presence of traces of antibiotics (and pharmaceuticals in general) in the environment can induce the development of antibiotic-resistant pathogens (Hirsch et al. 1999; Alighardashi et al. 2009; Li et al. 2010), causing problems in biota and for human health. The global objective of the ENDETECH EU-project is to develop a technology which aims at eliminating persistent pharmaceutical pollutants in wastewaters originating from drug manufacturing sites, households, hospitals and animal farms, thanks to an innovative ENzymatic DEcontamination TECHnology, by the use of different enzymes that alter the target chemicals by inactivating them (ENDETECH). The main work presented in these meeting deals with the degradation studies of the enzymatic decontamination processes developed within the frame of the project and the identification of transformation products under a chemical point of view.
Within the project, different enzymes have been tested for the inactivation of selected compounds, such as cyclines or macrolides, at laboratory scale and then extrapolated to a batch reactor system. During these studies, the presence of the selected compounds (i.e. tetracycline and erythromycin) has been assessed during the degradation process as well as the generation of any transformation product. In addition, the antibiotic activities for the tested compounds as well as the possible endocrine disruption have been evaluated along the biodegradation experiments.
The different degradation processes denoted the capabilities of the enzymatic decontamination as a complementary technology when the common removal process is not enough. For example, the degradation of tetracycline antibiotic by laccase enzyme from Trametes versicolor reached c.a. 78% after 18h of exposure without the presence of mediators at laboratory scale. When this process is transferred to a batch reactor, the degradation arrived to 60% (after 24h) for the enzymes immobilized in membranes while the degradation with free enzymes did not exceed the 30% . Another process optimized within the project has been the degradation of erythromycin by EreB esterase enzyme without the presence of mediators. In this case, the degradation of this compound reached the 50% after 16h of exposure. In these two examples the antibiotic activities were assessed along all the experiments decreasing c.a. 100%. As regards to the transformation products (TPs) detected during the experiments, different structures have been postulated after the analysis of the samples in a high resolution mass spectrometer Orbitrap. For example, during the degradation of tetracycline at laboratory scale three suspected TPs were proposed. The tentative molecular formulas of the molecular ions include C20H19N2O9 (TP 431), C20H14NO8 (TP 396) and C26H30N6O12 or C24H28N9O11 (TP 618), with mass errors within 2 ppm in most of the cases. The structures have been based on the hypothesis of dehydroxylation, (bi)demethylation and oxidation of the rings A and C as major reactions. In another example, the degradation of erythromycin leaded the tentative identification of five TPs. However, the major TP detected for this compound has been postulated as the “dehydration ERY-A” (TP 734), with the same molecular formula of its target compound and a mass error of 0.317 ppm, also identified in a previous study performed by Pengelly (2010).
 
The presentation of ICRA can be downloaded here.

ENDETECH participates to the “Pharmaceuticals in wastewaters and surface waters under multistressors situation: Fate, Adverse effects, Risks and Removal Technologies” Workshop

Posted on August 1, 2014

On the 2nd and 3rd of December 2014, the Globaqua-Cytothreat-Endetech-Scarce Workshop will be held in Barcelona, Spain. This event gathers the three EU projects funded under the European Union's Seventh Framework Program together with SCARCE from the Spanish Consolider Program.  It illustrates the shared willingness to organize a unique conference in which pharmaceutical pollutants will be considered from a multidisciplinary perspective. This is a unique opportunity to gather the European scientists and innovators involved in this research.
 
Upstream of this event, the 1st of December 2014, the ENDETECH project partners will meet for the last time in Barcelona, Spain.
 
This event is an opportunity for all the scientific committee of this project, and the others members present, to exchange concerning the development of a novel eco-efficient environmental technology whose use can substantially contribute to the reduction of polluting emissions.

ENDETECH at the 10th International Congress on Membranes and Membrane processes (ICOM2014)

Posted on July 21, 2014

The International Congress on Membranes and Membrane processes is known as one of the must-attend meeting in the membrane community. It will cover in 2014 all the aspects in membrane science and engineering, aiming to stimulate discussions on the cutting edge of research in membranes and membrane processes. Thus, the European Membrane Institute (IEM) will appear at this meeting scheduled from the 20th to the 25th of July in Suzhou, China, to represent the ENDETECH program and share their recent findings on tetracycline-degradation in wastewaters with an enzymatic membrane reactor. IEM will present results of laccases enzymes being grafted onto ceramic membrane within a 2L bioreactor with a continuing degradation of tetracycline during 100h from a solution at 20 mg/L of Tetracycline within osmosed water.

The presentation of IEM can be downloaded here.
 
Abstract of the presentation:
Traces of pharmaceutical pollutants are a threatening ecological issue, as pharmaceutical compounds are designed to be biologically active at very low concentrations in humans and animals. These compounds are relatively resistant to classical wastewater treatment techniques and their long term effects are so far unpredictable and undocumented. Among pharmaceuticals, antibiotics like tetracycline (TC) are frequently present in treated wastewaters and could become an important public health problem in a near future. Enzymatic membrane reactors (EMRs) can be considered as an original way to eliminate these recalcitrant pollutants in mild conditions [1].
The objective of this work is the degradation of TC from aqueous solutions in an EMR with laccase-grafted ceramic membranes. The active membranes were prepared according to a 3-step procedure which involves (i) the porous ceramic support coating with a gelatin solution, (ii) the cross-linking and activation with glutaraldehyde and finally (iii) the enzyme grafting [2]. The interest using porous ceramic supports is that they can be regenerated and reused for a further immobilization once the enzymes have lost their activity. The active membranes were prepared using as supports 0.2 µm and 1.4 µm monochanel membranes from Pall-Exekia and 0.2 µm 7 channels membranes from Tami Industries.
 Active membranes were firstly characterized by SEM. Results show that 0.2 µm membranes present a thin continuous gelatin-grafted enzyme layer at the surface, while for 1.4 µm membranes the layer was discontinuous and pores entrance is still visible. TC degradation experiments were carried out with 20 ppm tetracycline solutions in osmosed water. For this purpose, 15 cm length enzymatic membranes were placed in a pilot unit. The configuration used was in a batch mode by recirculating the retentate in the feeding tank. The runs were carried out in cycles by periodically replacing the TC solution with a fresh substrate solution and the TC concentration was analyzed continuously by HPLC.
An average tetracycline degradation rate of 124 mg.h-1.m-2 was reached with a 0.2 µm porous support, while this value was up to 275 mg.h-1.m-2 with 1.4 µm porous membrane. In this latter case it is possible that enzymes were not only immobilized on the surface but also within the pores, enabling a higher surface of contact with the substrate. It is important to note that no enzymatic activity drop could be observed after 4 cycles of reaction (for a total of 100 hours) proving that activity of enzymatic membranes is relatively stable.

ENDETECH at the 10th annual “LC/MS/MS Workshop on environmental applications and food safety”

Posted on June 28, 2014

At the 10th annual workshop on liquid chromatography-tandem mass spectrometry on environmental applications and food samples, the Catalan Institute for Water Research (ICRA) will represent ENDETECH program. ICRA will share their recent results on the identification of new transformation products of tetracycline and erythromycin antibiotics after enzymatic treatment.
This 10th annual workshop will take place in Barcelona, Spain, from the 1st to the 3rd of July, and aims to discuss the practical aspects of the use of tandem mass spectrometric techniques for screening and quantitation of organic contaminants in the environmental and food samples.
Using LC/MS/MS instruments, focused on environmental samples, ICRA has monitored the elimination of the antibiotic products as well as the generation of transformation products all along the water decontamination process by an enzymatic treatment technology developed in ENDETECH. This monitoring has been tested using a method based on the novel configuration of the on-line turbulent flow system coupled to the hybrid linear ion trap-high resolution mass spectrometer LTQ-Orbitrap.
 
Download the presentation of ICRA here.
 
Abstract of the poster presentation:
During last decades, antibiotic compounds have been considered as “emerging contaminants” due to their high consumption and pseudo-persistence in the environment. Waste water treatment plants (WWTPs) are one of the main sources of antibiotics into the environment, since these compounds are not completely eliminated during water treatment and are discharged through the wastewaters effluents into the river. Therefore, more efficient treatments for the elimination of these compounds are necessary.
This work describes the use of enzymatic decontamination process at laboratory scale for the degradation of two highly consumed antibiotics (tetracycline [Tc] and erythromycin [ERY], using laccase enzyme from Trametes versicolor and EreB esterase respectively). The analysis of the samples was carried out by a fast and simple method based on the novel configuration of the on-line turbulent flow system coupled to the hybrid linear ion trap-high resolution mass spectrometer LTQ -Orbitrap in order to monitor both (i) the elimination of the antibiotic along the treatment as well as (ii) the generation of any relevant transformation product (TP).
Degradation rates obtained for Tc and ERY were ~78% after 18 h and ~50% after 16 h, respectively. Concerning the identification of TPs after enzymatic treatment, the structures of three suspected compounds from Tc and five from ERY have been proposed. In the case of Tc, the tentative molecular formulas expressed as [M+H]+ of Tc TPs include C20H19N2O9 (TP 431), C20H14NO8 (TP 396) and C26H30N6O12 or C24H28N9O11 (TP 618), with errors mass within 2 ppm in most of the cases. The postulated structures have been based on the hypothesis of dehydroxylation, (bi)demethylation and  oxidation of the rings A and C as major degradation reactions of Tc. In contrast, the major TP detected for ERY has been identified as the “dehydration ERY-A” (TP 734), with the same molecular formula of ERY and an error mass of 0.317 ppm, which had been identified in a previous study (Pengelly 2010). In addition, the evaluation of the antibiotic activity of the samples along the enzymatic treatments showed 85% for Tc and qualitative decrease for ERY.
The promising results of enzymatic treatment (quantitative decrease of antibiotic concentration and antibiotic activity in the samples treated) allow us to recommend the enzymatic decontamination approach for the elimination of antibiotics.

ENDETECH at the 2nd international conference “EcoTechnologies for sewage treatments plants – ECOSTP2014”

Posted on June 3, 2014

The European Membrane Institute (IEM-CNRS) and the Catalan Institute for Water Research (ICRA) will represent the ENDETECH program on the 2nd IWA Specialized International Conference “EcoTechnologies for Wastewater Treatment (EcoSTP2014)" from the 23 to 25 June 2014. Launched in 2012 to provide a novel holistic approach to the “new conceived wastewater treatment plants”, this conference aims to gather experts, including engineers, scientists, economists and policy makers, to present their most recent technological and scientific results.
The European Membrane Institute will present its results to the ECOSTP2014 congress for the implementation of ceramic membranes grafted with laccase from Trametes versicolor in an enzymatic membrane reactor for the degradation of tetracycline. In addition, ICRA will present the identification of 5 transformation products for erythromycin and 3 for tetracycline after an enzymatic treatment by laccase from Trametes Versicolor and esterase EreB respectively.
 
Abstract of the IEM presentation:  The presence of pharmaceutical residues and endocrine disruptors in environmental water resources is a tough challenge as it is difficult to evaluate the potential impact of such a contamination on flora and fauna. If the low concentrations that are usually measured rule out the possibility of a direct toxic effect, few studies have measured the potential long-term hazard that emerges from the contamination at below-activity levels. With a limited efficiency of current wastewater treatment technologies to completely remove these contaminants, their treatment with an enzymatic membrane bioreactor appears as an interesting alternative. This work mainly describes the implementation of ceramic membranes grafted with laccase from Trametes versicolor in an enzymatic membrane reactor for the degradation of pharmaceuticals pollutants. The tetracycline was chosen as a model substrate to attest the grafting protocol optimization on two kinds of microfiltration membranes with pore diameters equal to 0.2 and 1.4 µm. The stability of different active membranes was tested to degrade a 20ppm tetracycline solution during several cycles of 24 hours.
Download here the presentation by IEM.
 
Abstract of the ICRA poster: During last decades, antibiotic compounds have been considered “emerging contaminants” due to their high consumption and pseudo-persistence in the environment being waste water treatment plants (WWTPs) one of the main inputs into the environment, since their elimination is far to be 100% for many of these compounds. This mild elimination represents an environmental problem since antibiotics can find their way to natural environment through the effluent wastewaters, since they are directly discharged into the river. This work describes the use of enzymatic decontamination process at laboratory scale for two highly consumed antibiotics (tetracycline (Tc) by laccase from Trametes versicolor and erythromycin (ERY) by EreB esterase) as a complementary treatment in WWTPs. The analysis of the samples was carried out by a fast and simple method based on the novel configuration of the on-line turbulent flow system coupled to the hybrid linear ion trap - high resolution mass spectrometer LTQ-Orbitrap in order to monitor the presence of any relevant transformation product (TP).
Download here the poster by ICRA.

ENDETECH to present at SETAC 2014

Posted on May 12, 2014

Among pharmaceutical pollutants considered for the ENDETECH project, carbamazepine is an antiepiletic drug unsufficiently removed during sewage treatment and particulary persistent in surface waters. The first results on carbamazepine ecotoxicity on sediment and benthic organisms will be presented by the Department for Aquatic Ecotoxicology from Goethe University Frankfurt to the SETAC Europe 24th Annual Meeting, which will be held from 11-15 May 2014 at Basel in Switzerland.

You can download the poster as presented by Dennis Becker at SETAC.

Abstract of the poster presentation:

Urban wastewaters contain among others a large variety of pharmaceutically active compounds and its metabolites. However, since the ubiquity of these compounds are only insufficiently degraded in wastewater treatment plants they are emitted into surface waters. After entering the aquatic environment freshwater species are exposed to an unknown, but probably very large number of compounds. Of particular concern is the long-term exposure of these aquatic organisms to low doses of pharmaceuticals. In order to elucidate these ecological impacts, Carbamazepine (CBZ) serves as a candidate for an environmentally persistent pharmaceutical. CBZ is an antiepileptic drug, which is only unsufficiently removed during sewage treatment.

Due to known effects of CBZ on Chironomus riparius larvae, these outcomes will be reinvestigated more precisely by looking at the sensitivity of different larval stages. Moreover the influence of CBZ on Culex pipiens f molestus will be examined as another representative within the suborder of midges.

A 28 day sediment test according to OECD guideline 218 was conducted with C. riparius. The experiment was carried out using four different larval stages exposed to spiked sediment. CBZ concentrations were 0.8, 2, 4, and 20 mg/kg dry weight sediment.

 C. pipiens f molestus was exposed to CBZ in a chronic 28 day experiment in the water phase to model their specific habitat. First instar larvae were exposed to the following CBZ concentrations 0.16, 0.32, 0.64, 1.28, 2.56, 5.12 and 10.24 mg/L. These concentrations refer to the measured concentrations in the water phase in Oetken et al.(2005).

First experiments indicate an impact of CBZ on C. riparius and C. pipiens f molestus.

Besides the indirect effects on pupation of C. riparius, a clear difference occurred regarding the sensitivity between first and fourth larval instars. The sensitive window of exposure for triggering the effect seems to be in the first or second larval stage. C. pipiens f molestus also showed a delay in emergence rate starting at a CBZ concentration of 2.56 mg/L. Additional tests with mayflies revealed a high sensitivity of the larvae towards CBZ.

Ongoing experiments will focus on the effects of CBZ metabolites and the influence of ozonation on the degradation of those compounds.

ENDETECH presents promising results on the design and optimization of an enzymatic membrane reactor for tetracycline degradation

Posted on April 20, 2014

The European Membrane Institute located in Montpellier, France, has published on April 2014 the first results of ENDETECH on the degradation of tetracycline in a pilot scale reactor. IEM has recently implemented a large-scale enzymatic membrane reactor with grafted laccase from T. versicolor for the continuous degradation of a Tetracycline solution.  Results have shown that grafted enzymes on membranes are more efficient for tetracycline degradation than free enzymes, and that active membranes can be used for several cycles of tetracycline degradation without enzymatic activity loss. Those results are thus promising regarding the objective of the ENDETECH program.
 
The article is available here: http://dx.doi.org/10.1016/j.cattod.2014.02.051
 
Abstract: Design and optimization of an enzymatic membrane reactor for tetracycline degradation
M. de Cazes, M.-P. Belleville, E. Petit, M. Llorca, S. Rodríguez-Mozaz, J. de Gunzburg, D. Barceló, J. Sanchez-Marcano
The tetracycline, antibiotic considered as a recalcitrant pollutant, was successfully depleted from model aqueous solutions by immobilized laccase from Trametes versicolor in an enzymatic membrane reactor. The results obtained show that tetracycline is depleted from water solutions at room temperature and without adding any extra chemicals. The degradation of tetracycline in aqueous solutions at 20 mg L−1 during 24 h, with equivalent amounts of free or immobilized biocatalyst, allowed reaching a tetracycline degradation yield of 56% with an enzymatic membrane whereas it was only of 30% with free laccase. This result highlights the good reactivity and stability of the immobilized enzyme for the degradation of tetracycline. Moreover, the enzymatic membrane reactor was able to reach a constant degradation rate of 0.34 mg of tetracycline per hour during 10 days.

ENDETECH at the 4th Green Process Engineering congress

Posted on April 12, 2014

Focused on technical innovations and green chemical processes, the 4th Green Process Engineering congress welcame the European Membrane Institute (IEM-CNRS) for a poster show from the 7th of April 2014 to the 10th April 2014 in Sevilla, Spain.

It was the opportunity for the team of IEM to present their last results on tetracycline-degradation experiments comparing the use of dense (α-alumina) or porous (gamma-alumina) ceramic beads grafted with laccase from Trametes Versicolor. Results demonstrated that the highest depletion yields were achieved with grafted porous gamma-alumina beads due to the coupling of the tetracycline adsorption and degradation phenomena.

Abstract of the poster:

The presence of pharmaceutical residues and endocrine disruptors in environmental water resources is a daunting challenge as it is difficult to evaluate the potential impact of such a contamination on flora and fauna. If the concentrations that are usually measured rule out the possibility of a direct toxic effect, few studies have measured the potential long-term hazard that emerges from the contamination at below-activity levels. With a limited efficiency of current wastewater treatment technologies to completely remove these contaminants, their treatment with an enzymatic membrane bioreactor appears as an interesting alternative. It represents an opportunity to avoid the pollution of water reservoirs such as groundwater that can hardly be depolluted in case of contamination by limiting the presence of pharmaceutical and endocrine-disrupting contaminants in effluents.

This work describes the comparison between dense (α-alumina) or porous (gamma-alumina) ceramic beads grafted with laccase from Trametes versicolor for tetracycline degradation. The tetracycline was chosen as a model substrate to attest the enzymatic degradation as far as this biocatalyst has already been used for the degradation of various pollutants present in water. The grafting protocol was optimized with dense beads to obtain the most efficient coating. The highest degradation yield was obtained with a 1 g.L-1 aminosilane solution and an enzymatic concentration of 10 g.L-1. The stability of the active support was also tested to see if it was reusable for further tetracycline degradation (4 cycles of 24 hours). The best depletion yields were achieved with grafted porous gamma-alumina beads due to the coupling of the adsorption-degradation phenomenon.
Degradation kinetics parameters were determined for free and immobilized enzymes on dense beads with two substrates: ABTS and tetracycline. Better results were obtained with grafted enzymes for both substrates because of a stability enhancement of the biocatalyst after immobilization. Scanning electron microscopy enabled to observe the structure of raw and active supports for both kinds of beads before and after reacting with the tetracycline.

You can download the poster as presented by Matthias de Cazes at the GPE congress by clicking here.

ENDETECH at the 24th ANALYTICA conference

Posted on March 1, 2014

The Catalan Institute for Water Research (ICRA) represented the ENDETECH program during the 24th ANALYTICA conference. The conference, which is certainly one of the most important conferences in the context of analytical sciences, was held in Munich, Germany on 1-3 April 2014.

As part of analytical sciences, water analysis and new emerging challenges were highlighted; among them, the issue of pharmaceuticals transformation products. D. Barceló from ICRA attended the congress for an oral presentation regarding the identification of new transformation products of selected pharmaceuticals caused by new-generation water-treatment processes, such as advanced oxidation, photolysis, and enzymatic treatment. Transformations products of antibiotics such as ofloxacine and tetracycline were identified by MS/MS experiments after (i) biological treatment using the fungi Trametes Versicolor and (ii) enzymatic treatment developed in the context of ENDETECH.

The presentation given by D.Barcelo is available here.

Abstract of the presentation:

Identification of new transformation products of selected pharmaceuticals formed during advanced oxidation, photolysis and enzymatic wastewater treatment processes by Orbitrap MS
Barceló, D., Perez, S., Rodriguez-Mozaz, S., Llorca, M., Gros M. &  Zonja, B.

Wastewater treatment plants effluents are considered one of the main entrance of pharmaceuticals into the aquatic environment. During wastewater treatment, pharmaceuticals are subject to different processes that can contribute to their partial elimination. Different transformations (biotic and abiotic) can occur during these processes, in occasions producing transformation products (TPs) of unknown toxicity and persistence. In the last years a great effort is being devoted to the identification of these unknown TPs. This type of study requires application of powerful analytical instruments in order to provide high-quality structural information needed for unequivocal identification of unknown compounds, which is of crucial importance in understanding the fate of organic contaminants in the environment. High mass resolution capability and mass accuracy provided by advanced mass spectrometric instruments such as Orbitrap Mass Spectrometer, enables the identification of possible transformation products. After determining the elemental composition based on accurate mass and isotopic pattern observed in the full scan MS spectrum MS/MS experiments are conducted in order to generate structural information. Although product ion spectra can also be generated using low resolution mass analyzers (e.g. triple quadrupole instruments (QqQ) and ion traps (IT)) fragment ions obtained under high resolution conditions afford far higher confidence in determining their chemical formula. Additionally, over the past several years, there has been an increase in the use of automated instruments that integrate extraction; purification and detection step in the analysis of organic emerging contaminants in environmental samples.

In this presentation, the results about the application of the Orbitrap technology for the study of new transformation products of pharmaceuticals will be discussed. Some recent studies of our group will be presented regarding the formation of the TPs of selected compounds after physico-chemical and biological processes applied in wastewater treatment. They include i) ii)   iii) identification of TPs of selected antibiotics, ofloxacine and tetracycline, after biological treatment using the fungi Trametes Versicolor and an enzymatic treatment respectively.  Finally, the capabilities of the on-line system Turboflow (TFC), for the clean-up of samples, coupled to an UHPLC-Orbitrap MS system will also be discussed.

Acknowledgements
This work has been supported by the Spanish Ministry of Economy and Competitiveness through the project DEGRAPHARMAC (CTQ2010-21776-C02), the European Union Project ENDETECH (FP7-ENV-2011-ECO-INNOVATION) and by the European Regional Development Fund (ERDF). This study has been co-financed by the European Union through the European Regional Development Fund (ERDF) and supported by the Generalitat de Catalunya (Consolidated Research Group: Water and Soil Quality Unit 2009-SGR-965).

Season's greetings from ENDETECH team

Posted on December 19, 2013

The ENDETECH Team wishes you all a beautiful Holiday Season and a New Year of good health, happiness and prosperity!

ENDETECH at ICWRER and Micropol & Ecohazard 2013

Posted on June 27, 2013

The Department for Aquatic Ecotoxicology from Goethe University Frankfurt represented ENDETECH in two symposia in June: the 6th International Conference on Water Resources and Environment Research (6th ICWRER) and the Micropol & Ecohazard 2013. The eco-toxicity assessment of various carrier materials were presented - this study took place in the ENDETECH program before starting the immobilization of enzymes on these carrier materials. Eluates were tested in various in vitro bioassays. Besides the carriers also first samples with pollutant and enzyme were assessed for potential toxic transformation products.

The 6th International Conference on Water Resources and Environment Research (6th ICWRER) was held in Koblenz, Germany, from the 3rd to the 7th of June 2013. The Department for Aquatic Ecotoxicology from Goethe University Frankfurt (D. Becker) has attended the congress for an oral presentation regarding the ecotoxicological evaluation of carrier materials and transformation products.

ENDETECH was also represented at the Micropol & Ecohazard 2013, the 8th IWA Specialist Conference on Assessment and Control of Micropollutants/Hazardous Substances in Water. This conference was held from the 16th to the 20th of June in Zurich, Switzerland. The Department for Aquatic Ecotoxicology from Goethe University Frankfurt (GU) (D. Becker, J. Oehlmann, M. Wagner) have attended the congress for a poster presentation regarding the ecotoxicological evaluation of carrier materials and transformation products.

Download

The oral presentation at the 6th ICWRER

The poster at the Micropol & Ecohazard 2013

Abstract

Background. Micropollutants comprise a plethora of synthetic or natural compounds (e.g., pharmaceuticals, hormones, and industrial chemicals) that are emitted into the environment. However, concerns have been raised regarding the vast amount of chemicals released, their specific toxicity (e.g., EDCs), and their limited degradation during conventional wastewater treatment (e.g., X-ray contrast media).  

The EU-project ENDETECH aims to develop a new technology to enzymatically degrade pharmaceutical pollutants. Priority targets considered for enzyme selection are antibiotics, EDCs and psychiatric drugs.

Aim. The aim of ecotoxicological research within ENDETECH is to evaluate the potential toxicity caused by transformation products generated during the enzymatic treatment. Instead of evaluating only the final technology, we take a proactive approach and accompany every step in the development of this technology.

Material & Methods. We use a battery of in vitro assays to investigate endocrine effects, dioxin-like effects, mutagenicity, and cytotoxicity. These assays will be applied to leachates from materials used in the process, target compounds, and enzymatically treated samples. As a start, eluates from different carrier materials used for enzyme immobilization have been tested, as well as a first sample of the degradation of one target compound (erythromycin) with the enzyme esterase EreB.

First results. The analysis of eluates from the different carrier materials revealed an endocrine effect resulting from migration from acrylic beads. Sand seems to be a promising alternative as carrier material compared to the beads. There were observed no toxic by-products and sand is lower in price. The evaluation of the treated and untreated samples with erythromycin and EreB showed no effect resulting from the generated transformation products.  

Conclusions. These first outcomes demonstrate how important it is to accompany the development of a new technology regarding the presence or formation of toxic leachates and transformation products. The early detection and solution of potential obstacles will decrease investment efforts and increase the acceptance of the new technology. It is cost-effective, eco-friendly and can be easily applied in existing wastewater treatment plants, in intensive animal farming, and in hospital or industrial effluents. It is designed to complement already existing technology and not to replace it.

ENDETECH annual meeting at Frankfurt - 13th and 14th of June 2013

Posted on June 25, 2013

The mid-term meeting of the ENDETECH project gathered together 5 of the 6 ENDETECH partners in Frankfurt: Da Volterra, Chiral Vision, European Membrane Institute, the Laboratory of aquatic ecotoxicology from Goethe University and the Catalan Institute for Water Research. Kindly organized by the Goethe University in their brand new Biologicum building, the meeting encouraged very rich discussions about the preliminary results and the future prospects to be considered before the conclusion of the project. The participation of the Scientific Advisory Board, composed of Pr. Paola Verlicchi of the University of Ferrera in Italy and Pr. Nick Voulvoulis of the Imperial College of London in the United Kingdom, during this mid-term meeting resulted in particularly successful discussions about the encouraging prospects of the ENDETECH project.

ENDETECH consortium members

ENDETECH at the 11th International Conference on Catalysis in Membrane Reactors

Posted on June 20, 2013

The ENDETECH project started experiments with antibiotics molecules. First outcomes obtained with tetracycline degradation by free and grafted enzymes were presented in an abstract submitted to the 11th International Conference on Catalysis in Membrane Reactors (11th ICCMR). This congress will be held in Porto, from the 7th to the 11th of July 2013.

The European Membrane Institute (IEM) and Catalan Institute for Water Research (ICRA) participated in this project by combining their results in one article to be published in a Special Issue of Catalysis Today. The IEM (M. de Cazes, M.-P. Belleville, J. Sanchez) will attend the congress for an oral communication regarding the “design and optimization of an enzymatic membrane reactor for tetracycline degradation”.

ICCMR11

Viral video from the Pharmas EU project

Posted on June 15, 2013

In order to demystify the issue of pharmaceutical pollutants in drinking water and in the environment, a short clip has been developed by the EU research project Pharmas, from the Pharma cluster.

ENDETECH at the International Congress on "Pharmaceutical Products in the Environment: Is there a problem?"

Posted on June 6, 2013

The ENDETECH project, based on the study of alternative decontamination processes based on enzymatic inactivation of antibiotics, presented the first results regarding tetracycline degradation by laccase enzymes and the transformation products generated during the treatment. The collaborative work developed by The European Membrane Institute (IEM) and the Catalan Institute for Water Research (ICRA) was presented in a poster communication during the congress that took place on 3-4th June 2013, Nîmes (France), under the title “Enzymatic degradation of tetracycline and generation of transformation products”.

The poster is available for download here.

The same work was presented in an oral communication during the “Pharmaceutical residues: parent compounds, metabolites and transformation products as environmental contaminants” Post Conference Workshop that took place on 5th June 2013, Nîmes (France).

Abstract

Tetracycline is a broad-spectrum polyketide antibiotic indicated for use against many human and veterinary bacterial infections such as urinary tract infections and acne among others. This antibiotic is consumed through oral, topical (skin and eye), intramuscular or intravenous application. Since this compound is hardly metabolized, tetracycline is eliminated from the body via fecal and renal excretions reaching waste water treatment plants (WWTP). Although the elimination/redistribution of this compound in WWTPs is between 60 and 70% (Gros et al. 2010), tetracycline is still present in effluents discharged into the river at low ng/L. The presence of traces of antibiotics (and pharmaceuticals in general) in the environment can induce the development of antibiotic-resistant pathogens, causing problems in biota and for human health (Hirsch et al. 1999; Bautitz et al. 2007). In this context, the evaluation of alternative treatments for its elimination should be considered such as enzymatic processes. This work is based in a novel enzymatic degradation process (de Gunzburg et al. 2012) - with 0.01 g/L of free Laccase enzyme from Trametes versicolor.

The main results outline elimination between 24 and 30% after 24 h of exposure. The possible transformation products generated during degradation process by enzymes have been studied by liquid chromatography coupled to a hybrid high resolution mass spectrometer LTQ-Orbitrap Velos (Thermo Fisher Scientific). Different sample pretreatment methods have been evaluated such as the novel on-line turbulent flow chromatography in order to remove potential matrix effects of the matrix as well as to recover not only tetracycline but the highest amount of their potential transformation products, which can cover a wide range of different physicochemical properties. The preliminary results indicate the generation of oxytetracycline, the (6S, 12aS)-3, 6, 10, 12, 12a-pentahydroxy-6-methyl-1, 4, 11-trioxo-1, 4, 4a, 5, 5a, 6, 11, 12a-octahydrotetracene-2-carboxamide and anhydrotetracycline.

The ENDETECH Consortium joins the Pharma Cluster

Posted on January 28, 2013

In the context of the collaboration between the European Union funded projects, ENDETECH is joining the innovation cluster PHARMA CLUSTER which is gathering the European projects dealing with the problematic of pharmaceutical products in the environment. All ENDETECH partners hope that the participation to the PHARMA CLUSTER will offer a strong scientific stimulation and will be an opportunity to improve significantly the scientific expertise in this area of great importance for our territories’ future.

The cluster is based on:

  • Exchange of research data and research plans in areas of overlapping interest areas through visits by scientific and/or technical staff of partner institutes;
  • Common dissemination strategy through cluster website (linked to individual project websites), newsletters, brochures;
  • Coordinated cluster press releases.

 

Season's Greetings from ENDETECH Team

Posted on December 28, 2012

The ENDETECH Team wishes you all a beautiful Holiday Season and a New Year of good health, happiness and prosperity!

Novel website for ENDETECH

Posted on July 19, 2012

The ENDETECH partners are pleased to announce the release of the project public website. This website will allow you to follow the activities and achievements of the consortium and contact the collaborative program partners or coordinator for further information. It will be regularly updated with information about the progress made by the project to reduce the ecological footprint of recalcitrant pharmaceutical pollutants.

Keeping in touch with social media, the ENDETECH consortium is also embracing Google Plus, Twitter and Facebook, with pages dedicated to the project on the three social platforms. Feel free to follow us!

ENDETECH Project Launch in Paris

Posted on February 8, 2012

The kick-off meeting of the ENDETECH program has launched the 3-year project in a warm and dynamic spirit. Gathered around Da Volterra, the coordinator of the ENDETECH programme, the 5 partners originating from 4 different European countries, braved the Parisian winter to share their vision of the project, their passion and ambition for its success.

c-LEcta and the Goethe Frankfurt University (Germany), ChiralVision (The Netherlands), Da Volterra and the European Membrane Institute (France) and the Catalan Institute for Water Research (Spain) met in the ‘Maison des Polytechniciens’ in the heart of Paris to present their skills and their expertise in the field of water treatment technologies and start the work on the ENDETECH collaborative program.

All partners present at the kick-off meeting took part to very rich and promising discussions related to the future of the project. They started working on the first ENDETECH tasks once back home - they are expected to present their first results in the coming months.