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Distribution of pesticides and some of their transformation products in a small lentic waterbody: Fish, water, and sediment contamination in an agricultural watershed
2021
Slaby, Sylvain | Le Cor, François | Dufour, Vincent | Auger, Lucile | Pasquini, Laure | Cardoso, Olivier | Curtet, Laurence | Baudoin, Jean-Marc | Wiest, Laure | Vulliet, Emmanuelle | Feidt, Cyril | Dauchy, Xavier | BANAS, Damien
Pregnancy exposure to synthetic phenols and placental DNA methylation -An epigenome-wide association study in male infants from the EDEN cohort ☆
2021
Jedynak, Paulina | Tost, Jörg | Calafat, Antonia, M | Bourova-Flin, Ekaterina | Busato, Florence | Forhan, Anne | Heude, Barbara | Jakobi, Milan | Rousseaux, Sophie | Schwartz, Joel | Slama, Rémy | Vaiman, Daniel | Philippat, Claire | Lepeule, Johanna | Université Grenoble Alpes (UGA) | Centre for Research in Epidemiology and Statistics | Centre de Recherche Épidémiologie et Statistiques (CRESS (U1153 / UMR_A 1125)) ; Conservatoire National des Arts et Métiers [CNAM] (CNAM) ; HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Université Sorbonne Paris Cité (USPC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
International audience | In utero exposure to environmental chemicals, such as synthetic phenols, may alter DNA methylation in different tissues, including placenta - a critical organ for fetal development. We studied associations between prenatal urinary biomarker concentrations of synthetic phenols and placental DNA methylation. Our study involved 202 mother-son pairs from the French EDEN cohort. Nine phenols were measured in spot urine samples collected between 22 and 29 gestational weeks. We performed DNA methylation analysis of the fetal side of placental tissues using the IlluminaHM450 BeadChips. We evaluated methylation changes of individual CpGs in an adjusted epigenome-wide association study (EWAS) and identified differentially methylated regions (DMRs). We performed mediation analysis to test whether placental tissue heterogeneity mediated the association between urinary phenol concentrations and DNA methylation. We identified 46 significant DMRs (>= 5 CpGs) associated with triclosan (37 DMRs), 2,4-dichlorophenol (3), benzophenone-3 (3), methyl- (2) and propylparaben (1). All but 2 DMRs were positively associated with phenol concentrations. Out of the 46 identified DMRs, 7 (6 for triclosan) encompassed imprinted genes (APC, FOXG1, GNAS, GNASAS, MIR886, PEG10, SGCE), which represented a significant enrichment. Other identified DMRs encompassed genes encoding proteins responsible for cell signaling, transmembrane transport, cell adhesion, inflammatory, apoptotic and immunological response, genes encoding transcription factors, histones, tumor suppressors, genes involved in tumorigenesis and several cancer risk biomarkers. Mediation analysis suggested that placental cell heterogeneity may partly explain these associations. This is the first study describing the genome-wide modifications of placental DNA methylation associated with pregnancy exposure to synthetic phenols or their precursors. Our results suggest that cell heterogeneity might mediate the effects of triclosan exposure on placental DNA methylation. Additionally, the enrichment of imprinted genes within the DMRs suggests mechanisms by which certain exposures, mainly to triclosan, could affect fetal development.
Afficher plus [+] Moins [-]Pregnancy exposure to synthetic phenols and placental DNA methylation -An epigenome-wide association study in male infants from the EDEN cohort ☆
2021
Jedynak, Paulina | Tost, Jörg | Calafat, Antonia, M | Bourova-Flin, Ekaterina | Busato, Florence | Forhan, Anne | Heude, Barbara | Jakobi, Milan | Rousseaux, Sophie | Schwartz, Joel | Slama, Rémy | Vaiman, Daniel | Philippat, Claire | Lepeule, Johanna | Université Grenoble Alpes (UGA) | Centre for Research in Epidemiology and Statistics | Centre de Recherche Épidémiologie et Statistiques (CRESS (U1153 / UMR_A 1125)) ; Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Université Sorbonne Paris Cité (USPC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
International audience | In utero exposure to environmental chemicals, such as synthetic phenols, may alter DNA methylation in different tissues, including placenta - a critical organ for fetal development. We studied associations between prenatal urinary biomarker concentrations of synthetic phenols and placental DNA methylation. Our study involved 202 mother-son pairs from the French EDEN cohort. Nine phenols were measured in spot urine samples collected between 22 and 29 gestational weeks. We performed DNA methylation analysis of the fetal side of placental tissues using the IlluminaHM450 BeadChips. We evaluated methylation changes of individual CpGs in an adjusted epigenome-wide association study (EWAS) and identified differentially methylated regions (DMRs). We performed mediation analysis to test whether placental tissue heterogeneity mediated the association between urinary phenol concentrations and DNA methylation. We identified 46 significant DMRs (>= 5 CpGs) associated with triclosan (37 DMRs), 2,4-dichlorophenol (3), benzophenone-3 (3), methyl- (2) and propylparaben (1). All but 2 DMRs were positively associated with phenol concentrations. Out of the 46 identified DMRs, 7 (6 for triclosan) encompassed imprinted genes (APC, FOXG1, GNAS, GNASAS, MIR886, PEG10, SGCE), which represented a significant enrichment. Other identified DMRs encompassed genes encoding proteins responsible for cell signaling, transmembrane transport, cell adhesion, inflammatory, apoptotic and immunological response, genes encoding transcription factors, histones, tumor suppressors, genes involved in tumorigenesis and several cancer risk biomarkers. Mediation analysis suggested that placental cell heterogeneity may partly explain these associations. This is the first study describing the genome-wide modifications of placental DNA methylation associated with pregnancy exposure to synthetic phenols or their precursors. Our results suggest that cell heterogeneity might mediate the effects of triclosan exposure on placental DNA methylation. Additionally, the enrichment of imprinted genes within the DMRs suggests mechanisms by which certain exposures, mainly to triclosan, could affect fetal development.
Afficher plus [+] Moins [-]Fate of nanoplastics in the environment: Implication of the cigarette butts
2021
El Hadri, Hind | Lisa, Jesus Maza | Gigault, Julien | Reynaud, Stephanie | Grassl, Bruno | Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux (IPREM) ; Université de Pau et des Pays de l'Adour (UPPA)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Centre National de la Recherche Scientifique (CNRS) | Géosciences Rennes (GR) ; Université de Rennes (UR)-Institut national des sciences de l'Univers (INSU - CNRS)-Observatoire des Sciences de l'Univers de Rennes (OSUR) ; Université de Rennes (UR)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Rennes 2 (UR2)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Rennes 2 (UR2)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Centre National de la Recherche Scientifique (CNRS) | Agence Nationale de la Recherche | ANR-16-IDEX-0002,E2S,E2S(2016)
International audience | Fate, transport and accumulation of nanoplastics have attracted considerable attention in the past few years. While actual researches have been focused on nanoplastics dispersed or aggregated in different environmental system, no study have been focused on the possibility that nanoplastics are co-transported with other natural or anthropogenic materials. Therefore, the large quantity of debris released in the environment, such as cigarette butts (CGB), could be part of the nanoplastics fate and behavior. Here we show the considerable sorption capacities of cigarette filters for nanoplastics. To address this topic, we chose polystyrene-based nanoplastics with similar state of charge (according to the physico-chemical characteristic of the zeta potential −45 to −40 mV) but with different sizes (50–800 nm) and morphologies. A kinetic approach to sorption in fresh water (pH = 8.05; 179.5 μS cm−1) at room temperature was carried out by means of the flow field flow analysis method (AF4) to determine the partition coefficients and water sampling rates between nanoplastics and cigarette butts. Using different models of, more or less environmentally relevant, nanoplastics (NPTs) and adequate analytical strategies, we found partition coefficients between the NPTs and CGBs ranged from 102 to 104 in freshwater conditions. We demonstrated that the physical features of the NPTs (size and morphology) have an influence on the sorption behaviour. Asymmetrical shaped NPTs with broader size distribution seems to be mostly retained in the CGBs after longer equilibration time. This result shows the importance of the NPTs features on the mechanisms governing their transfer and fate in the environment through environmental matrices, especially when other materials are involved. We anticipate our work to be a starting point for investigating the co-transport of NPTs with other materials present in the environment (natural and anthropogenic).
Afficher plus [+] Moins [-]Ecotoxicological impact of the antihypertensive valsartan on earthworms, extracellular enzymes and soil bacterial communities
2021
Gallego, Sara | Nos, David | Montemurro, Nicola | Sanchez-Hernandez, Juan | Pérez, Sandra | Solé, Montserrat | Martin-Laurent, Fabrice | Agroécologie [Dijon] ; Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Institute of Environmental Assessment and Water Research (IDAEA) ; Consejo Superior de Investigaciones Cientificas = Spanish National Research Council (CSIC) | Inst Marine Sci ICM CSIC, Renewable Marine Resources Dept, Barcelona, Spain. | Univ Castilla La Mancha, Inst Environm Sci ICAM, Lab Ecotoxicol, Toledo 45071, Spain
International audience | The use of reclaimed water in agriculture represents a promising alternative to relieve pressure on freshwater supplies, especially in arid or semiarid regions facing water scarcity. However, this implies introducing micropollutants such as pharmaceutical residues into the environment. The fate and the ecotoxicological impact of valsartan, an antihypertensive drug frequently detected in wastewater effluents, were evaluated in soil-earthworm microcosms. Valsartan dissipation in the soil was concomitant with valsartan acid formation. Although both valsartan and valsartan acid accumulated in earthworms, no effect was observed on biomarkers of exposure (acetylcholinesterase, glutathione S-transferase and carboxylesterase activities). The geometric mean index of soil enzyme activity increased in the soils containing earthworms, regardless of the presence of valsartan. Therefore, earthworms increased soil carboxylesterase, dehydrogenase, alkaline phosphatase, beta-glucosidase, urease and protease activities. Although bacterial richness significantly decreased following valsartan exposure, this trend was enhanced in the presence of earthworms with a significant impact on both alpha and beta microbial diversity. The operational taxonomic units involved in these changes were related to four (Proteobacteria, Bacteroidetes, Actinobacteria and Firmicutes) of the eight most abundant phyla. Their relative abundances significantly increased in the valsartan-treated soils containing earthworms, suggesting the presence of potential valsartan degraders. The ecotoxicological effect of valsartan on microbes was strongly altered in the earthworm-added soils, hence the importance of considering synergistic effects of different soil organisms in the environmental risk assessment of pharmaceutical active compounds. (C) 2021 Elsevier Ltd. All rights reserved.
Afficher plus [+] Moins [-]LONG-TERM stability of arsenic in iron amended contaminated soil
2021
Kumpiene, Jurate | Carabante, Ivan | Kasiuliene, Alfreda | Austruy, Annabelle | Mench, Michel | Luleå University of Technology = Luleå Tekniska Universitet (LUT) | Institut Ecocitoyen pour la Connaissance des Pollutions [Fos-sur-Mer] | Biodiversité, Gènes & Communautés (BioGeCo) ; Université de Bordeaux (UB)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | The La Combe de Saut experiment was funded by ADEME and the LIFE project DIFPOLMINE. The Reppel experiment was set up under the EU FP5 Phytorehab project. Experiments in Sweden were financed by the Swedish Research Council FORMAS and European Union Structural Funds and New Objective 1, North Sweden Soil Remediation Center (MarksaneringsCentrum Norr - MCN). Stanford Synchrotron Radiation Lightsource is acknowledged for granting our beamtime proposal (4663) under which X-Ray absorption measurements presented in this work were performed.
International audience | This study aimed at elucidating the long-term efficiency of soil remediation where chemical stabilization of arsenic (As) contaminated soil using zerovalent iron (Fe) amendments was applied. A combination of chemical extraction and extended X-Ray absorption fine structure (EXAFS) spectroscopy technique was applied on soils collected from five laboratory and field experiments in Sweden and France. All soils were treated with 1 wt% of zerovalent Fe grit 2e15 years prior to the sampling. The results indicate that all studied soils, despite the elapsed time since their amendment with Fe grit, had substantial amounts of ferrihydrite and/or lepidocrocite. These metastable and the most reactive Fe (oxyhydr)oxides (mainly ferrihydrite) were still present in substantial amounts even in the soil that was treated 15 years prior to the sampling and contributed most to the As immobilisation in the amended soils. This increases confidence in the long-term efficiency of As immobilisation using zerovalent Fe amendments. Both applied methods, sequential extraction and EXAFS, were in line for most of the samples in terms of their ability to highlight As immobilisation by poorly crystalline Fe phases.
Afficher plus [+] Moins [-]Changes in spatial patterns of ammonia dry deposition flux and deposition threshold exceedance according to dispersion model formalism and horizontal resolution
2021
Azouz, Niramson | Beekmann, Matthias | Siour, Guillaume | Cellier, Pierre | Drouet, Jean-Louis | Ecologie fonctionnelle et écotoxicologie des agroécosystèmes (ECOSYS) ; AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Laboratoire Interuniversitaire des Systèmes Atmosphériques (LISA (UMR_7583)) ; Institut national des sciences de l'Univers (INSU - CNRS)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité) | Institut Pierre-Simon-Laplace (IPSL (FR_636)) ; École normale supérieure - Paris (ENS-PSL) ; Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut national des sciences de l'Univers (INSU - CNRS)-École polytechnique (X) ; Institut Polytechnique de Paris (IP Paris)-Institut Polytechnique de Paris (IP Paris)-Centre National d'Études Spatiales [Toulouse] (CNES)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité) | ANR-12-AGRO-0003,ESCAPADE,Evaluation de Scénarios sur la Cascade de l'Azote dans les Paysages Agricoles et moDElisation territoriale(2012) | European Project: 282910,EC:FP7:ENV,FP7-ENV-2011,ECLAIRE(2011)
International audience | Ammonia (NH 3) emitted into the atmosphere from agricultural sources may affect nearby sensitive ecosystems due to high dry deposition fluxes on vegetation and soil surfaces, contributing to critical load exceedances. Ammonia fluxes near sources are simulated by either short-range atmospheric models or regional models using large grid cell sizes. However, studies are missing on the comparison of the results simulated by these two types of models. This paper presents the effect of model formalism, input factors, especially grid cell size and wind speed and the choice of deposition threshold on the spatial patterns of NH 3 dry deposition fluxes and deposition threshold exceedances. We used the Eulerian chemistry-transport model CHIMERE and the Gaussian plume model OPS-ST on two study domains characterised by contrasting land use. We showed that the average annual NH 3 dry deposition fluxes over each whole domain are similar for both models. By contrast, NH 3 dry deposition fluxes near sources are higher when simulated with OPS-ST that provides analytical solutions that can be sampled with small grid cell sizes (i.e., from 25 to 1600 m in this study), than with CHIMERE, which uses large grid cell sizes (i.e., 800 and 1600 m). As a result, the spatial patterns of deposition threshold exceedance were very different between both models. These patterns depend mainly on grid cell size, the input factors and the choice of the deposition threshold value. We show that the model formalism has a relatively small effect on the results and that the differences result mainly from the spatial resolutions to which they can be applied. Simulation results must therefore be interpreted carefully, taking into account the simulation conditions.
Afficher plus [+] Moins [-]Experimental and DFT investigation on N-functionalized biochars for enhanced removal of Cr(VI)
2021
Zhao, Nan | Zhao, Chuanfang | Liu, Kunyuan | Zhang, Weihua | Tsang, Daniel C.W. | Yang, Zaikuan | Yang, Xixiang | Yan, Bofang | Morel, Jean Louis | Qiu, Rongliang
International audience | In this study, N-functionalized biochars with varied structural characteristics were designed by loading poplar leaf with different amounts of urea at 1:1 and 1:3 ratios through pyrolysis method. The addition of urea significantly increased the N content of biochar and facilitated the formation of amine (-NH-, -NH2), imine (-HC=NH), benzimidazole (-C7H5N2), imidazole (-C3H3N2), and pyrimidine (-C4H3N2) groups due to substitution reaction and Maillard reaction. The effect of pH on Cr(VI) removal suggested that decrease in solution pH favored the formation of electrostatic attraction between the protonated functional groups and HCrO4-. And, experimental and density functional theory study were used to probe adsorption behaviors and adsorption mechanism which N-functionalized biochars interacted with Cr(VI). The protonation energy calculations indicated that N atoms in newly formed N-containing groups were better proton acceptors. Adsorption kinetics and isotherm experiments exhibited that N-functionalized biochars had greater removal rate and removal capacity for Cr(VI). The removal rate of Cr(VI) on N-functionalized biochar was 10.5-15.5 times that of untreated biochar. Meanwhile, N-functionalized biochar of NB3 with the largest number of adsorption sites for-C7H5N2,-NH2,-OH,-C3H3N2, and phthalic acid (-C8H5O4) exhibited the supreme adsorption capacity for Cr(VI) through H bonds and the highest adsorption energy was -5.01 kcal/mol. These mechanistic findings on the protonation and adsorption capacity are useful for better understanding the functions of N-functionalized biochars, thereby providing a guide for their use in various environmental applications.
Afficher plus [+] Moins [-]The combination of chemical, structural, and functional indicators to evaluate the anthropogenic impacts on agricultural stream ecosystems
2021
Jabiol, Jérémy | Chauvet, Eric | Guérold, François | Bouquerel, Jonathan | Usseglio-Polatera, Philippe | Artigas, Joan | Margoum, Christelle | Le Dréau, Matthieu | Moreira, Aurélie | Mazzella, Nicolas | Gouy, Véronique | Hydrobiologie et Fonctionnement des Ecosystèmes (HYFE) | Laboratoire Ecologie Fonctionnelle et Environnement (LEFE) ; Institut Ecologie et Environnement - CNRS Ecologie et Environnement (INEE-CNRS) ; Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3) ; Université de Toulouse (UT)-Université de Toulouse (UT)-Observatoire Midi-Pyrénées (OMP) ; Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3) ; Université de Toulouse (UT)-Université de Toulouse (UT)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP) ; Université de Toulouse (UT) | Laboratoire Interdisciplinaire des Environnements Continentaux (LIEC) ; Institut Ecologie et Environnement - CNRS Ecologie et Environnement (INEE-CNRS) ; Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Observatoire Terre et Environnement de Lorraine (OTELo) ; Institut national des sciences de l'Univers (INSU - CNRS)-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS) | Laboratoire Microorganismes : Génome et Environnement (LMGE) ; Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne (UCA) | RiverLy - Fonctionnement des hydrosystèmes (RiverLy) ; Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Ecosystèmes aquatiques et changements globaux (UR EABX) ; Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
International audience
Afficher plus [+] Moins [-]Effect of Filter Media and Depth on Hydrodynamics and Treatment Performances of Single-Stage French Vertical Flow Treatment Wetlands Treating Domestic Effluent
2021
Ruiz-Ocampo, Hernán | Tondera, K. | Katusic, Vlatka | Paing, Joëlle | Molle, Pascal | Chazarenc, Florent | Laboratoire de génie des procédés - environnement - agroalimentaire (GEPEA) ; Institut Universitaire de Technologie - Nantes (IUT Nantes) ; Université de Nantes (UN)-Université de Nantes (UN)-Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST) ; Université de Nantes (UN)-Université de Nantes (UN)-Institut Universitaire de Technologie Saint-Nazaire (IUT Saint-Nazaire) ; Université de Nantes (UN)-Ecole Polytechnique de l'Université de Nantes (EPUN) ; Université de Nantes (UN)-École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Centre National de la Recherche Scientifique (CNRS)-Université Bretagne Loire (UBL)-IMT Atlantique (IMT Atlantique) ; Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-Institut Universitaire de Technologie - La Roche-sur-Yon (IUT La Roche-sur-Yon) ; Université de Nantes (UN) | École des Ponts ParisTech (ENPC) | Réduire, valoriser, réutiliser les ressources des eaux résiduaires (UR REVERSAAL) ; Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | CREA step | Center-Val de Loire Region | French National Research Agency (ANR)
International audience | Treatment wetlands for raw wastewater from small communities (< 2000 p.e.) represent 20% of the wastewater treatment plants in France today. The classical French Vertical Flow Treatment Wetland consists of two stages filter beds intermittently fed with raw wastewater (inflow concentrations TSS 387 +/- 70 mg/L, COD 880 +/- 188 mg/L, TKN 110 +/- 16 mg/L). The system has a good removal performance for TSS and COD with > 90% and TKN of approx. 85% after filter maturation (months 12 to 24 of the trial), but efficiencies can vary between 10 and approx. 20% according to the material type, filter age, and filter depth. This study presents how these systems can be costly optimized when outlet requirements are not too stringent or when carbon is required for a subsequent denitrification step. The study shows the effectiveness of using different gravel depths and types (pea and crushed gravel) over almost 2 years regarding removal performances, carbon source availability, and nitrification. Core samples were taken to evaluate the dry matter accumulation. Tracer tests using fluorescein were performed to assess the internal hydrodynamics related to filter depth. The removal performance was not significantly affected by the filter depth, but the filter filled with 30 cm of pea gravel performed significantly better than the one with 30 cm of crushed gravel: for COD with a removal of 83% compared to 76%, TSS of 87% to 81%, and TKN of 64% to 57%, respectively. This is another indication that the shape of crushed gravel affects biofilm attachment and hydrodynamics in the filters due to irregular compaction.
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