Alyssa Azaroff

Mathilde Monperrus

Carole Miossec

Claire Gassie

Rémy Guyoneaud

Microbial degradation of hydrophobic emerging contaminants from marine sediment slurries (Capbreton Canyon) to pure bacterial strain

published or not. The documents may come

Microbial degradation of hydrophobic emerging contaminants from marine sediment slurries (Capbreton

Canyon) to pure bacterial strain

Introduction

As a result of the last century, Man created and is still creating new synthetic substances. Although well known persistent organic contaminants, such as organochlorine pesticides or polycyclic aromatic hydrocarbons, are regulated in industrialized countries (European Commission, US, Japan) for several decades, regulation for emerging contaminants (ECs) arised only in the early of the 21th century and ⁎ Corresponding author.

E-mail address: remy.guyoneaud@univ-pau.fr (R. Guyoneaud).

T their update is still ongoing in Europe (Directive 2008/105/EC and Commission implementing decision 2018/840). Indeed, the scarce information available of their occurrence, reactivity and impact have led to a rising interest in identifying and screening these new compounds in the environment [START_REF] Field | What is "emerging[END_REF]. Among those ECs, pharmaceuticals and personal care products (PPCPs) are substances widely consumed and continuously released in the environment, mainly through wastewater, both treated and untreated [START_REF] Li | Occurrence, sources, and fate of pharmaceuticals in aquatic environment and soil[END_REF][START_REF] Rainieri | Occurrence and toxicity of musks and UV filters in the marine environment[END_REF].

Marine ecosystem is the final receptor for these organic ECs. They were found in marine sediments, mainly close to the high density population coast nearby the main estuaries [START_REF] Beretta | Occurrence of pharmaceutical and personal care products (PPCPs) in marine sediments in the Todos os Santos Bay and the north coast of Salvador, Bahia, brazil[END_REF][START_REF] Huang | Occurrence and distribution of synthetic musks and organic UV filters from riverine and coastal sediments in the Pearl River estuary of China[END_REF][START_REF] Pintado-Herrera | Distribution, mass inventories, and ecological risk assessment of legacy and emerging contaminants in sediments from the Pearl River Estuary in China[END_REF][START_REF] Azaroff | Priority and emerging micropollutants distribution from coastal to continental slope sediments: a case study of Capbreton Submarine Canyon (North Atlantic Ocean)[END_REF].

Marine sediments act as integrative matrices reflecting the pollution state in a given area [START_REF] Bellas | Integrative assessment of coastal pollution: development and evaluation of sediment quality criteria from chemical contamination and ecotoxicological data[END_REF][START_REF] Roberts | Causes and ecological effects of resuspended contaminated sediments (RCS) in marine environments[END_REF]. The affinities of contaminants with the suspended particulate matter (SPM) lead them to be readily scavenged from the water column and to be deposited in the sediments. Submarine canyons are known to act as transfer zones of suspended particulate matter and contaminants between the continent and the open ocean where organic pollutants can be accumulated [START_REF] Azaroff | Priority and emerging micropollutants distribution from coastal to continental slope sediments: a case study of Capbreton Submarine Canyon (North Atlantic Ocean)[END_REF][START_REF] Paull | Pesticides as tracers of sediment transport through Monterey Canyon[END_REF][START_REF] Costa | Origin and transport of trace metals deposited in the canyons off Lisboa and adjacent slopes (Portuguese Margin) in the last century[END_REF] into these productive ecosystems containing important stocks of commercially important fishes. Capbreton Canyon is located nearby the coast, with important urban and agricultural activities.

Indeed, since the Capbreton canyon is connected to the Adour estuary, its geomorphology and the local currents lead to transfer particles and micropollutants further, amplified during storm events (e.i. turbidic currents) [START_REF] Mulder | A few monthsold storm-generated turbidite deposited in the Capbreton Canyon (Bay of Biscay, SW France)[END_REF][START_REF] Salles | Simulating the 1999 Capbreton canyon turbidity current with a Cellular Automata model[END_REF][START_REF] Brocheray | 2000years of frequent turbidite activity in the Capbreton Canyon (Bay of Biscay)[END_REF]. For instance, in the Capbreton canyon sediments collected in the first 25 km from the coast, UV filters and synthetical musks have been detected where the highest concentrations were observed in sediment collected at 25 km from the coast [START_REF] Azaroff | Priority and emerging micropollutants distribution from coastal to continental slope sediments: a case study of Capbreton Submarine Canyon (North Atlantic Ocean)[END_REF]. This trend was also observed for mercury compounds [START_REF] Azaroff | Mercury and methylmercury concentrations, sources and distribution in submarine canyon sediments (Capbreton, SW France): implications for the net methylmercury production[END_REF]. Additionally, wastewater treatment plants (WWTPs) are known to be an important pathway for introduction of ECs both through the treated effluents and particles released in aquatic environment [START_REF] Miège | Fate of pharmaceuticals and personal care products in wastewater treatment plants -conception of a database and first results[END_REF][START_REF] Verlicchi | Occurrence of pharmaceutical compounds in urban wastewater: removal, mass load and environmental risk after a secondary treatment-a review[END_REF]. In coastal areas, those WWTPs might be the main source of ECs in coastal and submarine sediments [START_REF] Azaroff | Priority and emerging micropollutants distribution from coastal to continental slope sediments: a case study of Capbreton Submarine Canyon (North Atlantic Ocean)[END_REF].

Microorganisms play a key role in ecological processes such as biogeochemical cycling and among them the carbon cycle and the organic compounds degradation. This bioremediation provides an important ecosystem service for the maintenance of the environment quality. The physicochemical properties of the sediment, such as organic carbon, grain size or pH, drive most of its interactions with the contaminants [START_REF] Nowell | Contaminants in stream sediments from Seven United States metropolitan areas: part I: distribution in relation to urbanization[END_REF]. The bioavailability for the dwelling benthic organisms of these ECs is dependent on the adsorption, desorption and transformation processes which are themselves under control of the biogeochemical parameters. Even at low concentrations, those ECs can be hazardous for the sediment-dwelling benthic organisms. It is widely accepted that their microbial co-metabolization could be the main degradation pathway leading to limited structural changes and incomplete mineralization (e.g. formation of degradation products). Consequently, the fates of ECs in submarine canyon sediments depend on physicochemical properties, and on the presence and activity of microorganisms Several studies were performed to explore bioremediation by microorganisms of priority contaminants such as polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyl (PCBs) [START_REF] Perelo | Review: In situ and bioremediation of organic pollutants in aquatic sediments[END_REF][START_REF] Duran | Role of environmental factors and microorganisms in determining the fate of polycyclic aromatic hydrocarbons in the marine environment[END_REF]. Few studies have shown the potential of bacteria and microbial consortia involvement in the ECs remediation [START_REF] Ncibi | Remediation of emerging pollutants in contaminated wastewater and aquatic environments: biomass-based technologies[END_REF]. Biotransformation potential of ECs in natural and oceanic environments is still unknown and it is urgent to better understand their fate in marine ecosystems [START_REF] Gavrilescu | Emerging pollutants in the environment: present and future challenges in biomonitoring, ecological risks and bioremediation[END_REF].

The aim of this present work was to study the degradation potentials of ECs such as synthetic musks (galaxolide, HHCB; tonalide, AHTN), UV filters (padimate O, OD-PABA; octocrylene, OC) and an antiepileptic (carmabazepine, CBZ) in marine sediments in order to highlight, for the first time their natural resilience in a submarine canyon sediments. Moreover, a specific focus was put on the isolation of pure strains exposed to these hazardous substances in order to estimate their capacities in biotransformation of these contaminants in natural ecosystems and more particularly in marine sediments

Material and methods

Study area and sampling strategy

The Capbreton Canyon (South-Eastern Bay of Biscay, NE Atlantic) begins 250 m from the coastline and reaches up to 3000 m water depth. In this study, we focused on two surface sediments (G10 and G14) sampled in the Capbreton Canyon area during the oceanographic cruise HAPOGE organized in July 2017. Surface sediments (0-10 cm) were sampled with Shipeck sampler grab. First sampling station was into the canyon (G10) and the second one was on the adjacent continental shelf (G14) at 4.4 and 9 km from the coast, and at 70 and 120 m of depth, respectively (Fig. 1). After collection, the sediment samples were placed into sterile polyethylene bags sealed and stored in the dark at 4 °C until slurry incubations in the laboratory (described further) within 48 h. In parallel, for each sample, contaminants analysis and geochemical parameters were analysed [START_REF] Azaroff | Priority and emerging micropollutants distribution from coastal to continental slope sediments: a case study of Capbreton Submarine Canyon (North Atlantic Ocean)[END_REF][START_REF] Azaroff | Mercury and methylmercury concentrations, sources and distribution in submarine canyon sediments (Capbreton, SW France): implications for the net methylmercury production[END_REF]. Briefly, G10 sediments were more muddy than those in G14 and were characterized with particulate organic matter of 1.60 % and 0.92 %, respectively and fine grain size (< 63 μm) of 76 % and 46 %, respectively.

Chemicals

Ethyl acetate (EtOAc) and methanol were of analytical grade and supplied by Sigma Aldrich (Saint-Louis, USA). Acetone (laboratory reagent. 99.5 %) used for cleaning the glassware was supplied by Fisher (Hampton, USA). Ultrapure water was obtained with a PURELAB Classic water purification system from Veolia (Paris, France). Reference standards of galaxolide (HHCB), octocrylene (OC), padimate O (OD-PABA) and carbamazepine (CBZ) were purchased from Sigma-Aldrich. Tonalide (AHTN) was purchased from LGC Standards (Molsheim, France). Internal standards musk xylene-d15 (MX-d15) (100 ng μL -1 in acetone) was purchased from LGC Standards and carbamazepine-d10 (CBZ-d10) (100 ng μL -1 in methanol) was purchased from Sigma Aldrich.

Fig. 2.

Design of experiments to determine natural resilience and microbial strains involved in the degradation of emerging contaminants (ECs): Galaxolide (HHCB), Padimate O (OD-PABA), Carbamazepine (CBZ), Tonalide (AHTN) and Octocrylen (OC). First step was the determination of in situ degradation constant rates from slurry sediments spiked with three ECs under biotic and abiotic conditions at 100 ppb (AHTN and OC were not monitored). Second step was the selection of strains from the last slurry sediments incubation (day 110) through enrichments performed under oxic/ anoxic and biotic/abiotic conditions at 25, 25, 28, 36 and 24 ppm of HHCB, AHTN, OD-PABA, OC and CBZ, respectively (corresponding to 100 μM). These enrichments were repeated three times (1, 2, 3) with liquid multipurpose medium at 20 g L -1 of NaCl, called MM 20 . Third step was strains isolation from the last enrichment (3) where microbial consortia were separated (spread plates) on agar MM 20 at same ECs exposition concentrations than previously (A). After one month of growth, colonies were twice isolated by streak plating (B, C). The last step was the characterisation of isolated strains through both their identification and the determination of their ECs degradation capacities. Isolated strains were identified by 16 s rDNA amplification and Sanger sequencing. ECs degradation tests were performed at 1 ppm of ECs using liquid MM 20 suppplemented with peptone (5 g L -1 ), yeast extract (2.5 g L -1 ) and glucose (5 g L -1 ), called MM 20 -PYG. * indicates that results are presented in the figures.

Determination of half life: sediment slurry incubations

In the laboratory slurry incubations spiked with HHCB, OD-PABA and CBZ were performed for 110 days in order to estimate the degradation potential (Fig. 2). Briefly, for each station (G10 and G14), a slurry was prepared by mixing fresh sediment with underlying water (50:50, w:w). Incubation experiments were performed in 10 mL glass tubes sealed with PTFE stoppers filled with 10 g of slurry. For all assays under abiotic and biotic conditions, HHCB, OD-PABA and CBZ were added independently, to have a final concentration at 100 μg g -1 . Initial time assays were stopped immediately by storing samples at -80 °C whereas incubated assays were placed in the dark at 14 °C (in situ temperature) and stopped at different elapsed times until 110 days. Slurries under abiotic condition (control), were twice sterilized for 20 min at 120 °C. All assays were performed in triplicate.

Based on the assumption that degradation reaction of these contaminants are of pseudo-first-order (1),biodegradation rate constants (k) were estimated with the following Eq. ( 2) :

Ln(C)= f(t) (1) k = Ln(C 0 /C t ) t (2)
Where C t is the concentration of the contaminant at the kinetic time t; C 0 is the initial contaminant concentration; k is the biodegradation rate constant and t is the time. Then, half time reactions were also determined by the following Eq. ( 3):

t 1/2 = Ln(2)/k (3)

Enrichment experiments in liquid medium

In order to do a selective enrichment of microbial populations potentially involved in the ECs degradation, 0.5 g of oxic (surface, 0.1 cm, clear yellow color) and 0.5 g of anoxic (dark color) sediments (Fig. 2) were subsamples from the last days of sediment slurry incubations and were diluted with modified multipurpose medium [START_REF] Widdel | Gram-negative mesophilic sulfate-reducing bacteria[END_REF] at 20 g L -1 of NaCl, called MM 20 . The medium contained : 1 L of MilliQ water, 1 mL of SL 12 , 1 mL SeTg, HEPES 10 mM, yeast extract 0.1 g L -1 , 20 g NaCl, 3 g MgCl 2 6H 2 O, 0.15 g CaCl 2 H 2 O, 0.25 g NH 4 Cl, 0.5 g KCl. pH was ajusted at pH = 7.5 before sterilization and autoclave (20 min at 120 °C). After autoclaving, 1 mL of solution V7 (vitamins) and KH 2 PO 4 (final concentration of 0.2 g L -1 ) were added with a syringe and a cellulose nitrate 0.2 μm filter (Fisher).

While we determined the half life for 3 ECs, the enrichment experiments in liquid medium were performed for 5 ECs, HHCB, AHTN, OD-PABA, OC and CBZ. Since AHTN has the same molecular mass than HHCB but with a different chemical structure (Table 1), we performed the enrichment experiment for both from the HHCB sediment slurry incubation. Then, OC was added to this list due to its occurrence in coastal sediments [START_REF] Combi | Distribution and fate of legacy and emerging contaminants along the Adriatic Sea: a comparative study[END_REF][START_REF] Mitchelmore | Occurrence and distribution of UV-filters and other anthropogenic contaminants in coastal surface water, sediment, and coral tissue from Hawaii[END_REF]. ECs were added independently at 100 μM final concentration (corresponding to 25, 25, 28, 36 and 24 ppm, respectively) according previous experiments [START_REF] Vallecillos | Degradation of synthetic fragrances by laccase-mediated system[END_REF][START_REF] Balk | Environmental risk assessment for the polycyclic musks AHTN and HHCB in the EU: I. Fate and exposure assessment[END_REF][START_REF] König | Reductive transformation of carbamazepine by abiotic and biotic processes[END_REF][START_REF] Sauvêtre | Metabolism of carbamazepine in plant roots and endophytic rhizobacteria isolated from Phragmites australis[END_REF]. These ECs have low solubility in water (Table 1), and they were added first into the tube with organic solvent (EtOAc, or methanol for CBZ) and evaporated at ambient temperature, under microbiological safety cabinet (MSC). This step was performed in order to release organic solvent, a potential carbon source [START_REF] Delgado-Moreno | Innovative application of biobed bioremediation systems to remove emerging contaminants: adsorption, degradation and bioaccesibility[END_REF]. Enrichment was performed in an agitator (120 rpm) [START_REF] Vallecillos | Degradation of synthetic fragrances by laccase-mediated system[END_REF] under oxic and anoxic conditions, at 27 °C for one month. This enrichment was repeated three times, with each time a dilution at approximatively 1/11 of the previous assay. Briefly, under MSC, micropollutant was added in a falcon tube (50 mL) or penicillin tube (100 mL) (for oxic and anoxic conditions, respectively). Then 5.5 mL of the MM 20 were added before the addition of 0.5 g of anoxic and oxic sediments from the slurry incubations (1 st step, Fig. 2). After one month in the agitator, new tubes were prepared with ECs by using the same protocol. Then 5 mL of MM 20 were added and inoculated with 0.5 mL of the previous enrichment, corresponding to a 1/11 dilution (2nd and 3rd steps, Fig. 2).

Strains isolation experiments

After the third enrichment step, in order to isolate the strains previously selected and enriched, solid medium of MM 20 (with 20 g L -1 of agar bacteriologic, previously rinsed 3 times with ultrapure water) were prepared in Petri plates. Under MSC, micropollutants were added (274,274,306,498 and 254 μL of HHCB, AHTN, OD-PABA, OC and CBZ at 1000 ppm, respectively) at the surface of the solid medium and organic solvent was evaporated. Then, inocula of the last enrichments were spread at the surface of the solid medium with sterile inoculator and sealed with parafilm before incubation for 1 month, at 27 °C to favour the growth of colonies. After one month, colonies were selected and isolated by streak plates as described (A, B, C steps, Fig. 2). Then all strains isolated were stored at -80 °C in sterile LB (20 g L) supplemented with NaCl (20 g L -1 ), and glycerol 30 % (v:v).

Identification of isolated strains: DNA extraction, 16 s rDNA gene amplification, sequencing and phylogenetic analysis

DNA was amplified from isolated strains after growth in LB Lennox medium. Amplification of the 16S rRNA gene was done with the universal primers 63 F (5′-CAG GCC TAA CAC ATG CAA GTC-3′) and 1387R (5′-GGG CGG WGT GTA CAA GGC-3′) [START_REF] Marchesi | Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA[END_REF]. PCR amplification was performed using ampliTaq Gold® 360 master mix (Applied Biosystems, CA, USA), 0.2 μM of each primer and 1 μL of strain. PCR cycling was as following : after 10 min of initial denaturation at 95 °C (lysis of cells), 35 cycles of 40 s denaturation at 95 °C, 40 s annealing at 58 °C and 60 s elongation at 72 °C with 7 min final elongation at 72 °C. Amplicons were sequenced by SANGER sequencing at GATC (Köln, Germany). Sequences were trimmed with ChromasPro (Technelysium software) and were aligned with MUSCLE [START_REF] Edgar | MUSCLE: multiple sequence alignment with high accuracy and high throughput[END_REF].A tree was generated using MEGA X software [START_REF] Kumar | MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms[END_REF], with the Maximum Likelihood method and Tamura-Nei model [START_REF] Tamura | Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees[END_REF] (with n replication boostraps = 500). Phylogenetic analysis were processed with NCBI (https://www.ncbi.nlm.nih. gov/) and corresponding reference type strains as defined by the bacteriological code of nomenclature for Prokaryotes (http://www. bacterio.net/-classifphyla.html). Sequences are archived in GenBank under accession numbers MT658667 to MT658707.

In parallel, in order to compare with the isolated strains, DNA of in situ samples and from the final kinetic time of the slurry incubations were studied to determine the global diversity. DNA was extracted from frozen sediments with the QIAGEN DNeasy Powersoil kits (Qiagen Inc., Netherlands) according to the manufacturer's instructions. Diversity of the 16S rDNA were determined by sequencing the V4-V5 hypervariable regions of the 16S rDNA with universal primers V4-515 F (5′ GTGY-CAGCMGCCGCGGTA 3′) and V5-928R (5′-ACTYAAAKGAATTGRCG-GGG 3′) [START_REF] Parada | Every base matters: assessing small subunit rRNA primers for marine microbiomes with mock communities, time series and global field samples[END_REF][START_REF] Walters | Improved bacterial 16S rRNA gene (V4 and V4-5) and fungal internal transcribed spacer marker gene primers for microbial community surveys[END_REF][START_REF] Wang | Conservative fragments in bacterial 16S rRNA genes and primer design for 16S ribosomal DNA amplicons in metagenomic studies[END_REF]. PCR was performed using ampliTaq Gold® 360 master mix (Applied Biosystems, CA, USA), 0.5 μM of each primer and 3 ng of extracted DNA. PCR cycling was as following : after 10 min of initial denaturation at 95 °C, 30 cycles of 30 s denaturation at 95 °C, 30 s annealing at 60 °C and 40 s elongation at 72 °C with 7 min final elongation at 72 °C. Amplicons were sequenced using MiSeq 250-pair-end technology (Illumina, CA, USA) with V3 kit version, in Get-plage sequencing platform (INRA, Toulouse, France). Data were preprocessed using Galaxy FROGS pipeline [START_REF] Escudié | FROGS: Find, Rapidly, OTUs with Galaxy Solution[END_REF]. Chimera and PhiX reads were removed, Operational Taxonomic Units (OTUs) clustering, after a de-noising step allows building fine clusters with minimal differences, with an aggregation distance equal or above 3. Data were normalized with the minimum number of reads. Taxonomic affiliation was performed using the Silva database v.128 [START_REF] Pruesse | SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB[END_REF]. Sequences data in situ and from the final kinetic time of the slurry incubations have been deposited in Genbank under the accession number PRJNA608532 and PRJNA640934, respectively.

Experimental ECs degradation test

Isolated strains were re-cultivated in medium MM 20 -PYG (MM 20 medium supplemented with peptone, 5 g L -1 , yeast extracts, 2.5 g L -1 and glucose at 5 g L -1 ) à 37 °C for 90 h [START_REF] Popa | Potential of newly isolated wild Streptomyces strains as agents for the biodegradation of a recalcitrant pharmaceutical, carbamazepine[END_REF]. Optical density (DO) at 600 nm was measured during the growth until the maximum growth using spectrophotometer (Spectronic 20). Degradation tests were performed in glass tubes with contaminants (HHCB, AHTN, OD-PABA, OC and CBZ) evaporated [START_REF] Delgado-Moreno | Innovative application of biobed bioremediation systems to remove emerging contaminants: adsorption, degradation and bioaccesibility[END_REF]) to be at 1 ppm final concentration, with 1 mL of the cultivated strains, 9 mL of MM 20 -PYG at 27 °C in agitator (120 rpm) [START_REF] Vallecillos | Degradation of synthetic fragrances by laccase-mediated system[END_REF][START_REF] Popa | Potential of newly isolated wild Streptomyces strains as agents for the biodegradation of a recalcitrant pharmaceutical, carbamazepine[END_REF]. 10 μL of micropollutant at 1000 ppm were added with sterile syringe and PTFE filter 0.2 μm (Fisher). Degradation was stopped when DO 600 reached the maximum value. For initial and final times, chemical analysis were performed (in triplicates) to evaluate the ECs degradation capacity of the isolated strains under controlled conditions (described below).

ECs analysis

HHCB, AHTN, OD-PABA, OC and CBZ were extracted by a liquid:liquid extraction, for both the second enrichment in liquid medium (3) and the ECs degradation test (Fig. 2) at the initial and final time of each experiment. Briefly, 10 and 2 mL of EtOAc were added directly into the essay tube before to vortex for 1 min, respectively. Then, 20 μL of the supernatant was diluted with EtOAc (960 μL) in GC-vial and spiked with 20 μL of an internal standard mixture at 10 μL ng -1 (CBZ-d10 and MX-d15) before analysis. Extracts were analysed by 7890A gas chromatograph coupled with 5975C mass spectrometer (GC/MS) with an Electron Ionization (EI) source using a Large Volume Injection (LVI) (Agilent Technologies). The GC/MS system was equipped with a single taper ultra-inert liner with glass wool and a HP-5MS UI capillary column (30 m length ×0.25 mm diameter and 0.25 μm film thickness). Carrier gas was helium (He) with a purity greater than 99.999 % (Linde). Separation was performed at a constant He flow of 1.5 mL min -1 , and the GC oven temperature was programmed to hold at 50 °C for 3 min, then increase at 25 °C min -1 until 195 °C (hold 1.5 min), then 8 °C min -1 until 265 °C (hold 1 min) and 20 °C min -1 until 310 °C (hold 5 min). Injection volume was 20 μL in splitless mode. Instrument control, data acquisition and data treatment were performed using Agilent Chemstation software. Quantification was carried out in Selected Ion Monitoring (SIM) mode, selecting two characteristics fragments ions for each compound. Calibrations with the target analytes (HHCB, AHTN, OD-PABA, OC and CBZ) were used to quantify target compounds. A six-point calibration curve was performed in EtOAc spiked with increasing pollutants concentration levels ranging from 0 to 500 μg L -1 as well as internal standards at 50 μg L -1 transferred into GC vials (analytical performances are compiled in Table S1, supporting information). Recoveries achieved for all target ECs in MM 20 medium spiked at 100 μg L -1 ranged from 91 % to 102 % for musk and UV-filter compound and was at 74 % for CBZ (Supplementary information, Table S1).

Results and discussion

Degradation of HHCB, OD-PABA and CBZ in sediment slurry incubations

Based on kinetic degradation for 110 days under both abiotic and biotic conditions (Supplementary information, Fig. S1), degradation rate constants, k, were estimated from slurry incubations under biotic condition (Table 2). While CBZ in both sediment samples exhibited similar rates (0.0035 ± 0.0004 and 0.0029 ± 0.0001), HHCB and OD-PABA exhibited different rates of degradation according the location (e.g. G10 and G14) with a factor around 2 (Table 2). This suggested that ECs degradations were depending on the geochemical parameters of the sediments (e.i. grain size, organic matter content). For instance, a previous study demonstrated that ECs have a strong affinity for particles [START_REF] Li | Sorption and degradation of selected organic UV filters (BM-DBM, 4-MBC, and OD-PABA) in laboratory water-sediment systems[END_REF], as observed with sludge in wastewater treatment plants [START_REF] Liu | Occurrence and removal of benzotriazoles and ultraviolet filters in a municipal wastewater treatment plant[END_REF], suggesting those parameters and as well as environmental factors have to be taken in consideration to understand the fate of ECs in the ocean. Estimation of the half life time of reaction for these ECs indicated that the UV filter OD-PABA was between 21 ± 1 and 44 ± 4 days (Table 2). These results are similar with another UV filter degradation (i.e. EH-DPAB) estimated in marine sediments microcosm with a t 1/2 ranging from18 to 50 days [START_REF] Volpe | Biodegradation of UVfilters in marine sediments[END_REF]. Nevertheless, half life reaction for synthetic musk HHCB and anti-epileptic CBZ reached around 8 months and 7 months, respectively, indicated the high persistence of these contaminants in natural marine sediments compare to the UV filter OD-PABA (Table 2). Lack of data on HHCB and CBZ degradation

Table 2

First-order rate constants k (in days -1 ) for degradation of HHCB, OD-PABA and CBZ in the slurry sediments incubations under biotic condition (in triplicate). Pearson determination coefficients ranged from 0.76 to 0.98 and p-values < 0.05 for estimation of k. The corresponding half-lives (t 1/2 , in days) were calculated as ln(2)/k. G10 and G14 correspond to canyon surface sediment and continental shelf sediment, respectively. potentials in marine sediments does not allow comparison with these present data. Nevertheless, these preliminary results demonstrate the high persistence of these ECs in natural environment. In sediments, ECs may be associated to particulate matter, establishing equilibrium relations in the water-sediment interface, limiting their bioavailability. ECs remobilisation depends both of geochemical characteristics variations of the sediment, the overlying water column and pore water. Thus, ECs persistence in environment depend on coupling of abiotic and biotic factors (Lee [START_REF] Wolfe | New perspectives in aquatic redox chemistry: abiotic transformations of pollutants in groundwater and sediments[END_REF][START_REF] Jeon | Aerobic bacterial catabolism of persistent organic pollutants -potential impact of biotic and abiotic interaction[END_REF], such as physicochemical properties of the ECs, environmental factors (e.i. temperature [START_REF] Lange | Occurrence and fate of synthetic musk fragrances in a small German river[END_REF], pH [START_REF] Perelo | Review: In situ and bioremediation of organic pollutants in aquatic sediments[END_REF], redox processes [START_REF] Jeon | Coupling microbial catabolic actions with abiotic redox processes: a new recipe for persistent organic pollutant (POP) removal[END_REF] photolysis process [START_REF] Chiron | Photodegradation processes of the antiepileptic drug carbamazepine, relevant to estuarine waters[END_REF][START_REF] Vogna | Kinetic and chemical assessment of the UV/H2O2 treatment of antiepileptic drug carbamazepine[END_REF]) and the presence of microbial communities able to degrade these ECs [START_REF] Onesios | Biodegradation and removal of pharmaceuticals and personal care products in treatment systems: a review[END_REF]. It was also demonstrated that bioturbation or resuspension improved oxidation of anoxic sediments leading an increase of lability of ECs which can be more available for degradation processes [START_REF] Booij | Release rates of chlorinated hydrocarbons from contaminated sediments[END_REF]. Additionally, chemical characterization of the collected sediments revealed that the three-targeted ECs were not found at concentration detectable with the analytical method used [START_REF] Azaroff | Priority and emerging micropollutants distribution from coastal to continental slope sediments: a case study of Capbreton Submarine Canyon (North Atlantic Ocean)[END_REF]. The removal potential was observed under both abiotic and biotic conditions with percentage of removal for each compounds, HHCB, OD-PABA and CBZ, ranging from 37.5%-51.4%, from 50.1%-98.5% and from 11.6%-49.6%, respectively (Fig. 3). Results of HHCB and CBZ slurry incubations suggested that the degradation was mainly driven by an abiotic process (except for G10 exposed with CBZ). Removal potential of OD-PABA under biotic condition was 2 times higher compared to the abiotic condition, suggesting that micro-organisms in these sediments improved the degradation of this UV filter compound (Fig. 3).

Enrichment in liquid medium

Results of the second enrichment in liquid medium (Fig. 4) demonstrated a higher removal in oxic conditions than under anoxic conditions for the five ECs studies (e.i. HHCB, AHTN, OD-PABA and CBZ). These ECs are well known to be observed in WWTP, where CBZ is frequently detected in the liquid effluents while the other contaminants, due to the very hydrophobicity, occur in sludges or remain through membrane post-treatment [START_REF] Krzeminski | Occurrence of UV filters, fragrances and organophosphate flame retardants in municipal WWTP effluents and their removal during membrane post-treatment[END_REF]. Although new advanced treatment processes, such as activated carbon adsorption, advanced oxidation processes, reverse osmosis or membrane bioreactors, are developed to increase the micropollutant removal of the WWTPs to struggle against the spread of micropollutants in aquatic environment, they are still observed in the WWTP effluents [START_REF] Luo | A review on the occurrence of micropollutants in the aquatic environment and their fate and removal during wastewater treatment[END_REF]. Those results suggested that oxic processes could improve the ECs removal and limit their spread from the WWTP sewage. Moreover, higher removals under abiotic anoxic conditions than biotic were observed. It is likely due to a higher ECs adsorption on cells present in biotic condition leading to trap ECs and reduce the quality of the liquid extraction [START_REF] Bell | Removal of hazardous organic pollutants by biomass adsorption[END_REF][START_REF] Tsezos | Comparison of the biosorption and desorption of hazardous organic pollutants by live and dead biomass[END_REF]. Additionally, the high concentrations (ppm level) used for the selection of strains in these enrichments could inhibit microbial activity, thus could have mask their ECs removal capacities under anoxic condition [START_REF] Barra Caracciolo | Pharmaceuticals in the environment: biodegradation and effects on natural microbial communities. A review[END_REF].

Under oxic condition, similar removal potentials were observed for the abiotic and biotic condition, except for OC and CBZ (Fig. 4). That may indicate a potential involvement of microorganisms in the OC removal. Contrary to OC, CBZ removal was observed only in abiotic anoxic condition, likely due to the same explanations suggested previously for the removal potential under anoxic condition (e.i. adsorption on cells and/or high exposition concentrations). These results led to continue the study of ECs degradation with the isolated strains under oxic conditions.

Nevertheless, even though an efficient removal of ECs under oxic condition was observed, metabolites were not studied. Indeed, degradation of ECs to metabolites have been observed for CBZ [START_REF] König | Reductive transformation of carbamazepine by abiotic and biotic processes[END_REF], HHCB [START_REF] Vallecillos | Degradation of synthetic fragrances by laccase-mediated system[END_REF] and also for others UV filters [START_REF] Volpe | Biodegradation of UVfilters in marine sediments[END_REF]. These results highlighted the complexity of ECs removal under experimental conditions as well as in WWTPs, where their efficiency are dependant of both physical, chemical and biological processes but also to identify metabolites in further studies. ,2,3,4 and 5 included 10 (20,22,6,7,8,11,3 (33,35,4 (17,18,6 (45,46,47, and3 (41, 42, 43 OC-G10) isolated strains, respectively.

Isolated strains involved in ECs degradation

While the degradation of complex organic substances can occurs both under anaerobic and aerobic conditions, it is well known that their degradation rates can be higherunder aerobic condition [START_REF] Biel-Maeso | Sorption and degradation of contaminants of emerging concern in soils under aerobic and anaerobic conditions[END_REF][START_REF] Ying | Biological degradation of triclocarban and triclosan in a soil under aerobic and anaerobic conditions and comparison with environmental fate modelling[END_REF]. In accordance with this observation, isolation of strains and degradation test were consequently performed under aerobic condition. Twenty seven strains were isolated with the current method with 6, 8, 4, 4 and 5 strains constrained with HHCB, AHTN, OD-PABA, OC and CBZ, respectively (Supplementary information, Fig. S3). These strains were related to two families, dominated by the Firmicutes, followed by the Actinobacteria (Fig. 5 A B).

Within the Actinobacteria, Rhodococcus was the main bacteria genus isolated followed by Streptomyces, Mycobacterium and Micrococcus. Within the Firmicutes, only Bacillus-related strains were isolated (Fig. 5 A,B). Studies have shown that Actinobacteria were involved in degradation of some organic and inorganic compounds. For instance, Rhodococcus strains are involved in the removal of pharmaceutical compounds [START_REF] Ivshina | Biodegradation of emerging pollutants: focus on pharmaceuticals[END_REF], tetrabromobisphenol A (TBBPA) [START_REF] Xu | Transformation of tetrabromobisphenol A by Rhodococcus jostii RHA1: effects of heavy metals[END_REF] and dominant in naphthalene degrading enrichments [START_REF] Rhee | Detection of genes involved in biodegradation and biotransformation in microbial communities by using 50-Mer oligonucleotide microarrays[END_REF]. It has also been demonstrated that the members of the genus Streptomyces produce laccase (i.e. ligninolytic enzymes) that can be involved in the removal of recalcitrant and ECs [START_REF] Majeau | Laccases for removal of recalcitrant and emerging pollutants[END_REF]. It has also been demonstrated that members of the genus Streptomyces were involved in the CBZ biodegradation [START_REF] Popa | Potential of newly isolated wild Streptomyces strains as agents for the biodegradation of a recalcitrant pharmaceutical, carbamazepine[END_REF][START_REF] Popa Ungureanu | Response surface optimization of experimental conditions for carbamazepine biodegradation by Streptomyces MIUG 4.89[END_REF]. Finally, experiments performed from marine sediments demonstrated that Bacillus thuringiensis was a novel group participating to the removal of PAHs and pesticides [START_REF] Ferreira | Bacillus thuringiensis a promising bacterium for degrading emerging pollutants, process[END_REF].

Within these isolated strains, only eight exhibited a capacity to degrade contaminants in the conditions used (1 ppm, 5 days, 37 degrees in rich organic medium) with 1, 2, 3 and 2 strains for HHCB, AHTN, OD-PABA and OC, respectively (Table 3). Seven were related to the genus Bacillus and one was related to the genus Rhodoccocus (23 AHTN G14). While studies have demonstrated high bioremediation capacities by Actinobacteria, this study demonstrated high capacity of marine bacillus to degrade ECs. These results suggested that the isolated bacillus were able to remove OD-PABA with a capacity ranging between 34 % ± 19 % and 71 % ± 19 % (Table 3). These strains could be involved in the biotic degradation observed in the slurry incubation experiment (Fig. 3). Although CBZ slurry incubations experiments (G10) demonstrated a possible biotic degradation (Fig. 3), isolated strains did not exhibit a remediation for this pollutant under the conditions tested (Supplementary information, Fig. S3). Moreover, the t 1/2 of CBZ was estimated around 7 months, suggesting that the time should be expanded to observe a remediation capacity. Moreover, it is well known that in nature, the microbial consortia have to be considered to estimate biodegradation pollutant removal process [START_REF] Wu | Evaluating adsorption and biodegradation mechanisms during the removal of Microcystin-RR by periphyton[END_REF].Moreover, global microbial communities analysed in situ, in G10 and G14 demonstrated a relative abundance of the main phyla similar in both sediments (and similar to the final kinetic time of the slurry incubations, Supplementary information Fig. S2), with a dominance of Proteobacteria (relative abundance ranged between 47.9 and 49.4 %), followed by Bacteriodetes, Planctomycetes and Acidobacteria (Fig. 6). Although the Actinobacteria and Firmicutes were minors in situ (Fig. 6), representing less than 1% of the total abundance, phylogenic trees based the sequences of the isolated strains (Fig. 5 A,B) demonstrated that these isolated strains were related to the Actinobacteria and Firmicutes families. This suggests a good selection process of these bacterial groups in the presence of these ECs exposition with the isolation method used.In accordance with previous studies, the isolated bacterial strains from the Capbreton Canyon sediments could be good candidate to degrade ECs. Nevertheless, in order to estimate the natural resilience of those ECs, it is crucial also to take account both the biological (e.i. biodegradation, biosorption/adsorption), physical and chemical processes, highlighting further studies are needed to decipher their fate in oceans. It is difficult to extrapolate results based on pure strains to the field. The culture conditions used (medium, temperature, …) enhance some microorganisms and inhibit others. The role of co-metabolisms in microbial consortia is extremely important as well as the role of the uncultivated prokaryotes. But the objective of this study in fine to isolate strains on which we will be able to work on. The isolated strains can be a fantastic tool since they can be used to study the degradation products, to study the isotopic fractionation due to micro-organisms, or to study the genetic determinism of this pathway. These data could then be used as new tools for new environmental studies (compound specific isotopic fractionation, functional genes, ...).

Table 3

Removal capacity of isolated strain and their related affiliation based on phylogenetic tree (Fig. 5). % of removal is calculated relatively to the control removal and in triplicate, after 120 h of experimentation at 1 ppm exposition concentration for each emerging contaminants. The strains for which degradation were not determined are not shown (Fig. 5).

Conclusion

This work characterized the degradation kinetics for ECs (HHCB, OD-PABA and CBZ) from marine sediments under in situ conditions (biotic, anaerobic and temperature) thought slurry sediment incubations. The half life times obtained demonstrated that half time live of OD-PABA was lower than HHCB and CBZ, with t 1/2 ranging from few weeks to 8 months. This underlies that the ECs degradation in marine sediment is a long process and could be a high concern for the environment considering a chronical contamination from the coastal human activities (e.i. WWTPs sewage, agriculture…). Additionally, contrary to CBZ, different half lives were observed between muddy (G10) and sandy (G14) sediments for HHCB and OD-PABA. That indicates ECs degradation depends on the physicochemical properties of both chemicals and sediment.

Moreover, novel synthetic musks and UV filters degrading bacteria were isolated from marine sediment. Based on phylogenetic characteristics, microorganisms were mainly identified as Bacillus sp. dominated by Bacillus megaterium followed by Bacillus aquimaris and Bacillus vietnamensis. One other isolated strain which was able to degrade musk was related to Rhodococcus ruber. The isolated bacteria here, could have a strong potential to degrade a wide variety of organic pollutants (e.g. priority and emerging), such as synthetic musks and UV filters. Improving the knowledge about the microbial diversity in marine environment may lead to the development of technical tools with degradation abilities and high tolerance in the future but also highlight how microbial remediation contributes to the nature resilience in marine environment.

Fig. 1 .

 1 Fig. 1. Sediment sampling sites in the Capbreton Canyon (dark circle) and on the adjacent continental shelf (white circle). Isobath data from SEDIMAQ3 (Gillet 2012).

Fig. 3 .

 3 Fig. 3. HHCB, OD-PABA and CBZ removal potential (%) after 110 days of sediment incubation ()under biotic and abiotic conditions from Capbreton Canyon sediments (G10, and G14). Initial exposure concentration was 100 ppb. Data are mean ± SD of three replicates.

Fig. 5 .

 5 Fig. 5. Phylogenic trees of strains obtained after the aerobic isolation in solid medium with different ECs (AHTN, HHCB, OC, OD-PABA and CBZ) from sediments collected in the submarine Canyon of Capbreton (stations G10 and G14). Fig.5A and Fig.5B correspond to Actinobacteria and Firmicutes, respectively. Isolated strains are in bold, reference strains are in italic. The evolutionary history was inferred by using the Maximum Likelihood method and Tamura-Nei model. The bootstrap values areabove 50 % are shown next to the branches. The scale indicates nucleotides substitution. Groups 1, 2, 3, 4 and 5 included 10 (20, 21 AHTN-G10; 22, 27 6, 7, 8, 11, 3 (33, 35, 4 (17, 18, 6 (45, 46, 47, 48 CBZ-G14 and 50, 52 CBZ-G10) and 3 (41, 42, 43 OC-G10) isolated strains, respectively.

Fig. 6 .

 6 Fig. 6. Relative abundance of prokaryotic 16S rDNA gene sequences (MiSEQ) from sediments of G10 and G14. Data are average percentages ( ± SE). Categories representing > 1% are shown. Relative abundances of phyla from which the isolated strains belonged are also shown.

Table 1

 1 Characteristics of the emerging contaminants used in this study. Log K ow is octanol/water partition coefficient.

	Compound	Family	Formula	Molar mass g mol -1	Solubility in water mg L -1	log K ow	References
	Galaxolide (HHCB)	Musk	C 18 H 26 O	258.4	1.75	5.70	(Field et al., 2006)
	Tonalide (AHTN)	Musk	C 18 H 26 O	258.4	1.25	5.90	(Field et al., 2006)
	Padimate O (OD-PABA)	UV filter	C 17 H 27 NO 2	277.4	0.54	6.15	(Li, 2014)
	Octocrylen (OC)	UV filter	C 24 H 27 NO 2	361.5	0.36	6.88	(Rainieri et al., 2017)
	Carbamazepine (CBZ)	Pharmaceutical	C 15 H 12 N 2 O	236.3	17.7	2.45	EPA's EPI Suite TM

Acknowledgements

This work was supported by the European Union and the Adour Garonne Water Agency (Micropolit Project). Europe commits the New Aquitaine with the European Regional Development Fund.

Author statement

All person who meet authorship criteria are listed as authors, and all authors certify that they have participated sufficiently in the work to take public responsibility for the content, including participation in the concept, design, analysis, writing, or revision of the manuscript. Furthermore, each author certifies that this material or similar material has not been and will not be submitted to or published in any other publication before its appearance in the Journal of Hazardous Material Specific contributions made by each author (list the authors' initials) : R.G., A.A., designed research; M.M., R.G. acquired funding; A.A., R.G., M.M., C.M., C.G. collected data ; A.A., R.G., C.M.,C.G. analyzed data; A.A, R.G. wrote the paper, A.A., R.G., M.M reviewed and edited the paper. All authors have read and agreed with the published version of the manuscript.

Declaration of Competing Interest

The authors declare that they have no conflict of interest.

Appendix A. Supplementary data

Supplementary material related to this article can be found, in the online version, at doi:https://doi.org/10.1016/j.jhazmat.2020.123477.


Béatrice Jongy

Thomas Bernhard

Identität Cf Bernd Neumann

Rollenzwang Zur

R M Rilke

Les Carnets De Malte

Laurids Brigge

La filiation chez Thomas Bernhard, dans les récits autobiographiques

Selon la définition proposée par Philippe Lejeune 1 , c'est sur le développement de la personnalité que le récit autobiographique met l'accent, aux XVIIIe et XIXe siècles. Dans sa préface aux Confessions, Rousseau refuse tout héritage des "auteurs de [s]es jours : "De tous les dons que le ciel leur avait départis, un coeur sensible est le seul qu'ils me laissèrent 2 ". Dans la suite du récit, il met l'accent sur son cheminement biographique loin de tout cadre familial, de toute tradition sociale ou culturelle déterminantes. L'autobiographie devient donc, vers la fin du XVIII e et le début du XIX e siècles, le genre privilégié de l'individualisme bourgeois. L'individu bourgeois n'est plus guidé par une instance divine, féodale ou familiale, mais il est son seul guide au cours de son développement intellectuel 3 . Au XIXe siècle se produit en Europe une prise de conscience quant aux contraintes s'exerçant sur le sujet dans son entreprise autobiographique. De nouvelles approches épistémologiques des connaissances humaines, notamment dans l'historiographie, la génétique et la psychanalyse, révèlent la détermination du sujet par des forces qui lui échappent et dont il doit tenir compte. L'homme prend conscience de son individualité en même temps que des menaces qui pèsent sur elle. On voit apparaître dans le genre autobiographique la revendication de la singularité. Dans ce contexte, deux textes sont fondateurs : le Journal de Kafka (rédigé entre 1909 et 1923) et les Carnets de Malte Laurids Brigge, journal semi-fictif de Rilke rédigé entre 1904 et 1910. Or ces auteurs ont maintes fois affirmé la nécessité pour l'individu de s'affranchir de sa filiation pour devenir, selon le mot de Rilke, le "fils de personne" 4 . Quelques décennies plus tard, Thomas Bernhard manifeste une même méfiance (autrichienne ?) à l'égard de la famille associée au désir d'autogenèse, dans ses récits autobiographiques : L'Origine, La Cave, Le Souffle, Le Froid et Un Enfant, parus entre 1975 et 1982.

LA FILIATION EN ECHEC

Une famille de danseurs de corde

Chez Bernhard, la filiation est perçue la plupart du temps comme une malédiction.

tendances suicidaires, l'oncle qui a mis fin à ses jours et la mère dépressive, soumise au père tyrannique. Dans son autobiographie, l'écrivain dépeint à plusieurs reprises l'action destructrice du grand-père, Johannes Freumbichler, sur sa fille Herta. Celle-ci a passé son adolescence et sa jeunesse comme femme de chambre et cuisinière au service de familles bourgeoises suisses et autrichiennes pour venir en aide financièrement à ses parents.

Johannes en effet travailla toute sa vie à son roman et refusa d'occuper un emploi dévalorisant. Il imposait à sa famille des privations et attendait de sa femme et de sa fille, comme il l'écrit à la première en 1926, un "entier dévouement et sacrifice". Herta supporte très mal sa condition. La joie éprouvée à la naissance de son fils cède vite la place à la difficulté de s'occuper seule d'un enfant dans sa situation. Dans Un Enfant, Bernhard explique que sa mère vivait près de l'abîme, toujours tentée par le suicide 5 . Cet état dépressif se transmettra au fils. Le désir inconscient qu'a Johannes d'anéantir sa fille, celle-ci le reporte sur son enfant. Ainsi l'écrivain qualifie-t-il de "destructrices" 6 les intentions du grand-père sur sa fille, et quelques pages plus loin, de "dévastatrices" les paroles de sa mère sur lui 7 . Une malédiction pèse sur la généalogie familiale. Bernhard compare d'ailleurs, dans une longue phrase essoufflée, sa famille à des danseurs de corde survolant le gouffre 8 . Le jeune Thomas n'a pas le droit de parler de son père car le sujet est tabou. Dans son autobiographie, il ne le nomme qu'une seule fois 13 . La quête d'identité est désespérée, car il ne reste plus rien du père. Quand l'écrivain obtient une photographie par son grand-père paternel, sa mère la lui arrache et la jette dans le poêle. Thomas Bernhard ne sait rien de son père, et n'a jamais eu, dit-il dans Le Froid, que des présomptions 14 . Le tableau qu'en fait la mère est celui d'un menteur et d'un criminel, qui en outre a fait d'autres bâtards : "des créatures comme toi" 15 , assène-t-elle à son fils. Lui pourtant ne peut s'empêcher de chercher les traces d'un père auquel il ressemble, au grand désespoir de la mère. C'est là l'un des thèmes clé de la bâtardise (que l'on trouve également chez Violette Leduc) : la ressemblance à celui qui a rompu la transmission mais se trouve légataire malgré lui.

La bâtardise

La vindicte maternelle

La mère lui reproche son statut de bâtard. Dans Un Enfant, il rapporte la scène traumatique où Herta l'envoyait chercher les cinq marks que l'Etat payait pour son entretien, le premier du mois, avec cette remarque "afin que tu voies ce que tu vaux" 16 , et qu'il décrit comme une course en Enfer. Il évoque le désespoir, l'envie de mourir, la culpabilité ressentis enfant 17 . Le bâtard, véritable enfant du diable, est assimilé au péché. En outre, l'enfant est en situation d'échec scolaire 18 . Il est considéré comme un malheur, un semeur de brouille, un menteur 19 . Ces termes, comme tant d'autres, sont placés en italiques, car il s'agit de la voix de la mère qui hante l'enfant, et qui traverse les pages sous la forme du discours indirect libre.

Ce ressassement, note Bernhard, l'emprisonne dans son rôle de bâtard et de criminel. La répétition douloureuse laisse son empreinte et le nom du père est remplacé par des surnoms humiliants : semeur de désordre et pisse-au-lit. 20 Ce "nouveau titre, quasiment mortel" 21 , selon les mots de l'intéressé, fonctionne comme un second baptême. L'écrivain ne cesse de souligner le pouvoir mortifère des paroles maternelles qui le précipitent dans un abîme dont il n'est plus jamais sorti 22 . Herta considère son fils comme un animal indomptable. Au fur et à mesure qu'il grandit, elle fait preuve à son égard d'une violence croissante. Entre la mère et le fils, l'amour ne peut s'épanouir. Lors de la fugue du jeune Thomas en bicyclette, Herta désespérée a alerté le voisinage et la police 23 . Néanmoins, seule la protection du grand-père évitera le châtiment au fugitif. La mère est épuisée par le conflit, prisonnière et victime de Cette douleur reçue en partage, cette solitude existentielle, l'écrivain va pourtant les retourner comme une arme et faire triompher sa singularité.

L'INVENTION DE SOI

La mythographie D'abord, il choisit sa filiation : il sera l'héritier direct de son grand-père. "Les chemins que j'ai suivis ont déjà été parcourus par lui, mon grand-père", écrit-il dans La Cave [START_REF] Tb | Die Wege, die ich gegangen bin, waren von ihm, meinem Großvater, schon begangen[END_REF] . Il tient de Johannes l'esprit de contestation. Le portrait qu'il trace de ce dernier, celui d'un anarchiste opposé au commerce, à la bourgeoise, à l'Eglise, à la foi, à l'école, aux maîtres, aux femmes, est aussi un autoportrait. Dans Un Enfant, Thomas Bernhard montre que c'est le grand-père qui lui a appris le langage 25 . "Grand-père" est d'ailleurs le premier mot qu'il aurait prononcé 26 … Sa carrière d'écrivain est clairement présentée comme un héritage de Johannes, qui lui a légué la machine à écrire qu'il utilisera toute sa vie [START_REF] Tb | La Cave[END_REF] . A la mort de Freumbichler, Bernhard se met à lire ses livres 28 et à écrire. Il évoque ses débuts en écriture :

"Dès cette époque je m'étais réfugié dans l'écriture, j'écrivais et j'écrivais, je ne sais plus, des centaines et des centaines de poèmes, je n'existais que lorsque j'écrivais, mon grand-père, le poète était mort, maintenant moi, j'avais le droit d'écrire, maintenant moi, j'avais la possibilité d'écrire moi-même des poèmes, maintenant je me risquais…" [START_REF] Tb | Ich hatte mich schon zu dieser Zeit in das Schreiben geflüchtet, ich schrieb und schrieb, ich weiß nicht mehr, Hunderte, Aberhunderte Gedichte, ich existierte nur, wenn ich schrieb, mein Großvater, der Dichter, war tot, jetzt durfte ich schreiben, jetzt hatte ich die Möglichkeit, selbst zu dichten, jetzt getraute ich mich[END_REF] C'est paradoxalement à travers Johannes, et grâce à sa mort, que peut se rétablir la filiation avec la mère, devenue substitut du disparu : "Sa nature m'était apparue clairement tout à coup, elle était la plus proche de celle de mon grand-père…" [START_REF] Tb | Ihr Wesen, war mir aufeinmal deutlich geworden, war dem meines Großvaters am nächsten[END_REF] Thomas Bernhard s'empare également de l'héritage paternel. D'une part en tant que criminel, comme ce père qui fait peur à toute la famille, et dont il dit qu'"il fut toute sa vie le troublefête" [START_REF] Tb | war der lebenslängliche Spielverderber[END_REF] . Or il utilise une expression très proche (d'ailleurs traduite par le même terme dans l'édition française) de La Cave : "un pareil trouble-fête je l'ai été toute ma vie" 32 . D'autre part en tant qu'écrivain, puisque la mère dit que le père avait l'art de faire de belles phrases 33 Il est vrai que l'autobiographie bernhardienne est le lieu d'une constante mise en scène de soi. L'incipit d'Un Enfant, où l'auteur raconte son escapade à vélo, est construit comme un petit drame en quatre actes : Le triomphe du héros (le trajet vers Salzbourg), la catastrophe (la chute), le réconfort (la nuit chez la paysanne), la glorification du coupable (l'arrivée chez les siens). A la faveur un véritable coup de théâtre orchestré par l'enfant devant Chorchi puis par le grand-père devant la mère, l'échec de cette escapade à vélo est transformé en un acte de bravoure48 . Bernhard dramatise le récit par l'usage des répétitions lexicales et parallélismes syntaxiques, très souvent supports d'une gradation sémantique.

"Cette décision audacieuse, cette décision fatale" [START_REF]kühnen", "verhängnisvollen Entschluß[END_REF] , écrit-il à propos de son projet d'excursion à Salzbourg. Le dernier adjectif vise en outre à produire une attente. La simple promenade est d'ores et déjà présentée par le prologue comme une tragédie. Le même type de gradation réapparaît plus loin, donnant à la narration un tour épique : "le très haut degré et même le degré suprême" [START_REF]höchsten, ja den allerhöchsten Grad[END_REF] . L'hyperbole est la seconde figure privilégiée par l'auteur pour ériger sa statue de héros : "pour la toute première fois", "avec la plus haute prétention" [START_REF]mein Vergehen oder gar Verbrechen[END_REF] . "Mon délit ou même mon crime", tels sont les termes qualifiant cet acte de désobéissance fondateur, annonciateur de l'opposition radicale qui sera la sienne à l'adolescence, et qui le poussera à aller "dans le sens opposé" [START_REF]Expression récurrente dans La Cave[END_REF] , quittant le lycée et les études bourgeoises pour une place d'apprenti, premier pas vers la carrière d'écrivain. La rupture du geste est figurée par les nombreux parallélismes : "Sans demander la permission", "sans avoir dit un mot." [START_REF] Tb | ohne um Erlaubnis zu bitten[END_REF] Le suspense de la scène est induit par l'hypotaxe. Bernhard place en tête de phrase toutes les subordonnées, et finit par la principale. Autrement dit, il énonce d'abord les circonstances, accentuant cette tendance de la langue allemande à finir sur le thème, comme en témoigne le passage suivant : "Ohne einem einzigen dafür zuständigen Menschen ein Wort gesagt zu haben, verließ ich auf der luftigen Höhe des Waffenrades und des damit verbundenen Vergnügens den Stadtplatz, um schliesslich in der sogennanten Au und dann in der freien Natur Richtung Salzburg die Räder laufen lassen." [START_REF] Tb | Sans avoir dit un mot à une seule personne compétente, jouissant de la hauteur aérée du vélo et du plaisir lié à cette situation, je quittai la Grand-Place pour finalement faire aller mes roues dans ce qu'on appelait la Prairie puis en pleine nature, direction Salzbourg[END_REF] Les questions rhétoriques empruntant à l'art oratoire judiciaire lui permettent, à la manière d'un Cicéron, de mettre en scène le procès dans le discours : "Qui, sinon moi, pourrait bien réussir à monter à bicyclette pour la toute première fois et à prendre le large, en outre avec la plus haute prétention : aller à Salzbourg !" [START_REF] Tb | Wem, außer mir, gelingt es schon, zum aleererstenmal auf das Rad zu steigen und auf und davon zu fahren, und noch dazu mit dem höchsten Anspruch, nach Salzburg![END_REF] L'acmé de cet autoportrait est l'hyperbole qui fait du cycliste un véritable homme volant, le "souverain du monde" 56 . L'autobiographie permet ici de se métamorphoser en un "il" glorieux, de s'octroyer une nouvelle identité. Dans d'autres passages, il n'hésite pas à se présenter comme un enfant exceptionnel, tournant en avantage ce qui était un défaut. Dans la scène du trajet vers le foyer pour enfants difficiles, il fait de sa différence -fruit du malheur -une supériorité. Les autres enfants sont médiocres, on confond leurs visages, lui se dépeint comme unique, et leur raconte l'histoire de sa bâtardise comme une histoire héroïque. Son origine intellectuelle même, discriminante, est valorisée. Son grand-père est écrivain, et lui connaît des mots que les autres ne connaissent pas, comme le terme "manuscrit" [START_REF]Manuskript[END_REF] . Surtout, il reprend à son compte le geste de l'exclusion : quand l'infirmière prend ses mains dans les siennes, il ne cède pas à la tendresse, mais manifeste répulsion et dégoût 58 . Il continue de tracer pour le lecteur l'image de l'indomptable.

Le salut par l'autobiographie

L'autobiographie est à la fois le lieu d'une enquête et d'un procès. Bernhard questionne ses origines, le silence fait autour de son père, cette démolition systématique dont il a été victime. Il lui faut "rechercher [s]on origine", "éclaircir" "le fourré touffu", ce que le narrateur du Froid appelle son "dévergondage spéculatif" [START_REF] Tb | [END_REF] . L'autobiographie vient pallier la défaillance de celui qui n'a osé questionner les siens tant qu'ils étaient vivants 60 , et consiste à défaire "le paquet et en plus devant des témoins, comme à présent, en déballant ces phrases grossières et brutales", sans aucune pudeur! ! ! 61 . Ainsi, dans Un Enfant, il raconte comment, quarante ans après, il revient voir le foyer pour enfants difficiles 62 . Il apprend alors le vrai nom du lieu, et dénonce l'éternelle comédie qui a transformé ce foyer nazi en foyer catholique. Bernhard prend sa revanche par son intelligence et sa capacité d'analyse. Ce retour courageux sur ses pas est la métaphore de l'autobiographie, enquête en même temps que quête de soi. Il s'agit de retrouver l'enfance pour poser son identité, se légitimer, mais aussi pour percer à jour cette société du mensonge, arracher le masque. C'est une sorte de Nuremberg personnel qu'accomplit ici l'auteur. L'auto-légitimation du bâtard se double connues de tous. Quant aux imprécations maternelles, elles sont à tout jamais conjurées. Le ressassement du fils remplace celui de la mère. La narration faite par l'adulte lui permet de triompher définitivement de la voix destructrice, qu'on entend en italiques, sertie dans le texte de Bernhard, étouffée par lui.

  Bernhard est né bâtard. Or, dans les civilisations occidentales, fondées sur le couple, la distinction entre enfants légitimes et naturels est prégnante. Herta doit accoucher en secret aux Pays-bas. Dans Un Enfant, Thomas Bernhard écrit que sa naissance n'aurait pas été possible à Henndorf, où résidait la famille. En réalité, les craintes des parents sont sans doute injustifiées. Le grand-père a vécu en concubinage avec sa future femme qui a abandonné pour lui son mari et ses enfants, la tante Rosina avait un fils naturel, ce qui n'était pas rare dans la région. La réaction bourgeoise de la famille est démesurée. Aussi l'enfant ne bénéficie-t-il pas d'un bon accueil au début de son existence. Pendant ses premiers mois, la mère doit déménager plusieurs fois, enchaînant les emplois pénibles de cuisinière. L'enfant est mis en nourrice, puis dans un foyer aux tarifs élevés. Herta doit choisir entre son père et son fils, car arrivent de Vienne des lettres de chantage. Elle place alors Thomas sur un bateau-maison, et ne lui rend visite qu'une fois par semaine. L'enfant ne sera accueilli par ses grands parents qu'à l'âge de neuf mois, alors qu'Anna est sage femme de profession et qu'ils sont nourris par leur fille… Herta doit repartir travailler en Hollande, et lorsqu'elle revient enfin à Vienne comme domestique, elle n'est pas logée chez ses parents, mais chez ses 5 Thomas Bernhard (abrégé en TB), Un Enfant, in L'Origine -La Cave -Le Souffle

…

  Le rôle de l'autobiographie chez Bernhard (comme chez Rousseau) est de convaincre le lecteur de son élection. Tout, dans le récit, vise à faire de lui un être extraordinaire. Comme tout héros, il doit avoir une généalogie exemplaire. ThomasBernhard s'inscrit dans une filiation héroïque, comme l'a montré Louis Huguet 34 : celle du grand-père qui a refusé l'héritage de petit épicier et d'aubergiste, et fait voeu de pauvreté pour l'art. Il aurait aperçu sa fiancée Ana depuis la fenêtre du séminaire, se serait enfui et l'aurait enlevée 35 à l'homme auquel elle était mariée, de trente ans son aîné. On retrouve ici, souligne Hans Holler 36 un des thèmes de prédilection de Bernhard : l'observateur par la fenêtre, et le chiffre 3 à caractère mystique. En réalité, l'écart d'âge entre Ana et son mari était moins important, Johannes a épousé la jeune femme à l'église, et exerçait la profession de coupeur dans une maison de confection. Selon son petit-fils, il aurait été immatriculé à la faculté et aurait eu des démêlés avec la police suisse. En fait il a été déchu de droits civils pour deux ans et interdit de séjour pour dettes. Toute la famille sort d'ailleurs grandie du portrait que fait l'écrivain. Si sa tante Maria a bien, comme il le dit, quitté le village pour Vienne où elle a épousé un peintre avec lequel elle est partie pour l'Amérique du Sud, celui-ci n'est pas devenu comme le prétend Bernhard une célébrité, mais il est mort inconnu. Surtout, Aloïs Zuckerstätter n'est plus, sous la plume de son fils, un séducteur de village, mais un génie 37 . A l'égard du père, Thomas Bernhard éprouve une certaine fascination, comme si ce geste de rupture -l'abandon, la fuite -, il pouvait le comprendre. Dans Le Froid, il raconte qu'Aloïs a incendié maison de ses parents avant de quitter définitivement son pays natal 38 . En réalité c'est un aïeul paternel qui a mis le feu à la maison pour toucher la prime d'assurances. Bernhard affirme tenir du père d'Aloïs que ce dernier aurait eu cinq enfants illégitimes 39 . Or il semble n'en avoir eu qu'un, la soeur Hilda. Cette représentation d'Aloïs en père de famille nombreuse l'apparente à Abraham, celui qui aurait pu sacrifier son fils, Thomas devenant ainsi une figure d'Isaac. Bernhard opère un déplacement : c'est l'oncle maternel Rudolf qui avait cinq enfants 40 . Enfin le mystère de la mort violente du père contribue au mythe. L'écrivain prétend qu'il aurait été fusillé dans des circonstances mystérieuses à la fin de la guerre. Il n'est pas jusqu'à l'oncle Rudolf qui ne se soit suicidé pour une raison élevée. La mère elle-même compare son fils à son frère, le génie de la famille, et reconnaît qu'elle a donné naissance à un enfant

L

  'enfant illégitime, victime d'exclusion, exclut à son tour toute la société dans un geste superbe. Il veut être la malédiction de sa famille et interrompre la chaîne de transmission. Il se décrit volontiers comme un ferment de ruine. Thomas Bernhard ne s'est pas marié, n'a pas eu d'enfant. Il vécut, platoniquement, avec son amie Hedwig Stavianicek, rencontrée en 1950, qui lui tint lieu de famille et dont il partage désormais la tombe. Comme Kafka, c'est aussi en tant qu'écrivain qu'il refuse de transmettre, mais seulement à ses compatriotes. Dans son testament, il interdit en effet la diffusion et la représentation de ses oeuvres en Autriche ("quelle que soit la forme de son État") pour les cinquante prochaines années 70 . S'il faut l'inscrire dans une lignée, c'est plutôt dans celle des écrivains -souvent autrichiens ! -qui refusèrent toute filiation, ces solitaires géniaux de la littérature contemporaine.

  Un Enfant, op.cit., p. 422. ["solche Kreaturen wie du" ; TB, Ein Kind, op.cit., p. 485]. 16 TB, Le Froid, op.cit., p. 349. ["damit du siehst, was du wert bist" ; TB, Die Kälte, op.cit., p. 401]. 17 TB, Un Enfant, op.cit., pp. 405-407. relation, mais Thomas enfant y lit un manque d'amour. La réconciliation n'aura lieu que sur le lit de mort de sa mère.

13

Ibid., p. 349.

14

Ibid., p.348. 15 TB, 18 Ibid., p. 423. 19 Ibid., p. 421. ["Unglück", "Unfriedenstifter", "Lügner" ; TB, Ein Kind, op.cit., p. 483]. 20 Ibid., pp. 485-486. ["Unruhestifter", "Bettnässer" ; Ibid., p. 554]. 21 Ibid., p. 486. ["neuen, beinahe tödlichen Titel" ; Ibid., p. 554]. 22 Ibid., p. 428. 23 Ibid., p. 423.

cette

  32 TB, La Cave, op.cit., p. 129. ["ich bin immer der Störenfried geblieben" ; TB, Der Keller, op.cit., p. 150]. 33 TB, Un Enfant, op.cit., p. 422. 34 Louis Huguet, Thomas Bernhard ou le silence du sphinx, Presses Universitaires de Perpignan, 1991, pp. 16-18.

35

TB, Un Enfant, op.cit., p. 428. 36 Hans Höller, Thomas Bernhard, une vie, Paris, L'Arche, 1994, p. 140. 37 TB, Un Enfant, op.cit., p. 422. 38 TB, Le Froid, op.cit., p. 375. 39 Ibid., p. 350. 40 Louis Huguet, Thomas Bernhard ou le silence du sphinx, op.cit., pp. 165-166.

TB, Un Enfant, op.cit., p. 431.