Sonja K Fagervold
email: fagervold@obs-banyuls.fr

P Lebaron

Evaluation of the degradation capacity of WWTP sludge enrichment cultures towards several organic UV filters and the isolation of octocrylene-degrading microorganisms

Keywords: UV filters, Degradation, Wastewater sludge, Pure bacterial strains 1. Introduction

Organic UV filters are present in wastewater treatment plants (WWTPs) due to the use of these compounds in many personal care products (PCPs) and their subsequent release into the wastewater system from showering/bathing. Once in the wastewater system, organic UV filters generally partition into the solid phase but might also undergo other processes, such as degradation by microorganisms. To further understand the fate of organic UV filters in WWTPs, the degradation of 7 UV filters by WWTP sludge was investigated The UV filters 2-ethylhexyl salicylate (ES), homosalate (HS), butyl methoxydibenzoylmethane (BM) and octocrylene (OC) were degraded after 20-60 days. The rest of the filters tested, namely, bisethylhexyloxyphenol methoxyphenyl triazine (BEMT), methylene bis-benzotriazolyl tetramethylbutylphenol (MBBT) and diethylhexyl butamido triazone (DBT), did not degrade even after 120 days of incubation. The microbial community from the microcosms degrading ES, HS, OC and BM was transferred every 30 days into new microcosms to enrich for microorganisms capable of utilizing the individual UV filters for growth. The enrichment cultures continued to degrade throughout 20 transfers. The microbial community was clearly different between the enrichments degrading ES, HS, OC and BM, meaning that the microbial community was strongly influenced by the UV filter present. Furthermore, several strains were isolated from OC-degrading cultures and two of these strains, Gordonia sp. strain OC_S5 and Sphingopyxis sp. strain OC_4D, degraded OC with and without other carbon sources present. These experiments show that several organic UV filters can be degraded by a specific set of microorganisms. The lack of degradation observed for BEMT, MBBT and DBT is probably due to limited bioavailability. Indeed, this is the first biodegradation study of these filters, in addition to being the first description of ES and HS degradation in microcosms experiments.

UV filters used in personal care products include organic compounds of varying size and characteristics, as well as mineral UV filters, like titanium dioxide (TiO2) and zinc oxide (ZnO). UV filters are important ingredients in sun protection products (e.g., sunscreens), personal care products, plastics, paints and textiles [START_REF] Ramos | Advances in analytical methods and occurrence of organic UV-filters in the environment -A review[END_REF]. Due to their increasing use, environmental contamination by UV filters has become an increasing concern since UV filters are now widely distributed in the environment [START_REF] Sánchez-Quiles | Are sunscreens a new environmental risk associated with coastal tourism?[END_REF][START_REF] Huang | Risks of organic UV filters: a review of environmental and human health concern studies[END_REF]. The main sources of UV filters in the environment can be direct via recreational activities such as bathing but also indirect via wash-off, showering and laundry that end up in wastewater treatment plants (WWTPs). As a consequence, influents from WWTPs contain varying concentrations of UV filters [START_REF] Ramos | A review of organic UV-filters in wastewater treatment plants[END_REF]. For example, BP3, a widely studied organic UV filter, has been found in concentrations over 1,000 ng/L in influents [START_REF] Kasprzyk-Hordern | The removal of pharmaceuticals, personal care products, endocrine disruptors and illicit drugs during wastewater treatment and its impact on the quality of receiving waters[END_REF]Liu et al., 2012a;[START_REF] Biel-Maeso | Removal of personal care products (PCPs) in wastewater and sludge treatment and their occurrence in receiving soils[END_REF] with a maximum measured concentration of 7,800 ng/L [START_REF] Balmer | Occurrence of Some Organic UV Filters in Wastewater, in Surface Waters, and in Fish from Swiss Lakes[END_REF]. To understand the fate of UV filters and their ultimate effect on the environment, it is important to understand the processes UV filters are subjected to in WWTPs. Mineral and organic UV filter undergo completely different processes and while it is important to understand the behavior of both types of UV filters, the focus of this study is on organic UV filters. Generally, WWTPs are not equipped to remove all organic pollutants from the wastewater stream. Thus, incomplete removal of organic UV filters in WWTPs represents a major route of input of organic

UV filters into the environment

The present study focused on a group of organic UV filters with varying molecular weights and solubility, including 2-ethylhexyl salicylate (ES), homosalate (HS), butyl methoxydibenzoylmethane (BM), octocrylene (OC) bis-ethylhexyloxyphenol methoxyphenyl triazine (BEMT), methylene bis-benzotriazolyl tetramethylbutylphenol (MBBT) and diethylhexyl butamido triazone (DBT) (see Table 1). From this list, OC is the UV filter that has been detected at the highest frequency in WWTPs and the highest influent and effluent concentrations (Table 1).

HS concentrations have often been below the detection or quantification limit in WWTP influents and/or effluents, but some concentrations from approximately 600 ng/L to over 1,600 ng/L have been reported in influents [START_REF] Biel-Maeso | Removal of personal care products (PCPs) in wastewater and sludge treatment and their occurrence in receiving soils[END_REF][START_REF] Tsui | Seasonal occurrence, removal efficiencies and preliminary risk assessment of multiple classes of organic UV filters in wastewater treatment plants[END_REF](Table 1). ES has been measured at concentrations of over 1,100 ng/L in influents but is generally present at lower concentrations in effluents.

BM has only been detected in a few studies, with influent concentrations up to 2,935 ng/L, effluent concentrations up to 1,018 ng/L (Rodil et al., 2009a;[START_REF] Tsui | Seasonal occurrence, removal efficiencies and preliminary risk assessment of multiple classes of organic UV filters in wastewater treatment plants[END_REF][START_REF] Cunha | Dispersive liquid-liquid microextraction followed by microwave-assisted silylation and gas chromatography-mass spectrometry analysis for simultaneous trace quantification of bisphenol A and 13 ultraviolet filters in wastewaters[END_REF] and sludge concentrations up to 510 ng/g (dw) (Rodil et al., 2009b). DBT has only been measured and detected in sludge from Belgium at concentrations up to 136 ng/g (dw) (Rodil et al., 2009b), while BEMT and MBBT have never been measured in WWTPs to our knowledge.

The fact that UV filters are found at lower concentrations in effluents than in influents [START_REF] Ramos | A review of organic UV-filters in wastewater treatment plants[END_REF] (Table 1) does not mean that UV filters are degraded. Indeed, once in the WWTP system, the UV filters can either i) stay sorbed to the sludge and thus be removed with the solid phase, ii) stay solubilized and be present in the effluents or iii) be degraded by microorganisms in the WWTP (or the anaerobic sludge digester). Most organic UV filters are hydrophobic substances with log KOW above 4, and thus, their elimination is strongly influenced by sorption processes. Indeed, sorption into solids is often the main factor for the elimination of organic UV filters [START_REF] Kupper | Fate and removal of polycyclic musks, UV filters and biocides during wastewater treatment[END_REF]Liu et al., 2012a). For the complete understanding of the removal processes of organic UV filters in WWTPs, it is necessary to perform mass balance calculations, which can be difficult to achieve. However, this approach has been successful in indicating the degradation of several organic UV filters in WWTPs [START_REF] Biel-Maeso | Removal of personal care products (PCPs) in wastewater and sludge treatment and their occurrence in receiving soils[END_REF][START_REF] Kupper | Fate and removal of polycyclic musks, UV filters and biocides during wastewater treatment[END_REF]Liu et al., 2012a).

An alternative way one can prove that the organic UV filters are indeed degraded is to perform ex situ experiments. The addition of the fungus Trametes versicolor to sludge or slurry has been shown to be beneficial for the degradation of several UV filters [START_REF] Badia-Fabregat | Degradation of UV filters in sewage sludge and 4-MBC in liquid medium by the ligninolytic fungus Trametes versicolor[END_REF][START_REF] Rodríguez-Rodríguez | Removal of pharmaceuticals, polybrominated flame retardants and UVfilters from sludge by the fungus Trametes versicolor in bioslurry reactor[END_REF]Gago-Ferrero et al., 2012). Similar results were found with the addition of a yeast species to sludge [START_REF] Fujii | Degradation of Benzophenone, a Potential Xenoestrogen, by a Yeast Isolated from the Activated Sludge of a Sewage Treatment Plant in Hokkaido[END_REF]. Furthermore, BP3 has been shown to degrade in microcosms of WWTP sludge under different conditions (Liu et al., 2012b). Another important step forward to the understanding of the removal processes is to characterize and identify UV filter-degrading microorganisms. To this end, a Methyliphilus sp. strain FP-6, capable of BP3 degradation, has been identified from WWTP enrichment cultures (Jin et al., 2019). Furthermore, a Mycobacterium agri strain capable of OC degradation has been identified through "reverse discovery" [START_REF] Suleiman | Microbial biofilm formation and degradation of octocrylene, a UV absorber found in sunscreen[END_REF]. However, other carbon sources were available, and thus, the degradation is most likely cometabolic. Finally, our group recently identified a BP3degrading microorganism in WWTP sludge, Sphingomonas wittichii strain BP14P, that degraded BP3 in a growth-dependent manner. The identification and isolation of this strain was made possible by the addition of relatively high concentrations of BP3 to WWTP sludge microcosms followed by multiple serial transfers [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. These factors put pressure on the microbial community to enrich for microorganisms capable of BP3 degradation.

The goal of this study was to investigate i) whether several UV filters with different structures and characteristics could be degraded by WWTP sludge microorganisms, ii) to investigate the microbial community in degrading enrichment cultures, and iii) to isolate degrading microorganisms from at least one of the cultures. To this end, the same process that successfully resulted in the identification and isolation a BP3 degrading microorganisms [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF] was applied to an expanded range of UV filters.

Materials and methods

Chemicals

Several of the UV filters used for biodegradation studies, namely, ES, HS, BM, OC, BEMT and DBT, were provided by Pierre Fabre Pharmaceuticals, while MBBT was purchased from Sigma-Aldrich (Saint-Quentin-Fallavier, France). Analytical standards for the UV filters mentioned above were purchased from Sigma-Aldrich.

Analytical-grade dichloromethane (DCM), 1-methyl-2-pyrrolidinone (NMP), methanol, formic acid (98%) and molecular-grade formaldehyde were acquired from Sigma-Aldrich. Pure water was obtained from an Elga Purelab Flex System (Veolia LabWater STI, Antony, France). Glassware was cleaned with DCM and calcinated at 450 °C for 2 hours to remove all traces of organic matter.

Sampling of sludge and initial culture setup

Sludge sampling and setup of the initial enrichment cultures were performed as described previously [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF] with no modifications except that different UV filters were utilized. Briefly, sludge from Argeles-sur-mer (France) WWTP was collected and used for initial degradation experiments. Sludge (100 mL) was mixed with freshwater mineral media (500 mL) in 1 L glass flasks. UV filters dissolved in methanol were added to the sludge slurries to obtain a final concentration of 10 µg/mL. These live cultures were incubated at 25 °C in the dark in parallel with sterile controls (SC); see [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF] for more details.

Enrichment cultures and biodegradation assays

Enrichment cultures were set up with inoculum from the flasks that showed degradation of different UV filters, namely, OC, ES, HS and BM, as described previously [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. The theoretical UV filter concentrations were 100 µg/mL from the second transfer (Enr 2), and the cultures were incubated in 100 mL Erlenmeyer flasks containing 2 g of inert sand and minimal freshwater media with no other carbon source added except the individual UV filter. The cultures were transferred each month (1% vol/vol).

Biodegradation assays were performed as described previously [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. Briefly, the assays were performed in 15 mL glass tubes with Teflon lined caps (Pyrex, Analytic lab, France) and contained 0.2 g of inert sand, 3 mL of minimal freshwater media and the target UV filter at a concentration of approximately 100 µg/mL. Inoculate for the biodegradation assays were either 150 µL enrichment culture or 150 µL washed cell suspension for the isolates. The tubes were incubated at 25 °C in the dark on a rotary shaker at 100 rpm, and triplicate tubes were sacrificed for extractions at each time point.

For the degradation assay with single strains, the cell numbers were determined by flow cytometry as described previously [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF].

UV filter extractions and HPLC analysis

Two different procedures were used for UV filter extractions, one for the initial biodegradation experiments and another for the biodegradation assays for enrichment cultures and isolated strains, while the HPLC analysis remained the same. These procedures were described previously in detail [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF].

Extraction procedure for the initial degradation experiment and biodegradation assays

To determine the degradation potential in initial WWTP sludge microcosms, the UV filter concentration was determined in a live culture and compared with a sterile control (SC) over time. Briefly, duplicate 5 ml samples were taken from each flask at each time point. The slurry was centrifuged (800 g for 20 min), and the aqueous and solid phases were extracted separately. The extracts were dried and dissolved in NMP/water (8:2, v/v) before HPLC analysis. The recovery rate for the slurries ranged from 50% for ES to 95% for OC (data not shown). For the biodegradation assay, the whole tube containing sand and liquid media was sacrificed for extraction at each time point. Here, the solid and liquid phases were directly extracted in DCM; see [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF] for details. As reported previously, the recovery rate of the different UV filters for the degradation assays ranged from 63 % for ES to 93 % for OC [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF].

HPLC analysis

The UV filters were analyzed using an Ultimate 3000 TM HPLC system equipped with a DAD detector (Thermo Fisher Scientific) and a Phenomenex Kinetex Biphenyl 2.6 µm, 150 x 4.6 mm column as previously described [START_REF] Fagervold | Occurrence and Environmental Distribution of 5 UV Filters During the Summer Season in Different Water Bodies[END_REF][START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. Briefly, the solvent carriers were acidified (0.1% formic acid) water and methanol, starting with a 2:8 ratio of methanol:water, held for 10 minutes, followed by a gradual increase of methanol up to 100%, tailed by 100% methanol for 10 minutes. Oven temperature was 50°C, the flow rate was 1 mL min -1 and the injection volume was 5 µL. The data acquisition software was Chromeleon™ 7.2 (Thermo Fisher Scientific). Calibration curves and retention times of all the different UV filters were determined as described by [START_REF] Fagervold | Occurrence and Environmental Distribution of 5 UV Filters During the Summer Season in Different Water Bodies[END_REF].

Microbial community analysis of enrichment cultures and strain isolation

The microbial community was analyzed by Automated Ribosomal Intergenic Spacer Analysis (ARISA) and Illumina sequencing as previously described [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. Briefly, DNA from enrichment cultures was extracted using a Maxwell® 16 LEV Blood DNA Kit (Promega, Charbonnières-les-Bains, France) with the Maxwell® 16 MDx Instrument (AS3000; Promega). ARISA was performed as described by Fisher et al. [START_REF] Fisher | Automated approach for ribosomal intergenic spacer analysis of microbial diversity and its application to freshwater bacterial communities[END_REF] using the intergenic spacer primers 1406F and 23SRY, with minor modifications as described by [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. For the analysis of the resulting chromatograms, individual fragment lengths (within a 0.1 bp "error range") were considered separate operational taxonomic units (OTUs), and the relative abundance of each OTU was calculated by dividing the area of the peak by the total area of the chromatogram after 200 bp. It is also important to note that only peaks contributing to more than 1% of the relative peak area were taken into account.

For details on the Illumina sequencing and sequence processing, see [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. Sequencing was performed by RTL genomics (Lubbock, Texas, USA) using the bacterial primers 341F (CCTACGGGNGGCWGCAG) and 806R (GACTACHVGGGTATCTAATCC) (Herlemann et al., 2011). The sequences were processed using QIIME 2 (Bolyen et al., 2019). The different samples yielded approximately 27 000 to 56 000 sequences per sample, divided into 244 different OTUs. For further analysis, the focus was on OTUs with a relative abundance over 1% in any of the sequenced samples resulting in 33 OTUs. These OTUs have accession numbers OL468926-OL468958.

OC-degrading strains were isolated on R2B agar, screened and sequenced as described previously [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. The accession numbers for the isolated strains are: OL457616 for strain OC_4D, OL457617 for strain OC_S5, OL457618 for strain OC_S4 and OL457619 for strain OC_S1.

Results and discussion

Degradation of UV filters in initial sludge microcosms

The potential of WWTP sludge to degrade the different UV filters was assessed in a preliminary experiment. The temporal evolution of the UV filter concentration in amended flasks containing active sludge was compared with the concentration of the UV filters in the amended flasks containing sterilized sludge. Degradation was clearly observed for ES, HS, OC and BM after 20-60 days of incubation (Figure 1) in live cultures. However, the concentration seems to increase in some of the sterile control samples, for example for ES and HS. This phenomenon is probably due to a "matrix effect" caused by the sterilization process (autoclaving) changing the extraction efficiency with time. This phenomenon has been observed in other experiments, when autoclaving samples with high organic content (data not shown).

Nevertheless, degradation is evident for ES, HS and BM with a sharp decrease in live cultures over time. Although less evident, OC decreased significantly in the live culture. These initial microcosms were not analyzed further once degradation was observed because they were selected for more detailed experiments described below.

It should be noted that the solid and aqueous phases of the sludge was extracted separately [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF], and that the UV filters partitioned strongly into the solid phase. In fact, the UV filters could barely be detected in the aqueous phase (data not shown). Thus, as previously shown for BP3 [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF], ES, HS, OC and BM are clearly bioavailable for degradation even when partitioned into the solid phase, as the concentration decreased with time in live slurries but not in the sterile controls.

Several of the UV filters did not degrade even after 120 days of incubation, namely, BEMT, MBBT and DBT. These UV filters have log P values over 10 (Table 1), and thus, the lack of degradation in the sludge might be due to strong sorption to the solid phase and hence poor bioavailability for the degrading microorganisms.

Another reason might be the high molecular weight of the target compounds. Indeed, it is thought that molecules have to be less than 600 Daltons to pass through the bacterial cell wall through porin proteins [START_REF] Weiss | Molecular architecture and electrostatic properties of a bacterial porin[END_REF], and BEMT, MBBT and DBT are very large molecules with molecular weights of 600 g/mol. Since no degradation of BEMT, MBBT and DBT was detected in sludge slurries after 120 days, no further enrichments or incubations were initiated using these filters.

Enrichment cultures

Slurry from the flasks that showed biodegradation activity (OC, ES, HS and BM) was transferred to enrich putative UV filter-degrading microorganisms as described previously [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. The enrichments were set up in 100 mL Erlenmeyer flasks that contained 50 mL of minimal media, inert sand, and one of the UV filters. The UV filter concentration was relatively high, with theoretical concentrations of 100 mg/L for the second transfer "enrichment 2". Indeed, this concentration is above the predicted/measured water solubility for OC, HS and BM (see Table 1). It should be noted that the UV filter (BM, HS, ES or OC) was the only available carbon source for the microorganisms in the enrichment cultures. The cultures were transferred every month to further enrich UV filter-degrading microorganisms.

Degradation assays of enrichment cultures

After 5 consecutive transfers, the degradation activity of the enrichment cultures was verified ("Enr 5") by performing biodegradation assays in separate tubes as described in [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. Briefly, biodegradation assays involve the extraction of entire tubes with both the sand phase and the liquid phase. The same procedure was performed on Enr 9 cultures. ES, OC and HS were degraded over 70% in only 5-7 days in Enr 5 and Enr 9 cultures and were approximately 90% degraded after 12-14 days (Figure 2). BM was slower to degrade, with approximately 35% degradation after 7 days in Enr 5 cultures and approximately 50% degradation after 12-14 days. However, with an extended incubation time, the degradation of BM was more complete, with 80% degradation after 28 days in Enr 9 cultures. It was thus clear that the enrichment cultures retained their degradation capability throughout several serial transfers. This degradation activity was observed after more than 20 transfers (data not shown).

Stability of the enrichment culture microbial community through transfers

ARISA was performed to investigate the diversity and stability of the microbial communities in enrichment cultures throughout the transfers. ARISA exploits the fact that different bacteria have different sized intergenic spacers between the 16S rRNA gene and the 23S rRNA gene on their chromosome. Theoretically, one peak in an ARISA profile will correspond to one "OTU", but some bacteria have several copies of the 16S in their genome and give rise to two or more peaks. This analysis was performed on "Day 4" samples of the enrichment cultures degrading ES, HS, BM and OC and after 7, 11, 12 and 20 transfers (Enr 7, Enr 11, Enr 12 and Enr 20).

Generally, 15-28% of the peaks in the enrichment cultures were "prevalent peaks", meaning peaks were detected in all 4 enrichments (Table 2). This means that the microorganism responsible for that peak would be strongly enriched for through 20 transfers over nearly 2 years. Thus, the "prevalent" peaks clearly had a major role in the enrichment cultures. Furthermore, approximately 55-60% of the peaks were found in only one or two of the transfers and generally in smaller amounts. Thus, one can assume that the microorganisms responsible for these peaks do not have a central role in the degradation of the target UV filter.

ES-degrading cultures were the most diverse, containing 29 peaks in total. Eight peaks were prevalent (Figure 3A and Table 2), and the stability of the cultures was relatively high. One prevalent peak stood out as dominant, namely, peak 908, which was present up to 40% in Enr 11. HS-degrading cultures contained 18 peaks, and only 3 peaks were prevalent (Figure 3B). The dominant peak changed with the different transfers; thus, the stability of the microbial community was low. BMdegrading cultures contained 20 peaks, with 5 of these peaks being prevalent (Figure 3C). Although some of the peaks changed with time, the most predominant peak did not seem to change significantly through the different transfers. One can therefore consider the microbial community in BM-degrading microcosms as relatively stable. OC-degrading cultures contained 27 peaks between the four transfers (Figure 3D). Of these peaks, 4 peaks were prevalent. Many of the peaks were present only in small relative abundances, and there was no one dominant peak in any of the enrichments, although several peaks had a relative abundance between 10 and 20%.

Identity of major microorganisms in enrichment cultures

Day 4 samples of Enr 11 and 12 were sequenced to identify the major microbial players in the enrichment cultures (Figure 4). Overall, 33 OTUs with a relative abundance over 1% in any of the sequenced samples were identified. The different samples contained 8-12 OTUs with a relative abundance over 1%, and only one OTU was common in all enrichment cultures, namely, OTU 28, a Betaprotebacteria belonging to the genus Metylophilus. This OTU was only one bp different from Metylophilus luteus Mim T (FJ872109), a restricted facultative methylotroph capable of utilizing both methanol and glucose [START_REF] Gogleva | Methylophilus flavus sp. nov. and Methylophilus luteus sp. nov., aerobic, methylotrophic bacteria associated with plants[END_REF]. However, the exact role of this OTU in the degradation of the various UV filters is not known.

It was evident that the microbial community composition in the different UVdegrading enrichment cultures was very dissimilar (Figure 4). Thus, the presence of an individual UV filter strongly influences the microbial community when transferred multiple times. ES cultures were dominated by two Pseudomonas OTUs within the Gammaproteobacteria, with a total relative abundance of over 70% for the two OTUs. The more abundant of these two OTUs, OTU 33, is one bp different from Pseudomonas benzenivorans strain DSM 8628 (NR_116904). This strain belongs to the Pseudomonas aeruginosa group and is able to grow on benzene, toluene or para-cresol as the sole source of carbon and energy [START_REF] Lang | Pseudomonas benzenivorans sp. nov. and Pseudomonas saponiphila sp. nov., Represented by Xenobiotics Degrading Type Strains[END_REF].

The other Pseudomonas OTU, OTU 32, was only one bp different from several strains, one being Pseudomonas indoloxydans strain IPL-1 (NR_115922), which can oxidize indole [START_REF] Manickam | Description of a novel indoleoxidizing bacterium Pseudomonas indoloxydans sp. nov., isolated from a pesticide-contaminated site[END_REF]. Enrichments degrading the other filters contained fewer Pseudomonas OTUs, and thus, the dominance of Pseudomonas OTUs seemed to be specific for ES-degrading cultures. Indeed, the other enrichments contained more OTUs within the Alphaproteobacteria and/or the Betaprotebacteria.

The most dominant group in the BM-degrading enrichment cultures was Betaprotebacteria, with relative abundances of 55% and 65% in Enr 11 and 12 cultures, respectively. The OTUs within the Betaproteobacteria consisted mainly of 4 OTUs, Comamonas OTU 23, Hydrogenophaga OTU 27, Methylophilus OTU 28 and Thauera OTU 29, and they were present in significant amounts in both the Enr 11 and 12 samples. Methylophilus OTU 28 was described above. The nearest published strain to Comamonas OTU 23, which is 97.8% identical (418 of 427 bp), is Comamonas nitrativorans strain 23310 (NR_025376), a strain that can perform nitrate reduction in the absence of oxygen and can use many different fatty acids as carbon sources but cannot use sugars [START_REF] Etchebehere | Comamonas nitrativorans sp. nov., a novel denitrifier isolated from a denitrifying reactor treating landfill leachate[END_REF]. One can thus envisage that this OTU can be important in the intermediary metabolism of BM and perhaps is not involved in the initial degradation or benzene ring oxidation.

Hydrogenophaga OTU 27 is 100% identical (427 bp) to Hydrogenophaga borbori strain LA-38 (NR_170459). This strain was also isolated from sludge, and contrary to what its name suggests, it cannot grow autotrophically on hydrogen gas but can use various organic molecules for growth [START_REF] Choi | Hydrogenophaga borbori sp. nov., isolated from activated sludge[END_REF]. This OTU seems to be essential for BM degradation, as it is present in over 25% relative abundance in both enrichments sequenced. Thauera OTU 29 is 1 bp different (427 of 428 bp identical) from Thauera humireducens strain SgZ-1 (NR_109534), a facultative anaerobic bacterium isolated from a microbial fuel cell capable of nitrate reduction and the use of various organic molecules as electron donors [START_REF] Yang | Thauera humireducens sp. nov., a humus-reducing bacterium isolated from a microbial fuel cell[END_REF].

The BM-degrading enrichment cultures also contained approximately 20-30% Alphaproteobacteria within the family Sphingomonadaceae. The two main OTUs within this family in BM enrichments were OTU 13 and OTU 15. Their relative % varied between the different samples, but OTU 13 was clearly important in both samples, with relative abundances of over 10% and 16% in Enr 11 and 12, respectively. This OTU was 1 bp different (of 402 bp) from Rhizorhabdus dicambivorans strain Ndbn-20 (NR_151992), a dicamba-degrading strain [START_REF] Yao | Rhizorhabdus dicambivorans sp. nov., a dicamba-degrading bacterium isolated from compost[END_REF], and very similar (400/402 bp identical) to Sphingomonas wittichii RW1 (NR_074268). Indeed, the BP3-degrading activity of Sphingomonas wittichii strain BP14P isolated from BP3 enrichment cultures has recently been described [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. This strain is "OTU 16" here (Figure 4). Thus, it seems that different but very closely related Sphingomonas strains are enriched and are perhaps responsible for the initial degradation of some UV filters.

The microbial community compositions of the two HS samples, Enr 11 and 12, were apparently quite dissimilar in regard to the dominant OTUs present. Although both belonged to the order Sphingomonadales in the Alphaproteobacteria, the dominant OTU in Enr 11 was Sphingopyxis OTU 17 (44% relative abundance), while Erythrobacteraceae OTU 20 dominated in Enr 12, with 50% relative abundance. This kind of "instability" of the microbial community in HS-degrading cultures was also observed in the ARISA experiments (discussed above). Perhaps the ability of HS degradation is less specific and more widespread, meaning that the enzymatic functioning necessary for HS degradation is present in many genera within the Sphingomonadales. OTU 17 was very similar (399/402 bp identical) to several described strains, one of which was Sphingopyxis indica strain DS15 (NR_108185), which was isolated from a hexachlorocyclohexane (HCH)-contaminated site [START_REF] Jindal | Sphingopyxis indica sp. nov., isolated from a high dose point hexachlorocyclohexane (HCH)-contaminated dumpsite[END_REF]. OTU 20 was similar (398/402 bp identical) to several strains within the genera Altererythrobacter/Erythrobacter. There were some differences between the samples from the OC-degrading microcosms but not as much difference as for HS. Some OTUs, such as OTU 1, OTU 24 and OTU 28, were present in both Enr 11 and 12 at approximately the same relative abundance, while other OTUs were present at different ratios in the two samples. For example, OTU 18 (Sphingopyxis sp.) was dominant in the Enr 11 sample, with a relative abundance of 35%. OTU 18 is 100% identical to the described strain Sphingopyxis terrae subsp. ummariensis strain UI2 isolated from a hexachloroclohexane dump site in India [START_REF] Sharma | Sphingopyxis ummariensis sp. nov., isolated from a hexachlorocyclohexane dump site[END_REF]. In Enr 12, another Alphaproteobacterial OTU, OTU11 was most predominant, with almost 40% relative abundance. This was 394/402 bp (98%) identical to a Ferrovibrio xuzhouensis strain isolated from cyhalothrin-contaminated wastewater [START_REF] Song | Ferrovibrio xuzhouensis sp. nov., a cyhalothrin-degrading bacterium isolated from cyhalothrin contaminated wastewater[END_REF].

Perspectives on degradation capacities of enrichment cultures/slurries towards organic UV filters 20

This is the first report of aerobic microcosm degradation experiments for ES, HS BEMT, MBBT and DBT to our knowledge and the first report of ES and HS degradation. OC has been shown to degrade in aquifer materials [START_REF] Liu | Degradation of Six Selected Ultraviolet Filters in Aquifer Materials Under Various Redox Conditions[END_REF] and WWTP microcosms [START_REF] Suleiman | Microbial biofilm formation and degradation of octocrylene, a UV absorber found in sunscreen[END_REF][START_REF] Badia-Fabregat | Degradation of UV filters in sewage sludge and 4-MBC in liquid medium by the ligninolytic fungus Trametes versicolor[END_REF]. Furthermore, there were some indications of OC degradation in marine sediment microcosms [START_REF] Azaroff | Microbial degradation of hydrophobic emerging contaminants from marine sediment slurries (Capbreton Canyon) to pure bacterial strain[END_REF] and in lake water microcosms [START_REF] O'malley | The presence of selected UV filters in a freshwater recreational reservoir and fate in controlled experiments[END_REF]. BM has also been incubated in a sediment/water system, but it was unclear if the removal was due to biodegradation or sorption processes [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]. BM was also tested in lake water microcosms, but no biodegradation was detected [START_REF] O'malley | The presence of selected UV filters in a freshwater recreational reservoir and fate in controlled experiments[END_REF] Moreover, this is the first time that the microbial communities responsible for the degradation of the different UV filters were investigated. Indeed, the fact that the microbial communities were clearly different in the different enrichment cultures suggest that the each different UV filters requires a specific set of microorganisms with a specific set of enzymatic activities for degradation. However, there might be some redundancy in the microbial community since an enrichment of only one specific microorganism was not observed, as seen in BP3 degrading enrichment cultures where one specific strain comprised over 85% relative abundance in enrichment cultures [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF].

Three of the organic UV filters tested, namely, BEMT, MBBT and DBT, clearly did not degrade. The reasons why some of the organic UV filters are degraded while others are not are undetermined. These molecules have different characteristics, such as solubility and size, and thus, the bioavailability may be a determining factor.

Another possibility is that the appropriate microorganisms/enzymes are not present in WWTPs. This might be the case since UV filters apparently require a specific set of microorganism, thus the degradation capability is not universal. One way to determine if this is the case is to screen other environments for degradation activities.

Nevertheless, one might have to come to terms with the idea that some organic UV filters, such as BEMT, MBBT and DBT, are not biodegradable. The question then becomes whether these nonbiodegradable organic UV filters are at all bioavailable to cause toxic effects and/or can be bioaccumulated and biomagnified. Filters such as BP3 and OC have been shown to be bioaccumulative and have been shown to cause toxic effects [START_REF] Peng | Tissue Distribution, Growth Dilution, and Species-Specific Bioaccumulation of Organic Ultraviolet Absorbents in Wildlife Freshwater Fish in the Pearl River Catchment, China[END_REF][START_REF] Lozano | Bioaccumulation and Toxicological Effects of UV-Filters on Marine Species[END_REF], but there are few data to suggest that this is the case for UV filters such as BEMT, MBBT and DBT. These are important factors to keep in mind for the full assessment of risk associated with the use of these organic UV filters.

Isolation of putative degrading microorganisms from OC-degrading cultures

Putative OC-degrading microorganisms were isolated on R2A agar as described previously for BP3-degrading microorganisms [START_REF] Fagervold | Efficient degradation of the organic UV filter benzophenone-3 by Sphingomonas wittichii strain BP14P isolated from WWTP sludge[END_REF]. The focus was on OC because this filter has been detected in WWTP influents, effluents and sludge at relatively high frequencies and at high concentrations [START_REF] Balmer | Occurrence of Some Organic UV Filters in Wastewater, in Surface Waters, and in Fish from Swiss Lakes[END_REF][START_REF] Biel-Maeso | Removal of personal care products (PCPs) in wastewater and sludge treatment and their occurrence in receiving soils[END_REF][START_REF] Kupper | Fate and removal of polycyclic musks, UV filters and biocides during wastewater treatment[END_REF][START_REF] Li | Occurrence and behavior of four of the most used sunscreen UV filters in a wastewater reclamation plant[END_REF]Rodil et al., 2009a;[START_REF] Moeder | At-line microextraction by packed sorbent-gas chromatography-mass spectrometry for the determination of UV filter and polycyclic musk compounds in water samples[END_REF]Liu et al., 2012a;[START_REF] Magi | Emerging pollutants in aquatic environments: monitoring of UV filters in urban wastewater treatment plants[END_REF][START_REF] Langford | Environmental occurrence and risk of organic UV filters and stabilizers in multiple matrices in Norway[END_REF][START_REF] Cunha | Dispersive liquid-liquid microextraction followed by microwave-assisted silylation and gas chromatography-mass spectrometry analysis for simultaneous trace quantification of bisphenol A and 13 ultraviolet filters in wastewaters[END_REF][START_REF] Plagellat | Concentrations and specific loads of UV filters in sewage sludge originating from a monitoring network in Switzerland[END_REF]. Enr 7 cultures were used as inoculum for isolation attempts. After several restreaks on R2A agar, colonies were screened by ARISA to investigate whether the isolates were the same and if they were present in the enrichment cultures. Several strains were successfully isolated from the enrichment cultures, but it should be noted that it was challenging to obtain pure cultures. This difficulty in purifying isolates was previously reported to be a problem [START_REF] Suleiman | Microbial biofilm formation and degradation of octocrylene, a UV absorber found in sunscreen[END_REF]. 11 bacterial types were isolated. Of these 11 initial types, only four isolates could be found in the enrichment cultures (as shown either by ARISA or Illumina sequencing) (see Table 3).

Phylogenetic description of isolated microorganisms

Strain OC_S5 exhibited small orange/pink, dry, well-defined colonies. The 16S rRNA gene sequence was 99.9% identical to Gordonia alkanivorans strain HKI 0136 (NR_026488), a strain isolated from tar-contaminated soil [START_REF] Kummer | Gordonia alkanivorans sp. nov., isolated from tar-contaminated soil[END_REF]. This strain degrades alkenes and is also orange colored. Strain OC_S5 could not be found in the ARISA chromatograms in significant relative amounts but could be detected with a relative abundance of approximately 7% in both Enr11 and 12 in the Illumina sequencing data (OTU 1 in Figure 4). Strain OC_S4 was 99.9% identical to Hydrogenophaga electricum sp. strain AR20 (NR_132676) isolated from an anodic biofilm in a fuel cell [START_REF] Kimura | Hydrogenophaga electricum sp. nov., isolated from anodic biofilms of an acetate-fed microbial fuel cell[END_REF]. This strain can oxidize hydrogen and can also use alternative electron acceptors (nitrate and anode) for anaerobic growth. Hydrogenophaga electricum sp. strain AR20 colonies are glistening and moderate yellow. Isolate OC_S4 was also moderate yellow and glistening with diffuse colonies. Strain OC_S4 was detected in the ARISA chromatograms, and it was shown to be present in approximately 20% relative abundance in the OC-degrading cultures (OTU 24, Figure 4).

Strain OC_4D was over 99% identical to Sphingopyxis terrae subsp. ummariensis strain UI2 (NR_116018), a bacterium isolated from a hexachlorocyclohexane (HCH) dump site [START_REF] Sharma | Sphingopyxis ummariensis sp. nov., isolated from a hexachlorocyclohexane dump site[END_REF]. This strains was metabolically relative versatile, but the authors did not report on the degradation ability of any pollutants [START_REF] Sharma | Sphingopyxis ummariensis sp. nov., isolated from a hexachlorocyclohexane dump site[END_REF]. Strain OC_4D was also yellow, circular with entire margins, as the described strain. This strain is 100% similar to OTU18 that is present in significant amounts in the enrichment cultures (Figure 4 and Table 3).

Strain OC_S1 was 99.4% identical to Pseudomonas delhiensis strain RLD-1 (NR_043731), isolated from a fly ash dumping site in India [START_REF] Prakash | Pseudomonas delhiensis sp. nov., from a fly ash dumping site of a thermal power plant[END_REF].

This strain can use phenantrene as a sole carbon source, but this ability was lost after several streaks on Luria-Bertani (LB) agar. Strain OC_S1 was present in small relative amounts (Table 3) in the OC-degrading enrichment culture.

Interestingly, both Gordonia sp. and Hydrogenophaga sp., in addition to Mycobacterium sp., have been previously reported to be major players in OC-degrading enrichment cultures [START_REF] Suleiman | Microbial biofilm formation and degradation of octocrylene, a UV absorber found in sunscreen[END_REF]. These enrichment cultures also started from WWTP sludge, and the major microbial players were identified by denaturing gradient gel electrophoresis (DGGE) and subsequent sequencing of the major bands in the gel. However, the authors were not able to isolate these strains but used a "reverse discovery approach". This approach involves using deposited strains phylogenetically close to the strains identified by DGGE and sequencing to test their biodegradation ability.

OC degrading activity of isolated strains

The isolated strains were tested for their degradation capacities in minimal media and in media amended with 10% R2B, in addition to surveying the growth by cell counts (Figure 5). The strains were also combined in a mixture to determine any syntrophic effect this would have on biodegradation activity.

Only two of the isolates clearly degraded OC, with or without the presence of other carbon sources. OC_S5 (Gordonia alkanivorans) and OC_4D (Sphingopyxis terrae) degraded approximately 50% of the OC present after only 4 days and over 90 and approximately 80%, respectively, after 20 days. Interestingly, the degradation kinetics seemed to be the same in minimal media and 10% R2B, even though growth was higher in cultures amended with R2B media. Stain OC_S1 (Pseudomonas delhiensis) clearly did not degrade OC and the degradation capacity of strain OC_S4 (Hydrogenophaga electricum) was ambiguous. This strain did not degrade OC in the presence of R2B, but OC seemed to decrease in quantity in some but not all of the replicate cultures after 14 days (Figure 5). Thus, one should consider that strain OC_S4 does not degrade OC until additional verification of these results. Unsurprisingly, clear growth was detected for all the strains when grown on more rich media (minimal media with 10% R2B added). However, some increase in cell numbers in minimal media were also detected for all the strains (Figure 5), even those strains that did not degrade OC. This increase in cell number is probably due to some nutrients being left in the inoculum after the washing procedures. However, this also means that one cannot determine whether the degradation of OC by OC_S5 (Gordonia alkanivorans) and OC_4D (Sphingopyxis terrae) is co-metabolic or growth linked.

These results are in contrast to previous results from [START_REF] Suleiman | Microbial biofilm formation and degradation of octocrylene, a UV absorber found in sunscreen[END_REF], which showed that Gordonia cholesriolivorans did not degrade OC, but Mycobacterium agri showed some degradation activity; however, this was with other carbon sources present. Interestingly, the authors reported biofilm formation around droplets of OC, and the same could be observed here The enrichment culture was not uniform; there were aggregates of bacteria in the media. This observation, together with the fact that we, as Suleiman and coworkers reported [START_REF] Suleiman | Microbial biofilm formation and degradation of octocrylene, a UV absorber found in sunscreen[END_REF], also had difficulties in obtaining pure cultures, suggests that optimal OC degradation is performed by mixed cultures. One can then hypothesize that Sphingopyxis sp. strain OC_4D and perhaps Gordonia sp strain OC_S5 could degrade OC directly while Hydrogenophaga sp. strain OC_S4 and Pseudomonas sp. strain OC_S1, which did not degrade OC directly, have a role in degrading intermediary products in the degradation pathway.

Conclusions

This is the first time the degradation is shown for several of the UV filters (HS and ES) and the first time the microbial community responsible for the degradation is explored with molecular techniques in concurrence with the isolation of OC degrading microorganisms. It is clear that WWTP sludge has the capability to degrade some organic UV filters, such as ES, HS, OC and BM. The degradation activity is stable since the activity can be sustained for over 20 transfers in minimal media. The microbial communities responsible for the degradation of the different UV filters were specific to each UV filter. Thus, the addition of high concentrations of the target UV filter in addition to multiple serial transfers was a successful strategy to enrich specific degrading microbial communities. However, the exact dynamics and the specific role of the major OTUs in the degrading enrichment cultures remain to be determined, except for OC where two strains were confirmed to degrade OC in isolation.

It was also clear that some of the UV filters, namely BEMT, MBBT and DBT did not degrade in these WWTP slurries. The reason for the non-degradation is unclear but is most probably due to poor availability. In this regard one should take into account the seemingly non-bioavailability, in addition to the non-biodegradability of these compounds when performing environmental risk assessments. (Rodil et al., 2009a;Rodil et al., 2009b;[START_REF] Negreira | Sensitive determination of salicylate and benzophenone type UV filters in water samples using solidphase microextraction, derivatization and gas chromatography tandem mass spectrometry[END_REF][START_REF] Negreira | Optimization of pressurized liquid extraction and purification conditions for gas chromatography-mass spectrometry determination of UV filters in sludge[END_REF][START_REF] Kameda | Occurrence and profiles of organic sunblocking agents in surface waters and sediments in Japanese rivers and lakes[END_REF][START_REF] Magi | Emerging pollutants in aquatic environments: monitoring of UV filters in urban wastewater treatment plants[END_REF][START_REF] Wu | Ultrasound-assisted dispersive liquidliquid microextraction plus simultaneous silylation for rapid determination of salicylate and benzophenone-type ultraviolet filters in aqueous samples[END_REF][START_REF] Tsui | Seasonal occurrence, removal efficiencies and preliminary risk assessment of multiple classes of organic UV filters in wastewater treatment plants[END_REF][START_REF] Cunha | Dispersive liquid-liquid microextraction followed by microwave-assisted silylation and gas chromatography-mass spectrometry analysis for simultaneous trace quantification of bisphenol A and 13 ultraviolet filters in wastewaters[END_REF][START_REF] Biel-Maeso | Removal of personal care products (PCPs) in wastewater and sludge treatment and their occurrence in receiving soils[END_REF]

Figures

Figure 1 .Figure 2 .Figure 3 .

 123 Figure 1. Degradation of UV filters in initial sludge cultures. The y-axis represents

Figure 4 .

 4 Figure 4. Bar graph of OTUs present in the different enrichment cultures and their

Figure 5 .

 5 Figure 5. Degradation of OC by different bacterial isolates: strains OC_S1 (A), OC_S4

Table 1 .

 1 Chemical properties and concentrations of organic UV filters used in this study in WWTPs. Note the abbreviations that are used throughout.

	Tables
					WWTP	WWTP	WWTP
					influent	effluent	sludge range
			FW sol.		range (ng/L)	range (ng/L)	(ng/g -dw)
		MW a	(mg/L) b LogP c	[nr. papers] d	[nr. papers] d	[nr. papers] d
	2-Ethylhexyl salicylate (ES) [118-60-5]	250.3	137	5.335	ND -1,188 e [6]	ND -128.9 e [9]	ND -1,370 e [4]
	Homosalate (HS) [118-56-9] 262.3	< LOD (LOD = 0.85)	5.431	ND -1,650.4 f [5]	ND -154.2 f [8]	ND -1,050 f [3]
	Butyl methoxydibenzoylmethane (BM) [70356-09-1]	310.4	19	5.391	250 -2935 g [3]	ND -1,018 g [4]	144 -517 g [1]
	Octocrylene (OC) [6197-30-4]	361.5	9.1	7.083	ND -13,000 h [11]	ND -6,969 h [16]	30-41,610 h [11]
	bis-Ethylhexyloxyphenol
	methoxyphenyl triazine	627.8	0.3	10.627	-	-	-
	(BEMT) [187393-00-6]
	Methylene bis-benzotriazolyl		< LOD
	tetramethylbutylphenol	658.9	(LOD =	15.451	-	-	-
	(MBBT) [103597-45-1]		0.04)
	Diethylhexyl butamido triazone (DBT) [154702-15-5]	766.0	0.2	12.004	-	-	54 -136 i [1]

a MV: molecular weight b Freshwater solubility at 25 °C, experimental data. See protocol in Fagervold et al. (2019). LOD: Limit of detection. c According to Reaxys database (https://www.reaxys.com/, accessed on June 9, 2020) d Number of papers reporting e References for ES concentrations:

Table 2

 2 Number of peaks total in any of the enrichments above 1% relative abundance b Number of peaks present in all 4 enrichments c Number of peaks present in 3 of the 4 enrichments d Peaks only present in one or two of the enrichments Table3. Strains isolated from OC degrading enrichment cultures Identity to the nearest top hit using Basic Local Alignment Search Tool (BLAST) search of describes species (rRNA/ITS databases) b Relative abundance in Enr 7 cultures, this number is used here because this culture was the inoculum for isolation attempts c OTU number from the sequencing analysis and relative percentage in parenthesis d Degradation, verification by in depth biodegradation assay

	. Summary of ARISA results
			ES		HS	BM		OC
			(percentage)	(percentage)	(percentage)	(percentage)
	Nr. of peaks a	29		18	20		27
	Prevalent b	8 (28%)	3 (17%)	5 (25%)	4 (15%)
	Semi prevalent c	3 (10%)	5 (28%)	4 (20%)	7 (26%)
	Spurious d		18 (62%)	10 (56%)	11 (55%)	16 (59%)
							ARISA
	Strain	Closest described strain	Ident. a	fragment	ARISA b Illumina c Deg. d
		(accession number)		length (bp)
	OC_S1 Pseudomonas delhiensis RLD-	99.4 %	768	2.6 %	OTU 31	No
			1 (NR_043731)		(1249/1256)		(1 %)
	OC_S4	Hydrogenophaga electricum	99.9 %	861	5.7 %	OTU 24	?
		sp. (NR_132676)		(1442/1445)		(19%)
	OC_S5 Gordonia alkanivorans strain	99.9 %	700,723,	< 1%	OTU 1	Yes
		HKI 0136 (NR_026488)	(1408/1409)	728,771	(7%)
	OC_4D	Sphingopyxis terrae subsp.	99.9 %	889	6.6 %	OTU 18	Yes
		ummariensis strain UI2 (NR_116018)	(1404/1406)		(25%)
	a

a

f References for HS concentrations:[START_REF] Rodil | Development of a method for the determination of UV filters in water samples using stir bar sorptive extraction and thermal desorption-gas chromatography-mass spectrometry[END_REF]Rodil et al., 2009a;Rodil et al., 2009b;[START_REF] Negreira | Optimization of pressurized liquid extraction and purification conditions for gas chromatography-mass spectrometry determination of UV filters in sludge[END_REF][START_REF] Kameda | Occurrence and profiles of organic sunblocking agents in surface waters and sediments in Japanese rivers and lakes[END_REF][START_REF] Magi | Emerging pollutants in aquatic environments: monitoring of UV filters in urban wastewater treatment plants[END_REF][START_REF] Wu | Ultrasound-assisted dispersive liquidliquid microextraction plus simultaneous silylation for rapid determination of salicylate and benzophenone-type ultraviolet filters in aqueous samples[END_REF][START_REF] Tsui | Seasonal occurrence, removal efficiencies and preliminary risk assessment of multiple classes of organic UV filters in wastewater treatment plants[END_REF][START_REF] Cunha | Dispersive liquid-liquid microextraction followed by microwave-assisted silylation and gas chromatography-mass spectrometry analysis for simultaneous trace quantification of bisphenol A and 13 ultraviolet filters in wastewaters[END_REF][START_REF] Biel-Maeso | Removal of personal care products (PCPs) in wastewater and sludge treatment and their occurrence in receiving soils[END_REF] g References for BM concentrations:[START_REF] Rodil | Development of a method for the determination of UV filters in water samples using stir bar sorptive extraction and thermal desorption-gas chromatography-mass spectrometry[END_REF]Rodil et al., 2009a;Rodil et al., 2009b)[START_REF] Cunha | Dispersive liquid-liquid microextraction followed by microwave-assisted silylation and gas chromatography-mass spectrometry analysis for simultaneous trace quantification of bisphenol A and 13 ultraviolet filters in wastewaters[END_REF] h References for OC concentrations: :[START_REF] Balmer | Occurrence of Some Organic UV Filters in Wastewater, in Surface Waters, and in Fish from Swiss Lakes[END_REF][START_REF] Plagellat | Concentrations and specific loads of UV filters in sewage sludge originating from a monitoring network in Switzerland[END_REF][START_REF] Kupper | Fate and removal of polycyclic musks, UV filters and biocides during wastewater treatment[END_REF][START_REF] Li | Occurrence and behavior of four of the most used sunscreen UV filters in a wastewater reclamation plant[END_REF][START_REF] Rodil | Development of a method for the determination of UV filters in water samples using stir bar sorptive extraction and thermal desorption-gas chromatography-mass spectrometry[END_REF]Rodil et al., 2009a;Rodil et al., 2009b;[START_REF] Pedrouzo | Stir-bar-sorptive extraction and ultra-high-performance liquid chromatography-tandem mass spectrometry for simultaneous analysis of UV filters and antimicrobial agents in water samples[END_REF][START_REF] Moeder | At-line microextraction by packed sorbent-gas chromatography-mass spectrometry for the determination of UV filter and polycyclic musk compounds in water samples[END_REF][START_REF] Negreira | Optimization of pressurized liquid extraction and purification conditions for gas chromatography-mass spectrometry determination of UV filters in sludge[END_REF][START_REF] Liu | Simultaneous determination of benzotriazoles and ultraviolet filters in ground water, effluent and biosolid samples using gas chromatography-tandem mass spectrometry[END_REF][START_REF] Kameda | Occurrence and profiles of organic sunblocking agents in surface waters and sediments in Japanese rivers and lakes[END_REF][START_REF] Gago-Ferrero | Fast pressurized liquid extraction with in-cell purification and analysis by liquid chromatography tandem mass spectrometry for the determination of UV filters and their degradation products in sediments[END_REF][START_REF] Rodríguez-Rodríguez | Removal of pharmaceuticals, polybrominated flame retardants and UVfilters from sludge by the fungus Trametes versicolor in bioslurry reactor[END_REF]Liu et al., 2012a;[START_REF] Rodil | Emerging pollutants in sewage, surface and drinking water in Galicia (NW Spain)[END_REF][START_REF] Badia-Fabregat | Degradation of UV filters in sewage sludge and 4-MBC in liquid medium by the ligninolytic fungus Trametes versicolor[END_REF][START_REF] Magi | Emerging pollutants in aquatic environments: monitoring of UV filters in urban wastewater treatment plants[END_REF][START_REF] Tsui | Seasonal occurrence, removal efficiencies and preliminary risk assessment of multiple classes of organic UV filters in wastewater treatment plants[END_REF][START_REF] Langford | Environmental occurrence and risk of organic UV filters and stabilizers in multiple matrices in Norway[END_REF][START_REF] Cunha | Dispersive liquid-liquid microextraction followed by microwave-assisted silylation and gas chromatography-mass spectrometry analysis for simultaneous trace quantification of bisphenol A and 13 ultraviolet filters in wastewaters[END_REF][START_REF] Biel-Maeso | Removal of personal care products (PCPs) in wastewater and sludge treatment and their occurrence in receiving soils[END_REF] I References for DBT concentrations:(Rodil et al., 2009b)

Acknowledgements:

We would like to thank Cécile Villette for performing the sequencing of the strains and Maeva Duboeuf for help with chemical extractions. This work was carried out in conjunction with the European Marine Biological Resource Centre (EMBRC-ERIC-Banyuls-sur-Mer Oceanographic Observatory, OOB). We are grateful to the BIO2MAR platform for providing access to instrumentation. We would like to thank Christophe Salmeron of the BioPIC Imaging and cytometry platform (OOB) for performing the flow cytometry and the technical support of EMBRC-France, whose French state funds are managed by the ANR within the Investments of the Future program under reference ANR-10-INBS-02. We would also like to thank the local council (Communauté de Communes Albères-Côte Vermeille-Illibéris) who provided access to the WWTP site for sampling. This work was financially supported by Pierre Fabre Dermo-Cosmetic Laboratories in France.


domain: shs scco

text:
PhD Niklas Henke
email: henkeniklas1@googlemail.com

Henke ; Niklas

Design Methods and Innovation Projects as Creativity Consultancy

Keywords: Design, Innovation, Creativity, Consulting, Embodiment

Nowadays design practices can contribute to the competitiveness of companies. The integration of these practices, the methods that characterise them and the imaginary that they convey, represent major evolutions in which innovation agencies play a central role. In this paper, I discuss these design and innovation practices as Creativity Consultancy. These guided practices appear to constitute a specific design and innovation culture marked by the domestication of creativity. The applied rationalization contrasts the practices of designers that are guided by intuition and implicit knowledge. I also describe design and innovation practices based on interviews with designers and innovation project owners, an online survey, and participative observations. Finally, I conclude by locating Creativity Consultancy in the field of tensions between its economic potential and its management challenges before pointing out emerging trends and projecting the identified implications further.

Introduction

Today's design and innovation culture is based on several historic dynamics: The first dynamic is the fusion of information and communication technology (ICT) and cultural industries. The second dynamic is the omnipresence of communicational products in the private, public, and professional realm-a state that is labeled Permanently Online Permanently Connected by [START_REF] Vorderer Peter | Der mediatisierte lebenswandel: Permanently online, permanently connected [the mediatised way of life: Permanently online, permanently connected[END_REF]. The third historic dynamic is the tertiarization of modern, western economies. This tertiarization pushes actors to search for means to differentiate their identities, products, services, and communication. Design and innovation provide the possibility to achieve these objectives by unleashing the potential for competitive differentiation and added semantic value [START_REF] Kogan Anne-France | De quoi la créativité estelle le nom? [what is creativity about?[END_REF]. Thereby, they promote the so-called Attention Economy [START_REF] Franck | Ökonomie der Aufmerksamkeit. Ein Entwurf [The Economy of Attention[END_REF][START_REF] Davenport Thomas | The attention economy[END_REF][START_REF] Citton | L'économie de l'attention: nouvel horizon du capitalisme? [The attention economy[END_REF]. When abundance instead of scarcity becomes the dominating challenge for customers, attention gets a great deal of value which promotes an interest in design and innovation. Design offers the potential to achieve precise and unique identities, differentiate messages, and address affects in a tacit manner [START_REF] Michael | The Tacit Dimension[END_REF](Polanyi, /1966)). Affects are an essential part of communication [START_REF] Fabienne | Anthropologie du corps communicant [anthropology of the communicating body[END_REF]2008;2020). 1 Therefore, addressing the affective dimension of communication in a tacit manner represents a valuable feature for socioeconomic actors [START_REF] Eva | Les sentiments du capitalisme[END_REF][START_REF] Fabienne | Capitalisme affectif: enjeux et pratiques dans les organisations [Affective capitalism: issues and practices in organizations[END_REF]. 1 Commonly, affects are divided into moods, feelings, and emotions and defined according to their valence, intensity, and duration [START_REF] Paul | Wie Sie Emotionen erkennen und richtig interpretieren [Reading emotions. How to recognise and correctly interpret emotions[END_REF]. Their analyses may focus on their biopsychological, interpersonal, or societal dimensions [START_REF] Tcherkassof | Les émotions et leurs expressions [Emotions and their expressions[END_REF]. However, since the goal of this article is to take the totality of affective experiences into account, it explores affects instead of moods, feelings, or emotions. Nevertheless, during the interviews, I used the term emotions instead of affects because it is the common term for designers. Especially, important to this research is the social and corporal dimension of affects [START_REF] Fabienne | Penser le corps affectif comme UN média dans une perspective d'anthropologie par la communication [thinking of the affective body as a medium in a perspective of anthropology through communication[END_REF]2020). I understand affects as grounded in the human body [START_REF] Niedenthal Paula | Embodying emotion[END_REF]. The "corporality of affects" (Henke, 2021) can be divided into an exteroceptive (e.g., smell), proprioceptive (e.g., muscular tensions), and interoceptive (e.g., headache) dimension (Martin-Juchat, 2002, p. 9). Because of this theoretical framework, I asked not only about affects but about the human body in the interviews.

Design and innovation 2 are key in the continuous development of products and services, either by inventing new ones 3 or by improving existing ones. By that, design and innovation are fundamental for the so-called "Creative Destruction" [START_REF] Schumpeter | Capitalism, socialism and[END_REF](Schumpeter, /1942, p. 81), p. 81). They are an important anthropological feature (Goffman, 2014(Goffman, /1956), but the extension of their socioeconomic importance represents a recent phenomenon (Reckwitz, 2019). The changing mindset that comes along with this phenomenon implies a rupture with past traditions and animistic beliefs as well as magical or spiritual heritages, favoring an orientation towards the future [START_REF] Thierry | Innovation et histoire. Une critique philosophique[END_REF]2021). The origins of design and its project mode as we know it today can be traced to the Renaissance [START_REF] Vial Stéphane | De la spécificité du projet en design: Une démonstration [the specificity of the design project: A demonstration[END_REF]. Notably, during the industrialization era, design gained importance and emerged as a profession, an educational branch, and a research 2 I follow the tri-dimensional design definition proposed by Zacklad (2017, p. 5) who defines design according to 1) the dimension of the artifact (form, function, experience, and symbolic), 2) the modalities of engagement (classic project mode, design thinking, participative design, and rhetoric design) and 3) the type of artifact (object, visual, interaction, service, and space). The advantage of this approach is that it allows to precisely define a wide range of design types. My comprehension of innovation follows the diffusion theory of [START_REF] Rogers Everett | Diffusion of Innovations[END_REF]Rogers ( /1962)). According to [START_REF] Rogers Everett | Diffusion of Innovations[END_REF]Rogers ( /1962, p. 15), p. 15), the diffusion of an innovation depends on its relative advantage (the higher the perceived advantage is, the more rapidly the innovation gets diffused), its compatibility (the degree to which the innovation is compatible with the users existing values, experiences, needs, and practices), its complexity (is the innovation difficult to understand?), its testability (is the innovation testable on a limited base?), and its observability (the more visible the effects of the innovation are to others, the more rapid its diffusion is). This approach shows how intimately innovation and design are linked. Both are directed towards the future and meant to improve things [START_REF] Thierry | Innovations: Une enquête philosophique [Innovations: A philosophical enquiry[END_REF]. Therefore, they imply an underlying value set for which they can be criticized [START_REF] Adorno Theodor | Ästhetische Theorie [Aesthetic theory[END_REF](Adorno, /1973)). Furthermore, they share the necessity of creativity. It is important to differentiate between a creative person, product, and process [START_REF] Funke | Zur psychologie der kreativität [on the psychology of creativity[END_REF]. The design and innovation practices that are described in this article englobe all of them. A creative process can be separated into 1) preparation, 2) incubation, 3) comprehension, 4) evaluation, and 5) application (idem). These steps correspond to the maturity stages of an innovation project (Ixiade, 2021a): explore, imagine, experiment, and develop.

3 Keeping aside the question to what extent the creation of something new is possible or to which degree it represents the reassembling of existing elements-as Adorno (1992/1973, p. 36) claimed. Following the definition of [START_REF] Rogers Everett | Diffusion of Innovations[END_REF]Rogers ( /1962)), an innovation is characterized by its perceived novelty and therefore subjective.

discipline [START_REF] Petit | Perspectives sur le design. Métier, enseignement, recherche[END_REF]. The latter one has to be divided into research via, for, or about design [START_REF] Joost Gesche | Design as research. Positions, arguments, perspectives[END_REF]. Design can either be a research method (research via design), a served research goal (research for design), or a research object (research about design). In this article, I focus on design as a research object.

Nowadays, design and innovation penetrate all spheres of society (Dunne & Raby, 2013;[START_REF] Werner | Innovation society today. Perspectives, fields, and cases[END_REF]. Innovation is not limited to the technoscientific domain anymore, which nevertheless represents the most dominant driver of economic growth and competitive advantages [START_REF] Michael | The Most Innovative Companies. The Serial Innovation Imperative[END_REF]. The innovations of the 20 th century (notably innovations in ICT) brought design and innovation closely together. They implicated a shifting focus towards the visual sense, which promoted design professions. For example, in the field of human-machine-interaction, the transition towards graphical user interfaces not only promoted the democratization of computers but also increased the need for user experience (UX) and user interface (UI) designers [START_REF] Paul | Where the action is. The Foundations of Embodied Interaction[END_REF]. Today, it is hard to find an innovation project without participating internal or external designers. Thereby, design professions become interconnected with other communication professions (Miège, 2017), resulting in a new form of design and innovation cultures.

The described tendencies indicate the rising importance of design and innovation, which pushes socioeconomic actors to search for elements that impact the success of an innovation. Having a clearer picture of the success chances of a project and reducing its risk of failure by modifying the innovation concept are the value proposition of innovation agencies. Recently, more attention is paid to the anthropological complexity of users and especially to their affects. Design and innovation practices become less techno-deterministic and more user-centered forming the so-called user-centered turn (Norman, 2013(Norman, /1988). 4 Design influences the perception and comprehension of SFSICinnovation as well as the affective reactions towards it. It is recognized for its potential to address the ontological, pragmatic, and prescriptive dimensions of communication. These dimensions are crucial for the diffusion of an innovation. To take design and affects into account, innovation agencies possess a diversified portfolio of methods that allows them to respond to client demands. 5 Currently, we witness a variety of design and collaboration methods: Most commonly, design thinking is applied and promoted by Stanford, IDEO, the Hasso Plattner Institute, and the British Design Council [START_REF] Kelley Tom | The art of innovation. Lessons in creativity from IDEO america's leading design firm[END_REF][START_REF] Brown Tim | Change by design. How design thinking transforms organizations and inspires innovation[END_REF]. These methods are applied by different professions, and their specific applications often change with each project and the involved persons. They aim to reduce the risk of failure and assure project outcomes while maintaining a sufficient level of efficiency. In the case of the Grenoble-based innovation agency Ixiade, the available methods address each potential source of failure within the innovation process (Ixiade, 2021b). 6 Being able to explicitly address these different sources of failures is an asset regarding the fact that between 80% and 95% of innovation projects fail. 7In this section, I described the historic dynamics that have promoted design and innovation. Thus, it is essential to indicate current design and innovation practices. Based on these descriptions, I locate design methods and innovation agencies in a new form of design and innovation culture that is understood as Creativity Consultancy (Hypothesis 1). The goal of Creativity Consultancy is to operationalize creativity and guide collaboration between team members in an efficient manner. Such a task includes tensions between its socioeconomic potential and its management challenges. The potential lies in the exploitation of creative forces and the challenges facing the integration of their epistemological and sociopsychological features in industrial contexts. To enlighten this field of tensions, I shed light on current design and innovation practices.

Mixed-Method Approach: Interviews, Online Survey, and Participative Observations

The collected data can be divided into two categories: One represents a focus on design practices, whereas the other one focuses on innovation projects. Data about design practices was collected via semi-structured interviews with designers (N = 21), 8 via an online survey with designers (N = 22), 9 and from my own design practice as a UI designer at Ixiade. The data about innovation projects was based on semi-structured interviews with innovation project owners (N = 8)10 as well as participative observations of innovation projects with Ixiade. 11 To gain a broad picture of design and innovation practices, I addressed a diverse range of profiles. I asked about daily working routines, the evaluation of design quality, applied methods, the role of creativity, the perceived link between form and function, the characteristics of ICT for design practices, and the role of affects and the human body. During the interviews, I put a focus on identifying elements that influence design outcomes, such as sources of inspiration and material constraints. When addressing the project owners, I focused on design management and collaboration tools. Based on this mixed methodology, I was able to gain a detailed picture of the design and innovation practices presented below.

Results

The Heterogeneity of Design Practices

The findings show a multitude of design practices. Designers working attitudes ranged from auto-centered to user-centered ones (described precisely in the latter section) and differ by the designer's motivations and aspirations. Most designers work in interdisciplinary teams characterized by a constant exchange with multiple professions-most notably other communication professions. The identified sources of inspiration ranged from project characteristics, its technological and historical context, meetings with the client to daily news, and society as a whole. The elements that constitute design outcomes were the project characteristics (including budgetary, material and temporal constraints, and its context), the dynamics of the teams involved, the practitioner's personality, and randomness.

The Presence (and Absence) of Design Methods and Theory

The results show that designers tended to perceive structured workflows, design methods, and scientific theory as detrimental to creativity and flow. The application of design methods appeared to be mostly selective and higher for project owners than for designers. The application of methods ranged from never for some designers to selective for the majority. I could not identify designers that constantly and explicitly applied methods. Instead, practices were guided by intuition, experience, and implicit knowledge. Certain methods were shared between designers and project owners (e.g., design thinking or creative problem solving [CPS]), but their presence was more dominant for project owners. During innovation projects, I observed a multitude of methods to operationalize creativity, design, and the user. Mastering these methods was perceived as an important value. Creativity was rationalized when creating or modifying an innovation concept in a similar way that users' affects were rationalized to calculate the acceptability of an innovation. None of the surveyed designers stated a notable impact of scientific theories (e.g., perception theory) on their design practices. 12 I thus highlight an absence, an ignorance, or even an aversion towards scientific theories which were perceived as detrimental to creativity.

The Perceived Role of Embodiment

In the survey, the designers associated affects and design closely, while the affective dimension of a design was even taken as an indicator for its quality. This is especially interesting, since defining design quality has become a difficult task for many designers, often underlining its subjective nature. In some cases, design and affects were even viewed as tautological. The perceived importance of affects stands in contrast to the absence of methods that address affects. Only few designers stated to use methods that explicitly take affects into account. More often, affects were considered in an implicit way via qualitative user research methods (mostly via interviews). Moreover, affects played a role as an implicit background disposition between the designer and design. A common attitude was to work until the results feel right. Asked about the role of the human body, its importance was put into the context of the use of the five human senses. It was seen as a mediator between the designer or user and design. In the case of product design, interior design, and stage design, the human body played the most dominant role, especially via ergonomics. In the case of brand design, the human body gained importance via multisensorial brand experiences. Nevertheless, some designers were rather puzzled by the question about the role of the human body in design practices, especially those designers working mainly on a visual level.

From User-Centered to Auto-Centered Working Attitudes

The identified working attitudes ranged from user-centered to auto-centered ones. In the case of user-centered working attitudes, designers met users via interviews, focus groups, or shadowing and attributed high importance to this. In these cases, the value of the work was often perceived in the user feedback. In the case of auto-centered working attitudes, designers considered themselves as archetypical users whose perception and evaluation of a design were seen as representative of the target group. The auto-centered working attitudes approached artistic ones or what Zacklad (2017) defines as "design d'auteur". 13 The user-centered design approaches overlap with other communication professions-notably with marketing, public relations, and strategy. Several moderate approaches could be observed. In these cases, the designer oscillates between auto-and user-centered approaches according to the necessities of each project. In innovation agencies, I observed a similar diversity of approaches. Some agencies construct their offer around user-centered approaches such as the case of Ixiade (2021a) that places the "user experience at the heart of innovation." Others replace the user feedback via their internal creativity practices.

Discussion

The Embodiment of Creativity

Following an embodiment perspective [START_REF] Niedenthal Paula | Embodying emotion[END_REF], creativity can be understood as grounded in the physiological structure of the practitioner. My practice as a UI designer confirms design as an affective and corporal competence. Designers' competence is not only constituted by their aesthetical or technical skills but also by their abilities to integrate themselves into team dynamics or the sensing of non-expressed team preferences or client intentions. Nevertheless, this is not shared by most designers. From this, I conclude that the theoretical framework of embodiment is not an explicit but implicit reality of design and innovation practices. 14 This conceptualization is 13 The term can be translated as signature designer. 14 The role of intuition and implicit knowledge represents a parallel between design practices and scientific production. According to [START_REF] Michael | The Tacit Dimension[END_REF]Polanyi ( /1966)), scientific research needs the sensing of promising problems and directions; without that, they could be justified at that SFSICquite close to the "reflective practitioner" of [START_REF] Schön Donald | The reflective practitioner: how professionals think in action[END_REF], who strengthens the role of spontaneity and intuition in professional practices-aspects that represent corporal and affective competencies. I want to take this further by understanding creativity practices as grounded in the Dasein [START_REF] Heidegger Martin | Sein und Zeit [Being and time[END_REF](Heidegger, /1927) ) of the practitioner, including its sociopsychological, physiological, affective, and temporal dimensions [START_REF] Henke Niklas | La corporéité des affects selon le design dans les projets d'innovation: approche critique, analyse des pratiques, et perspectives [The corporality of affects according to design in innovation projects: critical approach, analysis of practices, SFSICand perspectives[END_REF][START_REF] Henke Niklas | The design turn for the management of public relations: Emerging challenges for communication professionals[END_REF]. This understanding means that a designer is employed for his/her overall Dasein. I combine this with the approaches of [START_REF] Habermas Jürgen | Technik und Wissenschaft als "Ideologie[END_REF] and [START_REF] Herbert | One-dimensional man[END_REF]Marcuse ( /1964) ) who described the inscription of ideology and political power in technology. Transferring this line of thought on the described practices, I conclude that Creativity Consultancy represents an operationalization of the practitioner's Dasein. The Dasein of a designer is operationalized according to the underlying values of the new design and innovation culture.

Implications of the Project Mode

The project mode constitutes the temporal setting of Creativity Consultancy [START_REF] Vial Stéphane | De la spécificité du projet en design: Une démonstration [the specificity of the design project: A demonstration[END_REF]. This phenomenon can be understood in the context of the acceleration of modernity as described by [START_REF] Rosa Hartmut | Die Veränderung der Zeitstrukturen in der Moderne [Acceleration. The change in time structures in modernity[END_REF]2013). The acceleration of creativity adds an additional form of acceleration to the three forms described by Rosa (2013): 1) technological acceleration, 2) acceleration of social change, and 3) acceleration of the pace of life. They are reflected in the acceleration of new product development processes. The project mode can be interpreted as fragmentation of creativity practices similar to the fragmentation of daily life described as acceleration effects by Han (2013) and Rosa ( 2013). Furthermore, it can be linked to the modern tendency towards ahistoric worldviews [START_REF] Baumann Zygmunt | Flüchtige Zeiten: Leben in der Ungewissheit [Liquid Times: Living in an Age of Uncertainty[END_REF]. Ahistoric worldviews promote creativity since identities and meaning must be reconstructed individually and constantly instead of being inherited moment (pp. 21-22). In the same way, design and innovation projects need certain sensing of promising directions that often get justified retrospectively. and remaining fixed. This includes an increasing responsibility of Creativity Consultancy because if the designs or innovations are not satisfying, only Creativity Consultancy is to be held responsible.

Distributed Creativity Practices

In the new design and innovation culture of Creativity Consultancy, design and innovation are realized in interdisciplinary teams with a variety of actors participating in the processes. They contrast the aged idea of a single genius struck by an enlightened thought. Instead of this, creativity evolves distributed between a variety of actors. I propose to label this Distributed Creativitysimilar to the concept of Distributed Cognition [START_REF] Hutchins Edwin | Cognition in the wild[END_REF]. The idea of [START_REF] Hutchins Edwin | Cognition in the wild[END_REF] states that the socially distributed cognition creates a new form of cognition that transcends the one of a single person. Similarly, the distributed creativity practices create a creative competence that transcends the individual ones. Innovation agencies use this to justify the implication of the user. They see the collective creativity as superior to the individual one; therefore, it implies a variety of persons that collaborate via design methods. From an actornetwork perspective [START_REF] Latour Bruno | Reassembling the social: An introduction to actornetwork theory[END_REF][START_REF] Mcmaster Tom | The agency of hybrids: Overcoming the symmetrophobic block[END_REF][START_REF] Bencherki | Action and agency[END_REF], I include the involved materialities as participating actors. The importance of implicated materials and tools during design processes on project outcomes is shown by [START_REF] Smith Maarten | Scaffolding shared imagination with tangible design[END_REF]. This means that design and innovation are constituted by a tacit dialogue between the involved participants and the implicated materialities. Since the user-centered turn, the creation of designs and innovations is partly realized by the user. 15 This represents an externalization of creativity and operationalization of the user. It raises the question of whether the user-centered turn is a euphemism and whether Creativity Consultancy proposes a modern justification of industrial processes.

Creativity Consultancy

I understand the application of design methods and the activity of innovation agencies as a new form of design and innovation culture labeled Creativity Consultancy. Creativity Consultancy guides collaborations between persons in a creative process to assure the efficient realization of design and innovation. Creative forces are accepted for their economic potential but implicate management challenges via their epistemological and sociopsychological features [START_REF] Cross | CSÍKSZENTMIHÁLYI Mihaly, Flow: The psychology of optimal experience[END_REF][START_REF] Gentes | The indiscipline of design[END_REF]. Creativity Consultancy responds to challenges. Nevertheless, there lies a certain aspect of randomness in creation processes, which is necessarily tolerated. Kogan and Andonova (2019, p. 2) describe this aspect as normalization of incertitude. Creativity Consultancy tries to reduce this uncertainty via the rationalization of practices. Therefore, I locate design and innovation in a field of tension between their economic potential on the one hand and their epistemological and sociopsychological management challenges on the other hand.

Conclusion

In this paper, I present design and innovation in their historic context and outline that design methods and innovation agencies can be located in a broader phenomenon that I label Creativity Consultancy (Hypothesis 1). I also describe the essential features of this new form of design and innovation culture. It operationalizes creativity, guides collaborations, and assures outcomes. I finally locate it in the field of tensions between its economic potential and its management challenges. To verify my assumption, I analysed design and innovation practices via semi-structured interviews, an online survey, and participative observations. My findings show the heterogeneity of practices: designs and innovation are constituted by the project characteristics (the material, budgetary, and temporal constraints), the sociopsychological dynamics of the involved teams, the Dasein [START_REF] Heidegger Martin | Sein und Zeit [Being and time[END_REF](Heidegger, /1927) ) of the practitioners, and randomness. Creativity Consultancy aims to reduce the aspect of randomness and the risk of failure, which represents its value proposition. In doing so, creativity becomes rationalized. This rationalization stands in contrast to the practices of designers that are mostly guided by intuition, experience, and tacit knowledge. In this paper, the application of design methods was more dominant for innovation project owners than for designers. The designers in the survey often perceived methods, scientific theory, and structured workflows as detrimental to creativity and flow. Methods were mostly applied selectively. Design and affects were perceived as linked and nearly tautological. Therefore, I hypothesize to explore explicit methods to assure the impact of affects on design practices. This is not the case. Affects play not an explicit but an implicit role as a constant background disposition, especially important for auto-centered working attitudes. I identify creativity practices between auto-centered and user-centered working perspectives. Auto-centered attitudes take the practitioner as an archetypical user and his/her perception as representative for the target group. Whereas user-centered attitudes focus on the integration of the user and perceive an added value in this integration. I interpret the integration of the user as an externalization of creativity that I describe as Distributed Creativity. The paradigm Be creative! And innovate! represents a paradox and at the same time an economic obligation that Creativity Consultancy valorizes.

In this article, I demonstrate how design methods and the activity of innovation agencies can be understood as Creativity Consultancy. I identify their practices as a tacit dialogue between the project characteristics, the involved practitioners (including the user), and the implicated materialities. This represents a rising convergence between the social and the economic sphere. Thereby, design methods and innovation agencies deliver a modern argument to justify industrial processes, creating a new design and innovation culture.

This is, for example, expressed in the CAUTIC © method (Mallein & Toussaint, 1994) that takes sociopsychological and ethical dimensions into account.

For a classification of the strategic positioning of innovation agencies, seeGlennie and Bound (2016, pp. 22-25). As an example of the method portfolio of an innovation agency, seeIxiade (2021b, pp. 26-35).

The identified sources of failure are insufficient product definition, insufficient knowledge about the target group, insufficient knowledge about the addressed market segments, wrong timing, disproportionate estimation of the market value, insufficient consideration of the context, targeting errors, missing innovation culture, missing methods, and psychological innovation bias(Ixiade, 2021b).

For example,

80% were mentioned by UMI (2020) and 95% were noted by Nobel (2011). The differences in failure rates depend on the maturity of the project at which one begins the calculation. According toStevens and Burley (1997, p. 7), the gap is even more striking, claiming that 99.7% of submitted ideas and 40% of launched projects fail.

Just to mention a few examples, their official profiles were communication designer, web designer, product designer, UI designer, UX designer, stage designer, animation designer, creative director, eco-designer, head of color and trim design, professor of interior design, professor of corporate identity.

The online survey represented a summary of the interview guide. The interview guide was synthesized into eight key questions: 1) What are your spontaneous associations towards design and emotion? 2) How do you judge design quality? 3) What are the most influential elements for your daily practices? 4) Which methods do you apply during your daily practices? 5) How do you describe the relation between form and function? 6) Which role plays the human body for your daily practices? 7) How do you describe the interaction between design, emotions and the human body? 8) The questionnaire was closed with several sociodemographic questions (e.g., age and education) and questions about the professional context (e.g., design types and size of the agency). The goal of this online survey was to complement the designers' oral expressions (interviews) with textual ones.

The consulted profiles were head of innovation management, design manager, scientific management advisor, innovation project manager, marketing manager, innovation consultant, chief experience officer (CXO), and market researcher.

The data was collected during a Ph.D. thesis with Ixiade between 2017 and 2020. Therefore, the observations are limited to one agency. Conducting observations with other agencies could strengthen the dataset. A limitation to this agency was necessary because of the CIFRE convention via which the Ph.D. thesis was financed. The observations are based on my experience as a UI designer during these three years, witnessing around 30 innovation projects. The projects showed different states of maturity: from vague ideas via precise concepts and prototypes towards ergonomic tests of already existing products and interfaces.

I did not present any perception theory but explored existing knowledge about scientific theories that may play a role in design practices according to practitioners. Some of the few mentioned approaches were the archetypes of C.G. Jung or the "flow" concept fromCsíkszentmihályi (1990).

The integration of the user was more present for the participative observation and for the interviewed project owners than for the design practices described by the interrogated designers. The minority of designers showed user-centered working attitudes and the majority showed auto-centered working attitudes.

Acknowledgments

I acknowledge Pascal Pizelle and Isabelle Fournié for the realization of the CIFRE convention that served this research and Fabienne Martin-Juchat for her comments while writing this article.