Examining functional group-dependent effects on the ionization of lignin monomers using supercritical fluid chromatography/electrospray ionization mass spectrometry.

The chemical and biological conversion of biomass-derived lignin is a promising pathway for producing valuable low molecular weight aromatic chemicals, such as vanillin or guaiacol, known as lignin monomers (LMs). Various methods employing chromatography and electrospray ionization-mass spectrometry (ESI-MS) have been developed for LM analysis, but the impact of LM chemical properties on analytical performance remains unclear. This study systematically optimized ESI efficiency for 24 selected LMs, categorized by functionality. Fractional factorial designs were employed for each LM to assess ESI parameter effects on ionization efficiency using ultra-high-performance supercritical fluid chromatography/ESI-MS (UHPSFC/ESI-MS). Molecular descriptors were also investigated to explain variations in ESI parameter responses and chromatographic retention among the LMs. Structural differences among LMs led to complex optimal ESI settings. Notably, LMs with two methoxy groups benefited from higher gas and sheath gas temperatures, likely due to their lower log P and higher desolvation energy requirements. Similarly, vinyl acids and ketones showed advantages at elevated gas temperatures. The retention in UHPSFC using a diol stationary phase was correlated with the number of hydrogen bond donors. In summary, this study elucidates structural features influencing chromatographic retention and ESI efficiency in LMs. The findings can aid in developing analytical methods for specific technical lignins. However, the absence of an adequate number of LM standards limits the prediction of LM structures solely based on ESI performance data.

Priority and emerging organic microcontaminants in three Mediterranean river basins: Occurrence, spatial distribution, and identification of river basin specific pollutants.

There is a worldwide growing use of chemicals by our developed, industrialized, and technological society. More than 100,000 chemical substances are thus commonly used both by industry and households. Depending on the amount produced, physical-chemical properties, and mode of use, many of them may reach the environment and, notably, the aquatic receiving systems. This may result in undesirable and harmful side-effects on both the human and the ecosystem's health. Mediterranean rivers are largely different from Northern and Central European rivers in terms of hydrological regime, climate conditions (e.g. air temperature, solar irradiation, precipitation), and socio-economics (e.g. land use, tourism, crop types, etc.), with all these factors leading to differences in the relative importance of the environmental stressors, in the classes and levels of the pollutants found and their environmental fate. Furthermore, water scarcity might be critical in affecting water pollution because of the lowered dilution capacity of chemicals. This work provides raw chemical data from different families of microcontaminants identified in three selected Mediterranean rivers (the Sava, Evrotas, and Adige) collected during two sampling campaigns conducted in 2014 and 2015 in three different matrices, namely, water, sediments, and biota (fish). More than 200 organic micropollutants were analyzed, including relevant groups like pharmaceuticals, personal care products, perfluorinated compounds, pesticides, pyrethroid insecticides, flame retardants, and persistent organic pollutants. Data obtained were summarized with some basic statistics for all compound families and matrices analyzed. Observed occurrence and spatial patterns were interpreted both in terms of compound physical-chemical properties and local environmental pressures. Finally, their spatial distribution was examined and their ecotoxicological risk in the water phase was assessed. This allowed locating, at each basin, the most polluted sites ("hot spots") and identifying the respective river basin specific pollutants (RBSPs), prioritizing them in terms of the potential ecotoxicological risk posed to the aquatic ecosystems.

Anthropogenic contaminants in freshwater from the northern Antarctic Peninsula region.

This study aimed to evaluate the presence of ultraviolet filters (UV-Fs), benzotriazoles, pyrethroids and per- and polyfluoroalkyl substances (PFASs) in freshwater and wastewater from the northern Antarctic Peninsula region. All water samples analyzed contained UV-Fs residues and high concentrations were detected in anthropogenic impacted sites (< LOD up to 1300 ng/L). Likewise, benzotriazoles were detected in all water samples (< LOQ-920 ng/L). Regarding suspended particulate matter, almost all UV-Fs and all benzotriazoles were measured at concentrations ranging from < LOQ to 33 µg/g dry weight. Pyrethroids were also detected (< LOQ-250 ng/L) and their presence implies the existence of a gateway to the Antarctica Peninsula from other regions. The data confirmed the presence of PFASs (< LOD-7500 ng/L) in this area, in agreement with previous studies. In light of these results, extended monitoring in Antarctica should be carried out to perform a reliable environmental risk assessment leading to propose recommendations to minimize the anthropic impact.

Engineering the carotenoid biosynthetic pathway in Rhodothermus marinus for lycopene production.

Rhodothermus marinus has the potential to be well suited for biorefineries, as an aerobic thermophile that produces thermostable enzymes and is able to utilize polysaccharides from different 2nd and 3rd generation biomass. The bacterium produces valuable chemicals such as carotenoids. However, the native carotenoids are not established for industrial production and R. marinus needs to be genetically modified to produce higher value carotenoids. Here we genetically modified the carotenoid biosynthetic gene cluster resulting in three different mutants, most importantly the lycopene producing mutant TK-3 (ΔtrpBΔpurAΔcruFcrtB::trpBcrtB T.thermophilus ). The genetic modifications and subsequent structural analysis of carotenoids helped clarify the carotenoid biosynthetic pathway in R. marinus. The nucleotide sequences encoding the enzymes phytoene synthase (CrtB) and the previously unidentified 1',2'-hydratase (CruF) were found fused together and encoded by a single gene in R. marinus. Deleting only the cruF part of the gene did not result in an active CrtB enzyme. However, by deleting the entire gene and inserting the crtB gene from Thermus thermophilus, a mutant strain was obtained, producing lycopene as the sole carotenoid. The lycopene produced by TK-3 was quantified as 0.49 â€‹g/kg CDW (cell dry weight).

Nanosized titanium dioxide UV filter increases mixture toxicity when combined with parabens.

To address the concern about the environmental impact of engineered nanoparticles frequently used in the recently marketed personal care and hygiene products (PCPs), we conducted a toxicity assessment and determined the EC50 values of the nanosized inorganic UV filter TiO2 (nano-TiO2), as well as those of the organic UV filter oxybenzone (BP3) and three parabens (methyl, propyl, and benzylparaben) present in most PCPs formulation. The bioassays were carried out through standardized toxicity bioassays on two environmentally relevant aquatic species i.e. Daphnia magna and Phaeodactylum tricornutum. For nano-TiO2 48 h EC50 on D. magna was 3.09 mgL-1 and for parabens ranged from 32.52 to 1.35 mgL-1. The two most toxic compounds on D. magna, nano-TiO2 and benzylparaben (BzP), were further tested with the algae. For nano-TiO2 72 h EC50 value was 2.27 mgL-1 and for BzP it was 10.61 mgL-1. In addition, D. magna was exposed to selected binary mixtures of the target compounds i.e. nano-TiO2+BP3, nano-TiO2+BzP and BP3+BzP On the endpoint of 48 h, a synergistic action was observed for nano-TiO2+BP3 and nano-TiO2+BzP, but an antagonistic effect occurred in the mixture BP3+BzP. These findings suggest that nano-TiO2 can increase the toxicity of the mixture when combined with other compounds.

Personal care products reconnaissance in EVROTAS river (Greece): Water-sediment partition and bioaccumulation in fish.

Twenty-six common ingredients of personal care products (PCPs) in water, sediment and fish from the Evrotas River (Greece) were investigated. Water sample analysis revealed the occurrence of twenty PCPs at concentrations ranging from 2.8 to 2031.0 ng l-1, the maximum corresponding to the endocrine disrupting UV filter benzophenone 3 (BP3). In sediment samples, six compounds were found to be adsorbed, the highest concentration being that of 4-methylbenzylidene camphor (4MBC, 1400.4 ng g-1 dw). Evrotas cyprinid fish (Squalius keadicus) showed a high accumulation potential for these chemicals, 100% detection frequency with maximum concentration that of benzophenone 2 (BP2, 41.9 ng g-1 dw). These data allowed estimating the distribution coefficients sediment-water (DCs-w) and the bioaccumulation factors (BAFs) of the pollutants investigated. Calculated rates revealed that benzophenone 1 (BP1), 4-hydroxybenzophenone (4HB) and ethyl-4-(dimethyl-amino)benzoate (EtPABA) have a strong tendency to adsorb onto the sediments, showing high DCs-w, i.e. 8.2E + 4 l g-1, 6.7E + 4 l g-1 and 5.7E + 3 l g-1, respectively. BFAs were only estimated for 5-methyl benzotriazole (MeBT), the compound having paired data from fish and water. The obtained values (range 2.0E + 2 l g-1-3.8E + 3 l g-1), indicated MeBT's strong bioaccumulation. Risk assessment of the investigated compounds for several aquatic organisms indicated a high ecological risk (HQ > 1) for BP3 and medium ecological risk (HQ ~ 0.5) for ODPABA.

On-line solid phase extraction-liquid chromatography-tandem mass spectrometry for insect repellent residue analysis in surface waters using atmospheric pressure photoionization.

Insect repellents (IRs) are a group of organic chemicals whose function is to prevent the ability of insects of landing in a surface. These compounds have been found in the environment and may pose a risk to non-target organisms. In this study, an on-line solid phase extraction - high performance liquid chromatography-tandem mass spectrometry multiresidue method was developed using an atmospheric photoionization source (SPE-HPLC-(APPI)-MS/MS). The use of the APPI as an alternative ionization technique to electrospray (ESI) and atmospheric pressure chemical ionization (APCI) allowed expanding the range of analytical techniques suitable for the analysis of IRs, so far relied in gas chromatography. High sensitivity and precision was reached with method limits of quantification between 0.2 and 4.6 ng l-1 and interday and intraday precision equal or below 15%. The validated method was applied to the study of surface water samples from three European river basins with different flow regime (Adige River in Italy, Sava River in the Balkans, and Evrotas River in Greece). The results showed that two IRs (DEET and Bayrepel) were ubiquitous in the Sava and Evrotas basins, reaching concentrations as high as 105 µg l-1 of Bayrepel in the Sava River, and 5 µg l-1 of DEET in the Evrotas River. Densely populated areas and effluent waste waters are pointed out as the responsible for this pollution. In the alpine river Adige, only three samples showed low levels of IRs (6.01-37.8 ng l-1). The concentrations measured were used to perform an environmental risk assessment based on the hazard quotients (HQs) estimation approach by using the chronic and acute eco-toxicity data available. The results revealed that despite the high frequency and eventually high concentrations of these IRs determined in the three basins, only few sites were at risk, with 1 < HQs < 3.3.

Determination of UV filters in human breast milk using turbulent flow chromatography and babies' daily intake estimation.

UV filters (UV-Fs) are a group of hormonally active chemical compounds used to protect against the deleterious effects of UVA and UVB solar radiation, which are currently present in most consumer goods (personal care products, plastics, fabrics, paints, etc). Last years the concern about these emerging contaminants has been on the rise, and increasing efforts are being taken in order to properly asses the hazard that the exposure to these compounds in the early stages of life may pose. In this study, a new method for the analysis of 11 UV-Fs residues in human breast milk samples has been developed. The method is based on turbulent flow chromatography coupled to liquid chromatography-tandem mass spectrometry (TFC-HPLC-MS/MS). The validated method was successfully applied to 79 human breast milk samples from mothers in Barcelona (Spain). Twenty-four per cent of the samples contained UV-Fs, with major contributors being oxybenzone (benzophenone 3, BP3), its metabolite 4,4'-dihydroxybenzophenone (4DHB), and UV320 showing maximum concentrations of 779.9, 73.3, and 523.6ngg-1 milk, respectively. Additionally, the plastic containers of the milks were also analysed, revealing high concentrations of BP3 and 4DHB, up to 10.6µgg-1 plastic. The calculated mean ΣUV-Fs were useful to estimate the daily intake (EDI) by babies, which were 69.1µg d-1kg-1 body weight.

Occurrence of organic UV filters and metabolites in lebranche mullet (Mugil liza) from Brazil.

UV filters (UV-Fs) constitute a heterogeneous group of chemicals used as protection against the effects of UV radiation, widely used in all sort of goods and ubiquitous in the environment. The presence of these chemicals in fish is a matter of concern, because many UV-Fs display hormonal activity. In this study, muscle, gills, and liver from 11 Mugil liza individuals from the highly urbanized Guanabara Bay (Rio de Janeiro, Brazil) were analysed in order to detect eight UV-Fs and metabolites (4-dihydroxybenzophenone [BP1] (2-hydroxy-4-methoxybenzophenone [BP3], 4-methylbenzylidiene camphor [4MBC], ethylhexyl methoxycinnamate [EHMC], ethylhexyl dimethyl p-aminobenzoic acid [ODPABA], octocrylene [OC], 4-hydroxybenzophenone [4HB], and 4,4'-dihydroxybenzophenone [4DHB]) using liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Results showed that both target UV-Fs and metabolites were ubiquitous in the analysed tissues. Lower concentrations were observed in muscle and gills (3.07-31.6ngg-1 dry weight (dw)), whereas in liver significant amounts of metabolites (5.47-451ngg-1 dw) were present. With the concentrations determined in the fish, an estimation of the daily intake revealed that consumption of muscle in the diet represent from 0.3 to 15.2ng UV-Fs (kg body weight-1) d-1, higher than those reported in fish for selected persistent organic pollutants (POPs).

A Potential New Threat to Wild Life: Presence of UV Filters in Bird Eggs from a Preserved Area.

The present study uses bird eggs of seven wild species as a biomonitoring tool for sunscreens occurrence. Seven UV filters (UV-Fs), including 3 hydroxy-metabolites of oxybenzone (benzophenone 3, BP3) were characterized in unhatched eggs from Doñana Natural Space (Spain). High frequency of detection was observed for all UV-Fs, ranging from 95% to 100%. The oxybenzone metabolite 4-hydroxybenzophenone (4HB) was ubiquitous at concentrations in the range 12.0-3348 ng g-1 dry weight (dw). The parent compound, oxybenzone, was also present in all samples at lower concentrations (16.9-49.3 ng g-1 dw). Due to the three BP3's metabolites, benzophenone 1 (BP1), 4HB, and 4,4'-dihydroxybenzophenone (4DHB) presence in unhatched eggs, it can be inferred that parent compounds are absorbed into the bird through the upper gut and the OH-derivatives formed are transferred by the mother to the egg before the lying. White stork (Ciconia ciconia) and western marsh harrier (Circus aeruginosus) were the most contaminated species, with mean total UV-Fs concentrations of 834 and 985 ng g-1 dw, respectively. Results evidenced that biomagnification process across the bird species studied cannot be ruled out.

Sediments as a sink for UV filters and benzotriazoles: the case study of Upper Iguaçu watershed, Curitiba (Brazil).

Ingredients in home and personal care products, including UV filters and benzotriazoles, are high production volume chemicals extensively used in our daily life, despite several studies revealed their potential eco-toxicity and endocrine-disrupting capacity. Due to some features, such as high lipophilicity, low degradability, and persistence of many of these compounds, sediments can be considered a sink for them in the aquatic environment. In the present study, nine organic UV filters and three benzotriazoles were investigated for the first time in sediments from four urban rivers in Brazil. The contaminants were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results revealed that octocrylene (OC), etylhexyl-methoxycinnamate (EHMC), benzophenone-3 (oxybenzone, BP3), and benzotriazole (BZT) were the predominant compounds adsorbed on the sediments, with concentrations ranging from 5.6 to 322.2 ng g-1 dry weight. The results reported in this work constitute the first data on the accumulation of polar benzotriazoles and lipophilic organic UV filters in sediments from Brazil.

UV filters and benzotriazoles in urban aquatic ecosystems: The footprint of daily use products.

The increased use of beauty and other daily use products, in particular those containing UV filters (UV-Fs) and benzotriazoles, results in their introduction in significant amounts into the aquatic environment. In this study, we aim to assess the occurrence and impact of UV-Fs and benzotriazoles in aquatic ecosystems in the metropolitan area of Barcelona, Spain. River water samples from the Llobregat and Besòs Rivers were analysed together with sediment, suspended particulate matter, and wastewater samples from 6 wastewater treatment plants (WWTPs) along their basins. The analysis of 6 UV-Fs and 2 benzotriazoles in water samples was performed using an automatized on-line solid phase extraction coupled to liquid chromatography tandem mass spectrometry (SPE-HPLC-MS/MS) method. The analysis of the target compounds in the suspended solids and in the sediments was performed by HPLC-MS/MS. The analysis of the water samples showed the ubiquitous presence of UV-Fs. Benzotriazole (BZT; partition coefficient octanol-water Log Kow=1.23) and methylbenzotriazole (MeBZT; Log Kow=1.89) had the highest levels in both river water and wastewater. Removal rates in the selected WWTPs were highly variable (4-100%). Concentrations of lipophilic UV-Fs (Log Kow 4.95-7.53) in suspended particulate matter from wastewaters were high (up to 1,031,868.2ngg-1dry weight (dw)), whereas in sediment the concentrations were always below 300ng g-1 dw. The risk assessment expressed in terms of hazard quotients (HQs) revealed that most UV-Fs were not likely to produce adverse ecotoxicological effects against the living organisms assayed in river waters and influent wastewaters at the concentrations observed. However, HQs above 1 were obtained for BZT and MeBZT in effluent wastewaters discharged to the river.

Contamination sources and distribution patterns of pharmaceuticals and personal care products in Alpine rivers strongly affected by tourism.

Knowledge regarding the impact of tourism on the emergence of pharmaceuticals and personal care products (PPCPs) in Alpine river waters is limited and scarce. Therefore, a study on the occurrence patterns and spatiotemporal variability of 105 PPCPs in an Alpine river basin located in the Trentino-Alto Adige region (North-Eastern Italy) has been conducted. We observed that the total concentration of analyzed PPCPs was generally higher in all sampling sites during winter than in the summer. The analysis of tourist data revealed that during both sampling campaigns the number of tourists was lower in the downstream sites in comparison with the upstream area of the basin (Val di Sole). Particularly, sampling sites located near important tourist resorts have shown the highest abundance of the PPCPs during winter, being analgesics/anti-inflammatories, antihypertensives and antibiotics the most abundant pharmaceutically active compounds (PhACs). Diclofenac showed the highest concentration amongst PhACs, reaching concentrations up to 675ngL-1 in the sampling site situated downstream of the Tonale wastewater treatment plant (WWTP). Antihypertensives were found at concentrations >300ngL-1, while antibiotics were quantified up to 196ngL-1, respectively. Amongst personal care products (PCPs), the most abundant compound was octyl-dimethyl-p-aminobenzoic acid (ODPABA) with concentrations reaching up to 748ngL-1 in the sampling site situated within the Rotaliana district. In general, concentrations and detection frequencies were higher in water than in the sediment samples. The most frequently detected PhACs in sediments from both sampling campaigns were antibiotics, while amongst PCPs in sediments, octocrylene (OC) showed the highest concentration in both sampling campaigns. As a result, this study highlights the potential impact of tourism on the water quality of the Alpine aquatic ecosystems.

Ecological risk assessment associated to the removal of endocrine-disrupting parabens and benzophenone-4 in wastewater treatment.

The occurrence of four widely used and endocrine disrupting parabens (PBs) (methylparaben, propylparaben, butylparaben and benzylparaben) and a polar UV filter (benzophenone-4) were determined in influent and effluent wastewater from the 19 major wastewater treatment plants (WWTPs) of Catalonia, Spain. For their analysis an on-line solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-HPLC-MS/MS) method was developed and validated. Laboratory analysis revealed high levels for both PBs and BP4, with maximum concentrations of 5700ngL(-1) and 1806ngL(-1), respectively, in influent samples, and 137ngL(-1) and 1080ngL(-1), respectively in effluent wastewaters. Removal rates (RE%) for the target compounds in each WWTPs were calculated. RE% for parabens were almost 100%, whereas for BP4 values where in the range 5-91%. The half-life time (t1/2), hydraulic retention time (HRT), and annual mass load (ML) for each facility was estimated. Results indicated that there was no clear influence of HRT on the RE% of BP4. MLs for BP4 were in the range 0.9-110.1kgy(-1), with the highest values in the most populated areas. Finally, a risk assessment, estimated in terms of hazard quotients (HQs), was carried out for aquatic biota. HQs for the target compounds in effluent wastewaters indicated a negligible effect, whereas for some influent wastewaters' HQs pointed out that some species are at risk.

Determination of parabens and benzophenone-type UV filters in human placenta. First description of the existence of benzyl paraben and benzophenone-4.

UV filters and parabens (PBs) are chemicals used in daily personal care and hygiene products to protect materials and humans from the adverse effects of UV radiation and to preserve the integrity of the formulation, respectively. Several studies highlight their widespread environmental occurrence and endocrine disrupting effects. However, little is known about human exposure to these compounds. The objective of this study was to investigate the exposure of human embryos and foetuses to endocrine disrupting UV filters and PBs. Placentas from volunteer mothers in Barcelona were collected at delivery after informed, written consent by the pregnant women. UV filters and parabens were analysed by liquid chromatography-tandem mass spectrometry. The excellent performance of the method allowed measuring the target compounds in human placental tissue at low ng/g fresh weight level. The detection frequency of the selected compounds was in the range 17-100%. Benzophenone-1, methyl paraben, butyl paraben and benzyl paraben were detected in all samples. The highest measured concentration corresponded to methyl paraben, 11.77ng/g fresh weight. Reported concentrations of benzophenone-4 and benzyl paraben constitute the first evidence about their accumulation in placenta. The results obtained corroborate that foetuses are exposed to a wide diversity of UV filters and PBs via the placenta.

Single and joint ecotoxicity data estimation of organic UV filters and nanomaterials toward selected aquatic organisms. Urban groundwater risk assessment.

The hazardous potential of organic UV filters (UV-Fs) is becoming an issue of great concern due to the widespread application of these compounds in most daily-use goods, such as hygiene and beauty products. Nanomaterials (NMs) have also been used in personal care products (PCPs) for many years. Nowadays, both classes of chemicals are considered environmental emerging contaminants. Despite some studies performed in vitro and in vivo reported adverse effects of many UV-Fs on the normal development of organisms, there is scarce data regarding acute and chronic toxicity. The aim of the present study was to determine the EC50 values of selected UV-Fs using standardised toxicity assays on three aquatic species i.e. Daphnia magna, Raphidocelis subcapitata and Vibrio fischeri. EC50 values obtained were in the mgl(-1) range for all the species. The estimated toxicity data allowed us to assess the environmental risk posed by selected UV-Fs in urban groundwater from Barcelona (Spain). The calculated ecological risk indicated a negligible impact on the aquifer. Giving the increasing importance of studying mixtures of pollutants and due to the widespread presence of nanomaterials (NMs) in the aquatic environment, other objective of this work was to explore the response on D. magna after exposure to both binary combinations of UV-Fs among them and UV-F with NMs. In all cases but the nano-silver mixtures, joint toxicity was mitigated or even eradicated.

Removal of polar UV stabilizers in biological wastewater treatments and ecotoxicological implications.

The present study describes the development, validation and application of a fully automated analytical method based on on-line solid-phase extraction-liquid chromatography-tandem mass spectrometry (on line SPE-HPLC-MS/MS) to assess the removal efficiency in water works and the ecotoxicological implications derived of the two most used benzotriazole-class UV stabilizers (BZTs), namely 1H-benzotriazole (BZT) and 5-methyl-1H-benzotriazole (MeBZT). Influent and effluent wastewater samples from 20 wastewater treatment plants (WWTPs) were analyzed. Removal rates (RE%) and half-lives (t1/2) for each BZTs were calculated and correlated to the hydraulic retention time (HRT) of each plant. Both BZTs were detected in all influent and effluent samples (concentrations in the range 26.7 ng L(-1)-42.9 µg L(-1)), with the highest concentrations corresponding to MeBZT. Results indicated that both compounds were recalcitrant (RE% in the range 11.8-94.7%) and that no clear influence of HRT on removals could be drawn. Finally, the potential environmental risk posed by the levels of BZTs detected was evaluated calculating the hazard quotients (HQs) MeBZT was the only BZTs posing a risk to Vibrio fischeri, Daphnia galeata and Pimephales promelas.

Re-inoculation strategies enhance the degradation of emerging pollutants in fungal bioaugmentation of sewage sludge.

The use of Trametes versicolor has been partially successful in the removal of some pharmaceuticals from sewage sludge in laboratory-scale biopile systems. The application of two strategies for the re-inoculation of biomass was assessed during the fungal bioaugmentation of non-sterile sludge (42-d treatment) as an approach to improve the elimination of pharmaceuticals and other groups of emerging pollutants. Globally, the re-inoculation of biopiles with blended mycelium exerted a major effect on the removal of pharmaceuticals (86%), brominated-flame-retardants (81%) and UV filters (80%) with respect to the re-inoculation with additional lignocellulosic substrate colonized by the fungus (69-67-22%). The performance was better than that of the analogous non-re-inoculated systems that were assayed previously for the removal of pharmaceuticals. The results demonstrate the ability of T. versicolor to remove a wide spectrum of emerging micropollutants under non-sterile conditions, while re-inoculation appears to be a useful step to improve the fungal treatment of sludge.