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An anaerobic stable mixed culture dominated by bacteria belonging to the genera Dehalobacterium, Acetobacterium, Desulfovibrio, and Wolinella was used as a model to study the microbial interactions during DCM degradation. Physiological studies indicated that DCM was degraded in this mixed culture at least in a three-step process: i) fermentation of DCM to acetate and formate, ii) formate oxidation to CO 2 and H 2 , and iii) H 2 /CO 2 reductive acetogenesis. The 16S rRNA gene sequencing of cultures enriched with formate or H 2 showed that Desulfovibrio was the dominant population followed by Acetobacterium, but sequences representing Dehalobacterium were only present in cultures amended with DCM. Nuclear magnetic resonance analyses confirmed that acetate produced from 13 C-labelled DCM was marked at the methyl ([2- 13 C]acetate), carboxyl ([1- 13 C]acetate), and both ([1,2- 13 C]acetate) positions, which is in accordance to acetate formed by both direct DCM fermentation and H 2 /CO 2 acetogenesis. The inhibitory effect of ten different co-contaminants frequently detected in groundwaters on DCM degradation was also investigated. Complete inhibition of DCM degradation was observed when chloroform, perfluorooctanesulfonic acid, and diuron were added at 838, 400, and 107 μM, respectively. However, the inhibited cultures recovered the DCM degradation capability when transferred to fresh medium without co-contaminants. Findings derived from this work are of significant relevance to provide a better understanding of the synergistic interactions among bacteria to accomplish DCM degradation as well as to predict the effect of co-contaminants during anaerobic DCM bioremediation in groundwater., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The removal of 27 endocrine-disrupting compounds and related compounds (suspect effect) from a reverse osmosis concentrate using an alternative decontamination method based on a fungal treatment involving Trametes versicolor was assessed. In addition to chemical analysis, the toxicity of the treated water during the treatment was monitored using a bioluminescence inhibition test and estrogenic and anti-estrogenic tests. The compounds 1H-benzotriazole (BTZ) and two tolyltriazoles (TTZs), 4-methyl-1H-benzotriazole (4-MBTZ) and 5-methyl-1H-benzotriazole (5-MBTZ), were present in the reverse osmosis concentrate at the highest concentrations (7.4 and 12.8 μg L -1 , respectively) and were partially removed by the fungal treatment under sterile conditions (58% for BTZ and 92% for TTZs) and non-sterile conditions, although to lesser extents (32% for BTZ and 50% for TTZs). Individual biotransformation studies of BTZ and the TTZs by T. versicolor in a synthetic medium and further analysis via on-line turbulent flow chromatography coupled to an HRMS-Orbitrap allowed the tentative identification of the transformation products (TPs). Six TPs were postulated for BTZ, two TPs were postulated for 4-MBTZ, and four TPs were postulated for 5-MBTZ. Most of these TPs are suggested to have been generated by conjugation with some sugars and via the methylation of the triazole group. Only TP 148 A, postulated to be derived from the biotransformation of BTZ, was observed in the effluent of the bioreactor treating the reverse osmosis concentrate., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

This study investigated the removal of antibiotic ciprofloxacin during the treatment of real wastewater using high rate algal ponds (HRAP). When spiked at 2 mg/L into primary domestic wastewater, ciprofloxacin (CPX) was efficiently removed from laboratory scale photobioreactors continuously operated under various durations of artificial illumination and hydraulic residence times. Subsequent batch tests conducted with reactor microcosms showed CPX removal was mainly caused by photodegradation during daytime, and sorption to biomass during night time. These findings were confirmed during an experiment conducted in a 1000 L pilot HRAP operated outdoors, as well as during outdoor batch assays conducted using pilot HRAP microcosms. While these results highlight a potentially interesting treatment capacity in comparison to conventional biological treatment, further research must confirm these findings at relevant pollutant concentration (ng-μg/L) and determine the fate and potential toxicity of degradation products., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

This work presents a study about the elimination of anticancer drugs, a group of pollutants considered recalcitrant during conventional activated sludge wastewater treatment, using a biological treatment based on the fungus Trametes versicolor. A 10-L fluidized bed bioreactor inoculated with this fungus was set up in order to evaluate the removal of 10 selected anticancer drugs in real hospital wastewater. Almost all the tested anticancer drugs were completely removed from the wastewater at the end of the batch experiment (8 days) with the exception of Ifosfamide and Tamoxifen. These two recalcitrant compounds, together with Cyclophosphamide, were selected for further studies to test their degradability by T. versicolor under optimal growth conditions. Cyclophosphamide and Ifosfamide were inalterable during batch experiments both at high and low concentration, whereas Tamoxifen exhibited a decrease in its concentration along the treatment. Two positional isomers of a hydroxylated form of Tamoxifen were identified during this experiment using a high resolution mass spectrometry based on ultra-high performance chromatography coupled to an Orbitrap detector (LTQ-Velos Orbitrap). Finally the identified transformation products of Tamoxifen were monitored in the bioreactor run with real hospital wastewater., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The widespread of emerging contaminants in the environment and their potential impact on humans is a matter of concern. White-rot fungi are cosmopolitan organisms able to remove a wide range of pharmaceuticals and personal care products (PPCP) through cometabolism (i.e. laccases and peroxidases) or detoxification mechanisms (i.e. cytochrome P450 system). However, the use of PPCP as carbon source for these organisms is largely unexplored. Here, we used carbon stable isotope tracer experiments to assess the fate of anti-inflammatory diclofenac (DCF) and UV filter benzophenone-3 (BP3) during degradation by Trametes versicolor. The comparison between carbon isotopic composition of emitted carbon dioxide from 13C-labelled DCF ([acetophenyl ring-13C6]-DCF) and 13C-BP3 ([phenyl-13C6]-BP3) versus their 12C-homologue compounds showed mineralization of about 45% and 10% of the 13C contained in their respective molecules after 9 days of incubation. The carbon isotopic composition of the bulk biomass and the application of amino acid-stable isotope probing (SIP) allowed distinguishing between incorporation of 13C from BP3 into amino acids, which implies the use of this emerging contaminant as carbon source, and major intracellular accumulation of 13C from DCF without implying the transformation of its labelled phenyl ring into anabolic products. A mass balance of 13C in different compartments over time provided a comprehensive picture of the fate of DCF and BP3 across their different transformation processes. This is the first report assessing biodegradation of PPCP by SIP techniques and the use of emerging contaminants as carbon source for amino acid biosynthesis., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Ketoprofen is a nonsteroidal anti-inflammatory drug that has been detected in the environment in the range of ng L(-1)-microg L(-1) due to its low degradability in some wastewater treatment plants. In this study, the use of the white-rot fungus Trametes versicolor to effectively degrade ketoprofen in a defined liquid medium was assessed. The fungus eliminated ketoprofen to nondetectable levels in 24h when it was added at 10mgL(-1) whereas at low concentration of 40microgL(-1) it was almost completely removed (95%) after 5h. Low extracellular laccase activity was detected in the T. versicolor cultures but the addition of the laccase-mediator system did not lead to ketoprofen oxidation. The cytochrome P-450 inhibitor 1-aminobenzotriazole reduced ketoprofen oxidation. These data suggest that the first oxidation step is cytochrome P450 mediated. During time-course degradation experiments, three intermediates were structurally elucidated and quantified by HPLC-DAD-MS and NMR: 2-[3-(4-hydroxybenzoyl)phenyl]-propanoic acid, 2-[(3-hydroxy(phenyl)methyl)phenyl]-propanoic acid, and 2-(3-benzoyl-4-hydroxyphenyl)-propanoic acid. The latter was reported for the first time in biological systems. After 7 d of incubation, only small amounts of 2-[(3-hydroxy(phenyl)methyl)phenyl]-propanoic acid (0.08mg) remained in the liquid medium in comparison with the initial ketoprofen dose (1.0mg), suggesting possible mineralization of ketoprofen., (Copyright 2009 Elsevier Ltd. All rights reserved.)

The feasibility of degrading triphenylmethane dyes by Trametes versicolor and laccase has been investigated for the following dyes: Acid Fuchsin, Brilliant Green 1, Basic Fuchsin, Methyl Green or Acid Green 16. The toxicity level of triphenylmethane dyes is linked to their basic character, but significant detoxification is obtained when there is biodegradation. Identification of enzymatic degradation products by (1)H NMR made it possible to propose a general rule for the laccase attack on triphenylmethane compounds. The enzyme completely degrades the molecular part of the canonical resonance substructures of dyes, because no N-substituted, mono-N and di-N,N substituted p-amine aromatic residues seem to be wholly degraded. No enzymatic degradation is observed in the cases of either the non-substituted or trisubstituted-N,N,Np-amine aromatic residues. On the other hand, for all the dyes tested, no aromatic residues are detected after fungal treatment; this means that T. versicolor is more capable of performing further degradation than is laccase. The results of this study demonstrated that compounds with a triphenylmethane structure can be degraded by T. versicolor even if they are highly toxic. The enzyme laccase plays an important role in the attack on the structure and a general rule for predicting which products would be obtained after the enzymatic treatment is suggested.

A screening using four white-rot fungi (Trametes versicolor, Irpex lacteus, Ganoderma lucidum and Phanerochaete chrysosporium) was performed on the degradation of 10 mg L(-1) of ibuprofen (IBU, anti-inflammatory), clofibric acid (CLOFI, lipid regulator) and carbamazepine (CARBA, antiepileptic/analgetic) after 7 d of incubation. Whereas IBU was extensively degraded by all the fungi tested, T. versicolor was the only strain able to degrade either CLOFI (approximately 91%) and CARBA (approximately 58%), although the latter was also degraded by G. lucidum (approximately 47%). In vitro experiments using manganese peroxidase and laccase-mediator system showed that extracellular fungal enzyme systems did not appear to play a role in the first step of degradation. However, our in vivo studies using the cytochrome P450 inhibitors 1-aminobenzotriazole and piperonyl butoxide suggested that the cytochrome P450 system may be involved in the first step of CLOFI and CARBA oxidation by T. versicolor. During the very early stages of IBU degradation by T. versicolor, two hydroxylated metabolites were detected: 1-hydroxy ibuprofen and 2-hydroxy ibuprofen. These byproducts were subsequently degraded by the fungus to 1,2-dihydroxy ibuprofen, that was not reported in biological systems to date. Furthermore, these results are of particular interest because CLOFI and CARBA are highly persistent in the aquatic environment and they pass unchanged or poorly transformed in wastewater treatment plants.

The white-rot fungus Trametes versicolor degraded trichloroethylene (TCE), a highly oxidized chloroethene, and produced 2,2,2-trichloroethanol and carbon dioxide as the main products of degradation, based on the results obtained using [13C]-TCE as the substrate. For a range of concentrations of TCE between 2 and 20 mg l(-1), 53% of the theoretical maximum chloride expected from complete degradation of TCE was observed. Laccase was shown to be induced by TCE, but did not appear to play a role in TCE degradation. Cytochrome P-450 appears to be involved in TCE degradation, as evidenced by marked inhibition of degradation of TCE in the presence of 1-aminobenzotriazole, a known inhibitor of cytochrome P-450. Our results suggested that chloral (trichloroacetaldehyde) was an intermediate of the TCE degradation pathway. The results indicate that the TCE degradation pathway in T. versicolor appears to be similar to that previously reported in mammals and is mechanistically quite different from bacterial TCE degradation.