Degradation and mineralization of anti-cancer drugs Capecitabine, Bicalutamide and Irinotecan by UV-irradiation and ozone.

The degradation of three anti-cancer drugs (ADs), Capecitabine (CAP), Bicalutamide (BIC) and Irinotecan (IRI), in ultrapure water by ozonation and UV-irradiation was tested in a bench-scale reactor and AD concentrations were measured through ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). A low-pressure mercury UV (LP-UV) lamp was used and degradation by UV (λ = 254 nm) followed pseudo-first order kinetics. Incident radiation in the reactor was measured via chemical actinometry using uridine. The quantum yields (φ) for the degradation of CAP, BIC and IRI were 0.012, 0.0020 and 0.0045 mol Einstein-1, respectively. Ozone experiments with CAP and IRI were conducted by adding ozone stock solution to the reactor either with or without addition of tert-butanol (t-BuOH) as radical quencher. Using this experimental arrangement, no degradation of BIC was observed, so a semi-batch setup was employed for the ozone degradation experiments of BIC. Without t-BuOH, apparent second order reaction rate constants for the reaction of the ADs with molecular ozone were determined to be 3.5 ± 0.8 ∙ 103 L mol-1 s-1 (CAP), 7.9 ± 2.1 ∙ 10-1 L mol-1 s-1 (BIC) and 1.0 ± 0.3 ∙ 103 L mol-1 s-1 (IRI). When OH-radicals (∙OH) were quenched, rate constants were virtually the same for CAP and IRI. For BIC, a significantly lower constant of 1.0 ± 0.5 ∙ 10-1 L mol-1 s-1 was determined. Of the tested substances, BIC was the most recalcitrant, with the slowest degradation during both ozonation and UV-irradiation. The extent of mineralization was also determined for both processes. UV irradiation was able to fully degrade up to 80% of DOC, ozonation up to 30%. Toxicity tests with Daphnia magna (D. magna) did not find toxicity for fully degraded solutions of the three ADs at environmentally relevant concentrations.

CALEOSIN 1 interaction with AUTOPHAGY-RELATED PROTEIN 8 facilitates lipid droplet microautophagy in seedlings.

Lipid droplets (LDs) of seed tissues are storage organelles for triacylglycerols (TAGs) that provide the energy and carbon for seedling establishment. In the major route of LD degradation (lipolysis), TAGs are mobilized by lipases. However, LDs may also be degraded via lipophagy, a type of selective autophagy, which mediates LD delivery to vacuoles or lysosomes. The exact mechanisms of LD degradation and the mobilization of their content in plants remain unresolved. Here, we provide evidence that LDs are degraded via a process morphologically resembling microlipophagy in Arabidopsis (Arabidopsis thaliana) seedlings. We observed the entry and presence of LDs in the central vacuole as well as their breakdown. Moreover, we show co-localization of AUTOPHAGY-RELATED PROTEIN 8b (ATG8b) and LDs during seed germination and localization of lipidated ATG8 (ATG8-PE) to the LD fraction. We further demonstrate that structural LD proteins from the caleosin family, CALEOSIN 1 (CLO1), CALEOSIN 2 (CLO2), and CALEOSIN 3 (CLO3), interact with ATG8 proteins and possess putative ATG8-interacting motifs (AIMs). Deletion of the AIM localized directly before the proline knot disrupts the interaction of CLO1 with ATG8b, suggesting a possible role of this region in the interaction between these proteins. Collectively, we provide insights into LD degradation by microlipophagy in germinating seeds with a particular focus on the role of structural LD proteins in this process.

Green composites based on volcanic red algae Cyanidiales, cellulose, and coffee waste biomass modified with magnetic nanoparticles for the removal of methylene blue.

In this paper, green nanocomposites based on biomass and superparamagnetic nanoparticles were synthesized and used as adsorbents to remove methylene blue (MB) from water with magnetic separation. The adsorbents were synthesized through the wet co-precipitation technique, in which iron-oxide nanoparticles coated the cores based on coffee, cellulose, and red volcanic algae waste. The procedure resulted in materials that could be easily separated from aqueous solutions with magnets. The morphology and chemical composition of the nanocomposites were characterized by SEM, FT-IR, and XPS methods. The adsorption studies of MB removal with UV-vis spectrometry showed that the adsorption performance of the prepared materials strongly depended on their morphology and the type of the organic adsorbent. The adsorption studies presented the highest effectiveness in neutral pH with only a slight effect on ionic strength. The MB removal undergoes pseudo-second kinetics for all adsorbents. The maximal adsorption capacity for the coffee@Fe3O4-2, cellulose@Fe3O4-1, and algae@Fe3O4-1 is 38.23 mg g-1, 41.61 mg g-1, and 48.41 mg g-1, respectively. The mechanism of MB adsorption follows the Langmuir model using coffee@Fe3O4 and cellulose@Fe3O4, while for algae@Fe3O4 the process fits to the Redlich-Peterson model. The removal efficiency analysis based on UV-vis adsorption spectra revealed that the adsorption effectiveness of the nanocomposites increased as follows: coffee@Fe3O4-2 > cellulose@Fe3O4-1 > algae@Fe3O4-1, demonstrating an MB removal efficiency of up to 90%.

Mesocosm experiment to determine the contribution of adsorption, biodegradation, hydrolysis and photodegradation in the attenuation of antibiotics at the water sediment interface.

To understand the fate of antibiotics in the aquatic environment, we need to evaluate to which extent the following processes contribute to the overall antibiotic attenuation: adsorption to river sediment, biodegradation, hydrolysis and photodegradation. A laboratory scale mesocosm experiment was conducted in 10 L reactors filled with river sediment and water. The reactors were spiked with four classes of antibiotics (fluoroquinolones, macrolides, sulfonamides, tetracyclines), as well as clindamycin and trimethoprim. The experimental-set-up was designed to study the attenuation processes in parallel in one mesocosm experiment, hence also considering synergetic effects. Our results showed that antibiotics belonging to the same class exhibited similar behavior. Adsorption was the main attenuation process for the fluoroquinolones and tetracyclines (44.4 to 80.0 %). For the sulfonamides, biodegradation was the most frequent process (50.2 to 65.1 %). Hydrolysis appeared to be significant only for tetracyclines (12.6 to 41.8 %). Photodegradation through visible light played a minor role for most of the antibiotics - fluoroquinolones, sulfonamides, and trimethoprim (0.7 to 24.7 %). The macrolides were the only class of antibiotics not affected by the studied processes and they persisted in the water phase. Based on our results, we propose to class the antibiotics in three groups according to their persistence in the water phase. Fluoroquinolones and tetracyclines were non-persistent (half-lives shorter than 11 d). Chlorotetracycline, sulfapyridine and trimethoprim showed a moderate persistence (half-lives between 12 and 35 d). Due to half-lives longer than 36 d sulfonamides and clindamycin were classified as persistent.

Removal of pharmaceuticals from nitrified urine by adsorption on granular activated carbon.

Nitrification and distillation of urine allow for the recovery of all nutrients in a highly concentrated fertilizer solution. However, pharmaceuticals excreted with urine are only partially removed during these two process steps. For a sustainable and safe application, more extensive removal of pharmaceuticals is necessary. To enhance the pharmaceutical removal, which is already occurring during urine storage, nitrification and distillation, an adsorption column with granular activated carbon (GAC) can be included in the treatment train. We executed a pilot-scale study to investigate the adsorption of eleven indicator pharmaceuticals on GAC. During 74 days, we treated roughly 1000 L of pre-filtered and nitrified urine spiked with pharmaceuticals in two flow-through GAC columns filled with different grain sizes. We compared the performance of these columns by calculating the number of treated bed volumes until breakthrough and carbon usage rates. The eleven spiked pharmaceuticals were candesartan, carbamazepine, clarithromycin, diclofenac, emtricitabine, hydrochlorothiazide, irbesartan, metoprolol, N4-acetylsulfamethoxazole, sulfamethoxazole and trimethoprim. At the shortest empty bed contact time (EBCT) of 25 min, immediate breakthrough was observed in both columns shortly after the start of the experiments. Strong competition by natural organic material (NOM) could have caused the low pharmaceutical removal at the EBCT of 25 min. At EBCTs of 70, 92 and 115 min, more than 660 bed volumes could be treated until breakthrough in the column with fine GAC. The earliest breakthrough was observed for candesartan and clarithromycin. On coarse GAC, only half the number of bed volumes could be treated until breakthrough compared to fine GAC. The probable reason for the later breakthrough with fine GAC is the smaller intraparticle diffusive path length. DOC and UV absorbance measurements at 265 nm indicated that both parameters can be used as indicators for the breakthrough of pharmaceuticals. In contrast to pharmaceuticals and DOC, the nutrient compounds ammonium, nitrate, phosphate, potassium and sulfate were not removed significantly. A comparison with literature values suggests that the amount of GAC needed to remove pharmaceuticals from human excreta could be reduced by nearly two orders of magnitude, if urine were treated on site instead of being discharged and treated in a centralized wastewater treatment plant.

Decay of elevated antibiotic resistance genes in natural river sediments after sedimentation of wastewater particles.

Large-scale wastewater treatment plants (WWTPs) discharge hundreds of tons of total suspended solids (TSS) into surface waters every year. Additionally, a comparable amount is released by sewer overflows during heavy rain events in case of combined sewer systems. Along with sedimentation, particle-attached microorganisms and their antibiotic resistance genes (ARGs) are herewith transported to the riverbed of the receiving water. To better understand the dynamics of this process, a particulate wastewater fraction was added into batch reactors, which were previously filled with natural river sediments and tap water. In parallel, antibiotics (ABs) (erythromycin, tetracycline, ciprofloxacin, roxithromycin, penicillin V, and sulfamethoxazole) were spiked to investigate their capability to select for resistance. The abundance of six ARGs (ermB, tetM, blaTEM, sul1, CTX-M-32, and qnrS) as well as total bacteria (16S rDNA) was monitored in waters and in sediments for a duration of two months using quantitative PCR. Despite a continuous exposure to ABs (5 µg/L each), the abundance of ARGs remained unaffected. Addition of wastewater particles resulted in a sudden and strong increase of ARGs in waters (3-5 log units) and sediments (1-4 log units), however, elevated ARGs underwent a particular and complete decay. Our results indicate that the increased ARG abundances in receiving rivers are the result of a continuous import of ARGs from WWTP discharges or sewer overflow events. They further imply that elevated ARGs do not persist in receiving rivers, if this continuous import is removed. This seems to be the case merely for ARGs introduced by wastewater, given that a stable background concentration of ARGs was observed for the native population.

Solar-mediated degradation of linezolid and tedizolid under simulated environmental conditions: Kinetics, transformation and toxicity.

Linezolid (LIN) and Tedizolid (TED) are representatives of oxazolidinone antibiotics of last resort with a strong efficacy against gram-positive bacteria. This study focused on their solar-mediated degradation to understand better their fate in aquatic environment, for the realistic concentrations in the range of 1 µg/L. Results showed that both antibiotics (ABs) are degradable by simulated sunlight (1 kW/m2), with half-lives of 32 and 93 h in ultrapure water, for LIN and TED, respectively. LIN showed similar photolytic behaviour in pure solution and in surface water, whereas sunlight enhanced the degradation of LIN in pure solutions, but not in surface water. Structure elucidation by liquid chromatography coupled to high resolution mass spectrometry provided information about seven transformation products for LIN and five for TED. The morpholinyl-ring was identified as the target site for most transformation reactions of LIN. TED was prone to oxidation and cleavage of the oxazolidinone ring. Results of a growth inhibition test on Bacillus subtilis exposed to UV light showed antibacterial efficacy of transformation products of LIN and no significant efficacy of degradation products of TED for the concentration range of 100 µg/L-10 mg/L of parent compounds. Photolytically treated solutions of the ABs maintained their inhibitory effect on the bioluminescence of Aliivibrio fischeri.

Impact of the particulate matter from wastewater discharge on the abundance of antibiotic resistance genes and facultative pathogenic bacteria in downstream river sediments.

Wastewater treatment plants (WWTPs) are point sources for both, the release of antibiotic resistance genes (ARGs) and the discharge of antibiotics (ABs) into the environment. While it is well established that ARGs emission by WWTPs leads to an ARGs increase in receiving rivers, also the role of sub-inhibitory AB concentrations in this context is being discussed. However, the results obtained in this study suggest that, at environmental concentrations, ABs do not have an effect on resistance selection. Instead, we emphasize the significance of ARG transport and, in that respect, highlight the relevance of wastewater particles and associated microorganisms. We can show that ARGs (ermB, blaTEM,tetM, qnrS) as well as facultative pathogenic bacteria (FPB) (enterococci, Pseudomonas aeruginosa, Acinetobacter baumannii) inside the particulate fraction of WWTP effluent are very likely to remain in the riverbed of the receiving water due to sedimentation. Moreover, ARG and FPB abundances measured in the particulate fraction strongly correlated with the delta ARG and FPB abundances measured in the receiving river sediment (downstream compared to upstream) (R2 = 0.93, p < 0.05). Apparently, the sheer amount of settleable ARGs and FPB from WWTP effluent is sufficient, to increase abundances in the receiving riverbed by 0.5 to 2 log units.

Photolysis of four ß­lactam antibiotics under simulated environmental conditions: Degradation, transformation products and antibacterial activity.

߭Lactam antibiotics are among the most widely used antibiotics in human medicine and their effects on the aquatic environment - concerning bacterial resistance - are controversially discussed. This study focused on the photolysis of the four ߭lactam antibiotics - amoxicillin, ampicillin, penicillin V and piperacillin - under simulated environmental conditions. It was observed that all investigated ߭lactam antibiotics are photolytically degradable by simulated sunlight (1 kW/m2) with half-lives between 3.2 and 7.0 h. Structure elucidation of transformation products performed with liquid chromatography coupled to high resolution mass spectrometry showed that the hydrolysis of the ߭lactam ring is the primary transformation reaction, followed by the elimination of carboxylic and dimethyl thiazolidine carboxylic acid. Growth inhibition tests on Bacillus subtilis showed the loss of bactericide activity of irradiated solutions of amoxicillin, ampicillin and piperacillin, suggesting the transformation of the ߭lactam ring is responsible for the antibiotic effect. In contrast, the solutions of penicillin V did not show any decline of the antibacterial activity after photolytic degradation, probably due to the formation of still active epimers.

Evaluation of a full-scale wastewater treatment plant upgraded with ozonation and biological post-treatments: Abatement of micropollutants, formation of transformation products and oxidation by-products.

To protect the ecosystem and drinking water resources in Switzerland and in the countries of the downstream catchments, a new Swiss water protection act entered into force in 2016 aiming to reduce the discharge of micropollutants from wastewater treatment plants (WWTPs). As a consequence, selected WWTPs must be upgraded by an advanced treatment for micropollutant abatement with suitable and economic options such as (powdered) activated carbon treatment or ozonation. WWTP Neugut (105'000 people equivalent) was the first WWTP in Switzerland to implement a long-term full-scale ozonation. Differing specific ozone doses in the range of 0.35-0.97 g O3/g DOC were applied to determine the adequate ozone dose to fulfill the requirements of the Swiss water protection act. Based on this assessment, a specific ozone dose of 0.55 g O3/g DOC is recommended at this plant to ensure an average abatement of the twelve selected indicator substances by ≥80% over the whole treatment. A monitoring of 550 substances confirmed that this dose was very efficient to abate a broad range of micropollutants by >79% on average. After ozonation, an additional biological post-treatment is required to eliminate possible negative ecotoxicological effects generated during ozonation caused by biodegradable ozonation transformation products (OTPs) and oxidation by-products (OBPs). Three biological treatments (sand filtration, moving bed, fixed bed) and granular activated carbon (GAC, fresh and pre-loaded) filtration were evaluated as post-treatments after ozonation. In parallel, a fresh GAC filter directly connected to the effluent of the secondary clarifier was assessed. Among the three purely biological post-treatments, the sand filtration performed best in terms of removal of dissolved organic carbon (DOC), assimilable organic carbon (AOC) and total suspended solids (TSS). The fresh activated carbon filtration ensured a significant additional micropollutants abatement after ozonation due to sorption. The relative abatement of the indicator substances ranged between 20 and 89% after 27'000 bed volumes (BV) and was still substantial at 50'000 BV. In an identical GAC filter running in parallel and being fed with the effluent of the secondary clarifier, the elimination was less efficient. Seven primary OTPs (chlorothiazide and six N-oxides) formed during ozonation could be quantified thanks to available reference standards. Their concentration decreased with increasing specific ozone doses with the concomitant formation of other OTPs. The seven OTPs were found to be stable compounds and were not abated in the biological post-treatments. They were sorbed in the fresh GAC filter, but less efficiently than the corresponding parent compounds. Two OBPs, bromate (BrO3-) and N-nitrosodimethylamine (NDMA), were formed during ozonation but did not exceeded 5 µg/L for bromate and 30 ng/L for NDMA at the recommended specific ozone dose of 0.55 g O3/g DOC. NDMA was well abated in all post-treatments (minimum 41% during fixed bed filtration, maximum 83% during fresh GAC filtration), while bromate was very stable as expected.

Effect of operational and water quality parameters on conventional ozonation and the advanced oxidation process O3/H2O2: Kinetics of micropollutant abatement, transformation product and bromate formation in a surface water.

The efficiency of ozone-based processes under various conditions was studied for the treatment of a surface water (Lake Zürich water, Switzerland) spiked with 19 micropollutants (pharmaceuticals, pesticides, industrial chemical, X-ray contrast medium, sweetener) each at 1 µg L-1. Two pilot-scale ozonation reactors (4-5 m3 h-1), a 4-chamber reactor and a tubular reactor, were investigated by either conventional ozonation and/or the advanced oxidation process (AOP) O3/H2O2. The effects of selected operational parameters, such as ozone dose (0.5-3 mg L-1) and H2O2 dose (O3:H2O2 = 1:3-3:1 (mass ratio)), and selected water quality parameters, such as pH (6.5-8.5) and initial bromide concentration (15-200 µg L-1), on micropollutant abatement and bromate formation were investigated. Under the studied conditions, compounds with high second-order rate constants kO3>104 M-1 s-1 for their reaction with ozone were well abated (>90%) even for the lowest ozone dose of 0.5 mg L-1. Conversely, the abatement efficiency of sucralose, which only reacts with hydroxyl radicals (OH), varied between 19 and 90%. Generally, the abatement efficiency increased with higher ozone doses and higher pH and lower bromide concentrations. H2O2 addition accelerated the ozone conversion to OH, which enables a faster abatement of ozone-resistant micropollutants. Interestingly, the abatement of micropollutants decreased with higher bromide concentrations during conventional ozonation due to competitive ozone-consuming reactions, except for lamotrigine, due to the suspected reaction of HOBr/OBr- with the primary amine moieties. In addition to the abatement of micropollutants, the evolution of the two main transformation products (TPs) of hydrochlorothiazide (HCTZ) and tramadol (TRA), chlorothiazide (CTZ) and tramadol N-oxide (TRA-NOX), respectively, was assessed by chemical analysis and kinetic modeling. Both selected TPs were quickly formed initially to reach a maximum concentration followed by a decrease of their concentrations for longer contact times. For the studied conditions, the TP's concentrations at the outlet of the reactors ranged from 0 to 61% of the initial parent compound concentration, CTZ being a more persistent TP against further oxidation than TRA-NOX. Finally, it was demonstrated in both reactors that the formation of bromate (BrO3-), a potentially carcinogenic oxidation by-product, could be controlled by H2O2 addition with a general improvement on micropollutant abatement. Post-treatment by granular activated carbon (GAC) filtration enabled the reduction of micropollutants and TPs concentrations but no changes in bromate were observed. The combined algae assay showed that water quality was significantly improved after oxidation and GAC post-treatment, driven by the abatement of the spiked pesticides (diuron and atrazine).

Oxidation of cetirizine, fexofenadine and hydrochlorothiazide during ozonation: Kinetics and formation of transformation products.

The efficiency of wastewater ozonation for the abatement of three nitrogen-containing pharmaceuticals, two antihistamine drugs, cetirizine (CTR) and fexofenadine (FXF), and the diuretic drug, hydrochlorothiazide (HCTZ), was investigated. Species-specific second-order rate constants for the reactions of the molecular, protonated (CTR, FXF) or deprotonated (HCTZ) forms of these compounds with ozone were determined. All three compounds are very reactive with ozone (apparent second order rate constants at pH 7: kO3,pH7 = 1.7·10(5) M(-1)s(-1), 8.5·10(4) M(-1)s(-1) and 9.0·10(3) M(-1)s(-1) for CTR, HCTZ and FXF, respectively). Transformation product (TP) structures were elucidated using liquid chromatography coupled with high-resolution tandem mass spectrometry, including isotope-labeled standards. For cetirizine and hydrochlorothiazide 8 TPs each and for fexofenadine 7 TPs were identified. The main TPs of cetirizine and fexofenadine are their respective N-oxides, whereas chlorothiazide forms to almost 100% from hydrochlorothiazide. In the bacteria bioluminescence assay the toxicity was slightly increased only during the ozonation of cetirizine at very high cetirizine concentrations. The main TPs detected in bench-scale experiments were also detected in full-scale ozonation of a municipal wastewater, for >90% elimination of the parent compounds.

Degradation of Iodinated Contrast Media in Aquatic Environment by Means of UV, UV/TiO2 Process, and by Activated Sludge.

Iodinated contrast media (ICM), which are used for radiological visualization of human tissue and cardiovascular system, are poorly biodegradable; hence, new methods of their removal are sought. In this study, the effectiveness of selected X-ray ICM removal by means of UV and UV/TiO2 pretreatment processes from synthetic hospital wastewater was demonstrated. The following compounds were investigated: iodipamide, iohexol, and diatrizoate. The experiments were as follows: (i) estimated susceptibility of the ICM to decay by UV radiation in different aquatic matrices, (ii) determined an optimal retention time of hospital wastewater in the UV reactor, (iii) determined optimum TiO2 concentration to improve the effectiveness of the UV pretreatment, and (iv) investigated removal of ICM by combination of the photochemical and biological treatment methods. The quantum yields of selected ICM decay in deionized water (pH = 7.0) were established as 0.006, 0.004, and 0.029 for iohexol, diatrizoate, and iodipamide, respectively. Furthermore, the experiments revealed that diatrizoate and iohexol removal in the UV/TiO2 process is more efficient than in UV process alone. For diatrizoate, the removal efficiency equaled to 40 and 30 %, respectively, and for iohexol, the efficiency was 38 and 27 %, respectively. No significant increase in iodipamide removal in UV and UV/TiO2 processes was observed (29 and 28 %, respectively). However, highest removal efficiency was demonstrated in synthetic hospital wastewater with the combined photochemical and biological treatment method. The removal of diatrizoate and iohexol increased to at least 90 %, and for iodipamide, to at least 50 %.

A chemiluminescence flow immunosensor based on a porous monolithic metacrylate and polyethylene composite disc modified with protein G.

A generic, fast, sensitive and new type of flow immunosensor has been developed. The basis is a monolithic porous poly(glycidyl methacrylate-co-trimethylolpropane trimethacrylate) polymer disc modified with protein G, placed in a fountain type flow cell compartment, in close proximity to a photomultiplier tube (PMT). Analyte and HRP labelled analyte derivative (tracer) compete for anti-analyte antibody binding sites. The mixture is then injected into the flow immunosensor system where the formed analyte- and tracer-antibody complexes are trapped by the monolithic protein G disc. The amount of bound tracer, inversely related to the concentration of analyte in the sample, is determined in a second step by injection of luminol, p-iodophenol and H2O2, generating enhanced chemiluminescence (CL) with horseradish peroxidase (HRP). A third and final step is need for regeneration of the protein G disc so that a new analysis cycle can take place. The performance of the disc immunosensor system was compared with a one step continuous flow injection immunoassay (FIIA) system, using the same reagents and a protein G column, in terms of assay sensitivity and influence of matrix effects from various water samples (millipore-, tap- and surface water). The detection limit for the analyte atrazine in PBS and surface water (SW) was 0.208 +/- 0.004 microg l(-1) (PBS) and 0.59 +/- 0.120 microg l(-1) (SW) for the FIIA and 0.033 +/- 0.003 microg l(-1) (PBS) and 0.038+/-0.003 microg l(-1) (SW) for the disc immunosensor. Statistical comparison of the two systems shows that the disc immunosensor results were significantly less influenced by the sample matrix, which is explained by the fact that the sample in the FIIA arrives simultaneously with the matrix to the detector, whereas these are separated in time in the disc immunosensor system.