On the capacity of ozonation to remove antimicrobial compounds, resistant bacteria and toxicity from urban wastewater effluents.

The degradation of erythromycin (ERY) and ethylparaben (EtP) in urban wastewater effluents at low concentration level during ozonation was investigated under different experimental conditions. Both substrates were rapidly eliminated within 2min at low ozone dose of 0.3mgL-1 and the experimental data were well fitted in the pseudo-first-order kinetic model. The ratio of HO- and O3-exposure (Rct) at the inherent pH was found to be 1.9×10-8. The degradation of ERY and EtP was pronounced at pH 8 compared to acidic pH conditions, while the degradation rate of both substrates was found to be matrix-depended. It was also shown that both O3- and HO-mediated pathways are involved in the degradation of EtP, whereas the saturated-rich structure of ERY renders it O3-recalcitrant. Under the optimum O3 dose, the BrO3- concentration was found to be lower than 10µgL-1. Five and fifteen transformation products were elucidated during ERY and EtP oxidation, respectively. The root and shoot inhibition can be attributed to the oxidation products formed upon dissolved effluent organic matter transformation. Escherichia coli harbouring resistance to ERY survived ozonation better than EtP-resistant E. coli. However, neither ERY- nor EtP-resistant E. coli were detected after 15min of ozonation.

Erythromycin oxidation and ERY-resistant Escherichia coli inactivation in urban wastewater by sulfate radical-based oxidation process under UV-C irradiation.

This study evaluates the feasibility of UV-C-driven advanced oxidation process induced by sulfate radicals SO4(.)- in degrading erythromycin (ERY) in secondary treated wastewater. The results revealed that 10 mg L(-1) of sodium persulfate (SPS) can result in rapid and complete antibiotic degradation within 90 min of irradiation, while ERY decay exhibited a pseudo-first-order kinetics pattern under the different experimental conditions applied. ERY degradation rate was strongly affected by the chemical composition of the aqueous matrix and it decreased in the order of: ultrapure water (kapp = 0.55 min(-1)) > bottled water (kapp = 0.26 min(-1)) > humic acid solution (kapp = 0.05 min(-1)) > wastewater effluents (kapp = 0.03 min(-1)). Inherent pH conditions (i.e. pH 8) yielded an increased ERY degradation rate, compared to that observed at pH 3 and 5. The contribution of hydroxyl and sulfate radicals (HO. and SO4(.)-) on ERY degradation was found to be ca. 37% and 63%, respectively. Seven transformation products (TPs) were tentatively elucidated during ERY oxidation, with the 14-membered lactone ring of the ERY molecule being intact in all cases. The observed phytotoxicity against the tested plant species can potentially be attributed to the dissolved effluent organic matter (dEfOM) present in wastewater effluents and its associated-oxidation products and not to the TPs generated from the oxidation of ERY. This study evidences the potential use of the UV-C/SPS process in producing a final treated effluent with lower phytotoxicity (<10%) compared to the untreated wastewater. Finally, under the optimum experimental conditions, the UV-C/SPS process resulted in total inactivation of ERY-resistant Escherichia coli within 90 min.

Transformation products and reaction pathways of carbamazepine during photocatalytic and sonophotocatalytic treatment.

This study examines the degradation of the antiepileptic carbamazepine (CBZ) by sonolysis, TiO2-based heterogeneous photocatalysis under UV-A and simulated solar irradiation, and by the combined use of UV-A and ultrasound irradiation (i.e. sonophotocatalysis) in demineralized water, ground water and effluent wastewater. The processes were compared with respect to substrate conversion rate and the extent of DOC reduction as a measure of mineralization. CBZ was degraded following a pseudo-first order kinetics. Sonophotocatalysis provided the highest rate of CBZ transformation over the time-course of the experiment while the degree of DOC removal in pure water was similar for all the studied treatments (around 40%), and always lower than CBZ conversion. This indicated that a considerable organic load remained in the treated solutions that could also be attributed to the presence of persistent oxidation products. UPLC-(+ESI)-QToF-MS was employed to determine major CBZ-related transformation products. Several recalcitrant hydroxy- and keto-derivatives of CBZ were tentatively identified. A Daphnia magna bioassay was used to evaluate the potential toxicity of the samples collected at different time points showing that the mixtures were highly toxic to D. magna.

Light-induced catalytic transformation of ofloxacin by solar Fenton in various water matrices at a pilot plant: mineralization and characterization of major intermediate products.

This work investigated the application of a solar driven advanced oxidation process (solar Fenton), for the degradation of the antibiotic ofloxacin (OFX) in various environmental matrices at a pilot-scale. All experiments were carried out in a compound parabolic collector pilot plant in the presence of doses of H2O2 (2.5 mg L(-1)) and at an initial Fe(2+) concentration of 2 mg L(-1). The water matrices used for the solar Fenton experiments were: demineralized water (DW), simulated natural freshwater (SW), simulated effluent from municipal wastewater treatment plant (SWW) and pre-treated real effluent from municipal wastewater treatment plant (RE) to which OFX had been spiked at 10 mg L(-1). Dissolved organic carbon removal was found to be dependent on the chemical composition of the water matrix. OFX mineralization was higher in DW (78.1%) than in SW (58.3%) at 12 mg L(-1) of H2O2 consumption, implying the complexation of iron or the scavenging of hydroxyl radicals by the inorganic ions present in SW. On the other hand, the presence of dissolved organic matter (DOM) in SWW and RE, led to lower mineralization per dose of H2O2 compared to DW and SW. The major transformation products (TPs) formed during the solar Fenton treatment of OFX, were elucidated using liquid chromatography-time of flight-mass spectrometry (LC-ToF-MS). The transformation of OFX proceeded through a defluorination reaction, accompanied by some degree of piperazine and quinolone substituent transformation while a hydroxylation mechanism occurred by attack of the hydroxyl radicals generated during the process leading to the formation of TPs in all the water matrices, seven of which were tentatively identified. The results obtained from the toxicity bioassays indicated that the toxicity originates from the DOM present in RE and its oxidation products formed during the photocatalytic treatment and not from the TPs resulted from the oxidation of OFX.

Investigating the fate of iodinated X-ray contrast media iohexol and diatrizoate during microbial degradation in an MBBR system treating urban wastewater.

The capability of a moving bed biofilm reactor (MBBR) to remove the iodinated contrast media (ICM) iohexol (IOX) and diatrizoate (DTZ) from municipal wastewater was studied. A selected number of clones of microorganisms present in the biofilm were identified. Biotransformation products were tentatively identified and the toxicity of the treated effluent was assessed. Microbial samples were DNA-sequenced and subjected to phylogenetic analysis in order to confirm the identity of the microorganisms present and determine the microbial diversity. The analysis demonstrated that the wastewater was populated by a bacterial consortium related to different members of Proteobacteria, Firmicutes, and Nitrisporae. The optimum removal values of the ICM achieved were 79 % for IOX and 73 % for DTZ, whereas 13 biotransformation products for IOX and 14 for DTZ were identified. Their determination was performed using ultra-performance liquid chromatography-tandem mass spectrometry. The toxicity of the treated effluent tested to Daphnia magna showed no statistical difference compared to that without the addition of the two ICM. The MBBR was proven to be a technology able to remove a significant percentage of the two ICM from urban wastewater without the formation of toxic biodegradation products. A large number of biotransformation products was found to be formed. Even though the amount of clones sequenced in this study does not reveal the entire bacterial diversity present, it provides an indication of the predominating phylotypes inhabiting the study site.

Superiority of solar Fenton oxidation over TiO2 photocatalysis for the degradation of trimethoprim in secondary treated effluents.

The overall aim of this work was to examine the degradation of trimethoprim (TMP), which is an antibacterial agent, during the application of two advanced oxidation process (AOP) systems in secondary treated domestic effluents. The homogeneous solar Fenton process (hv/Fe(2+)/H2O2) and heterogeneous photocatalysis with titanium dioxide (TiO2) suspensions were tested. It was found that the degradation of TMP depends on several parameters such as the amount of iron salt and H2O2, concentration of TiO2, pH of solution, solar irradiation, temperature and initial substrate concentration. The optimum dosages of Fe(2+) and H2O2 for homogeneous ([Fe(2+)] = 5 mg L(-1), [H2O2] = 3.062 mmol L(-1)) and TiO2 ([TiO2] = 3 g L(-1)) for heterogeneous photocatalysis were established. The study indicated that the degradation of TMP during the solar Fenton process is described by a pseudo-first-order reaction and the substrate degradation during the heterogeneous photocatalysis by the Langmuir-Hinshelwood kinetics. The toxicity of the treated samples was evaluated using a Daphnia magna bioassay and was finally decreased by both processes. The results indicated that solar Fenton is more effective than the solar TiO2 process, yielding complete degradation of the examined substrate within 30 min of illumination and dissolved organic carbon (DOC) reduction of about 44% whereas the respective values for the TiO2 process were ∼70% degradation of TMP within 120 min of treatment and 13% DOC removal.

Chronic ecotoxic effects to Pseudomonas putida and Vibrio fischeri, and cytostatic and genotoxic effects to the hepatoma cell line (HepG2) of ofloxacin photo(cata)lytically treated solutions.

Ofloxacin (OFL), a broad-spectrum and widespread-used photolabile fluoroquinolone, is frequently found in treated wastewaters, aquatic and terrestrial ecosystems leading to increasing concern during the past decades regarding its effects to the environment and human health. The elimination of OFL and other xenobiotics by the application of advanced oxidation processes using photolytic (PL) and photocatalytic (PC) treatments seems promising. However, an integrated assessment scheme is needed, in which, not only the removal of the parent compound, but also the effects of the photo-transformation products (PTPs) are investigated. For this purpose, in the present study, a chronic ecotoxic assessment using representative bacteria of marine and terrestrial ecosystems and a cytostatic and genotoxic evaluation using hepatoma cell line were performed. PL and PC treatments of OFL were applied using UV radiation. The photo-transformation of OFL during the treatments was monitored by DOC measurements and UPLC-MS/MS analysis. The chronic ecotoxicity of OFL and treated samples was evaluated using Pseudomonas putida and Vibrio fischeri; whereas the cytostasis and genotoxicity were estimated by the cytokinesis-block micronucleus assay (CBMN). The main results suggest that photo-transformation of OFL took place during these treatments since the concentration of OFL decreased when the irradiation time increased, as quantified by UPLC-MS/MS analysis, and this was not coupled with an analogous DOC removal. Furthermore, nine compounds were identified as probable PTPs formed through piperazinyl dealkylation and decarboxylation. The ecotoxicity of treated solutions to the bacteria studied decreased while the cytostasis to the hepatoma cell line remained at low levels during both treatments. However, the genotoxicity to the hepatoma cell line demonstrated a different pattern in which treated samples induced a greater number of MNi for the 4-16 min of irradiation (p<0.05) during both treatments. After 64 min of irradiation, the effects decreased to non genotoxic levels (p<0.05). These findings suggest that UV radiation for various treatment processes (catalytic or not), such as disinfection, may create genotoxic by-products. Therefore, in relevant technical applications, the residence time during treatment should receive special attention.

Biodegradation potential of ofloxacin and its resulting transformation products during photolytic and photocatalytic treatment.

The release of pharmaceuticals in the environment, as parent compounds, metabolites and transformation products, and the consequent risks posed to living organisms due to the unintended exposure of the latter to these chemicals are nowadays of increasing scientific concern. The development of advanced oxidation processes able to degrade these substances is in the core of the current research objectives, the main target being the removal of these compounds from wastewaters. Often the focus is on the removal of the parent compound only. However, these processes can form transformation products. Knowledge on the risk related to such transformation products is scarce. Among others, knowledge on their toxic effects and their biodegradability is of importance not only when they are present in the environment but also for the assessment of the advanced oxidation processes' efficiency applied for their degradation. Photolytic (UV irradiation) and photocatalytic treatment (UV irradiation in the presence of TiO(2)) of the fluoroquinolone ofloxacin were applied, and the biodegradability of the formed products was investigated using the Closed Bottle test (OECD 301 D). Various transformation products, formed both during the photo(cata)lytic treatment and the Closed Bottle test, were identified using chromatographic analysis with an ultra high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) system. The transformation products formed during the phototreatments were found to be non-readily biodegradable as the biodegradation percentages were close to zero. The persistence of the various photo(cata)lytic transformation products during the Closed Bottle test may be attributed to the fluorine present in all the transformation products formed. The transformation products identified suggest that two transformation routes were present: decarboxylation and opening of the piperazinyl ring. Interestingly, it was observed that in the presence of a readily biodegradable carbon source (sodium acetate), the biodegradation percentage increased drastically for some of the photolytically treated samples. This was not the case for the photocatalytically treated samples, in which also mineralization of the parent compound was achieved faster. Further research is needed, however, in order to increase the understanding of the conditions that may lead to less potent and persistent substances during the application of such engineered or natural processes.

Development and validation of a UPLC-MS/MS method for studying the degradation kinetics of ofloxacin and trimethoprim during the application of solar Fenton process in secondary treated sewage.

In this work, a sensitive and highly selective method was developed and validated to study the degradation of two antibiotic compounds (ofloxacin (OFX) and trimethoprim (TMP)), spiked in secondary treated domestic effluents, by the solar Fenton process. Three different chromatographic columns were tested on a ultra performance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS) instrument working in the electrospray ionization (ESI) mode with twelve combinations of eluting solvents. Samples were enriched prior to the analysis by solid phase extraction using the hydrophilic-lipophilic balanced (HLB) reversed phase polymeric sorbent. The method was optimized and showed very good performance characteristics and was successfully applied to study the degradation kinetics of the selected antibiotics during the solar Fenton process applied. The degradation was found to follow a pseudo first-order kinetics for both compounds at initial concentration of 100 µg L(-1) with k = 0.0345 min(-1) for OFX and k = 0.0768 min(-1) for TMP, whereas the complete removal was achieved after 120 min of treatment for both compounds.

Solar photo-Fenton process on the abatement of antibiotics at a pilot scale: Degradation kinetics, ecotoxicity and phytotoxicity assessment and removal of antibiotic resistant enterococci.

This work investigated the application of a solar driven advanced oxidation process (solar photo-Fenton), for the degradation of antibiotics at low concentration level (µg L(-1)) in secondary treated domestic effluents at a pilot-scale. The examined antibiotics were ofloxacin (OFX) and trimethoprim (TMP). A compound parabolic collector (CPC) pilot plant was used for the photocatalytic experiments. The process was mainly evaluated by a fast and reliable analytical method based on a UPLC-MS/MS system. Solar photo-Fenton process using low iron and hydrogen peroxide doses ([Fe(2+)](0) = 5 mg L(-1); [H(2)O(2)](0) = 75 mg L(-1)) was proved to be an efficient method for the elimination of these compounds with relatively high degradation rates. The photocatalytic degradation of OFX and TMP with the solar photo-Fenton process followed apparent first-order kinetics. A modification of the first-order kinetic expression was proposed and has been successfully used to explain the degradation kinetics of the compounds during the solar photo-Fenton treatment. The results demonstrated the capacity of the applied advanced process to reduce the initial wastewater toxicity against the examined plant species (Sorghum saccharatum, Lepidium sativum, Sinapis alba) and the water flea Daphnia magna. The phytotoxicity of the treated samples, expressed as root growth inhibition, was higher compared to that observed on the inhibition of seed germination. Enterococci, including those resistant to OFX and TMP, were completely eliminated at the end of the treatment. The total cost of the full scale unit for the treatment of 150 m(3) day(-1) of secondary wastewater effluent was found to be 0.85 € m(-3).

Solar photocatalytic treatment of trimethoprim in four environmental matrices at a pilot scale: transformation products and ecotoxicity evaluation.

The pilot-scale solar degradation of trimethoprim (TMP) in different water matrices (demineralized water: DW, simulated natural freshwater: SW; simulated wastewater: SWW; and real effluent: RE) was investigated in this study. DOC removal was lower in the case of SW compared to DW, which can be attributed to the presence of inorganic anions which may act as scavengers of the HO·. Furthermore, the presence of organic carbon and higher salt content in SWW and RE led to lower mineralization per dose of hydrogen peroxide compared to DW and SW. Toxicity assays in SWW and RE were also performed indicating that toxicity is attributed to the compounds present in RE and their by-products formed during solar Fenton treatment and not to the intermediates formed by the oxidation of TMP. A large number of compounds generated by the photocatalytic transformation of TMP were identified by UPLC-QToF/MS. The degradation pathway revealed differences among the four matrices; however hydroxylation, demethylation and cleavage reactions were observed in all matrices. To the best of our knowledge this is the first time that TMP degradation products have been identified by adopting a solar Fenton process at a pilot-scale set-up, using four different aqueous matrices.

Solar Fenton and solar TiO2 catalytic treatment of ofloxacin in secondary treated effluents: evaluation of operational and kinetic parameters.

Two different technical approaches based on advanced oxidation processes (AOPs), solar Fenton homogeneous photocatalysis (hv/Fe(2+)/H(2)O(2)) and heterogeneous photocatalysis with titanium dioxide (TiO(2)) suspensions were studied for the chemical degradation of the fluoroquinolone ofloxacin in secondary treated effluents. A bench-scale solar simulator in combination with an appropriate photochemical batch reactor was used to evaluate and select the optimal oxidation conditions of ofloxacin spiked in secondary treated domestic effluents. The concentration profile of the examined substrate during degradation was determined by UV/Vis spectrophotometry. Mineralization was monitored by measuring the dissolved organic carbon (DOC). The concentrations of Fe(2+) and H(2)O(2) were the key factors for the solar Fenton process, while the most important parameter of the heterogeneous photocatalysis was proved to be the catalyst loading. Kinetic analyses indicated that the photodegradation of ofloxacin can be described by a pseudo-first-order reaction. The rate constant (k) for the solar Fenton process was determined at different Fe(2+) and H(2)O(2) concentrations whereas the Langmuir-Hinshelwood (LH) kinetic expression was used to assess the kinetics of the heterogeneous photocatalytic process. The conversion of ofloxacin depends on several parameters based on the various experimental conditions, which were investigated. A Daphnia magna bioassay was used to evaluate the potential toxicity of the parent compound and its photo-oxidation by-products in different stages of oxidation. In the present study solar Fenton has been demonstrated to be more effective than the solar TiO(2) process, yielding complete degradation of the examined substrate and DOC reduction of about 50% in 30 min of the photocatalytic treatment.

Sonochemical degradation of ofloxacin in aqueous solutions.

The use of low frequency (20 kHz), high energy ultrasound for the degradation of the antibiotic ofloxacin in water was investigated. Experiments were performed with a horn-type ultrasound generator at varying applied power densities (130-640 W/L), drug concentrations (5-20 mg/L), hydrogen peroxide concentrations (0-100 mM) and sparging gases (air, oxygen, nitrogen and argon). In general, conversion (which was assessed following sample absorbance at 288 nm) increased with increasing ultrasound energy and peroxide concentration and decreasing initial drug concentration. Moreover, reactions under an argon atmosphere were faster than with diatomic gases, possibly due to argon's physical properties (e.g. solubility, thermal conductivity and specific heat ratio) favoring sonochemical activity. Overall, low to moderate levels of ofloxacin degradation were achieved (i.e. it never exceeded 50%), thus indicating that radical reactions in the liquid bulk rather than thermal reactions in the vicinity of the cavitation bubble are responsible for ofloxacin degradation.

Drugs degrading photocatalytically: Kinetics and mechanisms of ofloxacin and atenolol removal on titania suspensions.

The conversion of the antibiotic ofloxacin and the beta-blocker atenolol by means of TiO(2) photocatalysis was investigated. Irradiation was provided by a UVA lamp at 3.37x10(-6)einstein/s photon flux, while emphasis was given on the effect of catalyst type and loading (50-1500mg/L), initial substrate concentration (5-20mg/L), initial pH (3-10) and the effect of H(2)O(2) (0.07-1.4mM) as an additional oxidant on substrate conversion and mineralization in various matrices (i.e. pure water, groundwater and treated municipal effluent). Conversion was assessed measuring sample absorbance at 288 and 224nm for ofloxacin and atenolol, respectively, while mineralization measuring the dissolved organic carbon. Degussa P25 TiO(2) was found to be more active than other TiO(2) samples for either substrate degradation, with ofloxacin being more reactive than atenolol. Conversion generally increased with increasing catalyst loading, decreasing initial substrate concentration and adding H(2)O(2), while the effect of solution pH was substrate-specific. Reaction rates, following a Langmuir-Hinshelwood kinetic expression, were maximized at a catalyst to substrate concentration ratio (w/w) of 50 and 15 for ofloxacin and atenolol, respectively, while higher ratios led to reduced efficiency. Likewise, high concentrations of H(2)O(2) had an adverse effect on reaction, presumably due to excessive oxidant scavenging radicals and other reactive species. The ecotoxicity of ofloxacin and atenolol to freshwater species Daphnia magna was found to increase with increasing substrate concentration (1-10mg/L) and exposure time (24-48h), with atenolol being more toxic than ofloxacin. Photocatalytic treatment eliminated nearly completely toxicity and this was more pronounced for atenolol.

Rapid and sensitive high-performance liquid chromatographic determination of four cephalosporin antibiotics in pharmaceuticals and body fluids.

A rapid, accurate and sensitive method has been developed and validated for the quantitative simultaneous determination of four cephalosporins, cephalexin and cefadroxil (first-generation), cefaclor (second-generation) and cefataxim (third-generation), in pharmaceuticals as well as in human blood serum and urine. A Spherisorb ODS-2 250 x 4-mm, 5-microm analytical column was used with an eluting system consisting of a mixture of acetate buffer (pH 4.0)-CH(3)OH 78-22% (v/v) at a flow-rate 1.2 ml/min. Detection was performed with a variable wavelength UV-Vis detector at 265 nm resulting in limit of detection of 0.2 ng for cefadroxil and cephalexin, but only 0.1 ng for cefotaxime and cefaclor per 20-microl injection. Hydrochlorothiazide (HCT) (6-chloro-3,4-dihydro-7 sulfanyl-2H-1,2,4-benzothiadiazine-1-1-dioxide) was used as internal standard at a concentration of 2 ng/microl. A rectilinear relationship was observed up to 8, 5, 12 and 35 ng/microl for cefadroxil, cefotaxime, cefaclor, cephalexin, respectively. Analysis time was less than 7 min. The statistical evaluation of the method was examined by means of within-day repeatability (n=8) and day-to-day precision (n=9) and was found to be satisfactory with high accuracy and precision. The method was applied to the determination of the cephalosporins in commercial pharmaceuticals and in biological fluids: human blood serum after solid-phase extraction and urine simply after filtration and dilution. Recovery of analytes in spiked samples was in the range from 76.3 to 112.0%, over the range of 1-8 ng/microl.