RESUMO
6PPD, a tire rubber antioxidant, poses substantial ecological risks because it can form a highly toxic quinone transformation product (TP), 6PPD-quinone (6PPDQ), during exposure to gas-phase ozone. Important data gaps exist regarding the structures, reaction mechanisms, and environmental occurrence of TPs from 6PPD ozonation. To address these data gaps, gas-phase ozonation of 6PPD was conducted over 24-168 h and ozonation TPs were characterized using high-resolution mass spectrometry. The probable structures were proposed for 23 TPs with 5 subsequently standard-verified. Consistent with prior findings, 6PPDQ (C18H22N2O2) was one of the major TPs in 6PPD ozonation (â¼1 to 19% yield). Notably, 6PPDQ was not observed during ozonation of 6QDI (N-(1,3-dimethylbutyl)-N'-phenyl-p-quinonediimine), indicating that 6PPDQ formation does not proceed through 6QDI or associated 6QDI TPs. Other major 6PPD TPs included multiple C18H22N2O and C18H22N2O2 isomers, with presumptive N-oxide, N,N'-dioxide, and orthoquinone structures. Standard-verified TPs were quantified in roadway-impacted environmental samples, with total concentrations of 130 ± 3.2 µg/g in methanol extracts of tire tread wear particles (TWPs), 34 ± 4 µg/g-TWP in aqueous TWP leachates, 2700 ± 1500 ng/L in roadway runoff, and 1900 ± 1200 ng/L in roadway-impacted creeks. These data demonstrate that 6PPD TPs are likely an important and ubiquitous class of contaminants in roadway-impacted environments.
Assuntos
Antioxidantes , Benzoquinonas , Fenilenodiaminas , Borracha , Antioxidantes/química , Ozônio/química , Borracha/química , Água/química , Fenilenodiaminas/química , Benzoquinonas/químicaRESUMO
Recently, roadway releases of N,N'-substituted p-phenylenediamine (PPD) antioxidants and their transformation products (TPs) received significant attention due to the highly toxic 6PPD-quinone. However, the occurrence of PPDs and TPs in recycled tire rubber products remains uncharacterized. Here, we analyzed tire wear particles (TWPs), recycled rubber doormats, and turf-field crumb rubbers for seven PPD antioxidants, five PPD-quinones (PPDQs), and five other 6PPD TPs using liquid chromatography-tandem mass spectrometry. PPD antioxidants, PPDQs, and other TPs were present in all samples with chemical profiles dominated by 6PPD, DTPD, DPPD, and their corresponding PPDQs. Interestingly, the individual [PPDQ]/[PPD] and [TP]/[PPD] ratios significantly increased as total concentrations of the PPD-derived chemical decreased, indicating that TPs (including PPDQs) dominated the PPD-derived compounds with increased environmental weathering. Furthermore, we quantified 15 other industrial rubber additives (including bonding agents, vulcanization accelerators, benzotriazole and benzothiazole derivatives, and diphenylamine antioxidants), observing that PPD-derived chemical concentrations were 0.5-6 times higher than these often-studied additives. We also screened various other elastomeric consumer products, consistently detecting PPD-derived compounds in lab stoppers, sneaker soles, and rubber garden hose samples. These data emphasize that PPD antioxidants, PPDQs, and related TPs are important, previously overlooked contaminant classes in tire rubbers and elastomeric consumer products.
Assuntos
Antioxidantes , Benzoquinonas , Fenilenodiaminas , Borracha , Antioxidantes/análise , Antioxidantes/química , Antioxidantes/classificação , Fenilenodiaminas/análise , Fenilenodiaminas/química , Fenilenodiaminas/classificação , Borracha/química , Benzoquinonas/análise , Benzoquinonas/química , Benzoquinonas/classificação , Espectrometria de Massa com Cromatografia Líquida , Espectrometria de Massas em TandemRESUMO
Free chlorine disinfection is widely applied to inactivate viruses by reacting with their biomolecules, which include nucleic acids, proteins, and lipids. Knowing the reactivities of viral genomes with free chlorine and the protection that encapsidation provides would ultimately help predict virus susceptibility to the disinfectant. The relative reactivities of different viral genome types and the impact of viral higher order structure with free chlorine are poorly characterized. Here, we studied the reactivity of viral genomes representing four genome types from virus particles with diverse structures, namely, (+)ssRNA (MS2), dsRNA (φ6), ssDNA (φX174), and dsDNA (T3) with free chlorine. We compared the reactivities of these viral nucleic acids when they were suspended in phosphate buffer solutions (naked forms) and when they were in the native virus particles (encapsidated forms). The reactivities of nucleic acids were tracked by polymerase chain reaction (PCR)-based assays. The naked dsDNA of T3 was the least reactive with free chlorine, with an average second order rate constant normalized by the number of bases in the measured regions (in M-1 s-1 b-1) that was 34×, 65×, and 189× lower than those of the dsRNA of φ6, ssRNA of MS2, and ssDNA of φX174, respectively. Moreover, different regions in the ssRNA genome of MS2 and the dsRNA genome of φ6 exhibited statistically different reaction kinetics. The genomes within virus particles reacted slower than the naked genomes overall, but the extent of these differences varied among the four viruses. The results on viral nucleic acid reactivity help explain different susceptibilities of viruses to inactivation by free chlorine and also provide a valuable comparison of the susceptibilities of different nucleic acids to oxidants.
Assuntos
Ácidos Nucleicos , Vírus , Cloro/farmacologia , Desinfecção/métodos , Inativação de VírusRESUMO
This study investigated antibiotic resistance gene (ARG) degradation kinetics in wastewaters during bench- and full-scale treatment with UV light and chlorineâwith the latter maintained as free available chlorine (FAC) in low-ammonia wastewater and converted into monochloramine (NH2Cl) in high-ammonia wastewater. Twenty-three 142-1509 bp segments (i.e., amplicons) of seven ARGs (blt, mecA, vanA, tet(A), ampC, blaNDM, blaKPC) and the 16S rRNA gene from antibiotic resistant bacteria (ARB) strains Bacillus subtilis, Staphylococcus aureus, Enterococcus faecium, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae were monitored as disinfection targets by qPCR. Rate constants for ARG and 16S rRNA gene amplicon degradation by UV, FAC, and NH2Cl were measured in phosphate buffer and used to expand and validate several recently developed approaches to predict DNA segment degradation rate constants based solely on their nucleotide contents, which were then applied to model ARG degradation during bench-scale treatment in buffer and wastewater matrixes. Kinetics of extracellular and intracellular ARG degradation by UV and FAC were well predicted up to â¼1-2-log10 elimination, although with decreasing accuracy at higher levels for intracellular genes, while NH2Cl yielded minimal degradation under all conditions (agreeing with predictions). ARB inactivation kinetics varied substantially across strains, with intracellular gene degradation lagging cell inactivation in each case. ARG degradation levels observed during full-scale disinfection at two wastewater treatment facilities were consistent with bench-scale measurements and predictions, where UV provided â¼1-log10 ARG degradation, and chlorination of high-ammonia wastewater (dominated by NH2Cl) yielded minimal ARG degradation.
Assuntos
Cloro , Purificação da Água , Águas Residuárias/microbiologia , Desinfecção , Raios Ultravioleta , RNA Ribossômico 16S , Nucleotídeos , Amônia , Antagonistas de Receptores de Angiotensina , Inibidores da Enzima Conversora de Angiotensina , Escherichia coli , Resistência Microbiana a Medicamentos/genética , Antibacterianos/farmacologiaRESUMO
Degradation kinetics of antibiotic resistance genes (ARGs) by free available chlorine (FAC), ozone (O3), and UV254 light (UV) were investigated in phosphate buffered solutions at pH 7 using a chromosomal ARG (mecA) of methicillin-resistant Staphylococcus aureus (MRSA). For FAC, the degradation rates of extracellular mecA (extra-mecA) were accelerated with increasing FAC exposure, which could be explained by a two-step FAC reaction model. The degradation of extra-mecA by O3 followed second-order reaction kinetics. The degradation of extra-mecA by UV exhibited tailing kinetics, which could be described by a newly proposed kinetic model considering cyclobutane pyrimidine dimer (CPD) formation, its photoreversal, and irreversible (6-4) photoproduct formation. Measured rate constants for extra-mecA increased linearly with amplicon length for FAC and O3, or with number of intrastrand pyrimidine doublets for UV, which enabled prediction of degradation rate constants of extra-mecA amplicons based on sequence length and/or composition. In comparison to those of extra-mecA, the observed degradation rates of intracellular mecA (intra-mecA) were faster for FAC and O3 at low oxidant exposures but significantly slower at high exposures for FAC and UV. Differences in observed extra- and intracellular kinetics could be due to decreased DNA recovery efficiency and/or the presence of MRSA aggregates protected from disinfectants.
Assuntos
Staphylococcus aureus Resistente à Meticilina , Ozônio , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Cloro , Desinfecção , Resistência Microbiana a Medicamentos , Cinética , Staphylococcus aureus Resistente à Meticilina/genética , Raios Ultravioleta , ÁguaRESUMO
This work investigated degradation (measured by qPCR) and biological deactivation (measured by culture-based natural transformation) of extra- and intracellular antibiotic resistance genes (eARGs and iARGs) by free available chlorine (FAC), NH2Cl, O3, ClO2, and UV light (254 nm), and of eARGs by â¢OH, using a chromosomal ARG ( blt) of multidrug-resistant Bacillus subtilis 1A189. Rate constants for degradation of four 266-1017 bp amplicons adjacent to or encompassing the acfA mutation enabling blt overexpression increased in proportion to #AT+GC bps/amplicon, or in proportion to #5'-GG-3' or 5'-TT-3' doublets/amplicon, with respective values ranging from 0.59 to 2.3 (×1011 M-1 s-1) for â¢OH, 1.8-6.9 (×104 M-1 s-1) for O3, 3.9-9.2 (×103 M-1 s-1) for FAC, 0.35-1.2(×101 M-1 s-1) for ClO2, and 2.0-8.8 (×10-2 cm2/mJ) for UV at pH 7, and from 1.7-4.4 M-1 s-1 for NH2Cl at pH 8. For FAC, NH2Cl, O3, ClO2, and UV, ARG deactivation paralleled degradation of amplicons approximating a â¼800-1000 bp acfA-flanking sequence required for natural transformation in B. subtilis, whereas deactivation outpaced degradation for â¢OH. At practical disinfectant exposures, eARGs and iARGs were ≥90% degraded/deactivated by FAC, O3, and UV, but recalcitrant to NH2Cl and ClO2. iARG degradation/ deactivation always lagged cell inactivation. These findings provide a quantitative framework for evaluating ARG fate during disinfection/oxidation, and support using qPCR as a proxy for tracking ARG deactivation under carefully selected circumstances.
Assuntos
Ozônio , Purificação da Água , Cloraminas , Cloro , Compostos Clorados , Radical Hidroxila , Óxidos , Raios UltravioletaRESUMO
Oxidation of ß-lactam antibiotics by aqueous ferrate(VI) was investigated to determine reaction kinetics, reaction sites, antibacterial activity changes, and transformation products. Apparent second-order rate constants (kapp) were determined in the pH range 6.0-9.5 for the reaction of ferrate(VI) with penicillins (amoxicillin, ampicillin, cloxacillin, and penicillin G), a cephalosporin (cephalexin), and several model compounds. Ferrate(VI) shows an appreciable reactivity toward the selected ß-lactams (kapp for pH 7 = 110-770 M(-1) s(-1)). The pH-dependent kapp could be well explained by considering species-specific reactions between ferrate(VI) and the ß-lactams (with reactions occurring at thioether, amine, and/or phenol groups). On the basis of the kinetic results, the thioether is the main reaction site for cloxacillin and penicillin G. In addition to the thioether, the amine is a reaction site for ampicillin and cephalexin, and amine and phenol are reaction sites for amoxicillin. HPLC/MS analysis showed that the thioether of ß-lactams was transformed to stereoisomeric (R)- and (S)-sulfoxides and then to a sulfone. Quantitative microbiological assay of ferrate(VI)-treated ß-lactam solutions indicated that transformation products resulting from the oxidation of cephalexin exhibited diminished, but non-negligible residual activity (i.e., â¼24% as potent as the parent compound). For the other ß-lactams, the transformation products showed much lower (<5%) antibacterial potencies compared to the parent compounds. Overall, ferrate(VI) oxidation appears to be effective as a means of lowering the antibacterial activities of ß-lactams, although alternative approaches may be necessary to achieve complete elimination of cephalosporin activities.
Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Ferro/química , beta-Lactamas/química , beta-Lactamas/farmacologia , Bacillus subtilis/efeitos dos fármacos , Biotransformação/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Cinética , Testes de Sensibilidade Microbiana , Oxirredução , Fenóis/química , Águas Residuárias/químicaRESUMO
Reactions of dissolved organic matter (DOM) with photochemically generated reactive halogen species (RHS) may represent an important natural source of organohalogens within surface seawaters. However, investigation of such processes has been limited by difficulties in quantifying low dissolved organohalogen concentrations in the presence of background inorganic halides. In this work, sequential solid phase extraction (SPE) and silver-form cation exchange filtration were utilized to desalt and preconcentrate seawater DOM prior to nonspecific organohalogen analysis by ICP-MS. Using this approach, native organobromine and organoiodine contents were found to range from 3.2-6.4 × 10(-4) mol Br/mol C and 1.1-3.8 × 10(-4) mol I/mol C (or 19-160 nmol Br L(-1) and 6-36 nmol I L(-1)) within a wide variety of natural seawater samples, compared with 0.6-1.2 × 10(-4) mol Br/mol C and 0.6-1.1 × 10(-5) mol I/mol C in terrestrial natural organic matter (NOM) isolates. Together with a chemical probe method specific for RHS, the SPE+ICP-MS approach was also employed to demonstrate formation of nanomolar levels of organobromine and organoiodine during simulated and natural solar irradiation of DOM in artificial and natural seawaters. In a typical experiment, the organobromine content of 2.1 × 10(-4) mol C L(-1) (2.5 mg C L(-1)) of Suwannee River NOM in artificial seawater increased by 69% (from 5.9 × 10(-5) to 1.0 × 10(-4) mol Br/mol C) during exposure to 24 h of simulated sunlight. Increasing I(-) concentrations (up to 2.0 × 10(-7) mol L(-1)) promoted increases of up to 460% in organoiodine content (from 8.5 × 10(-6) to 4.8 × 10(-5) mol I/mol C) at the expense of organobromine formation under the same conditions. The results reported herein suggest that sunlight-driven reactions of RHS with DOM may play a significant role in marine bromine and iodine cycling.
Assuntos
Bromo/análise , Halogenação/efeitos da radiação , Hidrocarbonetos Halogenados/análise , Iodo/análise , Processos Fotoquímicos/efeitos da radiação , Água do Mar/química , Luz Solar , Meio Ambiente , Radicais Livres/química , Substâncias Húmicas/análise , Hidrocarbonetos Halogenados/isolamento & purificação , Pirazóis/química , Rios/químicaRESUMO
Combinations of UV with oxidants can initiate advanced oxidation processes (AOPs) and enhance bacterial inactivation. However, the effectiveness and mechanisms of UV-AOPs in damaging nucleic acids (e.g., antibiotic resistance genes (ARGs)) and cell integrity represent a knowledge gap. This study comprehensively compared ARG degradation and cell membrane damage under three different UV-AOPs. The extracellular ARG (eARG) removal efficiency followed the order of UV/chlorine > UV/H2O2 > UV/peracetic acid (PAA). Hydroxyl radical (â¢OH) and reactive chlorine species (RCS) largely contributed to eARG removal, while organic radicals made a minor contribution. For intracellular ARGs (iARGs), UV/H2O2 did not remove better than UV alone due to the scavenging of â¢OH by cell components, whereas UV/PAA provided a modest synergism, likely due to diffusion of PAA into cells and intracellular â¢OH generation. Comparatively, UV/chlorine achieved significant synergistic iARG removal, suggesting the critical role of the RCS in resisting cellular scavenging and inactivating ARGs. Additionally, flow cytometry analysis demonstrated that membrane damage was mainly attributed to chlorine oxidation, while the impacts of radicals, H2O2, and PAA were negligible. These results provide mechanistic insights into bacterial inactivation and fate of ARGs during UV-AOPs, and shed light on the suitability of quantitative polymerase chain reaction (qPCR) and flow cytometry in assessing disinfection performance.
RESUMO
Aqueous free available chlorine (FAC) can be photolyzed by sunlight and/or artificial UV light to generate various reactive oxygen species, including HO(â¢) and O((3)P). The influence of this chemistry on inactivation of chlorine-resistant microorganisms was investigated using Bacillus subtilis endospores as model microbial agents and simulated and natural solar radiation as light sources. Irradiation of FAC solutions markedly enhanced inactivation of B. subtilis spores in 10 mM phosphate buffer; increasing inactivation rate constants by as much as 600%, shortening inactivation curve lag phase by up to 73% and lowering CTs required for 2 log10 inactivation by as much as 71% at pH 8.0 and 10 °C. Similar results were observed at pH 7.4 and 10 °C in two drinking water samples with respective DOC concentrations and alkalinities of 0.6 and 1.2 mg C/L and 81.8 and 17.1 mg/L as CaCO3. Solar radiation alone did not inactivate B. subtilis spores under the conditions investigated. A variety of experimental data indicate that the observed enhancements in spore inactivation can be attributed to the concomitant attack of spores by HO(â¢) and O3, the latter of which was found to accumulate to micromolar concentrations during simulated solar irradiation of 10 mM phosphate buffer (pH 8, 10 °C) containing [FAC]0 = 8 mg/L as Cl2.
Assuntos
Bacillus subtilis/efeitos da radiação , Cloro/efeitos da radiação , Viabilidade Microbiana/efeitos da radiação , Fotólise/efeitos da radiação , Esporos Bacterianos/efeitos da radiação , Luz Solar , Bacillus subtilis/fisiologia , Concentração de Íons de Hidrogênio , Oxidantes/química , Espécies Reativas de Oxigênio/análise , Esporos Bacterianos/fisiologia , Temperatura , Microbiologia da ÁguaRESUMO
We here report chemical characteristics relevant to the fate and transport of the recently discovered environmental toxicant 6PPD-quinone (2-((4-methylpentan-2-yl)amino)-5-(phenylamino)cyclohexa-2,5-diene-1,4-dione or "6PPDQ"). 6PPDQ is a transformation product of the tire rubber antioxidant 6PPD that is ubiquitous in roadway environments, including atmospheric particulate matter, soils, runoff, and receiving waters, after dispersal from tire rubber use and wear on roadways. The aqueous solubility and octanol-water partitioning coefficient (i.e. log KOW) for 6PPDQ were measured to be 38 ± 10 µg L-1 and 4.30 ± 0.02, respectively. Within the context of analytical measurement and laboratory processing, sorption to various laboratory materials was evaluated, indicating that glass was largely inert but loss of 6PPDQ to other materials was common. Aqueous leaching simulations from tire tread wear particles (TWPs) indicated short term release of â¼5.2 µg 6PPDQ per gram TWP over 6 h under flow-through conditions. Aqueous stability tests observed a slight-to-moderate loss of 6PPDQ over 47 days (26 ± 3% loss) for pH 5, 7 and 9. These measured physicochemical properties suggest that 6PPDQ is generally poorly soluble but fairly stable over short time periods in simple aqueous systems. 6PPDQ can also leach readily from TWPs for subsequent environmental transport, posing high potential for adverse effects in local aquatic environments.
Assuntos
Benzoquinonas , Substâncias Perigosas , Fenilenodiaminas , Borracha , Poluentes Químicos da Água , Substâncias Perigosas/química , Material Particulado/química , Água/química , Poluentes Químicos da Água/química , Fenilenodiaminas/química , Benzoquinonas/química , SolubilidadeRESUMO
Antibiotic resistance genes (ARGs), in association with antibiotic resistant bacteria (ARB), have been identified as widespread contaminants of treated drinking waters and wastewaters. As a consequence, concerns have been raised that ARB or ARG transport between aquatic compartments may enhance the spread of antibiotic resistance amongst non-resistant bacterial communities by means of horizontal gene transfer processes. Most often, discussion of horizontal gene transfer focuses on the probable role of conjugative plasmid or transposon exchange, which requires live ARB donor cells. Conventional water and wastewater disinfection processes generally provide highly effective means for mitigating the transport of live ARB; thereby minimizing risks of conjugative gene transfer. However, even if ARB present in a treated water are fully inactivated during a disinfection process, the possibility remains that intact remnants of DNA contained within the resulting cell debris could still confer resistance genotypes to downstream bacterial populations by means of natural transformation and/or transduction, which do not require live donor cells. Thus, a systematic evaluation of the capability of common disinfection technologies to ensure the destruction of bacterial DNA, in addition to pathogen inactivation, seems warranted. With that objective in mind, this review seeks to provide a concise introduction to the significance of ARB and ARG occurrence in environmental systems, coupled with a review of the role that commonly used water and wastewater disinfection processes may play in minimizing ARG transport and dissemination.
Assuntos
Bactérias/genética , Desinfecção , Resistência Microbiana a Medicamentos/genética , Eliminação de Resíduos Líquidos , Microbiologia da Água , Bactérias/efeitos dos fármacos , Transferência Genética Horizontal , Genes BacterianosRESUMO
The reaction of hypochlorous acid (HOCl) with superoxide radical (O2â¢-) - a source of hydroxyl radical (HOâ¢) and various reactive chlorine species (RCS) - was investigated as the basis for a novel non-photochemical advanced oxidation process (AOP). Moderately stable (t1/2 ~ minutes) aqueous O2â¢- stocks were prepared by several approaches at pH>12 and either (a) added directly to aqueous free available chlorine (FAC; i.e., HOCl/OCl-) at circumneutral pH, or (b) premixed with alkaline FAC and then acidified to pH 7, to degrade various organic probe compounds via in situ generated HO⢠and RCS. Radical production was optimal at [HO2â¢/O2â¢-]0/[FAC]0 ~ 2, with ~0.8 mol HO⢠formed/mol FAC consumed, and HO⢠and RCS exposures reaching ~5×10-10 and ~10-9 M×s, respectively. Similar trends were observed in natural waters and organic matter-amended phosphate buffer containing up to 5 mgC/L of dissolved organic carbon. Direct formation of oxyhalides, trihalomethanes (THMs), and haloacetic acids (HAAs), was minimal, though THM and HAA formation was moderately enhanced during post-chlorination of O2â¢-/FAC-treated solutions. This process could provide a beneficial addition to the range of available AOPs due to its high radical exposures, simplicity, rapid time-scales, potential for on-site O2â¢- generation, and widespread accessibility of FAC and other reagents.
Assuntos
Poluentes Químicos da Água , Purificação da Água , Cloro , Desinfecção , Halogenação , Radical Hidroxila , Ácido Hipocloroso , Superóxidos , Poluentes Químicos da Água/análiseRESUMO
Degradation and deactivation kinetics of an antibiotic resistance gene (ARG) by ozone (O3) and free available chlorine (FAC) were investigated in phosphate-buffered solutions at pH 7 for O3 (in the presence of tertbutanol), and pH 6.8 or 8.1 for FAC. We used a plasmid (pUC19)-encoded ampicillin resistance gene (ampR) in both extracellular (e-) and intracellular (i-) forms. The second-order rate constant (kO3) for degradation of 2686 base pair (bp) long e-pUC19 toward O3, which was determined by quantitative polymerase chain reaction assay, was calculated to be ~2 × 105 M-1s-1. The deactivation rate constants of e-pUC19 by O3 measured with various recipient E. coli strains were within a factor of 2 compared with the degradation rate constant for e-pUC19. The degradation/deactivation kinetics of i-pUC19 were similar to those of e-pUC19, indicating only a minor influence of cellular components on O3 reactivity toward i-pUC19. For FAC, the degradation and deactivation rates of e-pUC19 were decreased in the presence of tertbutanol, implying involvement of direct FAC as well as some radical (e.g., â¢OH) reactions. The degradation rates of e-ampR segments by direct FAC reaction could be explained by a previously-reported two-step sequential reaction model, in which the rate constants increased linearly with e-ampR segment length. The deactivation rate constants of e-pUC19 during exposure to FAC were variable by a factor of up to 4.3 for the different recipient strains, revealing the role of DNA repair in the observed deactivation efficiencies. The degradation/deactivation of e-pUC19 were significantly faster at pH 6.8 than at pH 8.1 owing to pH-dependent FAC speciation variation, whereas i-pUC19 kinetics exhibited much smaller dependence on pH, demonstrating intracellular plasmid DNA reactions with FAC occurred at cytoplasmic pH (~7.5). Our results are useful for predicting and/or measuring the degradation/deactivation efficiency of plasmid-encoded ARGs by water treatment with ozonation and chlorination.
Assuntos
Ozônio , Poluentes Químicos da Água , Purificação da Água , Antibacterianos/farmacologia , Cloro , Resistência Microbiana a Medicamentos/genética , Escherichia coli/genética , Cinética , Oxirredução , Plasmídeos/genéticaRESUMO
A simple and rapid method employing non-suppressed ion chromatography with electrospray ionization tandem mass spectrometry has been developed for the direct determination of trace-level haloacetic acids (HAAs) in water samples. Using 70/30 (v/v) acetonitrile/1 M aqueous methylamine as the mobile phase, three IC columns - AS16, AS18 and AS24 from Thermo-Scientific - were tested, respectively, with the AS16 column exhibiting the best overall performance with respect to resolution and retention time. To assess the effects of mobile phase composition on retention time of HAAs, the AS16 column was further tested using (i) different proportions of acetonitrile to aqueous methylamine, (ii) different proportions of acetonitrile to aqueous solution at fixed methylamine concentrations, and (iii) different concentrations of methylamine at fixed proportions of acetonitrile to aqueous solution. With a low proportion of aqueous solution, van der Waals and/or hydrogen-bonding interactions appeared to play an important role in governing HAA retention, i.e., HAAs with relatively higher apparent logKow* caused by elevated solvent sspKa exhibited longer retention times; whereas with a high proportion of aqueous solution, ionic interactions appeared to dominate retention of HAAs, with the more polarizable HAAs exhibiting longer retention times. Using 70/30 (v/v) acetonitrile/1 M aqueous methylamine, the method detection limits were in the range of 0.090-0.216 µg/L for the 11 selected chloro-, bromo- and iodoacetic acids. Finally, this method was applied to monitor HAAs yields in laboratory chlorination experiments and to determine concentrations of HAAs in tap water and wastewater effluent samples.
RESUMO
In U.S. Pacific Northwest coho salmon (Oncorhynchus kisutch), stormwater exposure annually causes unexplained acute mortality when adult salmon migrate to urban creeks to reproduce. By investigating this phenomenon, we identified a highly toxic quinone transformation product of N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD), a globally ubiquitous tire rubber antioxidant. Retrospective analysis of representative roadway runoff and stormwater-affected creeks of the U.S. West Coast indicated widespread occurrence of 6PPD-quinone (<0.3 to 19 micrograms per liter) at toxic concentrations (median lethal concentration of 0.8 ± 0.16 micrograms per liter). These results reveal unanticipated risks of 6PPD antioxidants to an aquatic species and imply toxicological relevance for dissipated tire rubber residues.
Assuntos
Antioxidantes/toxicidade , Benzoquinonas/toxicidade , Exposição Ambiental , Oncorhynchus kisutch/fisiologia , Fenilenodiaminas/toxicidade , Borracha/toxicidade , Animais , Noroeste dos Estados Unidos , Borracha/químicaRESUMO
Reactions of ozone (O3) with the beta-lactam antibiotics penicillin G (PG) and cephalexin (CP) have previously been found to yield products retaining antibacterial activities. These products are unequivocally identified here as the stereoisomeric (R)-sulfoxides of each parent molecule and characterized by a combination of chemical analysis and an antibacterial activity assay. PG-(R)-sulfoxide, which is approximately 15% as potent as PG itself, is formed in approximately 55% yield, whereas CP-(R)-sulfoxide, which is 83% as active as CP, is formed with a maximum 34% yield. PG-(R)-sulfoxide is recalcitrant toward further oxidation by O3, but readily transformed by hydroxyl radical (HO*) (kHO*,app"=7.4x10(9) M(-1) s(-1), pH 7), resulting in quantitative elimination of its antibacterial activity. In contrast, CP-(R)-sulfoxide is degraded by both O3 and HO* (kO3,app"=2.6x10(4) M(-1) s(-1) and kHO*,app"= 7.6x10(9) M(-1) s(-1), pH 7), leading to quantitative elimination of its antibacterial activity. During ozonation of a secondary municipal wastewater effluent sample (pH 8.1, CDOC=4.0 mg/L, [alkalinity]=3.6 mM as HCO3-) spiked with [PG]0=1 microM, PG-(R)-sulfoxide yields did not exceed 0.15 microM for O3 doses up to 100 microM (4.8 mg/L), but reached 0.47 microM with 10-mM t-BuOH added as a HO* scavenger. In contrast, CP-(R)-sulfoxide yields did not exceed 0.1 microM for the same wastewater spiked with [CP]0=1 microM in either the presence or absence of t-BuOH, indicating that CP-(R)-sulfoxide transformation is governed primarily by direct reaction with O3. These findings suggest that, for a given degree of parent compound transformation, PG-(R)-sulfoxide yields would likely be greatest during ozonation of wastewaters characterized by low O3 demands and high HO* scavenging rates, whereas CP-(R)-sulfoxide yields would be less matrix-dependent. In general, complete deactivation of penicillins during wastewater treatment will likely require higher O3 exposures than necessary for deactivation of cephalosporins.
Assuntos
Antibacterianos/química , Cefalexina/química , Ozônio/química , Penicilina G/química , Biotransformação , Radical Hidroxila/química , Oxirredução , Eliminação de Resíduos Líquidos/métodosRESUMO
This study investigated the degradation and deactivation of an extracellular ampicillin resistance gene (ampR) encoded in plasmid pUC19 during exposure to UV254, â¢OH (generated by UV>290/H2O2), and combined exposure to UV254 and â¢OH (and/or SO4â¢-) using UV254/H2O2 and UV254/S2O82-. The degradation rates of ampR measured by quantitative polymerase chain reaction increased with increasing target amplicon length (192-851 bps). The rate constants for the degradation of pUC19 (2686 bps) were calculated as 0.26 cm2/mJ for UV254 and 1.5 × 1011 M-1s-1 for â¢OH, based on the degradation rates of ampR amplicons and assuming an equal sensitivity of DNA damage across the entire plasmid. DNA repair-proficient Escherichia coli (E. coli) AB1157 strain (wild-type) and its repair-deficient mutants including AB1886 (uvrA-), AB2463 (recA-), AB2480 (uvrA-, recA-), and DH5α (recA-, endA-) were applied as recipient cells in gene transformation assays. Results suggested that the elimination efficiency of transforming activity during UV254 and â¢OH exposure was dependent on the type of DNA repair genes in recipient E. coli strains. Losses of transforming activity were slower than the degradation of pUC19 by a factor of up to â¼5 (for E. coli DH5α), highlighting the importance of DNA repair in recipient cells. The degradation rates of ampR amplicons were much larger (by a factor of â¼4) in UV254/H2O2 and UV254/S2O82- than UV254 direct photolysis, indicating the significant contribution of â¢OH and SO4â¢- to the gene degradation. Not only UV254 and SO4â¢-, but also â¢OH contributed to the degradation of ampR during UV254/S2O82-, which was attributed to the conversion of SO4â¢- to â¢OH and a 10-fold larger reactivity of â¢OH towards ampR as compared to SO4â¢-. However, the enhanced gene degradation by radicals did not lead to a faster elimination of gene transforming activity during UV254/H2O2 and UV254/S2O82-, suggesting that UV254- and radical-induced DNA damage were not additive in their contributions to losses of gene transforming activity. Wastewater effluent organic matter (EfOM) accelerated the degradation of ampR during UV254 irradiation by means of reactive species production through indirect photolysis reactions, whereas EfOM mainly acted as a radical scavenger during UV254/H2O2 and UV254/S2O82- treatments.
Assuntos
Antibacterianos , Poluentes Químicos da Água , Resistência Microbiana a Medicamentos , Escherichia coli/genética , Peróxido de Hidrogênio , Plasmídeos , Raios UltravioletaRESUMO
Solar irradiation of chlorine-containing waters enhances inactivation of chlorine-resistant pathogens (e.g., Cryptosporidium oocysts), through in situ formation of ozone, hydroxyl radical, and other reactive species during photolysis of free available chlorine (FAC) at UVB-UVA wavelengths of solar light (290-400â¯nm). However, corresponding effects on regulated disinfection byproduct (DBP) formation and associated dissolved organic matter (DOM) properties remain unclear. In this work, when compared to dark chlorination, sunlight-driven FAC photolysis over a range of conditions was found to yield higher DBP levels, depletion of DOM chromophores and fluorophores, preferential removal of phenolic groups versus carboxylic acid groups, and degradation of larger humic substances to smaller molecular weight compounds. Control experiments showed that increased DBP levels were not due to direct DOM photolysis and subsequent dark reactions with FAC, but to co-exposure of DOM to FAC and reactive species (e.g., O3, HOâ¢, Clâ¢, Cl2â¢-, ClOâ¢) generated by FAC photolysis. Because solar chlorine photolysis can enable inactivation of chlorine-resistant pathogens at far lower CTFAC values than chlorination alone, the increases in DBP formation inherent to this approach can likely be offset to some extent by the ability to operate at significantly decreased CTFAC. Nonetheless, these findings demonstrate that applications of solar chlorine photolysis will require careful attention to potential impacts on DBP formation.