Responses of nitrification performance, triclosan resistome and diversity of microbes to continuous triclosan stress in activated sludge system

Triclosan(TCS) is commonly found in wastewater treatment plants,which often affects biological treatment processes.The responses of nitrification,antibiotic resistome and microbial community under different TCS concentrations in activated sludge system were evaluated in this study.The experiment was conducted in a sequencing batch reactor(SBR)for 240 days.Quantitative PCR results demonstrated that the abundance of ammonium oxidizing bacteria could be temporarily inhibited by 1 mg/L TCS and then gradually recovered.And the abundances of nitrite oxidizing bacteria(NOB) under 2.5 and 4 mg/L TCS were three orders of magnitude lower than that of seed sludge,which accounted for partial nitrification.When the addition of TCS was stopped,the abundance of NOB increased.The mass balance experiments of TCS demonstrated that the primary removal pathway of TCS changed from adsorption to biodegradation as TCS was continuously added into the SBR system.Moreover,TCS increased the abundance of mexB,indicating the efflux pump might be the main TCS-resistance mechanism.As a response to TCS,bacteria could secrete more protein(PN) than polysaccharide.Three-dimensional excitation-emission matrix revealed that tryptophan PN-like substances might be the main component in PN to resist TCS.High-throughput sequencing found that the relative abundances of Paracoccus,Pseudoxanthomonas and Thauera increased,which could secrete extracellular polymeric substances(EPS).And Sphingopyxis might be the main TCS-degrading bacteria.Overall,TCS could cause partial nitrification and increase the relative abundances of EPS-secreting bacteria and TCS-degrading bacteria.     

Cornmeal-induced resistance to ciprofloxacin and erythromycin in enterococci

Triclosan is used as an antibacterial agent in household items and personal care products. Since this compound is found in maternal milk of humans and bodies of wild animals, there is growing concern among some consumer groups and scientific community that triclosan is adverse for humans and wild animals. In order to estimate adverse actions of triclosan, the effects of triclosan on intracellular Zn²⁺ concentration and cellular thiol content were studied in rat thymocytes by the use of flow cytometer with appropriate fluorescent probes. Triclosan at 1-3 μM (sublethal concentrations) increased the intensity of FluoZin-3 fluorescence (intracellular Zn²⁺ concentration) and decreased the intensity of 5-chloromethylfluorescein (5-CMF) fluorescence (cellular thiol content). Negative correlation (r = −0.985) between triclosan-induced changes in FluoZin-3 and 5-CMF fluorescences was found. Removal of external Zn²⁺ did not significantly affect the triclosan-induced augmentation of FluoZin-3 fluorescence, suggesting an intracellular Zn²⁺ release by triclosan. These actions of triclosan were similar to those of H₂O₂ and triclosan significantly potentiated the cytotoxicity of H₂O₂. Therefore, the results may suggest that triclosan at sublethal concentrations induces oxidative stress that decreases cellular thiol content, resulting in an increase in intracellular Zn²⁺ concentration by Zn²⁺ release from intracellular store(s). Since recent studies show many physiological roles of intracellular Zn²⁺ in cellular functions, the triclosan-induced disturbance of cellular Zn²⁺ homeostasis may induce adverse actions on the cells.     

Investigation on the interaction between triclosan and bovine serum albumin by spectroscopic methods

Abstract

Multi-spectroscopic and molecular docking methods were used to study the interaction between triclosan (TCS) and bovine serum albumin (BSA). The results indicated that the fluorescence quenching of BSA by TCS was due to the formation of TCS–BSA complex through static quenching. This result was also demonstrated by time-resolved fluorescence experiment. The binding constants and number of binding sites between TCS and BSA were 1.30?×?10⁵ M^?1 and 1.17 at 298?K, respectively. The thermodynamic parameters were studied in detail which suggested that hydrophobic forces and hydrogen bond played major roles in the TCS–BSA interaction. Moreover, the site marker competitive experiments and docking studies revealed that TCS could bind BSA into site I in subdomain IIA. All the results of UV–vis spectrophotometry, circular dichroism spectroscopy and synchronous fluorescence spectroscopy showed that interaction between TCS and BSA induced conformation changes of BSA.     

Comparison of two screening level risk assessment approaches for six disinfectants and pharmaceuticals

For three examples of both groups (the disinfectants biphenylol, 4-chloro-m-cresol and triclosan and the pharmaceuticals ivermectin, ibuprofen and oxytetracycline) a relative initial risk assessment (RIRA) was performed assuming a standard emission of 1 kg/d to the most relevant environmental compartment. In addition the hazard of the compounds was evaluated based upon their persistence, toxicity and bioaccumulative properties (PTB). Both estimated and measured parameters were used for this purpose. In addition to an analysis of the risks of the pharmaceuticals and disinfectants per se, the capacity to discern between the intrinsic risk of different compounds is evaluated for both criteria used. It is concluded that the RIRA has a higher discriminative value and yields more information compared to the PTB-criterion.     

Biological degradation of triclocarban and triclosan in a soil under aerobic and anaerobic conditions and comparison with environmental fate modelling

Triclocarban and triclosan are two antimicrobial agents widely used in many personal care products. Their biodegradation behaviour in soil was investigated by laboratory degradation experiments and environmental fate modelling. Quantitative structure-activity relationship (QSAR) analyses showed that triclocarban and triclosan had a tendency to partition into soil or sediment in the environment. Fate modelling suggests that either triclocarban or triclosan "does not degrade fast" with its primary biodegradation half-life of "weeks" and ultimate biodegradation half-life of "months". Laboratory experiments showed that triclocarban and triclosan were degraded in the aerobic soil with half-life of 108 days and 18 days, respectively. No negative effect of these two antimicrobial agents on soil microbial activity was observed in the aerobic soil samples during the experiments. But these two compounds persisted in the anaerobic soil within 70 days of the experimental period.     

Distribution of bisphenol-A, triclosan and n-nonylphenol in human adipose tissue, liver and brain

In this study, an analytical method was optimized for the determination of bisphenol-A (BPA), triclosan (TCS) and 4-n-nonylphenol (4n-NP), environmental contaminants with potential endocrine disruptive activities, in human tissues. The method consisted of a liquid extraction step, derivatization with pentafluorobenzoylchloride followed by a clean-up on acidified silica and detection with gas chromatography coupled with mass spectrometry (GC-ECNI/MS). Recoveries ranged between 92% and 102% with a precision below 5%. Limits of quantification ranged between 0.3-0.4 ng g⁻¹, 0.045-0.06 ng g⁻¹ and 0.003-0.004 ng g⁻¹ for BPA, TCS and 4n-NP in different tissues, respectively. The method was applied for the determination of BPA, TCS and 4n-NP in paired adipose tissue, liver and brain samples from 11 individuals. BPA could be detected in almost all tissues, with the highest concentrations found in adipose tissue (mean 3.78 ng g⁻¹), followed by liver (1.48 ng g⁻¹) and brain (0.91 ng g⁻¹). TCS showed the highest concentrations in liver (3.14 ng g⁻¹), followed by adipose tissue (0.61 ng g⁻¹), while it could be detected in only one brain sample. Levels of 4n-NP were much lower, mostly undetected, and therefore 4n-NP is considered of minor importance for human exposure. Despite the measurable concentrations in adipose tissue, these compounds seem to have a low bioaccumulation potential. The reported concentrations of free BPA in the various tissues are slight disagreement with pharmacokinetic models in humans and rats and therefore the possibility of external contamination with BPA during sample collection/storage cannot be ruled out.     

Photodegradation of the synthetic fragrance OTNE and the bactericide triclosan adsorbed on dried loamy sand--results from models and experiments

Fragrances such as OTNE (marketed as Iso-E-Super®) and bactericides such as triclosan (marketed as Igrasan) are present in waste water and thus finally sorbed to sewage sludge. With that sludge they can reach agricultural fields where they potentially can undergo photodegradation processes. In this study the photodegradation of OTNE and triclosan on dried loamy sand was measured under artificial sunlight conditions in laboratory experiments. These compounds were artificially added with concentrations of 1 μg g⁻¹ on pre-rinsed dried loamy sand. The decrease in concentration with light irradiation was measured for 32 d in comparison to soil samples without light irradiation. The estimated light source intensity was 27 W m⁻². Within the experiment, the apparent half-life was 7 and 17 d for OTNE and triclosan respectively. The decrease did not simply follow first-order kinetics. The apparent rate constant decreased in the latter stage of reaction, suggesting that part of the chemicals were inaccessible for degradation. Two models, i.e., a diffusion-limited model, and a light penetration-limited model, were used in comparison to the measured data to explain the observed degradation limitations in the latter stages of the experiments. Comparing the hereby obtained model parameters with estimated physico-chemical parameters for the soil and the two chemical compounds, the light penetration-limited model, in which the degradation in the soil surface layer is assumed to be limited due to the shading effect of light in the upper thin soil layer, showed to be the most realistic in describing the photodegradation.     

Removal and formation of chlorinated triclosan derivatives in wastewater treatment plants using chlorine and UV disinfection

Triclosan, a common antimicrobial agent, may react during the disinfection of wastewater with free chlorine to form three chlorinated triclosan derivatives (CTDs). This is of concern because the CTDs may be photochemically transformed to tri- and tetra-chlorinated dibenzo-p-dioxins when discharged into natural waters. In this study, wastewater influent, secondary (pre-disinfection) effluent, and final (post-disinfection) effluent samples were collected on two occasions each from two activated sludge wastewater treatment plants, one using chlorine disinfection and one using UV disinfection. Concentrations of triclosan and three CTDs were determined using ultra performance liquid chromatography-triple quadrupole mass spectrometry with isotope dilution methodology. Triclosan and the CTDs were detected in every influent sample at levels ranging from 453 to 4530 and 2 to 98 ng L⁻¹, respectively, though both were efficiently removed from the liquid phase during activated sludge treatment. Triclosan concentrations in the pre-disinfection effluent ranged from 36 to 212 ng L⁻¹, while CTD concentrations were below the limit of quantification (1 ng L⁻¹) for most samples. In the treatment plant that used chlorine disinfection, triclosan concentrations decreased while CTDs were formed during chlorination, as evidenced by CTD levels as high as 22 ng L⁻¹ in the final effluent. No CTDs were detected in the final effluent of the treatment plant that used UV disinfection. The total CTD concentration in the final effluent of the chlorinating treatment plant reached nearly one third of the triclosan concentration, demonstrating that the chlorine disinfection step played a substantial role in the fate of triclosan in this system.     

Effects of growth substrate on triclosan biodegradation potential of oxygenase-expressing bacteria

Triclosan is an antimicrobial agent, an endocrine disrupting compound, and an emerging contaminant in the environment. This is the first study investigating triclosan biodegradation potential of four oxygenase-expressing bacteria: Rhodococcus jostii RHA1, Mycobacterium vaccae JOB5, Rhodococcus ruber ENV425, and Burkholderia xenovorans LB400. B. xenovorans LB400 and R. ruber ENV425 were unable to degrade triclosan. Propane-grown M. vaccae JOB5 can completely degrade triclosan (5 mg L⁻¹). R. jostii RHA1 grown on biphenyl, propane, and LB medium with dicyclopropylketone (DCPK), an alkane monooxygenase inducer, was able to degrade the added triclosan (5 mg L⁻¹) to different extents. Incomplete degradation of triclosan by RHA1 is probably due to triclosan product toxicity. The highest triclosan transformation capacity (T_c, defined as the amount of triclosan degraded/the number of cells inactivated; 5.63 × 10⁻³ ng triclosan/16S rRNA gene copies) was observed for biphenyl-grown RHA1 and the lowest T_c (0.20 × 10⁻³ ng-triclosan/16S rRNA gene copies) was observed for propane-grown RHA1. No triclosan degradation metabolites were detected during triclosan degradation by propane- and LB + DCPK-grown RHA1. When using biphenyl-grown RHA1 for degradation, four chlorinated metabolites (2,4-dichlorophenol, monohydroxy-triclosan, dihydroxy-triclosan, and 2-chlorohydroquinone (a new triclosan metabolite)) were detected. Based on the detected metabolites, a meta-cleavage pathway was proposed for triclosan degradation.     

Occurrence and distribution of triclosan in the German Bight (North Sea)

The potential of triclosan (TCS) acting as an endocrine disruptor has led to growing concern about the presence of TCS in the environment. In this study, seawater samples were collected from the German Bight during sampling campaigns conducted with the German research ships Gauss and Ludwig Prandtl. TCS was determined both in the dissolved phase and in the suspended particulate matters with concentrations ranging 0.8-6870 pg L⁻¹ and <1-95 pg L⁻¹, respectively. High concentrations of TCS were present in the estuaries of the Elbe and the Weser, indicating significant input of TCS by the river discharge. The correlation coefficient (R²) between the dissolved concentration and salinity was 0.79 for the data obtained from the Gauss cruise, showing an obvious declining trend from the coast to the open sea.