Tuning the Locally Enhanced Electric Field Treatment (LEEFT) between Electrophysical and Electrochemical Mechanisms for Bacteria Inactivation.

Efficient drinking water disinfection methods are critical for public health. Locally enhanced electric field treatment (LEEFT) is an antimicrobial method that uses sharp structures, like metallic nanowires, to enhance the electric field at tips and cause bacteria inactivation. Electroporation is the originally designed mechanism of LEEFT. Although oxidation is typically undesired due to byproduct generation and electrode corrosion, it can enhance the overall disinfection efficiency. In this work, we conduct an operando investigation of LEEFT, in which we change the electrical parameters to tune the mechanisms between electrophysical electroporation and electrochemical oxidation. Pure electroporation (i.e., without detectable oxidation) could be achieved under a duty cycle of ≤0.1% and a pulse width of ≤2 µs. Applying 2 µs pulses at 7-8 kV/cm and 0.1% duty cycle results in 80-100% bacteria inactivation with pure electroporation. A higher chance of oxidation is found with a higher duty cycle and a longer pulse width, where the antimicrobial efficiency could also be enhanced. For water with a higher conductivity, a higher antimicrobial efficiency can be achieved under the same treatment conditions, and electrochemical reactions could be induced more easily. The findings shown in this work improve the fundamental understanding of LEEFT and help optimize the performance of LEEFT in real applications.

Synthesis of Luminescent Copper Nanoparticles Using Couroupita guianensis Flower Extract: Evaluation of Antibacterial and Anticancer Activities.

The biosynthesis of nanoparticles is a crucial research area aimed at developing innovative, cost-effective, and eco-friendly synthesis techniques for various applications. Herein, we synthesized copper oxide nanoparticles (CuNPs) using Couroupita guianensis flower extract via a simple green synthesis method. These green CuNPs demonstrate promising antimicrobial activity and anticancer activity against A549 nonsmall cell lung cancer (NSCLC) cells. We comprehensively characterized the CuNPs using UV spectrum, XRD, FTIR, SEM, and EDS analyses. The antibacterial and anticancerous performance is attributed to their spherical-like morphology, which enhances effective interaction with bacterial and cancer cells. Moreover, CuNPs proved effective in inactivating Escherichia coli, achieving 2%, 52%, and 99% inactivation at 0, 30, and 60 min, respectively, and Listeria monocytogenes, achieving 1%, 48%, and 98% inactivation at 0, 30, and 60 min, respectively, under visible light. Furthermore, the CuNPs exhibited significant anticancer activity against A549 NSCLC cells, achieving cell viability reductions of 10%, 30%, 50%, 70%, 83%, and 91% at concentrations of 25, 50, 100, 150, 200, and 250 µg/mL, respectively. The green synthesized CuNPs demonstrate their potential in biomedical applications.

Self-Powered Disinfection Using Triboelectric, Conductive Wires of Metal-Organic Frameworks.

Efficient water disinfection is vitally needed in rural and disaster-stricken areas lacking power supplies. However, conventional water disinfection methods strongly rely on external chemical input and reliable electricity. Herein, we present a self-powered water disinfection system using synergistic hydrogen peroxide (H2O2) assisted electroporation mechanisms driven by triboelectric nanogenerators (TENGs) that harvest electricity from the flow of water. The flow-driven TENG, assisted by power management systems, generates a controlled output with aimed voltages to drive a conductive metal-organic framework nanowire array for effective H2O2 generation and electroporation. The injured bacteria caused by electroporation can be further damaged by facile diffused H2O2 molecules at high throughput. A self-powered disinfection prototype enables complete disinfection (>99.9999% removal) over a wide range of flows up to 3.0 × 104 L/(m2 h) with low water flow thresholds (200 mL/min; ∼20 rpm). This rapid, self-powered water disinfection method is promising for pathogen control.

Standardized Figures of Merit for Proper Benchmarking of Photocatalytic Inactivation of Bacteria Using Thin Films Based on TiO2 Nanostructures.

The study of photocatalysts fixed to surfaces for the inactivation of bacteria in wastewater has increased in recent years. However, there are no standardized methods to analyze the photocatalytic antibacterial activity of these materials, and no systematic studies have attempted to relate this activity to the number of reactive oxygen species generated during UV-light irradiation. Additionally, studies regarding photocatalytic antibacterial activity are usually carried out with varying pathogen concentrations, UV light doses, and catalyst amounts, making it difficult to compare results across different materials. The work introduces the photocatalytic bacteria inactivation efficiency (PBIE) and bacteria inactivation potential of hydroxyl radicals (BIPHR) figures of merit for evaluating the photocatalytic activity of catalysts fixed onto surfaces for bacteria inactivation. To demonstrate their applicability, these parameters are calculated for various photocatalytic TiO2 -based coatings, accounting for the catalyst area, the kinetic reaction rate constant associated with bacteria inactivation and hydroxyl radical formation, reactor volume, and UV light dose. This approach enables a comprehensive comparison of photocatalytic films prepared by different fabrication techniques and evaluated under diverse experimental conditions, with potential applications in the design of fixed-bed reactors.

Photocatalytic investigation of textile dyes and E. coli bacteria from wastewater using Fe3O4@MnO2 heterojunction and investigation for hydrogen generation on NaBH4 hydrolysis.

Various impurities found nowadays in water can be detrimental to human health. This work focused on utilizing Fe3O4@MnO2 nanocomposite for cleaning organic contaminants from water, including rhodamine B (RhB) and Escherichia coli (E. coli). Analysis methods such as XRD, UV-vis, TEM, and FTIR were used to describe the nanocomposite. The results showed that the developed nanocomposite has good photocatalytic activity against pollutants in wastewater. The E. coli was destroyed after 90 min, and the RhB photodegradation rate was 75%. Moreover, the Fe3O4@MnO2 efficiency as a catalyst for producing hydrogen as an alternative energy source was tested. According to the calculations, the nanomaterial's turnover frequency, activation energy, enthalpy, and entropy are 1061.3 h-1, 28.93 kJ/mol, 26.38 kJ/mol, and -128.41 J/mol.K, respectively. Four reusability tests were completed, and the average reusability was 78%. The obtained data indicated the excellent potential for the developed Fe3O4@MnO2 nanomaterial to act as an adsorbent, thus representing an alternative to the classical depollution methods. This study showed that nanoparticles have a photocatalytic effect against pathogenic bacteria and RhB azo dye in polluted waters and offer an effective catalytic activity to produce hydrogen as an alternative energy source.

A tryptophan synchronous and normal fluorescence study on bacteria inactivation mechanism.

The UV photodissociation kinetics of tryptophan amino acid, Trp, attached to the membrane of bacteria, Escherichia coli and Bacillus subtilis, have been studied by means of normal and synchronous fluorescence. Our experimental data suggest that the fluorescence intensity of Trp increases during the first minute of irradiation with 250 nm to ∼ 280 nm, 7 mW/cm2 UV light, and subsequently decreases with continuous irradiation. During this short, less than a minute, period of time, 70% of the 107 cell per milliliter bacteria are inactivated. This increase in fluorescence intensity is not observed when tryptophan is in the free state, namely, not attached to a protein, but dissolved in water or saline solution. This increase in fluorescence is attributed to the additional fluorescence of tryptophan molecules formed by protein unfolding, the breakage of the bond that attaches Trp to the bacterial protein membrane, or possibly caused by the irradiation of 2 types of tryptophan residues that photolyze with different quantum yields.

Advances in 2D MXenes-based materials for water purification and disinfection: Synthesis approaches and photocatalytic mechanistic pathways.

MXenes two-dimensional materials have recently excited researchers' curiosity for various industrial applications. MXenes are promising materials for environmental remediation technologies to sense and mitigate various intractable hazardous pollutants from the atmosphere due to their inherent mechanical and physicochemical properties, such as high surface area, increased hydrophilicity, high conductivity, changing band gaps, and robust electrochemistry. This review discusses the versatile applications of MXenes and MXene-based nanocomposites in various environmental remediation processes. A brief description of synthetic procedures of MXenes nanocomposites and their different properties are highlighted. Afterward, the photocatalytic abilities of MXene-based nanocomposites for degrading organic pollutants, removal of heavy metals, and inactivation of microorganisms are discussed. In addition, the role of MXenes anti-corrosion support in the lifetime of some semiconductors was addressed. Current challenges and future perspectives toward the application of MXene materials for environmental remediation and energy production are summarized for plausible real-world use.

In situ detection of live-to-dead bacteria ratio after inactivation by means of synchronous fluorescence and PCA.

The determination of live and dead bacteria is of considerable significance for preventing health care-associated infection in hospitals, field clinics, and other areas. In this study, the viable (live) and nonviable (dead) bacteria in a sample were determined by means of their fluorescence spectra and principal component analysis (PCA). Data obtained in this study show that it is possible to identify bacteria strains and determine the live/dead ratio after UV light inactivation and antibiotic treatment, in situ, within minutes. In addition, synchronous fluorescence scans enable the identification of bacterial components such as tryptophan, tyrosine, and DNA. Compared with the time-consuming plating and culturing methods, this study renders a means for rapid detection and determination of live and dead bacteria.

Investigating the Use of Impedance Flow Cytometry for Classifying the Viability State of E. coli.

Bacteria detection, counting and analysis is of great importance in several fields. When viability plays a major role in decision making, the counting of colony-forming units grown on agar plates remains the gold standard. However, because plate counts depend on the growth of the bacteria, it is a slow procedure and only works with culturable species. Impedance flow cytometry (IFC) is a promising technology for particle detection, counting and characterization. It relies on the perturbation of an electric field by particles flowing through a microfluidic channel. The perturbation is directly related to the electrical properties of the particles, and therefore provides information about their composition and structure. In this work we investigate whether IFC can be used to differentiate viable cells from inactivated cells. Our findings demonstrate that the specific viability state of the bacteria has to be considered, but that with proper characterization thresholds, IFC can be used to classify bacterial viability states. By using three different inactivation methods-ethanol, heat and autoclavation-we have been able to show that the impedance response of Escherichia coli depends on its viability state, but that the specific response depends on the inactivation method. With these findings we expect to be able to optimize IFC for more reliable bacteria detection and counting in the future.

Cold plasma inactivation of chronic wound bacteria.

Cold plasma is partly ionized non-thermal plasma generated at atmospheric pressure. It has been recognized as an alternative approach in medicine for sterilization of wounds, promotion of wound healing, topical treatment of skin diseases with microbial involvement and treatment of cancer. Cold plasma used in wound therapy inhibits microbes in chronic wound due to its antiseptic effects, while promoting healing by stimulation of cell proliferation and migration of wound relating skin cells. In this study, two types of plasma systems are employed to generate cold plasma: a parallel plate dielectric barrier discharge and a capillary-guided corona discharge. Parameters such as applied voltage, discharge frequency, treatment time and the flow of the carrier gas influence the cold plasma chemistry and therefore change the composition and concentration of plasma species that react with the target sample. Chronic wound that fails to heal often infected by multidrug resistant organisms makes them recalcitrant to healing. Methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa (Pseudomonas aeruginosa) are two common bacteria in infected and clinically non-infected wounds. The efficacies of the cold plasma generated by the two designs on the inactivation of three different isolates of MRSA and four isolates of P. aeruginosa are reported here.

Water disinfection using hydrogen peroxide with fixed bed hematite catalyst - kinetic and activity studies.

A lab-scale reactor with a fixed-bed hematite catalyst for the effective decomposition of H2O2 and bacteria inactivation was designed. The bactericidal effect is the largest at a low initial bacterial count of 2·103 CFU/L, which is typical for natural surface waters. When using a 5 mM H2O2 solution and a residence time of 104 min, the reduction in the number of E. coli bacteria is about 3.5-log. At a higher initial bacterial count of 1-2·104 CFU/L, a 5 mM H2O2 solution reduces the bacteria number by about 4-log. The H2O2 decomposition follows the log-linear kinetics of a first-order reaction while the bacterial inactivation does not. The kinetics of bacterial inactivation was described using the Weibull model in the modified form: log10(N0/N) = b · tn. The values of the non-linearity parameter n were found to be lower than 1, indicating that bacterial inactivation slows down over time. With increasing initial H2O2 concentration, the rate parameter b increases while the non-linearity parameter n decreases. With increasing temperature, both parameters increase. The stability of the catalyst has been proved by XRD, FTIR, SEM, and ICP-OES. The concentration of iron leaching into water during disinfection is much lower than the limit declared by WHO for iron in drinking water. The results show that technical-grade hematite is a promising Fenton-like catalyst for water disinfection. The fixed-bed reactor can be the basis of the mobile installations for water purification in emergencies.

Dielectric barrier discharge plasma promotes disinfection-residual-bacteria inactivation via electric field and reactive species.

Traditional disinfection processes face significant challenges such as health and ecological risks associated with disinfection-residual-bacteria due to their single mechanism of action. Development of new disinfection processes with composite mechanisms is therefore urgently needed. In this study, we employed liquid ground-electrode dielectric barrier discharge (lgDBD) to achieve synergistic sterilization through electric field electroporation and reactive species oxidation. At a voltage of 12 kV, Pseudomonas fluorescens (ultraviolet and ozone-resistant) and Bacillus subtilis (chlorine-resistant) were completely inactivated within 8 and 6 min, respectively, surpassing a 7.0-log reduction. The lgDBD process showed good disinfection performance across a wide range of pH values and different practical water samples. Staining experiments suggest that cellular membrane damage contributes to this inactivation. In addition, we used a two-dimensional parallel streamer solver with kinetics code to fashion a representative model of the basic discharge unit, and discovered the presence of a persistent electric field during the discharge process with a peak value of 2.86 × 106 V/m. Plasma discharge generates excited state species such as O(1D) and N2(C3Πu), and further forms reactive oxygen and nitrogen species at the gas-liquid interface. The physical process, which is driven by electric field-induced cell membrane electroporation, synergizes with the bactericidal effects of reactive oxygen and nitrogen species to provide effective disinfection. Adopting the lgDBD process enhances sterilization efficiency and adaptability, underscoring its potential to revolutionize physicochemical synergistic disinfection practices.

In-situ Hf/Zr co-doped Fe2O3 nanorod decorated with CuOx/CoOx: Enhanced photocatalytic performance for antibacterial and organic pollutants.

Herein, we successfully synthesized Hf/Zr co-doping on Fe2O3 nanorod photocatalyst by a hydrothermal process and quenching methods. The synergistic roles of Hf and Zr double-doping on the bacteria inactivation test and decomposition of organic pollutants were investigated in detail for the 1 wt% CoOx loaded Hf/Zr-Fe2O3 NRs and CuOx/CoOx loaded Hf/Zr-Fe2O3 NRs photocatalyst. Initially, the rod-like porous morphology of the Hf/Zr-doped Fe2O3 NRs was produced via a hydrothermal method at various Hf co-doping (0, 2, 4, 7 and 10)%. Further, CoOx and CuOx loaded by a wet impregnation approach on the Hf/Zr-Fe2O3 NRs and a highly photoactive Hf(4)/Zr-Fe2O3 [CoOx/CuOx] NRs photocatalyst were developed. After the Hf(4)/Zr-Fe2O3 [CoOx/CuOx] NRs photocatalyst treatment, the Bio-TEM imagery of bacterial cells showed extensive morphological deviations in cell membranes. Hf(4)/Zr-Fe2O3 NR achieved 84.1% orange II degradation upon 3 h illumination, which is higher than that of Hf-Fe2O3 and Zr-Fe2O3 (68.7 and 73.5%, respectively). Additionally, the optimum sample, Hf(4)/Zr-Fe2O3 [CoOx/CuOx] photocatalyst, exhibited 95.5% orange II dye degradation after light radiation for 3 h. Optimized Hf(4)/Zr-Fe2O3 [CoOx/CuOx] catalysts exhibited 99.9% and 99.7% inactivation of E. coli and S. aureus with 120 min, respectively. Further, scavenger experiments revealed that the electrons are the primary responsible species for photocatalytic kinetics. This work will provide a rapid method for the development of high photocatalytic performance materials for bacterial disinfection and organic degradation.

Phytochemicals-based edible coating for photodynamic preservation of fresh-cut apples.

Nature-derived chemicals have recently gained increased attention to settle down the challenges in the food industry. Quercetin has long been used as a natural medicine but its photoactivity has been neglected. In this work, by combining photodynamic bacteria inactivation (PDI) with an edible coating (Pectin/Quercetin) derived from FDA-approved chemicals, extend shelf-life and protected commercial quality of fresh-cut apples were achieved. Firstly, the potential photoactivated antibacterial performance of Quercetin (a natural plant flavonoid) was clarified with the treatment of a simulated sunlight lamp, realizing antibacterial efficacy of 100 % towards S. aureus (50 min) and L. monocytogenes (80 min) with light treatment. To develop safe and effective preservation of fresh-cut apples, Pectin/Quercetin edible coatings with 100 µmol/L quercetin were adopted. The results showed that the prepared edible coatings form a protective barrier over the surface of apples, effectively resisting bacterial infection and extending shelf life to 10 days while maintaining good commercial quality (including preferable color, keeping 100 % hardness, 80 % sugar content and 17.3 % weightlessness rate). Therefore, the prepared light-driven Pectin/Quercetin in this work has the potential to develop as fresh-cut fruit preservation technology.

Innovative Antibacterial, Photocatalytic, Titanium Dioxide Microstructured Surfaces Based on Bacterial Adhesion Enhancement.

Bacterial colonization and biofilm formation are found on nearly all wet surfaces, representing a serious problem for both human healthcare and industrial applications, where traditional treatments may not be effective. Herein, we describe a synergistic approach for improving the performance of antibacterial surfaces based on microstructured surfaces that embed titanium dioxide nanoparticles (TiO2 NPs). The surfaces were designed to enhance bacteria entrapment, facilitating their subsequent eradication by a combination of UVC disinfection and TiO2 NPs photocatalysis. The efficacy of the engineered TiO2-modified microtopographic surfaces was evaluated using three different designs, and it was found that S2-lozenge and S3-square patterns had a higher concentration of trapped bacteria, with increases of 70 and 76%, respectively, compared to flat surfaces. Importantly, these surfaces showed a significant reduction (99%) of viable bacteria after just 30 min of irradiation with UVC 254 nm light at low intensity, being sixfold more effective than flat surfaces. Overall, our results showed that the synergistic effect of combining microstructured capturing surfaces with the chemical functionality of TiO2 NPs paves the way for developing innovative and efficient antibacterial surfaces with numerous potential applications in the healthcare and biotechnology market.

Visible light-driven photocatalytic bacterial inactivation on PPE, supported by the DFT and bactericidal study.

A novel ZnO-MoO3-ZnMoO3@graphene GZM composite catalyst prepared by microwave hydrothermal process for personal protective equipment textiles (PPE) is presented in this study. The results indicated that the GZM with defect vacancy sites of two types as observed by EPR showed significantly superior inactivation of the E. coli bacteria compared to GZM without the lower defect vacancy sites and concomitant lower electron densities. Photocatalytic activated oxidation by the GZM composites coatings was observed to proceed in acceptable times as well as the bacterial inactivation (log bact. C/Co > 107 within 3 h). Defect sites in the GZM seem to be important leading to the bacterial inactivation process. DFT calculations on the GZM with and without catalyst defect sites were carried out. The electron densities were estimated by the Fourier mapping. The results found in this study showed the potential of GZM-PPE for practical applications.

Synergistic role of in-situ Zr-doping and cobalt oxide cocatalysts on photocatalytic bacterial inactivation and organic pollutants removal over template-free Fe2O3 nanorods.

Herein, we synthesized in-situ Zr-doped Fe2O3 NRs photocatalyst by successive simple hydrothermal and air quenching methods. The synergistic roles of CoOx (1 wt%) and Zr-doping on bacteria inactivation and model organic pollutants over Fe2O3 NRs photocatalyst were studied in detail. Initially, rod-like Zr ((0-8) %)-doped Fe2O3 NRs were produced via a hydrothermal method. CoOx was loaded onto the Zr ((0-8) %)-doped Fe2O3 NRs) surface by a wet impregnation approach. The Zr-doping conditions and CoOx loadings were judiciously optimized, and a highly photoactive CoOx(1 wt%)/Zr(6%)-doped Fe2O3 NRs photocatalyst was developed. The CoOx(1 wt%) loaded Zr(6%)-doped Fe2O3 NRs photocatalyst revealed 99.4% inactivation efficiency compared with (0, 4 and 8)% Zr-doped Fe2O3 NRs, respectively. After CoOx(1 wt%)/Zr(6%)-doped Fe2O3 NRs photocatalyst treatment, Bio-TEM images of bacterial cells showed extensive morphological deviations in cell membranes, compared with the non-treated ones. Additionally, the optimum CoOx(1 wt%)/Zr(6%)-doped Fe2O3 NRs photocatalyst exhibited 99.2% BPA and 98.3% orange II dye degradation after light radiation for 3 h. This work will provide a rapid method for the development of photostable catalyst materials for bacterial disinfection and organic degradation.

Effects of High-Pressure Processing, UV-C Irradiation and Thermoultrasonication on Donor Human Milk Safety and Quality.

Holder pasteurization (HoP) is the current recommended treatment for donor human milk. Although this method inactivates microbial contaminants, it also negatively affects various milk components. High-pressure processing (HPP, 400, 500, and 600 MPa), ultraviolet-C irradiation (UV-C, 2,430, 3,645, and 4,863 J/L) and thermoultrasonication (TUS, 1,080 and 1,620 kJ/L) were investigated as alternatives to thermal pasteurization (HoP). We assessed the effects of these methods on microbiological safety, and on concentration and functionality of immunoglobulin A, lactoferrin, lysozyme and bile salt-stimulated lipase, with LC-MS/MS-based proteomics and activity assays. HoP, HPP, TUS, and UV-C at 4863 J/L, achieved >5-log10 microbial reduction. Native protein levels and functionality showed the highest reduction following HoP, while no significant reduction was found after less intense HPP and all UV-C treatments. Immunoglobulin A, lactoferrin, and lysozyme contents were also preserved after low intensity TUS, but bile salt-stimulated lipase activity was significantly reduced. This study demonstrated that HPP and UV-C may be considered as suitable alternatives to HoP, since they were able to ensure sufficient microbial inactivation while at the same time better preserving the bioactive components of donor human milk. In summary, our results provide valuable insights regarding the evaluation and selection of suitable processing methods for donor human milk treatment, which may replace HoP in the future.

Solar-driven free chlorine advanced oxidation process for simultaneous removal of microcontaminants and microorganisms in natural water at pilot-scale.

Contamination of natural water (NW) by emerging contaminants has been widely pointed out as one of the main challenges to ensure high-quality drinking water. Thus, the effectiveness of a solar-driven free chlorine advanced oxidation process simultaneously investigating the elimination of six organic microcontaminants (OMCs) and three bacteria from NW at a pilot-scale was evaluated in this study. Firstly, the solar/free chlorine process was studied at lab-scale using a solar simulator to evaluate the effect of free chlorine concentration (0.5-10 mg L-1) on OMC degradation and generation of toxic oxyanions (e.g., ClO3- ions). Thus, the best free chlorine concentration observed was applied for the simultaneous removal of OMCs and pathogens under natural solar light at pilot scale. At lab-scale, the solar/free chlorine (2.5 mg L-1) process achieved 80% of total degradation in 5 min (1.4 kJ L-1 of accumulative UV energy) with an oxidant consumption of 0.3 mg L-1 and without ClO3- generation. Similar results were attained under natural solar irradiation at a pilot-scale. For all bacteria strains, the legally required detection limit (DL = 1 CFU 100 mL-1) for reclaimed water reuse was attained in a short contact time. Still, more importantly, the solar/free chlorine (2.5 mg L-1) process effectively avoided the possible bacterial regrowth in the post-treated sample after six days. Finally, the combination of free chlorine with solar irradiation provided a simple and energy-efficient process for OMC and bacteria removal in NW at a pilot-scale.

Photoelectrocatalytic inactivation mechanism of E. coli DH5α (TET) and synergistic degradation of corresponding antibiotics in water.

The occurrence and proliferation of antibiotic-resistance genes (ARGs) / antibiotic-resistant bacteria (ARB) have been currently aggravating due to the increase of antibiotic residues in the aquatic environment. The interaction of ARB/ARGs with antibiotics inevitably occurred during water purification, yet their synergistic purification mechanism remains unclear. Herein, a systematic approach was developed to understand, in-depth, the synergistic mechanism in the coexisted E. coli DH5α (TET) inactivation and tetracycline hydrochloride (TET) degradation using photoelectrocatalysis (PEC) as a model technology. Results showed that low dosage (0 - 40 ppm) of TET exerted a negative influence on ARB inactivation with prolonged bactericidal time from 60 to 160 min. Addition of TET in environmental concentration (5 - 60 ppm) resulted in sub-lethal damage and prolonged PEC treatment time (100 - 160 min), accounting for inhibition effects on ARB inactivation. The major reactive species (RSs) involved in ARB inactivation and TET degradation were evidenced as photogenerated hole, •OH and O2•-, whereas hole and O2•- were demonstrated to be the major disinfectants for ARB/ARG inactivation. The bacterial defense system displayed increased antioxidative activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) to protect ARB cells against oxidative stress. Exposure to 60 ppm TET was a threshold where certain ARB cells were induced into viable but nonculturable bacterial cell (VBNC) state, as evidenced by plate counting and ATP activity analysis, together with the integral cell membranes observed by flow cytometry (FCM) and scanning electron microscope (SEM). These findings appeal for appropriate technical adjustments for water and wastewater treatment to ensure safety of water.