Inactivation of Escherichia coli by sonoelectrocatalytic disinfection using TiO2 as electrode

This is the first study to demonstrate sonoelectrocatalytic disinfection using titanium dioxide (TiO₂) as an anode for effective inactivation of Escherichia coli. In brief, a non-woven TiO₂ fabric used as an anode and a platinum cathode were immersed in an E. coli suspension in which a positive potential was applied to TiO₂ concomitant with ultrasound (US) irradiation. Two control experiments were performed using E. coli suspensions to exhibit the effects of the sonoelectrocatalytic disinfection. One was disinfection by applying a positive potential to a TiO₂ electrode, but without US irradiation (electrochemical disinfection). The other was disinfection without applying a potential, but with US irradiation in the presence of TiO₂ (sonocatalytic disinfection). The cell inactivation rate in sonoelectrocatalytic disinfection was synergistically much more enhanced than the combined inactivation rates in electrochemical disinfection and sonocatalytic disinfection. This synergistically enhanced inactivation rate of E. coli cells was attributable to effective reaction of the sonocatalytically generated OH radicals with E. coli cells at the surface of the TiO₂ anode, which resulted from the electroadsorption of E. coli cells toward the TiO₂ anode.     

Electrical sterilization of Escherichia coli by electrostatic atomization with a photo-chemical catalyst

The inactivation of Escherichia coli (E. coli) was studied to find the optimal sterilization conditions using electrostatic atomization with a near ultraviolet light-emitting diode (near UV-LED) and TiO₂ nanofiber films. Three types of near UV-LEDs of different wavelengths (365 nm, 375 nm, 385 nm) were used in this investigation. In order to enhance the LED performances, TiO₂ nanofiber films were utilized for the production of a photo-chemical catalytic effect. In these studies, the flow rate condition of the E. coli mixture was varied from 2 ml/h to 10 ml/h for the purposes of measuring the capabilities of the proposed sterilization method. During these electrostatic atomization experiments, Each LED irradiated a TiO₂ nanofiber film and atomized E. coli mixture. The results of these experiments were compared with electrostatic atomization performed without the use of photo-chemical catalyst effects. The experimental result shows that the optimal sterilization effect of E. coli is 375 nm wavelength with a TiO₂ nanofiber employed during electrostatic atomization.     

Enhanced antimicrobial activity of ZnO nanofluids in sonophotocatalysis and its mechanism

This study investigated the inactivation efficiency of ZnO nanofluids against E. coli in sonophotocatalysis with the aeration of nitrogen, oxygen, argon and their mixtures. The results showed that inactivation efficiency was increased when aeration was combined with sonophotocatalysis. Addition of different types of gases could lead to the different inactivation efficiency. The inactivation efficiencies were shown in the following order: no aeration < nitrogen < argon < oxygen < Ar/O₂(3:7) < Ar/O₂(7:3) < Ar/O₂(5:5). The production of hydroxyl radicals was explored to understand the inactivation mechanism. Compared with sonophotocatalysis without aeration, more hydroxyl radicals were produced in sonophotocatalysis with aeration, which could lead to changes of cellular substances. Furthermore, characterization of E. coli cells using Raman spectroscopy and FTIR illustrated that sonophotocalysis could affect the cellular substances containing carbohydrates, proteins and P containing molecules. Results suggested that the enhanced antimicrobial activity with aeration was originated from stronger cavitational activity, together with the formation of hydroxyl radicals. Compared to sonophotocatalysis without aeration, more dissolved oxygen was existed in sonophotocatalysis with aeration, which could enhance the formation of hydroxyl radicals.     

The effect of ultrasound treatment in combination with nisin on the inactivation of Listeria innocua and Escherichia coli

Ultrasound, alone or in combination with natural antimicrobials, is a novel food processing technology of interest to replace traditional food decontamination methods, as it is milder than classical sterilisation (heat treatment) and maintains desirable sensory characteristics. However, ultrasound efficacy can be affected by food structure/composition, as well as the order in which combined treatments are applied. More specifically, treatments which target different cell components could result in enhanced inactivation if applied in the appropriate order. The microbial properties i.e. Gram positive/Gram negative can also impact the treatment efficacy.This work presents a systematic study of the combined effect of ultrasound and nisin on the inactivation of the bacteria Listeria innocua (Gram positive) and Escherichia coli (Gram negative), at a range of cavitation conditions (44, 500, 1000 kHz). The order of treatment application was varied, and the impact of system structure was also investigated by varying the concentration of Xanthan gum used to create the food model systems (0 – 0.5% w/v). Microbial inactivation kinetics were monitored, and advanced microscopy and flow cytometry techniques were utilised to quantify the impact of treatment on a cellular level.Ultrasound was shown to be effective against E. coli at 500 kHz only, with L. innocua demonstrating resistance to all frequencies studied. Enhanced inactivation of E. coli was observed for the combination of nisin and ultrasound at 500 kHz, but only when nisin was applied before ultrasound treatment. The system structure negatively impacted the inactivation efficacy. The combined effect of ultrasound and nisin on E. coli was attributed to short-lived destabilisation of the outer membrane as a result of sonication, allowing nisin to penetrate the cytoplasmic membrane and facilitate cell inactivation.     

Non-thermal pasteurization of lipid emulsions by combined supercritical carbon dioxide and high-power ultrasound treatment

Supercritical carbon dioxide (SC-CO₂) is a novel method for food pasteurization, but there is still room for improvement in terms of the process shortening and its use in products with high oil content. This study addressed the effect of high power ultrasound (HPU) on the intensification of the SC-CO₂ inactivation of E. coli and B. diminuta in soybean oil-in-water emulsions. Inactivation kinetics were obtained at different pressures (100 and 350 bar), temperatures (35 and 50 °C) and oil contents (0, 10, 20 and 30%) and were satisfactorily described using the Weibull model. The experimental results showed that for SC-CO₂ treatments, the higher the pressure or the temperature, the higher the level of inactivation. Ultrasound greatly intensified the inactivation capacity of SC-CO₂, shortening the process time by approximately 1 order of magnitude (from 50 to 90 min to 5–10 min depending on the microorganism and process conditions). Pressure and temperature also had a significant (p < 0.05) effect on SC-CO₂ + HPU inactivation for both bacteria, although the effect was less intense than in the SC-CO₂ treatments. E. coli was found to be more resistant than B. diminuta in SC-CO₂ treatments, while no differences were found when HPU was applied. HPU decreased the protective effect of oil in the inactivation and similar microbial reductions were obtained regardless of the oil content in the emulsion. Therefore, HPU intensification of SC-CO₂ treatments is a promising alternative to the thermal pasteurization of lipid emulsions with heat sensitive compounds.     

Sonophotocatalytic inactivation of E. coli using ZnO nanofluids and its mechanism

The present study evaluated inactivation efficiency of a sonophotocatalytic process using ZnO nanofluids including ultrasonic parameters such as power density, frequency and time. The result showed that inactivation efficiency was increased by 20% when ultrasonic irradiation was combined with photocatalytic process in the presence of natural light. Comparison of inactivation efficiency in photocatalytic, ultrasonic and sonocatalytic processes using Escherichia coli as a model bacteria identified that inactivation efficiencies are shown in the following order: ultrasonic irradiation < sonocatalysis < photocatalysis < sonophotocatalysis. Furthermore, inactivation mechanism of sonophotocatalysis was proposed. Studies of reactive oxygen species (ROS) and zinc ions (Zn²⁺) release evaluation revealed that ROS play a key role in bacterial inactivation rather than Zn²⁺. Permeability of outer membrane (OM) and inner membrane (IM) of E. coli bacterial cells were studied and exhibited that sonophotocatalysis increased the permeability of OM and IM significantly. The enhanced bacterial inactivation effect in sonophotocatalytic process contributed to acoustic cavitation, sonocatalysis of ZnO and sonoporation phenomenon.     

Effect of ultrasound and chlorine dioxide on Salmonella Typhimurium and Escherichia coli inactivation in poultry chiller tank water

This study evaluated the application of ultrasound alone or combined with chlorine dioxide (ClO₂) for Salmonella Typhimurium and Escherichia coli inactivation in poultry processing chiller tank water. A Full Factorial Design (FFD) 2² was conducted for each microorganism to evaluate the effect of ultrasound exposure time (x₁: 1 to 9 min; fixed: 37 kHz; 330 W; 25 °C) using a bath, and ClO₂ concentration (x₂: 1 to 17 mg L⁻¹) on microorganism count expressed in log CFU mL⁻¹ in distilled water. Variable x₂ had a negative effect on Salmonella Typhimurium (-5.09) and Escherichia coli (-2.00) count, improving the inactivation; while a x₁ increase present no inactivation improvement, explaining the use of x₁ lower level (1 min) and x₂ higher level (17 mg L⁻¹). The best condition for microorganism inactivation based on FFD was evaluated in chiller tank water (with organic matter) at 25, 16, and 4 °C; x₁ was kept (1 min), however x₂ was adjusted to obtain the same residual free chlorine (2.38 mg L⁻¹) considering the ClO₂ consumption by organic matter, achieving the value of 30 mg L⁻¹. An inactivation of 49% and 31% were observed for Salmonella Typhimurium and Escherichia coli. When ultrasound was replaced by a simple agitation in the presence of ClO₂, there was no inactivation for both microorganisms. Moreover, at poultry carcass pre-chilling (16 °C) and chilling (4 °C) conditions, the synergism of ultrasound combined with ClO₂ was more pronounced, with microorganisms’ reductions up to 100%.     

Enhanced antibacterial performance of hybrid semiconductor nanomaterials: ZnO/SnO2 nanocomposite thin films

The nano-sized coupled oxides ZnO/SnO₂ thin films in a molar ratio of 2:1 (Z2S), 1:1 (ZS) and 1:2 (ZS2) were prepared using sol-gel dip coating method and characterized with X-ray diffraction (XRD), scanning electron microscopy (SEM) and UV-vis spectroscopy. Escherichia coli (E. coli, ATCC 25922) was selected as a model for the Gram-negative bacteria to evaluate antibacterial property of composite samples compared with single ZnO (Z) and single SnO₂ (S) films. The antibacterial activity has been studied applying the so-called antibacterial drop test under UV illumination. The bactericidal activity was estimated by relative number of bacteria survived calculated from the number of viable cells which form colonies on the nutrient agar plates. The influence of the SnO₂-ZnO nanocomposite composition on the structural features and on the antibacterial properties of the thin films are reported and discussed. It is found that all coatings exhibited a high antibacterial activity. The coupled oxide photocatalyst Z2S has better photocatalytic activity to bacteria inactivation than ZS, ZS2, Z and S films. Furthermore, nanostructured films were active even in the absence of irradiation.     

Synergistic inactivation of bacteria based on a combination of low frequency,low-intensity ultrasound and a food grade antioxidant

This study evaluated a synergistic antimicrobial treatment using a combination of low frequency and a low-intensity ultrasound (LFU) and a food-grade antioxidant, propyl gallate (PG), against a model gram-positive (Listeria innocua) and the gram-negative bacteria (Escherichia coli O157:H7). Bacterial inactivation kinetic measurements were complemented by characterization of biophysical changes in liposomes, changes in bacterial membrane permeability, morphological changes in bacterial cells, and intracellular oxidative stress upon treatment with PG, LFU, and a combination of PG + LFU. Combination of PG + LFU significantly (>4 log CFU/mL, P < 0.05) enhanced the inactivation of both L. innocua and E. coli O157:H7 compared to PG or LFU treatment. As expected, L. innocua had a significantly higher resistance to inactivation compared to E. coli using a combination of PG + LFU. Biophysical measurements in liposomes, bacterial permeability measurements, and scanning electron microscope (SEM)-based morphological measurements show rapid interactions of PG with membranes. Upon extended treatment of cells with PG + LFU, a significant increase in membrane damage was observed compared to PG or LFU alone. A lack of change in the intracellular thiol content following the combined treatment and limited effectiveness of exogenously added antioxidants in attenuating the synergistic antimicrobial action demonstrated that oxidative stress was not a leading mechanism responsible for the synergistic inactivation by PG + LFU. Overall, the study illustrates synergistic inactivation of bacteria using a combination of PG + LFU based on enhanced membrane damage and its potential for applications in the food and environmental systems.     

Preparation, characterization, and photocatalytic properties of silver carbonate

Silver carbonate (Ag₂CO₃) short rods were prepared using a precipitation method. It was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), diffuse reflective spectra (DRS) and photocatalytic degradation of organic pollutants and destruction of E. coli measurements. The results of DRS suggested that the optical transition of Ag₂CO₃ was indirectly allowed, and its band gap was determined to be 2.08 eV. The prepared Ag₂CO₃ displayed a high activity towards degradation of phenol and MB under visible light. The total organic carbon (TOC) value decreased during the photocatalytic process, which suggests that phenol was truly photodegraded. The stability of Ag₂CO₃ was greatly improved when Na₂CO₃ was added into the photocatalytic system. In addition, Ag₂CO₃ displayed enhanced photocatalytic activities for the destruction of E. coli due to its photogenerated electron-hole pairs.     

Analysis of sterilization effect by pulsed dielectric barrier discharge

Atmospheric pressure (AP) plasmas can sterilize against almost all kinds of bacteria because many ions and reactive species, such as oxygen atoms and ozone, etc., are generated during AP plasmas. So AP plasmas are proper processes for application to air cleaners and sterilizers. The aim of this paper is to evaluate a germicidal effect caused by pulsed plasma system in air utilizing a dielectric barrier discharge (DBD) type reactor incorporating alumina, glass, etc. Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa bacteria were used for this sterilization experiment. For analysis of the relationship between sterilization results and chemical species generated in the discharge, we used optical emission spectroscopy and we checked emission spectra by atomic oxygen (394.2 and 436.8 nm) and second positive system of nitrogen (337.1 nm). Experimental results showed that DBD treatment during 70 s sterilized E. coli with 99.99% effectively and ozone molecules were the dominant germicidal species. From these results we concluded that the pulsed DBD system is very effective for sterilization.     

Investigation of SiO2/Si3N4 films prepared on sapphire by r.f. magnetron reactive sputtering

Sapphire is a desired material for infrared-transmitting windows and domes because of its excellent optical and mechanical properties. However, its thermal shock resistance is limited by loss of compressive strength along the c-axis of the crystal with increasing temperature. In this paper, double layer films of SiO₂/Si₃N₄ were prepared on sapphire (α-Al₂O₃) by radio frequency magnetron reactive sputtering in order to increase both transmission and high temperature mechanical performance of infrared windows of sapphire. Composition and structure of each layer of the films were analyzed by X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD), respectively. Surface morphology and roughness of coated and uncoated sapphire have been measured using a talysurf. Flexural strengths of sapphire sample uncoated and coated with SiO₂/Si₃N₄ have been studied by 3-point bending tests at different temperatures. The results show that SiO₂/Si₃N₄ films can improve the surface morphology and reduce the surface roughness of sapphire substrate. In addition, the designed SiO₂/Si₃N₄ films can increase the transmission of sapphire in mid-wave infrared and strengthen sapphire at high temperatures. Results for 3-point bending tests indicated that the SiO₂/Si₃N₄ films increased the flexural strength of c-axis sapphire by a factor of about 1.4 at 800 °C.     

Mechanism and optimization of non-thermal plasma-activated water for bacterial inactivation by underwater plasma jet and delivery of reactive species underwater by cylindrical DBD plasma

Plasma-activated water (PAW) has been in use for the past decade in sanitization against bacteria and other microorganisms. This research study compared PAW generated by a DC positive flyback transformer (FBT) underwater plasma jet with delivery of reactive species underwater by cylindrical dielectric barrier discharge (C-DBD) with a neon transformer. A Box–Behnken design was adopted as a response surface methodology (RSM) to design the experimental plan and optimize operating parameters including time, gas flow, and gas ratio. The physical responses comprise optical emission spectroscopy (OES), pH, oxidation-reduction potential (ORP), and electrical conductivity (EC). The chemical responses consist of hydrogen peroxide (H₂O₂) and hydroxyl radicals (OH·). The biological responses include Escherichia coli reduction and Staphylococcus aureus reduction. The optimal condition for underwater plasma jet was found to be Ar gas with a flow rate of 3 slm for 6.5 min of treatment time, which can reduce E. coli and S. aureus to 7.14 ± 0.14 and 3.10 ± 0.26 in log, respectively. Also, the optimal condition for delivery of reactive species underwater by C-DBD plasma was found to be Ar (99%): O₂ (1%) gas mixture with an Ar gas flow rate of 4 slm for a treatment time of 11.5 min, which could reduce E. coli and S. aureus to 0.45 ± 0.07 and 2.45 ± 0.23 in log, respectively.     

Ultrasound-induced membrane lipid peroxidation and cell damage of Escherichia coli in the presence of non-woven TiO2 fabrics

A non-woven titanium dioxide (TiO₂) fabric was applied to disinfection by ultrasound (US) irradiation, and the disinfection efficiency and lipid peroxidation of Escherichia coli (E. coli) cell membrane were evaluated to investigate the killing process. The addition of non-woven TiO₂ fabric enhanced hydroxyl (OH) radical generation and disinfection efficiency. Judging from the disinfection experiments using glutathione or t-butanol as a radical scavenger, the OH radical played a major role in cell killing in sonodynamic disinfection with non-woven TiO₂ fabric. Moreover, to understand the detailed killing process, damage to cell membrane was also evaluated using a diphenyl-1-pyrenylphosphine (DPPP) fluorescent probe, which detects the membrane’s lipid peroxidation. The addition of non-woven TiO₂ fabric aggravated this peroxidation. This aggravation was caused by the OH radical according to an assay using a radical scavenger. From these results, it was concluded that non-woven TiO₂ fabric as a sonocatalyst promoted peroxidation of the polyunsaturated phospholipid component of the lipid membrane initially and induced a major disorder in the E. coli cell membrane under US irradiation.     

Ultrasonic-assisted synthesis of lignin-capped Cu2O nanocomposite with antibiofilm properties

Under ultrasonication, cuprous oxide (Cu₂O) microparticles (<5 µm) were fragmented into nanoparticles (NPs, ranging from 10 to 30 nm in diameter), and interacted strongly with alkali lignin (Mw = 10 kDa) to form a nanocomposite. The ultrasonic wave generates strong binding interaction between lignin and Cu₂O. The L-Cu nanocomposite exhibited synergistic effects with enhanced antibiofilm activities against E. coli, multidrug-resistant (MDR) E. coli, S. aureus (SA), methicillin-resistant SA, and P. aeruginosa (PA). The lignin-Cu₂O (L-Cu) nanocomposite also imparted notable eradication of such bacterial biofilms. Experimental evidence unraveled the destruction of bacterial cell walls by L-Cu, which interacted strongly with the bacterial membrane. After exposure to L-Cu, the bacterial cells lost the integrated structural morphology. The estimated MIC for biofilm inhibition for the five tested pathogens was 1 mg/mL L-Cu (92 % lignin and 8 % Cu₂ONPs, w/w %). The MIC for bacterial eradication was noticeably lower; 0.3 mg/mL (87 % lignin + 13 % Cu₂ONPs, w/w %) for PA and SA, whereas this value was appreciably higher for MDR E. coli (0.56 mg/mL, 86 % lignin and 14 % Cu₂O NPs). Such results highlighted the potential of L-Cu as an alternative to neutralize MDR pathogens.     

Microbial inactivation in cloudy apple juice by multi-frequency Dynashock power ultrasound

The study determined the efficacy of Dynashock wave power ultrasound as an alternative processing technique for apple juice against a number of pathogenic and spoilage microorganisms. The effects of several implicit, intrinsic and extrinsic properties on the Dynashock wave inactivation of the microorganisms were also investigated. Results showed that acid adaptation increased the resistance of Escherichia coli O157:H7 and Salmonella spp. but decreased that of Listeria monocytogenes. Spoilage yeast mixed inoculum composed of Debaryomyces hansenii, Torulaspora delbrueckii, Clavispora lusitaniae, Pichia fermentans and Saccharomyces cerevisiae was found to be more resistant than any of the adapted or non-adapted pathogens. Among the individual, acid-adapted E. coli O157:H7, the MN-28 isolate was found most resistant; while three other individual isolates had greater resistance than the composited E. coli inoculum. Increased in pulp content decreased the efficacy of Dynashock waves, but co-treatment with ultraviolet-C rays significantly enhanced inactivation in the cloudy apple juice. The results demonstrated the potential of Dynashock wave technology, together with other antimicrobial hurdles as alternative juice processing technique/s.     

Effect of supercritical CO2 on extremophile cells

The kinetics of inactivation of E. coli and D. radiodurance bacteria and Candida albicans imperfect fungi in compressed gaseous and supercritical carbon dioxide was studied. The possibility of decreasing the concentration of survived cells by 5 to 7 orders of magnitude was demonstrated.     

Asymptotic Expansions for λ d of the Dimer and Monomer-Dimer Problems

In the past few years we have derived asymptotic expansions for λ _d of the dimer problem and λ _d (p) of the monomer-dimer problem. The many expansions so far computed are collected herein. We shine a light on results in two dimensions inspired by the work of M.E. Fisher. Much of the work reported here was joint with Shmuel Friedland.     

Nondestructive monitoring,kinetics and antimicrobial properties of ultrasound technology applied for surface decontamination of bacterial foodborne pathogen in pork

In this study, electronic nose (E-nose) and Hyperspectral Imaging (HSI) was employed for nondestructive monitoring of ultrasound efficiency (20KHZ) in the inactivation of Salmonella Typhimurium, and Escherichia coli in inoculated pork samples treated for 10, 20 and 30 min.Weibull, and Log-linear model fitted well (R² ≥ 0.9) for both Salmonella Typhimurium, and Escherichia coli inactivation kinetics. The study also revealed that ultrasound has antimicrobial effects on the pathogens. For qualitative analysis, unsupervised (PCA) and supervised (LDA) chemometric algorithms were applied. PCA was used for successful sample clustering and LDA approach was used to construct statistical models for the classification of ultrasound treated and untreated samples. LDA showed classification accuracies of 99.26%,99.63%,99.70%, 99.43% for E-nose - S. Typhimurium, E-nose -E. coli, HSI - S. Typhimurium and HSI -E. coli respectively. PLSR quantitative models showed robust models for S. Typhimurium- (E-nose Rp² = 0.9375, RMSEP = 0.2107 log CFU/g and RPD = 9.7240 and (HSI Rp² = 0.9687 RMSEP = 0.1985 log CFU/g and RPD = 10.3217) and E. coli -(E-nose -Rp² = 0.9531, RMSEP = 0.2057 log CFU/g and RPD = 9.9604) and (HIS- Rp² = 0.9687, RMSEP = 0.2014 log CFU/g and RPD = 10.1731).This novel study shows the overall effectiveness of applying E-nose and HSI for in-situ and nondestructive detection, discrimination and quantification of bacterial foodborne pathogens during the application of food processing technologies like ultrasound for pathogen inactivation.     

Kinetic modeling of inactivation of natural microbiota and Escherichia coli on cherry tomato treated with fixed multi-frequency sonication

The suitability of some non-linear kinetic models (Weibull {with or without tail}, Log-linear, Log-linear shoulder {with or without tail}, Biphasic linear, Logistic, Multi-target and Single-target models) were evaluated to determine the inactivation kinetics of inoculated E. coli, and natural microbiota (i.e. mesophilic aerobic bacteria, and mold and yeast) on cherry tomato treated with fixed multi-frequency ultrasound. Almost all the studied model fitted well (R² ≥ 0.9) for the inactivation kinetics; however, the Weibull, Log-linear shoulder, and Biphasic linear model showed the highest statistical parameters (0.9 ≤ adj. R² ≤ 0.99 and smallest RMSE and SSE values). All the three models could be used to compare the kinetic behavior of E. coli and the first two models for the kinetic behavior of mesophilic aerobic bacteria and mold and yeast during sonication treatment. Two distinctive inactivation curves were obtained for the mono-frequency and the multi-frequency (dual and tri-frequency) for all the microbial inactivation. The remarkable results obtained for dual and tri-frequency sonication shows to be an effective and promising alternative to the traditional microbial inactivation techniques and the common practice of using ultrasound with other sanitizing methods.