Vital-Fluorochromization of Microorganisms Using 3′,6′-Diacetylfluorescein to Determine Damages of Cell Membranes and Loss of Metabolic Activity by Ozonation

Cell suspensions of Escherichia coli as a model for bacterial populations in wastewaters were treated with ozone as a disinfectant in a continuously operated pilot plant with a plug flow reactor. Suspensions with an initial number of CFU (colony forming units) of 10⁸ mL^?1 were ozonized with ozone concentrations up to 12 mg/L. Metabolic activities and membrane functions break down with increasing ozone concentrations. The fluorochromization using 3′,6′-diacetylfluorescein (FDA) proved to be a suitable method for the detection of an alteration in permeability of the cell membranes and an inactivation of metabolic activity. By fluorescence microscopic and photometric investigations it could be clearly demonstrated that, in the case of disinfection with ozone, inactivation of the cells includes first of all a damage of the cell membranes. In contrast to the determination of the number of CFU, fluorochromization allows the detection of alteration in metabolic activities.     

Inactivation of Giardia muris Using Ozone and Ozone-Hydrogen Peroxide

The disinfection effects of the ozone molecule alone and that of ozone decomposition products when inactivating Giardia muris cysts were investigated at bench-scale using two different ozone demand-free laboratory buffer systems. The first water was a 0.05 M phosphate buffer with hydrogen peroxide added at a 10:1 weight ratio. The second water was a 0.05 M phosphate – 0.01 M bicarbonate buffer which quickly scavenged radical species from ozone decomposition. The C3H/HeN mouse model was used to assess the infectivity of ozone treated cysts.

The phosphate-bicarbonate buffer system had significantly greater (P ≤ 0.05) inactivation of G. muris cysts than that observed in the phosphate buffer – peroxide system where ozone was completely decomposed in less than 120 s. Consequently, the design of ozone disinfection processes should maintain ozone residual for disinfection prior to the addition of hydrogen peroxide for the oxidation of other compounds.     

Ozone in the Laundry Industry—Practical Experiences in the United Kingdom

Since the early 1990s, the use of ozone in many commercial and industrial laundering applications has been evolving rapidly. Ozone allows washing to be conducted using cold water, thereby saving considerable heat energy and water consumption. Additionally, ozone enhances the wash process, resulting in a significant reduction in detergent dosage and number of rinses, thus saving water. Ozone/cold water cycles are gentler to fabrics, thus extending linen life. Finally, ozone/cold water laundering is beneficial for effluents, resulting in reductions in COD (chemical oxygen demand). Microorganisms are destroyed effectively in ozone-wash waters, and washing and drying cycles are shorter, thus saving labor. In this paper, the authors describe some specific case studies at commercial laundering installations in the UK, whereby the users of ozone have reaped major benefits, including enhanced microorganism kills/inactivation and significant cost savings.     

Seawater Ozonation of Bacillus subtilis Spores: Implications for the Use of Ozone in Ballast Water Treatment

The potential of ozone for disinfection of ships’ ballast water was investigated using Bacillus subtilis spores as an indicator. The effects of pH, presence of iron, and bacterial strain on disinfection efficacy in seawater, under simulated ballast conditions, were investigated. Ozone dosages of 9 mg/L (pH 7) and 14 mg/L (pH 8.2) and 24 h contact achieved a 4-log inactivation with the various oxidant residuals formed. Iron surface at a ratio to water of 9 m²/m³ impaired the oxidant residuals and the disinfection of spores. Different strains of B. subtilis resulted in different CT values. Ozone does not seem to be a good choice for the control of spore-forming organisms in ballast water, but may be suitable for the control of other species.     

Inactivation of Fusarium oxysporum Conidia in Soil with Gaseous Ozone – Preliminary Studies

ABSTRACT

In this study, the efficiency of gaseous ozone (O₃) injected in the soil as an oxidizing agent for the inactivation of F. oxysporum was evaluated under laboratory conditions. The results show the treatment reached an inactivation efficiency of 76% after an applied dose of 0.40 g O₃ kg ^?1 soil. This shows that the injection of O₃ can be a viable alternative to control pathogenic organisms in the soils. Nevertheless, it is clear that more studies on determining the effects of this treatment on soil quality are needed.     

Inactivation kinetics for Salmonella Enteritidis in potato omelet using microwave heating treatments

The aim of this study was to model the inactivation of Salmonella enterica serovar Enteritidis in pasteurized omelet internally inoculated at different microwave heating treatments (300 W; 450 W, 600 W and 800 W). Results indicated a non significant change in Salmonella populations during the first 30 s treatment at 300 W and 450 W, being log reductions lower than 0.5 log CFU g⁻¹. However, after 40 s treatment, log reductions had risen to 4.8 log CFU g⁻¹ at 800 W. Inactivation rates were higher at 600 W and 800 W (0.67 and 0.63 s⁻¹) than at 300 W and 450 W (<0.34 s⁻¹). The temperature-dependent parameters of a Weibull model obtained by Mattick, Legan, Humphrey & Peleg (2001) were evaluated. It was concluded that combinations characterized by a temperature equal or above 70 °C ensured a minimum 4 log reduction of Salmonella population (i.e. 300 W-80 s; 450 W-60 s or 600 W/800 W-40 s). These results may be of value in food service establishments, as target treatments for microwavable potato omelet portions.     

Inactivation efficacy of plasma-activated water: influence of plasma treatment time,exposure time and bacterial species

This study aimed to evaluate the influence of plasma treatment time, bacterial exposure time to PAW and bacterial species on the inactivation efficacy of plasma-activated water (PAW), with additional investigation of the inactivation mechanisms of PAW. Six bacterial species, including Listeria innocua, Staphyloccus aureus, Escherichia coli, Pseudomonas fluorescens, Shewanella putrefaciens and Aeromonas hydrophila were selected as the representative bacteria. The initial bacterial concentration was around 7 log CFU ml⁻¹ after mixing with PAW, and the inactivation efficacy was measured after different exposure times during the 4 °C storage. Scanning electron microscopy (SEM) images of the bacteria after PAW treatment were carried out to inspect the cell structure damage, and physicochemical properties of PAW, including pH, conductivity and long-living reactive species of H₂O₂, , and , were examined. The results showed that the inactivation efficacy of PAW was positively correlated with plasma treatment time and bacterial exposure time, and for the species examined in this study, the Gram-negative species were more sensitive to PAW than the Gram-positive species. Cell structure damage, including shrinkage, distortion, or holes, was observed after PAW treatment. The pH of PAW was acidified to 2.5–2.9, and conductivity was significantly increased to 518.0 μs cm⁻¹. and H₂O₂ were reduced during the 48 h storage, while an increased concentration was observed for . This study demonstrated that the processing parameters of plasma treatment time, exposure time and characteristics of bacteria can significantly affect the inactivation efficacy of PAW.     

Modelling of the inactivation kinetics of Escherichia coli,Saccharomyces cerevisiae and pectin methylesterase in orange juice treated with ultrasonic-assisted supercritical carbon dioxide

The combined effect of supercritical carbon dioxide (SC-CO₂) and high power ultrasound (HPU) on the inactivation kinetics of Escherichia coli, Saccharomyces cerevisiae and pectin-methyl esterase (PME) in orange juice was studied in order to select models that can predict their inactivation behaviour based on process parameters. Experiments were performed at different temperatures (31–41 °C, 225 bar) and pressures (100–350 bar, 36 °C). The inactivation rate of E. coli, S. cerevisiae and PME increased with pressure and temperature during SC-CO₂ + HPU treatments. The SC-CO₂ + HPU inactivation kinetics of E. coli, S. cerevisiae and PME were represented by models that included temperature, pressure and treatment time as variables, based on the Biphasic, the Peleg Type B, and the fractional models, respectively. The HPU-assisted SC-CO₂ batch system permits the use of mild process conditions and treatment times that can be even shorter than those of continuous SC-CO₂ systems.     

Inactivation of Enterobacter sakazakii by pulsed electric field in buffered peptone water and infant formula milk

The effect of high-intensity pulsed electric field (PEF) treatment on the survival of Enterobacter sakazakii suspended in buffered peptone water (BPW) and powdered infant formula milk (IFM) was evaluated. Reference medium and IFM samples were treated with PEF. Electric field intensity and treatment time were varied from 10 to 40 kV cm⁻¹ and from 60 to 3895 μs, respectively. Samples of buffered peptone water (3 g L⁻¹) and IFM were inoculated with E. sakazakii (CECT 858) (10⁹ cfu mL⁻¹) and then treated with PEF. The inactivation data were adjusted to the Weibull frequency distribution function and Bigelow model, and constants were calculated for both substrates. A maximum 2.7 log (cfu mL⁻¹) reduction was achieved in BPW after exposure of E. sakazakii to PEF for 360 μs (2.5 μs pulse width) at 40 kV cm⁻¹. In IFM, exposure of E. sakazakii to PEF, with the same conditions, led to a 1.2 log (cfu mL⁻¹) reduction. The greater the field strength and treatment time, the greater the inactivation achieved in both substrates. Even though further research will be necessary, according to the results, there are good prospects for the use of PEF in hospitals to achieve safe reconstituted infant formula before storage at refrigerated temperatures.     

Inactivation of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri on spinach leaves by X-ray

Several recent foodborne disease outbreaks associated with leafy green vegetables, including spinach, have been reported. X-ray is a non-thermal technology that has shown promise for reducing pathogenic and spoilage bacteria on spinach leaves. Inactivation of inoculated Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri on spinach leaves using X-ray at different doses (0.1, 0.2, 0.3, 0.5, 0.75, 1.0, 1.5 and 2.0 kGy) was studied. The effect of X-ray on color quality and microflora counts (mesophilic counts, psychrotrophic counts and yeast and mold counts) of untreated and treated spinach was also determined. A mixture of three strains of each tested organism was spot inoculated (100 μl) onto the surface of spinach leaves (approximately 8–9 log ml⁻¹), separately, and air-dried, followed by treatment with X-ray at 22 °C and 55–60% relative humidity. Surviving bacterial populations on spinach leaves were evaluated using a nonselective medium (tryptic soy agar) with a selective medium overlay for each bacteria; E. coli O157:H7 (CT-SMAC agar), L. monocytogenes (MOA), and S. enterica and S. flexneri (XLD). More than a 5 log CFU reduction/leaf was achieved with 2.0 kGy X-ray for all tested pathogens. Furthermore, treatment with X-ray significantly reduced the initial inherent microflora on spinach leaves and inherent levels were significantly (p < 0.05) lower than the control sample throughout refrigerated storage for 30 days. Treatment with X-ray did not significantly affect the color of spinach leaves, even when the maximum dose (2.0 kGy) was used.